Oral Abstracts

SESSION PLENI ERE: METABOLISMES, DU GENE AU group). Neural activity of the centers is studied through the low frequency of amplitude oscillations (LFAO) of fMRI signal of the selected brainstem regions of PATIENT interest. It is a validated index of fMRI neural activation. Results: COPD patients display a higher level of airway ﬂow complexity as com- Contribution of bone to whole-body metabolic regulation pared with healthy subjects. In controls and COPD patients ventilatory complex- G Karsentya, PA Marksa aDepartment of Genetics & Development, Columbia ity correlates with the activity of the respiratory neural centers. During spontaneous breathing in controls, inspiratory chaos is tightly linked with the University Medical Center, 701 W 168th Street, Room 1602A HHSC, New York, 2

gy = = NY, USA LFAO of the ventro-lateral (VL) medulla (R 0.75, P 0.01) while in COPD patients, expiratory chaos correlates with LFAO of the VL pons (R2 = 0.4, Bone is the only tissue of the vertebrate body that contains a cell type, the osteo- = clast, whose only function is to destroy the host tissue. This does not occur at P 0.03). External inspiratory resistive overload inhibits brainstem fMRI signal lo random but rather in the context of a well ordered physiological process called while significantly decreasing the resulting airway flow complexity in parallel in bone modeling during childhood and remodeling during adulthood. We hypothe- both populations. o sized that this process requires a daily delivery of a large amount of energy to Conclusions: We present the first study showing that airway flow complexity is bone cells; a large body of clinical observations supports this hypothesis. Testing strongly depending on the activity of the respiratory central pattern generators c this hypothesis led us to reveal the bone endocrine nature of bone and to identify assess with fMRI. COPD patients have a high neural ventilatory drive due to osteocalcin as an osteoblast-derived hormone affecting multiple aspects of energy chronically enhance respiratory load. We show in patients, at rest and during an a metabolism. We will review during this lecture the functions and mode of action increase load, a high level of airway flow complexity that translates the altered of osteocalcin and address its overall importance in the control of whole body high activity of the brainstem respiratory centers. These findings may be involved glucose homeostasis. in the onset of respiratory failure when the neural network becomes inefficient to sustain respiratory overload. m References: r *Mangin L., et al. Res. Physiol. Neurobiol. (2008) 161(2) 189–196. Mangin L., et al. Am. J. Physiol. (2009) 296(4) R1088–R1097. a Futur developments in personalized human obesity management: is Mangin L., et al. PLoS One (2011) 6(1) e16297. systemic medicine usefull? Fundings: PHRC, BQR Paris 7, Fond de dotation Recherche Respiratoire a a Keywords: respiratory neural network-chaos-breathing-fMRI-COPD. h K Clement Institut Cardiometabolisme et Nutrition (ICAN), INSERM/UPMC, Hôpital de la Pitie-Salpêtriere, Paris

P Abstract not available: 01-02

Optoelectronic plethysmography in suspected and diagnosed unilateral diaphragm weakness J Boudarhama, D Pradona, H Prigentb, L Falaizeb, MC Durandb, H Mericb, SESSION 1: GS MIXTE RESPIRATION F Lofasob, M Petitjeanc aPlateforme d’Analyse du Mouvement et Centre d’Innovations Technologiques UMR 805, Hôpital Raymond Poincare, Garches, France; bPhysiologie- Bitter taste receptors in the lung: a new pharmacological target? Explorations Fonctionnelles, Hôpital Raymond Poincare, EA 4497 Universitede S Grassin-Delylea aUPRES EA220, Hôpital Foch, Universite Versailles Saint Quentin Versailles Saint-Quentin-en-Yvelines, Garches, France; cCentre Hospitalier Universitaire en Yvelines, Saint Quentin en Yvelines d’Amiens-Picardie, EA 4497 Universite de Versailles Saint-Quentin-en-Yvelines, Bitter taste receptors (TAS2Rs) are known for long for their role in taste as sen- Amiens, France sors of the presence of toxic compounds in foods, but their unexpected expression Objective: The objective was to determine whether optoelectronic plethysmogra- in airways epithelium and smooth muscle cells or in peripheral blood leucocytes phy (OEP) can detect asymmetrical ventilation related to unilateral or asymmetri- Clinical has been recently documented. This family of GPCRs includes about 25 members cal diaphragmatic weakness, suggesting usefulness as a diagnostic tool. in humans and each subtype has a variable selectivity towards bitter compounds, Material and methods: Thirteen patients with suspected asymmetrical dia- some of them being restrictedly selective to a unique molecule and others phragmatic dysfunction based on dyspnea and hemidiaphragm elevation on the responding to a wider range. More than a hundred molecules such as chloro- chest radiograph were studied, as well as three patients with maltase acid defi- quine, caffeine, strychnine, colchicine or erythromycin have thus been described ciency (a cause of symmetrical diaphragmatic weakness). The transdiaphragmatic as TAS2R agonists while TAS2R19, 41, 42, 45 and 60 are considered as pressure response to unilateral magnetic stimulation (latPdiTw) and the dia- orphans since no agonist has been identified. phragm compound muscle action potentials (CMAPs) elicited by transcutaneous In the airways, the initial observation by Desphandes et al. (2010) described the electrical stimulation of each phrenic nerve. Chest wall kinematics was studied in ntal & relaxation of pre-contracted mouse trachea following exposure to chloroquine, the supine position using a motion analysis system (Motion Analysisâ, Santa e denatonium, quinine or saccharine, which was suggested to be even more pro- Rosa, USA) during spirometric tidal volume (VT) and inspiratory capacity (IC) nounced that the relaxation obtained with the reference relaxing agents b2- measurement with a Vmax 229 Sensormedics System (Yorba Linda, USA). Fifty- adrenoreceptor agonists. Interestingly, an original intracellular signaling pathway two reflective markers were placed over the anterior chest wall from the clavicles in the response of airway smooth muscle cells to bitter-taste receptor agonists to the pubic bone. To compute volumes, chest wall surface area was calculated was proposed, involving the G-protein bc subunit and leading to a localized from a triangular mesh created by connecting the nodes represented by the increase in intracellular calcium, which in turn causes membrane hyperpolarisa- reflective markers, and we used standard algorithms to obtain the total chest cav- tion through an activation of large conductance potassium channels (BK Ca). In ity volume and volume changes in each thoracoabdominal compartment. Then, addition to these results in cell cultures or airways preparations, inhaled bitter ta- each compartment was separated into two sides (affected side and other side). stants were shown effective in decreasing airway resistance in ovalbumin-sensi- Because in 12 healthy controls, we found that the inspired volume measured on tized mice, but very little is known in humans to date. However, transcriptome one side during VT or IC measurement was never >55% or smaller than 45% of analysis revealed upregulation of TAS2R signaling in peripheral blood leucocytes the total inspired volume values above or below these limits were taken to indi-

Fundam from patients with severe asthma, as well as a correlation between clinical mark- cate unilateral or asymmetrical diaphragm dysfunction. ers of asthma severity and TAS2R expression. Results: The CMAPs and latPdiTw showed unilateral or predominantly unilateral Overall, these works suggest that bitter taste receptors may constitute a new diaphragmatic dysfunction in nine of the 13 patients. By OEP, the affected side pharmacological target for obstructive lung diseases such as asthma and COPD. contributed <45% of the inspiratory capacity in each of these nine patients, We will address the role of bitter taste receptors in respiratory pharmacology, whereas no asymmetry was noted in the other four patients or in the three with a special focus on results obtained in human tissues. patients with maltase acid deficiency. All patients preferred OEP over CMAP or latPdiTw. Discussion: OEP detected asymmetric ventilation in all patients diagnosed with 01-01 diaphragm weakness and in no patients without this diagnosis. Thus, OEP is an Neural substrates of ventilatory chaos in humans effective noninvasive alternative that is preferred by the patients over CMAP A Hessa,LYub,c, I Kleind, M de Mazancourte, G Jebrakf, H Malf, M Fournierf, response and latPdiTw. E Schouman-Clayesd, M Courbageg, C Clericih, L Mangini aLaboratoire Matiere et Keywords: plethysmography; chest wall; diaphragm; phrenic nerve. Systemes Complexes CNRS UMR7057 Universite Paris 7, Service de Radiologie Hôpital Bichat, Paris, France; bLaboratoire Matiere et Systemes Complexes Universite Paris 7, Paris, France; cInstitute of Theoretical Physics, Lanzhou University, Lanzhou; 01-03 dService de Radiologie Hôpital Bichat, Paris; eEcole Normale Superieure, Laboratoire NGF contributes to medial and intimal remodelling of pulmonary Matiere et Systemes Complexes Universite Paris 7, Paris, France; fService de arteries in experimental pulmonary hypertension: receptors and Pneumologie Hôpital Bichat, Paris, France; gLaboratoire Matiere et Systemes signalling pathways involved Complexes CNRS UMR7057 Universite Paris 7, Paris, France; hInserm 700 V Freund-Michela, F Costeb, M Duboisa, A Courtoisa, R Marthanb, JP Savineaua, Universite Paris 7, Service de Physiologie Hôpital Bichat, Paris, France; iLaboratoire B Mullera aUniversite Bordeaux Segalen, INSERM U1045, Bordeaux, France; Matiere et Systemes Complexes CNRS UMR 7057 Universite Paris 7, Service de bUniversite Bordeaux Segalen, INSERM U1045, CHU de Bordeaux, Bordeaux, France Physiologie Hôpital Bichat, Paris, France Introduction: We have previously shown that expression of the nerve growth Objectives: Biological systems exhibit nonlinear deterministic dynamics that can factor NGF is increased in pulmonary hypertension (PH), with NGF playing a role be chaotic, either at a cellular, organ or system levels. Human ventilation exhib- in various PH pathophysiological aspects. We have here studied whether NGF is its chaotic behavior, as well*. Direct evidences that ventilatory complexity is involved in medial and intimal remodelling of pulmonary arteries in vivo in PH linked to the activity of the respiratory neural centers are still lacking in rat models. Receptors and signalling pathways involved in NGF-induced prolifera- humans. tion of human pulmonary arterial smooth muscle (PASMC) and endothelial cells Methods: In 25 healthy subjects and 25 patients with chronic obstructive pul- (PAEC) in vitro were also investigated. monary disease (COPD), we evaluate airway flow chaos (noise titration, largest Methods: Experimental PH in the rat was induced either by a single injection of Lyapunov exponent and correlation dimension). Brainstem respiratory centers monocrotaline (MCT, day (D1, 60 mg/kg), or after 28 days of chronic hypoxia are located with cerebral functional magnetic resonance imaging (fMRI) in the (CH, 0.5 atm). Anti-NGF blocking antibodies (10 lg/kg ip) were administered as ventro-lateral (VL) medulla (PreBotzinger€ complex) and the VL pons (parafacial a preventive treatment at D0-2-7 for MCT or at D1-8-15-22 for CH. Antibodies

were also administered as a curative treatment at D15-19-22-26 for both MCT of 613 Æ 182 mL/min (+96%). Exercise was also associated with a decrease and CH. Pulmonary arterial pressure was assessed at D28. Pulmonary arterial (P<0.0146) in Rrs of 5.3 Æ 2.82 hPa s/L (À24%) authenticating a concomi- luminal occlusion, intimal or medial thickness were evaluated on lung sections tant bronchodilation. During exercise, the incidence of the cough reflex (CR) after hematoxylin and eosin stainings, or after immunohistochemical stainings decreased and the expiration reflex (ER) increased (P<0.0001). Whereas the with anti-von Willebrand factor or anti-a-smooth muscle actin antibodies. NGF- sensitivity of the CR and the ER are both decreased during exercise compared to induced proliferation of PASMC and PAEC in primary culture was assessed by the sensitivity of the CR at rest (P < 0.02), the strength of the expulsive response the BrdU technique. is unchanged. Results: In both MCT and CH rats, anti-NGF blocking antibodies displayed pre- Central findings of this study are that during exercise, the incidence of the cough ventive as well as curative effects on medial and intimal remodelling of pulmo- reflex is decreased and the cough threshold for evoking cough is increased, nary arteries. In vitro, NGF induced PASMC proliferation at low concentrations whereas the strength of cough is unchanged. These results indicate that adjust- through activation of the TrkA receptor and PI3-kinase dependent pathways. ments occurring during exercise likely down-regulates tracheal defensive reflexes NGF also induced PAEC proliferation, but only at higher concentrations through in anesthetized rabbits. activation of both TrkA and p75NTR receptors and PI3-kinase independent signal- Keywords: Cough, Expiration reflex, Mechanical stimulation, Exercise, Rabbit. ling pathways. Conclusion: We show here that anti-NGF blocking antibodies administered in vivo both prevent and reverse medial and intimal remodelling of pulmonary 01-06 arteries in PH rat models. NGF may contribute to pulmonary arterial remodelling Anatomical determinants of airway hyperresponsiveness: contribution of by stimulating proliferation of both PASMC and PAEC, but through activation of airway size assessed by anatomic dead space measurements distinct receptors and signalling pathways. These results suggest that blocking L Plantiera, C Delclauxb aAP-HP, Hôpital Bichat-Claude Bernard, Service de both NGF receptors may be of therapeutical interest in pulmonary arterial remod- Physiologie-Explorations fonctionnelles – Universite Paris Diderot, Paris, France; bAP- elling, and therefore confirms the interest of targeting NGF in pulmonary hyper- HP, Hôpital Europeen Georges Pompidou, Service de Physiologie-Explorations tension. fonctionnelles – Universite Paris Descartes, Paris, France Keywords: nerve growth factor NGF, pulmonary hypertension, pulmonary arte- Introduction: Contribution of anatomic factors to airway hyperresponsiveness rial remodelling. (AHR), a hallmark feature of asthma, has been suggested by correlations between surrogate markers of airway size relative to lung size (FEF25-75/FVC, diameter of main bronchi reported to lung area on chest radiographs) and quantitative mea- 01-04 surements of AHR (methacholine dose-response slope). The aim of this study is to Effect of oro-phayngeal povidone-iodine preventive oral care on directly determine whether subjects with AHR have lower airway volume ventilator-associated pneumonia in severe brain injury or cerebral reported to lung volume, in comparison with subjects without AHR. haemorrhage patients: a multicentre randomized controlled trial for the Methods: Ongoing prospective case-control study. Patients referred for explora- SPIRIT-ICU study and AtlanRea groups tion of chronic cough or intermittent dyspnea undergo baseline plethysmographic B Laviollea, P Seguinb, Y Malledantb, E Bellissanta aCHU de Rennes, Universitede lung function testing. Airway volume is quantified by capnographic measure- Rennes 1, CIC Inserm 0203, Rennes, France; bCHU de Rennes, Universite de Rennes ments of anatomical dead space using the Bohr and Fowler methods, before and 1, Inserm U991, Rennes, France after methacholine challenge up to a maximal cumulative dose of 1600 lg. AHR Objectives: Oral use of povidone-iodine to prevent ventilator-associated pneumo- is defined by FEV1 decline >15% compared to baseline. Patients with a history of nia remains unclear especially in high-risk patients [1, 2]. We assessed the effi- any chronic lung disease are not included. Preliminary data are presented as cacy and safety of oral care with povidone-iodine on the incidence of ventilator- means Æ SD. T-tests have been performed for informational purposes. associated pneumonia in a high-risk population. Results: Up to November 26th, 2012, n = 17 patients without AHR (Controls) Methods: This was a multicentre, placebo-controlled, randomised, double-blind, and n = 6 patients with AHR have been studied. Mean FEV1 (%) was 96 Æ 14 two-parallel-group trial performed in patients with severe (Glasgow coma in Controls and 86 Æ 14 in AHR patients (P = 0.07), while FEF25-75/FVC was scale 8) brain injury or cerebral haemorrhage requiring mechanical ventila- 0.8 Æ 0.24 in Controls and 0.68 Æ 0.18 in AHR patients (P = 0.14). Bohr and tion. Participants were assigned to receive oropharyngeal care with povidone- Fowler dead space indices were closely correlated (R2 = 0.61). Bohr dead space iodine (n = 91) or placebo (n = 88) six times daily until extubation. Primary en- reported to TLC was 3.6 Æ 0.16% in Controls and 2.6 Æ 0.65% in AHR patients point was the rate of ventilator-associated pneumonia. Secondary end point (P = 0.075); a trend towards lower Fowler dead space was likewise observed in included the rates of ventilator-associated tracheobronchitis and acute respiratory AHR patients (P = 0.085). Variation of both airway volume indices after meth- distress syndrome. acholine inhalation, a surrogate for smooth muscle contraction, was similar in Results: The number of patients evaluable for the primary endpoint (preplanned both groups (P = 0.36). modified intention-to-treat population) was 150 (78 in the povidone iodine Discussion: Pending confirmation, preliminary results suggest a possible strong group, and 72 in the placebo group). Ventilator-associated pneumonia occurred association between decreased direct measures of baseline airway volume and in 24 (31%) in the povidone-iodine group and 20 (28%) in the placebo group AHR, while surrogates of bronchial smooth muscle contraction do not appear (RR 1.11, 95% CI 0.67–1.82, P = 0.69). There was no significant difference increased in patients with AHR. between the two groups for ventilator-associated tracheobronchitis: eight patients Keywords: Airway, dead space. (10%) in the povidone-iodine group and 5 (7%) in the placebo group (RR 1.48, 95% CI 0.51–4.31, P = 0.47). Acute respiratory distress syndrome occurred in five patients in the povidone-iodine group and zero patient in the placebo group 01-07 (P = 0.06). Quadriceps and intercostal muscles oxygenation during maximal exercise Conclusion: There is no evidence to recommend oral care with povidone-iodine in altitude hypoxia: is there a perfusion competition between these to prevent ventilator-associated pneumonia in high-risk patients. Moreover, this muscles? strategy seems to increase the rate of acute respiratory distress syndrome. C De Bisschopa, S Belokab, H Groepenhoffc, M Van Der Plasd, M Overbeekc, References: R Naeijeb, H Guenarde aLaboratoire MObilite Vieillissement et Exercice (MOVE), 1. Seguin P.,Tanguy M., Laviolle B., et al. Effect of oropharyngeal decontamina- Poitiers, France; bDepartment of Pathophysiology, Faculty of Medicine, Bruxelles, tion by povidone-iodine on ventilator-associated pneumonia in patients with head Belgium; cDepartment of Pulmonology, VU University Medical Center, Amsterdam, trauma. Crit. Care Med. (2006) 34 1514–1519. The Netherlands; dDepartment of Respiratory Medicine, Onze Lieve Vrouwe Gasthuis, 2. Labeau S.O., Van de Vyver K., Brusselaers N., et al. Prevention of ventilator- Amsterdam, The Netherlands; eLaboratory of Physiology, Medical Faculty, Bordeaux, associated pneumonia with oral antiseptics: a systematic review and meta-analy- France sis. Lancet Infect. Dis. (2011) 11 845–854. Aim: At maximal exercise, the arterial oxygen flow to active muscles becomes Keywords: ventilator-associated pneumonia, head injury, oropharyngeal decon- insufficient to meet the oxygen demand. A competition between respiratory and tamination, povidone-iodine. peripheral muscles perfusions favouring the former might occur with a redistribu- tion of blood flow in favour of respiratory muscles. The purpose of the study was to investigate the relative adaptation of intercostals and thigh muscles during exercise 01-05 in the hypoxic condition of altitude in lowlanders as well as in highlanders. Due to Desensitization of cough during limb muscle contraction in anesthetized adaptation to hypoxia, differences between the groups of subjects were expected. rabbits Method: Eighteen recently acclimatized lowlanders and 27 highlanders (13 M Poussela, S Varechovaa, G Bosserb, B Demoulina, B Chalona, O Ruckebuscha, healthy subjects, and 14 with Chronic Mountain Sickness, (CMS)) were investi- A Tiotiua, C Schweitzera, B Chenuela aLaboratoire de Physiologie, EA 3450, gated in Cerro de Pasco (4350 m, Perou). The subjects underwent an incremen- Universite de Lorraine, Vandoeuvre-les-Nancy, France; bNutrition-Genetique et tal cardio-pulmonary exercise test on cycle ergometer. The exercise test was Exposition aux Risques Environnementaux, INSERM U 954, Universite de Lorraine, performed three times in lowlanders (sea-level, arriving at altitude and a week Vandoeuvre-les-Nancy, France later). During exercise, regional tissue oxygenation (rSO2) and tissue haemoglo- Cough can be evoked by mechanical or chemical irritation of the receptor field of bin concentration (Hbt) of intercostal muscles and vastus medialis were moni- the vagus nerve and expels inhaled foreign matter from the lungs or clears the tored by near-infrared spectroscopy (NIRS, Nonin Regional Oximetry Technology, airways of endogenous mucus. The primary role of this defensive reflex is to pro- Plymouth, MN, USA). tect the airways from potentially harmful agents and afferents located in the air- Results: Intercostal and vastus medialis-rSO2 were lower at altitude than at sea way mucosa are most relevant to this physiological function. The ‘cough center’ level (À8% to À12%) and decreased similarly during incremental exercise exhibits plasticity at the sensor and integration levels leading to modulation of (P < 0.05 from rest, above 60% VO2max). The time patterns of vastus medialis the reflex in its strength or in its pattern. Little is known about interactions and intercostal-rSO2 were similar in highlanders. At maximal exercise and during between cough and human activities, especially during exercise. The present recovery, intercostal-rSO2 was lower than vastus medialis-rSO2 in lowlanders. study was designed to determine whether exercise, mimicked by electrically- Conversely in highlanders, intercostal-rSO2 was greater than vastus medialis- induced muscular contractions, is able to change the occurrence and /or the rSO2, from rest to 80% VO2max. Changes in intercostal-rSO2 in highlanders were strength of cough following a mechanical stimulation of the trachea in anesthe- smaller than those in lowlanders at exercise. Intercostal and vastus medialis-Hbt tized rabbits. increased from sea level to altitude in lowlanders. [Hb] in lowlanders was Thirteen anesthetized, tracheotomized rabbits were studied allowing the analysis increased after a week in altitude but was lower than in CMS. of 47 sequences (including rest and exercise) and an overall of 311 tracheal Conclusion: Compared to sea level, maximal exercise in hypoxia did not alter mechanical stimulations: 196 at rest and 115 during exercise. Minute ventilation the kinetic of rSO2 and Hbt in peripheral muscle. At maximal exercise, even in was larger during exercise compared to baseline (P < 0.0001) with an increase acute hypoxia, the increase in ventilation did not alter lowlander’s peripheral

muscle oxygenation. Lower changes in intercostal-rSO2 were observed at maxi- 01-10 mal exercise in highlanders likely due to lower ventilation. Overall these findings New oral anticoagulants and oral pulmonary arterial hypertension do not favor the hypothesis of a blood shift to respiratory muscles. therapies: potential pharmacological interactions Keywords: Intercostal muscle, quadriceps, muscle oxygenation, NIRS, exercise, L Bertolettia, X Delavenneb, D Montanic,d, JC Legae,f, P Mismettig aGroupe de high altitude. Recherche sur la Thrombose, Service de Medecine et Therapeutique, CHU de St-Etienne, Universite Jean-Monnet, Saint-Etienne, France; bGroupe de Recherche sur la Thrombose, Laboratoire de Pharmacologie, CHU de St-Etienne, Universite Jean-Monnet, 01-08 Saint-Etienne, France; cAP-HP, Service de Pneumologie et Reanimation Respiratoire, ^ Which relationship between Hb conductance for CO and PO is the right Centre National de Reference de l’Hypertension Pulmonaire Severe, Kremlin-Bicetre, 2 d one in vivo in human? France; INSERM U999, Hypertension Arterielle Pulmonaire: Physiopathologie et a b c a ^ H Guenard , JB Martinot , C Kays Universite Bordeaux 2 et CHU, Bordeaux, Innovation Therapeutique, Centre Chirurgical Marie-Lannelongu, Kremlin-Bicetre, b c e ^ France; Clinique Ste Elisabeth, Namur, France; Universite et CHU de Bordeaux, France; Service de Medecine Interne et Vasculaire, Hopital Lyon Sud, Hospices Civils f Bordeaux, France de Lyon, Pierre-Benite, France; Groupe de Recherche sur la Thrombose, Universite g Aim: The aim was to select among the several published equations relating the Jean-Monnet, Pierre-Benite, France; CHU de St-Etienne; Universite Jean-Monnet conductance of Hb for CO (hCO) to PO2 the one(s) which would gave the same (EA 3065), Saint-Etienne, France DmCO/Vc ratios in the conditions of slight hypoxia (Pcap O2: 82 mmHg) and Background: Antithrombotic therapy (mainly with vitamin-K antagonisms: normoxia (118 mmHg). The assumption made was that the slight change in VKA) is currently recommended in idiopathic and familial Pulmonary Arterial PO2 would not change Vc and DmCO, as it would change TLCO. Hypertension (PAH) and in PAH associated with anorexigens. As VKA are cur- Method: Eight subjects were included. They performed duplicated measurements rently deeply challenged by new oral anticoagulants (NOA) in the setting of of CO and NO lung transfer using the single breath method (Hypercompact, Med- venous thrombo-embolism and atrial fibrillation, we searched for potential phar- isoft Dinant B) with either a slightly hypoxic mixture (15% O2) or the standard macological interactions between NOA and oral PAH therapies. mixture (19%) Standard concentrations of inspired NO (40 ppm) and CO Methods: We reviewed the potential pharmacokinetic and pharmacodynamics (2000 ppm) were used. All calibrations were made before and after a session drug-drug interactions (DDI), in particularly regarding metabolism and drug with a given subject. DmCO and Vc were calculated using the Roughton Forster transport, with two endothelin-receptor antagonisms (ERA): bosentan (B) and (1957) equation and a conductance of Hb for NO of 4.5 mL/(min mmHg) ambrisentan (A), two phosphodiesterase-5 inhibitors (PDE5i): sildenafil (S) and according to Borland et al (2010). tadalafil (T), and NOA (rivaroxaban, apixaban, dabigatran). Results: Dm/Vc ratios were calculated in slightly hypoxic and normoxic condi- Results: Within ERA, B is mainly metabolized by hepatic cytochrome P450 tions. Seven published equations cited by Hughes and Bates were tested. Only (CYP) 3A4, A by uridine 5’ diphosphate glucuronyltransferase and to a lesser three equations gave Dm/Vc ratios different from <10% [Holland 2.47 (hypoxic) extent, by CYP3A4 and CYP2C19. The organic anion transport proteins for B vs. 2.58 (normoxic), Forster 2.46 vs. 2.70, Roughton Forster (k = 1.5) 3.43 vs. and P-glycoprotein for both are probably involved in the transports of these 3.23]. This last equation was however not kept as deriving from in vitro experi- drugs. B, but not A, induces CYP3A4, which is involved in the metabolism of ments at pH 8. The two remaining equations gave similar results for Vc and anti-Xa NOA rivaroxaban (30%) and apixaban (50%). Concomitant use of B may DmCO. The differences for Vc were+2.4 and À1.7 mL for Holland and Forster diminish their biological efficacy. Regarding PDE5i, S and T are also mainly respectively as those for DmCO were +12 and +9 mL/(min mmHg). These differ- metabolized by CYP3A4, but act as slight CYP3A4 inhibitors. The risk for clini- ences were <10% of the raw values. Both equations are acceptable for practical cally significant DDI seems small between anti-Xa NOA and S or T. The effect of use of diffusion tests as all others would give inaccurate values. A compromise PAH-combination therapy is unknown but it may decrease deeper the concentra- between the the selected equations would be 1/hCO = 1.19 + 0.0053 PO2 tion of anti-Xa NOA. Conversely, dabigatran (an anti-IIa drug not metabolised by (mmHg). CYP) should be avoided in PAH because of its potential increased risk of myocar- Conclusion: The present results could mark the end of decades of uncertainty dial infarction. on Vc and DmCO values due to the scatter of the hCO values used. It is worthy Conclusion: Concomitant use of PAH therapies (mainly bosentan) and NOA to note that the hNO value used here leads to high DmCO values, close to mor- may expose patients to DDI. In the absence of robust clinical and pharmacologi- phometric data at least during exercise. cal data, NOA are not recommended in PAH. Keywords: lung diffusion, carbon monoxide, nitric oxide, blood conductance, Keywords: Bosentan, ambrisentan, sildenafil, tadalafil, rivaroxaban, apixaban, hypoxia. dabigatran, pulmonary arterial hypertension, drug-drug interaction.

01-09 01-11 Microparticles induce pulmonary artery contraction through activation Can we reduce use of third-generation cephalosporins and of acid sphingomyelinase and inhibition of Kv currents fluoroquinolones in lower respiratory tract infections in the emergency J Moral Sanza, B Barreraa, L Morenoa, D Morales Canob, R Soletic, F Grolleauc, department? c a c a a E Montassiera, TX Limb, N Goffinetc, P Le Contec, G Poteld, E Batarda aEA3826, R Andriantsitohaina , A Cogolludo , Mc Martinez , F Perez Vizcaino Universidad b c d Complutense de Madrid, Madrid, Spain; bUniversidad Complutense de Madrid, Angers, Nantes, France; Urgences, 44000 Nantes, France; Urgences, Nantes, France; EA France; cINSERM U1063, Angers, France 3826, Nantes, France Introduction: Microparticles are membrane vesicles from damaged or activated Objective: Fluoroquinolones and 3rd-generation cephalosporins (3GC) have cells that are associated with atherosclerosis, cancer and inflammatory diseases. become the antibiotics of choice in many hospitals in recent years for the treat- Long-term exposure to microparticles induces vascular hyporeactivity in aortae ment of infections such as a Lower Respiratory Tract Infection (LRTI). Because via the Fas/FasL pathway and endothelial dysfunction in pulmonary arteries fluoroquinolones and 3GC select multiple bacterial resistances, usage of both clas- (PA). We hypothesized that microparticles have acute effects on pulmonary vas- ses should be restricted. Our objectives were to assess the frequency of prescrip- cular contractility and Kv channel function. tion of 3GC and fluoroquinolones in LRTI and to evaluate the avoidable part of Methods: Resistance PA (300–500 lm internal diameter) were isolated from these prescriptions. male Wistar rats and PA myocytes were isolated by enzymatic digestion. Vascu- Methods: Retrospective serie of 88 adult cases of community-acquired pneumo- lar reactivity was assessed using isometric wire myographs. The responses nia and acute exacerbation of COPD admitted to the emergency department. induced by microparticles from apoptotic T lymphocytic cells (10 mg protein/mL) Compliance with French national guidelines was assessed. Furthermore, prescrip- or the FasLigand (10 mg/mL) were measured after 30 min. Some experiments tions that complied with guidelines were deemed justified if at least one criterion were performed in the presence of nifedipine (1 mM), tiron (1 mM), an anticera- was present: allergy or intolerance to penicillin, failure of penicillin, penicillin mide antibody (200 ng/mL) or inhibitors of the acid (D-609, 100 mM) or neutral therapy in the previous month, admission in ICU, and only for quinolones, sus- sphingomyelinases (GW4869, 10 mM). Potassium currents and membrane poten- pected legionellosis. Avoidable prescriptions included prescriptions that did not tial were recorded using the patch clamp technique in freshly isolated PASMC. comply with national guidelines and prescriptions that complied with guidelines Reactive oxygen species (ROS) were measured in human PASMC in culture incu- but were not justified. Avoidable prescriptions of 3GC and fluoroquinolones may bated with dichlorofluorescein. be replaced by amoxicillin/clavulanate. Results: Microparticles reduced Kv current amplitude (37 Æ 6% inhibition at Results: We included 72 pneumonia, 16 acute exacerbation of COPD. Median +30 mV; P < 0.01) and produced membrane depolarization in PASMC (from age was 79 (32–94). Thirty-two [36.4% (27.1–46.8)] and 10 [11.3% (5.2– À32 Æ 2toÀ20 Æ 4 mV; P < 0.05). Moreover, microparticle-induced a slow 20.1)] patients received respectively 3GC and fluoroquinolone. The majority of developing contraction in PA (8 Æ 2% of KCl contraction after 30 min) that was prescriptions of fluoroquinolone [85.7% (46.7–99.5)] and 3GC [93.1% (77.0– blunted by the L-type Ca2+ channel blocker nifedipine (0.8 Æ 2% KCl; P < 0.01). 99.1)] complied with national guidelines. However, 51.7% (34.4–68.6) of 3GC Microparticle-induced contraction was attenuated by the anticeramide antibody prescriptions and 57.1% (25.0–84.2) of quinolone prescriptions were avoidable. (2.4 Æ 2.1%) and by D-609 (1.5 Æ 1.7 %), but not by GW4869 (7.8 Æ 1.7%) Discussion: 3GC and fluoroquinolones are frequently prescribed for lower respi- as compared to parallel controls (12 Æ 2% KCl). Similar to microparticles, FasL- ratory tract infections in the ED. Half of these prescriptions may be avoided. Anti- induced contraction (14.5 Æ 3.9%) was attenuated by the anticeramide antibody biotic restriction policy should be implemented in our hospital in order to (1.6 Æ 1.6%) and by D-609 (3.8 Æ 2.2%), but not by GW4869 (7.2 Æ 1.9%). promote prudent use of 3GC. Microparticles also increased ROS in human PASMC in culture and the contrac- Keywords: Lower respiratory tract infections, Third generation cephalosporin, tile effect in PA was abolished by the ROS scavenger tiron (3 Æ 1% vs. 15 Æ 3% Fluoroquinolones, Antibiotic restriction. in parallel control). Microparticles expressed FasL and PA expressed Fas. Conclusion: Microparticles from apoptotic T lymphocytes produce pulmonary vasoconstriction involving the activation of Fas, acid sphingomyelinase, increase in ROS and inhibition of Kv channels. Keywords: Microvesicles, oxydative stress, vascular smooth muscle cells.

01-12 endoplasmic reticulum is involved in glycosylation, formation of disulfide bonds, Effects of budesonide on repeated cadmium inhalation-induced lung folding, and assembly of synthesized proteins. Indeed, proteins must be folded into inflammation and airspace enlargement in rats proper conformation by the endoplasmic reticulum in order to be fully efficient W Zhanga, J Zhia,ZXua, Y Cuia, Y Zhanga, J Habyarimanab, C Cambierb, and misfolded proteins are not delivered to Golgi apparatus. The accumulation of P Gustinb aFaculte de Medecine, Universite JiaoTong de Shanghai, Shanghai, China; unfolded or misfolded proteins induces endoplasmic reticulum dysfunction leading bFaculte de Medecine Veterinaie, Universite de Liege, Liege, Belgium to cell survival reduction. Numerous types of cellular stress such as hypoxia, As one of the components of particles involved in indoor pollution and particu- gene mutation and oxidative stress induce impairment of endoplasmic reticulum larly in cigarette smoke, cadmium has been shown to be related with obstructive function, leading to the so-called endoplasmic reticulum stress which subse- lung diseases and lung cancers. The aim of this study was to investigate whether quently activates a complex signaling network called the unfolded protein a pretreatment with budesonide exerts a protective effect against cadmium- response. The unfolded protein response leads to inhibition of protein synthesis induced lung inflammation and airspace enlargement. Rats were randomly and activation of protein degradation. The unfolded protein response has already assigned to a sham group and a cadmium group (n = 3/group) exposed to vehi- been associated with a large number of human diseases including cardiovascular cle (0.10% DMSO in saline) for 15 min immediately followed by an inhalation of disorders. Indeed, endoplasmic reticulum stress impact most cardiac functions, 0.1% CdCl2 solution for 60 min three times a week during 5 weeks respectively. including energy metabolism and cardiogenesis leading to cardiomyopathies, and In budesonide-pretreated groups (n = 3/group), rats were pretreated with budeso- heart failure. We have recently shown that pharmacological endoplasmic reticu- nide (250 lg/15 mL or 500 lg/15 mL) for 15 min followed by 60 min inhala- lum stress inhibition reduced hypertension associated cardiac hypertrophy and tion of saline or 0.1% CdCl2. The effects of these two different concentrations of fibrosis (reduction in collagen I content and transforming growth factor-b1 activ- budesonide on cell counts in bronchoalveolar lavage fluid (BALF) and on the ity). The vascular endothelium is also targeted by endoplasmic reticulum stress amount of cytokines (IL-1b, TNF-a and IL-8) in lung tissue homogenates with although less is known about the mechanism(s) linking the unfolded protein lung histomorphometry were investigated. Compared with the sham group, a sig- response to endothelial dysfunction. Nevertheless, a large body of evidences nificant increase in total cells count (0.85 Æ 0.25 vs. 0.23 Æ 0.13 9 106 cells/ shows a link between endoplasmic reticulum stress and endothelial dysfunction mL, P < 0.05) and neutrophils count (0.46 Æ 0.17 vs. in diabetes mellitus. Several recent studies suggest a possible link between hyper- 0.01 Æ 0.008 9 106 cells/mL, P < 0.05) was observed in cadmium group. Cad- glycemia-induced endoplasmic reticulum stress and oxidative stress inducing mium also induced a significant increase in the cytokine contents in lung tissue endothelial dysfunction. homogenates. The level of IL-8, TNF-a and IL-1b in cadmium group was higher than that of the sham group (16.44 Æ 1.88 vs. 10.23 Æ 0.67 ng/g, P < 0.01; 18.89 Æ 1.91 vs. 12.68 Æ 0.73 ng/g, P < 0.05; 17.56 Æ 0.99 vs. 02-01 10.01 Æ 1.09 ng/g, P < 0.01). The parenchyma inflammatory cell infiltration Mechanisms involving serotonergic 5-HT2B receptors in drug-induced and airspace enlargement interpreted by the mean linear intercept enhancement valvulopathy: pharmacological approach in mice (54.04 Æ 9.8 vs. 77.82 Æ 1.98 lm; P < 0.05) were detected in rats exposed to R Lawsona, B Gasserb, E Ayme-Dietricha, L Maroteauxc, L Monassiera aLaboratoire cadmium. The pretreatment of budesonide induced no effects on the increase of de Neurobiologie et Pharmacologie Cardiovasculaire, Strasbourg, France; bService de cell counts and cytokine levels, neither on the airspace enlargement induced by Pathologie, CHU Emile Muller, Mulhouse, France; cInstitut du Fer a Moulin, INSERM cadmium. In conclusion, while the high concentration of budesonide tested in U839, Paris, France this study (500 lg/15 mL) has been shown to exert an inhibitory effect on the Introduction: Recent safety data emphasized the close link between therapeutic neutrophilic infiltration in the cadmium-induced acute pulmonary inflammation, use of 5-HT2B serotonergic receptor agonists and cardiac valve degeneration (1) the preventive effect against repeated Cd exposures-induced inflammation and but the pharmacological mechanisms have not been clearly understood. Histo- remodelling is very limited. This model could be useful to investigate the mecha- pathological analysis of the lesions shows cell proliferation with extracellular nisms of resistance to corticoids. matrix deposition that leads to valvular leaflets stenosis driving to surgical valve Keywords: cadmium inhalation, chronic pulmonary inflammation, airspace replacement. In this work, we evaluated the effects of chronic administration of enlargement, budesonide. nordexfenfluramine (NDF) a highly selective 5-HT2B receptor agonist, on mice mitral valve leaflets. Methods: Adult male 129sv mice (aged 12 weeks) were randomly assigned in 01-13 six different groups and underwent a subcutaneous micro-osmotic pump implan- Characterization of the expression and the role of bitter taste receptors tation delivering either NDF (1 mg/kg/day) or vehicle (sterile water) for 4 weeks. in human lung parenchyma and macrophages Four groups were designed with wild-type mice: (i) sham-operated, (ii) NDF, (iii) S Grassin Delylea, S Taleb-Fayada, E Nalinea, C Abriala, M Brolloa, C Faisyb, NDF + L-NAME (300 mg/kg/day) in drinking water, (iv) NDF + ritanserine a a a b À/À C Magali , D Philippe Hôpital Foch, Suresnes, France; Hôpital Europeen Georges (2 mg/kg/day) in food and two groups of knockout mice (5-HT2B ): (v) sham- Pompidou, Paris, France operated and (vi) NDF. Mice were monitored by regular body weight, food and Introduction: Bitter-taste receptors (TAS2Rs) are a family of GPCRs including water intake, blood pressure measurement and echocardiography. 25 members in humans, whose expression in peripheral blood leucocytes (PBL), Results: At the end of the experiment, urinary 5-HIAA was also monitored to airway epithelial cells or smooth muscle cells has been recently documented. appreciate serotonin synthesis. Hearts were harvested for histological analysis to Their stimulation is responsible for relaxation in airways, and inhibition of cyto- quantify mitral valve leaflets thickness and cellularity. The comparison between kine production in PBL. The current study was performed to assess the expres- wild-type sham-operated and NDF treated mice didn’t reveal any significant dif- sion and the function of bitter taste receptors in the cytokine production of ferences in terms of food or water intake, body weight, echocardiography param- human lung parenchyma and macrophages. eters, beside of a slight blood pressure increase from 124 Æ 6 mmHg vs. Methods: Parenchyma explants (PE) and lung macrophages (LM) were isolated 141 Æ 10 mmHg (P < 0.05) for NDF treated mice at the mid term of the experi- from patients undergoing surgery for lung carcinoma, and then challenged with ment. We also point out a significant increase of urinary amount of 5-HIAA selected TAS2Rs agonists (1 lM–1mM), in the presence or absence of lipopolysac- (52 Æ 7 mg/L vs. 74 Æ 4 mg/L; P < 0.05). Histological analysis revealed charide (LPS). Agonists were selected on the basis of their known selectivity increased mitral valve leaflets thickness, and cell number (endothelial or intersti- towards the different receptor subtypes. Transcript expression of eight TAS2Rs tial) in NDF group (+20% over controls). These histological lesions were pre- was assessed with RT-qPCR and chemokine (TNF-a, CCL3 and CXCL8) produc- vented in part by a co-treatment with ritanserine and totally by L-NAME tion was measured with ELISA. showing the contribution of the eNOS. À/À Results: Transcript expression of TAS2R4, 5, 10, 14, 31, 39, 43 and 45 was Conclusion: Transgenic KO 5-HT2B mice did not show any response to NDF found in LM. Chloroquine (agonist for TAS2R3, 7, 10 and 39), quinine (4, 7, 10, stimulation indicating the crucial role of 5-HT2B receptors. Experiments are cur- 14, 39) and phenanthroline (5) induced a pronounced inhibition of the LPS- rently evaluating cellular and molecular mechanisms. induced TNF-a, CCL3 and CXCL8 production without affecting basal production References: and cell viability, whereas diphenidol (1, 7, 10, 14, 31, 39, 43…), denatonium (4, 1. Ayme-Dietrich E., Lawson R., Gasser B., Dallemand R., Bischoff N., Monassier 7, 8, 10, 13, 14, 39, 43), saccharin (8, 31, 43) and colchicine (4, 39, 46) were L. ‘Mitral Bioprosthesis Hypertrophic Scaring and Native Aortic Valve Fibrosis devoid of effect. In PE, quinine, chloroquine and phenanthroline abolished basal During Benfluorex Therapy’. and LPS-induced cytokine production. The LPS-induced production was partially Keywords: serotonin, valvulopathy, 5-HT2B receptors. altered by ofloxacine (9) and strychnine (7, 10, 46). Diphenidol, denatonium, saccharin and colchicine were also devoid of effect. In LM and PE, percentage inhibitions were between 50% and 100% for the highest concentrations applied. 02-02 Conclusion: The stimulation of TAS2Rs expressed in human PE and LM induced Endothelial protein tyrosine phosphatase 1B deficiency reduces both a dramatic inhibition of cytokine production. The lack of specificity of the avail- endothelial and cardiac dysfunction of in a mouse model of heart failure able agonists and the absence of selective TAS2R antagonists did not enable to J Maupointa, N Bouhzama, E Gomeza, M Besniera, JP Henrya, P Muldera, characterize the bitter taste receptors involved. However, these preliminary data V Richarda aInserm U1096, Rouen, France suggest that these receptors may constitute a new target for the pharmacological Chronic heart failure (CHF) is associated with endothelial dysfunction, involving management of inflammatory lung diseases. a decrease in nitric oxide (NO) production. However the direct link between endo- Keywords: Bitter taste receptors, cytokines, human lung macrophages, human thelial dysfunction and aggravation of CHF has not yet been established. Our lung parenchyma. group has recently revealed a new potent therapeutic approach of CHF, based on inhibition of protein tyrosine phosphatase 1B (PTP1B), which both increases NO production (via restored PI3K/Akt/eNOS signaling) and reduces adverse Left Ven- tricular (LV) remodeling and LV dysfunction. To address the direct link between SESSION 2: GS MIXTE CARDIOVASCULAIRE restored endothelial function and reduced CHF, we designed the present study to evaluate the cardiac and vascular consequences of endothelial PTP1B deficiency (endoPTP1BÀ/À) in a mouse model of CHF. Endoplasmic reticulum stress and vascular endothelium dysfunction We developed endoPTP1BÀ/À mice by crossing LOX-P PTP1B mice with mice D Henriona aLaboratoire de Biologie Neurovasculaire et Mitochondriale Integree, – expressing CRE recombinase controled by the endothelial promoter Tie2, or wild- INSERM U1083 CNRS UMR 6214, Angers, France type (WT) were subjected to left coronary artery ligation or sham surgery, and In eukaryotic cells the endoplasmic reticulum is formed in continuity with the the development of CHF was assessed by echocardiography at 2 weeks, and 1, 2 outer membrane of the nuclear envelope. The endoplasmic reticulum is the site and 3 months. of protein synthesis and maturation as well as lipid biosynthesis. The endoplas- WT CHF mice showed marked impaired flow-mediated dilatation of isolated mes- mic reticulum has also a major role in intracellular calcium storage. Thus, the enteric arteries (sham: 40 Æ 4%; CHF: 5 Æ 5; P < 0.001), which was improved

in endoPTP1BÀ/À mice (30 Æ 5%; P < 0.001 vs. WT CHF). These responses were 02-06 significantly reduced by a NO-synthase inhibitor, showing the implication of NO. Circulating microparticle levels and vascular function in a mouse model In WT, CHF increased LV diameters and decreased LV fractional shortening of combined intermittent hypoxia and high-fat diet (LVFS; at 3 months: sham: 49 Æ 2; CHF: 12 Æ 1%; P < 0.001) while these car- W Trzepizura,b, G Abderahima, C Arnaudc, C Ribuotc, P Levyc, diac parameters were improved in endoPTP1BÀ/À mice (LVFS: 20 Æ 2%; R Andriantsitohainaa, Mc Martineza, F Gagnadouxa,b aINSERM U1063, LUNAM P < 0.01). Left ventricular pressure-volume curves showed that endoPTP1BÀ/À Universite, Angers, France; bDepartement de Pneumologie CHU, Angers, France; mice with CHF had reduced LV end-systolic (sham: 22.5 Æ 1.2%; CHF WT: cINSERM UMR 1042, Universite de Grenoble-I, Grenoble, France 14.9 Æ 1.9%, P < 0.05) and increased end-diastolic pressure-volume relation- Introduction: There is a clinical co-incidence of the metabolic syndrome (MS) ships (sham: 2.1 Æ 0.9%; CHF WT: 4.9 Æ 0.7%, P < 0.05) demonstrating and the obstructive sleep apnea syndrome (OSAS), and each syndrome is associ- impaired systolic and diastolic dysfunction, respectively, which were both ated with endothelial dysfunction and cardio-vascular diseases. Microparticles improved in endoPTP1BÀ/À mice (end-systolic: 20.6 Æ 2.3%, P > 0.05; end-dia- (MPs) are submicron membrane vesicles, released into the extracellular space fol- stolic: 2.3 Æ 0.9%, P < 0.05). Histological analysis showed cardiomyocytes lowing to cell activation and implicated in the vascular damage associated with hypertrophy and increased collagen density in WT CHF mice, which were both syndromes. reduced in endoPTP1BÀ/À mice, at identical infarct size. The aim of this study was to evaluate the specific effects of intermittent hypoxia Thus, endothelial PTP1B deficiency not only induced an improvement of endo- (IH), obesity and the combination of both conditions on MPs levels and endothe- thelial function, but also improved cardiac function and reduced adverse LV lial function in mice. remodeling. These results provide a direct demonstration of the beneficial effect of Methods: Fifty two mice were divided in four groups: control group (normoxia endothelial protection in the treatment of heart failure. and normal diet), high-fat diet (HFD) (diet with 42% calories from fat for Keywords: Chronic heart failure, endothelial PTP1B, mice. 8 weeks) receiving group, IH group [30 s hypoxia (hypoxic plateau at 5% FiO2) followed by 30 s normoxia (normoxic plateau at 21% FiO2), 8 h a day, during 14 days] and a group receiving both HFD and IH. Body weight evolution was 02-03 evaluated for 8 weeks, and then, animals were sacrificed. Membrane surface and intracellular ultrastructural differences in Results: We showed that HFD was able to induce early-onset obesity as reflect- control and mdx cardiomyocytes highlighted by SICM and FFT ing by the increase of adipose tissue weight, dyslipidemia and a decrease in the C Lorina, E Aguettaza, A Krzesiaka, P Boisa, JF Faivrea, C Cognarda, endothelium-dependent relaxation evoked by acetylcholine. Exposure to IH alone S Sebillea aSTIM FRE 3511 Universite de Poitiers/CNRS, Poitiers, France was responsible for an increase in the rate of leukocyte and macrophage MPs, The aim of the study was to determine whether membrane surface and intracel- without changes in endothelium-dependent relaxation. Finally, animals submitted lular architecture of mdx (mouse model of Duchenne muscular disease – DMD) to the combination of both treatments displayed the same MPs profile than IH cardiomyocytes are different from those of normal mice. It is known that a treatment, associated with an insulin resistance as accessed by HOMA index. Sur- dilated cardiomyopathy is associated in humans with DMD and that a significant prisingly, intermittent hypoxia prevented endothelial dysfunction induced by HFD fraction of DMD patients die of cardiac function alteration. It was therefore alone. important to know whether there are differences in the organization of mem- Conclusion: This study suggests a specific role of intermittent hypoxia in the brane structures (membrane surface, transverse tubules, sarcoplasmic reticulum increased leukocyte-derived MP levels observed in OSAS. HFD alone induce an and associated proteins) at the origin of calcium movements and excitation-con- alteration of the vascular function but no increase in MPs. HFD and IH together traction coupling which are essential mechanisms in cardiomyocytes for the induce insulin resistance but a vascular protection as compared to HFD alone. heart physiological/pathophysiological contractile function. These results suggest that intermittent hypoxia might be considered as a precon- This study used an innovative technique (Scanning Ion Conductance Microscopy) ditionning mechanism inducing protection of the vascular function in HFD mod- to image, with a high resolution, the membrane surface (topography), as well as els. immunolabelling and FFT analysis to identify irregular positioning of involved Keywords: sleep apnea, metabolic syndrome, intermittent hypoxia, microparti- membrane and protein network structures. cles, vascular function. The SICM device has been calibrated and the image processing protocol was tuned up to be non-destructive. Then, in mdx, topographical images of cardiomyocytes were recorded. They shown disorganizations of the regular crest/groove 02-07 alternation pattern, observed in control cells, and the presence of relief accidents New genes and biological processes regulated by aldosterone- (‘pot-holes’). Immunolfluorescence images and FFT analysis and revealed that mineralocortico€ıd receptor complex in the heart. this is accompanied by the disruption of the regular disposition of the transverse B Graveza, S Messaoudia, C Latouchea, V Pellouxa, A Tarjusa, C Delcayreb, tubules and dihydropyridine receptors but not of the sarcoplasmic reticulum net- JL Samuelb, K Clementa, N Farmana, F Jaissera aINSERM U872, Paris, France; work and ryanodine receptors. bINSERM U942, Paris, France Conclusion: the absence of dystrophin in mdx could be at the origin of these Background: The mineralocortico€ıd receptor (MR) and its ligand, aldosterone alterations. The work will be pushed forward to study these properties under con- (aldo), are involved in cardiac diseases, like myocardial infarction and atrial fibril- ditions where cardiomyocytes will be mechanically stretched using carbon fibers. lation. Deleterious consequences of enhanced mineralocorticoid signaling has Keywords: DMD, mdx, membrane topography, SICM, FFT, cardiomyocytes, dys- been highlighted by clinical trials (RALES, EPHESUS) showing a major benefit of trophin, excitation-contraction coupling. MR antagonists on top of standard treatment in patients with heart failure. Car- diomyocyte-MR regulated processes leading to these injuries are still unclear. Objective: To identify new aldosterone-MR regulated functions and genes possi- 02-04 bly involved in cardiac diseases. Cardio-pulmonary exercise echocardiography test (cpeet): a new exercise Methods: Transgenic mice overexpressing MR in cardiomyocytes (MR-Cardio) test to explore cardio-pulmonary pathophysiology and their control littermates (Ctrl) were treated for 7 days either with aldo D Guijarroa, N Pirioua, B Mottina, P Jaafara, JP Gueffeta, D Kerdoncufa, (60 lg/kg/day; n = 6–9/group) or vehicle (physiological serum), and a genome JN Trochua, T Le Tourneaua aInstitut du Thorax, Inserm U1087 – CHU de Nantes, microarray analysis was performed. Nantes, France Results: Microarray analysis on heart of MR-Cardio and Ctrl mice. Eight hundred Background: Exercise echocardiography (Ex-Echo) and cardiopulmonary exer- and sixty-five genes were found differentially regulated. Among them, 165 genes cise test (CPET) are essential to investigate complex cardiac or pulmonary pathol- were up-regulated by both aldo and MR overexpression. Connective tissue growth ogies. Both tests, performed separately, are helpful for the diagnostic, prognostic factor (CTGF) was one of the most aldo-MR regulated genes. Validation of CTGF: and therapeutic assessment in numerous clinical situations. The qPCR analysis in the heart of mice showed that the CTGF expression was Objectives: To evaluate a new exercise test combining Ex-Echo and CPET. Car- increased in MR-Cardio-vehicle mice compared to Ctrl-vehicle mice (three times; diopulmonary exercise echocardiography test (CPEET) aims to improve our P < 0.05). This expression was further increased by aldosterone treatment (2.5 approach and understanding of complex cardiac and/or pulmonary diseases. times; MR-Cardio+aldo vs. MR-cardio-vehicle; P < 0.05). The immuno-staining of Methods: Patients referred to our department for evaluation of complex valvular heart sections showed that CTGF was produced specifically by cardiomyocytes. In diseases, severe cardiomyopathy or exertional dyspnea of unknown origin carried H9C2-MR+ cell line, aldosterone induced a dose-dependent increase of CTGF out a CPEET during their clinical work-up. A comprehensive Ex-Echo was com- expression (2.5 times; P < 0.05) that was prevented by MR specific antagonist bined to a cardiopulmonary evaluation (according to their current guidelines) for spironolactone. A Gene ontology analysis was realized on the genes from micro- one single cyclo-ergometric exercise in semi-supine position. Exercise parameters array, with Genespring software, in order to identify the biological processes mod- including workload, heart rate and blood pressure changes, ECG, rest and exer- ulated by aldo-MR complex. The 70 % of identified genes were involved in cell cise echocardiography parameters, and gas exchanges were analyzed. cycle. Cellular proliferation: The immuno-staining of heart showed an increase of Results: Twenty-five patients (57 Æ 2 years, 13 men) carried out the CPEET. colocalisation in endothelial cells of Ki-67 (a proliferation marker) and caveolin-1 Indications of CPEET were mitral valve diseases (12 patients, 48%), aortic valve (an endothelial cells marker) in MR-Cardio+aldo mice. disease (three patients, 12%), cardiomyopathy (six patients, 24%) and exertional Conclusion and perspectives: Our results show two different and specific roles dyspnea of unknown origin (four patients, 16%). According to our preliminary of the aldo-MR complex in the heart: (i) the regulation of CTGF expression in car- experience, exercise echocardiography in the semi-supine position did not alter diomyocytes by aldosterone via MR activation; (ii) the increase of cardiac endo- respiratory devices function and gas exchange analyses. Exercise was ended for thelial cells proliferation. We are now studying, with HUVEC cells, the dyspnea or muscle exhaustion in 52% and 48% of patients, respectively. Maximal mechanisms underlying the proliferation of endothelial cells. workload was 115.0 Æ 9.8 watts; peak oxygen consumption averaged Keywords: aldosterone, mineralocortico€ıd receptor, heart. 19.7 Æ 1.3 mL/kg/min (77.8 Æ 3.9% of maximal predicted value). Combination of Ex-Echo and CPET was helpful in the diagnostic, prognostic or therapeutic evaluation in 18 patients (78%) and provided new insights in the understanding 02-08 of complex physiopathology. Associations between visceral adipose tissue, liver steatosis, Conclusion: CPEET is a promising test to perform a comprehensive evaluation of atherosclerosis and plasma osteoprotegerin levels in dysmetabolic adults complex cardiac situations in a single examination. Results of Ex-Echo were com- PH Ducluzeaua, J Boursierb, C Aubec, G Leftheriotisd aDiabetologie, CHU, Angers, forted by gas exchange analyses. Further studies are needed to validate cardiopul- France; bHepatologie, CHU, Angers, France; cRadiologie, CHU, Angers, France; monary data obtained in the semi-supine position compared to upright CPET. dExplorations Vasculaires, CHU, Angers, France Keywords: exercise echocardiography – cardiopulmonary exercise test. Context and objective: Obesity and the metabolic syndrome are accompanied by increased visceral adipose tissue as well as liver steatosis, both leading to increased cardiovascular risk. High plasma osteoprotegerin has been more

recently associated with cardiovascular diseases. However, less is known about brane protein disulfide isomerase (PDI), an enzyme involved in •NO release from the associations between visceral adipose tissue (VAT), liver fat content (LF) and S-nitrosothiols, were also evaluated using oxadiazolo [4,3-a] quinoxalin-1-one plasma osteoprotegerin (OPG) in dysmetabolic subjects. The objective is to deter- (ODQ), a sGC inhibitor, and bacitracin, a PDI inhibitor. mine the association between VAT, LF, and other markers of metabolic syndrome Results: Expressed as means Æ SEM, one way ANOVA test. Synthesized BUC(NO)2 (i.e. hypertension, dyslipidemia and insulin resistance) and OPG in dysmetabolic was structurally characterized, exhibited a high purity (99.5 Æ 0.1 %, n = 6 adults. batches) and was stable during 4 days when prepared in ultra-pure water and Methods and patients: Three hundred and nine subjects from the NUMEVOX stored at 4 °C, pH < 0.5. BUC(NO)2 induced a higher concentration-dependent cohort were included on the basis of at least one metabolic syndrome criterion. vasodilatation than the mixture of SNAP + NACNO (pD 7.8 Æ 0.1 and 2 • They underwent a thorough metabolic and cardiovascular evaluation including: 6.7 Æ 0.2, respectively P < 0.05). BUC(NO)2 induced a sixfold increase of NO arterial stiffness and atherosclerotic plaques, HOMA insulin resistance index content in the aorta as compared to basal value (107 Æ 1.6 mM), which was sig- (HOMA-IR), liver enzymes, ferritin, waist circumference and BMI. Plasma OPG nificantly higher than the •NO release from the mixture of SNAP + NACNO (four- was assessed by Elisa technique. VAT and LF were measured by magnetic reso- fold increase vs. basal value). The concentration response curves to BUC(NO)2 nance imaging in a subgroup of 140 patients. Ultrasound examination of arteries were significantly shifted to the right in the presence of ODQ 5 lM (pD2 and arterial stiffness (pulse wave velocity) were recorded. Age and gender 4.7 Æ 0.1) and bacitracin 100 lM (pD2 6.8 Æ 0.2). adjusted paired correlation were calculated. Conclusion: We successfully synthesized the new dinitrosothiol BUC(NO)2 with Results: BMI, waist circumference and MRI-based VAT correlated with OPG a satisfactory purity degree and stability. Our data showed that BUC(NO)2 (r = 0.19, P = 0.006; r = 0.18, P = 0.001; r = 0.24, P = 0.004 respectively) releases a large amount of •NO into the aorta, partly through PDI activation, while subcutaneous fat didnot correlated. Liver fat content correlated with OPG activates sGC and induces a potent vasorelaxation. (r = 0.25, P = 0.003) as well as ALT (r = 0.39, P < 0.001), gGT (r = 0.35, Keywords: S,S’-dinitrosobucillamine, physico-chemical characterization, stability, P < 0.001), ferritinemia (r = 0.18, P = 0.002) and triglyceridemia (r = 0.17, vasorelaxation, protein disulfide isomerase, rat aortic ring. P = 0.004). HOMA-R was strongly correlated with OPG (r = 0.39, P < 0.0001), as well as leptinemia (r = 0.28, P < 0.0001) and adiponectinemia (r = À0.12 P = 0.05).Plasma OPG correlated to arterial stiffness (r = 0.19, P = 0.04). In the 02-11 whole cohort OPG correlated weakly with the number of atherosclerotic sites Sunitinib hypertension: key role of the sympathetic nervous system (r = 0.12, P = 0.03). F Despasa, A Nesserb, JM Senardb, A Pathakb aCHU Toulouse, Toulouse, France; Conclusions: Plasma OPG levels were positively associated with VAT and liver bINSERM 1048, Toulouse, France steatosis, as well as insulin resistance and adipokines. The significant association Hypertension (HTN) is a major risk factor increasing cardiovascular morbi-mor- between OPG and some of the major components of the metabolic syndrome sug- tality. 10 % of HTN cases are drug-related. Anti-angiogenic therapy (AAT), used gests a possible link with the vascular complications associated to the metabolic in cancer treatment to limit tumor angiogenesis, has been reported to induce syndrome. HTN. The Bevacizumab, monoclonal antibody binding to circulating VEGF, and Keywords: osteoprotegerin, metabolic syndrome, atheromatosis. the tyrosine kinase receptor inhibitors (Sorafenib and Sunitinib) inhibit angiogenesis. HTN is a common complication of these drugs, which concern up to 30 % of patients within 3–10 weeks after the first exposure. But AA-induced HTN 02-09 mechanisms remain unclear. Several arguments allow suggesting the involve- Estimation of the pulsatile fraction of right ventricular stroke work in ment of the sympathetic nervous system (SNS) in AAT-induced HTN. First, trans- pulmonary hypertension genic mice expressing low levels of VEGF determines neurovascular synapsis D Chemlaa, V Castelainb, F Zhuc, Y Papelierc, N Creuzec, S Hoetted, F Parente, structural abnormalities associated with HTN. Moreover, essential HTN is often G Simonneaud, M Humbertf, P Herveg aUniversite Paris Sud-CHU de Bicêtre-APHP, associated with an increase of SNS activity. Finally, it has been shown that AA Le Kremlin-Bicêtre, France; bHôpitaux Universitaires de Strasbourg, Strasbourg, drugs increase arterial stiffness, which leads to sympathetic activation. France; cUinversite Paris Sud, Clamart, France; dUniversite Paris Sud, Le Kremlin- According to these data, our work was primarily aimed to study the SNS role in Bicêtre, France; eCHU de Bicetre-APHP, Le Kremlin-Bicêtre, France; fINSERM UMR AAT-induced HTN. We also characterized baroreflex gain in our experimental 999, Le Kremlin-Bicêtre, France; gCentre Chirurgical Marie-Lannelongue, Le Plessis- model. For this purpose, we performed sunitinib gavage (40 mg/kg/day) in a Robinson, France group of mice during 8 days compared with a placebo group. Blood pressure (BP) Background: The mean pulmonary artery pressure (mPAP) replaces mean sys- was measured by carotid artery catheterism and SNS activity was quantified by tolic ejection pressure (msePAP) in the classic formula of right ventricular stroke microneurography direct recording of sympathetic renal nerve activity (RSNA, work RVSW = (mPAP-RAP) 9 stroke volume, where RAP is right atrial pressure. bursts/s). Arterial baroreflex activity was assessed by vasoactive drugs injection: Only the steady work is thus taken into account, not the pulsatile work, whereas sodium nitroprusside, vasodilator (1 mg/kg) and Phenylephrine, vasoconstrictor pulmonary circulation is highly pulsatile. Our retrospective, high-fidelity pressure (20 mg/kg) to establish a slope determining the adaptation of the RSNA accord- study tested the hypothesis that msePAP was proportional to mPAP, and looked ing to BP variations. Finally, to confirm SNS role on BP, we evaluated adrenergic at the implications for RVSW. antagonist’s effects on hemodynamic and autonomic parameters during sunitinib Methods: Eleven patients with severe precapillary pulmonary hypertension (PH, treatment. six idiopathic pulmonary arterial hypertension and five chronic thrombo-embolic Systolic/diastolic BP significantly increased in sunitinib-treated mice (123 Æ 2/ PH, mPA P = 59 Æ 13 mmHg) were studied at rest and on mild-to-moderate 87 Æ 3 vs. 112 Æ 1/79 Æ 1 mmHg in control mice). Heart rate significantly exercise (cycling while supine). Eight non-PH controls were also studied at rest decreased in sunitinib-treated mice (553 Æ 16 vs. 606 Æ 5 beats/min in control (mPA P = 16 Æ 2 mmHg). The msePAP was averaged from end-diastole to mice). RSNA was significantly increased in sunitinib-treated mice (40 Æ 7 vs. dicrotic notch. 20 Æ 2 bursts/s in control mice). Baroreflex gain was significantly decreased in Results: In the full data set (53 pressure-flow points), mPAP ranged 14– sunitinib-treated mice (À1.1 Æ 0.2 vs. À1.8 Æ 0.2 %RSNA/mmHg). The effect of 99.5 mmHg, cardiac output 2.38–11.1 L/min and heart rate 53–163 bpm. sunitinib on BP and RSNA was totally prevented by prazosin (4 mg/kg/day). There was a linear relationship between msePAP and mPAP (r²=0.99). The mse- In conclusion, Sunitinib-induced HTN is related to an increase in SNS activity PAP matched 1.25mPAP (bias = À0.5 Æ 2.6 mmHg). Results were similar in and mediated by vascular alpha1-adrenoreceptor. The mechanism accounting for resting non-PH and in resting and exercising PH. This implies that the classic for- these findings, potentially involving increased arterial stiffness and/or chemore- mula markedly underestimates RVSW and that the pulsatile RVSW may be a flex activation are currently under investigation in our laboratory. variable 20–55% fraction of RVSW depending on RAP and mPAP. At rest, PH Keywords: Hypertension, antiangiogenics drugs, sympathetic nervous system. patients had two-times higher RVSW than non-PH (P < 0.001), but similar pulsatile work fraction (26 Æ 4% vs. 24 Æ 1%) because of the counterbalancing effects of high RAP (11 Æ 4 vs. 4 Æ 2 mmHg), which increases the fraction, and 02-12 high mPAP, which decreases the fraction. Increase in angiogenesis after myocardial infarction in protein-tyrosine- Conclusions: Our study favored the use of an improved formula which takes phophatase-1b (ptp1b) deficient mice into account the variable pulsatile work: RVSW = (1.25 mPAP À RAP) 9 stroke M Besniera, A Galaupb, L Nicola, JP Henrya, M Debunnea, A Guereta, volume. Increased RAP and increased mPAP have opposite effects on the pulsa- I Boukhalfaa, F Lallemanda, P Muldera, S Germainb, V Richarda, A Ouvrard- tile work fraction. Pascauda aU1096 Inserm, Rouen, France; bU1050 Inserm, Paris, France Keywords: Right ventricle. Systolic function. Pulmonary hypertension. Enhanced angiogenesis is a promising therapy for preserving heart function after myocardial infarction (MI). Angiogenesis is controlled by growth factors such as vascular endothelial growth factor (VEGF) whose receptors are activated by tyro- 02-10 sine autophosphorylation. The protein tyrosine phosphatase 1B (PTP1B) is a neg- S,S’- dinitrosobucillamine: a new nitric oxide donor acting as an efficient ative regulator of growth factors signaling. We therefore tested whether PTP1B vasorelaxant agent deletion in mice enhances angiogenesis after MI provoked by coronary ligation. F Dahboula, C Perrin-Sarradoa, A Boudiera, I Lartauda, R Schneidera, We compared infarcted hearts of PTP1BÀ/À and WT mice at 3, 8 days and P Leroya aUniversite de Lorraine, Nancy, France 3 months post-MI. At 3 days post-MI, capillary density was increased in Introduction: S-nitrosothiols are an emerging class of •NO donor drugs with PTP1BÀ/À mice in the border zone of the ischemic area (+9% vs. WT, P = 0.06). potential use in cardiovascular disorders associated with decrease in nitric oxide At 8 days and 3 months post-MI, capillary density was strongly enhanced in all (•NO) bioavailability. the left ventricle of PTP1BÀ/À mice (8 days: +18% & 3 months: +23% vs. WT, The aim of this study was to synthesize and characterize a new dinitrosothiol, ‘S, P < 0.01). Using Magnetic Resonance Imaging, we measured myocardial perfu- S’-dinitrosobucillamine’ (BUC(NO)2), which combines in its structure two mono- sion and coronary reserve, referred as the difference between the basal and the nitrosothiols: S-nitroso-N-penicillamine (SNAP) and S-nitroso-N-acetylcysteine maximal perfusion, obtained by injection of an agonist of A2a adenosine recep- (NACNO), and to compare their vasorelaxant properties. tor. Prior to MI, PTP1BÀ/À mice had a significantly greater coronary reserve than Materials and methods: After synthesis, BUC(NO)2 characterization and purity WT without any change in basal perfusion. However, 8 days after MI, while the were evaluated using UV-visible spectroscopy, mass spectrometry and Saville-Gri- coronary reserve was abolished, PTP1BÀ/À mice displayed a higher basal perfu- ess methods, and product stability was tested. The vasorelaxant effects were sion (PTP1BÀ/À vs. WT: 13.7 Æ 0.3 vs. 11.0 Æ 0.3 mL/mg/min, P < 0.001). À6 assayed in phenylephrine (10 M)-precontracted aortic rings (isometric method) We then studied some of the cellular and molecular events known to be associ- isolated from male adult Wistar rats (n = 6–12 rings per group). The release of ated with angiogenesis. At 3 days post-MI, we measured a higher proportion of • À/À NO from BUC(NO)2 into the aorta was measured using 4,5-diaminofluorescein M2 pro-angiogenic macrophages in the heart of PTP1B mice. Moreover, after diacetate. Effects of BUC(NO)2 were compared to an equimolar mixture of NACNO immuno-precipitation of the VEGF Receptor 2, we detected an increased phos- plus SNAP. Involvement of cytosolic soluble guanylate cyclase (sGC) and mem- phorylation level of the associated protein Src in hearts from PTP1BÀ/À mice,

together with an elevated dissociation of the VEGFR2/VE-Cadherin complex, Genetics and mechanisms of Gordon’s Syndrome which may participate to vessel plasticity. X Jeunemaitrea aPARCC – UMRS 970, Centre de Recherche Hôpital Europeen Finally, at 3 months post-MI, the increased capillary density was associated with Georges Pompidou – Universite Paris Descartes, Paris, France reduced myocardial hypertrophy and reduced fibrosis, as well as an increased LV PseudoHypoAldosteronism type 2 (PHA2) or Familial Hyperkalemic Hypertension function. (FHHt) is a fascinating Mendelian disorder that has allowed the identification of Our results indicate that PTP1B deletion in mice stimulates post-MI angiogenesis unsuspected proteins in the regulation of ion transport in the distal nephron. In in the viable myocardium. Events downstream of VEGFR2 participate to this 2001, specific gain-of-function mutations were identified in the WNK1 and increase in angiogenesis which is likely to contribute to a higher myocardial per- WNK4 genes. These genes were then demonstrated to play a crucial role in the fusion that benefits cardiac function. Na and K balance in the distal nephron, mainly through activation of the Na-Cl Keywords: PTP1B, angiogenesis, heart failure. cotransporter NCC. I will show the interest of transgenic mouse models to get further insight into the molecular mechanisms involved in WNK1 intron 1 deletion. The suppression of insulator and repressor sequences lead to reciprocal 02-13 influences of the proximal and renal promoters on the expression of the L-WNK1 Protein tyrosine phosphatase-1b deletion in mice improves cardiac and and KS-WNK1 isoforms. In 2012, two recent studies identified two other unex- endothelial functions during aging pected genes (KLHL3 and CUL3) as responsible for the disease. Mutations are M Besniera, D Coquerela, J Favrea, F Bauera, F Tamiona, V Richarda, A Ouvrard- found in patients with different modes of inheritance (dominant, recessive, de Pascauda aU1096 Inserm, Rouen, France novo) with genotype-phenotype differences. Kelch-like 3 and Cullin 3 proteins are Cardiovascular function declines with advanced age. Cardiovascular aging is part of an E3 ubiquitinase complex that play a crucial role in the degradation of associated with left ventricular (LV) hypertrophy, myocardial fibrosis and endo- multiple proteins by the proteasome after ubiquitylation. Thus, up to now, four thelial dysfunction, all contributing to a progressive decrease in heart function. genes are responsible for FHHt, that were previously unsuspected to play a role Our laboratory recently showed that Protein Tyrosine Phosphatase 1B (PTP1B) in renal ion homeostasis. Despite genetic and phenotypic heterogeneity, similar deletion in mice, confers myocardial and endothelial protection in the experimen- mechanisms may explain this rare Mendelian form of renal hypertension, espe- tal model of heart failure after myocardial infarction. However, whether PTP1B cially an overactivity of the Na-Cl cotransporter. deletion could slow aging-associated cardiovascular damages remains unknown. We assessed LV function by echocardiography in PTP1B deficient (PTP1BÀ/À) and wild-type (WT) mice aged 3, 12, 15 and 18 months. At the age of 03-01 18 months, LV function was assessed by invasive pressure-volume curves that WNK4 regulates electroneutral NaCl transport in the intercalated cells allow measuring LV end-systolic and LV end-diastolic pressure-volume relation- of the collecting duct ships (LVESPVR and LVEDPVR). We also evaluated mesenteric flow-mediated M Jayata,CBusst€ a, P Houillierb, D Eladaria, S Alperc, J Hadchouela, vasodilatation by arteriography and coronary function on a Mulvany-wire-myog- R Chambreyb aINSERM U970 Equipe 3, Paris Cedex 15, France; bINSERM U872 raph. Finally, histological analyses of heart remodeling were performed. Equipe 3, Paris, France; cRenal Division and Molecular and Vascular Medicine Unit, Echocardiographic analyses showed that the aging-dependent decrease in LV Beth Israel Deaconess Medical Center, Boston, MA, USA fractional shortening was less marked in PTP1BÀ/À mice, compared to WT Objectives: Gordon’s syndrome (also known as Familial Hyperkalemic Hyperten- (decrease at 18 vs. 3 months: WT: À54%; PTP1BÀ/À: À29%, P < 0.001). Fur- sion (FHHt), or Pseudohypoaldosteronism type II (PHAII)) is a rare Mendelian thermore, the aging-induced LV diastolic dysfunction evidenced by the modifica- form of human hypertension, associated with hyperkalemia and hyperchloremic tion of the echocardiographic trans-mitral Doppler E/A parameter was metabolic acidosis. In a subset of patients, the syndrome is due to mutations in significantly improved in PTP1BÀ/À mice. At the age of 18 months, which the WNK1 and WNK4 genes encoding two serine-threonine kinases of the WNK approaches the limit of life expectancy of these mice from Balb/c genetic back- (With No lysine (K)) family. The patients are particularly sensitive to thiazide ground, PTP1BÀ/À mice showed a higher LVESPVR (WT vs. PTP1BÀ/À: diuretics, blockers of the Na-Cl cotransporter NCC. Accordingly, the analysis of a 13.9 Æ 0.9 vs. 18.4 Æ 1.6 mmHg/RVU, P < 0.05) and a lower LVEDPVR (WT transgenic PHAII model (TgWNK4PHAII), overexpressing a mutated form of vs. PTP1BÀ/À: 5.1 Æ 0.8 vs. 1.2 Æ 0.3 mmHg/RVU, P < 0.01), respectively WNK4, revealed an increase in NCC expression and activity. However, others highlighting improvement in LV systolic and diastolic functions. Moreover, con- studies have shown that this upregulation is not sufficient to explain the pheno- sidering the function of arteries in aged mice, the deletion of PTP1B partially type. We have previously demonstrated the presence of amiloride-resistant, thia- restored flow-mediated dilatation of isolated mesenteric arteries (% dilatation to zide-sensitive electroneutral NaCl absorption in the cortical collecting duct, flow at 200 lL/min; WT vs. PTP1BÀ/À: À0.4 Æ 2.1 vs. 7.4 Æ 2.6, P < 0.05) resulting from the coupling of the Na+-driven ClÀ/HCO À exchanger (Ndcbe/ À À 3 and slightly improved coronary artery relaxations in response to insulin (% relax- Slc4a8) with the Cl /HCO3 exchanger (pendrin/slc26a4) in the apical mem- À6 À/À ation at 10 M: WT vs. PTP1B 15 Æ 4 vs. 24 Æ 3). Finally, cardiac hyper- brane of intercalated cells. Our purpose was to characterize the regulation of trophy and fibrosis that develop with aging were significantly reduced in NaCl absorption by WNK4 in intercalated cells and its contribution to the PHAII PTP1BÀ/À mice that also displayed a higher myocardial capillary density. phenotype. Taken together, these results show that PTP1B deletion in mice improved cardiac Material and results: We first measured the transepithelial fluxes in isolated and endothelial functions in aging. microperfused CCD and showed that NaCl absorption was strongly stimulated in Keywords: PTP1B, aging, cardiovascular. TgWNK4PHAII mice and fully inhibited by luminal addition of thiazide (n = 5 tubules in each group). Accordingly, Ndcbe protein expression was upregulated in TgWNK4PHAII mice. Quantification of pendrin activity in microperfused CCDs evidenced an increase in TgWNK4PHAII micecompared to controls À À SESSION 3: GS REIN ET MILIEU INTERIEUR (2.9 9 10 3Æ0.37 pH units/s, n = 4 tubules vs. 5.2x10 3Æ0.64 pH units/s, n = 7 tubules, P < 0.05). We also observed an increase in the number of pendrin-positive intercalated cells in the CCD of transgenics mice (37.13 Æ 3.64%, Hereditary diseases of collagen IV: from mice to humans = Æ = a a n 1867 ICs from four control animals vs. 49.68 2.01%, n 1955 ICs from E Plaisier Service de Nephrologie et Dialyses, INSERM UMRS 702, Hôpital Tenon, four mutant mice, P < 0.05). Finally, in vitro studies showed that the WNK4- Universite Pierre et Marie Curie, Paris 6, France PHAII mutant stimulates pendrin activity in Xenopus laevis oocytes Basement membranes (BM) are highly specialized extracellular matrix that is (10.47 Æ 1.94 vs. 28.13 Æ 0.97 mmol/oocyte/h, n = 10 in each group, found in nearly all organs, including kidney, muscle, vessels and central nervous P < 0.001). system. They are playing a structural role, but are also involved in organogene- Discussion: These results suggest that WNK4 is an important regulator of NaCl sis, cell migration differentiation and proliferation. Collagen IV is one of the main reabsorption by the CCDs, through the modulation of the electroneutral thiazide- BM components. The a1(IV) and a2(IV) chains, encoded by COL4A1 and COL4A2 sensitive Ndcbe/Pendrin pathway in the intercalated cells. genes respectively, are the most ubiquitously expressed collagen IV isoforms. Het- Keywords: WNK4, Ndcbe/pendrin, NaCl transport, cortical collecting duct, inter- erozygous mutations affecting both genes are responsible for a small vessel brain calated cells. disease in human, sometimes associated with various eye defects. Within COL4A1-related diseases, a distinct phenotype, named HANAC (hereditary angiopathy, nephropathy, aneurysms and cramps) is also observed. In HANAC, a sys- 03-02 temic vasculopathy, that also included a brain angiopathy, is associated with Calcium-induced urinary acidification involves GPRC6A renal defects (larges cysts, chronic renal failure, sometimes hematuria), and a ML Figueresa, A Loupya, S Baronb, SK Ramakrishnana, B Wootlaa, C Mandetc, muscle disease with cramps and CPK elevation. Vascular investigations in HA- P Brunevalc, S Smajilovicd,HBrauner-Osborne€ d, P Houilliera aCentre de Recherche NAC patients showed that collagen IV affects not only structural but also func- des Cordeliers, Unite INSERM UMRS872, Paris, France; bService d’explorations tional properties of vessels. Contrasting with the patients presenting a brain- fonctionnelles renales, Hôpital HEGP, Paris, France; cService d’anatomopathologie restricted vascular disease, COL4A1 HANAC mutations affect a small region of renale, Hôpital HEGP, Paris, France; dDepartment of Drug Design and Pharmacology, the protein that contain major binding site for integrins, suggesting a potential Faculty of Health and Medical Sciences, Copenhague, Denmark phenotype-genotype correlation. Several Col4a1 mutant mice are available and Introduction: Urinary pH decreases when urinary calcium (Ca) concentration allow the analysis of pathomechanisms of organs involvement. Col4a1 HANAC increases; this is thought to protect against the formation of Ca-containing crys- mice study highlights a previously undiscovered role of Col4a1 during renal tals in collecting ducts (CD). However, the mechanism of Ca-sensing in the corti- embryogenesis and confirms the role of BM in the vasodilation and vasoconstri- cal CD (CCD) remains elusive. We hypothesized that GPRC6A, a G protein- tive properties of vessels, previously observed in patients. In addition, analysis of coupled receptor, may be involved in this phenomenon. The aim of our project the muscle phenotype suggests a primary role of microvascular defects in the was first to localize GPRC6A expression in the mouse kidney, then to assess its occurrence of the muscle fiber alteration and symptoms. This animal model will role on the Ca-dependent urinary acidification in vitro and in vivo. be very useful to test several pharmacological approaches. Methods: GPRC6A localization in mouse kidney was assessed by immunohisto- Keywords: Rein. chemistry. Antibodies risen against NCC, THP, AQP2 and CaSR were used to determine its precise localization. In in vitro microperfused CCDs, H+ secretion by type A intercalated cells was assessed by monitoring the rate of intracellular pH recovery (using the fluorescent pH-sensitive dye BCECFâ), after acidifying the + cells by a NH3/NH4 prepulse method. In vivo, urinary pH and Ca under basal conditions, DiHydroTachysterol (DHT)-induced hypercalciuria or acid load À/À +/+ (NH4Cl), were measured in Gprc6a and Gprc6a mice.

Results: The apical membrane of CD cells was stained by the anti-GPRC6A anti- 03-05 body, both in cortical and medullary CD, whereas no staining was seen with an Notch3 receptor involvement in experimental crescentic anti-CaSR antibody. The luminal presence of a GPRC6A ligand (3 mM Ca or glomerulonephritis + a a a a a 1mM ornithine) increased the rate of H secretion by type A intercalated cells in F El Machhour , L Mesnard , C Chatziantoniou , JC Dussaule INSERM U702, CCDs from Gprc6a+/+, but not from Gprc6aÀ/À mice. In vivo, urinary pH and Ca Paris, France were similar in both groups of mice under basal conditions. DHT-induced hyper- Background: Chronic kidney disease (CKD) is highly prevalent in the general calciuria elicited a significant urinary acidification in Gprc6a+/+ but not in population and glomerulopathies represent the third cause of end stage renal dis- Gprc6aÀ/À mice. No difference in the ability to decrease the urinary pH during an ease. Notch3 belongs to the well-known family of Notch receptors. It is involved oral acid load was noted between Gprc6aÀ/À and Gprc6a+/+ mice. in the development of resistive arteries and of glomeruli. Very little is known Conclusion: In the mouse CD, GPRC6A, but not CaSR, is expressed at the apical about the de novo activation of Notch3 in adult tissues under pathological condi- membrane. Increasing luminal concentration of GPRC6A agonists increases H+ tions. Our work aimed to study the involvement of Notch3 receptor in the mech- secretion by type A intercalated cells, an effect that involves GPRC6A. By opposi- anisms of progression of experimental crescentic glomerulonephritis (GN). tion, metabolic acidosis also activates H+ secretion, but does not involve Methods: Notch3 knock out (KO) Sv129 mice and their wild-type (WT) litter- GPRC6A. mates were treated by nephrotoxic serum (NTS) to induce GN. Two control Reference: groups received injections of the vehicle. Mice were sacrificed after 9 days and Wellendorph P., et al. No evidence for a bone phenotype in GPRC6A knockout tissue, urine and plasma samples were collected and used for subsequent mice under normal physiological conditions. J. Mol. Endocrinol. (2009). analysis. Keywords: Hypercalciuria, Urinary acidification, GPRC6A, Intercalated cells. Results: After 9 days of GN progression, the expression of Notch3 transcript was significantly upregulated in the whole cortex of WT mice and the expression of this receptor was induced in glomeruli, especially in podocytes. The two groups 03-03 developed GN but Notch3 KO mice were relatively protected compared to WT as Plasma cystatin C is a valid marker of glomerular filtration rate in mice demonstrated by the decreased values of plasma urea (21.3 Æ 2.2 vs. N Mayeura, A Jaafara, M Buleona, F Praddaudea, M Valleta, I Tacka aLaboratoire 27 Æ 1.3 mmoL, P < 0.05) and of proteinuria (11.6 Æ 2.2 vs. 16.1 Æ 1.3 g/ de Physiologie et INSERM UMR1048, Toulouse, France mmol P < 0.05). The improvement of renal function was associated with fewer Determination of the glomerular filtration rate (GFR) is ideally performed by mea- deposits of fibrin within glomeruli, with a reduced extracapillary cell proliferation suring the renal clearance of exogenous markers such as inulin and EDTACr51. and with the preservation of nephrin expression. The conclusion about the In mice, this technic present shortcomings: a high level of pratical expertise, a involvement of Notch3 receptor in the inflammatory response was reinforced by prolonged time of procedure and animal anesthesia. Thus, for years, the follow the blunted expression of MCP-1 and VCAM-1mRNA, the decrease of expression up of renal function has been based on the dosage of blood markers, such as of p65 NFkB subunit protein and the reduction of interstitial macrophage infiltra- creatinin and urea. However, various works have underlined the limitations of tion in the kidneys of Notch3 KO mice. Our in vitro studies showed that the acti- these markers. As a consequence and despite recent technical advances a reliable vation of Notch3 receptors in podocytes was associated with a migratory and easy to use endogenous marker to estimate GFR is still lacking in mice. phenotype. Cystatin C is a valid marker of GFR in human but its relevance, in mice, is Conclusion: Our study shows that the activation of Notch3 is involved in kidney unknown. We prospectively compared cystatin C (PETIA and ELISA), creatinin remodeling in crescentic GN, especially by promoting the migration of podocytes (Jaffe and enzymatic assays) and urea to GFR, measured by inulin clearance in and by activating pro-inflammatory pathways. These results imply that Notch3 male or female C67Bl/6 mice, aged between 16 and 40 weeks, healthy or sub- activation is involved in the physiopathology of CKD, at least in this model, and jected to chronic renal failure by partial bilateral nephrectomy or unilateral suggest that inhibiting the activation of Notch3 could be a novel therapeutic nephrectomy. Analysis of renal function was performed at least 15 days after ini- approach. tial procedure. Keywords: Glomerulonephritis, Notch3, inflammation. A first analysis was performed on 41 mice: urea, enzymatic creatininemia and ELISA cystatin C were significantly correlated to mGFR. Neither PETIA Cystatin C, nor Jaffe creatininemia were correlated with mGFR. We therefore focused on 03-06 ELISA cystatin C, enzymatic creatininemia and urea using complementary An anthropomorphic model of oxalate-dependent whewellite groups. Urea was significantly but weakly correlated to mGFR (r Spearman nephrolithiasis in hypocitraturic rats À0.28, P < 0.05). ELISA Cystatin C and creatinin were significantly correlated F Praddaudea, M Buleona, J Allarda, A Jaafara, M Daudonb, I Tacka aLaboratoire although cystatin C correlation was tighter (r Spearman À0.76 and À0.62, de Physiologie et INSERM UMR1048, CHU Toulouse, Toulouse, France; bService respectively). A Cystatin C-based equation to estimate GFR in mice was devel- d’Explorations Fonctionnelles, Hopital Tenon, APHP, Paris, France oped. In conclusion our results indicate that ELISA cystatin C, but not PETIA, is Whewellite (Wh), the oxalate-dependent form of calcium oxalate (CaOx) lithiasis, better correlated to GFR in mice than enzymatic creatininemia and could be an is difficult to prevent otherwise than by diet. Relevance of the experimental mod- interesting GFR marker. Conversely, the use of Jaffe creatininemia and urea to els of CaOx lithiasis in rodents is controversial since they rely either on adminis- estimate GFR is not relevant. tration of a tubulotoxic oxalate precursor (Ethylene Glycol) or on the Keywords: Glomerular Filtration Rate, mice, cystatin, creatinin, urea, inulin administration of L-Hydroxyproline (HP) plus oxalate that, in fact, do not induce clearance. kidney stones. We assumed that rodents do not develop urinary CaOx stones spontaneously because their high urinary oxalate excretion and concentration are balanced by a high excretion of citrate, a powerful inhibitor of crystallization. 03-04 Therefore, a new model of lithiasis based on hypocitraturia induced by a chronic Tubular STAT3 mediates paracrine tubulo-interstitial communication acid load was developed in rat. during chronic kidney disease progression Twenty adult Sprague-Dawley male rats were divided in two groups: group I F Bienaimea, M Mourahb, M Gallazinnib, S Gabrayc, M Pontoglioc, received a lithogenic diet containing 3% of HP and 3% sodium oxalate and F Terzib aINSERMU845, Centre de recherche Croissance et Signalisation, Equipe 0.21 M NH4Cl in drinking water for 10 weeks whereas group II (controls) ‘mecanismes et startegies therapeutiques des maladie renal chronique’ et service received standard diet and tap water. Urinary parameters (osmolality, citrate, d’exploration fonctionelle, Hôpital Necker, Paris, France; bINSERMU845, Centre de oxalate, urinary acidification and dipstick) were analyzed weekly and creatinine recherche Croissance et Signalisation, Equipe ‘mecanismes et startegies therapeutiques clearance was performed every 3 weeks. After 10 weeks, arterial blood was col- des maladie renal chronique’ Hôpital Necker, Paris, France; cINSERM U1016, CNRS lected, X-ray and photographs of uro-genital apparatus were performed. Stone UMR 8104, Institut Cochin Departement Genetique et Developpement, Universite composition was analysed using infrared spectrometry (IRS). Paris Descartes, Paris, France All rats from group I developed millimetric calculi and four had migrating stones The transcription factor signal transducer and activator of transcription 3 in the ureter while no lithiasis was observed in group II. IRS analysis was typical (STAT3), is suspected to play a detrimental role in the progression of chronic kid- of Wh. Despite a lower body weight, lithiasic rats developed neither renal failure ney disease (CKD) but the molecular basis of its action(s) remains unknown. By nor metabolic acidosis. Treatment resulted in a marked hypocitraturia whereas applying an experimental model of nephron reduction (Nx) to mouse strains with oxaluria was increased. Calciuria remained unchanged. Urine and blood osmolal- different susceptibilities to the development of renal lesions we demonstrate that ity were unaffected. Finally in group I, urine pH was decreased, titrable acidity the degenerative process triggered by nephron reduction is associated with an and ammonuria were increased and haematuria has been present since the third early activation of STAT3 in renal tubules. Specific deletion of Stat3 in renal week (P < 0.05 for all data). tubules dramatically reduced the extent of tubulo-interstitial lesions after Nx. Sur- In conclusion, rats exhibiting a hypocitraturia induced by a chronic acid load prisingly, tubular Stat3 deletion specifically resulted in an important reduction in (NH4Cl) are prone to develop renal stones when exposed to a large amount of the number of activated interstitial fibroblasts, a cell population derived of resi- oxalate. They develop bilateral nephrolithiasis closely mimicking human Wh kid- dent renal pericytes that express the platelet derived growth factor receptor b ney stones. This model sustains the critical role of citrate for the protection of (PDGFRb), suggesting that tubular STAT3 could induce the expression of (a) Wh kidney stones in rat, but also possibly in human. paracrine factor(s) acting on pericytes. Through transcriptomic analysis, compar- Keywords: lithiasis, experimental model, oxalate, rats, citraturia, Whewellite. ative genomics, immunohistochemical and in situ hybridisation studies, we further identified PDGFB and lipocalin 2 as important soluble mediators of STAT3 deleterious effects after Nx. Our study unveils the critical role of tubular STAT3 03-07 in CKD progression and provides important insight into the cross talk between Partial genetic deficiency in tissue kallikrein impairs Human adaptation tubular and interstitial cells that is responsible for kidney lesion progression. to high potassium intake Keywords: Chronic Kidney Disease, STAT3, Tubular cells, Fibrosis, Fibroblasts, A Blancharda, J Monteirob, E Currisc, R Chambreyd, X Jeunemaitrea, PDGFb, Lipocalin2. M Azizia aFacultedemedecine Paris Descartes, Assistance publique des hôpitaux de Paris, Hôpital Europeen George Pompidou, Paris, France; bFaculty of Medicine, Porto University, Porto; cLaboratoire de Biomathematiques, Faculte de Pharmacie, Universite Paris Descartes, EA4466, Sorbonne Paris cite, Paris, France; dINSERM, UMR970, Paris, France Background: Inactivation of the tissue kallikrein (TK) gene in mice impairs renal handling of potassium (K) and favors hyperkalemia. We investigated whether a loss-of-function polymorphism of the human TK gene (R53H) which induces a 50% decrease in enzyme activity would affect renal handling K (1).

Methods: In a crossover study, 30 R53R homozygous and 10 R53H heterozy- tensive patients treated by RAS inhibitors, patients with primary or secondary gous healthy male individuals were randomly assigned to a low-Na/high-K (<20/ hyperaldosteronism, but not in diabetic with elevated Prorenin. 100 mmol/d) diet or a high-Na/low-K (250/40 mmol/d) diet to modulate TK Conclusion: sPRR can be reproducibly measured in human plasma. It is influ- synthesis. On the 7th day of each diet, the participants were studied before and enced by ethnicity, but not by age, sex, hormonal status, circadian cycle or posi- during a 2-h infusion of 20 mg furosemide that functionally excludes the thick tion. The mechanism and significancy of the increased in plasma sPRR in ascending limb and stimulates distal K secretion. patients with primary or secondary hyperaldosteronism or treated by a RAS Results: Plasma K concentration and K renal reabsorption were similar in both inhibitor remain unsettled. The dissociation between (pro)renin ans circulating genotypes on ad libitum Na/K diet or after 7 days of high-Na/low-K diet. Con- sPRR comfort however for a key role of sPRR/PRR in the tissue rather systemic versely, after 7 days of low-Na/high-K diet plasma K was significantly lower in RAS activity. R53R than in R53H subjects [3.64 (3.44; 3.84) vs. 3.85 (3.70; 4.16), respec- Reference: tively P = 0.005] despite similar urine K excretions and plasma aldosterone con- 1. Couisin C. J. Nephrol. (2010) 23 508–513. centrations. On the low-Na/high-K diet, furosemide-induced change in plasma K Keywords: (Pro)renin receptor, diabetes, hypertension, ethnicity, renin, prorenin, was significantly larger in R53H than in R53R subjects [D (K)p: À0.77 mM aldosterone. (À1.12; À0.41) vs. À0.42 mM (À0.52; À0.32), respectively; P = 0.007] showing stimulation of potassium secretion in TK deficient subjects. Conclusions: Impaired TK stimulation by low-Na/high-K diet in R53H subjects 03-10 highlights an inappropriate renal adaptation to K load through an aldosterone Periostin as mediator of fibrotic and inflammatory processes in independent mechanism. Because the defect in renal K excretion occurs in spite crescentic glomerulonephritis of stimulated K secretion, K reabsorption should be even more stimulated than K C Alfieria, P Kavvadasa, JC Dussaulea, C Chatziantonioua aInserm UMR S 702, secretion. These data in TK deficient subjects are in accordance with experimen- Hôpital Tenon 75020 Paris, Paris, France tal data in TK null mice showing stimulation of H,K-ATPase activity (2). Background: We have previously demonstrated the role of Periostin (POSTN), a Reference: matricellular protein, as marker of disease progression in hypertensive nephropa- 1. Slim R., Torremocha F., Moreau T., Pizard A., Hunt S.C., Vuagnat A., Wil- thies. The aim of the present study was to evaluate if the reduction of POSTN liams G.H., Gauthier F., Jeunemaitre X., Alhenc-Gelas F. Loss-of-function poly- expression could affect the degree and the progression of the disease in a model morphism of the human kallikrein gene with reduced urinary kallikrein activity. of crescentic glomerulonephrites. J. Am. Soc. Nephrol. (2002) 13 968–976. Material and methods: Crescentic glomerulonephritis was induced by retro-orb- 2. Chambrey R., Picard N. Role of tissue kallikrein in regulation of tubule func- itary injection of 24 lL/g/body weight of sheep nephrotoxic serum (NTS) in SV/ tion. Curr. Opin. Nephrol. Hypertens. (2011) 20 523–528. 129 mice: seven wild type (WT NTS P), four heterozygous (HE NTS) and six Keywords: Tissue kallikreine, potassium, kidney. knock out (KO NTS) for POSTN. Seven other WT NTS mice were injected daily from D4 to D10 with a specific antisense oligodeoxynucleotide (AS NTS). All the mice were sacrificed 11 days after the last NTS injection. Animals treated with 03-08 scramble oligonucleotide (SCR NTS, n = 7) and WT NTS P were used as disease Metabolic acidosis in homozygous sickle cell disease: prevalence and risk controls. factors in a population of 428 adult patients Results: NTS injection was followed by a rapid increase in POSTN expression in S Maurela, F Lionneta, V Avellinoa, A Girshovichb, K Stankovica, E Letavernierc, WT NTS P and SCR NTS groups. During the follow up, WT NTS P mice showed a R Girotc, JP Haymannc aAPHP, Paris, France; bUMRS 702, Paris, France; cAPHP/ strong disease and high mortality (n = 6). The significant decrease in POSTN UPMC, Paris, France expression observed in AS NTS, HE NTS and KO NTS resulted in a reduction of Introduction: Few reports adress the issue of metabolic acidosis in homozygous fibrotic and inflammatory mediators compared with SCR NTS. When renal tissue sickle cell disease patients which appears as an ammonium synthesis defect was analysed, those groups showed also less deposition of fibrin but, conversely, we responsible for renal tubular acidosis. However, metabolic acidosis prevalence in did not observe any significant protection against glomerulosclerosis, crescents, a non chronic kidney disease (CKD) population is unknown nor acidosis magni- and deterioration of renal function (plasmatic urea and urinary protein excretion). tude or its associated risk factors. Keywords: Periostin, crescentic glomerulonephrites, renal fibrosis, renal inflam- Materials and methods: We performed a cross sectional retrospective study mation. from an adult sickle cell disease (HbSS) cohort of 428 patients followed in a single center. Routine biological parameters were collected and also HbSS clinical complications (such as acute chest syndrome, retinopathy, ulcers…) and ongoing 03-11 medications (such as hydroxyurea and blood transfusions). Metabolic acidosis Targeting connexin 43 protects against the progression of chronic was assessed by plasma bicarbonate concentration. kidney disease in experimental nephropathy in mice Results: Three hundred and ninety patients had an estimated glomerular filtra- a a a a a a 2 A Abed , J Toubas , P Kavvadas , F Authier , C Alfieri , JC Dussaule ,C tion rate (eGFR) using MDRD formula >90 mL/min/1.73 m with a prevalence Chatziantonioua, CE Chadjichristosa aINSERM U702, Paris, France for metabolic acidosis of 26% and 52% in males (16–31 mM) and females (15– Infiltration of inflammatory cells and increased expression of proinflammatory 31 mM), respectively (P < 0.001). Plasma bicarbonate concentration was below factors are crucial events in the development of renal fibrosis during the progres- 20 mM in 7% of cases (n = 27). Metabolic acidosis was associated positively (i) in sion of chronic kidney disease (CKD). We have recently reported that upregula- males with albuminuria independently of eGFR (P < 0.01) (ii) in females with tion of the gap junctional protein Cx43 may be considered as an early signal of age and reticulocyte number. Preliminary data performed in a subgroup suggests CKD. To determine whether Cx43 upregulation is causally related to renal dis- that metabolic acidosis would be due to an ammonium synthesis impairment. ease, we interbred RenTg mice, a genetic model of hypertension-induced CKD, Conclusion: Metabolic acidosis prevalence is high, especially in HbSS female with heterozygous Cx43 to generate RenTgCx43+/À mice. Ten mice per group patients and is not obviously associated with other metabolic disorders. In males, were sacrificed at 5 months and assessed for inflammation and interstitial fibro- metabolic acidosis is associated with albuminuria. Treatment and sickle cell dis- sis. qPCR showed a marked upregulation of cell adhesion molecules (CAM) ease complications were not identified as risk factors. expression such as VCAM-1 (3.5-fold, P < 0.001), and CCR2 (1.9 fold, P < 0.05) Keywords: Sickle cell disease, Metabolic acidosis, Prevalence, Albuminuria. in RenTgCx43 + /+ compared to WT controls. Interestingly, RenTgCx43+/À mice presented lesser upregulation of VCAM-1 (2.1 fold) and CCR2 (1.1 fold) mRNAs (P < 0.05). Consequently, F4-80 immunostainings revealed reduced 03-09 monocyte infiltration in the renal cortex of the RenTgCx43+/À compared to The soluble form of (pro)renin receptor (sPRR): systematic assessment of RenTgCx43 + /+ mice (4.3 Æ 1% and 1.8 Æ 0.2% of cortex surface respectively, circadian variation in plasma of caucasian and non caucasian healthy P < 0.05). In addition, mRNA for type I and III collagen, as well as Sirius Red volunteers and in patients with contrasted activation of the Renin colorations, shown limited renal fibrosis in RenTgCx43+/À mice (0.8% vs. 1.3% Angiotensin System for RenTgCx43+/+, of cortex surface, P < 0.05). Moreover, estimations of histo- A Blancharda, D Bergerota, Y Chambona, E Currisb, G Nguyenc, logical and functional parameters such as albuminuria, glomerulosclerosis and M Azizia aAssistance Publique des Hôpitaux de Paris, Hôpital Europeen George tubular dilation were significantly blunted in RenTgCx43+/À mice confirming a Pompidou, Paris, France; bFaculte de pharmacie, Universite Paris Descartes, Paris, better preservation of the renal function and structure. To investigate whether France; cINSERM Unite Mixte de Recherches S 691, College de France, Paris, France Cx43 could be a potential therapeutic target against the progression of CKD, we Background: A soluble form (sPRR) of the tissue (pro)renin receptor generated by administered a Cx43 oligodeoxynucleotide antisense (ODNAS) in 10 month-old intracellular cleavage by furin and secreted in plasma in animal models and is able RenTg mice, characterized by a more advanced stage of renal disease, during to bind renin and prorenin (1). The determinants of plasma concentration of its solu- 1 month via minipump infusion. In these mice, interstitial renal inflammation ble form, sPRR, is unknown. We thus studied plasma sPRR concentration in normal and fibrosis were blunted and consequently CKD progression was hindered as subjects and patients with contrasted renin angiotensin system (RAS) activation. renal function and structure were highly improved. The same results were Methods: Blood was sampled in 20 healthy male (10 White, 10 Black) in semi- obtained in the Unilateral Ureteral Obstruction model confirming that the protec- recumbent position every 4 h over 24 h and after 1 h standing for plasma levels tive effect of Cx43 blockade is not model-dependent and can be considered as a determination of sPRR, active and total renin, and aldosterone. Hormones were more common protective mechanism of renal disease. These data underline the measured in 120 normotensive healthy subjects (18–75 yrs-old, 60 men/60 importance of gap junctions in renal diseases, and lead to new therapeutic strate- women), 40 patients with diabetes (20 with microangiopathy) 44 hypertensive gies targeting Cx43 to protect against the progression of CKD. patients (22 treated with RAS inhibitors), 11 patients with primary hyperaldoste- Keywords: chronic kidney disease, gap junction, gap junction intercellular com- ronism and 10 with secondary hyperaldosteronism due to a salt losing tubulopa- munication, Connexin 43, cell adhesion molecules. thy. Plasma sPRR was measured using a solid phase sandwich ELISA (IBL International GmbH). Results: Plasma sPRR was 25% higher in White than in Black subjects 03-12 (P < 0.001) whereas plasma total and active renin and aldosterone concentra- Renal stone disease risk factors: influence of a daily green tea intake tions were similar. Within- and between-subject variability in plasma sPRR con- J Rodea, A Dessombesb, L Benzerarac, J Hubert-Brierec, E Letavernierd, M Tliguic, centration was smaller in white (5.0 Æ 1.2% and 11.0 Æ 1.4%) than in Black O Traxerd, M Daudonc, JP Haymannd aUMRS-702, Paris, France; bUPMC, Paris, subjects (9.0 Æ 2.0% and 15.0 Æ 2.3%). It did not display any circadian cycle France; cAP-HP, Paris, France; dUPMC/AP HP, Paris, France and was unaffected by orthostatism, sex, age, hormonal status. It did not corre- Introduction: Renal stone disease prevalence is around 10% in western coun- late with active renine, prorenin or aldosterone plasma concentrations. Plasma tries. Whereas a genetic background is identified in some cases, environmental sPPR concentrations were significantly higher than in control subjects in hyper- factors, mainly food and drinks are well known risk factors. A green tea con-

sumption increase was noticed in a recent survey performed by the CLAFU: 40% To evaluate VD status, one should measure 25OHD, not calcitriol. Although of females aged 40–60 years old are drinking green tea infusions daily. The effect absolute consensus does not exist, most experts consider that, based on muiscul- of green tea in renal stones is debated: it would be a risk factor due to oxalate oskelettal effects of VD, VD deficiency corresponds to a serum 25OHD level below concentration, but would provide protection in rats despite an increased 20 ng/mL (50 nM), while insufficiency corresponds to a 25OHD of 20–29 ng/mL. calciuria. Optimal levels are therefore obtained for values of 30 ng/mL (75 nM) and more. The aim of our study was to evaluate the influence of daily green tea drinks in a Although VD intoxication does not occur with 25OHD levels of <150–200 ng/ renal stone disease population and to study stone morpho constitutional charac- mL, level s of 60–80 ng/mL (150–200 nM) should be currently considered as the teristics. upper optimal levels in the absence of studies showing the safety and/or benefit Material and methods: Four hundred and twenty-three hypercalciuric stone of having higher values in the long-term. formers who underwent a routine evaluation including food regimen inquiry, calcium load, crystalluria and stone morphoconstitutional analysis (when available) were retrospectively selected between 2009 and 2011. Patients with diagnosis of 04-01 primary hyperparathyroidism, sarcoidosis, pregnancy or under biphosphonate, Growth hormone replacement therapy in old rats protects against vitamin D treatment or intermittent green tea consumption were excluded. sarcopenia through antioxidant mechanisms Patients drinking at least 250 mL/day of green tea infusions (n = 48) and con- T Briochea, MC Gomez-Cabreraa, A Gratas-Delamarcheb, J Tresguerresc,J trols defined as non green tea drinkers (n = 239) were compared. Scanning elec- Vina~ a aDepartment of Physiology, University of Valencia, Valencia, Spain; bLaboratory tron microscope (SEM) analysis was performed in 26 stones from the green tea ‘Movement Sport and Health Sciences’, University Rennes 2-ENS Cachan, Rennes, group and 31 stones from controls, all selected for a high weddellite content. The France; cUniversity Complutense of Madrid, Madrid, Spain samples were compared for several parameters including porosity and pore Sarcopenia is defined as the progressive decline of skeletal muscle mass and shapes. strength, which occurs with aging (Hughes et al. 2002; Marzetti and Leeuwen- Results: There was no significant difference between the two groups, both in burgh 2006). Mechanisms that regulate age-related loss of skeletal muscle mass males and females, for demographical, clinical and biological data except a higher are not well defined, but the pathogenesis is likely multifactorial and involves at frequency of hyponatremia in females drinking green tea and of metabolic alcalo- least alteration of the growth hormone (GH)-insulin-like growth factor-1 (IGF-1) sis with a decreased creatinine clearance in males thus suggesting a diuretic axis (Iranmanesh et al. 1991), mitochondrial biogenesis (Dirks et coll. 2006) and effect. Spectrophotoscopy analysis showed a significant higher prevalence of myogenesis (Le Grand and Rudnicki 2007) that are influenced by oxidative stress weddellite in the green tea group. SEM analysis of samples showed a similar fre- (OS). Moreover, it is well established that OS increase with aging in skeletal mus- quency of pores in stones from both groups but a significant predominance of cle (Ji et al. 1990), and this is positively correlated with skeletal muscle atrophy square pores (92% vs. 40% respectively in green tea and control groups, induced by aging (Muller et al. 2007). P < 0.01). The aim of our study was to determine whether administration of GH could pre- Conclusion: Our data suggests that daily green tea intake has a diuretic effect vent the aging-induced alterations in the skeletal muscle of old rats. and do not increase renal stone risk factors. Twenty-four male Wistar rats: 1 months old (n = 8) and 22 months old Keywords: renal stone, green tea, Scanning electron microscopy, diuretics. (n = 16), were divided into three experimental groups: young control (YC, untreated), old control (OC, untreated); and old treated with GH (OGH). Treated animals received physiologically equivalent doses of GH administered for 2 months [2 mg subcutaneously (s.c.)/kg/day]. After treatment, we analysed in SESSION 4: ENDOCRINOLOGIE- the gastrocnemius muscle, mitochondrial biogenesis pathway (PGC-1 alpha and cytochrome C and citrate synthase activity), myogenesis pathways (proteins syn- NEUROENDOCRINOLOGIE thesis through Myostatin, pP70SK6 and heavy myosin chain, satellite cells proliferation through p21 and Myf-5), and oxidative stress parameters (proteins and Treatment of osteoporosis: news and perspectives DNA oxidation and protein levels of antioxidant enzymes). We also analysed IGF- M Audrana, E Legranda, D Chappardb aService de Rhumatologie, GEROM, CHU et 1 plasmatic levels. FacultedeMedecine d’Angers, LUNAM, Angers, France; bLHEA, GEROM, Angers, Aging was associated with an impairment in mitochondrial biogenesis pathway France markers and in myogenesis pathways markers (proteins synthesis as well as Osteoporosis is characterized by low bone mineral bone mass and micro-architec- satellite cells proliferation). At the same time antioxidant defences were decrease tural disruption of the bone tissue. It is a very common disease exposing to exces- and oxidative damages were increase. GH replacement therapy reversed the effect sive bone fragility and increased fracture risk. The fracture-attributable morbidity of aging in the described parameters of old rats. Since sarcopenia is due in part and mortality is especially high after hip and vertebral fractures. Different anti- to an impairment of the mitochondrial biogenesis and the myogenesis, and since osteoclastic and anabolic agents play an important role in the treatment. Their those pathways are negatively influenced by oxidative stress, our results suggest goal is to reduce fracture risk. that growth hormone therapy replacement could protect against sarcopenia Bisphosphonates (administered either orally or intravenously), SERMs, teripara- through antioxidant mechanisms but the exact pathway remains to be deter- tide (rhPTH 1–34), strontium ranelate, in combination with calcium and vitamin mined. D, have demonstrated their usefulness in reducing fracture risk, and are currently Keywords: Growth Hormone, Sarcopenia, Mitochondrial Biogenesis, Myogenesis, used in the prevention of the disease and its complications. [We regret that anti- Oxidative Stress. Rank L (denosumab), whose effectiveness has been proven, used in the USA and Europe for over 3 years, has not been registered in France…] But for several reasons such as a search for a better efficiency and safety, there is 04-02 a need for new bone active agents! Function of osteoblasts and osteoclasts is bet- Muscle insulin resistance precedes the onset of insulin resistance in ter understood and exciting discoveries have been made on the osteocyte. adipose tissue: study in obese postmenopausal women Therefore, the identification of new potential actions on bone cell metabolic path- A Sultana, O Fabreb,CFedouc, K Lambertd, AM Dupuye, JP Cristole, A Avignonf, ways opens exciting new insights: inhibition of bone resorption by cathepsin K J Mercierc, C Bisbalb aCHU Lapeyronie Nutrition-Diabete -INSERM U1046, inhibitors, stimulation of anabolic activity by agents inhibiting sclerostin (and Montpellier, France; bINSERM U1046, Montpellier, France; cCHU Lapeyronie dickoppf-1?) or analogues of PTHrp (calcilytics seem the ‘offside’), potential inter- Physiologie Clinique-INSERM U1046, Montpellier, France; dUMI-INSERM U1046, est of semaphorin 3A due to its dual action on bone resorption and bone forma- Montpellier, France; eCHU Lapeyronie Biochimie, Montpellier, France; fCHU tion… Lapeyronie Nutrition-Diabete-INSERM U1046, Montpellier, France Introduction and objective: Obesity has a strong impact on health by support- ing the development of chronic diseases such as type 2 diabetes. While the majority of obese people develop metabolic complications and insulin resistance, 30% of them are ‘metabolically healthy’ with a level of insulin sensitivity comparable to Vitamin D and osteoporosis those of normal weight, insulin-sensitive persons. To identify the mechanisms lead- JC Souberbiellea aPhysiology Department, Necker Hospital, Paris ing from obesity to insulin-resistance, we analyzed at systemic level and in subcuta- Nutritional sources of vitamin D (VD) are scarce, the only significant ones being neous adipose and muscular tissues, insulin signaling, innate immunity activation, found in fatty fish for VD3, and in some mushrooms for VD2. In fact, VD is not a inflammatory response, oxidative stress, in three groups of subjects: obese insulin- true vitamin as VD3 may be synthesized in the skin under UVB rays exposure. sensitive, obese insulin-resistant and normal-weight subjects. Furthermore, VD needs to be transformed twice to become fully active. A first Patients and methods: Postmenopausal women (50 < age < 65 years): five hydroxylation occurs in the liver to form 25OHD, and a second hydroxylation controls subjects (BMI < 25 kg/m2), 10 obese subjects (BMI > 30 kg/m2): five occurs in the kidney to form calcitriol, the active metabolite. This second hydrox- insulin-sensitive (HOMA < 3) and five insulin-resistant (HOMA > 3) obese sub- ylation is tightly regulated, stimulated by PTH and inhibited by FGF23 and calcit- jects, matched for age and BMI (obese subjects). Systemic characterization riol itself. Calcitriol is released in blood and binds to a receptor, the VDR, in included: hyperinsulinemic-euglycemic clamp, adipokines and cytokines concen- different target tissues. This corresponds to the definition of a true hormone. Cal- trations, and markers of oxidative stress (isoprostanes). Tissue characterization of citriol may be also synthesized in many other tissues where it acts locally muscle (vastus lateralis) and subcutaneous adipose tissue included: insulin through binding to the local VDR. This is the basis for many potential ‘non-clas- response, innate immune response, inflammatory and anti-inflammatory sical’ effects of VD. The main classical biological effects of calcitriol are to stimu- response, oxidative stress and antioxidant defenses measurement. Macrophages late intestinal absorption of calcium and phosphate, promote bone and lymphocytes infiltration in adipose and muscle tissues was also evaluated. mineralization, stimulate bone resorption and calcium reabsorption in the distal Results: In adipose tissue, insulin response is correlated with systemic insulin tubule, and control PTH secretion. Profound VD deficiency cause rickets/osteoma- sensitivity measured by hyperinsulinemic-euglycemic clamp. However, muscle of lacia whereas less severe deficiency may induce a secondary hyperparathyroidism obese insulin-sensitive subjects already presents an insulin resistance, while sys- and increases the risk of falls in the elderly, therefore worsening the risk of osteo- temic insulin sensitivity is maintained. Moreover, adipose tissue and muscle of porosis fractures. Randomized placebo controlled trials of VD, usually with cal- obese insulin resistant subjects show an activation of the inflammatory, anti- cium, have shown that approximately 800 IU/day of VD significantly decreased inflammatory and anti-oxidant response. the risk of non vertebral fracture in osteoporosis patients and of falls in the Conclusion: In humans, muscle is more sensitive to the deleterious effects of elderly. Studies have also shown that most osteoporosis treatments are less effec- inflammation and oxidative stress. Insulin resistance seems to first develop in tive when administered to VD insufficient patients. In many countries, evaluation muscle before appearing in adipose tissue, although it also presents a micro- of VD status and supplementation in case of insufficiency has become a prerequi- inflammation. site to the prescription of osteoporosis treatment. Keywords: obesity, insulino-sensitivity, adipose tissue, muscle, inflammation.

04-03 04-05 Increased production of free radicals: a new pathogenic role for Metabolic syndrome and urinary lithiasis mitochondria in inflammatory myopathies? D Hadj Merabeta, K Bereksi Reguigb aBiology Department, Sidi Bel Abbes; A Meyera, M Hengenb, AL Charlesb, J Zollb, F Singhb, AI Schlagowskib,A bLaboratory of Environment Synthesis, Sidi Bel Abbes Echanizc, J Sibiliad, JE Gottenbergd, G Bernarde aExploration Fonctionnelle Objectives: Closer the links between these two pathologies that have alarmingly Musculaire – Mitochondrie, Stress Oxydant et Protection Musculaire EA 3072, propagated into the entire world without exception by assessing the epidemiologi- Strasbourg, France; bMitochondrie, Stress Oxydant Et Protection Musculaire EA cal parameters. 3072, Strasbourg, France; cService de Neurologie – Hôpitaux Universitaires de Patients and methods: An epidemiological investigation was conducted in the Strasbourg, Strasbourg, France; dService de Rhumatologie Centre de refgerence des city of Sidi Bel Abbes (Algeria). Our study was performed on patient files at the maladies autoimmunes rares – Hôpitaux Universitaires de Strasbourg, Strasbourg, Endocrinology Service during 3 years [(12/2006–11/2007)(12/2007/11/2008) France; eService des Explorations Fonctionnelles-, Strasbourg, France (12/2008–11/2009)]. Introduction: Non-immunological mechanisms are emerging players in the The methodology of the statistical analysis was based in two axes: 1- Descriptive pathophysiology of inflammatory myopathies (IM) (Henriques-Pons et al. 2009). statistics (Prevalence & Incidence) and 2- Analytic statistics (Exposed /Non Surprisingly, oxidative stress in the muscle of IM patients has not been explored. Exposed: relative risk). H2O2, which is mainly produced by NADPH oxidase, xanthine oxidase and mito- Results and discussion: The epidemiological study of 600 files shows that the chondrial respiratory chain, can play a direct toxic effect on muscle functioning prevalence of Metabolic Syndrome (MS) [NCEP ATP III, 2002] is 42.66% which (Westerblad 2011) and act as a chemokine for inflammatory cells (Niethammer is 1.7 times higher among women compared to men and the prevalence of Uri- et al. 2009). We determined whether H2O2 level is increased in muscle of IM nary Lithiasis (UL) in the general population is 6.16% whereas in those with MS patients and investigated whether mitochondria participate in H2O2 production. is 11.32%. We also noted an increase in the incidence of MS and its risk factors Methods: Five patients with IM were included in this study. Five patients suffer- parallel to that of the UL. For 2006–2007: MS (43.47%), UL on all population ing from myalgia, but without muscle pathology, were used as controls. Samples (3.91%), UL on those with MS (0.3%). For 2007–2008: MS (39.25%), UL on all from deltoid muscle were obtained at the time of muscle biopsy performed for population (6.07%), UL on those with MS (13.09%). For 2008–2009: MS diagnosis purpose. The production of H2O2 was recorded in the presence of dif- (46.15%), UL on all population (9.61%), UL on those with MS (20.83%). The ferent substrates increasing the activity of the mitochondrial respiratory chain results show also that the RR is >1 among patients who have the MS than those using spectrofluorometry (Amplex red). unscathed and this proves that MS is a risk factor of UL. Results: Average age was comparable in both groups (IM: 44.4 Æ 13.2 years; Keywords: Metabolic Syndrome, Urinary Lithiasis, Risk Factor, Epidemiology, controls: 44.2 Æ 10.7 years). The sex ratio was ¼ in both groups. Prevalence, Incidence. Without mitochondrial substrates, H2O2 production was similar in both groups (IM: 1.94 Æ 1.44 pmol/min/mg; controls: 1.21 Æ 0.75 pmol/min/mg, P = 0.55). After addition of glutamate and malate, H2O2 production was higher in IM group (5.55 Æ 3.45 vs. 1.06 Æ 1.03 pmol/min/mg, P < 0.04). Similar SESSION 5: A3P COMMUNICATIONS LIBRES results were obtained with succinate (11.5 Æ 5.22 vs. 2.54 Æ 2.26 pmol/min/ mg P < 0.04) and ADP (7.38 Æ 6.84 vs. 0.52 Æ 0.21 pmol/min/mg P < 0.04) (Figure 1). Management of bleeding in patients taking new oral anticoagulant – treatment Discussion Conclusion: These results are the first data available on oxidative a,b a b stress production in muscle of IM patients. Very interestingly, increased H2O2 PMRoy FacultedeMedecine, L’UNAM Universite, Angers, France; Departement production was higher in IM patients as compared with controls only in the pres- de Medecine d’Urgence, CHU Angers, France ence of mitochondrial substrates. This suggests a key role for respiratory chains Bleeding is the major complication of oral anticoagulation therapy and is yet the complexes in H2O2 production during IM. Whether mitochondria from muscle most frequent of major adverse drugs events in terms of morbidity and mortality. fibers or inflammatory cells (or both) are involved in this process require further Vitamine K antagonists (VKA) were the only oral anticoagulants for decades and investigations. are yet the widely prescribed. In recent years, new oral anticoagulants (NOACs) References: that selectively inhibit either thrombin (dabigatran etexilate) or factor Xa (riva- 1. Enriques-Pons et al. Curr. Opin. Rheumatol. (2009) 21 581–587. roxaban, apixaban) have been marketed. They differ in many respects and espe- 2. Westerblad et al. Antioxid Redox Signal. (2011) 15 2487–2499. cially in pharmacokinetics but all have the advantage of having a wide 3. Niethammer et al. Nature (2009) 459 996–9. therapeutic range without requiring regular monitoring. To date, clinical studies Keywords: Inflammatory myopathies; oxidative stress; mitochondria. show that the incidence of spontaneous bleeding with NOACs is comparable to that of established anticoagulants. However, unlike vitamin K antagonists, there are currently no clinically available antidotes or approved reversal agents for NO- 04-04 ACs. For rivaroxaban, a specific decoy is under evaluation (FXa-GLAless), Protective effect of Eucalyptus globulus against acetaminophen induced whereas, for dabigatran, the Company is developing a specific selective antidote. liver damage in male rat Attempts have been made to restore normal coagulation after NOACs using com- S Dhibia, S Mbarkia, A Elfkia, N Hfaeidhb aLabortaoire d’ecophsiologie animale de pounds such as recombinant activated factor VII (rFVIIa), prothrombin complex Sfax, Gafsa, Tunisia; bLabortaoire d’ecophsiologie animale de Sfax, Sfax, Tunisia concentrate (PCC), or FEIBA (factor eight inhibitor bypassing activity). Dabiga- Under our experimental conditions, acetaminophen poisoning resulted in an oxi- tran has the additional option of removal from the blood system via dialysis. dative stress evidenced by a significant increase of lipids peroxidation level in Emergency dialysis can remove about 60% of the dabigatran concentration in hepatic tissues, hyperglycaemia and deccreased levels of total cholesterol and tri- 4h. glycerides, and transaminases activities in blood. Previous administration of Euca- However very limited pre-clinical data and even less clinical evidence are avail- lyptus globulus extract was found to alleviate this damages. able on the usefulness of these methods for the emergency management of criti- Experimental design: Rats were divided into four batches: (C) was the control cal bleedings. So, only weak propositions can be currently performed for group; (P) was a group of rats treated with acetaminophen (300 mg/kg); (P) was management of bleeding events with NOACs in clinical practice, based on old a group of rats given Eucalyptus globulus extract (20 g/100 mL) during 4 weeks; established measures that are the same for all anticoagulant treatments and on (PE) was a group treated by Eucalyptus globulus then injected by acetaminophen some more specific but no validated options. (300 mg/kg). After 4 weeks, animals from each group were rapidly sacrificed. Biochemical assays: Level of lipids peroxidation was measured as thiobarbituric acid reactive substances (TBARS), according to The total (Cu-Zn and Mn) super- 05-01 oxide-dismutase (SOD) activity was determined by measuring its ability to inhibit Increase in the rate of spontaneous adverse drug reaction notification and improvement of pharmacologic quality analysis in a department of the photoreduction of nitroblue tetrazolium (NBT) . One unit of SOD represents â the amount inhibiting the photoreduction of NBT by 50%. The activity was infectious diseases using an electronic medical record: NADIS ° AL Ruellana, C Allavenab, T Jovelinc, E Billaudb, B Bonnetb, N Feuilleboisb, expressed as units/mg protein, at 25 C. Glutathione-peroxidase (GPX) activity b b a a was expressed as lmoles of GSH oxidized/min/g protein. The level of glycaemia, E Lousteau , F Raffi , P Jolliet CHU Nantes – Service de Pharmacologie Clinique – Centre Regional de Pharmacovigilance, Nantes cedex, France; bCHU Nantes – Service cholesterol, triglycerides, and transaminases in serum were determined by kit c methods (Spinreact). des Maladies Infectieuses et Tropicales, Nantes cedex, France; COREVIH Pays de Results: TBARS levels in hepatic tissue were increased in acetaminophen treated Loire, Nantes cedex, France Background: Adverse Drug Reactions (ADRs) are commonly under-reported. rats as compared to controls. Administration of Eucalyptus globulus significantly reduced these TBARS levels. Activities of enzymes that protect against oxidative NADISO is an electronic medical record (EMR) for Human Immunodeficiency stresses, SOD, CAT and GPX were found to be respectively reduced in liver of Virus (HIV) infected subjects seeking care in french public hospitals. An auto- acetaminophen treated rats, as compared to controls. These changes, revealing a matic warning, informing the physician of each new Adverse Drug Reactions (ADRs) associated with Anti-Retroviral Drugs (ARDs) leading to discontinuation, failing defence against an oxidative stress, were largely corrected in animals trea- â ted by Eucalyptus globulus extract. Acetaminophen treatment induced severe liver has recently been added in NADIS . Material and methods: The aim of this study was to measure the quality and damages evidenced in serum by a significant increase of hepaic enzymes. When â acetaminophen treated rats were also treated with Eucalyptus globulus extract, all the benefit of NADIS in the Regional Pharmacovigilance Center of Nantes. ADRs reported via NADISâ pharmacovigilance tool were collected during 21 months theses biomarkers were restored. â References: (from January 2011, start date of NADIS pharmacovigilance tool utilization, to 1. Abdel-Zaher A., Abdel-Rahman M., Hafez M., Omran F. Role of nitric oxide October 2012). Each effect was sent to the Regional Center. After sorting the and reduced glutathione in the protective effects of aminoguanidine, gadolinium data according to System Organ Class (SOC), the rate of ADRs with each drug chloride and oleanolic acid against acetaminophen-induced hepatic and renal was calculated. Severity, evolution, and patient characteristics were also notified damage. Toxicology (2007) and analysed. 2. Clement Y.N., Williams A.F. Protection against paracetamol-induced hepatic Results: Eighty-eight cases were retained for analysis, and observed in 81 HIV- – infected patients. In comparison with a 21 months long lasting period before NA- injury by prazosin pre-treatment in CD-1 mice. Mutat. Res. 579 (2005) 182 â 188. DIS pharmacovigilance tool utilization (from April 2009 to December 2010), an Keywords: acetaminophen Eucalyptus globulus, hyperglycaemia. increase of 60% in collected cases was observed. The mean age of patients was 44 years (SD: 14; min: 18; max: 82), with a sex ratio of 1.79 (M/F). Seven (7.9 %) ADRs were recorded as serious, leading to hospitalisation, or being life threat- ening. The outcome of thirty-nine (44.3%) ADRs were favourable after drug dis-

continuation. Renal and urinary (19.3%), gastro-intestinal (14.8%), nervous sys- later at the age of 6–12 months. No unexpected adverse outcome was observed tem and psychiatric (15.1%), skin and subcutaneous tissue (10.2%), metabolism among the 29 other babies followed up until the age of 1.5–30 months (median: and nutrition (10.2%) disorders were the most frequently described ADRs. Regi- 9.5 months) and nine experienced common infectious episodes. mens including Nucleoside Reverse Transcriptase Inhibitor combined with Non Discussion: Except one debatable case of asymptomatic neutropenia, no biologi- Nucleoside Reverse Transcriptase Inhibitors or with Protease Inhibitors have been cal or clinical consequences of maternal azathioprine treatment were observed in associated with ADRs in 36.4% and 43.2% of cases, respectively. breastfed babies, even after long-term follow-up, or in premature newborn. Discussion/Conclusion: Since the availability of NADISâ pharmacovigilance Although these results are reassuring, infant blood cell counting should be sys- tool, a significant increase in spontaneous notifications has been observed limit- tematically proposed 10–15 days after breastfeeding initiation. ing the under-reporting rate. Pharmacologists and Physicians collaboration asso- Keywords: Azathioprine, Breastfeeding, Long-term follow-up. ciated with suitable information collect via NADISâ can contribute to improve the quality of pharmacologic analysis and the medical following of each report, in order to carry on the benefit/risk ratio regimens. 05-04 Keywords: Pharmacovigilance, Electronic medical record, Anti-retroviral drugs, Evolution and characteristics of pristinamycin-induced adverse effects: Adverse Drug Reactions. analysis from the French National Pharmacovigilance database A Borgnona, L Moachona, ADCR De Pharmacovigilancea aCentre regional de Pharmacovigilance – Hôpital Cochin, Paris, France 05-02 Pristinamycin is the mixture of pristinamycin IA (group B streptogramin) and Adverse drug reactions and off-label drug prescription in paediatric pristinamycin IB (macrolide), active on staphylococci and streptococci. It is avail- outpatients: a survey among general practitioners in south western able in France for over 30 years. Following the observations in our hospital of France adverse effects (AE) unusual for their severity and/or onset delay, our objective R Bissuela, A Palmarob, N Renaudb, G Durrieub, B Escourroua, JL Montastrucb,M was to evaluate pristinamycin-induced AE, qualitatively and quantitatively, along Lapeyre Mestreb aDepartement Universitaire de Medecine Generale des Facultes de the time, from the French National Pharmacovigilance database. Medecine de Toulouse, Toulouse, France; bLaboratoire de Pharmacologie Medicale et Methods: An analysis from the database was performed since 1999, counting Clinique, Centre Midi-Pyrenees de PharmacoVigilance, de Pharmacoepidemiologie et the number of severe and non severe AE, skin and non-skin AE, with pristinamy- d’Informations sur le Medicament, Equipe de Pharmacoepidemiologie, INSERM cin suspected. A more precise analysis compared all the severe AE notified in U1027, FacultedeMedecin, Centre Hospitalier Universitaire, Toulouse, France 2004–2005–2006 to those in 2009–2010–2011. Sails volume was kindly pro- Aim: To study the association between off-label drug prescription and Adverse vided by ANSM. Some cases were reclassified as DRESS syndrome when Kardaun Drug Reaction (ADR) occurrence in a sample of paediatric outpatients managed score was found 3. by general practitioners Results: The number of AE, around 125/year, sharply increased in 2007–2008, Methods: A prospective pharmacovigilance survey on paediatric drug prescrib- to reach 180/year thereafter. The ratio of severe/non severe AE increased from a ing was implemented in a sample of general practitioners from 46 practices in mean of 1.1 before 2007 to 1.8 after 2007. The number of non-severe AE was Midi-Pyrenees area (South western France). All consecutive patients aged 0–16 stable, as well as the proportion of skin AE (severe and not severe) (68.8 % of were included. Characteristics of the patients (age, gender, weight), main reasons total AE). for consulting, and drug prescribed (including strength, route and indication) Comparison of severe AE between the two groups of 3 years indicated that the were collected. Incidence and characteristics of ADRs occurring within 10 days number of digestive AE, hematological AE, bullous dermatitis, DRESS, vascular after the date of consult and identified by the GP were recorded. Off-label pre- purpura, and other skin reactions, had practically doubled whereas sails volume scription was defined as prescribing outside the specifications of the Summary of increased by 35 %. Their median delay of onset was shorter in the second period, Products Characteristics. Unlicensed drugs were those with no valid marketing with a statistically significant difference reached for type I reactions (2 vs. 0.5 h). authorization. Characteristics of age and sex remained similar. In 11 cases of bullous reactions Results: Among the 2313 visits included from March 8, 2011 to July 31, 2011, (including Lyell syndrome) and 23 cases of DRESS, 1 or 2 days only separated 1960 involved at least one prescribed drug with a total of 4888 drugs prescribed. pristinamycin introduction to the reaction onset. Mean age was 5.59 (SD 4.49), with a sex ratio of 1.06. Among the 1960 chil- Discussion: Characteristics of some severe AE are not presently indicated in the dren receiving at least one prescription, 37.6 % (n = 737) were exposed to at summary of product characteristics: mainly cytopenias, and DRESS syndrome least one off-label prescribing, and 6.7% (n = 132) to at least one unlicensed with the possibility of very unusual short delay of occurrence. The increase num- drug. Off-label prescribing involved an unapproved indication in 56.4% of the ber of hypersensitivity reactions together with the short time of onset for DRESS cases (n = 416), a lower dosage than specified in 26.5% (n = 195), a higher dos- and bullous dermatitis suggest that the population has been sensitized to pristina- age than specified in 19.5% (n = 144), age not labeled in 7.2% (n = 53), incor- mycin. A food origin for this sensitization cannot be excluded. rect route of administration in 3.5% (n = 26), and contraindication in 0.3 % Keywords: pristinamycin, severe adverse reactions, French pharmacovigilance (n = 2). A total of 23 ADRs were reported, giving an incidence of 1.0% in the database. population study and of 1.5% in patients exposed to at least one off-label prescription. None of the ADRs was considered as ‘serious’, and all were reported to the pharmacovigilance center. ADRs occurrence was not found to be significantly related to off-label drug prescribing. SESSION 6: PHARMACO CLINIQUE Discussion: Despite the numerous initiatives implemented since 2005 for promoting rational medicines use in children, the magnitude of off-label prescription in outpatient paediatric practice remains high (around 35% to 40%). By contrast 06-01 with the findings of a previous study conducted more than 10 years ago [1], we Pharmacological treatment in type II diabetes and evidence based did not found any increased ADR risk related to off-label prescribing. medicine Reference: R Boussageona, F Gueyffierb,c,d, T Bejan-Angoulvante,f,g, C Cornuh,i aDepartment of 1. Horen B., Montastruc J.L., Lapeyre-Mestre M. Adverse drug reactions and off- b – General Practice, Universite de Poitiers, Poitiers, France; Department of Clinical label drug use in paediatric outpatients. Br. J. Clin. Pharmacol. (2002) 54 665 Pharmacology, Hospices Civils de Lyon, Lyon, France; cUMR 5558 CNRS, 670. Villeurbanne, France; dUniversite Claude Bernard Lyon1, Lyon, France; eDepartment of Keywords: Adverse drug reactions, Off-label drug use/prescription, Paediatric Clinical Pharmacology, CHRU de Tours, Tours, France; fUMR 7292 CNRS, Tours, outpatients, Pharmacovigilance. France; gUniversity Francois Rabelais, Tours, Tours Cedex 9, France; hDepartment of Clinical Pharmacology, Hospices Civils de Lyon, France; iInvestigation Centre, Louis Pradel Hospital, Bron, Lyon, France 05-03 Recently ADA and EASD published a position statement in the management of Azathioprine and breastfeeding: long-term follow-up hyperglycaemia in type II diabetes (T2D), without questioning the ‘treat to target’ N Bernarda, A Gourauda, N Pareta, J Cottina, J Descotesa, T Vialb aCentre Regional dogma, proposing ‘individualized’ HbA1C targets depending on history of diabe- de Pharmacovigilance, Hospices Civils de Lyon, Lyon, France; bCentre Regional de tes, contra-indications and co-morbid conditions. But how strong is the evidence Pharmacovigilance, Hospices Civils de Lyon, Lyon Cedex 03, France for an intensive strategy compared with a less stringent standard? A meta-analy- Objective: Although a number of reports indicate that azathioprine can be safely sis of 13 trials compared the effect of intensive vs. standard treatment in T2D on used during lactation, data on long-term follow-up of breastfed babies are still both microvascular and cardiovascular complications. Intensified treatment did limited. Our aim was to report the biological and long-term clinical follow-up of not reduce all-cause or cardiovascular mortality; a reduction in the risk of non- breastfed infants from azathioprine-treated mothers. fatal myocardial infarction and microalbuminuria was observed, but these reduc- Methods: All azathioprine-treated mothers who contacted Lyon Pharmacovigi- tions were not anymore significant when high quality studies only were consid- lance Center before initiating breastfeeding, and finally opted to breastfeed their ered. The risk of severe hypoglycemia was more than twofold increased, leaving baby for at least 1 month during treatment were included. The follow-up was the benefit-risk ratio uncertain. performed through regular telephone interviews of the mother and/or her pedia- Here we further investigated on the same data the relationship between HbA1c trician. The data collected for the breastfed infants included results of the blood decrease and clinical outcomes as log(OR) in a meta-regression with random cell counts (usually at birth and after 2–3 weeks of breastfeeding), and data on effect model. On average, the decrease in HbA1c was associated with a non-sig- growth and general health. nificant increase in mortality (seven studies), cardiovascular mortality (eight Results: Twenty-nine women (30 newborns) receiving azathioprine 50–175 mg/ studies), and stroke (seven studies). It did not display statistically significant rela- day were included. Eleven babies were born prematurely (27–36 weeks), includ- tion with the decrease in the risk of myocardial infarction (six studies) and heart ing one set of twins. Breastfeeding duration ranged from 1 to 17 months (med- failure (eight studies). The relations between HbA1c and the risk of photocoagu- ian: 3.5 months). Blood cell counts performed at birth in 16 newborns evidenced lation (three studies), visual loss (three studies), worsening of renal failure or a decrease in white cell count in 3, which further normalized despite breastfeed- doubling of plasma creatinin (four studies), neuropathy and peripheral vascular ing continuation, suggesting this might be the consequence of in utero exposure disease (six studies for both) were also inconclusive. to azathioprine. Among the 20 babies who had late blood cell counts, only one This systematic review did not show significant association between HbA1c presented with asymptomatic neutropenia on day 15, but azathioprine metabo- reduction and any clinical benefit. The efficacy of glucose lowering treatments lites below the limit of quantification. Neutropenia fluctuated from 0.5 to 1 G/L has been seriously challenged by meta-analyses, these drugs should be used cau- during the 1.5-month period of breastfeeding, persisted for 15 days after breast- tiously to avoid acute metabolic accidents, e.g. hypoglycemia, and major hyper- feeding discontinuation, and completely resolved 3.5 months later. No other glycemia. The prevention of cardiovascular complications should first rely upon causes were found. Several episodes of non serious winter infections were noted ACE inhibitors and statins that present a much higher level of evidence in that

indication. Recent recommendations for pharmacological treatment in T2D are older than those of female GPs (78.2 vs. 76.9 years), are more often men (30% not supported by evidence from randomized controlled trials. This urges the need vs. 19%) and suffer less frequently from inflammatory pain (25% vs. 33%). for evaluating the clinical benefit of glucose defined by clinical outcomes rather Conclusion: The prescriptions for older patients suffering from chronic pain dif- than glycated haemoglobin as primary endpoint. fer between male and female GPs. However, differences in their patients’ profiles Keywords: type-2 diabetes; intensive glucose lowering treatment; meta-analysis; may be the cause. meta-regression; cardiovascular events; microvascular complication. Keywords: Analgesics, Chronic pain, GPs.

06-02 06-04 Experimental designs for small randomised clinical trials: an algorithm Consistency of safety and efficacy of new oral anticoagulants among for choice patients with comorbidities in non-valvular atrial fibrillation: a C Cornua, B Kassa€ıa, R Fishb, C Chironc, C Albertid, R Guerrinie, A Rosatie, meta-analytic approach G Ponsf, S Chabaudg, D Caudrih, C Balloti, P Kurbatovaj, C Castellank, A Barjardg, JC Legaa, L Bertolletib,c, C Gremilletd,e, C Chapelleb,f, P Mismettib,g, M Cucherath, P Nonyj aUMR 5558, CRNS Lyon- CHU Lyon-Centre d’Investigation Clinique Lyon, S Laporteb,i aHospices Civils de Lyon – Universite Lyon 1 Claude Bernard, Pierre- University of Lyon 1, Lyon, France; bNovartis Pharma, Basel, Switzerland; cINSERM Benite, France; bEA3065, Universite Jean Monnet, Saint-Etienne, France; cUnitede U663 «Epilepsy in childhood and brain plasticity », Hopital Necker – Enfants Malades, recherche Clinique, Innovation, Pharmacologie, Saint-Etienne, France; dA3065, Paris, France; dHopital Robert Debre -Unite d’Epidemiologie Clinique -Inserm CIE-5- Universite Jean Monnet, Saint-Etienne, France; eService de Medecine et Therapeutique, Antenne URC Paris, Paris, France; ePediatric Neurology Unit and Laboratories, CHU Saint-Etienne, Saint-Etienne, France; fUnite de recherche Clinique, Innovation, Children’s Hospital A. Meyer-University of Florence, Florence, Italy; fInserm UMR Pharmacologie, CHU Saint-Etienne, Saint-Etienne, France; gUnite de recherche 663, Paediatric Committee -EMA (London UK), Head PIP WP (AFSSAPS), Cochin – Clinique, Innovation, Pharmacologie et Service de Medecine et Therapeutique, CHU de Saint Vincent de Paul Hospital, University Paris Descartes, Paris, Paris, France; Saint-Etienne, Saint-Etienne, France; hUmr Cnrs 5558 Evaluation et Modelisation des gUnite de Biostatistique et d’Evaluation des Therapeutiques, Centre Leon Berard, Lyon, Effets Therapeutiques, Universite Claude Bernard Lyon 1, Lyon, France France; hErasmus University Medical Center, Sophia Children’s Hospital, Rotterdam, Background: Because of well-known limitations of VKA, new oral anticoagu- The Netherlands; iUMR 5558, CRNS Lyon, University of Lyon 1, Lyon, France; lants (NOAs) inhibiting directly thrombin or activated Factor X have been devel- jUMR 5558, CRNS Lyon- CHU Lyon, University of Lyon 1, Lyon, France; kUMR oped in non-valvular atrial fibrillation (AF) patients. The consistency of their 5558, University of Lyon 1, Lyon Cedex 08, France safety and efficacy profiles are still debated among subgroups of patients with Objectives: Rare diseases are defined on the basis of their low prevalence, i.e. <1 morbidities. in 2000 people and it is estimated that there are between 6000 and 8000 rare Methods: We searched MEDLINE, EMBASE, and abstracts and conference pro- diseases. Whereas the available population of potential research participants does ceedings (up to October 2012) for randomized trials that compared NOAs and not allow the conduct of adequately powered parallel group randomized clinical VKA in AF patients with age over 75 or younger, with/without renal impairment trials (RCTs), several alternative trial designs have been proposed in such a con- or heart failure (HF), VKA experienced or na€ıve patient, CHADS2 score (0–1, 2, text. In order to develop an algorithm to select the most appropriate design(s) for or 3), with/without prior transient ischemic attack or stroke (TIA-S). Main effi- given disease-drug-outcome situations, we performed a literature review of alter- cacy and safety outcomes were stroke and systemic embolism (SSE) and major native trial designs for small clinical trials. bleeding (ISTH guidelines) (MB), respectively. Interaction was assessed across sub- Methods: Literature was searched for trial designs dedicated to randomised, groups with a threshold at P < 0.10. comparative small clinical trials. The characteristics, advantages and limits of Results: Four studies (51867 patients) comparing warfarin to apixaban (1), riva- each identified design were then summarised and an algorithm was developed to roxaban (2), and dabigatran (1) were included. Overall, NOAs were associated select the most appropriate trial design(s) for a given disease / treatment. Decision with a significant reduction in SSE (RR 0.84; CI 0.76–0.92) and MB (RR 0.85; nodes included the characteristics of the clinical outcome (reversible or not), the CI 0.79–0.91). For SSE, No interaction was retained whatever CHADS2 score, delay for the treatment response (fast: weeks; slow: months), whether time under renal insufficiency, age, or prior TIA are. A quantitative interaction was found placebo needs to be minimised, whether all patients must receive active treatment between HF patients (RR 0.90 CI 0.78–1.04, and patients without HF (RR 0.76 before the end of the trial, and whether comparisons should be intra- or inter- CI 0.68–0.86, P = 0.09 for interaction). Less MB were recorded with NOAS even patient. in patients with renal impairment estimated by a Cockcroft-Gault creatinin clear- Results: Twelve trial designs were identified: parallel group; cross-over; latin ance between 30 and 50 mL/min (RR 0.80 CI 0.61–1.06). A significant quanti- square; factorial design; N-of-1; delayed start placebo-phase; stepped wedge; tative interaction (P = 0.02) was found between the subgroup of VKA na€ıve randomised withdrawal; early escape; three-stage; play the winner; drop the patients (RR 0.67 CI 0.64–0.72) compared to the subgroup of VKA-experienced loser. Based on the characteristics of these designs we built a sequential decision subjects (RR 0.78 CI 0.70–0.86). There was no interaction for other outcomes. tree for the choice of the most appropriate design according to each decision node Discussion: The treatment effect of NOA appeared to be better and safer than and illustrated its use through examples of published trials. VKA by reducing SSE and MB whatever the comorbidities of patients. Notably, Discussion: However, many additional factors, such as objective(s) of the trial, moderate renal impairment or high CHADS2 score were not associated with number of patients needed, length of trial, and how the measure variability is more hemorrhagic or ischemic events. Mortality data are not available by sub- managed, are important in the choice of the most suitable study design. Beyond group to date. To conclude, in regards of the limitations of our study, based on our algorithm, the respective performances of these different trial designs have to meta-analytic combination of subgroups, dedicated evaluation of NOAs in tar- be further assessed ‘in silico’ using mathematical modeling and simulation. This geted population are required. will be performed in the ‘CRESIM’ project, founded by the ERANET-PRIOMED- Keywords: apixaban, rivaroxaban, dabigatran, renal failure, CHADS2 score, CHILD European call. heart failure, stroke. Keywords: randomized clinial trial; rare disease, orphan drug.

06-05 06-03 Cutaneous iontophoresis of treprostinil: a pharmacokinetic and Do male and female general practitioners (GPs) prescribe differently the pharmacodynamic study analgesics in the elderly patients suffering from chronic pain? M Roustita, F Gaillard-Bigota, S Blaiseb, F Stanke-Labesquec, C Cracowskid, A Lazkania, T Delespierrea, L Becquemonta, L Benattar_Zibib, P Bertinc, JF Jourdile, B Imbertf, C Seinturierf, P Carpentierb, JL Cracowskia aUnitede G Berrutd, E Corrublee, N Danchinf, G Derumeauxg, B Falissardh, F Forettei, Pharmacologie Clinique, Centre d’Investigation Clinique – Inserm CIC003 & Universite F Pasquierj, M Pingetk, R Ourabahl aPharmacology Department, Faculty of medicine Joseph Fourier, Grenoble, France; bUnitedeMedecine Vasculaire, CHU de Grenoble & Paris-Sud, Paris, France; bORPEA/CLINEA, Puteaux, France; cDepartment of Universite Joseph Fourier, Grenoble, France; cLaboratoire de Pharmacologie et Rheumatology, Limoges, France; dPôle de gerontologie clinique, Nantes, France; Toxicologie, CHU de Grenoble & Universite Joseph Fourier, Grenoble, France; dUnitede eINSERM U 669, Faculty of Medicine Paris-Sud Department of Psychiatry, Paris-Sud Pharmacologie Clinique, Centre d’Investigation Clinique – Inserm CIC003, Grenoble, University, Paris, France; f HEGP, Service des maladies coronaires, Paris, France; France; eLaboratoire de Pharmacologie et Toxicologie, CHU de Grenoble, Grenoble, gExploration Fonctionnelles Cardiovasculaires, Louis Pradel Hospital, Lyon, France; France; fUnitedeMedecine Vasculaire, CHU de Grenoble, Grenoble, France hINSERM U 669, Faculty of medicine Paris-Sud Department of Biostatistics, Paris- Introduction: Systemic sclerosis (SSc) is a rare disease affecting the skin micro- Sud University, Paris, France; iFondation Nationale de Gerontologie, Universite Rene circulation. Therapy of SSc-related ulcers, a severe complication of SSc-vasculopa- Descartes, Paris, France; jUDSL, EA 1046, CHU, Universite Lille Nord de France, thy, relies on IV iloprost. However, therapeutic effects are counterbalanced by Lille, France; kService d’Endocrinologie, Diabete, Maladies de la Nutrition (Pole^ systemic vasodilatation-induced adverse effects. Cutaneous iontophoresis of tre- NUDE), Hopitaux^ Universitaires de Strasbourg et Centre europeen d’etude du Dia- prostinil, another prostacyclin analogue, induces sustained vasodilation on the bete (CeeD), Strasbourg, France; lGeneral Practice Departement, Faculty of medicine forearm of healthy subjects [1]. The objectives of this work are 1/to assess intra- Paris-Sud, Paris, France dermal vs. plasma concentrations of iontophoretically-administered treprostinil on Objective: The aim of this work is to study the influence of GP’s sex on the pre- the forearm of healthy subjects; 2/to assess the effect on the finger pad of healthy scription of analgesics in patients older than 65 years. subjects and perform a dose-finding study; and 3/to test the most appropriate Patients and methods: This is a prospective observational cohort study of dose in SSc patients. 1379 patients over 65 years, living at home, suffering from chronic pain for at Methods: In the first study, healthy participants simultaneously received ionto- least 3 months and requiring care. The patients are followed by GPs installed in phoresis of treprostinil and NaCl on the forearm. Two microdialysis fibers were cabinet. This is one of the three cohorts in the study (S. AGE, aged subjects). GPs inserted at the dermis-hypodermis junction under the treprostinil site, and sam- collected via eCRF all their prescriptions and clinical data of patients. The data ples were collected every hour during 8 h. Blood samples were taken on the con- presented in this study is the baseline data; follow-up of 3 years is underway. trolateral forearm. In the second study we tested the effect of three doses on Results: Two hundred and sixty general practitioners (210 men and 50 women) digital skin blood flow (quantified using laser speckle contrast imaging). In the included 1379 patients with 28% men and 72% women. The details of analge- third study, the dose selected in study 2 is tested in twelve SSc patients. Blood sics’ prescriptions are presented in another poster. pressure was recorded continuously during all experiments and data were Male GPs prescribe more grade2 analgesics and more drugs against neuropathic expressed as cutaneous vascular conductance (CVC). pain than female GPs (0.35 vs. 0.27 grade2 analgesics per patient, P = 0.03 and Results: Iontophoresis of treprostinil increases CVC on the forearm (P = 0.02 vs. 0.13 vs. 0.06 drugs against neuropathic pain per patient, P = 0.02). Female GPs NaCl). Maximal intradermal concentration of treprostinil were obtained at H2 prescribe more antiarthrosic drugs (0.23 vs. 0.12 per patient, P < 0.0001). and were below the detection threshold in all but two participants at H8, while However, patients of male GPs have more intense pain than those of female GPs plasma concentrations were <1.7 pg/mL at all points for all subjects. Among the using auto and hetero-assessment scales. Patients of male GPs are significantly three doses tested on the finger pad, iontophoresis at 240 mC/cm² induced a sig-

nificant increase in CVC (P = 0.01 vs. NaCl). There was no local or systemic 07-01 adverse event. Digital iontophoresis at 240 mC/cm² is being tested in SSc Interest of adding Ab(1–40) determination in non-univocal profile of patients. cerebrospinal fluid biomarkers of Alzheimer disease Conclusion: Iontophoresis allows non-invasive administration of treprostinil into F Lemaitrea, P Lelievrea, B Langreea, S Belliardb, E Bellissanta, the dermis, without systemic diffusion of the drug. Digital cathodal iontophoresis MC Verdiera aLaboratory of Experimental and Clinical Pharmacology, Department of at 240 mC/cm² significantly increases skin blood flow on the finger pad in Clinical and Biological Pharmacology & Pharmacovigilance, Faculty of Medicine, healthy volunteers. Iontophoresis of treprostinil may be a way of avoiding sys- Pharmacoepidemiology and Drug Information Center, Rennes University Hospital, temic side effects of prostacyclin analogues in SSc patients. Rennes 1 University, Rennes, France; bDepartment of Neurology, Rennes University Reference: Hospital, Rennes, France 1. Blaise S., et al. J. Clin. Pharmacol. (2012). In press. Introduction: Alzheimer disease (AD) diagnosis is based on clinical signs, neu- Keywords: Iontophoresis, treprostinil, systemic sclerosis, digital ulcers, microdial- roimaging and cerebrospinal fluid (CSF) biomarkers. AD patients generally display ysis. decreased CSF Ab(1–42) (<500 pg/mL) and increased total Tau protein (H-Tau, >450 pg/mL) and phosphorylated Tau protein (p-Tau, >60 pg/mL). Decision algorithms using the combination of an index (Innotest Amyloid Tau Index, IATI = Ab(1–42)/(240 + 1.18 9 H-Tau) and p-Tau level exist but some CSF pro- SESSION 7: GT NEUROPSYCHO-PHARMACO files remain non-univocal due to an unexpected level of Ab(1–42). Ab(1–40) is an additional biomarker of beta-amyloid (bA) production which could define the relative abundance of Ab(1–42) according to the whole bA production. The aim Serotonergic receptor ligands: an open avenue for the treatment of of this study was to evaluate its interest to make a decision in patients with non- memory disorders? a a univocal profiles of CSF biomarkers of AD. F Dauphin Normandie Univ, Groupe Memoire et Plasticite comportementale, Caen, Methods: Sixty-six patients were retrospectively studied. Twenty-five patients France had normal CSF Ab(1–42) and elevated p-Tau and H-Tau. These patients had an Though cholinesterase inhibitors and memantine are still proposed for the symp- AD CSF profile with normal Ab(1–42) (Group I). Forty-one patients had tomatic treatment of memory deficits in Alzheimer’s disease, there is a crucial decreased Ab(1–42) and normal p-Tau and H-tau. These patients had a non-AD need to set up new and really efficient therapeutic strategies that could counter- CSF profile with decreased Ab(1–42) (Group II). The 66 patients had an Ab(1– balance at least the early alterations of episodic and working memory that are 40) determination and the Ab(1–42)/Ab(1–40) ratio was calculated. A ratio associated to normal and pathological aging as well as to psychoses. With this in <0.05 was considered in favor of AD. mind, serotonin receptors (5-HTR) represent promising therapeutic targets since Results: Among patients of group I, 11 (44%) had a ratio <0.05. These patients the serotoninergic neurotransmission system is implicated in the modulation of were considered as naturally high producers of bA and their Ab(1–42) seemed learning and memory processes. Among 5-HT receptors, 5-HT4, 5-HT6 and 5- falsely normal. In group II, 34 (76%) had a ratio above 0.05. The large majority HT7 receptors (5-HT4R, 5-HT6R and 5-HT7R) have received specific attention in of these patients were naturally low producers of Ab(1–42) and their Ab(1–42) the last decade, mainly on the basis of their central distribution in brain regions seemed falsely decreased. that are known to be implicated in learning and memory. Pre- or post-acquisition Discussion – Conclusion: While the determination of Ab(1–40) seemed to con- blockade of 5-HT6R in young adult mice prolonged the duration of episodic-like firm the diagnosis in group II patients, it added confusion in interpreting the memory (place recognition test in the Y maze) when compared to saline-treated results of group I patients. Ab(1–40) added few in the decision algorithm in animals. Furthermore, aged mice, which expressed spatial memory deficits, patients with normal Ab(1–42) levels and pathological p-Tau and H-Tau levels. explored the new arm more after a 60-min ITI (21-month-old) and a 240-min On the other hand, it helped to confirm the non-pathological low production of ITI (18- and 21-month-old) when treated with the 5-HT6R antagonist SB- Ab(1–42) in a majority of patients with non-definitive biological diagnosis. These 271046. Consequently, 5-HT6R blockade improves spatial recognition memory results have to be confirmed with clinical and radiological assessments. in adult mice and reverses age-related consolidation deficits of episodic-like mem- Keywords: Alzheimer disease, beta-amyloid, Cerebrospinal fluid, CSF, Biomar- ory. Deficits in consolidation of nonspatial recognition memory (object recogni- kers, Abeta 42, Abeta 40. tion test) that were observed in 17- and 21-month-old mice were also found to be reversed by 5-HT6R blockade. Deficits in working memory performances were only apparent as late as at 25 months of age; again, these deficits were reversed 07-02 by 5-HT6R blockade. This study revealed in the mouse that, as in humans, work- Higher insulin levels at baseline are associated with greater huntington’s ing memory is more lately altered than recognition memory during aging and disease deterioration that such memory deficits could be counteracted by the use of 5-HT6R antago- N Saleha, L Salema, G Desamericqa, K Youssova, G Dolbeaua, L Cleret De nists. We have further shown that the modulation between 5-HT6R and the cho- Langavanta, ML Bourhisb, S Moutereaub, AC Bachoud-Levia, P Maisona aInserm linergic system appears to be predominant for working memory and aversive U955 equipe 01 ‘Neuropsychologie Interventionnelle’, Creteil, France; bAP-HP, Groupe learning, but not for episodic-like memory. In this latter case, interactions Hospitalier Chenevier-Mondor, Creteil, France between 5-HT6R and alternative neurotransmission systems (i.e. glutamatergic Background and purpose: Recent studies have shown that the Insulin-like system) should be privileged. The respective involvement of these interactions, growth factor 1 (IGF-1) is related to neurodegenerative diseases such as Hunting- and the precise mechanisms of action (modulation of cAMP concentrations, … ton’s disease (HD) and may directly influence their progression. Given the struc- mTOR signaling, ), in the memory disorders related to ageing, neurodegenera- tural homology between IGF-1 and Insulin at both levels, molecules and tive diseases and psychiatric situations are of pivotal importance regarding the receptors, and since metabolic alterations related to Insulin were reported in both possible use of 5-HT6R antagonists in the treatment of memory disorders in patients and animal models of HD; we aim to examine the association between humans. Insulin plasma level and severity of HD progression independently to IGF-1. Methods: Eighty-four patients with genetically documented HD, at all stages of the illness, aged between 22 and 77 years, were studied for a follow-up period of approximately 3 years. We determined fasting blood levels of total Insulin at baseline. The Unified Huntington Disease Rating Scale (UHDRS) was used to Multiple system atrophy: the best clinical model to study disease- assess clinical impairment at both baseline and follow-up period. Longitudinal modifying therapeutic strategies in synucleinopathies? relationships were assessed using mixed-effects linear model. W Meissnera aUMR 5293 – Service de Neurologie, Centre Expert Parkinson et Centre – Results: In a multivariate cross-sectional analysis, adjusted for age, sex, neuro- de reference AMS, Institut des Maladies Neurodegeneratives, CHU de Bordeaux leptic treatment, age at onset, body mass index, CAG repeat and level of educa- CNRS, UMR 5293, Univ. de Bordeaux, Bordeaux, France tion for cognitive scores, higher plasma Insulin concentrations at baseline were Multiple system atrophy (MSA) is a rapidly progressing atypical parkinsonian dis- significantly associated with greater impairment on all UHDRS scores except for order leading to severe motor disability and death after only 9 years from symp- the Total Motor Score and the Symbol Digit Modalities test. In longitudinal analy- tom-onset. Together with Parkinson’s disease (PD) where clinical signs are slowly sis, all UHDRS scores, except for behavioural score, declined significantly over progressing over many years, MSA belongs to a group of neurodegenerative dis- time; but no significant association was found between baseline insulin levels and orders, the alpha-synucleionopathies, which are characterized by the abnormal UHDRS score deterioration over time. accumulation of alpha-synuclein. In contrast to PD, alpha-synuclein mainly Conclusion: Our findings show that higher Insulin level at baseline is associated accumulates in glial cells but also in neurons. In recent years, crucial milestones with greater impairment in HD. However, despite structural homology between have been reached for successfully conducting clinical intervention trials in MSA Insulin and IGF-1, Insulin does not predict the disease progression unlike IGF-1 patients. For example, consensus diagnosis criteria are now available, clinical which predicts cognitive evolution. scales assessing different aspects of motor impairment, activities of daily living Keywords: Huntington’s Disease, Insulin, Insulin-like growth factor 1. and quality of life have been validated, and large international networks have been established in Europe, North America and Asia. Moreover, clinical intervention trials were able to enrol as much as 400 MSA patients and animal models of MSA that recapitulate hallmark features of human disease enable rapid screen- 07-03 ing for potential drugs and characterization of underlying disease mechanisms. Drugs related anticholinergic burden and delirium in elderly patients M Najaa, P Jouannya, S Hannata, J Zmudkaa, M Andrejakb, S Liabeufb aCHU Because there are no objective biomarkers available, endpoints of disease modify- b ing/neuroprotective trials in MSA and PD are relying on clinical rating scales. AMIENS Service de medecine geriatrique, Amiens, France; CHU Amiens Service de Since disease progression is more rapid in MSA than in PD and potential con- pharmacologie clinique – Centre regional de pharmacovigilance, Amiens, France founding effects of symptomatic treatments are rarer, MSA could prove an advan- Objective: Polymedication is frequent in elderly and lead to a high prevalence of tageous model to assess disease modifying/neuroprotective strategies in alpha- iatrogenic risk. Indeed, anticholinergic agents can induce potential central ner- synucleinopathies. vous system adverse reactions. So the aim of this study was to evaluate the use of anticholinergic drugs and its impact on delirium. Patients and methods: Ninety-one consecutive patients hospitalized in geriatric department of Amiens university hospital were evaluated. Prescriptions at home and during hospitalization were analyzed and anticholinergic burden was classified into four levels (none, low, medium and high) every week. Delirium symptoms were measured with Confusion Assessment Method (CAM) at day 1, 3, 5, 8, 15 and 21. Covariates studied are polypathology (Charlson score), health status, autonomy (ADL / IADL), nutritional status (albumin), cognition (MMS),

length of stay and mortality. Univariate and multivariate analyses have been per- Results: Five hundred and ninety-four subjects were included; 63.6% living at formed to identify factors associated with delirium symptoms. home and 36.4% in institution. Mean age was 84.2 Æ 5.0 years (75–99) and Results: Ninety-one patients (86.3 Æ 5.8 years, 53 women and 49 men) were 73.7% of subjects were women. Mean MMSE was 16.9 Æ 6.0 (0–29); Alzhei- included and followed up for 14.5 Æ 9.9 days. 51.2% were taking drugs with mer’s disease represented 74.9% of patients. The mean number of drugs anticholinergic properties at home. Anticholinergic burden average is (6.6 Æ 2.8) and the PIM use prevalence [44.6% (95%CI: 40.6–48.7%)] were not 2.13 Æ 1.34. Prevalence of delirium symptoms were respectively: 43.1 % at Day significantly different between the metropolitan (44.1%) and the over-seas areas 1, 36.9 % at Day 3, 34.8 % at Day 5, 44.9 % at Day 8 and 60 % at Day 21. (51.1%). The most often used PIM were (87.5%): benzodiazepines, association of Patients with delirium had higher mortality (16.1% vs. 3.7 %; P = 0.049), psychotropics, cerebral vasodilators, association of anticholinesterasics and anti- higher average length of stay (18.09 Æ 11.34 vs. 11.75 Æ 7.80 days; cholinergics. The risk factors of PIM use were polymedication (OR = 4.0, 95%CI: P = 0.001), decreased autonomy on discharge (ADL 1.57 Æ 1.56 vs. 2.5–6.4), life in institution (OR = 1.9, 95%CI: 1.3–2.7) and being a woman 3.41 Æ 1.45 on 6; P < 0.0001) and had health status more degraded on dis- (OR = 1.5, 95%CI: 1.0–2.2). charge (5.3 Æ 2.7 vs. 7.0 Æ 1.7 on 10; P = 0.0008). Delirium symptoms were Conclusion: The use of PIM is high in the French population of patients with correlated with high anticholinergic burden (P = 0.017). Anticholinergic burden Alzheimer’s disease and related dementia. These drugs should be used cautiously was independently associated with delirium symptoms as evaluated by CAM. in these patients as they could aggravate memory and dementia. Discussion: We demonstrated in hospitalized elderly patients that anticholinergic Keywords: Alzheimer’s disease, inappropriate medication, misuse, France, multi- burden is a definite risk factor for delirium. Since delirium can lead to marked centre observational study. alterations in global health status and could be associated with mortality, the limitation of anticholinergic burden seems crucial in elderly people. Keywords: Anticholinergic burden, delirium, geriatric population. 07-06 Long-term efficacy of antipsychotics for neurocognitive deficits in schizophrenia: network meta-analysis 07-04 G Desamericqa, A Mearya, I Macquin-Maviera, AC Bachoud-Levia,P Psychotropic drugs use and misuse in employees of a regional Maisonb aAP-HP, Creteil Cedex, France; bINSERM, Creteil, France psychiatric hospital: prevalence and factors associated Objectives: We conducted a network meta-analysis on all the randomized con- K Lherminier-Miharaa, J Dupouyb, JC Poutrainc, S Oustricc, M Lapeyre-Mestred, trolled trials in order to classify antipsychotic drugs on cognitive outcomes in N Jobe aDepartement Universitaire de Medecine Generale, Centre Hospitalier Gerard patients with schizophrenia. Marchant, Toulouse, France; bEquipe de Pharmacoepidemiologie, Departement Material and methods: To identify relevant publications, we searched (without Universitaire de Medecine Generale, UMR 1027 Inserm-UPS, Toulouse, France; language restrictions) MEDLINE and EMBASE (up to week 3 August 2011) for cDepartement Universitaire de Medecine Generale, Toulouse, France; dEquipe de randomized controlled trials (RCTs) in which oral formulations of second-genera- Pharmacoepidemiologie, UMR 1027 Inserm-UPS, Toulouse, France; eCentre tion antipsychotic drugs (amisulpride, aripiprazole, olanzapine, quetiapine, risperi- Hospitalier Gerard Marchant, Toulouse, France done, sertindole, ziprasidone, and zotepine) were compared to placebo or Background: Psychotropic drugs are known to be widely used in workers. Few haloperidol or second-generation antipsychotic drugs, for the treatment of schizo- studies have assessed psychotropic drugs misuse in hospitals employees in France. phrenia or related disorders. Trials were required to be of at least 6 months dura- Availability of psychotropic drugs, working by night, job strain, personal history tion and were combined using mixed treatment comparisons models to provide are known risk factors for psychotropic drugs misuse. Psychotropic drugs misuse direct and indirect comparisons in a single analysis. in employees of psychiatric hospitals might be substantial. Results: Nine studies were eligible. Quetiapine, olanzapine and risperidone had Aim: We conducted a transversal descriptive study in a regional psychiatric hos- better effect on global cognitive score than amisulpride (mean difference MD pital (610 beds), aiming to describe prevalence of psychotropic drugs misuse. Our 0.27, 95% CI 0.10–0.44; MD 0.20, 95% CI 0.04–0.37; MD 0.16, 95% CI À0.03 secondary aim was to assess factors associated with psychotropic drugs misuse. to 0.34, respectively) and haloperidol (MD 0.27, 95% CI 0.13–0.41; MD 0.21, Method: A self-administrated anonymous questionnaire was send by internal 95% CI 0.10–0.32; MD 0.16, 95% CI 0.02–0.30). When memory was consid- mail in May 2011 to all employees of the psychiatric hospital in the Toulouse ered, ziprasidone had better effect than amisulpride (MD 0.28, 95% CI 0.02– city. No exclusion criterion was retained. Data collected were socio-demographic 0.54) and haloperidol (MD 0.32, 95% CI 0.09–0.55). On attention and process- data, medical data, use of psychotropic drugs (prescribed by a physician) or mis- ing speed, quetiapine was largely better than others drugs (P < 0.001), followed use (defined by using psychotropic drugs without prescription), drug use, per- by ziprasidone (P < 0.05) and olanzapine (P < 0.05). Quetiapine, risperidone and sonal and familial previous history of substance abuse. Severity of substance olanzapine were the best antipsychotics (P < 0.05 vs. amisulpride) on executive abuse was assessed by the Drug Abuse Screening Test (DAST) 10. Job strain was function. Theses finding were not modified when only patients in their first epi- assessed by the French version of the Karasek Job Content Questionnaire. A logis- sode were considered tic regression was conducted to assess factors associated with this misuse. Discussion: Significant differences exist between medications on global with Results: Of the 1326 questionnaires send, 459 (34.6%) were returned and ana- highest effect for quetiapine and olanzapine, followed by risperidone and ziprasi- lyzed. Responders were mainly women (335, 73.8%) of median age 42 years done and finally amisulpride and haloperidol. Significant differences were also [33; 52]. The most represented job was nurses (183, 39.9%); 145 (31.6%) observed according to the cognitive task. employees regularly worked by night. Thirty-seven responders (8.1%) declared to Keywords: meta-analysis; antipsychotic drugs; schizophrenia; cognition. be drug users. Fifty-six responders (12.2%) declared to use psychotropic drugs without prescription; these drugs were mainly hypnotics and anxiolytics, zolpi- dem being the most cited drug. According to DAST-10, 24 psychotropic drugs misusers (42.9%) were screened as having ‘low’ or ‘moderate’ or ‘substantial CALCIFICATION DES TISSUS CONJONCTIFS: DE LA problem’. Fifteen psychotropic drugs misusers (26.8%) were suffering from job strain. In multivariate analysis, factors associated with psychotropic drug misuse GEN ETIQUE A LA PATHOLOGIE were to be a woman, to work by night, to be treated with psychotropic drugs prescribed by a physician and to have a personal history of drug use. c Discussion: Psychotropic drug misuse was substantial in employees of a psychi- -glutamyl carboxylase regulates whole body energy metabolism through atric hospital. Information and screening must play a key role for prevention. its expression in osteoblasts M Ferrona, G Karsentyb aUnite de recherche en physiologie integrative et moleculaire, Keywords: Psychotropic Drugs, Psychiatric Hospital, Health Personnel, Sub- stance-Related Disorders. Department of Genetics and Development, Institut de Recherches Cliniques de Montreal, Columbia University, 701 W 168th Street, New York, NY, USA; bDepartment of Genetics and Development, College of Physicians and Surgeons, Columbia University, New York, NY, USA 07-05 Osteocalcin is a hormone produced by osteoblasts that regulates energy metabo- Potentially inappropriate medications in the elderly with Alzheimer’s lism by enhancing insulin secretion and insulin sensitivity. While osteocalcin can disease and related dementia in France: results of the MIDA study exist in two forms, c-carboxylated and undercarboxylated, only the undercarb- M Lassallea, JP Charmesb, J Doucetc, M Rainfrayd, R Gonthiere, A Lahlouf, g h a a oxylated form of this molecule appears to function as a hormone. Thus, the E Alix , JM Eychenne , ML Laroche Centre of Pharmacovigilance, EA HAVAE, results obtained so far imply that c-carboxylation of osteocalcin should have an University Hospital, Limoges, France; bALURAD, Limoges, France; cGeriatrics d inhibitory effect on the bioavailability of this hormone and, indirectly, on whole Department, University Hospital of Rouen, Saint-Julien, France; Geriatrics body glucose homeostasis. To determine if it is the case in vivo we generated Department, University Hospital of Bordeaux, Pessac, France; eGeriatrics Department, c À/À f osteoblast-specific -glutamyl carboxylase-deficient mice (Ggcxosb ). In contrast University Hospital of Saint-Etienne, Saint-Etienne, France; Geriatrics Department, to the Ggcx null mice, which died shortly after birth form haemorrhage, the AP-HP, Ivry-sur-Seine, France; gGeriatrics Unit, Hospital, Le Mans, France; À/À h Ggcxosb mice were viable and obtained at the expected Mendelian ratio. These Geriatrics Department, Hospital, Saint-Joseph, La Reunion, France mutant mice however, displayed a fivefold increase in their serum levels of un- Objective: Alzheimer’s disease and other dementia are mainly encountered in carboxylated osteocalcin. This resulted in a significant improvement of glucose the elderly. These patients use numerous medications. Some of these drugs may À/À handling in Ggcxosb mice as measured by a glucose tolerance test (GTT). An be considered as potentially inappropriate if their benefit/risk ratio is not judged insulin tolerance test (ITT) revealed that insulin sensitivity was also improved in favourable in this frail and old population. A list of potentially inappropriate med- À/À these mutant mice. Lastly, Ggcxosb mice presented a 50% decrease in their fat À/À ications (PIM) has been developed inFrance on the basis of an expert consensus. mass. Importantly, when fed a high fat diet the Ggcxosb mice gained signifi- This list is considered as an epidemiological tool to assess the quality of prescrip- cantly less weight than control mice and were protected against glucose intoler- tion in geriatrics or in any old sub-population. The aim of the study was to assess ance. Taken together, these results establish that c-glutamyl carboxylase the prevalence and the risk factors of PIM use in the elderly with Alzheimer’s dis- negatively regulates glucose homeostasis through it expression in osteoblasts. ease and related dementia in France. This work further underscores the importance of bone in glucose homeostasis Methods: A French cross-sectional observational multicentre study was con- and suggests that Vitamin K, an essential cofactor of c-glutamyl carboxylase, ducted from 1/2009 to 6/2011 in six metropolitan regions (Aquitaine, Haute- ^ may play a role in the control of energy metabolism by bone. Normandie, Ile-de-France, Limousin, Pays de la Loire, Rhone-Alpes)^ and one Keywords: c-Glutamyl. over-sea region (La Reunion). Patients aged 75 years and more with dementia, treated with cholinesterase inhibitors or memantine were included. They were selected in medical structures according to their usual dwelling place (at home or institution). Socio-demographic data, history, drugs given were registered. PIM use was estimated using the French PIM list. (Funding: PHRC national 2008).

INFLAMMATION ET PATHOLOGIES BRONCHIQUES ing interventions were widely proposed as an antiaging strategy in humans. For instance, antioxidants, a group of synthetic or naturally occurring substances, Bronchial inflammation in asthma: interaction mast cell – airway which are capable of scavenging ROS, are largely studied and used as agents to smooth muscle cell delay aging. Unexpectedly and in contrast to some works, epidemiological studies R MARTHANa aInserm U1045, Centre de Recherche Cardio-Thoracique, Universitede were unable to show a positive association between supplementation with antiox- Bordeaux, Bordeaux, France idants and health-beneficial effects. Indeed, most studies found a lack of effect in There is strong evidence that, in asthma, mast cells infiltrate the airway mucosa regards to health promotion in humans, whereas other reports even suggest that and, as a consequence, alter both functional and structural properties of airway antioxidants may promote tumor development. Finally, supplementation with an- smooth muscle cells. On the one hand, the major product of the mast cell, the tioxidants has been linked to increased incidence of a number of diseases with trypsin-like serine proteinase, tryptase, has been shown to induce airway hyper- adverse effects on human longevity. Here, we will show how recent evidences responsiveness and airway smooth muscle cell proliferation via a calcium depen- indicate that, one the one hand, ROS may evoke cellular signaling that promotes dent pathway (1, 2). On the other hand, human and experimental data indicate metabolic health and longevity, and on the other hand, low levels of ROS may that, upon stimulation by mast cell-derived tryptase, smooth muscle cells can prevent or limit inflammation and could possibly have a role in resolution of attract mast cells (1). In this connection, it has been demonstrated that the num- inflammation. ber of mast cells present in the smooth muscle layer of the bronchial wall is significantly higher in asthmatic patients than in normal subjects. Moreover, a variety of chemokines produced by stimulated airway smooth cells from asthmatic patients has been shown to induce mast cell chemotaxis thus generating an auto-activation loop that favors mast cell survival and degranulation. Mast Oxidative stress and Redox regulation: The dual role of mitochondrial cells infiltrating the smooth muscle layer are not in direct contact with airway Ros G Simarda aLaboratoire de Biochimie, INSERM UMR1063, Institut de Biologie en smooth muscle cells but in a very close relationship with the extracellular matrix (3) and recent data obtained using blocking antibodies suggest that CD44 and Sante, 4 rue Larrey, Angers, France CD51 are responsible for mast cell adhesion to airway smooth muscle (4). Mitochondria synthesize ATP through a regulated process of oxidative phosphor- Most of these effects are mediated by Proteinase-Activated Receptor 2, (PAR-2) a ylation driven by electron transport across the electron transport chain (ETC). As member of a family of hepta-helical receptors activated upon the proteolytic a natural byproduct of metabolism, reactive oxygen species (ROS) are also generated when a small percentage of electrons leak out of the ETC, combine with O2 activity of enzymes and expressed by airway smooth muscle cells. Alternative . compounds such as the chitinase 3-like 1 protein, YKL-40, that has been evoked and form superoxide anions (O2 -). The two major sites of superoxide production in asthma, also increase airway smooth muscle cell proliferation through PAR-2- are at complex I and III and under physiological conditions, it is estimated that dependent mechanisms (5). 0.2% of the oxygen consumed by the cells is converted into the superoxide anion. This auto-activation loop implicating mast cell-derived products that activate air- The mitochondrial ROS production is rapidly neutralized by efficient antioxidant systems, including superoxide dismutases (SOD) that catalyze the dismutation of way smooth muscle cells that, in turn, attract mast cells to airway smooth mus- . cle cells may provide several relevant therapeutic targets for the treatment of O2 - to hydrogen peroxide H2O2 and the catalase and glutathione peroxydases asthma, some of them being related to modulation of calcium homeostasis in air- that reduce and inactivate H2O2 to water. way smooth muscle (6). When ROS generation overwhelms the antioxidant defense, the radicals can References: interact with endogenous molecules and alter cellular function. Excess mitochon- – drial ROS production may occur when mitochondria is directly affected or 1. Berger P., Girodet P.O., Begueret H., et al. Faseb. J. (2003) 17 2139 2141. … 2. Trian T., Benard G., Begueret H., et al. J. Exp. Med. (2007) 204 3173–3181. involved in a disease state (aging, diabetes mellitus, ETC defect, obesity .) but 3. Begueret H., Berger P., Vernejoux J.M., et al. Thorax (2007) 62 8–15. also when cytosolic ROS concentration is increased. In recent years, a number of 4. Girodet P.O., Ozier A., Trian, et al. Allergy (2010) 65 1004–1012. studies have suggested that mitochondria are an important constituent of a posi- 5. Bara I., Ozier A., Girodet P.O., et al. Am. J. Res. Crit. Care Med. (2012) 185 tive feedback leading to an amplification of ROS production (concept of ROS-tri- 715–722. gerred ROS). A good example is the role of NADPH oxidase-derived cytosolic ROS 6. Girodet P.O., Ozier A., Carvalho G., et al. Am. J. Res. Cell Mol. Biol. (2013) 48 in triggering mitochondrial ROS formation in vascular dysfunction. 212–219. On the other hand, several lines of research have demonstrated the potential ben- Keywords: muscle cells. eficial role of mitochondrial ROS. As the major source of ROS in cell, mitochondria contribute to redox signalling. Hydrogen peroxide released by mitochondria may modulate the activity of targets proteins by oxidizing redox sensitive thiols and is an important regulator of eukaryotic signal transduction. Mitochondria redox signalling participates also in oxygen sensing and the hypoxia induced STRESS OXYDATIF: LES ESPECES RADICALAIRES release ROS leads to an increased expression of hypoxia sensitive genes such as VEGF which in turn mediate adaptive metabolic responses. Mitochondrial ROS are SONT-ELLES DE NOUVELLES CIBLES also involved in cardioprotective signalling in the context of ischemia/reperfusion. PHARMACOLOGIQUES? Hence mitochondrial ROS production can have both negative (damage) and positive effects (redox signalling). This should be taken into consideration in treatment strategy. Treatments that raise the levels of antioxidants are also likely to Do free radical species represent new pharmacological targets? The pros a a disrupt important redox signalling. Another interesting strategy is to target mito- L Liaudet Department of Intensive Care Medicine and Burn Center, Faculty of chondria to limit its ‘bad’ ROS production. According to this view, the control of Biology and Medicine, University Hospital Medical Center, Lausanne, Switzerland mitochondrial channels such as mPTP or the mtK are promising strategies. Free radical chemical species, either oxygen-centered (reactive oxygen species) or ATP nitrogen-centered (reactive nitrogen species), are produced physiologically by multiple enzymatic and non enzymatic systems and play an important role in cellular homeostasis by regulating numerous signal transduction pathways (redox signaling). These processes are guaranteed through the maintenance of redox SESSION 11: GT CARDIOVASCULAIRE equilibrium, ensured by an appropriate balance between radical species and redox buffers (antioxidants). In multiple pathological conditions, this balance is PCSK9: a new target for hypercholesterolemia interrupted following the generation of ROS/RNS in excess of the physiological B Carioua aClinique d’Endocrinologie-INSERM UMR1087, l’Institut du Thorax, antioxidant buffering capacities, inducing a state of redox stress, the nature of Nantes, France which may be oxidative, nitrosative or nitroxidative. In such conditions, radical PCSK9 (Proprotein Convertase Subtilisin Kexin type 9) is a secreted protease that species produced in large amounts may exert significant deleterious effects by modulates cholesterol homeostasis by decreasing LDL receptor (LDLr) expression. inflicting various damages to major biomolecules (lipids, protein and nucleic PCSK9 gain-of-function mutations within PCSK9 are linked to familial autosomal acids), leading to cellular dysfunction, irreversible cellular injury (necrosis, apop- dominant hypercholesterolemia, characterized by elevated plasma concentrations tosis) and fostering the development and the amplification of inflammatory proof cholesterol associated to low density lipoproteins (LDL-C). Conversely, loss-of- cesses. It has been therefore clearly established that redox stress plays a critical function mutations result in low LDL-C levels and protect against coronary heart role in the occurrence of tissue injury in both acute (for instance ischemia-reper- disease. Although compelling evidence suggests that PCSK9 impairs the LDLr fusion, circulatory shock, trauma) and chronic pathological conditions (such as pathway, its role on cholesterol metabolism remains incompletely defined. In the cardiovascular diseases, diabetes, neurodegeneration, chronic inflammatory dis- past 2 years, several new biochemical findings were reported, including PCSK9 eases, and cancer). In such conditions, reducing redox stress by therapies aiming crystal structure and the identification of several transcriptional repressors. More- either at the direct elimination of radical species or at the inhibition of the mech- over, new clinical and epidemiological data accumulated, in particular concern- anisms involved in their production, may prove therapeutically useful, a concept ing the correlation between plasma PCSK9 concentrations and LDL-C levels. supported by multiples lines of experimental evidence. We will review the main More importantly, new pharmacological strategies aiming for PCSK9 inhibition forms of chemical radical species, their mechanisms of formation, their chemical have arisen during the last year. Notably, phase 2 studies with monoclonal anti- biology and their pathological actions in some illustrative situations, in order to bodies directed against PCSK9 have led to consistent and promising results in demonstrate the pharmacological interest to suppress redox stress in such condi- patients with either heterozygous familial hypercholesterolemia or primary hyper- tions. cholesterolemia with a 50–60% reduction of LDL-C levels in association to statin therapy. Phase 3 studies are currently ongoing and will help to position PCSK9 inhibitors in the pharmacological management of hypercholesterolemia. Do free radical species represent new pharmacological targets? The cons C Martineza aINSERM UMR 1063, Stress Oxydant et Pathologies Metaboliques, Institut de Biologie en Sante, Angers, France Although reactive oxygen species (ROS) have not traditionally been studied as ‘signaling’ molecules, alteration in cellular redox status, by slight imbalances in CETP inhibition: a strategy to abandon? ROS generation and elimination, is now recognized as an important mediator of A Kontusha aINSERM Research Unit 939, Hopital de la Pitie, University of Pierre various cellular functions. An exacerbated increased in ROS formation, as a con- and Marie Curie – Paris 6, Paris sequence of increased metabolic rate, has been proposed to be the key determi- Plasma high-density lipoprotein (HDL) is a small, dense and protein-rich lipopro- nant of life span suggesting that reduction of oxidative stress is associated with tein as compared to other lipoprotein classes. HDL metabolism is thought to prolongation of life expectancy in various organisms. Consequently, ROS-lower- underlie the processes of cellular cholesterol efflux and, in part, reverse choles-

terol transport (RCT) from the arterial wall to the liver for excretion into the bile. current studies aim at identifying the mechanisms involved in this beneficial met- Consistent with the concept of RCT, low circulating levels of HDL-cholesterol abolic action. (HDL-C) are an independent predictor of cardiovascular disease. Methods: Fourteen months-old male Wistar rats were treated for 1 week with Cholesteryl ester transfer protein (CETP) is a 74 kDa protein which facilitates the LNP509 (10 mg/kg/day ip) or vehicle. Insulin sensitivity was then evaluated in bidirectional transfer of cholesteryl esters and triglycerides between HDL and pro- anesthetized animals by measuring the hypoglycemic action of insulin (1.5 U/kg atherogenic apolipoprotein (apo) B-containing lipoproteins. Elevated CETP activity ip). Since adiponectin is a major insulin-sensitizing hormone, LNP509-induced thereby results in reduced levels of HDL-C; hence, the concept of CETP inhibition changes in the plasma level of the adipokine were sought; density of adiponectin to reduce cardiovascular risk. receptors in hepatic tissue was also assessed. CETP inhibitors are small molecules which inhibit its capacity to facilitate hetero- Results: Insulin injection produced marked and long lasting hypoglycemia in all transfer of neutral lipids between HDL and apoB-containing lipoproteins. Three animals, but the effect was more pronounced in LNP509-treated rats. Thus, small-molecule inhibitors of CETP, torcetrapib, dalcetrapib (JTT-705) and anacet- plasma glucose was reduced by 40–45% after 30 min and remained stable there- rapib (MK-0859), have initially been developed which induce elevation in con- after in vehicle-treated rats (6.6 Æ 0.4 and 3.8 Æ 0.2 mM at t0 and t30 min, centrations of HDL-C while decreasing those of low-density lipoprotein-cholesterol respectively); in contrast, glycemia continued to decrease in rats receiving (LDL-C) and apoB. CETP inhibitors are presently the most potent HDL-raising LNP509, to reach a 60% reduction after 60 min (6.6 Æ 0.1, 3.2 Æ 0.1 and agents, with dose-dependent HDL-C elevation of up to +100% or more for some 2.6 Æ 0.2 mM at t0,t30 min and t60 min, respectively). Consequently, the area agents. A CETP vaccine which induces autoantibodies that specifically bind to under the curve of glycemia over time was lower in LNP509-treated rats endogenous CETP represents another approach to inhibit CETP activity. (214 Æ 7 vs. 247 Æ 14; P < 0.05). LNP509 significantly increased the plasma Torcetrapib became the first CETP inhibitor to enter clinical trials and revealed adiponectin concentration (2.04 Æ 0.21 mg/L in treated rats vs. its marked efficacy in improving an atherogenic plasma lipid profile. The first 1.48 Æ 0.11 mg/L in control rats) as well as adiponectin receptor densities in large-scale, prospective, placebo-controlled interventional ILLUMINATE trial hepatic tissue (adipoR1: 40.4 Æ 5.2 and 27.8 Æ 2.5 ng/mg protein in treated assessing the impact of a CETP inhibitor on cardiovascular risk was however pre- and control rats, respectively; adipoR2: 1.9 Æ 0.2 and 1.2 Æ 0.2 ng/mg protein maturely terminated in December 2006 as a consequenceof excess mortality in in treated and control rats, respectively). the treatment group relative to placebo. More recently, clinical development of Conclusion: These data show that an imidazoline I1 receptor selective ligand dalcetrapib was stopped as a result of futility. These findings challenge the con- ameliorates insulin sensitivity of target tissues, thus improving glucose tolerance. cept of CETP inhibition to lower cardiovascular risk. How this is causally related to the concomitant stimulation of adiponectin path- Torcetrapib treatment was however associated with significant increment in aldo- way has to be further investigated. The mechanisms involved in the imidazoline- sterone levels, with changes in serum electrolyte levels indicative of mineralocor- like drugs-induced activation of adiponectin signalling are under investigation. ticoid excess, and with elevation of blood pressure. The aldosterone-mediated Keywords: adiponectin, imidazoline-like drugs, insulino-resistance, metabolic pressor effect of torcetrapib appears to be unrelated to HDL-C raising as indicated syndrome, sympatho-inhibition. by data obtained with other CETP inhibitors. These findings suggest an off-target mechanism of torcetrapib toxicity involving activation of mineralocorticoid receptors by aldosterone with subsequent induction of hypertension. 11-03 Despite the failures of torcetrapib and dalcetrapib, CETP inhibitors still appear to Evidence suggests that extracellular NAADP signaling at the P2Y11-like hold promise in terms of their ability to provide cardiovascular risk reduction. receptor affords cardioprotection through cyclic AMP and IP3 The net action of CETP inhibitors on RCT is however neutral in CETP-expressing Z Djeradaa, H Peyreta, H Millarta aDepartment of Pharmacology, E.A.3801, SFR species. Whereas the capacity of HDL to efflux and transport cholesterol does not CAP-sante, Reims University Hospital, Reims, France appear to be enhanced by CETP inhibitors, little is known of other atheroprotec- Objectives: Extracellular nucleotides may play important regulatory roles within tive activities of HDL. Ongoing clinical studies of novel CETP inhibitors will pro- the cardiovascular system and notably in cardioprotection. Firstly, we aimed to vide ultimate answers regarding the utility of this approach. look for possible cardioprotective effects following extracellular pre-treatment with [NAADP]e, an active metabolite of b-NAD+. Secondly, we characterized the involvement of a P2Y11-like receptor in mediating ischemic-reperfusion toler- 11-01 ance. Finally, we studied the effects of [NAADP]e on signal transduction mediated PTP1B genetic deletion limits myocardial dysfunction during endotoxinic by P2Y11-like and looked for complementary mechanisms of NAADP action. shock Material and methods: We tested whether pre-ischemic treatment with NAADP D Coquerela, X Marechalb, D Montaigneb, S Reneta, P Muldera, R Neviereb, associated or not with NF157, a P2Y11 antagonist, could enhance post-ischemic V Richarda, F Tamiona a INSERM 1096, Rouen, France; bEA 4484, Lille, France functional recovery, myocardial-necrosis and could decrease arrhythmogenesis, Sepsis-associated myocardial dysfunction is one of the main causes of mortality in an ex-vivo model of catecholamine-depleted rat hearts. Mechanistic studies in critically ill patients. PTP1B was describing as a negative regulator of insulin were performed on an in-vitro model of cardiomyocites. Signal transduction via signaling and endothelial Nitric Oxide (NO) production. We recently showed that Gq subunit of P2Y11-like receptor was evaluated with IP3 production. Signal PTP1B gene deletion or pharmacological inhibition afford myocardial protection transduction via Gs subunit of P2Y11-like was assessed with cAMP production in experimental heart failure. However, whether PTP1B inactivation opposes using chromatographic separation coupled with mass spectrometry detection LPS-induced myocardial dysfunction is unknown. (uPLC-MSMS). In the same time, other calcium intracellular mediators such as Thus, we assessed the effect of PTP1B genetic deletion on left ventricular (LV) cADP-ribose and intracellular [NAADP]i production were measured with uPLC- function by echocardiography and pressure-volume curves as well as ex-vivo in MSMS. Activation of the reperfusion injury salvage kinase (RISK) pathway (Akt, Langendorff isolated-perfused hearts. Inflammatory and oxidative stress markers ERK1/2 and GSK-3b) and PKCe with [NAADP]e was evaluated using western were assessed by Western blot and real-time RT-PCR finally mitochondrial func- blot. tion was measured by Respiratory Control Ratio (RCR). Results: Pre-treatment with [NAADP]e (0.1–1 lM) induced cardioprotective Compared to wild type (WT), PTP1BÀ/À mice showed a marked decrease in mor- effects with regards to functional recovery, necrosis and arrhythmogenesis tality (30% vs. 100%, n = 20) associated with a higher locomotor activity during (P < 0.05). These effects were completely suppressed with NF157. In cardiomyo- endotoxemia. In WT mice, LPS induced a severe LV dysfunction 8-h (H8) after cites cultures, [NAADP]e induced RISK pathways and PKCe activation LPS injection, as shown by the decreased fractional shortening (H0: 49 Æ 1%; (P < 0.01). These activations were suppressed by NF157, U73122 (phospholipase H8: 17 Æ 1%, P < 0.01) in association with a significant PTP1B overexpression. C inhibitor) and H89 (PKA inhibitor). [NAADP]e increased IP3 and cAMP pro- À/À In PTP1B mice, we demonstrated a significant improvement of both systolic duction with EC50 = 9.8 lM and EC50 = 0.18 lM respectively. Interestingly [NA- and diastolic LV function as shown in vivo by echocardiographic parameters and ADP]e induced an increase of two other calcium intracellular mediators i.e. by the LV End Diastolic and End Systolic Pressure-Volume Relation improvement [cADP-ribose]i and [NAADP]i production with the same EC50 = 15.7 lM. (LVESPVR WT H8 16.1 Æ 1.62 vs. PTP1BÀ/À H8 19.96 Æ 1.95 mmHg/RVU, Conclusion: Evidence suggests that NAADP receptor signalling at the P2Y11- P < 0.05).) but also by the higher LV Developed Pressure (LVDP WT H8 25 Æ 5 like receptor affords significant cardioprotection against ischemia and reperfusion vs. PTP1BÀ/À H8 50 Æ 5 mmHg) observed ex-vivo in Langendorff. These results injury. This protection probably involves the RISK pathway and PKCe activation were associated with a significant decrease of myocardial inflammatory state as induced by PLC and PKA. Furthermore, these results do not exclude a comple- shown by the lower mRNA expression level of VCAM-1, ICAM-1, CD45, IL-1b, mentary role of intracellular [NAADP]i and [cADP-ribose]i in cardioprotection TNF-a and Gp91phox in PTP1BÀ/À mice. Moreover we found that PTP1B genetic induced by extracellular NAADP. deletion limited P38 MAPK and ERK1/2 phosphosphorylation, well known to Keywords: NAADP, P2Y11- like receptor, cardioprotection, RISK pathway, promote pro-inflammatory cytokine production and accordingly major myocar- cADP-ribose, cAMP, IP3. dial depression. Finally, PTP1BÀ/À mice did not exhibit mitochondrial improvement compared to WT mice but had a limited impairment of SERCA2a/ Phospholamban regulated calcium homeostasis. 11-04 In conclusion, we demonstrated for the first time that PTP1B genetic deletion Role of angiotensin-converting enzyme 2 in resistance artery function limits LPS-induced LV dysfunction. Therefore, PTP1B could be a potential phar- M Brieta, R Touyzb, S Gurleyc, K Burnsb aHEGP, APHP, CIC9201, Paris, France; macological target for cardiovascular treatment during sepsis. bDivision of Nephrology, Department of Medicine, Kidney Research Centre, Ottawa Keywords: PTP-1B, Sepsis, myocardial dysfunction, inflammation. Hospital Research Institute, University of Ottawa, Ottawa, ON, Canada; cDivision of Nephrology, Department of Medicine, Duke University, Durham, NC, USA Background: Angiotensin-converting enzyme 2 (ACE2) metabolizes Ang II to 11-02 Ang-(1-7), which may have a protective role at the level of the cardiovascular Effects of imidazoline-like drugs on insulin sensitivity and adiponectin system through its action on the Mas receptor. We hypothesized that, in vivo, pathway ACE2 exerts a protective effect on resistance artery function. N Niederhoffera, S Bouchouchaa, M Weissa, H Greneya, P Bousqueta aUniversitede Methods: Ten to twelve-week old male ACE2 gene knockout (ace2-/y) and wild Strasbourg, Strasbourg, France type (WT) mice were implanted with a dummy pump (control) or infused with a Introduction: Metabolic syndrome is defined as a cluster of cardiovascular and subpressor doses of angiotensin II (400 ng/kg/min, s.c.) for 7 days (n = 6–8). metabolic disorders leading to increased cardiovascular risk. Several investiga- Systolic blood pressure (SBP) was measured by telemetry. Endothelial function tions showed that sympatho-inhibitory imidazoline-like drugs can improve gluci- and vessel structure were assessed in 2nd order mesenteric arteries by pressurized dic and lipid metabolism. In addition to their central effects, such drugs may also myography. Endothelial function was studied as the relaxation response to display peripheral direct metabolic effects. We demonstrated that a selective imi- increasing concentrations of acetylcholine (10–9to10–4 M) in the presence or dazoline I1 receptor ligand, LNP509, which does not cross the BBB, improves absence of Ang-(1-7) (10–7 M). glucose tolerance in aged rats displaying impaired glucose regulation. Our

Results: SBP was not significantly different amongst the four groups. The vasodi- 11-07 latory response to acetylcholine was significantly lower in ace2-/y mice compared Cardiorenal anemia syndrome in chronic heart failure contributes to to WT (maximal relaxation in response to acetylcholine, mean Æ SEM, ace2-/y: increased sympathetic nerve activity 85 Æ 2.2 %, vs. WT: 97 Æ 1.3 %, P < 0.001). Ang II infusion induced a signifi- F Despasa, N Franchittoa, M Labruneeb, A Vaccarob, M Galiniera, JM Senardb, cant decrease in the vasodilatory response to acetylcholine in WT mice by 25% A Pathakb aCHU Toulouse, Toulouse, France; bINSERM 1048, Toulouse, France (P < 0.05) and ace2-/y mice by 28% (P < 0.05), compared to control mice. In Background: We sought to assess whether cardiorenal anemia syndrome ace2-/y mice, Ang II-induced endothelial dysfunction was blunted by the presence (CRAS) in chronic heart failure (CHF) patients contributes to sympathetic overac- of Ang-(1-7) in the myograph chamber, an effect not observed in WT mice. No tivity through modulation of sympathetic reflexes. significant difference was observed in vascular remodeling between the four Methods and results: We prospectively studied 15 patients with CRAS and CHF groups. and 15 control CHF patients, matched for age, gender distribution, type of cardio- Conclusion: These results indicate that genetic absence of ACE2 causes endothe- myopathy, left ventricular ejection fraction (LVEF) and BMI. We compared muscle lial dysfunction, independent of changes in blood pressure. The data suggest that sympathetic nerve activity (MSNA) and the effect of peripheral chemoreflex deacti- endogenous ACE2 exerts a protective effect on resistance artery endothelial func- vation on MSNA in both groups. We also compared sympathetic baroreflex function, which may be partly mediated by production of Ang-(1-7). tion, assessed by the slope of the relationship between MSNA and diastolic blood Keywords: resistance artery, endothelial function, angiotensin-converting pressure in both groups and while peripheral chemoreflexes were (by breathing enzyme 2. 100% oxygen for 15 min) or not deactivated. Baseline MSNA was significantly elevated in CHF patients with CRAS compared with control CHF patients (83.1 Æ 4.6 vs. 64.9 Æ 2.9 bursts/100 heart beats; P < 0.05) and sympathetic baroreflex 11-05 impaired (2.69 Æ 0.44 vs. 5.25 Æ 0.60 % bursts/mmHg; P < 0.01). Chemoreflex Involvement of cytochrome epoxygenase metabolites in cutaneous deactivation with administration of 100% oxygen led to a significant decrease in postocclusive hyperemia in humans muscle sympathetic nerve activity (77.8 Æ 4.7 vs. 82.1 Æ 4.9 bursts/100 heart JL Cracowskia, F Gaillard-Bigota, C Cracowskia, C Sorsa, M Roustita, beats; P < 0.01) and to an increase in sympathetic baroreflex function C Milleta aUnite de Pharmacologie Clinique, Inserm CIC3, Grenoble, Grenoble, France (2.77 Æ 0.45 vs. 5.63 Æ 0.73 % bursts/mmHg; P < 0.01) in patients with CRAS Objective: Several mediators contribute to postocclusive reactive hyperemia and CHF. In contrast, neither room air nor 100% oxygen changed MSNA, hemody- (PORH) of the skin including sensory nerves and endothelium derived namic or sympathetic baroreflex function in control CHF patients. hyperpolarizing factors (EDHF). The main objective of our study was to investi- Conclusions: CRAS in CHF patient is associated with elevated sympathetic activ- gate the specific contribution of epoxyeicosatrienoic acids in human skin ity mediated by both tonic activation of peripheral chemoreflex and baroreflex PORH. impairment. Methods: Eight healthy volunteers were enrolled in two placebo controlled Keywords: Sympathetic nervous system, heart failure, anemia, renal failure, experiments. In the first experiment we studied the separate and combined effects baroreflex, chemoreflex. of 6.5 mM fluconazole, infused through microdialysis fibers, and lidoca€ıne/prilo- ca€ıne cream on skin PORH following 5 min arterial occlusion. In the second experiment we studied the separate and combined effects of 6.5 mM fluconazole 11-08 and 10 mM L-NMMA. Skin blood flux was recorded using two-dimensional Laser Electromyostimulation decreases muscle sympathetic nerve activity in Speckle Contrast Imaging. Maximal CVC was obtained following 29 mM sodium patients with chronic heart failure nitroprusside perfusion. M Labruneea, F Despasa, N Franchittoa, P Marquea, T Guiraudb, A Vaccarob, a a a b Results: The PORH peak at the placebo site averaged 66 Æ 11 %CVCmax. Com- M Galinier , JM Senard CHU Toulouse, Toulouse, France; INSERM 1048, pared to the placebo site, the peak was significantly lower at the fluconazole Toulouse, France (47 Æ 10 %CVCmax; P < 0.001); lidoca€ıne (29 Æ 10 %CVCmax; P < 0.001); and Purpose: Muscle passive contraction of lower limb by NeuroMuscular ElectroSti- fluconazole + lidoca€ıne (30 Æ 10 %CVCmax; P < 0.001) sites. The effect of fluco- mulation (NMES) is a validate tool to improve health status of Chronic heart fail- nazole on the area under the curve was more pronounced. In the second experi- ure (CHF) patients but no data are available concerning its action on ment, the PORH peak was significantly lower at the fluconazole site, but not at sympathetic activity. However, Transcutaneous Electrical Nerve Stimulation the L-NMMA or combination site, compared to the placebo site. (TENS) is able to improve baroreflex in CHF. The primary aim of the present Conclusions: In addition to sensory nerves cytochrome epoxygenase metabo- study was to investigate the acute effect of TENS and NMES compared to Sham lites, putatively epoxyeicosatrienoic acids, play a major role in healthy skin stimulation on sympathetic overactivity as assessed by MSNA. PORH, their role being more important in the time course rather than the peak. Methods: We performed a randomized double blinded crossover sham controlled ClinicalTrials.gov Identifier: NCT01290198. study in 22 CHF patients in NYHA III (58.5 Æ 2.6 years, LVEF 26.6 Æ 1.9 %, Keywords: epoxyeicosatrienoic acids, cytochrome P450; nitric oxide, endothe- ischemic cardiopathy cause in 64%). Half of them performed sensory stimulation lium, reactive hyperemia, microdialysis, skin, microcirculation, Laser Speckle by TENS (protocol A), and the others tested NMES stimulation (protocol B). In Contrast Imaging. each protocol, Sham stimulation corresponding to light intermittent sensory stimulation served as placebo. MSNA, Blood pressure, Heart beat were monitored during the procedures. 11-06 Results: Compare to Sham stimulation, Both TENS and NMES are able to reduce S-nitrosoglutathione abolishes angiotensin II vasoconstriction effect on MSNA (63.5 Æ 3.5 vs. 69.7 Æ 3.1 burst/min, P < 0.01 after TENS and middle cerebral artery: possible link with nitrosation of AT1 receptor 51.6 Æ 3.3 vs. 56.7 Æ 3.3 burst/min, P < 0, 01 after NMES). No variation of A Markovetsa, J Metzgera, C Perrin-Sarradoa, C Gauchera, I Lartauda, blood pressure or heart rate was observed after stimulation. F Dupuisa aUniversite de Lorraine, Nancy, France Conclusions: The results suggest that sensory stimulation of lower limbs by Objective: Nitric oxide (•NO) and renin-angiotensin-aldosterone system (RAAS) electrical device either TENS or NMES could inhibit sympathetic outflow directed play a critical role in arterial pressure as in cerebral blood flow regulation [1]. to legs in CHF patients. These properties could explain beneficial effects of NMES Many of the biological effects of •NO are mediated by the direct modification of in CHF besides its known muscular strengthening. cysteine residues in proteins by S-nitrosation. S-nitrosation of the angiotensin II Keywords: Sympathetic nervous system, heart failure, electromyostimulation. (AngII) type 1 receptors (AT1R, responsible for the vasoconstrictive effect of AngII) alters its affinity for AngII [2]. We studied the functional impact of S-nitrosation of angiotensin receptors at the cerebrovascular level, using S-nitro- 11-09 soglutathione – GSNO – to produce nitrosation. Mortality and major cardiovascular morbi-mortality 3.5 years after an Methods: Isolated middle cerebral arteries (MCA) from male adult Wistar rats acute myocardial infarction from the EOLE real-life cohort study were mounted in a small vessel arteriograph and pressurized (80 mmHg), contin- C Droza, C Dureaub, D Thomasc, N Danchind, J Tricoiree,JBenichouf, F Paillardg, uously perfused (100 lL/min) and immersed in a physiologic saline solution S Hercbergh, I Siboni, F Rouaneti, S Rambelomananab,HMa€ızib, MA Bernardb,P b j a (PSS). After 1 h equilibration, KCl (40 mM), then concentration response curves Blin , N Moore INSERM CIC-P 0005, INSERM U657, Universite de Bordeaux, À13 À6 b to AngII, serotonin or GSNO (10 –10 M) were built to determine EC50 of Bordeaux, France; INSERM CIC-P 0005, Universite de Bordeaux, Bordeaux, France; the internal diameter responses. To study the impact of S-nitrosation, MCA were cHôpital Pitie-Salpêtriere, Paris, France; dHôpital Europeen Georges Pompidou, Paris, e f exposed for 30 min to GSNO (2 lM), followed by 1 h washing with PSS [3], then France; Cardiologue, Toulouse, France; CHU de Rouen, INSERM U657, Rouen, AngII and serotonin were administrated at equiactive concentrations. Results are France; gCHU de Pontchaillou, Rennes, France; hINSERM U557, Bobigny, France; presented as percent of the internal diameter measured after equilibration (mean iCHU de Bordeaux, Bordeaux, France; jINSERM CIC-P 0005, INSERM U657, CHU esm) and compared with a one-way ANOVA + post-hoc Neumann-Keuls (P value < de Bordeaux, Universite de Bordeaux, Bordeaux, France 0.05). Objective: Interim analysis to estimate the event rate after a median follow-up Results: Concentration response curves for AngII (Emax À15 Æ 5%, logEC50 of 3.5 years for a study aiming to assess the impact of recommended secondary 10.2 Æ 0.4, n = 9), serotonin (Emax À46 Æ 3%, logEC50 7.5 Æ 0.3, n = 3) and prevention drugs on all-cause mortality and major cardiovascular morbi-mortal- GSNO (Emax +43 Æ 7%, logEC50 5.6 Æ 0.2, n = 5) allowed to determine the ity 6 years after an acute myocardial infarction. À9 equiactive concentrations for AngII (3.10 M, control À8.6 Æ 0.3%) and seroto- Methods: A cohort study was designed to include 5000 patients with recent À9 nin (3.10 M, control À11 Æ 1%) (n = 7, P > 0.120). After exposure to GSNO acute myocardial infarction (<3 months) recruited at or after hospital discharged (n = 7), the response to AngII was abolished (-0.4 Æ 0.5%, compared to control by hospital and non-hospital cardiologists. Patients were followed-up for 6 years without GSNO) and significantly lower than that of serotonin (10 Æ 1%). with a yearly self-administered questionnaire to collect current treatment, hospi- Conclusions: Pretreatment with GSNO abolished the vasoconstriction effect of talisations, and death. Deaths were also searched from patients’ family, cardiolo- AngII but not that of serotonin. Involvement of AT receptors nitrosation have to gists and general practitioners for patients lost to follow-up, as well as from the be investigated on suitable models. National Death Registry for the whole cohort at 3.5 years of follow-up. Declared Acknowledments: The work was funded by the French National Research hospitalisations were investigated to obtain discharge summaries, and deaths Agency (ANR NanoSNO 2010, Pr. P Leroy, EA3452 CITHEFOR). were investigated to ascertain the cause. An independent event committee, References: blinded to drug exposure, validated all cardiovascular events and causes of death. 1. Foulquier et al. PLoS One (2012) 7(9) e42469. The Kaplan Meier and the patient-year methods were used to estimate mortality 2. Leclerc et al. Br. J. Pharmacol. (2006) 148(3) 306–313. and morbi-mortality. 3. Sarr et al. Nitric Oxide (2007) 17(1) 1–9. Results: Between May 2006 and June 2009, 5538 patients were included. Dur- Keywords: Angiotensin II, S-nitrosoglutathione, isolated middle cerebral artery. ing follow-up, 7239 hospitalisations or deaths were declared, 2475 cardiovascular events and causes of death were examined by the Event Validation

Committee, with 1799 major cardiovascular events and causes of death vali- 11-12 dated, including 59 deaths registered only by the National Death Registry. After Deletion of the mineralocorticoid receptor in vascular smooth muscle 3.5 years of follow-up, the event rate was 7.8% (95%CI: 7.1–8.5) for all-cause cells improves coronary and cardiac function in mice with heart failure mortality, 1.9% (1.6–2.3) for coronary mortality, 2.5% (2.1–2.9) for cardiovascu- A Guereta, G Galmicheb, L Nicola, N Haroukia, JP Henrya, M Besniera,C lar mortality, 14.8% (13.8–15.8) for major coronary morbi-mortality, and 23.1% Thuilleza, V Richarda, P Muldera, F Jaisserb, A Ouvrard Pascauda aInserm u1096, (21.9–24.3) for major cardiovascular morbi-mortality. The incidence for 1000 Rouen, France; bInserm u872, Paris VI, France patient-years of follow-up was 23.2 for all cause death, 5.6 for coronary deaths, The RALES, EPHESUS and EMPHASIS clinical studies proved the efficacy of min- 7.3 for cardiovascular deaths, 50.8 for major coronary events, and 83.1 for eralocorticoid receptor (MR) antagonists in chronic heart failure (CHF). Recently major cardiovascular events. Fraccarolo et al. showed that MR deletion in cardiomyocytes improves cardiac Discussion: There are few recent large and long-term real-life follow-up studies remodeling and function in CHF, however the specific role of vascular MR in this after myocardial infarction, especially in France. This interim analysis shows that disease remains unknown. Thus, the vascular and cardiac consequences of MR the mortality and major cardiovascular morbi-mortality rates after a median of deletion specifically in vascular smooth muscle cells (VSMCMRÀ/À) were studied 3.5 years of follow-up are within the study hypothesis to assess the impact of the in mice with CHF (2 months coronary artery ligation). recommended secondary prevention treatment 6 years after an acute myocardial Invasive left ventricular (LV) pressure-volume curves showed that VSMCMRÀ/À infarction. CHF mice had increased LV end-systolic pressure-volume relationships (elastance; Keywords: Acute myocardial infarction, mortality, cardiovascular morbi- ctrl 11.9 Æ 0.5, n = 7; VSMCMRÀ/À 19.9 Æ 2.5, n = 6, P < 0.05) and decreased mortality. LV end-diastolic pressure-volume relationships (compliance; ctrl 4.08 Æ 0.45; VSMCMRÀ/À 2.16 Æ 0.33, P < 0.01), in the absence of changes in cardiac fibrosis (% collagen density ctrl 1.23 Æ 0.33, n = 5; VSMCMRÀ/À 1.40 Æ 0.31, n = 4, 11-10 ns). The CO-releasing molecule, CORM-3, protects adult cardiomyocytes Coronary endothelial dysfunction was assessed ex vivo in isolated coronary seg- + ments mounted on a small vessel myograph, by testing the NO-mediated response against hypoxia-reoxygenation by inhibiting Na /HCO3- symporter MRÀ/À L Portala, B Ghaleha, A Berdeauxa, R Motterlinia, S Ponsa aINSERM U 955 to acetylcholine. In sham-operated animals, no difference between VSMC mice and littermate controls were observed (maximal relaxation ctrl 91 Æ 1, Equipe 03, Universite Paris Est Creteil, Creteil, France MRÀ/À Objectives: Several studies have reported that CORM-3, a water-soluble carbon n = 5; VSMC 83 Æ 2%, n = 4, ns). CHF was associated with reduced response to acetylcholine, indicating endothelial dysfunction, which was less monoxide (CO)-releasing molecule, elicits cardioprotection against myocardial MRÀ/À MRÀ/À infarction but the mechanism of its beneficial action remains to be elucidated. severe in VSMC mice (ctrl 30 Æ 3, n = 4; VSMC 54 Æ 2%, n = 4, Considerable evidence also indicates that inhibition of two pH regulators, the P < 0.01). + + + À Finally, coronary reserve was measured by magnetic resonance imaging as the Na /H exchanger (NHE) and Na /HCO3 symporter (NBS), protects cardiomyocytes from hypoxia/reoxygenation injury by delaying the intracellular pH (pHi) difference between basal and maximal LV perfusion obtained after stimulation by recovery at the time of reperfusion. Thus the aim of this study was to explore the A2a adenosinergic agonist ATL307. In sham-operated mice, MR deletion in whether CORM-3-mediated cytoprotection involves the modulation of NBS and VSMC resulted in a higher coronary reserve, although this did not reach statistical significance probably due to the small size of the groups in these preliminary other well-known cardioprotective targets such as mitochondrial ATP-dependent MRÀ/À K+ channels (mK ) and ERK ½ pathway. data (ctrl 3.4 Æ 0.9, n = 4; VSMC 6.7 Æ 1.4 mL/mg/min, n = 4, ATP P = 0.10). In CHF mice, VSMC MR deficiency was associated with an increased Materials: Cardiomyocytes freshly isolated from adult mice (C57BL6J) were sub- MRÀ/À mitted to a sequence of 3 h hypoxia followed by 2 h of reoxygenation. CORM-3 basal coronary perfusion (ctrl 10.5 Æ 0.41; VSMC 12.0 Æ 0.5 mL/mg/ l min, P < 0.05), probably responsible for the observed decrease in coronary (20, 50 and 100 M) was added at the time of reoxygenation and cell viability MRÀ/À was assessed using trypan blue. reserve (ctrl: 4.5 Æ 0.7, n = 12; VSMC 4.5 Æ 0.7 mL/mg/min, n = 9, Results: NBS activity was modulated using different experimental conditions: 1) P < 0.01). medium with or without bicarbonate since NBS is inhibited in bicarbonate-free These results suggest that MR deletion in VSMC improves coronary endothelial medium; 2) hypoxia performed with extracellular pH (pHo) either at 7.4 or 6.2, function and cardiac perfusion, and this is associated with reduced cardiac dys- followed by a reoxygenation phase at pHo 7.4. The contribution of NBS is of function in this model of CHF. major importance for acid-extrusion when pHi at the end of hypoxia is close to Keywords: mineralocorticoid receptor, vascular smooth muscle cell, heart neutrality but is rather low when pHi is acidic. CORM-3 reduced by an average failure. of 15% (P < 0.05, n = 15) the mortality of cardiomyocytes placed in bicarbonate-buffered solution and exposed to hypoxia at pHo 7.4. Under acidic conditions during hypoxia, the extent of cardioprotection was weaker. Interestingly, the cardioprotective effect of CORM-3 was abolished by switching to a bicarbonate-free SESSION 12: SUIVI THERAPEUTIQUE, medium (i.e. NBS inhibition), independently from pHo used during hypoxia. Fur- thermore, the beneficial effect of CORM-3 was also inhibited by 5-hydroxydecano- PHARMACOLOGIQUE, PHARMACOCINETIQUE, l ½ l ate (mKATP inhibitor, 500 M) or PD098059 (ERK inhibitor, 10 M). PHARMACOGEN ETIQUE Conclusion: mKATP channels opening, activation of ERK ½ pathway and inhibi- + À tion of Na /HCO3 symporter at reoxygenation contribute to the cardioprotection conferred by CORM-3. 12-01 Keywords: CO-releasing molecule, hypoxia, cardiomyocyte. The effect of Roux-en-Y gastric bypass on oral morphine pharmacokinetics C Lloret-Linaresa, C Bardinb, D Hirtc, C Poitoud, JL Bouillote, X Declevesc,S 11-11 Moulyc,f, JF Bergmannc,f aAssistance Publique-Hôpitaux de Paris, Service de Medecine Number of CD14+ cells is related to infarct size and postinfarct volumes Interne A, Unite de recherche therapeutique-Hôpital Lariboisiere-75475 Paris Cedex in ST segment elevation myocardial infarction: a monocentric clinical 10, Universite Paris VII, Paris Cedex 10, France; bAssistance Publique-Hôpitaux de study Paris, Department of Toxicology, Hôpital Lariboisiere, Paris F-75010, France, Paris, D Montangea, S Davania, N Meneveaub aLaboratoire de Pharmacologie Clinique et c b France; Universite Paris Cite-Descartes, Faculte de pharmacie, Unite INSERM U705, Toxicologie – CHRU de Besancßon, Besancßon, France; EA 3920 – Universitede CNRS UMR 7157, 75006-Paris, Paris, France; dAssistance Publique-Hôpitaux de Franche Comte, Besancßon, France Paris, Universite Paris VI, Service de Nutrition-Hôpital La Pitie Salpetriere-Paris, Objective: Left ventricular (LV) remodeling after acute myocardial infarction Paris, France; eAssistance Publique-Hôpitaux de Paris, Service de Chirurgie viscerale, (MI) is characterized by increasing LV volumes, which result in impairment of Hôpital Ambroise Pare-Boulogne, Universite Paris-Ouest, Boulogne, France; fAssistance myocardial function. Many factors influence LV remodeling, particularly infarct Publique-Hôpitaux de Paris, Service de Medecine Interne A, Unite de recherche location and the extent of necrosis. Acute MI is associated with an increased therapeutique-Hôpital Lariboisiere-75475 Paris Cedex 10, Universite Paris VII, Paris release of monocytes into the bloodstream. These monocytes contribute to ische- Cedex 10, France mic tissue healing after acute MI, which should help to prevent LV remodeling. Objectives: Obese patients are at high risk of several chronic pains that may The objectives of this clinical study were to determine the relationship between require morphine. Our objective was to describe the changes in oral morphine the number of CD14+ cells, MI size and LV volumes in ST segment elevation MI pharmacokinetics after drastic alteration in gastrointestinal anatomy resulting (STEMI) and non-ST segment elevation MI (NSTEMI) patients. from Roux-en-Y gastric bypass (RYGB). Methods: A total of 62 patients (younger than 75 years) with STEMI (n = 34) Methods: Oral morphine (Oramorphâ, 30 mg, 5 mL) pharmacokinetics was or NSTEMI (n = 28) were enrolled. Peripheral blood samples were drawn on determined before (n = 31), immediately after RYGB (in the 15 following days, admission after the onset of MI. The number of CD14+ cells was assessed with a n = 25) and 6 months after RYGB (n = 26) in obese volunteers [Age: 41 (18– FACsort flow cytometer. Infarct size, left ventricular ejection fraction (LVEF) and 62) years, BMI : 44.6 (35.4–62.2) kg/m2]. Plasma concentrations of morphine LV volumes were measured using magnetic resonance imaging 5 days after MI were quantified using a validated ion-pair reverse phase HPLC assay with electro- and 6 months after MI. chemical detection. A non-compartmental pharmacokinetics analysis was per- Results: No significant difference was observed between NSTEMI and STEMI formed using the WINONLIN software (Pharsight, USA). groups in term of age, cardiovascular risk factors and previous medication. In Results: The median Cmax (lg/L) was 11.3 (6.4–24.0) before surgery, 19.1 STEMI patients, the number of CD14+ cells was positively and significantly corre- (9.0–38.3) immediately after and 38.1 (16.7–77.2) 6 months after; the median lated with infarct size at day 5 (r = 0.40; P = 0.016) and after 6 months Tmax (min) was 53 (31–84), 23 (12–37) and 7 (3–11) respectively whereas (r = 0.34; P = 0.047), negatively correlated with LVEF at day 5 (r = À0.50; AUC0-infinity (lg/L h) was 44.8 (20.1–68.5), 52.5 (27.6–77.4) and 54.7 (25.4– P = 0.002) and after 6 months (r = À0.46; P = 0.005) and positively correlated 113). with end-diastolic (r = 0.38, P = 0.02) and end-systolic (r = 0.49, P = 0.002) Discussion: Morphine absorption is fourfold increased immediately after surgery volumes after 6 months. In NSTEMI patients, no significant correlation was and 20-fold increased at 6 months, whereas exposition is moderately increased. found between the number of CD14+ cells and infarct size, LVEF or LV volumes Consequences of these pharmacokinetic changes may be important in the clinical at day 5 or after 6 months. practice. Oral morphine must be prescribed carefully after RYGB. Conclusion: The number of CD14+ cells at admission was associated with Keywords: Morphine, bariatric surgery, obesity, pharmacokinetics. infarct size and LV remodeling in STEMI patients with large infarct size, whereas in NSTEMI patients, no relationship was observed between numbers of CD14+ cells and LV remodeling. Keywords: monocytes, myocardial infarction, remodelling.

12-02 Therapeutic Drug Monitoring (TDM) of the main active metabolite of ABZ, ABZ Investigation of drug-drug interaction between dabigatran and sulphoxyde (ABZ-SO) plasmatic levels are recommended. However, no data are claritromycine available on the pharmacokinetics of ABZ-SO in liver lesion due to alveolar echi- X Delavennea, E Olliera, T Basseta, S Laportea, P Mismettia aCHU de Saint-Etienne, nococcosis. Our prospective study aimed to provide data on intralesionnal ABZ- Saint-Etienne, France SO concentrations in patients with liver alveolar echinococcosis. Background: Dabigatran etexilate (DE), an oral direct thrombin inhibitor, is Methods: AE patients with ABZ treatment and who benefited of surgical removal used for the prevention of thromboembolism in patients having undergone ortho- of AE lesion over a 5-year period (2008–2012) were included in this study. ABZ- paedic surgery and for the long-term prevention of stroke in patients with atrial SO concentrations were determined in tissue (non-infected liver tissue and AE fibrillation. DE is a substrate for P-glycoprotein (P-gp), an efflux transporter, lesion) and in plasma taken during surgery. ABZ-SO determinations were realized which limits its intestinal absorption. It has been shown that strong inhibitors or using a validated photodiode array-HPLC method. Clinical, radiological, serologi- inducers of P-gp modified the bioavailability of DE. cal, and pathological data were prospectively collected for each patient. Objective: The aim of this study was to develop a semi mechanistic PK/PD Results: Twelve patients with AE liver resection were included in this study. model to assess drug-drug interactions between dabigatran caused by P-gp modu- Operative specimens were sampled 10–19 h after last ABZ intake (median time: lation taking the example of clarithromycin, a strong inhibitor of P-gp. 16 h). Mean [min; max] ABZ-SO concentrations were respectively of 0.77 lM Subjects and methods: Each subject (n = 10) received at the first period a sin- [0.21; 3.47] in per-operatively-collected plasma. ABZ-SO were significantly higher gle 300 mg dose of DE and in the second treatment period 500 mg of clarithro- in AE liver lesion with a mean concentration at 1.52 nmol/g [<0.10; 8.97] than mycin twice daily during 3 days and then 300 mg of DE concomitantly with in non-infected liver tissue with a mean concentration at 0.42 nmol/g (<0.10; 500 mg of clarithromycin on the 4th day. Dabigatran plasma concentration and 1.6; P = 0.025 Wilcoxon test). ecarin clotting time (ECT) were measured. Modelling was performed in two steps, Discussion: Our study provides for the first time observations of diffusion of focusing first on pharmacokinetics (PK), then on PK and pharmacodynamics ABZ-SO into AE lesion in the liver, with significant higher ABZ-SO concentrations (PD). Models were built using a non-linear mixed effect modelling approach with observed in AE lesion vs. non-infected liver tissue. This fact might be explained MONOLIX software. by accumulation and/or lower clearance of ABZ-SO from the lesion. Further stud- Results: The PK/PD model was based on an inverse Gaussian absorption process ies are needed, focusing on potential relationship between the type of lesion and to describe the absorption process with two compartments. The relationship intralesional ABZ-SO diffusion. between dabigatran concentration and ECT was implemented as a linear func- Keywords: albendazole; alveolar echinococcosis; tissue concentrations. tion. The coadministration of clarithromycin induced a significant change only in DE bioavailability parameter, which increased from 6.5% to 10.1%. Exposure to dabigatran (AUC) and ECT were increased by nearly 50% with concomitant 12-05 administration of clarithromycin. A general model for quantitative prediction of drug exposure in case of Conclusion: This population model study well demonstrates the drug-drug inter- drug-drug interactions on polymorphic cytochroms 2D6, 2C9 and 2C19 actions between dabigatran and clarithromycin by an increase of bioavailibility. M Toda, S Goutellea, C Castellanb aFaculte de Pharmacie Lyon 1, Lyon, France; This model could be used as an efficient tool to investigate other drug-drug inter- bFacultedemedecine Lyon est, Lyon, France actions with dabigatran implying p-gp inhibitors, inducers or substrates. Background: A new framework to predict quantitatively the impact of CYP Keywords: drug-drug interaction, dabigatran, clarithromycin, P-glycoprotein polymorphism and drug-drug interactions (DDI) on drug exposure is proposed. inhibition, bioavailability, population pharmacokinetics and pharmacodynamics. The metrics of interest is the ratio of victim drug AUC when the inhibitor is present, to the AUC of victim drug administered alone. This ratio may be calculated for any genotype of a polymorphic cytochrom. The model relies only on in vivo 12-03 data, avoiding extrapolations, and uses several parameters: A polymorphism in MRP4 influences the decrease in neutrophils induced • The fraction of victim drug metabolized by the CYPs in extensive metabolizers. by ganciclovir in a cohort of renal transplant recipients • The fraction of activity of the mutated allele combination with respect to the PA Billata, F Saint-Marcouxb, JB Woillardb, P Mervillec, L Rostaingd, N Kamard, wild homozygous. JP Rerollee, M Essige, P Marquetb, N Picarda aInserm UMR S-850, Laboratoire de • The inhibition ratio of the inhibitor for the CYPs involved in victim drug Pharmacologie medicale, Facultedemedecine, Limoges, France; bInserm UMR S-850, metabolism. Service de Pharmacologie, Toxicologie et Pharmacovigilance, CHU Limoges, Limoges, Aim: Using values of the parameters previously established by our group, the France; cHôpital Pellegrin, CHU Bordeaux, Service de Nephrologie, Transplantation, aim was to evaluate the model by external validation by comparing predicted Dialyse, Bordeaux, France; dCHU Toulouse Rangueil, Service de Nephrologie, HTA, AUC ratios to published values for a wide range of situations. Dialyse et Transplantation d’Organes, Toulouse, France; eInserm UMR S-850, Service Methods: An exhaustive litterature search was carried out to extract the AUC de Nephrologie, CHU Limoges, Limoges, France ratios measured for PK interactions on CYP2D6, 2C9, 2C19 and 3A4 in patients Background: Cytomegalovirus (CMV) infection is a major issue in transplanta- with various genotypes. Three formulaes were derived to accomodate different tion, being associated with a high morbidity. Prophylactic or pre-emptive therapy cases. with ganciclovir (GCV) is used in this context, but it induces hematological Results: Data were available for 12 drugs, 27 DDIs, with 67 AUC ratios mea- adverse effects (mainly neutropenia) leading to premature treatment discontinua- sured. All predictions were in the range of 50–200% of the observed values. The tions or to the use of lower doses, favoring the emergence of resistance. mean prediction error, mean absolute error and mean relative absolute error of Objective: We aimed at identifyingpolymorphisms in the genes encoding GCV the AUC ratios were 0.03%, 0.31% and 19% respectively. membrane transporters linked to drug toxicity in a cohort of renal transplant Discussion: The model is unbiased and sufficiently precise for prediction of drug patients. exposure under DDI in patients with various CYP2D6, 2C9, and 2C19 genotypes. Methods: Genomic DNA was obtained from 207 patients enrolled in the Epigren For a drug metabolized by e.g. 2D6 and 3A4, and a DDI mediated by 2D6, the study, an on-going cohort of renal transplant patients recruited over a 4-year per- AUC ratio is lowest in poor metabolizers and greatest in ultrametabolizers. The iod. The selection of genes and polymorphisms was based on the following criteria: reverse order prevails if the DDI is mediated by 3A4. Clinical consequences of (i) transporters expressed in blood cells; (ii) in vitro evidence of purine analogue DDIs depend strongly on patient genotype when 2D6, 2C9 or 2C19 are involved. (ideally GCV) transport; and (iii) clinical evidence of polymorphism functionality. Keywords: drug interactions, cytochrom, polymorphism, mathematical model. Genotyping was performed using pre-designed or custom Taqmanâ genotyping assays (Applied-Biosystems). Statistical analysis of associations between genotypes, co-treatments, and the evolution of neutrophil counts after drug introduction was 12-06 performed using multivariate, linear mixed-effect modeling with the R program. An innovative model of time-dependent methotrexate elimination Results: Fourteen polymorphisms in seven genes (SLC29A1, SLC28A3, JB Woillarda,b, J Deborda,b, M Neelyc, P Marqueta,b, F Saint-Marcouxa,b aCHU de SLC28A2, SLC22A1, ABCG2, ABCC4 and ABCB1) were selected. The decrease in LIMOGES, Service de Pharmacologie, Toxicologie et Pharmacovigilance, Limoges, neutrophil count was more pronounced in patients carrying the ABCC4 (MRP4) France; bINSERM UMR-S850, Univ. Limoges, Limoges, France; cLaboratory of applied rs11568658 variant allele (n = 7) than in non-carriers (n = 157) (P = 0.0004). Pharmacokinetics, University of Southern California, Los Angeles, CA, USA No significant association with other polymorphisms (P > 0.10), co-treatments Objective: Methotrexate (MTX) therapeutic drug monitoring (TDM) is performed [i.e. trimethoprim (P = 0.12) or mycophenolate mofetil (P = 0.30)] and GCV dose to determine when MTX concentration falls below a predefined threshold, at (P = 0.08) was found. which time leucovorin rescue therapy can be withdrawn. Existing pharmacoki- Conclusion: This study suggests thatMRP4 rs11568658 could be associated netic (PK) models predict MTX clearance in a given patient, but none account for with a higher risk of neutropenia in transplant patients given GCV. This SNP, time-dependent changes in MTX clearance after an infusion. In this study, we known to decrease MRP4 activity [1], might favor GCV accumulation in neu- developed a Bayesian model that included time-dependent MTX elimination. trophils. This finding is consistent with previous in vitro evidence that GCV is a Materiel and methods: We obtained 105 MTX PK profiles from patients at the substrate for MRP4 [2]. Transfected cellular models are being set up to confirm Limoges Hospital University with 499 concentrations sampled up to 113 h after this hypothesis. the end of infusion. We used 70 profiles for model building and 35 for model val- References: idation. To estimate model parameter values in the population, we used the non- 1. Abla et al. J. Pharmacol. Exp. Ther. (2008). parametric adaptive grid algorithm in the Pmetrics R package. We defined MTX 2. Adachi et al. J. Biol. Chem. (2002). terminal elimination k(t) as a power function: k(t) = a/1+bt. Keywords: Cytomegalovirus, Ganciclovir, Neutropenia, Gene polymorphism, Pro- Results: The model accurately fitted the data with a mean (SD) relative bias of phylaxis. 0.019 Æ 0.1313 (RMSE = 13.2%) between observed and predicted concentrations. Bayesian posterior estimates of model parameter values in the validation group yielded good concentration estimation (relative bias of 0.017 Æ 0.1381; 12-04 RMSE = 13.9%). The threshold of 0.05 mg/L was reached after 36.0 (34.5– Determination of tissue albendazole sulphoxide concentrations in 48.0) h after the dose, and the difference between the observed and predicted patients with alveolar echinococcosis times was 2.71 (0.70–3.47) h. D Montangea, F Grenouilletb, MP Brientinia, S Piedouxa, B Royera, P Mureta, Discussion: We developed a model with post-dose time-dependent elimination S Davania aLaboratoire de Pharmacologie Clinique et Toxicologie – CHRU de for MTX, which predicts the time when leucovorin therapy may be discontinued Besancßon, Besancßon, France; bLaboratoire de Parasitologie – CHRU de Besancßon, 3 Bd more accurately and precisely than previous models. The model is easily general- Fleming, Besancßon, France izable to other drugs, such as aminoglycosides and glycopeptides. Background: Albendazole (ABZ) therapy, alone or in combination with surgery, Keywords: Methotrexate, therapeutic drug monitoring, modelling, pharmacoki- remains the standard treatment for human alveolar echinococcosis (AE). Regular netic.

12-07 *10 and *41) variants were detected by DNA sequencing, *5 variant and dupli- Pharmacokinetic assessment of new vancomycin dosing schedule in cated alleles *1xN, *2xN by long range PCR. maintenance hemodialysis patients Results: Fifty-two patients, mean age 37 (19–86), were included over a E Chasseuila, F Darrouzaina, L Ghouti-Terkib, N Rabotb, D Ternantc,N 6 months period. For 46 patients schizophrenia was confirmed. The six lasting Azzopardic, B Birmeleb, G Paintaudc aLaboratoire de Pharmacologie-Toxicologie, ones were diagnosed as depression and generalized anxiety disorder. Mean dura- Tours, France; bService d’hemodialyse, Tours, France; cCNRS UMR 7292 GICC, tions of illness and treatment were respectively 15 (3–66) and 13 (2–56) years. Tours, France Aripiprazole, chlorpromazine, haloperidol, risperidone and olanzapine were the Objectives: Patients on maintenance hemodialysis are at a greater risk for bacte- most frequently used CYP2D6 metabolized drugs. rial infection, especially with Staphylococcus aureus. Vancomycin is commonly CYP2D6 genotyped-predicted phenotypes identified 3.8% ultrarapid, 9.6% poor used to treat these infections. Following recent guidelines, vancomycin adminis- (all of them homozygous *4/*4 except one *4Xn/*9), 28% intermediate and tration procedure was modified: instead of being administered at the end of dialy- 57% extensive metabolizers. Among the 6-revised diagnosis, two were expected sis sessions, vancomycin infusion is now started during the last hour of dialysis. to be poor, and one to be ultrarapid metabolizers. The aim of our work was to assess vancomycin exposition of maintenance dialy- A significative difference was observed in the waiting period to obtain an effective sis patients obtained by this new vancomycin dosing schedule. and safe treatment as intermediate and poor metabolizers needed respectively 2.4 Patients and method: Vancomycin serum concentrations were measured dur- and 3.6 times as long as extensive and ultrarapid metabolizers (6.3 and 9.6 vs. ing 35 sessions of dialysis performed in 13 maintenance hemodialysis patients. 2.6 and 3 years). Five blood samples were collected before dialysis session, 2 and 3 h after dialysis Difference in side effects was also significative: respectively 30, 50 and 100% of start, at the end of and 2 h after dialysis session to measure vancomycin serum extensive, intermediate and poor metabolizers experienced side effects with concentrations. Vancomycin exposition was estimated by the area under the con- CYP2D6 metabolized drugs. None of ultrarapid metabolizers reported adverse centration vs. time curve over 24 h (AUC 0–24 h) using the trapezoidal method, response. both on days including a dialysis session (AUC 0–24 h dialysis) and on days Discussion: Time to obtain ‘clinical’ steady state with CYP2D6 metabolized anti- between two dialysis sessions (AUC 0–24 h interdialysis). These AUC 0–24 h psychotic was here related to genotyped-predicted phenotypes. It could be inter- were compared to low and high theorical AUCs: AUC-L (480 mg h/L) and AUC- esting to precociously determine genotype to avoid this waiting period with H (720 mg h/L) which are the vancomycin expositions corresponding to 24 h inefficacy and side effects. We submit a prospective, randomized study to evaluate continuous infusions leading to 20 and 30 mg/L concentrations, respectively. relevance of early-coupled CYP2D6-CYP2C19 genotyping in this population. AUCs were compared using a Wilcoxon test. Keywords: CYP2D6, polymorphisms, schizophrenia, antipsychotic drugs. Results: Mean (interindividual coefficient of variation, CV) AUC 0–24 h dialysis and AUC 0–24 h interdialysis were 550 mg h/L (24.1%) and 457 mg h/L (27.7%, respectively). Mean AUC 0–24 h dialysis was therefore 13% higher than 12-10 AUC-L (P = 0.0084) but lower than AUC-H (P < 0.0001). Only six out of 35 Pharmacokinetic study of raltegravir in hiv-infected patients with end dialysis sessions led to AUC 0–24 h dialysis >720 mg h/L. stage liver disease: liveral anrs 148 study Conclusion: The new vancomycin-dosing schedule leads neither to under-expo- C Baraua, J Braunb, C Vincentb, S Haim-Boukobzac, JM Molinad, P Miailhese, sition nor to over-exposition to the antibiotic of maintenance hemodialysis JP Aboulkerb, JC Duclos-Vallef, AM Taburetg, E Teicherh aAPHP, Plateforme de patients Ressources Biologiques, Hôpital Henri Mondor, Creteil, France; bINSERM SC10- Keywords: area under the concentration vs. time curve, maintenance hemodial- US019, Villejuif, France; cAPHP, Service de Virologie, Hôpital Paul Brousse, Villejuif, ysis patients, new vancomycin dosing schedule, vancomycin serum concentra- France; dAPHP, Service des Maladies Infectieuses et Tropicales, Hôpital Saint Louis, tions. Paris, France; eHospices Civils de Lyon, Hôpital de la Croix-Rousse, Lyon, France; fAPHP, Centre Hepato-Biliaire, Hôpital Paul Brousse, Villejuif, France; gAPHP, Service de Pharmacie-Toxicologie, Hôpital Bicêtre, Le Kremlin-Bicêtre, France; hAPHP, Service 12-08 de Medecine Interne, Hôpital Bicêtre, Le Kremlin-Bicêtre, France Quantitative prediction of oral drug interactions caused by CYP3A4 Background: Raltegravir (RAL), metabolized via UDP glucuronosyltransferase inhibition for an extended panel of substrates and inhibitors 1A1, is part of antiretroviral therapy. Because of its good tolerance, studying its C Louea, M Todb aPharmacie, Hopital de la Croix-Rousse, Lyon, France; bUniversite relevance in the context of severe liver disease is crucial. The aim of this study Lyon 1, Lyon, France was to evaluate pharmacokinetic parameters of raltegravir in HIV infected Background and objectives: A framework to predict quantitatively the impact patients with end stage liver disease (ESLD). of CYP3A4 drug-drug interactions (DDI) on drug exposure (the AUC ratio) has Methods: Patients with HIV-RNA <50 cps/mL and eligible for liver transplanta- been proposed previously by Ohno (2007) for 14 substrates and 18 inhibitors. tion were included. Antiretroviral therapy was switched to RAL 400 mg bid. This approach is based on in vivo data and relies on two characteristic parame- Liver function assessed by MELD and Child-Pugh (CP) scores and immunovirolog- ters: the contribution ratio (CR: the fraction of victim drug clearance due to ical parameters were checked at J0 and up to M3 after RAL initiation. Serial metabolism by a specific CYP) and the inhibition ratio (IR) of the inhibitor. Here, blood samples were drawn at M1 after RAL initiation over a 9 h-dosing interval we extended this method to a larger set of CYP3A4 substrates and inhibitors. post RAL intake. Patients had to fast up to 1 h post RAL intake. Plasma concen- Methods: A two-step approach was used. First, initial estimates of CRs and IRs tration of total RAL, unbound RAL (RALu) and inactive RAL glucuronide (RALg) were obtained by several methods, using data from the literature. Second, an were measured by LC/MS/MS. RAL unbound fraction (fu) is the ratio of RALu to external validation of these initial estimates was carried out, by comparing the RAL. Pharmacokinetic (PK) parameters were analyzed by a model independent predicted AUC ratios with the observed values. Predictive performance was evalu- method. Metabolic ratio (MR) was calculated from the AUC ratio of RALg to ated by the mean prediction error (MPE) for bias, and the mean absolute predic- RAL. Results are presented as median (range). tion error (MAPE) for precision. Results: Ten patients (70% males, 80% VHC+) were included in this study: age: Results: Sixty-seven AUC ratios were available for the global analysis. Ten of 50 (39; 63) years, CD4: 258 (57; 604) cells/mm3. MELD and CP scores at them were excluded because the DDI had multiple mechanisms (involving trans- screening were 12 (5; 26) and 10 (6; 14) respectively. Median MELD at M1 was porters). Final estimates of CRs and IRs were obtained for 29 substrates and 22 11 (7; 33). Albumin concentration at M1 was 27 (24; 36) g/L. HIV-RNA inhibitors, respectively. Forty-four AUC ratios were available for external valida- remained <50 cps/mL at M1 and M3. RAL Ctrough, Cmax and AUC0-9 were 0.9 tion. The MPE of the ratios was – 0.169, while the MAPE was 0.99. (0.1; 18.4), 19.7 (2.0; 36.5) lM and 75.3 (11.6; 148.6) lM h respectively. Discussion: Among the drug newly characterized, strong inhibitors (IR > 0.75) Unbound Ctrough and Cmax were 0.2 (0.1; 3.0), 3.5 (0.4; 5.9) lM and 13.8 included posaconazole, telaprevir and boceprivir. Substrates at risk (CR > 0.75) (2.0; 22.2) lM h, leading to a Fu ratio of 0.19 (0.12; 0.32) not related to albu- are dronedarone, sildenafil, tacrolimus, ticagrelor and triazolam. AUC ratios for min concentrations. RALg AUC was 46.8 (10.1; 369.2) lM h and MR ratio was 638 possible interactions were obtained. These results allow extending the quan- 0.9 (0.3; 2.5). All the PK parameters analyzed did not differ from those estimated titative prediction of CYP3A4-mediated drug-drug interactions to a broader range in patients without liver disorders. of substrates and inhibitors. Its main advantage is to take into account all molec- Conclusions: RAL was well tolerated in HIV-infected patients with ESLD. There ular species (including enantiomers ans metabolites) involved in the interaction was a huge variability of raltegravir PK parameters which remained however in in vivo. the ranges reported in patients or volunteers with normal liver functions. Keywords: drug-drug interaction; cyp3a4; quantitative prediction; pharmacoki- Keywords: raltegravir, pharmacokinetics, HIV-infected patients, end-stage liver netics. disease.

12-09 12-11 Impact of CYP2D6 polymorphisms on efficacy and safety of antipsychotic A polymorphism of the aromatic L-amino acid decarboxylase gene affects drugs: a retrospective study about 52 chronic schizophrenic patients the motor response to L-Dopa in Parkinson’s disease A Boulamerya, F Richardb, JL Berge-Lefrancc, RM Richierib, C Lancßonb, D Devosa, F Cormiera, S Lejeunea, K Tahiria, F Charbonnier-Beaupela, N Rouaixa, A Enjalbertc, N Simona aService de Pharmacologie Medicale et Clinique, Facultede AM Bonneta, C Bonneta, N Zahra, J Costentina, M Vidailheta, JC Corvola aMedical Medecine, Aix-Marseille Universite, Marseille, France; bService de Psychiatrie, Hôpital Pharmacolgy, Lille, France c Ste Marguerite, Marseille, France; Plateforme de Biologie Moleculaire, Hôpital Background: In Parkinson’s disease (PD), the response to L-dopa is highly vari- Conception, Marseille, France able and unpredictable. The major pathway for dopamine synthesis from L-dopa Background: Highly polymorphic CYP2D6 are, with CYP2C19 and CYP3A, the is decarboxylation by aromatic L-amino acid decarboxylase (AAAD, encoded by major enzymes related to psychiatric drugs metabolism. CYP polymorphisms are the DDC gene). currently investigated in patients from our hospital psychiatric unit to help Objective: To determine the motor response to L-dopa in PD patients as a func- explaining antipsychotic resistance and/or side effects, and selecting right anti- tion of the rs921451 T>C polymorphism (DDCT/C). psychotic medications. Methods: Thirty-three Caucasian PD patients underwent an acute L-dopa We here report the results for evaluation of the impact of CYP2D6 poly- challenge together with the peripheral AAAD inhibitor benserazide and were morphisms on efficacy and safety of antipsychotic drugs in schizophrenic genotyped for rs921451. The primary efficacy criterion was the motor response patients. to L-dopa, as estimated by the area under the curve for the change in the United Methods: Retrospective evaluation of schizophrenic patients was performed. Clin- Parkinson’s Disease Rating Scale part III (UPDRS) score relative to baseline ical data, especially efficacy and safety, were extracted from electronic medical (AUCDUPDRS) in the 4 h following L-dopa administration. Secondary endpoints records, physician’s and patients interview. Genomic DNA was extracted after were pharmacokinetic parameters for plasma levels of L-dopa and dopamine. informed written consent in schizophrenic patients. CYP2D6 (*2, *3 *4, *6,*9, Investigators and patients were blinded to genotype data throughout the study.

Results: Fourteen patients (eight males and six females) had the DDCCC/CT geno- invasion and metastases development. Heparin has been shown to prolong sur- TT type and 19 (15 males and four females) were DDC . When adjusted for the L- vival of laboratory animals after tumor cell inoculation. Antineoplasic actions of CC/CT dopa dose, AUCDUPDRS was significantly lower in DDC patients than in heparin are mediated by anticoagulant and non-anticoagulant effects. Heparins DDCTT patients (1017.6 Æ 241.7 vs. 1369.7 Æ 131.8, respectively; P = 0.01). have significant antiangiogenic properties through the inhibition of thrombin and There were no significant intergroup differences in plasma pharmacokinetic fibrin formation. Low-molecular-weight heparins (LMWHs) also inhibit the activ- parameters for L-dopa and dopamine. ity of vascular endothelial growth factor (VEGF) and basic fibroblast growth fac- Discussion: The rs921451 polymorphism in DDC influences the central motor tor (bFGF) through the reduction of tissue factor expression, resulting in reduced response to L-dopa but does not significantly change peripheral pharmacokinetic endothelial cell growth. In most preclinical studies, LMWH did not directly affect parameters for L-dopa and dopamine. Our results suggest that DDC is a genetic primary tumor growth, but rather interfered with metastasis formation. Heparin modifier of the L-dopa response in Parkinson’s disease. intervenes in several steps of the metastatic process. Heparin reduces tumor inva- Classification of evidence: Class II. siveness by inhibiting heparinases and other extracellular matrix components. Keywords: Parkinson’s disease, dopamine, L-amino acid decarboxylase, genetic, Heparin also binds to P- and L-selectins and reduces crucial cancer cell interac- L-dopa. tions with endothelium, platelets, and leukocytes which are key components of the metastatic process. Heparin enhances tumoricidal activity of natural killer cells. 12-12 Clinical data about the potential antitumor effects of heparins are less convincing. Interindividual variability in nevirapine pharmacokinetic: impact of a Meta-analyses have suggested that, compared with unfractionated heparin, combination of CYP2B6 polymorphisms in HIV-infected Cambodians (the LMWH may reduce the mortality of patients with cancer receiving anticoagulant ANRS 12154 Study) treatment for venous thromboembolism. In a large trial comparing prolonged J Bertranda, M Choub, DM Richardsonc, PD Legerc, F Mentred, AM Taburete, LMWH treatment with a coumarin for the treatment of venous thromboembolism DW Haasf, C Verstuyfte aGenetics Institute, University College London, London, UK; in cancer patients, LMWH have been associated with an increase in overall sur- bFaculte de Pharmacie et laboratoire Rodolphe Merieux, Cambodge, Phnom Penh, vival in patients with non-metastatic cancer receiving LMWH. The meta-analysis Cambodia; cDepartement de Medecine et Pharmacologie, Centre de genetique Humaine, of the trials evaluating LMWH in patients with cancer who have no standard Universite de Vanderbilt, Nashville, TN, USA; dUMR 738, Paul Diderot Universite, indication for anticoagulant treatment shows that LMWH is associated with a Paris; eUniversite Paris Sud, Hôpital Bicêtre AP/HP, Hôpitaux Universitaires Paris relative risk of death of 0.94 (95% CI, 0.88 to 1.00). However these trials were Sud, Le Kremlin Bicetre; fDepartement de Medecine et Pharmacologie, Centre de conducted in patients with different types of cancer, most of the patients had genetique Humaine, Universite de Vanderbilt, Nashville, TN, USA locally advanced or metastatic cancer and LMWH was administered at prophy- Objective: NVP is metabolized primarily by CYP2B6 and CYP3A4. In a previous lactic dosage in most of the studies. Thus additional evaluation in specific cancer report among 4 CYP2B6 polymorphisms candidiates, we described a significant types, using higher dosages of heparin, is needed. At this time, at least six addi- association between the CYP2B6 G516T (rs3745274) polymorphism and the tional trials aiming to enroll about 3500 patients in total are ongoing and may clearance of NVP in HIV infected Cambodians who had participed in ANRS help provide the answers. 12154 study (1). In a second step, an extensive genetic study was performed for Keywords: Anti-tumor. CYP2B6, ABCB1 and NRlI2 polymorphisms. Patients and methods: Patients of this study were enrolled as a part of the Esther Cohort (Calmette hospital, Phnom Penh, Cambodia), Patients were treated 13-01 by NVP (200 mg 9 2/day) associated with lamivudine + stavudine or zidovu- Implications of using the MDRD or CKD-EPI equations instead of the dine. At 18 and 36 months, morning plasma samples for NVP assays were Cockcroft-Gault equation for drug dosage adjustment in elderly patients obtained at trough 12 Æ 2 h (114 at M18 and 124 at M36). NVP concentra- E Farniera, S Goutelleb, L Goutelle-Audibertc aHospices Civils de Lyon, Lyon, France; tions data were analyzed using a nonlinear mixed- effect model approach with bHospices Civils de Lyon & Universite Lyon 1, Lyon, France; cCentre Hospitalier de MONOLIX software version 2.4. Using this population model, empirical Bayes Saint Laurent de Chamousset, Saint Laurent de Chamousset, France estimates of individual NVP clearances (Cl/F), were derived for patients with The Cockcroft-Gault (CG) equation is the traditional method used to estimate evaluable genetic data. renal function for drug dosage adjustment. The Modification of Diet in Renal Dis- Results: Among 170 patients included in the Esther cohort treated by NVP, 129 ease (MDRD) and Chronic Kidney Disease – Epidemiology Collaboration (CKD- were included in genetic analyses (median at M36: 37 years, 56 kg, EPI) equations provide more accurate estimation of the glomerular filtration rate CD4 a 299 c/mm3, 95% with ARN-VIH plasmatic < 400 copy/mL). In the final (GFR) but they have not been validated for drug dosing. We evaluated the impli- model, 11 % of the interpatient variability in NVP CL/F was explained by these cations of using the MDRD or CKD-EPI equations in place of the CG equation for three CYP2B6 polymorphisms: CYP2B6 G516T (8.5%), rs7251950 (2.3%) and drug dosing in elderly patients. rs2279343 (0.2%). The frequencies of these polymorphisms were 0.34, 0.37 and This was a prospective study conducted in two geriatric hospitals. All inpatients 0.40 respectively. hospitalized from May 1 to August 31, 2012 with at least one serum creatinine Conclusion: Among HIV-infected Cambodians receiving NVP containing antiret- <3 days were included. Renal function was estimated using the CG, MDRD, and roviral regimens, steady state plasma NVP clearance was best described by a CKD-EPI equations. Manufacturer dosing guidelines regarding renal function combination of CYP2B6 polymorphisms. were retrieved for 36 drugs widely used in geriatric medicine. Patients’ drug Reference: treatment and dosages were examined using the CG equation as the reference 1) Chou M., Bertrand J., Segeral O., Verstuyft C., Borand L., Comets E., Le Tiec method for renal dosing adjustment. We compared 1/ the agreement between C., Becquemont L., Ouk V., Mentre F., Taburet A.-M. Low inter and intra individ- the recommended and the actual patients’ dosage regimens 2/ the estimations of ual variabilities in nevirapine pharma cokinetics in HIV-infected Cambodian renal function provided by the three equations 3/ the dosage predicted by using patients (ANRS12154). Antimicrob. Agents Chemother. (2010) 54(10) 4432– the guidelines with each equation. 4439. A total of 249 patients (158 females, 91 males, 84 Æ 7 years) were included in Keywords: HIV, CYP2B6, Nevirapine, Pharmacogenetic. the study. Mean estimates of renal function from the CG, MDRD and CKD-EPI equations were 51 Æ 24 mL/min, 72 Æ 35, and 64 Æ 22 mL/min/1.73 m2, respectively (P < 0.001). Ninety-four percent of patients had at least one drug with renal dosing recommendation. Prescribed doses were not appropriately SESSION 13: APNET: FACE CACHEE DES adjusted to renal function in 20% of patients. Compared with the CG equation- based dosages, the use of the MDRD and CKD-EPI equations for renal dosing ANTICOAGULANTS adjustment was associated with dosage discrepancy in 25% and 22% of patients and 17% and 15% of drug orders, respectively. Theses discrepancies were overdoses in 94% of drug orders. The pro-calcifying role of vitamin K antagonists in the cardiovascular Dosage adjustments to renal function are frequent in elderly patients. In such a system a a population, the use of the MDRD or CKD-EPI equation instead of the recom- G Leftheriotis PRES l’UNAM, BNMI UMR CNRS 6214-Insemr 1083, Medical mended CG equation result in dose discrepancy – mainly overdose – in 20–25% School of Angers, University of Angers, Angers, France of patients, and thus should be avoided. Arterial calcification is a recognized and independent cardiovascular risk factor. Keywords: renal function, geriatrics, drug dosage adjustment. It is commonly associated to acquire metabolic diseases such as Diabetes or chronic renal insufficiency and in rare genetic diseases. Calcification of the arterial wall is a complex and fine regulated process involving specific physico-chemical conditions, pro and anti-calcifying factors. Among these factors, vitamins play 13-02 a pivotal role. The role of vitamin D is well-known in the calcifying process of Re-evaluation of antibiotherapy before the 72 h: a regional evaluation of the peripheral conjunctive tissues, but less is known about the role of other vita- traceability in patient records P Savarya, E Remya, JM Germainb, M Lucasb, J Douceta, V Merlec aOMEDIT Haute mins, such as vitamin K. Vitamin K (including K1, K2 and K3) is a cofactor of b c enzymatic reactions (i.e. carboxylation) necessary for the central (i.e. liver) acti- Normandie, Rouen, France; ARLIN Haute Normandie, Rouen, France; Departement vation of proteins involved in the coagulation processes but also of tissue anti- d’Epidemiologie et de Sante publique, CHU – Hôpitaux de Rouen, Rouen, France calcifying factors such as matrix Gla protein. Drugs such as vitamin-K-antago- Purpose: In healthcare facilities (HF), the re-evaluation of any antibiotherapy nists (VKA) are mostly known for there anticoagulants properties, but several (ATB) before the 72 h is recommended and valued in the composite indicator of studies suggest a significant pro-calcifying role in the cardiovascular system. The good use (ICATB). The French National Authority for Health requiring its com- present topic will review and discuss this uncommon side effect of the VKA treat- prehensive traceability in the patient record . The aim was to evaluate the trace- ments. ability across all HF of one region on the occasion of the national prevalence survey (NPS) of healthcare-associated infections in 2012. Patients and methods: A questionnaire on the traceability of the re-evaluation has been proposed to all regional HF during the NPS. All patients, receiving on the day of the survey for more than 72 h a curative ATB introduced by the HF, Anti-tumor effects of low-molecular-weight heparin were included. The NPS investigators sought in the patient record the evidence G Meyera aAssistance Publique Hôpitaux de Paris, Hôpital Europeen Georges of the re-evaluation. In the absence of one official definition, explicit or implicit Pompidou, Universite Paris Descartes, Sorbonne Paris Cite, Paris, France criteria of traceability have been defined (see below) and used for all HF. Heparin was reported to interfere with several steps of tumor progression and Results: Of the 56 HF affected, 27 HF (88–2241 beds) included a total of 620 tumor metastasis, including tumor growth, tumor cell motility, migration, patients. The re-evaluation was reported in 65.2% of cases, explicitly in 50.2%

such as comments on clinical (41.0%) and / or biological (33.0%) evidence, decreases the risk of food allergy in children despite the heterogeneity of the five words ‘ATB re-evaluation’ (10.3%), a specific record sheet (4.7%). These criteria studies included. The effectiveness of SOTI to induce tolerance in children with were associated in 15.0%. The re-evaluation was only implicitly mentioned if food allergy was not associated with higher tendency of adverse events during ATB was just written without explanation or a clinical comment without the the protocol compared to allergen avoidance. consequences for ATB. Seven HF with 100% traceability had very few cases ana- Keywords: food allergy, oral tolerance induction, meta-analysis lyzed: the average of cases was 3 per HF (1–9) against 30 (2–219) for 20 other HF. A better traceability (>75%) (14 of 27 HF) was not related to the number of beds in the HF (418 beds vs. 408, P = 0.62) nor the proportion of beds with 13-05 computerized prescription (39% vs. 27%, P = 0.89), nor the ICATB grade(vs. 7A/ Evaluation of the management of major bleeding under VKA in an 14. 9A/13). emergency unit Discussion: Even with a broad definition including implicit elements, evidence of J Saint-Denisa, N Dublancheta, F Moustafaa, J Schmidta aCHU Gabriel Montpied, a re-evaluation of the ATB was only found in two cases out of three. The require- Clermont-ferrand, France ment of the French National Authority for Health to reach 100% of ATB re-eval- Introduction: The Oral anti-vitamin K anticoagulants are sill today uation seems difficult to achieve, and more if it’s considered throughout a year, the most widely used anticoagulants treatments for the prevention and treatment except for HF with very few prescriptions or full computerized system. of thromboembolic diseases. In 2008, the French National Authority for Health Keywords: antibiotherapy, re-evaluation, traceability. (1) published guidelines for the management of severe bleeding in patients on vitamin K antagonist (VKA). The aim of the study was to determine if our man- aging of bleeding events was consistent with these guidelines. 13-03 Materials and Methods: A retrospective analysis was performed on all patients Chitosan-bioactive glass improves bone microarchitecture in who received prothrombine complex concentrate (PCC) from 2010 to 2011 in osteoporosis our emergency unit. Characteristics collected were: VKA type, the International S Jebahia, O Hassanea, A Nacerb, R TAREKc, K Hassibb, EF Abdelfattahd,E Normalized Ratio (INR), location of severe bleeding, medical care and evolution Hafede aUniversite de Rennes 1, UMR CNRS 6226, Campus de Beaulieu, 263 av. du of the hemorrhage event. General Leclerc, 35042 Rennes, France; bLaboratory of Orthopaedic and Traumatology Results: During this period 264 patients received PCC for severe bleeding under medicine Faculty Sfax, Sfax, Tunisia; cLaboratory of Histology, medicine Faculty Sfax, VKA. Mean age was 78 years old [IC95 % = (76.6–79.3)]. In 24.3% patients, medicine Faculty Sfax, Sfax, Tunisia; dAnimal Ecophysiology Laboratory, Sciences INR was beyond the upper value of the target range. The two main locations for Faculty of Sfax, Department of Life Sciences Sfax, sfax, Tunisia; eLaboratory of Science bleeding events were intracranial hemorrhage (25.1%) and gastrointestinal hem- Materials and Environnement Faculty Sfax, sfax, Tunisia orrhage (33%). Intracranial hemorrhage diagnostic confirmation needed longer Background: Bioglass (BG) has been used to repair bone defects. Chitosan (CH) time without statistical significance (3 h25 vs. 4 h21 for other bleeding loca- administration is proposed to prevent the decrease in bone mineral density (BMD) tions, P = 0.166). One hundred and ninety five patients (81.2%) were given Vita- by inhibiting osteoclastic cells and preventing bone loss [1]. The association min K and 169 patients (70.4%) received the recommended dose of PCC with, between chitosan and glass has been found to rebalancing bone turnover in favor respectively, median times for administration of 15 min and 55 min. After diag- of formation [2]. nosis, Vitamin K and PPC were given after respectively 47 min [(34–60)] and Objectives: The present study aims at investigating the effects of the association 79 min [(68–90)]. The dose of vitamin K and PCC followed the guidelines for between chitosan and bioactive glass in bone architectural and healing processes. 110 (46.1%) patients. But the mean time between admission and PPC adminis- Methods: Female Wistar rats were ovariectomised and implanted with BG (OVX tration was 3 h57 [3:29–4:25]. The hospitalization rate was 97.7%, with a mean -BG), BG-CH (OVX -BG-CH) . A drilled hole was created on the lateral aspect of duration of 15.1 days (IC95% = 12.5–17.8). After seven days, mortality rate was the femoral condyle. After 60 days of implant insertion, all rats were sacrificed 12% and 169 patients were still hospitalized. and specimens were harvested for biological evaluation. BV/TV, N.Ob and Ob.S/ Conclusion: The treatment of severe bleeding under VKA in our unit is satisfac- BS, were measured by a point count method [25] using a 25-point integrating fil- tory according to the french guidelines. Furthermore, efficiency has been ter. improved since our 2009 study based on similar population and parameters. Results: The quantitative analysis demonstrated that Bone/Tissue Volume BV/ Severe bleeding under VKA is correlated to a high in-hospital mortality and TV, Osteoblast Number N.Ob and Osteoblast/Bone Surface Ob.S/BS were changed needs a standardized and early treatment. respectively by 5, 25.2 and 19 % in BG-CH groups when compared to those of Keywords: VKA, hemorrhage, Prothrombine complex concentrate BG group. Moreover, the level of the basic structural unit wall width and the trabecular hypertrophy showed a pronounced increase. In fact, BG-CH was shown to limit bone loss inducing a beneficial impact on the trabecular bone microarchi- tectural properties. It was confirmed by the reduced inter-trabecular space and SESSION 14: A3P: COMMUNICATIONS LIBRES the higher trabecular thichness when compared to those of BG-treated rats. Conclusion: The incorporation of 17%W of CH in BG respects the physiological bone formation and ameliorates the bone architecture and the healing processes. 14-01 This study has added that re-build cancellous structure with BG-CH implant is a Survival after hemorrhagic stroke following oral antithrombotic agents promising candidate repair/antioxidation/ regeneration therapy. therapy: a retrospective study References: A Gourauda, G Clavea, N Voirinb, D Rerbalc, BA Kimd, B Valleee, C Payena,T 1. Ezoddini-Ardakani F., Azam A., Yassaei S., Fatehi F., Rouhi G. Effects of chito- a a – Vial Poison Center and Pharmacovigilance Department, Hospices Civils de Lyon, san on dental bone repair (2011) 3, 200 205. Lyon, France; bCNRS UMR 5558, Epidemiology and Public Health Group,Lyon 2. Jebahi S., Oudadesse H., Bui X., Keskes H., Rebai T., el Feki A., el Feki H. University Hospitals, Lyon, France; cMedical Emergency Unit, Edouard Herriot Repair of bone defect using bioglass-chitosan as a pharmaceutical drug: An Hospital, Hospices Civils de Lyon, Lyon, France; dGeriatric Department, Edouard experimental study in an ovariectomised rat model. (2012) 161276, 1287. Herriot Hospital, Hospices Civils de Lyon, Lyon, France; eDepartment of Neurosurgery Keywords: chitosan-associated bioactive glass, Ovariectomised rats, bone healing B, Pierre Wertheimer Hospital, Hospices Civils de Lyon, Lyon, France Objective: Oral antithrombotic agents are associated with an increased risk of hemorrhagic events among which those arising in the central nervous system 13-04 are the most severe. However, the exact contribution of oral antithrombotic treat- Specific oral immunotherapy for food allergy in children: a systematic ment to a fatal outcome after intracranial hemorrhage remains controversial. review and meta-analysis of randomized control trials The aim of this study was to investigate whether exposure to an oral antithrom- C Lamottea, C Predab, C Iliescuc aClinical Investigation Centre, CIC 9301 CH&U- botic drug at the time of cerebral hemorrhage can influence early survival. Inserm; Department of Internal Medicine; Centre Hospitalier Regional Universitaire, Methods: A 1-year monocentric retrospective comparative study was conducted Lille, France; bDepartment of Applied Mathematics, UMR CNRS 8524, Lille 1 in Lyon university hospitals. Cases of intracranial hemorrhage were identified University, Lille, France; cPediatrics Department, Allergology Centre, Lille Catholic through the hospital discharge database by using an algorithm derived from the Hospitals, Lille, France 10th version of the International Classification of Diseases (ICD 10). Exposures to Aim: To assess the effectiveness and safety of specific oral tolerance immunother- oral antithrombotic agents were identified after manual checking of all hospital apy (SOTI) compared to allergen avoidance in children with food allergy. charts. Survival at days 7 and 30 was compared according to the type of pre- Design: Systematic review and meta-analysis. injury exposure (vitamin K antagonist or, antiplatelet treatment, or no anti- Data sources: Medline (1992 to October 2012), reference list of retrieved arti- thrombotic treatment) using the Kaplan-Meier method. A multivariate Cox model cles. was performed to assess predictive factors of mortality at day 7. Study selection: Randomized controlled trials (RCT) of SOTI vs. allergen avoid- Results: Two hundred and one patients with confirmed hemorrhagic stroke were ance with the following criteria: 1) Clinical history of Immunoglobulin E medi- retained for analysis (51 treated by antiplatelet, 59 by vitamin K antagonist and ated food allergy diagnosed using double blind placebo-control food challenge, 2) 91 without antithrombotic treatment). Cardiac failure, a history of previous Three phases SOTI protocol (induction, build-up, maintenance), 3) Children aged ischemic stroke and intracerebral hemorrhage with intraventricular spread or 0–18 years, 4) Final status (tolerance/allergy) and safety data reported in multifocal intracranial hemorrhage were significantly more frequent in the vita- patients and control groups. Two authors independently extracted the data. min K antagonist and the antiplatelet groups as compared to unexposed patients Results: Five RCTs (240 children) met our inclusion criteria. Four studies (68.4% and 30 %, respectively, vs. 11.1%, 17.9% and 28 % vs. 5.6 %, 35.6% showed a statistically significant reduction in endpoint allergy after SOTI vs. con- and 29.4 % vs. 16.5%). 30-day survival was significantly lower in vitamin K trol. All pooled analyses were based on random-effects models. A significant het- antagonist exposed patients compared to unexposed patients. By contrast, anti- erogeneity between the studies was observed (Q = 31.79, P < 0.001; I2 = 87.4%, platelet agents were not associated with a significant increase in early mortality. 95% confidence interval (CI), 73–94.1%). The relative risk (RR) of allergy was Multivariate Cox model suggested that a previous history of ischemic stroke (Haz- lower after SOTI (RR = 0.36; 95% CI, 0.18–0.75). Adverse events (AE) were ard Ratio=3.03; 95%CI 1.06–8.71) and the initial severity of bleeding, i.e. intra- reported by 4 studies. Compared with controls, the RR of AE during SOTI ranged ventricular spread or multifocal location (HR = 8.99 95%CI 1.95–41.49) were from to 0.18 to 2.33 and no significant correlation with RR of allergy was the main predictors of mortality at day 7 in patients treated by vitamin K antago- detected. AE were considered mild or moderate in 221/240 patients and epineph- nists. rine was administrated in 7/240 patients; only one patient (control group) had Conclusion: Vitamin K antagonists, but not anti-platelet agents, increased the anaphylactic shock by accidental ingestion. risk of death from hemorrhagic stroke during the first 30 days after hospitaliza- Discussion and conclusions: Our meta-analysis, considering two additional tion. RCTs with respect to the Fisher’s one (2010), supported the evidence that SOTI Keywords: hemorrhagic stroke, early survival, antithrombotics

14-02 reported [2]. We report a series of four patients with proven pholcodine-NMBA Evaluation of knowledge and use of prescription tools by hospital cosensitization. physicians at the teaching hospital of Bordeaux Observations: The first case was a patient who experienced NMBA anaphylaxis A Fauchera, D Berda€ıa, C Brotonsa, A Fourrier-Reglata, A Parientea, and whose history revealed that 2 months before the surgery she had experienced N Moorea aUniversite Victor Segalen Bordeaux 2, Bordeaux, France generalized urticaria with palpebral edema 2 h after the ingestion of pholcodine Objective: To evaluate knowledge and use of prescription tools by hospital physi- syrup. Skin tests (prick tests and intradermal tests) confirmed pholcodine and cian at the teaching hospital of Bordeaux. suxamethonium allergy, with cross-reactivity to rocuronium and vecuronium. Methods: A cross-sectional study was conducted between March and August The other cases concerned three patients referred to the hospital following phol- 2012 among 240 randomly selected physicians of the teaching hospital of Bor- codine anaphylaxis. Allergy tests confirmed pholcodine allergy and revealed deaux. The sample was composed of 60 interns, 60 residents, 90 consultants NMBA sensitization despite no history of NMBA anaphylaxis. Moreover, in 2 of (PH), 10 associate professors and 20 full professors. Each selected physician has them, basophil activation test was positive for suxamethonium. been asked to fill in a standardized questionnaire exploring their knowledge and All NMBA skin prick tests and intradermal tests were performed according to usage of prescriptions tools available in the hospital (i.e. the French drug formu- published recommendations [3]. lary, recommendations from French health authorities, local recommendations Discussion: These cases of pholcodine and NMBA cosensitization provide evi- elaborated by the hospital drug committee, etc.). Data were collected through dence of sensitization via pholcodine exposure. The skin reactivity towards individual face-to-face interviews. NMBAs and the positive basophil activation test for suxamethonium in the 2 Results: A total of 90 out of 240 physicians participated in the survey: 29 patients referred for pholcodine allergy suggest that the presence of specific IgE to interns, 23 residents, 26 consultants, two associate professors and 10 full profes- pholcodine can induce mastocyte degranulation in the presence of NMBA. sors. All respondents reported to use the French drug formulary. Good practice Considering these results, NMBA should be skin tested before anesthesia in recommendations for drug use (from by health authorities or learned societies) patients with confirmed pholcodine allergy. Moreover, any suspicion of pholco- were reported by all physicians to be used in their practice with the exception of dine allergy should be confirmed by skin tests. interns (55%). The Bordeaux University Hospital’s drug formulary (limited list) References: was known by 70% (N = 63) and used by 14% (N = 13) of physicians. The hos- 1. Florvaag E., et al. Prevalence of IgE antibodies to morphine. Relation to the pital drug committee was known by 41% of physicians (N = 37). 80% of physi- high and low incidences of NMBA anaphylaxis in Norway and Sweden, respec- cians (N = 72) reported prescribing off-label. 74% of physicians (N = 67) felt that tively. Acta Anesthesiol Scand (2005) 49 437–444. their initial training did not prepare them appropriately for the act of prescribing. 2. Codreanu F., et al. Allergy to pholcodine: first case documented by oral chal- Discussion: The results of this study show that the knowledge and use of pre- lenge. Allergy (2005) 60 544–545. scription tools of the Bordeaux University Hospital can be improved. Several fac- 3. Mouton-Faivre C., et al. Realisation pratique du bilan allergologique cutane a tors may explain these results: the difficulty of access to these tools, their lack of visee anesthesique, dans le respect des recommandations pour la pratique cli- adaptation to clinical practice and lack of communication. Simplification, nique: qui tester, quoi tester, comment tester? Rev Fr Allergol Immunol (2003) improved accessibility and updating of prescription tools should allow better 43 281–288. appropriation by physicians. Keywords: pholcodine, neuromuscular blocking agents, anaphylaxis Keywords: Prescription, drugs, hospital, hospital physician, good practice recommendations 14-05 Quetiapine-induced ischemic colitis: review of the French 14-03 Phamacovigilance database Drug interactions between pristinamycin and antivitamine K: 22 cases V Pinzania, H Peyrierea, L Javotb, S Horowiczc, T Bejan-Angoulvantd,MN collected by the french pharmacovigilance centres Network Osmonte, M Tebacher-Altf, H Le Louetg, C Le Bellerh, D Hillaire-Buysa aCRPV AL Guibouxa, S Combretb, A Spreuxc, A Gouraudd, T Trenquee, M Tebacher-Altf, Languedoc-Roussillon, Montpellier, France; bCRPV Nancy, Nancy, France; cCRPV C Sgrob aCRPV, Dijon Cedex, France; bCRPV, Dijon, France; cCRPV, Nice, France; Fernand Widal, Paris, France; dCRPV Tours, Tours, France; eCRPV Rennes, Rennes, dCRPV, Lyon, France; eCRPV, Reims, France; fCRPV, Strasbourg, France France; fCRPV Strasbourg, Strasbourg, France; gCRPV Henri Mondor, Paris, France; Pristinamycin is a streptogamin antibiotic authorized in France since 1973 and hCRPV HEGP, Paris, France active especially on Staphylococcus aureus and Streptoccus pneumoniae. Its indi- Introduction: Xeroquelâ (quetiapine) is a recent antipsychotic drug marketed in cations were restricted in July 2012 to acute maxillary sinusitis, acute exacerba- France since November 2011. Shortly after the availability of quetiapine, the first tion of chronic bronchitis, mild or moderate community pneumonias, ischemic colitis was reported to the Pharmacovigilance centers. Most antipsychot- dermatological and smooth tissue infections. Drug interactions with oral antico- ics commonly cause gastrointestinal hypomotility and intestinal necrosis may be agulant currently described in the Summary of Product Characteristics mention a severe consequence of such gastrointestinal disturbances. ‘increase of anticoagulant activity has been reported with antibiotics in general, Methods: We reviewed all observations of quetiapine-induced ischemic colitis or and more particularly with others classes of antibiotics than pristinamycin’. To gastrointestinal necrosis, notified to the French Pharmacovigilance centers. Then date, our research failed to find any case report of interaction between pristina- we enlarged our research to other antipsychotics and finally we compared the mycin and antivitamin K drugs (AVK). data of the previous review published by Peyriere et al. in 2009 [1]. We report here 22 cases of increase AVK effects in patients treated with pristina- Results: Eight cases of ischemic colitis and gastrointestinal necrosis associated mycin collected by the French Pharmacovigilance Centres between 1999 and with quetiapine were analysed (four men and four women). The mean age was 2012. 34 years [20–66]. In four cases, digestive complications led to partial or total Material and method: All cases including pristinamycin and one AVK (fluindi- resection of the colon and two patients died despite surgery. Among risk factors, one, warfarin or acenocoumarol), and international normalized ratio (INR) in 6/8 (75 %) cases quetiapine was associated with antipsychotics and also with increase, and/ or prothrombin time (PT) decreased, and/or hemorragic symptoms atropinic-like drugs. The mean delay of quetiapine-induced ischemic colitis was were extracted from the French Pharmacovigilance Database. Cases containing 30.8 days [10–81]. another suspected interacting drug with AVK or known for causing hemorrhage In the French database, 38 cases are reported until the end of 2006 and it has (such as heparins) were excluded. been pointed out that 55% of patient received more than one antipsychotic and Results: Twenty-two cases were included (nine men and 13 women, mean age 68% of patients had an association with other atropinic-like drugs [1]. Since this 75.8 years (36–89). All patients were previously treated with fluindione (77.3%), time period, 27 additional cases are described (including quetiapine cases), 81.5% acenocoumarol (18.2%) or warfarin (4.5%) before pristinamycin introduction. were associated with antipsychotics and 48.2% received an association with The mean time between pristinamycin onset and adverse effect was 7.7 days (2– other atropinic-like drugs. 18 days). Adverse effects were increased INR or decreased PT without haemor- Conclusion: Quetiapine, a recent antipsychotic drug that is presented with low rhages (11 cases / 50%), with haemorrhages (10 cases/45.5%), 1 haemorrhage atropinic-like activity is also involved in gastro-intestinal hypomotility. Intestinal without INR or PT value. Fifteen cases were serious. Evolution was favourable necrosis associated with antipsychotics is very rare; however mortality is high, (86%) or unknown (14%). with a rapid worsening of patients towards septic shock. It is therefore essential Discussion: These cases strongly suggest that pristinamycin increases AVK to add this adverse-effect to medical recommendations to draw attention of physi- effect. Mechanisms of this interaction are unknown. Intra erythrocyte enzymes cians about these complications and the associated signs. destroy Pristinamycin. It is probably metabolized by the liver (but the pathway is Reference: still unknown), and seems also able to link and inhibit P-Glycoprotein. However 1. Peyriere H., Roux C., Ferard C., Deleau N., Kreft-Jais C., Hillaire-Buys D., Bou- we failed to find any link between AVK and P-Glycoprotein. lenger J.P., Blayac J.P.; French Network of the Pharmacovigilance Centers. Anti- Conclusion: Whatever the mechanism, our study exhibits the existence and the psychotics-induced ischaemic colitis and gastrointestinal necrosis: a review of the severity of this poorly known drug interaction. This should be specified in the French pharmacovigilance database. Pharmacoepidemiol Drug Saf. (2009) 18 Summary of Product Characteristics with recommendations of clinical and INR 948–955. increased monitoring. Keywords: quetiapine, antipsychotics, ischemic colitis Keywords: pristinamycin, Antivitamin K, interactions

14-06 14-04 Metronidazole-induced peripheral neuropathy: a case series Pholcodine and neuromuscular blocking agents cosensitization N Pareta, A Grandvuilleminb, MJ Jean-Pastorc, F Beau-Salinasd, C Guye, N Petitpaina, JM Renaudinb, E Beaudouinb, L Javotc, D Swiegotd, M Beckerd, J Descotesa, T Viala aCentre Regional de Pharmacovigilance - Centre anti-poison - J Scala-Bertolac, P Gilletc, PM Mertese aPharmacovigilance Regional Center of Hospices Civils de Lyon, Lyon, France; bCentre Regional de Pharmacovigilance - CHU Lorraine, Nancy cedex, France; bDepartment of Allergology, Emile Durkheim Hospital, de Dijon, Dijon, France; cCentre Regional de Pharmacovigilance - Assistance Publique - Epinal, France; cDepartment of Clinical Pharmacology and Toxicology, CHU Nancy, H^opitaux de Marseille, Marseille, France; dCentre Regional de Pharmacovigilance - Nancy, France; dPharmacovigilance Regional Center of Lorraine, Nancy, France; CHU de Tours, Tours, France; eCentre Regional de Pharmacovigilance - CHU de Saint- eDepartment of Anesthesiology, CHRU Strasboug, Strasbourg, France Etienne, Saint Etienne, QC, Canada Introduction: The use of neuromuscular blocking agents (NMBAs) can cause Objectives: Long-term or high-dose metronidazole treatment is associated with anaphylaxis even the ﬁrst time they are used in a given patient. Possible sensiti- peripheral neuropathy and the potential persistence of symptoms for a long per- zation to NMBAs resulting from pholcodine exposure has been proposed in 2005 iod of time. A recent case of disabling peripheral neuropathy that persisted over (pholcodine hypothesis) [1], although allergic reactions to pholcodine are rarely 4 years in a patient who continued metronidazole more than 1 year after the

onset of symptoms prompted us to review and characterize spontaneous reports Patterns of use were compared with the SmPC and guidelines (French National of peripheral neuropathy with particular attention to possible risk factors. Gastro-Enterology Society, SNFGE, 2009). Material and methods: Cases suggestive of peripheral polyneuropathy with Results: A total of 530 patients were included in the cohort (301 in 2009, 229 metronidazole as a suspected drug and reported between 2002 and 2012 were in 2010). Investigation of KRAS status was performed in 95.0% of patients extracted from the French pharmacovigilance database. Reports with insufficient included in 2009, and in 99.6% of patients in 2010. KRAS investigation was information, onset of symptoms more that 1-month after the end of metronida- performed on primary tumour (65.4%), metastases (12.1%), or both (1.0%). Fre- zole treatment, underlying cancer, and short-occurring or transient paresthesias quency of wt-KRAS gene in patients treated with CTX was similar in 2009 were excluded. (96.5%) and in 2010 (96.9%). A total of 389 wt-KRAS patients were followed- Results: Thirty-seven cases (23 women and 14 men; mean age: up: 76.9% were treated every 2 weeks, 56.0% were treated with irinotecan-based 52.2 Æ 19 years) were retained. The daily dose was greater than or equal to the regimen, 37.8% with oxaliplatin-based regimen, and 6.2% with another regimen. maximal recommended dose (1.5 g/day) in 25 of 29 (86%) patients. Symptoms As compared with the guidelines, 15 patients (0.03%) with mutated KRAS occurred with a median delay of 60 days (range: 3–584) after starting metroni- received; four of these received an oxaliplatin-based regimen considered as delete- dazole and the median cumulative dose was 90 g (range: 4.5–584, n = 27). rious to survival outcome by the SNFGE. In addition, 98.5% of all patients were Peripheral neuropathy was confirmed by electromyogram in 16 patients. Cerebel- treated without prior investigation of EGFR expression, and one patient was trea- lar symptoms or balance disorders were concomitantly noted in 8. In 35 patients, ted by Folfox-bevacizumab associated with CTX. metronidazole was discontinued within a median delay of 11 days after the onset Discussion: EREBUS is the first post-marketing cohort conducted in France to of symptoms, and 1 only had dose reduction (data unspecified in 1). A 1-year fol- describe the usage patterns of CTX. There was a high level of conformity to the low-up was available in 14 patients. Ten fully recovered or dramatically SmPC and to the SNFGE guidelines regarding KRAS status and modalities of use. improved whereas four still had persistent disabling or worsening symptoms. As Investigation of EGFR expression was infrequently performed which is in confor- compared to those who recovered, these last patients more frequently continued mity with the SNFGE guidelines that no longer recommend immunohistochemis- metronidazole for more than 15 days after the onset of symptoms (75% vs. 25%) try test owing to the reported poor predictive value of this parameter for and received higher mean cumulative metronidazole dose (379 g vs. 171 g, treatment response. respectively, P = 0.2). In two patients, signs of peripheral neuropathy recurred or Keywords: colorectal cancer, cetuximab, pharmacoepidemiology worsened after metronidazole reintroduction. Discussion: Although our sample is limited, our results confirm that the persistence or worsening of peripheral neuropathy may be related to high cumulative 14-09 dose and/or delayed discontinuation of metronidazole after the onset of symp- Food supplements bought on the Internet: be careful to sibutramine- toms. induced acute psychosis! Keywords: Metronidazole, Peripheral neuropathy D Faurea, N Selvya, C Philibertb, V Bresa, D Hillaire-Buysa, O Mathieua aCHRU Montpellier - Universite Montpellier 1, Montpellier, France; bCHRU Montpellier, Montpellier, France 14-07 Introduction: Since the 2000s, the purchase of food supplements on the net is Hypersomnia associated with isotretinoin: is this association relevant? growing. We report here the case of a 40 year old woman who ordered two her- JL Failliea, V Pinzania, J Jacquemoirea, S Boixa, M Tebacher-Altb, bal products for slimming and who presented an acute psychotic state. D Hillaire-Buysa aCRPV Languedoc-Roussillon, Montpellier, France; bCRPV Case report: The patient was hospitalized for acute psychotic reaction after Strasbourg, Strasbourg, France ingesting two slimming herbal products during 3 months: Irem Bitkisel Zayiflama Introduction: Oral isotretinoin is an effective treatment for severe acne. Mecha- and New Nordic Zuccarin Murier. She presented a paranoid psychotic decompen- nism involves activation of nuclear retinoid receptors that regulates cell prolifera- sation associated with cleaning obsessive-compulsive disorders. In the same time, tion and differentiation. Neuropsychological adverse reactions such as depression, she lost 16 kg (11 kg during the last month). She showed hypokaliema associ- suicidal ideation and psychosis have been described. Recent literature suggests ated with an inflammatory syndrome. Care support was symptomatic antipsy- that sleep disorders may be also associated (1–3). Herein, we report a case of hy- chotic and sedative treatment. She was released after checking her constants one persomnia associated with isotretinoin and a review of the French PharmacoVigi- week after the hospitalization. Two months later, she rescued and neuroleptics lance database. were stopped. Case report: A 15-year-old female patient was treated with oral isotretinoin Screening analysis performed in the Toxicology Unit of Lapeyronie Hospital did 40 mg/day for acne vulgaris for 17 months between September 1998 and May not find any compounds used for weight loss. Further analysis were then 2002. Since September 2001, she reported disabling hypersomnia (up to 13 h of required to the Agence Nationale de Securite des Medicaments et des produits de sleep/24 h) with diurnal fatigue. Symptoms persisted after discontinuation of the sante (ANSM)1. Analysis of Irem Bitkisel Zayiflama showed the presence of two treatment and contributed to decreased academic and professional performance. pharmacologically active compounds : sibutramine and phenolphtalein. These Patient also reported chronic diarrhea. Since 2006, she was treated with venla- two products are compounds listed in repertories I and II of the poisonous sub- faxine but symptoms persisted. In March 2011, polysomnography and actimetry stances. Sibutramine is the active ingredient of Sibutralâ, a drug withdrawn from were performed but did not confirmed hypersomnia. After 11 years, her condi- the market in January 2010 due to adverse effects such as cardiovascular events. tion improved and she totally recovered in September 2012. Phenolphtalein is a prohibited substance in medicine in France since 1999 Discussion: In 2012, three others cases of hypersomnia with isotretinoin have because of its potential carcinogenic effect. Sibutramine is an appetite suppressant been reported in the French PharmacoVigilance database. For two cases, symp- structurally related to amphetamine which has been associated with psychotic toms are ongoing 10 months and 4 years after treatment discontinuation. In one symptoms, including hyperactivity, psychomotor agitation, impulse activity and case, depression, eat disorder and hallucinations were associated. In a case paranoia. report, Kleine-Levin Syndrome, a rare condition characterized by recurrent hyper- Discussion: Medicinal products purchased on the net have the great disadvan- somnias associated with hyperphagia, has been related to isotretinoin treatment tage of coming from sources whose safety has not been proven. The vigilance of (2). Median duration of this syndrome was 8 years (4). Further studies are national and supranational agencies must be particularly firm and stringent in needed to investigate isotretino€ın-induced hypersomnia. comfort products such as weight loss products and sex stimulants. References: Reference: 1. Shehi G.M., Bryson W.J. Hypersomnia associated with isotretinoin in a patient 1. Rebiere H., et al. Detection of hazardous weight-loss substances in adulterated with recurrent major depressive disorder and acne vulgaris. Sleep. (2004) 27 slimming formulations using ultra-high-pressure liquid chromatography with 821. diode-array detection. Food Additives and Contaminants (2012) 29 161–171. 2. Smedje H., Schwan S., Hallberg E., Hallberg P. Onset of Kleine-Levin Syndrome Keywords: Sibutramine, phenolphtalein, herbal products, psychosis in association with isotretinoin treatment. Acta Paediatr. (2010) 99 946–948. 3. Ismailogullari S., Ferahbas A., Aksu M., Baydemir R., Utas S. Effects of isotretinoin treatment on sleep in patients with severe acne: a pilot study. J Eur Acad 14-10 Dermatol Venereol. (2012) 26 778–781. An emergency case of acute voluntary poisoning with trimebutine 4. Arnulf I., Zeitzer J.M., File J., Farber N., Mignot E. Kleine-Levin syndrome: a (Debridatâ) systematic review of 186 cases in the literature. Brain (2005) 128(Pt 12) 2763– S Paina, F Chavanta, B Fauconneaua, B Brunetb, JY Lardeurc, P Murab, 2776. MC Perault-Pochata aCEIP-A, Service de Pharmacologie clinique et Vigilances, CHU, Keywords: isotretinoin, hypersomnia, adverse effect Poitiers, France; bService de Toxicologie et Pharmacocinetique, CHU, Poitiers, France; cService Urgences-SAMU-SMUR, CHU, Poitiers, France Background: Voluntary intoxication with the antispasmodic trimebutine (Debri- 14-08 datâ) seems uncommon and toxic effects of this drug are not well documented in Real-life use of cetuximab as 1st-line treatment of metastatic colorectal the literature. cancer in France: evaluation of the conformity with SmPC and Methods: This case was obtained from PMSI (Programme de Medicalisation des guidelines – results of the EREBUS cohort study Systemes d’Information) codes on emergency database ‘Resurgences’. M Rouyera, A Fourrier-Reglatb, E Mitryc, E Francßoisd, E Bignona, A LE Monies de Case report: We report here a case of acute voluntary poisoning with trimebu- Sagazana, J Jovea, N Mooreb, P Noizee, D Smithe aUniversite Bordeaux Segalen - tine. CICP 0005, Bordeaux, France; bUniversite Bordeaux Segalen - CHU de Bordeaux - A 57-year old man, suffering from hepatitis C and treated by propranolol LP CICP 0005, Bordeaux, France; cInstitut Curie, Saint Cloud, France; dCentre 160 mg for cardiac troubles, was admitted to the emergency department of Poi- Lacassagne, Nice, France; eCHU de Bordeaux - CICP 0005, Bordeaux, France tiers Hospital in September 2012 during the afternoon. He has taken about 30 Background: Cetuximab (CTX) has demonstrated improved survival outcomes pellets of trimebutine in a context of familial conflict. in metastatic colorectal cancer (mCRC). CTX was first launched in 2004 as 2nd- On arrival at the emergency department, the patient became drowsy but arous- line therapy in mCRC with EGFR-expression. In July 2008, this indication was able after stimulation. Quickly, myosis appeared with breathing pauses and extended to 1st-line therapy and restricted to mCRC patients with wild-type (wt) sweating. The patient had difficulties to stay awake. KRAS gene. Here, the patterns of CTX use in a real-life setting and conformity Classical toxicologic analysis was negative (alcohol, benzodiazepines, barbiturates, with guidelines were investigated using the EREBUS cohort. acetaminophen, salicylates…) and the trimebutine assay confirmed an important Methods: EREBUS is a French multicentre (n = 92) cohort. Patients initiating intake. CTX 1st-line therapy for unresectable mCRC in 2009 or 2010 were identified After injection of a loading dose of naloxone (Narcanâ), the patient recovered a from hospital pharmacy dispensations and followed 12 months to evaluate the normal state of consciousness. Biochemical results showed moderate hepatic rate of metastases resection, usage patterns, safety and effectiveness of CTX. cytolysis (probably due to hepatitis C) and thrombocytopenia.

The patient received continuous doses of naloxone during 24 h and lactulose 16-02 (Duphalacâ) to prevent hepatic encephalopathy. The issue was favorable. N-acetylcystein triggers a mitochondrial hormesis mechanism improving Conclusion: Except a serious case published in 2011, toxicological effects of cellular antioxidant capacities trimebutine seem to be unknown. It’s necessary to draw attention of the medical F Singha, AL Charlesa, AI Schlagowskia, D Fumagallia, F Piquarda, B Genya, community on such medicines rarely used for autolysis and usually considered as J Zolla aUniversite de Strasbourg - EA3072-Mitochonchondrie, stress oxydant et safe drugs. protection musculaire, Strasbourg, France Keywords: antispasmodic intoxication, autolysis, reanimation Oxygen is important for survival, but is also crucial for redox-mediated adaptations. Central to metabolic flexibility of skeletal muscle is the mitochondrion. Mitochondrial hormesis is a phenomenon that enables the activation of mitochondrial biogenesis and thus improves the cellular and mitochondrial antioxi- SESSION 16 : GS NUTRITION, ALIMENTATION, dant capacities in response to a moderate pharmacologically induced oxidative stress. The goal of our study was to show that N-Acetylcystein (NAC), a precur- GUSTATION sor molecule of the glutathione, is able to induce mitochondrial hormesis. We have carried on some in vitro experiments on L6Woody myoblasts cell cultures, and studied the effects of NAC after different exposition times. NAC inhibited the Role of lipids in the central control of energy homeostasis maximal mitochondrial respiration (À19% at 0.5 mM, P < 0.05; À28% at 1 mM, C Cruciani-Guglielmaccia aUnite “Biologie Fonctionnelle & Adaptative” (BFA), P < 0.01 and À53% at 5 mM, P < 0.001). After 24 h of incubation with NAC Equipe HERGE, 4 rue Marie Andree Lagroua Weill Halle, Paris, France 1mM, there was an increase of the reactive oxygen species (ROS) production Lipids have been considered for decades as energy storage molecule; however it measured by EPR (Electron Paramagnetic Resonance; +42%, P < 0.001). The has been established that they also have a signaling function. In brain, fatty oxidative stress seems to trigger the activation of the mitochondrial biogenesis acids are able to modulate neuronal activity in areas involved in the control of pathways after 24 h of incubation, as shown by RT-PCR (mRNAs of PGC-1b: food intake and the regulation of glucose homeostasis, such as hypothalamus: +215%, P < 0.001; NRF1: +153%, P < 0.01; TFAm: +158%, P < 0.001). After this phenomenon is called ‘lipid sensing’. We will review the physiological role of one week of exposition, we found a decrease of ROS production compared to con- fatty acid in brain, as well as pathophysiological aspects that may occur during trol cells (À38% of ROS production; P < 0.001), an increase of mitochondrial obesity, through in vitro and in vivo studies using animal models. A special focus content (mtDNA: +144%; P < 0.05) and a significant increase of maximal mito- will be made on neuronal proteins that mediate the lipid sensing: membrane pro- chondrial respiration (+17%; P = 0.05). The activation of mitochondrial biogene- teins (GPCR, FATP, FAT/CD36) and intracellular ones (ACS, CPT1, NOS, PPARs). sis was maintained after 7 days of treatment (mRNA of PGC-1b: +122%, In addition, the action of lipoprotein lipase (LPL) in brain could be key step in P < 0.05; NRF1: +121%, P < 0.05; NRF2a: +62%, P < 0.05; NRF2b: +106%, lipid sensing as it provides fatty acids to neuron; its central inhibition has been P < 0.01; and TFAm: +179%, P < 0.01). In conclusion, NAC seems to trigger shown to provoke obesity through metabolic changes. A focus will be made on the activation of the mitochondrial biogenesis mechanism through a mitochon- extra-hypothalamic areas that are involved in the control of energy homeostasis drial hormesis mechanism. Activation of this phenomenon represents an interest- and may be a target of lipids : the hippocampus has been shown to control food ing research field in order to improve both cellular metabolism and muscular intake and body weight regulation, it is the main brain site for the expression antioxidant defenses. and activity of LPL. Keywords: Mitochondrial biogenesis, N-Acetylcystein, Hormesis, Reactive Oxy- Keywords: Lipids gen Species

16-03 Brain imaging during feeding behaviour Assessment of the prevalence and the metabolic profile of metabolic a a obesity and phenotypic obesity CH Malbert Unite Alimentation & Adaptations Digestives, Nerveuses et a a a Comportementales, Saint-Gilles, QC, Canada I Merahi , A Messaoudi EHU 1er Novembre 1954, Oran, Algerie Within the last decade, fMRI data in humans had point out the critical role of Background: With the emergence of the concept of metabolically obese normal the hedonic component in the control of food intake. Taken into account the weight vs. metabolically healthy obese individuals, the relationship between obes- massive knowledge existing on the importance of the hypothalamus on virtually ity and metabolic disorders has been a subject of some dispute. the same target i.e. weight control, it has been postulated that food intake was We estimated the prevalence of metabolically obese nonobese (MONO), metaboli- under the control of two independent networks: the hedonic and the homeostatic cally healthy obese (MHO), metabolically obese obese (MOO), and metabolically networks. Recent data obtained using functional imaging in humans and in ani- healthy nonobese (MHNO) subjects in an apparently healthy adult population, mal’s models suggests that the situation is more complex. First non painful gas- and compared the metabolic profile of these subtypes of obesity. tric distension mimicking that occurring immediately after the meal is not only Methods: Study subjects n = 64 underwent anthropometric measurements (BMI, able to activate nuclei directly associated to ascendant projections from the dorsal waist and hip circumference) and biochemical tests (glucose, triglycerides, total vagal complex but also limbic areas including prefrontal and orbito-frontal cortic- cholesterol, HDL cholesterol, uric acid, insulin and C-peptide). A lipoprotein elec- es. Second, we recently demonstrated that both duodenal and portal glucose infu- trophoresis was performed to assess the lipid profile et detect the presence of Lp (a). sions activated the dorsolateral prefrontal cortex and the primary somatosensory 2 cortex. Third, Wang team has demonstrated in some obese individuals a Obesity was defined as a body mass index (BMI) 30 kg/m , based on the WHO decreased activity of the prefrontal cortex. We have showed using a large animal guidelines. The metabolic obesity was diagnosed whenever criteria for metabolic model, that this decreased activity is an acquire feature of obesity and not a pre- syndrome (MS) (according on the NCEP ATPIII directive) were fulfilled. existent item. All these data militates in favour of a single network that process Results: Of the 64 individuals, MONO was identified in 3.12%, MOO 17.19%, homeostatic and hedonic data. MHO 31.25% and MHNO 48.44% of the subjects. The prevalence of the metabolic disorders among the MONO, MOO was significatively higher when compared to the MHO and MHNO subtypes: fasting hyperglycemia, type 2 diabetes, 16-01 dyslipidimia (mostly type IV and IIb of the Frederickson classification), presence of Lp(a) and elevated ratio triglyceride/HDL (index of small dense LDL), insulin Cell signaling and lipid taste perception in mice: Role of MAPK a a b a c resistance and hyperuricimia. Metabolic obesity was strongly correlated to waist S Abdoul-Azize , S Subramaniam , G Dramane , AM Simonin , H Sadou , adiposity. P Besnarda, N Khana aPhysiologie de la Nutrition & Toxicologie (NuTox), UMR b Conclusion: Our findings reinforce the notion of ‘healthy’ obesity and cast doubt INSERM U866, AgroSup/Universite de Bourgogne, Dijon, France; Labratoire de on the use of BMI on its own to decide on whether to look for MS. Biochimie, Universite de Parakou, Parakou, Benin; cLaboratoire de Nutrition, References: Universite Abdou Moumouni, Niamey, Niger Lacobellis G., Ribaudo M.C., Zappaterreno A. et al. Prevalence of uncomplicated Recent studies in rodents and humans suggest the existence of a sixth taste obesity in an Italian obese population. Obes. Res. (2005) 13 1116–1122. modality: the taste of fat. CD36, a glycoprotein expressed predominantly by the Loganathan Geetha B.D.S., M.P.H. Prevalence and Clinical Profile of Metabolic papillae of the tongue is involved in sensory perception oro-fat food. Regarding Obesity and Phenotypic Obesity in Asian Indians. J Diabetes Sci Technol. (2011) 5 the mechanisms of signaling in mice, linoleic acid, a long chain fatty acid (LCFA) 2+ 439–446. induces rapid increase in intracellular calcium [Ca ]i in taste cells expressing Derguine R., Yargui L., Bouali F., et al. La dyslipidemie de l’obese en bonne sante CD36. This phenomenon is involved in neurotransmitter release from these cells. apparente: rapports avec l’insulinoresistance. Congres SNFMI. (2008), Luxem- In this study, we have deepened the involvement of signaling cascade by study- bourg. ing the activation of MAP kinases. Upon activation of CD36 cells in mouse cir- Keywords: Obesity, metabolic syndrome, insulin resistance cumvallate papillae we found that LCFA induces the activation cascade of MEK1/ 2, ERK1/2, ELK-1. This fatty acid also induces the phosphorylation of MAP kinases p38 and JNK. In addition, we show that the MAPK kinases ERK is coupled to the release of serotonin to promote lipid preference. Our results suggest, for 16-04 the first time, the importance of MAP Kinases phosphorlylation in lipid taste per- Oxidative stress and metabolic syndrome: interest of manganese and chromium evaluation in Algerian type 2 diabetes patients ception. It now remains to consider all the components that play a role in a b b c d c upstream and downstream MAPK cascades. Comprehensive knowledge of all H Harani , A Otmane , M Makrelouf , N Ouadahi , B Alamir , A Berrah ,A Zenatie, NA Khanf, E Koceirg aUSTHB, FSB, ALGER; bLaboratoire Central, Unitede pathways bet in play in this perception could help fight against obesity by devel- c oping molecules like fatty acids that may mimic their effects, or by developing Biochimie, CHU de Bab El Oued, Alger; Service de Medecine interne, Unite d’exploration des maladies metaboliques, CHU de Bab El Oued, FacultedeMedecine various pharmacological molecules to reduce the attraction to foods high in fat. d Keywords: MAP kinases, gustatory cells, lipids, CD36, gustation d’Alger, Alger; Laboratoire de Toxicologie, Unite d’exploration des oligoelements, CHU de Bab El Oued, FacultedeMedecine d’Alger, Alger; eLaboratoire Central, Unitede Biochimie, CHU de Bab El Oued, FacultedeMedecine d’Alger, Alger; fPhysiologie de la Nutrition & Toxicologie, INSERM UMR866, AgroSup/UB, Dijon, France; gLaboratoire de Bioenergetique et Metabolisme Intermediaire, LBPO, Alger In type 2 diabetes, the relationship between antioxidants and insuline-like trace elements is very complex during oxidative stress, being mediated by hyperglycemia, dyslipidemia and inflammation. We investigated the antioxidant status, particularly Mn and Cr on the diabetes metabolic control, and their interaction with

the metabolic syndrome (MS) parameters. The study was undertaken on 278 16-07 Algerian diabetic subjects who were divided in two groups according to glycated Viral hepatitis is aggravated by TLR-2 dependent increases of hemoglobin (HbA1c) <7% or >7% value, attesting for a good or poor metabolic inflammatory chemokines and cytokines in a murine model control of diabetes, respectively. The MS was defined according to NCEP-ATPIII. C Bleaua, M Burnettea, M Samsonb, L Lamontagnea aUniversiteduQuebec a Insulin resistance was evaluated by HOMA-IR model. The plasma manganese Montreal, Montreal, QC, Canada; bUniversite de Rennes 1, Rennes, France concentrations were significantly increased in both diabetics groups, indepen- Objective: Viral sensors on inflammatory cells detect viral products that trigger dently of metabolic control. However, chromium (Cr) seems to play a determi- innate antiviral responses. While most of these sensors are intracellular and rec- nant action in metabolic control, as shown by better values of insulin resistance ognize viral genome, recent results suggest that surface sensors such as TLR-2 (HOMA-IR) and HbA1c. The Selenium status was positively correlated with and -4 may also participate through ligation of viral coat proteins. It was glutathion peroxidase activity. Copper and Zinc plasma levels in the diabetic recently reported that HCV could activate TLR2-mediated innate immune signal- patients were similar to those of control subjects. In conclusion, our results sug- ing but the consequences on the pathogenesis of hepatitis remains unclear. The gest that Mn play a crucial role in antioxidant capacity and we hypothesize that objective of this study was to evaluate the role of TLR2 in a murine model of antioxidant defense is preserved in the cytosol (superoxide dismutase Cu/Zn- viral hepatitis. SOD), whereas it is impaired in mitochondria (Mn-SOD), which makes this cell Material and methods: C57BL/6 mice were infected with a high-, mild- or low- organelle a true therapeutic target in diabetes. pathogenic mouse hepatitis virus (MHV) (L2-MHV3, MHV-A59 and YAC-MHV3 Keywords: type 2 diabetes, trace elements, manganese, chromium, insulin resis- respectively) and sacrificed 24 h, 48 h and 72 h pi. TLR-2 À/À mice were also tance, oxidative stress infected with L2-MHV3 and sacrificed at the same times. Liver was extracted and samples were frozen for RNA and protein extraction or fixed in formaldehyde for histopathological analysis. Blood was collected for serum determination of ASAT 16-05 and ALAT levels. Liver expressions of TLRs, helicases, cytokines and chemokines Effects of Zizyphus lotus L. (Desf.) polyphenols on Jurkat cell signaling were analyzed by RT-qPCR and/or ELISA. and proliferation Results: As most of TLRs and helicases levels were higher in liver after infection M Bendahmanea, S Abdoul-Azizeb, A Hichamic, G Dramaned, AM Simonine, with both mild- and high-pathogenic MHV-A59 and L2-MHV3 viruses. TLR-2 N Khane aLaboratoire de Biologie de Reproduction, Universite de Sidi Bel-Abbes, Sidi expression, however, increased radically following pathogenic L2-MHV3 infection Bel-Abbes, Algerie; bINSERM U866, Physiologie de la Nutrition & Toxicology in comparison to other MHVs. Levels of the cytokines IL-6 or TNF-a and the (NuTox), Universite de Bourgogne, Dijon, France; cChimiotherapie, metabolisme des chemokines CXCL1, CCL2/3 and CXCL10/11 were also higher in livers from lipides et reponse immunitaire anti-tumorale INSERM U866, Dijon, France; pathogenic virally-infected mice and correlate with the MHV pathogenicity. These dLabratoire de Biochimie, Universite de Parakou, Parakou, Benin; eINSERM U866, levels were however significantly lower in livers of TLR2 À/À mice and were Physiologie de la Nutrition & Toxicology (NuTox), Universite de Bourgogne, Dijon, associated to a less extensive necrosis and fewer inflammatory foci when com- France pared to livers from L2-MHV3-infected C57BL/6 mice. Zizyphus lotus L. (Desf.) is a medicinal plant used in traditional medicine for its Discussion: These results suggest that the pathogenicity of MHVs may be related multiple therapeutic properties and consumed for its nutritional values. We to their capacity to induce TLR-2 expression leading to an exarcebate TLR-2- assessed the effects of Zizyphus lotus L. (Desf.) polyphenols (ZLP) on T-cell signal- mediated inflammatory response. Blocking of TLR2 signaling cascade may be ing and proliferation. Our results showed that ZLP exerted no effect on the considered as a potential therapeutic approach in controlling viral-induced 2+ inflammatory diseases such as viral hepatitis. increases in intracellular free calcium concentrations, [Ca ]i, in human Jurkat 2+ Keywords: hepatitis, virus, TLR, chemokines, cytokines T-cells. However, ZLP modulated the thapsigargin-induced increases in [Ca ]i in these cells. ZLP treatment was found to decrease the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2). In addition, ZLP induced a rapid (t½=33 s) and dose-dependent decrease in intracellular pH (pHi) in human Jur- 16-08 kat T-cells. Furthermore, ZLP significantly curtailed T-cell proliferation by dimin- Acoustic signal analysis at esophagogastric junction during swallowing: ishing their progression from S to G2/M phase of cell cycle, and the expression of comparison between healthy subjects and patients with interleukin-2 (IL-2) mRNA. Taken together, the results of the present study dem- gastroesophageal reflux disease onstrate that ZLP modulate cell signaling and exert immunosuppressive effects in KG Soroa, D Jauneaub, E Parmentierb, Z Benchellalc, N Hutenc, human T-cells. M Boironb aService de Chirurgie Generale et Digestive - CHU de Yopougon., Abidjan, Keywords: Zizyphus lotus L. (Desf.) polyphenols, Jurkat T-cells, intracellular pH, Côte d’Ivoire; bLaboratoire Universitaire de Physiologie et Motricite Digestive - UFR calcium, IL-2, cell cycle. Medecine - 37032, Tours, France; cService de Chirurgie Viscerale – CHRU Trousseau – 37044, Tours, France Background: Swallowing sounds can be heard in upper esophagus by cervical 16-05 auscultation (1–3) and also in lower esophagus at the esophagogastric junction Nutritional survey of iron deficiency anemia in women of two regions of (EGJ). This lower swallowing sound is composed of vibration groups separated by southern Algeria period >100 ms (4–5). FZ Abi-Ayada, D Lakhdarb, M Abi-Ayadc, F Abdib aUniversite de Mascra- Universite Aim: The aim of this study was to compare the swallowing xiphoid sound analy- de Tlemcen, Tlemcen, Algerie; bUniversite de Mascara, Mascara, Algerie; cUniversitede sis in healthy subjects and gastroesophageal reflux disease (GERD) patients in Tlmecen, Tlemcen, Algerie physiological conditions. Aim: Anemia is defined by the decrease in circulating hemoglobin mass, there Method: Twelve healthy subjects and 14 patients with GERD were included in are several types of anemia according to the causes and among them iron defi- the study. Two microphones were placed below the cricoid and on the xiphoid ciency anemia. The aim of our study is to search if the tea is a chelator of iron, cartilages. The later sound is called ‘xiphoid’ sound. The swallowing sound activ- leading to anemia. ity was transferred to computer for subsequent quantitative analysis performed Patients & Methods: To determine the impact of iron deficiency anemia, we under a specific software ‘Cool Edit Pro’ program. Each subject was asked to per- focused our study on women of two regions of Algeria: 34 cases of the city of Be- form a six-swallow sequence with 10 ml of water. Six acoustic parameters were char and 40 cases of the city of Adrar. calculated: total duration of xiphoid sound, number of vibration groups, vibration To study this type of anemia, we performed hematological tests as blood counts, group duration, interval between groups, number of vibrations per group and biochemical tests and blood smears. interval between vibrations. A specific software was developed for automatic Discussion: We made a comparison between the two regions and we found that analysis. All values were expressed as means with SD. The Mann–Whitney test the cause of iron deficiency anemia in Adrar and Bechar is the abuse of con- was applied for the statistical analysis. sumption of tea (the rate of consumption and cooking time). Results: Total duration of xiphoid sound and Number of vibration groups were Keywords: iron deficiency anemia, tea, hematological tests significantly increased in patients compared to healthy subjects. Duration of vibration groups and number of vibration per group was significantly lower and interval between groups was significantly higher in patients compared to healthy 16-06 subjects. An acoustic profile at the EGJ in healthy subjects and patients was established. Impact of food fats on the taste perception in obese people a a Discussion: This study allowed us to describe the main variations of the xiphoid I Karmous faculte des sciences de Bizerte, Bizerte, Tunisie sound induced by GER. These evaluations should provide information on the pas- Aims: To test the lipid taste perception in obese people compared to a control sage modalities of the bolus at the EGJ. This non invasive and physiological group. method could be performed after GERD surgical treatment. Population: This study concerned a population of 100 persons of both sexes, References: non-smokers, without diabetes or hypertension, aged between 30 and 50 years 1. Moriniere S., Boiron M., et al. Dysphagia (2008) 23 267–273. and divided into two groups: 50 obese (BMI > 35) and 50 non-obese controls – Æ 2. Moriniere S., Boiron M., et al. Dysphagia (2011) 26 366 373. (BMI 22). 3. Baulieu F., Boiron M., et al. Dysphagia (2007) 22 281–289. Methods: The following tests were realized: 4. Boiron M., Benchellal Z., et al. J Gastrointest Surg (2009) 13 1570–76. An anthropometric test to measure weight, size and eating habits food (food 5. Boiron M., et al. Dis Esophagus (2009) 22 68–73. questionnaire and survey) Keywords: swallowing sounds - esophagogastric junction - physiological condi- Taste perception test based on linoleic acid which are prepared at different con- tions – acoustic centrations (0.018 mM–12 mM) Serum leptin, CCK, glucose, cholesterol, HDL-cholesterol, LDL-cholesterol, triglycerides, hemoglobin, creatinine, uric acid, NFS, VS, ALAT and ASAT were measured at the beginning and at the end of the test. 16-09 Results: The obese group presented a low taste perception. In this group, a gus- Effect of fatty acids of nigella sativa L. seeds oil in the prevention of the tative perception is observed only in strong concentrations. insulinoresistance and the hyperlipidemia on obese male rat a a a a a a The control group presented a good food fat perception even for a low lipid con- A Didi , A Benhamidat , RK Meziane , F Amamou , D Chabane Sari universite centration. de Tlemcen, Tlemcen, Algerie Conclusion: The lipid gustative perception is more developed at not obese per- Obesity becomes the lead health problem in the 21st century, and consumption sons compared with the obese group. of energy-dense foods rich in fat and carbohydrate is considered an important Keywords: obesity, gustation, lipids, fat contributing cause [1]. Recent researches show a preventive effect for obesity with supplementary by polyunsaturated fatty acids [2]. In this aim, our study

explore the effect of addition in high-fat diet of Black seeds fixed oil (Nigella sativa 16-12 L.) which is rich with polyunsaturated fatty acids (88.5%) [3] and estimate their Cocoa and health: a study of the cardioprotective effect of cocoa powder impact in preventing the installation of insulin resistance and obesity in rats. on women with hypertension and menaupose For this three (03) groups of mal albinos Wistar rats (1 mouth old) during eight F Abdia, D Lakhdara, K Ibria aUniversite de Mascara, Mascara, Algerie weeks were fed different diets (n = 5); group 1: standard diet (with 4% sunflower Aim: Epidemiological studies show that regular consumption of plant foods oil), group 2: High fat diet (32% sunflower oil) and group 3: High fat diet treated reduces the risk of cardiovascular disease. In particular, consumption of cocoa diet (28% sunflower oil+4% nigel oil). Our results show a high significant powder has many beneficial effects on health. The aim of our study is to supple- decrease in body weight during the eight (08) weeks, a high correction in blood mentation of cocoa powder (5 g and 10 g) in the diet of a sample of individuals parameters (glucose, triglycerids and total cholesterol levels). Also, results show with hypertension. significant decrease in insulin blood concentration in group 3. Population & Methods: Our study involved 67 people with hypertension, aged So, addition of Black seeds oil play an important role on some metabolic compli- 45–78ans which the majority is female (10 men and 57 women), classified cations thus it seems interesting to continue this study with further investiga- according to age and followed or not hypotensive medical treatment. Nutritional tions. quality, microbiological and physicochemical cocoa powder were analyzed by sev- References: eral techniques. Lipid profile (total cholesterol, TG, HDL-C and LDL-c), blood glu- 1. Hong J.I., Friedman M.I., Reduced capacity for fatty acid oxidation in rats with cose were measured by biochemical methods, the blood pressure and inherited susceptibility to diet-induced obesity. Metabolisme Clinical and experi- anthropometric parameters were performed. mental (2007) 56 1124–1130. Results: The results obtained on supplementation of a diet with cocoa powder 2. Koji N., Teruyoshi Y. Bioactive lipids in metabolic syndrome. Progrs in Lipid are consistent with the literature data. It contributes to the reduction of blood Res (2007) 47 127–46. pressure, blood glucose, total cholesterol, triglycerides and LDL-c, and increased 3. Ramadan M.F., Moersl J.T., Screening of the anti-radical action of vegetable HDL-c is a cardioprotective factor. oils. Journal of food composition and analysis. (2006) 19 838–842. Discussion: These beneficial effects of cocoa powder are associated with its com- Keywords: fatty acids, insulin, lipid profil, obesity, rat ponents: its high content of polyphenols, methylxanthines (theobromine, phenyl- ethylamine …) and trace elements (iron, magnesium, zinc …). Keywords: cocoa polyphenols, hypertension, HDL-c, LDL-c, theobromine, cardio- 16-10 vascular disease, diet Antifibrosis effects of All-trans-Retinoic Acid in a rat model of bleomycin-induced pulmonary fibrosis R Serairi Bejia, A Abidib, R Zemnib, S Ben Khamsa Jameleddineb aEcole Superieure des Sciences et Techniques de la Sante de Tunis, Tunis, Tunisia; bFacultedeMedecine de SESSION 17: GS SOMMEIL Tunis, Tunis, Tunisia Introduction: All-trans-retinoic acid (ATRA), a physiologic metabolite of vitamin Intermittent hypoxia and metabolic consequences A, is known to affect cell differentiation, proliferation, and development. Pulmo- a a nary fibrosis is one of the most common chronic interstitial lung diseases with F Gagnadoux Departement de Pneumologie, Centre de Medecine du Sommeil, CHU, high mortality rate after diagnosis and limited successful treatment. Angers INSERM 1063 SOPAM, Angers, France Methodology: The present study was designed to assess the potential antifibrotic Intermittent hypoxia (IH) occurs during sleep in patients with obstructive sleep effect of ATRA and whether ATRA can attenuate the severity of oxidative stress apnea (OSA), as an immediate consequence of recurrent episodes of partial or and inflammatory response during bleomycin-induced pulmonary fibrosis in rats. complete obstruction of the upper airways. There is growing evidence in support Treatment groups included: (i) a single intratracheal (i.t.) instillation of bleomycin of an independent association between OSA and metabolic dysfunction including and weekly intraperitoneal (i.p.) injection of 3 mg/kg ATRA; (ii) i.t. bleomycin impaired glucose metabolism and dyslipidimia. Type 2 diabetes and metabolic syn- and i.p. diluent (cottonseed oil); (iii) i.t. saline and i.p. ATRA, 3 mg/kg weekly, drome are common comorbid conditions in patients with OSA. An independent (iv) i.t. saline and i.p. diluent. Eight rats in each group were sacrificed on day 28 association has been demonstrated between sleep-disordered breathing severity after intratracheal instillation. Fibrosis was assessed by measuring fibrosis score and poorer glucose control as assessed by HbA1c in both diabetic and non-dia- and inflammatory index. Levels of lipid peroxidation in lung were estimated by betic subjects. Population based studies in middle-aged adults have demonstrated measuring the formation of thiobarbituric acid reactive substances (TBARS). that intermittent nocturnal hypoxia is associated with insulin resistance, glucose Superoxide-dismutase (SOD) activity was determined in lung homogenate using intolerance and with a higher risk of developing diabetes. Chronic IH could poten- the method of Beyer and Fridovich. Catalase (CAT) activity was measured accord- tially be detrimental to glucose metabolism via intermediate mechanisms including to the method of Aebi and glutathione-peroxidase (GPX) activity was mea- ing activation of the sympathetic nervous system, hypothalamic-pituitary axis and sured using the method of Flohe and Gunzler. inflammatory pathways. IH results in insulin resistance in both genetically obese Main Results: Both fibrosis score and inflammatory index are increased mark- and lean mice. In healthy subjects, sustained hypoxia acutely induces glucose edly in the bleomycin group 2 but and significantly reduced by concurrent treat- intolerance and increased plasma epinephrine concentration. Dyslipidemia may ment with ATRA (group 1). Furthermore, histopathological examination also contribute to the increased cardiovascular risk in OSA. An independent asso- confirmed the antifibrotic effect of ATRA. ciation has been observed between nocturnal intermittent hypoxemia and meta- The levels of TBARS and the GPX activities significantly increased in group 2 bolic dyslipidemia characterized by a combination of increased triglyceride (TG) compared with the values obtained in group 1. ATRA administration to rats trea- levels and low high-density lipoprotein cholesterol levels (HDL-c). Data from ani- ted with bleomycin caused a significant decrease in TBARS production and GPX mal models suggest that IH may disrupt lipid metabolism by increasing adipose activities when compared to rats treated with bleomycin alone (respectively tissue lipolysis and FFAs flux to the liver, by upregulating hepatic TG biosynthesis 2.85 Æ 0.38 vs. 6.04 Æ 0.78 nmol/mg protein for TBARS and 1.47 Æ 0.27 vs. and lipoprotein secretion, and suppressing lipoprotein clearance. Emerging evi- 2.48 Æ0.28 nmol GSH/min/mg protein for GPX). dence also suggests that OSA may play a role in the progression of hepatic steato- The levels of CAT and SOD activities were significantly decreased in the group 2 sis and the development of non-alcoholic steatohepatitis (NASH). Several cross- when compared to the values in the group 1 (respectively P < 0.01 and sectional studies showed that the severity of IH in patients with OSA predicted the P < 0.001). severity of non-alcoholic fatty liver disease (NAFLD) on liver biopsy. In mouse Collectively these findings indicate that ATRA treatment significantly attenuated models, IH induces oxidative stress in the liver and accelerates the progression of the increased pulmonary damage induced by bleomycin. This study supports the hepatic steatosis by inducing adipose tissue lipolysis and increasing free fatty acids use of ATRA to prevent or ameliorate lung fibrosis. flux into the liver. IH also up-regulates HIF-1 alpha and possibly HIF-2 alpha, Keywords: ATRA- pulmonary fibrosis-oxidative stress which may increase hepatic steatosis and induce liver inflammation and fibrosis.

16-11 17-05 Determination of the activities of lipases during the experimental obesity Assessment of impaired in vitro endothelial repair function in M Sari Hassouna, N Mokhtari-Soulimanea, H Merzouka aLaboratory of Physiology, obstructive sleep apnea patients and impact of associated metabolic syndrome Pathophysiology and Biochemistry of Nutrition (PPABIONUT), Tlemcen, Algerie a a a b b Until recently it was thought that obesity is the result of the interaction between A Briancon-Marjollet , M Henri , E Lemarie , JL Pepin , R Tamisier , P Levyb aINSERM U 1042, Laboratoire HP2, Hypoxie : Physiopathologies; Universite an individual’s behavior and its genetic heritage. Recent data indicate that fetal b programming plays an important role in the pathogenesis of this disease. Among Joseph Fourier, FacultedeMedecine, Grenoble Cedex 09, France; INSERM U 1042, Laboratoire HP2, Hypoxie : Physiopathologies; Universite Joseph Fourier; CHU de the pathophysiological mechanisms, epigenetic modifications seem to play a lead- ^ ing role. Grenoble, Pole Locomotion, Reeducation et Physiologie, Grenoble Cedex 09, France In order to evaluate the impact of maternal obesity on lipid metabolism of the off- Rationale: Obstructive Sleep Apnea (OSA) is a major public health concern due spring, we used as an experimental model of nutritional obesity in Wistar rats to its cardiovascular morbidity. Intermediate mechanisms involved are sympa- fed cafeteria diet (Darimont et al., 2004). At J30 and J90 of birth, the offspring thetic activation, oxidative stress and vascular inflammation. All these are were weighed and sacrificed. Blood and organs were collected. Serum glucose, involved in atherosclerosis pathway through an imbalance between vascular cholesterol (CT) and triglyceride (TG) concentrations, total lipid content of injury and repair mechanisms. Little is known about endothelial repair function organs, and the activities of lipoprotein lipase (LPL) and hormone-sensitive lipase in OSA, in which, interestingly have been reported an increase in plasma VEGF. (HSL) of adipose tissue were measured. The aim of this study was to investigate the factors present in human serum Our results indicated that newborns of mothers under cafeteria diet were macro- from 81 OSA patients and controls, with or without metabolic syndrome (MS), that affect in vitro endothelial wound healing and monocyte migration. somic and became obese in adulthood, whatever their postnatal diets. In addition, = at weaning, the offspring born to obese mothers has an accumulation of adipose Methods: We investigated healthy controls (n 16), OSA patients free of any other cardiovascular or metabolic diseases (n = 31), OSA patients with MS tissue with lipids enrichment, associated to an increase in LPL activity and adipo- = = cyte LHS, hyperglycemia and hyperlipidemia characterized by high serum and (n 21) and non-OSA patients with MS (n 13). In all subjects serum, VEGF lipoprotein CT and TG and low HDL-C levels. These metabolic alterations were and usCRP levels were assessed and their specific function were tested with or worsened by rat age regardless to the postnatal diet. without antibodies blocking VEGF (anti-VEGF) or CRP (anti-CD32a) on endothe- In conclusion, the programming effects of maternal obesity during pregnancy lial cells (HMVEC) for wound healing assays and on human monocyte cell line exert permanently long-term changes in the offspring that contribute to the (THP-1) for Transwell migration assays. maintenance of obesity. Results: Along with the presence of OSA and/or MS, both VEGF and usCRP Keywords: obesity, cafeteria diet, LPL, LHS, Wistar rats were significantly elevated. Compared to controls, endothelial wound healing was

impaired when HMVECs were incubated with OSA patient serum, and further 17-02 inhibited with OSA+MS patient serum. Blocking VEGF both in OSA and OSA+MS Loss of Restorative properties of sleep in multiple sclerosis fatigue serum revealed an impaired endothelial wound healing. Partial normalization of A Bridouxa,ACreangeb, S Ayachea, H Hosseinib, A Noroca, W Fahrata, endothelial wound healing was resolved when blocking CRP using anti-CD32a JP Lefaucheura, X Drouota aService d’explorations fonctionnelles, hôpital henri antibody. Mondor, Creteil, France; bService de Neurologie, hôpital Henri Mondor, Creteil, France THP-1 monocyte migration was activated by incubation of cells with serum from Rationale: Multiple sclerosis (MS) patients experience constant, severe and post- OSA patients, and further enhanced by serums from OSA+MS patients. Blocking exercise, fatigue (Iriarte et al. 2000). Disturbed sleep is common in patients with CRP in serum inhibited this migration. MS and is likely to contribute to greater fatigue. Nevertheless, the subjectively Conclusions: Serum from OSA patients impairs endothelial cell repair function assessed fatigue is not correlated with severity of disturbed sleep (Kaynak et al, and monocyte migration in vitro, which are aggravated in OSA patients with 2000). The modulation of motor cortex excitability after exercise can be evalu- MS. These effects are partly driven by VEGF and usCRP, although other factors ated using transcranial magnetic stimulation (TMS). A fatiguing exercise induces may also be involved. This suggests that the unfavorable balance between pro long lasting depression of motor-evoked potentials (MEP). This depression is healing and pro injury factors may promote atherosclerosis, and that VEGF might accompanied with central neurological fatigue and reflects objectively the fatiga- be a positive factor counteracting the deleterious effects of CRP. bility following an effort (Zanette et al, 1995). Our hypothesis proposes that Keywords: Sleep Apnea, endothelial repair, monocyte activation, Vascular Endo- restorative properties of sleep have beneficial effect on motor recovery after exer- thelial Growth Factor, CRP cise in healthy subjects whereas no restorative property of sleep in MS has an impact on motor fatigability. Objective: To compare the effect of post exercise nap vs. rest on the recovery of 17-06 MEP amplitudes with TMS and central neurological fatigue in MS patients and In sleep apnea patients nonalcoholic fatty liver disease (NAFLD) is healthy subjects. associated with the severity of intermittent hypoxia and more severe Methodology: Serial MEP recordings were performed from the thenar eminence endothelial dysfunction before, 10 and 60 min after an exercise of thumb contraction at 25 % of maxi- C Minvillea, MN Hilleretb, R Tamisierc, P Levyc, JP Zarskib, JL Pepinc aInstitut mal force for 24 min. The morning, after the exercise, the subject is maintained universitaire de cardiologie et de pneumologie de Quebec, QC, Canada; bDepartment of at rest. The afternoon, the same protocol has been applied but the rest period Endocrinology, Pôle Digidune, Grenoble University Hospital, Grenoble Cedex 09, was switched for a nap. We compared the difference of MEP amplitudes 10 and France; cINSERM U 1042, HP2 Laboratory (Hypoxia: Pathophysiology), Joseph 60 min after the exercise in the two conditions: at rest and after a nap in healthy Fourier University and Sleep Laboratory and EFCR, Locomotion, Rehabilitation and subjects and in MS. Physiology Department, University Hospital, Grenoble Cedex 09, France Results: We found a 70% decrease of MEP amplitudes after exercise. MEP ampli- Introduction: Nonalcoholic fatty liver disease (NAFLD) begins with the accumu- tudes were significantly increased after nap comparing to rest in healthy subjects lation of triglycerides in the liver and elicits an inflammatory response that can (P 0.05). In MS, there were no significant differences in MEP amplitudes progress to cardiovascular complications, cirrhosis and liver cancer. Intermittent recording between nap and rest. hypoxia is a potential contributing factor but NAFLD has not been investigated Discussion: Our results confirm our hypothesis and show that sleep has benefi- in an unselected obstructive sleep apnea (OSA) population. Beyond liver biopsy, cial effect on motor recovery in healthy subjects and no beneficial effect in MS. there are non invasive validated tools allowing a screening of NAFLD in large The fatigue management has been largely disappointing. A better understanding populations. of the pathophysiology of fatigue and its relationship with sleep in MS are Aims: (i) To use non-invasive blood tests (Steatotestâ, NASHtestâ and Fibrotestâ) needed. to evaluate steatosis, Nonalcoholic Steato hepatitis (NASH) and fibrosis in a large Keywords: multiple sclerosis- fatigue -sleep- fatigability post-exercise cohort of OSA (II) To assess endothelial function by peripheral arterial tone (PAT). Patients: Two hundred and twenty-six subjects referred for suspicion of OSA 17-03 were included (men: 55%, median age: 56 years, mean BMI: 34 kg/m2). Nocturnal desaturations in sickle cell disease: Polysomnographic study Results: 61.5% of OSA patients exhibited advanced steatosis. By multivariate in adults analysis, triglycerides (P < 0.0001), insulin resistance (P = 0.0004) and noctur- A Habibia, A Covali Norocb, D Bachira, A Bridouxb, B Maitreb, F Galacterosa, nal cumulative time spent <90% of SaO2 (CT90) (P = 0.01) were independent L Boyerb, S Adnotb, X Drouotb aAPHP Groupe Hospitalier Henri Mondor Centre factors for liver steatosis. 38% of OSA displayed NASH (N1 or N2 with NASH- Reference Syndrome Drepanocytaire majeur, Creteil, France; bAPHP Groupe Hospitalier testâ). CT90 was significantly associated with NASH (P = 0.035) but this became Henri Mondor Service de Physiologie, Creteil, France non significant in multivariate analysis. Endothelial function was more impaired Introduction: Sickle cell disease (SCD) is an inherited blood disorder producing in OSA patients with advanced steatosis (P = 0.04) and NASH (P = 0.013). sickle hemoglobin (HbS). HbS polymerizes when it releases oxygen to tissues, Discussion/Conclusion: In a large unselected population of OSA, the severity causing the normally flexible red blood cells to become rigid and to obstruct of intermittent hypoxia was independently associated with steatosis. Endothelial blood flow. The subsequent ischemia results in acute organ damage called vaso- dysfunction was more severely impaired in OSA patients demonstrating NAFLD. occlusive crisis (VOC). Hypoxia is considered as a major factor in VOC. In some Keywords: Nonalcoholic steatohepatitis, steatosis, metabolic liver disease, non SCD patients, VOC frequently occur during sleep. In children, nocturnal desatura- invasive biomarkers, Fibromax tion episodes are frequent and are associated with an increased frequency of VOC. Several studies reported high prevalence of obstructive sleep apnea syndrome in children. A mean nocturnal saturation below 96% predicts central ner- 17-01 vous event such as strokes. In addition, adenotonsilectomy in children reduced Relationship between Sympathetic Activity and early cardiovascular the number of cerebrovascular ischemia. These studies suggest that nocturnal de- disease in ‘healthy’ OSA saturations associated with obstructive sleep apnea syndrome may play a major R Tamisiera, D Lacedoniab, JP Baguetc, JL Pepina, P Levya aINSERM U1042,HP2 role in VOC. In SCD adults, the role of sleep apnea in the occurrence of nocturnal Laboratory (Hypoxia: Pathophysiology); Joseph Fourier University; Locomotion, VOC is unknown. Rehabilitation and Physiology Department, University Hospital, Grenoble Cedex 09, Our hypothesis was that nocturnal VOC in adults could be linked to desaturation France; bINSERM U1042,HP2 Laboratory (Hypoxia: Pathophysiology), Joseph episodes secondary to sleep apnea syndrome. Fourier University; Institute of Respiratory Diseases, Department of Medical and Methods: We recorded sleep with polysomnography in consecutive SCD adults. Surgical Sciences, Univerity of Foggia, Foggia, Italy; cCardiology Department, Polysomnographies were justified because of frequent nocturnal VOC, snoring, University Hospital, Grenoble Cedex 09, France sleep symptoms such as sleepiness or non restorative sleep. Standard PSG were Rationale: Obstructive sleep apnea syndrome (OSAS) is associated with high used with digital pulse oxymetry. Sympathetic Activity (SA) and a subsequent risk of hypertension and atheroscle- We compared PSG of patients reporting predominant nocturnal VCO with PSG of rosis. Objectives of this study were to assess SA and its impact on early cardiovas- patients reporting predominant diurnal VOC. cular disease markers in OSA. Results: Among the sixty-three patients recorded, 29 (46%, six females) reported Methods: We studied 55 subjects with suspect of OSA and controls, who were predominant nocturnal VCO. Apnea – hypopnea index (5/h, 2–11 vs. 4/h, 0–8, otherwise healthy, without evidence of clinic hypertension. SA was measured by P > 0.05, Mann–Withney) and desaturation index were not significantly different direct peroneal microneurography: Muscle Sympathetic Nerve Activity (MSNA) in both groups. In contrast, time spent below 90% was higher in patients with and all subjects underwent a full polysomnography, 24 h Ambulatory Blood predominant nocturnal VOC (0 mn, 0–1 vs. 6 mn, 0–77, P < 0.05, Mann–With- Pressure Monitoring (ABPM), arterial stiffness by Pulse Wave Velocity (PWV), ney) and minimal saturation and mean saturation during sleep were significantly vascular reactivity by Peripheral Arterial Tone (PAT) and early signs of athero- lower in patients with predominant nocturnal VOC. sclerosis by arterial carotid Intima Media Thickness (IMT). Discussion: Sleep apnea syndrome does not seem to be a major factor for noc- Results: Thirty-eight OSA patients and 17 aged-matched controls were enrolled. turnal VOC in SCD adults. On the other hand, low nocturnal saturations could Both were comparable for all but not for BMI (25.92 Æ 2.71 vs. favor nocturnal VOC. Pathophysiology of these low saturation remains to be elu- 24.20 Æ 2.39 kg/m², P = 0.02). Systolic and diastolic ambulatory blood pressure cidated. were the same in patients and controls, while on ABPM 24 h-diastolic pressure Keywords: sickle cell disease; sleep apnea; nocturnal desaturation; polysomnog- was higher in OSAS group (79.92 Æ 8.73 vs. 74.59 Æ 8.83, P = 0.046). No dif- raphy ferences were found about PAT, PWV and IMT. At baseline SA was higher in OSA than in controls 39.71 Æ 9.55 vs. 31.10 Æ 8.00 bursts/min, P < 0.01. There was a strong correlation between SA and all blood pressure parameters: 17-04 systolic and diastolic office pressure (R = 0.39 and 0.42, P < 0.01) and systolic Transcriptional leukotriene pathway activation in Apolipoprotein-E and diastolic 24-ABPM (R = 0.38 and 0.46, P < 0.001), as well as between SA deficient mice exposed to intermittent hypoxia: role in atherosclerosis and PWV (R = 0.28, P < 0.05). Preliminary data on 15 OSA and nine controls progression were obtained after 6 month and show that CPAP treatment reduced SA in OSA E Gautiera,MBack€ b, C Arnauda, M Petrib, P Levya, F Stanke- patients (Bursts/min 40.49 Æ 10.63 vs. 35.47 Æ 10.06, P < 0.05). Labesquec aLaboratoire HP2, INSERM, U1042, Grenoble, France; bDepartment of Conclusion: In ‘Healthy’ OSAS SA is likely to play a significant role in the path- Medecine, Karolinska Institue and University Hospital, Stockholm, Sweden; ogenesis of hypertension and atherosclerosis. Use of CPAP, by reduction of SA, cLaboratoire HP2, INSERM, U1042, LA Tronche, France may reduce the risk of future development of cardiovascular diseases. Aim: 5-lipoxygenase (LOX)-pathway is activated in obstructive sleep apnea syn- Keywords: Sympathetic activation, Sleep apnea, Blood pressure, Hypoxia drome (OSA) and seems to be involved in the early vascular remodelling associated to OSA1. Exposure of Apolipoprotein-E deficient (ApoEÀ/À) mice to

intermittent hypoxia (IH) accelerated atherosclerosis progression2 but the link disorders in attentional reactivity, particularly in response to emotional stimula- between IH, atherosclerosis and LOX-pathway remains to be further explored. tion. Some works report biases for processing positive and negative stimuli in The aim of this study was to investigate the transcriptional profile of the LOX- patients with mania and depression. To evaluate these biases, most studies use pathway and its link with atherogenesis in ApoEÀ/À mice exposed to IH. either a subjective approach (with a verbal assessment of the valence of a stimu- À/À Methods: 40 ApoE mice were exposed to IH (21–5% FiO2, 60 s cycle, 8 h/ lus) or an objective approach (measurement of electrodermal response). Both of day) or normoxia (N) for 8 weeks. Atherosclerosis on aortic roots was quantified these approaches have an influence on the response and may cause methodologi- by Oil-Red-O and genomic expression of LOX-pathway and cytokines were evalu- cal bias. For exemple, it was found that verbal response could be influenced by ated in aortas by real time quantitative reverse transcriptase-PCR. social desirability and the electrodermal response could be influenced by electrode Results: IH aggravated atherosclerosis (P = 0.008) without change of plasma contact. There is a need to measure physiologically attentional response to emo- lipid levels. Genomic expression of several enzymes and receptors of the LOX- tional stimulus with objective and non contact method. Recently, studies have pathway were increased (P < 0.05) in aortas of IH mice (5-lipoxygenase, 5-LOX; proposed pupillometric measurement to assess change in cognitive load (pupillary 5-lipoxygenase activating protein, FLAP; leukotriene A4 hydrolase, LTA4H; cys- psychosensorial reflex) which varies with the mobilization of attentional teinyl-leukotriene receptor 1, Cys-LTr1). Atherosclerosis lesion sizes significantly resources. Pupil size variation can be easily measured with non contact eye correlated to mRNA levels of LOX-pathway genes (5-LOX: r = 0.552, P = 0.05; tracking systems. Similarly, these systems provide access to the measurement of FLAP: r = 0.714, P = 0.01; LTA4H: r = 0.582, P = 0.04; Cys-LTr1: r = 0.543, another physiological variable which changes with attentional processes: blinks P = 0.05) in IH mice; whereas, lesion sizes of N mice only correlated to choles- eyelid. The aim this study was to determinate the relevance of these parameters terol levels (r = 0.571, P = 0.02). Furthermore, RT-PCR data showed greater in response to emotional visual stimulation in healthy adults. Using a non con- mRNA levels of pro-inflammatory cytokines (interleukin 6: P = 0.04; chemokine tact eye tracking system in 22 adults (13 males at an age of 21–39 years ligand 3: P = 0.05) and cytokine receptors (tumour necrosis factor receptor (25.1 Æ 6.61) and nine females at the age of 21–38 years (27.2 Æ 4.71)), pupil (TNFr) 1 and 2: P = 0.02 for two genes) in IH mice than N. size and blinks frequency were measured in response to IAPS pictures (pleasant Conclusion: Exposure of ApoEÀ/À mice to IH accelerated the atherogenesis and or unpleasant). We found an increase of pupil size in reponse to unpleasant stim- activated the transcription of the LOX-pathway, both being highly correlated. uli compared to pleasant stimuli. No difference was observed concerning blinks Since the LOX-pathway also activated the pro-inflammatory cytokine pathway, frequency. Pupillary psychosensorial reflex constitutes good indicator of atten- these data collectively highlight the interest of investigating the effect of leukotri- tional recruitment and would be relevant biomarker to study attentional biais in ene pathway inhibitor in atherosclerosis progression in ApoEÀ/À mice exposed to response to emotional stimuli. IH. Keywords: pupil, blinks, attention, emotion References: 1. Stanke-Labesque et al, J Lipid Res (2012) 1944–1951. 2. Gautier et al, Eur Respir J (2012), in press. 18-02 Keywords: intermittent hypoxia, atherosclerosis, leukotriene Age-related audiovisual interaction changes in the superior colliculus of the rat M Costaa, M Picheb, F Leporec, JP Guillemotd aCERNEC, Departement de Psychologie, Universite de Montreal, Montreal, QC, Canada; bDepartement de SESSION 18: GS MIXTE NEURO chiropratique, UniversiteduQuebec a Trois-Rivieres, Trois-Rivieres, QC, Canada; cDepartement de Psychologie, Universite de Montreal, Montreal, QC, Canada; dDepartement de Kinanthropologie, UniversiteduQuebec a Montreal, Montreal, QC, Role of the subthalamic nucleus in the control of action Canada P Sauleaua, F Le Jeuneb, S Drapiera, JF Houvenaghela, T Dondainea, S Argauda, a a Objective: It is well established that multisensory integration in the superior col- MVerin “Behavior and Basal Ganglia” research unit (EA 4712), University of liculus (SC) disambiguates external stimuli and therefore enhances speed orienta- Rennes 1, University Hospital of Rennes, 9 Rue Jean Mace, Rennes, France; b tion reactions towards simple audio-visual targets. However, in a condition were “Behavior and Basal Ganglia” research unit (EA 4712), University of Rennes 1, we use complex stimuli such as optical flow in the presence of audio signals mod- Centre Eugene Marquis, Department of Oncology, Rennes, France ulated in amplitude little is known about the processing of multisensory integra- Basal ganglia (BG) are well known for their role in the control of movement. tion in this structure. Furthermore, since sensory deficits extending beyond the However, the concept of control remains vague when considering the non-motor auditory domain constitute hallmark signs in the aging population, we sought to functions of the BG and their role in planification and preparation of action,in gain some insight on whether the time course of complex audiovisual processing the broadest view of this term. in the SC changes with age per se. Fortunately, in addition to its therapeutic effect, deep brain stimulation (DBS) of Materials /Methods: Extracellular single-unit recordings were conducted in the the BG in humans offers the opportunity to directly modulate or directly assess superficial and intermediate layers of the SC of anaesthetized Sprague-Dawley electrophysiological activity of the subcortical nuclei in motor and non-motor adult control (6–12 months) and aged rats (20–22 months). Dynamic concentric tasks. As a consequence, a large amount of data on the role of the subthalamic sinusoidal visual stimuli were presented alone or simultaneously with a Gaussian nucleus (STN) has been accumulated for 20 years. This is largely due to the fact sinusoidal noise in free-field in pseudo-random order at different spatial and tem- that the STN is the most commonly implanted nucleus for the treatment of Par- poral frequencies. kinson’s disease (PD) and that this nucleus focuses research for its therapeutic Results: A number of differences were noted: generally, a surprising difference outcomes. Initially exclusively considered for its motor functions, the STN was observed in the types of audiovisual interactions encountered in the elderly appears now to be implicated in non-motor functions such as emotional process- rats. Thus, a substantial majority of neurons of the SC display no audiovisual ing, moral decision and social decision-making, detection and expectation of interaction for spatial frequency (adult: 62%; aged: 92%) as well for temporal fre- reward, complex cognitive activities and motor planning which includes motor quency (adult: 65%; aged: 90%). Bimodal interactions were more important in imagery, motor representation and motor preparation. adult rats (spatial frequency: 38%; temporal frequency: 35%), than aged rats, This accumulation of functions may be epiphenomenal since it is obviously only around one tenth of neurons exhibiting a bimodal audiovisual interaction biased by the recent opportunity to study this nucleus in patients undergoing (spatial frequency: 8%; temporal frequency: 10%). DBS, especially for PD. On the other hand, there is to consider that all these pro- Discussion: These results suggest that multisensory neurons in the SC play an cesses share common features and that this apparent ubiquity reflects a more essential role in the processing of dynamic auditory and visual information pres- restricted but crucial function of the STN. Indeed, a number of lines of evidence ent in a complex environment, and that aging has a deleterious effect on this support a role of the STN in inhibition of automatic or habitual responses, allow- multisensory integration. ing appropriate switch between ongoing competitive programs, especially from Keywords: superior colliculus, multisensory integration, optic flow, spatial fre- automatic to controlled actions. The role of the STN in selective disinhibition/ quency, temporal frequency, audition, vision facilitation has thus been demonstrated through manipulation of STN activity using DBS. In addition, recordings of local field potentials (LFPs) have provided evidence for direct involvement and modulation of neuronal STN activity during response inhibition. It was recently proposed that modulation of power in beta- 18-03 band activity in the STN (or in the circuitry involving the STN) was critical in Functional alteration without loss of small nerve fibers is sufficient to increase skin pressure ulcer development in mice the processing of action selection and response during the presentation of con- a b b c a flicting stimuli. Change in beta-band activity seems to play a prominent role in A Danigo , L Magy , B Funalot , C Demiot FacultedeMedecine et Pharmacie, EA6309 Maintenance myelinique et neuropathies peripheriques, Limoges cedex, such processes but more recently was also reported modulation in other fre- b quency bands, including alpha, theta and gamma bands. Further, as the level of France; FacultedeMedecine et Pharmacie, EA6309 Maintenance myelinique et neuropathies peripheriques, CHU-Centre de reference des neuropathies peripheriques beta activity in the subthalamic area is inversely proportional to the likelihood c that a new voluntary action will need to be processed, the STN seems to modu- rares, Limoges, France; FacultedeMedecine et Pharmacie, EA6309 Maintenance late its activity in respect to the forthcoming action. myelinique et neuropathies peripheriques, Limoges, France These findings are consistent with the proposal that the STN acts as a filter, Objectives: Pressure ulcers (PUs) generate time and cost consuming treatments inhibiting competitive actions (be it primarily motor or cognitive in their nature) and a significant burden in nursing institutions. The nociception impairment in in order to allow achievement of the appropriated action toward its goal. How- diabetic patients is one of the main predisposing factors for developing PUs. Pres- ever, a critical question remains unsolved: is there a direct relation between these sure-induced vasodilatation (PIV), a cutaneous physiological neurovascular (C- findings, described in experimental conditions, and impulsive behaviors observed fibers/endothelium) mechanism, is altered in diabetes and increases the risk of in patients undergoing DBS for PD? PUs. Our objective was to determine the consequence of a specific sensory small nerve neuropathy in mice on pressure-induced ulcer. Materials and methods: Small-fiber neuropathy was induced in mice by injec- 18-01 tion of resiniferatoxin (RTX), a TRPV1 (Transient Receptor Potential Vanilloid 1) agonist expressed in small fibers. Functionnal and histological assessments were Evaluation of attentional reactivity with non contact methods: pupil and carried-on seven days after RTX injection. Sensory neuropathy was characterized blinks eyelid responses to emotional pictures a a b b a by thermal (Hot-plate) and mechanical (Randall-Sellito) nociceptive tests. Densi- PL Martin , F Andersson , J Martineau , N Hernandez UMR U 930, Universite ties of intraepidermal nerve fibers (DIENF) positive for PGP9.5 and of neurons in Francßois Rabelais de Tours, CHRU Bretonneau de Tours, Tours, France; bUMR U ß dorsal root ganglia (DRG) were quantified to determine nerve degeneration. Mor- 930, Universite Francois Rabelais, CHRU Bretonneau, Tours, France phological aspect of unmyelinated fibers was determined in sciatic nerve by elec- Number of disorders with prominent emotional disturbances (depression, anxiety tron microscopy. Calcitonin gene-related peptide (CGRP) and substance P (SP) disorders, bipolar disorders, PTSD and schizophrenia (DSM-IV-TR)), manifest expressions were quantified in DRG and in epidermal nerve fibers. Skin microvas-

cular functions were evaluated by acetylcholin or sodium nitroprusside iontopho- 18-06 resis delivery and PIV was measured by laser doppler flowmetry. Skin pressure Disruption of 5-HT2A receptor/PDZ protein interactions enhances the ulcers were induced by application of magnetic devices on mouse back during effect of fluoxetine: differential effect on touch-induced allodynia and 12 h, which causes local ischemia. PUs development was evaluated during the spontaneous mechanical hypersensitivity in SNL-induced neuropathic 3 days after magnets withdrawal. rats Results: RTX induced a significant thermal and mechanical hypoalgesia associ- A Dupuisa, A Martinb, AM Privatb, M Chalusb, A Eschalierb, P Marinc, ated with SP depletion. There was no degeneration of intraepidermal nerve fibers C Courteixb aInserm U1107/UdA, Clermont-Ferrand Cedex, France; bInserm U in paw skin and unmyelinated nerve fibers in sciatic nerve were undamaged. 1107/UdA, Clermont-Ferrand, France; cCNRS/IGF, Montpellier, France RTX had no effect on skin microvascular reactivity. RTX mice showed a lack of Background and aims: Antidepressants represent one of the reference treat- PIV associated with CGRP depletion in DRG and their sensory afferents in skin. ments for neuropathic pain. Although activation of serotonergic neurons from Stage 2 ulcer areas were significantly larger in RTX mice at days 1, 2 and 3 after the Rostroventral Medulla reduces pain via descending tracts to the spinal cord, ischemic pressure. selective serotonin reuptake inhibitors (SSRIs) which increase the availability of Conclusion: A specific functional alteration without overt morphological 5-HT, fail to relieve neuropathic pain. This lack of efficacy is due, at least in part, changes or loss of small nerve fibers, associated with a decrease of CGRP and SP to 5-HT2A receptor interactions with intracellular PDZ proteins. The aim of this expressions, increases pressure-induced ulcer development. So, we suppose that study was to compare the effect of fluoxetine, a SSRI, on tactile allodynia and small nerve fiber functions depending on CGRP and SP releases protect skin spontaneous pain in spinal nerve ligature (SNL)-induced neuropathic rats, when against ischemic pressure. administered alone or combined with a peptide disrupting the 5-HT2A receptor- Keywords: skin pressure ulcer, small fiber, neuropathy, resiniferatoxin, pressure- PDZ protein interactions. induced vasodilatation, calcitonin gene-related peptide, substance P Methods: Fourteen days after L5 spinal nerve ligature in rats, we evaluated the effect of fluoxetine on evoked and spontaneous pain using von Frey hair (VFH) test and dynamic weight bearing (DWB) by measuring tactile allodynia and the 18-04 postural equilibrium of freely moving neuropathic rats, respectively. Fluoxetine Effects of the combination of Buprenorphine and Clorazepate on administration (10 mg/kg/day i.p., one injection every half-life/12 h) was per- benzodiazepine binding sites in various brain areas of mice after formed during 15 days. Six hours after every sixth fluoxetine injection, TAT-2A impregnation or not by morphine (a peptidyl mimetic of the 5-HT2A receptor PDZ ligand) was intrathecally admin- O Yildirima, ML Boccab, A Benoitc,LLMad, A Coquerele aService de Pharmacologie, istrated (100 ng/rat) and VFH and DWB tests were performed 30 min after. & Inserm 1075 - UCBN ‘COMETE’, Caen cedex, France; bInserm 1075 - UCBN Results: Fluoxetine administration over 3 days (i.e. six injections) had a slight ‘COMETE’, Caen, France; cInserm U 1075 - UCBN ‘COMETE’, Caen, France; antiallodynic effect, which was enhanced by the acute co-administration of the dService de Pharmacologie CHU de Caen, Caen, France; eService de Pharmacologie & TAT-2A peptide whereas the spontaneous pain behavior was not modified by Inserm U1075 - UCBN, & Addictovigilance - CEIP, CAEN cedex, France either the repeated administration of fluoxetine or the co-administration of fluo- Introduction: In France, recent analyses showed opiate substitution drugs are xetine and TAT-2A. A longer treatment duration failed to improve spontaneous used by 143.000 people and more of 70% used high dosage buprenorphine pain behavior. (BPN). Although BPN is globally well tolerated specific studies (i.e. ‘OPPIDUM’) Conclusion: The data presented here indicate that (i) the DWB apparatus may showed persistent abuses and misuses of BPN with various benzodiazepines provide a new approach to assess spontaneous mechanical hypersensitivity in the (BZD). Previously we described changes in binding for the l opiate receptor SNL model of neuropathic pain and (ii) the repeated administration of fluoxetine induced by BZD in rodents. Reciprocally we hypothesized the BZD binding could combined with acute TAT-2A alleviates tactile stimulation-evoked pain but fails be changed by a chronic morphine [Mor] exposure and later eventually changed to improve spontaneous pain behavior suggesting different pathophysiological after addition of BPN or a BZD or their association. mechanisms involved in spontaneous and evoked pain. Material & methods: We tested mice (male, Swiss type; 25–35 g; 5 by group) Keywords: Neuropathy, SSRI, rats, allodynia, spontaneous pain behavior receiving either Mor (20 mg/kg SC) or saline during 15 days and then a short exposure to BPN (1 mg/kg SC) or (clorazepate [CRZ], 4 mg/kg IP) or their association vs. saline. Brain sections (20 lm) were incubated with 3H-flunitrazepam 18-07 and BZD binding characteristics (number of binding sites [Bmax] and affinity Event related potentials elicited by violations of auditory regularities in [KD]) were measured and computed with Beta-Imager (Biopsace lab, Paris). Sta- patients with impaired consciousness a b c a tistical differences were calculated with a two way ANOVA. F Faugeras , JM Achard , L Naccache CHU Limoges, Limoges cedex, France; b c Results: Changes in Bmax and KD were very different depending on whether the CHU Limoges, Limoges, France; ICM Pitie Salpêtriere, Paris, France treatments were ‘acute’ or ‘chronic’. Of course the anatomical differences were Purpose: To investigate the relevance of neurophysiological assessment of disor- also important. In the frontal lobe the acute opiate effects were limited to a signif- ders of consciousness patients to reveal conscious processings. icant (P < 0.05) decrease in KD without effect on Bmax. After chronic Mor expo- Material and methods: Ten control subjects and 49 acute or chronic disorders sure (CME) a significant decrease in Bmax was found with BPN + CRZ. In of consciousness (vegetative, minimally conscious or conscious) patients were amygdala: after CME Bmax significantly decreased with CRZ (P < 0.05) and CRZ enrolled in this study. + BPN (P < 0.01) and KD significantly decreased after Mor, CRZ and CRZ+ BPN Patients were thoroughly evaluated by using a dedicated clinical scale (Coma (P < 0.02). In hippocampus changed were restricted to a Bmax decrease after Recovery Scale-Revised (Kalmar and Giacino Neuropsychol. Rehabil., 2005)). CRZ + BPN in the CME group. Both control subjects and patients (at bedside) were stimulated by a recently Discussion & conclusion: Taken together these results (i) confirmed a crosstalk described auditory rule extraction paradigm while their scalp EEG (electroenceph- between the regulation of BZD and l receptors after Mor exposure and substitu- alographic) activity was recorded to extract ERP(event related potentials) tion treatment (ii) the association of BPN + CRZ increased binding changes espe- responses to both local (three local effects) and global (global effect) violations of cially in areas implicated in craving behavior. auditory regularities embedded in this paradigm (Bekinschtein, Dehaene et al., Keywords: pharmacodependance, opiates, substitution treatment, buprenor- PNAS, 2009). These effects were evaluated at both group and individual levels. phine, benzodiazepine, binding Results: Group analysis: - Local effects: At least, one of the three local effects was significantly present in each group (all P < 0.01). 18-05 Amplitude of the first of these local effects (mismatch negativity) was related to STIM1 is indispensable for Ca2+ signaling and fat preference in mice the level of consciousness. The more conscious the subjects were, the larger the S Abdoul-Azizea, G Dramaneb, S Selvakumara, H Sadouc, P Besnarda, MMN amplitude was. N Khana aPhysiologie de la Nutrition et toxicologie, UMR INSERM U866, Universite Global effect: This effect was only observed in the group of control subjects. de Bourgogne/Agro-Sup, Dijon, France; bLaboratoire de Biochimie, Universitede Individual analysis: - Local effects: Parakou, Parakou, Benin; cLaboratoire de Nutrition, Universite Abdou Moumouni, Mismatch negativity presence was affected by the level of consciousness but was Niamey, Niger present in some patients in each group. The lipid-binding glycoprotein CD36, expressed by circumvallate papillae (CVP) of Global effect: Global effect was only present in conscious patients. This effect was the mouse tongue, has been shown to be implicated in oro-gustatory perception surprisingly presented in two vegetative patients who interestingly regained con- of dietary lipids. We demonstrate that linoleic acid (LA) by activating sPLA , sciousness within a few days. 2 Another interesting expectancy effect reminiscent of the contingent negativity cPLA2 and iPLA2 via CD36, produced arachidonic acid (AA) and lyso-phosphati- dylcholine (Lyso-PC) which triggered Ca2+ influx in CD36-positive taste bud cells variation was isolated; the slope of which was related to the subjects’ status of (TBC), purified from mouse CVP. LA induced the production of Ca2+ influx factor consciousness. (CIF). CIF, AA and Lyso-PC exerted different actions on the opening of store-oper- Discussion: The global effect was only present in conscious patients or in ated Ca2+ (SOC) channels, constituted of Orai proteins and regulated by STIM1, a patients who regained consciousness rapidly. So, it could be a neural signature of sensor of Ca2+ depletion in the endoplasmic reticulum. We used siRNA technol- consciousness despite its low sensitivity. From a theoretical point of view, this ogy and transgenic mice models and observed that CIF and Lyso-PC opened result is of great importance because global effect seems to emerge from a net- Orai1 channels whereas AA-opened Ca2+ channels were composed of Orai1/ work of generators close to the conscious global workspace (Dehaene and Nacc- Orai3. STIM1 was found to regulate LA-induced CIF production and opening of ache, Cognition, 2001). both kinds of Ca2+ channels. Furthermore, Stim1–/– mice lost the spontaneous Keywords: consciousness, ERP, neurophysiology preference for fat, observed in wild type animals. The CD36-positive TBC from Stim1–/– mice also failed to release serotonin into extracellular environment. Our results suggest that fatty acid-induced Ca2+ signaling, regulated by STIM1 via 18-12 CD36, might be implicated in the transmission of gustatory information on die- Subjective and physiological emotional reactivity in normothymic bipolar tary lipids towards afferent nerve fibers. patients Keywords: calcium, taste buds, lipids, CD36 M Lemairea, N Hernandezb, J Martineauc, F Bonnet-Brilhaultd,WEl Haged aCHRU Tours/Facultedemedecine de Tours, Tours, France; bUniversite Francßois Rabelais, Tours, France; cINSERM U930, Tours, France; dCHRU Tours- FacultedeMedecine de Tours -INSERM U930, Tours, France Introduction: The normothymic phase in bipolar disorders is generally considered to be symptom-free, which is controversial. Many authors emphasize that residual symptoms, such as emotional reactivity disturbance, are under-

evaluated. The aim of this study was to compare emotional reactivity between cytes. Using two-electrode voltage-clamp techniques, we measured the EFX effects normothymic bipolar patients and healthy controls. on GABA-evoked currents. Our results showed that EFX potentiation of GABA- Methods: We evaluated emotional reactivity in 26 normothymic bipolar patients evoked currents depends on the composition of extrasynaptic GABAA receptors. and 30 controls, using an emotional induction method based on the viewing of a Alpha 5 containing receptors were less sensitive to EFX than alpha 2 containing set of 36 pictures (12 negative, 12 neutral, 12 positive) extracted from the Inter- receptor. Surprisingly, we observed that alpha 6 containing receptors GABAA national Affective Picture System. We evaluated subjective emotional reactivity were insensitive to EFX. Since extrasynaptic GABAA receptors are insensitive to (valence and arousal) with the Self-Assessment of Manikin and physiological benzodiazepine molecules, our findings suggested an alternative-binding site of reactivity measuring the pupil response with an eye-tracking material. EFX, which may reflect its physiological effects. Results: The dilation of the pupil was significantly lower in normothymic bipolar References: patients during the viewing of positive pictures (P < 0.05). However, no differ- Hamon A., et al. Neuropharmacology (2003) 45 293–303. ence was found between normothymic bipolar patients and controls regarding Farrant M, Nusser Z. Nat. Rev. Neurosci. (2005) 6 215–229. the subjective emotional reactivity. Keywords: GABAA receptors, etifoxine, Xenopus oocyte, two electrode voltage- Discussion: Compared to controls, normothymic bipolar patients had less emo- clamp tional reactivity to positive valence. This result is in coherence with the more frequent negative emotional bias (sadness, anxiety and anger) observed usually in bipolar patients. These preliminary results need to be replicated. 18-10 Keywords: Bipolar Disorder, Emotional Reactivity, Emotions, Pupil, Affective Spatial learning abilities following omega-3 supplementation are related pictures to hippocampal neurogenesis but not to fat distribution in mice D Deplanquea, P Kafaraa, F Augera, R Bordeta aUniversite Lille-Nord de France, FacultedeMedecine, Lille, France 18-08 Introduction: In a preliminary study, we showed that a 6-week duration Can oesophageal motility be modified by deep brain stimulation: result of omega-3 supplementation may improve memory function in mice in parallel to a randomized cross-over study hippocampal neurogenesis (1). Here, we more precisely investigated the effect of G Gourcerola, S Derreya, N Chastana, D Maltêtea,EVerinb, J Webera, omega-3 supplementation on spatial learning abilities in mice and the possible AM Leroia aCHU Rouen, Rouen, France; bCHU Rouen, CHU Rouen, France role of both hippocampal neurogenesis and energetic changes. Introduction/Objectives: While the role of dopaminergic nigrostriatal loop on Methods: Male Swiss mice (20–25 g) received a supplementation with alpha-li- the control of movements, little is known on its impact on gastro-intestinal motil- nolenic acid (ALA, Sigma France, 250 mg/kg, gavage), peanut oil or water (same ity. Deep brain stimulation (DBS) is currently used to treat Parkinson’s disease volumes) during 6 weeks before to be submit to behavioral evaluations using y- (PD) by decreasing sub-thalamic dopaminergic output. Recently, DBS has been maze and Morris water maze. As possible markers of neurogenesis, we analyzed shown to stimulate gut cholinergic contractions through a central dopaminergic the hippocampal volume and hippocampal blood volume variations using Mag- loop (Neuroscience;195:89-). We therefore took the opportunity to investigate netic Resonance Imaging (2) as well as modifications at cellular level according the effect of DBS on esophageal motility in PD patients in an interventional ran- to double-cortin labeling. Moreover, we analyzed the possible impact of fat distri- domized study. bution through the use of Lunar PIXImus densitometer. Aims & Methods: Seventeen patients (age: 62+/À9 yo) with chronic bilateral Results: A 6-week ALA supplementation was associated to an improvement of high frequency of the subthalamic nucleus for at least 6 months were enrolled in learning abilities in mice since all performances in both the y-maze and Morris this study and randomized with stimulator either turned OFF (2 h) then ON water maze were clearly improved. Besides the observation of greater learning (2 h), or turned ON (2 h) and OFF (2 h) thereafter. Esophageal high-resolution abilities in the Morris water maze during the training period, the time spent in manometry was performed at the end of each period, with a 5 min resting period the quadrant that normally contains the platform was about twice higher in sup- followed by ten swallows of 5 mL. plemented animals (ALA 23 Æ 7 s; Peanut oil, 11 Æ 6 s; Water, 13 Æ 3s; Results: No group’s effect (ON-OFF vs. OFF-ON) was observed for the different n = 23–29 in each group; P < 0.01). In parallel, we observed an increase in both parameters analyzed. During the ON period, a significant increase of the distal hippocampal neurogenesis and hippocampal volume. In opposition, body weight contractility index was found (OFF: 1640 Æ 719 vs. ON: and fat distribution were similar whatever the treatment group. 2035 Æ 598 mmHg.cm.s; P = 0.03), whereas no difference in the distal front Conclusion: Chronic omega-3 supplementation improves spatial memory and velocity was noted compared to the OFF period (OFF: 3.3 Æ 0.4 cm/s vs. ON: learning abilities in mice together with an enhancement of hippocampal neuro- 3.1 Æ 0.2 cm/s; P = 0.49). In addition, a decrease of the integrative relaxation genesis. Such an approach may be of interest in Alzheimer disease to preventively pressure of the lower esophageal sphincter was noted for 1 s (OFF: increase cognitive reserve. 7.7 Æ 1.7 mmHg vs. ON: 4.5 Æ 1.4 mmHg; P = 0.05) and 4 s (OFF: References: 11.1 Æ 1.8 mmHg vs. ON: 7.2 Æ 1.8 mmHg; P = 0.05) during the ON period 1. Deplanque D. et al. Long duration omega-3 supplementation may improve spa- compared to the OFF period, while the resting pressure of the lower esophageal tial memory in mice in parallel to an enhancement of hippocampal neurogenesis. remained similar during the two periods (OFF: 22.2 Æ 2.8 mmHg vs. Fund Clin Pharmacol (2012) 26(S1) 20. ON:18.7 Æ 2.5 mmHg; P = 0.27). This resulted in a significant rise of the intra- 2. Pereira A.C., et al. An in vivo correlate of exercise-induced neurogenesis in the bolus pressure (OFF: 4.5 Æ 1.9 mmHg vs. ON: 2.4 Æ 1.3 mmHg; P = 0.03). adult dentate gyrus. PNAS (2007) 104 5638–5643. Last, no difference were observed during the two period for resting or relaxation Keywords: omega-3, hippocampus, neurogenesis, MRI, double-cortine, fat pressure of the upper esophageal sphincter, nor the pharyngeal contraction amplitude and velocity. Conclusion: It is concluded that DBS increases esophageal body contractions 18-11 and enhances the lower esophageal sphincter opening. This suggests therefore Erythropo€ıetin protects newborn rats against sevoflurane-induced that the central nigro-striatal loop is involved in the control of gut motility and neurotoxicity its impairment may participate on the gastro-intestinal symptoms often associated L Pellegrinia, L Vellya, N Bruderb, G Hachec, B Guilletc, P Pisanoc aINSERM with PD. UMR-S1076 - Faculte de Pharmacie/Service Anesthesie Reanimation - CHU timone, Keywords: Maladie de Parkinson noyaux gris centraux Motricite digestive deglu- Marseille, France; bService Anesthesie Reanimation - CHU Timone, Marseille, France; tition cINSERM UMR-S1076 - Faculte de Pharmacie, Marseille, France Introduction: Recent data obtained in newborn animals exposed to anesthetics have raised serious safety concerns regarding current anesthesia practice in 18-09 young children. Indeed, studies in rodents have demonstrated a widespread Differential sensitivity of synaptic and extrasynaptic GABAA receptors to increase in brain apoptosis shortly after exposure to sevoflurane followed by a the anxiolytic compound etifoxine: implication of the subunit long-term neurologic impairment. In this context, we aim to evaluate the protec- composition tive effect of rh-EPO, a potent neuroprotective agent, in rat pups exposed to sevo- C Matteia, A Hamonb, M Verleyec, D Henriona, NC Guerineaua, C Legrosa aCNRS flurane. UMR6214, Inserm U1083, Univ. Angers Faculty of Medicine, Angers, France; Material and Methods: Seventy-five postnatal day 7 rats pups were allocated bUniversite d’Angers, Angers, France; cBiocodex, Service de Pharmacologie, Compiegne, into three groups: EPO (n = 27): exposed to sevoflurane 2 vol% (0.5 CAM) for France 6 h in a mixture air/O2 (60/40) + 5000 UI/kg rh-EPO IP; SEVO (n = 27): Etifoxine (EFX, Stresamâ), 2-ethylamino-6-chloro-4-methyl-4-phenyl-4H-3,1-ben- exposed to sevoflurane + vehicle IP and CONTROL (n = 21) exposed to the mix- zoxazine hydrochloride) is an anxiolytic and anticonvulsant compound, which ture without sevoflurane + vehicle IP (IP: Intra Peritoneal). Three days after potentiates GABAA receptor function through two mechanisms: (i) by direct anesthesia (D10), quantification of apoptosis was performed on brain extract with binding to GABAA receptors and (ii) by directly stimulating the synthesis of neu- TUNEL method and caspase3, NGF and BDNF expression was determined by roactive steroids in the brain. However, the structure-function relationship of the Western blotting. Rats reaching adulthood (D100) were evaluated in terms of direct interaction of etifoxin with GABAA receptors is still not well understood. exploration capacities (object exploration duration) together with spatial and Using recombinant murine GABAA receptors expressed in Xenopus oocytes object learning (Water maze and Novel object test). (Hamon et al., 2003), we previously showed that GABAA receptor beta subunit Results: Postnatal sevoflurane exposure impaired normal behavior in adult rats is crucial for the mode of action of EFX and that the EFX binding site is different by reducing the exploratory capacities during the novel object test and impaired than that of benzodiazepine. The GABA-evoked current potentiation induced by both spatial and objects learning capacities in adult rats (Water Maze; Place trial: EFX has been shown to be more important at receptors containing a beta2 or a Ratio time to find platform 3rd trial/1st trial: 1.1 Æ 0.2 vs. 0.4 Æ 0.1; n = 9, beta3-subunit than at receptors incorporating a beta1-subunit. Here, we exam- SEVO vs. CONTROL; P = 0.01). Rh-EPO reduced sevoflurane-induced behavior ined the potentiating effect of EFX towards synaptic and extrasynaptic GABAA and learning troubles in adult rats (Water Maze; Place trial: Ratio time to find receptors, using Xenopus oocytes. The presence of a gamma or a delta subunit platform 3rd trial/1st trial: 0.3 Æ 0.1 vs. 1.1 Æ 0.2; n = 9, EPO vs. SEVO; guides the receptor localization at GABAergic synapses and recruits these recep- P = 0.01). Three days after anesthesia, rh-EPO prevented sevoflurane-induced tors in phasic or tonic neuronal inhibition (Farrant & Nusser, 2005). In addition, brain apoptosis (5 Æ 3 vs. 35 Æ 6 apoptotic cells/mm2; n = 6, EPO vs. SEVO; synaptic and extrasynaptic receptors exhibit specific subunit composition and P = 0.01) and elevation of caspase three level and significantly increased the pharmacological properties, especially their sensitivity towards anxiolytic mole- brain expression of BDNF and NGF (n = 6, EPO vs. SEVO; P = 0.01). cules. We cloned alpha3, alpha4, alpha5 and alpha6 cDNAs from mice brain Conclusion: A 6-h sevoflurane anesthesia in newborn rats induced significant and co-expressed subunit combinations corresponding to extrasynaptic (alpha4/ long-term cognitive impairment. A single administration of rh-EPO immediately beta3/delta, alpha5/beta3/gamma2 and alpha6/beta3/delta) or synaptic (alpha3/ after postnatal exposure to sevoflurane reduced both early activation of apoptotic beta3/gamma2 and alpha6/beta3/gamma2) GABAA receptors in Xenopus oo- phenomenon and late onset of neurological disorders.

Keywords: newborn, neuroprotection, anesthetic-induced toxicity, erythropoietin recruited. Maximal coupled mitochondrial respiration (State 3, St3) was determined ex vivo by high-resolution respirometry in saponin-permeabilized myofibers obtained from percutaneous biopsies of vastus lateralis. Maximal uncoupled respiration was achieved with the addition of carbonyl cyanide p-tri- SESSION 19: GS BIOLOGIE DE L’EXERCICE fluoromethoxy-phenylhydrazone (FCCP). Mitochondrial respiratory chain complex content and activity were measured in vitro. In vivo maximal phosphorylation capacity under oxidative condition (ATPmax) of vastus lateralis was determined 31 19-01 by P magnetic resonance spectroscopy. Results: Ex vivo St3 mitochondrial respiration rate was significantly correlated Role of cerebral blood flow in physical exercise-induced brain plasticity 2= = H Banoujaafara,b, A Prigent-Tessiera,b, C Mossiata,b, C Mariea,b aINSERM U1093 with in vivo ATPmax (R 0.52, P 0.0008). However, the graphical distribution b of ATPmax values plotted against St3 values clearly indicated a different slope Cognition, Action et Plasticite Sensorimotrice, Dijon F-21078, France; Universitede 2 between the subsets of low and high St3 values: R was .07 (P = 0.50; 9 sub- Bourgogne, Faculte de Pharmacie, Dijon F-21079, France, Dijon, France = Aim: Physical exercise (EX) is the most powerful intervention to increase cogni- jects) and .58 (P 0.02; 9 subjects) respectively. ATPmax was higher in the tive abilities. While increased production of brain-derived neurotrophic factor high-St3 group. The mitochondrial respiratory complex content and activity did (BDNF) by the brain is largely involved in this positive effect of EX, the way by not differ between these two groups. The uncoupled oxidative flux under FCCP was significantly lower in the lowest St3 group and it was very positively corre- which EX impacts cerebral BDNF metabolism remains speculative. Cerebral blood 2 = = flow increases during EX. In addition, several animal and human studies provide lated with ATPmax (R 0.61, P 0.01) only in the high-St3 group. While the arguments for a link between cognitive abilities and brain perfusion. Therefore, respiratory control ratio did not differ between the two groups, this last result we investigated the contribution of cerebral hemodynamics in EX-induced BDNF indicated that FCCP could interact with the phospholipid bilayer only in the overproduction. high-St3 group. Material and methods: Experiments were performed in adult male Wistar rats. Discussion: Our study suggested that along the continuum of muscle energy EX was induced by a 30 min-long daily walking (30 cm/s) on a horizontal tread- output, alteration of the electrons flux through the mitochondrial inner mem- mill for 7 consecutive days. BDNF levels were measured in the cortex 24 h after brane could be a mechanism associated to the impairment of the mitochondrial the last exercise boot using Western Blotting analysis. Definitive occlusion of both coupling process among older persons. common carotid arteries (2VO model) was used to prevent EX-induced cerebral Reference: blood flow elevation. Experiments were performed in exercised rats with previous 1. Coen P.M., et al. Skeletal muscle mitochondrial energetics are associated with 2VO (n = 6) or sham procedure (n = 6). EX was started 3 days after 2VO. Seden- maximal aerobic capacity and walking speed in older adults. J Gerontol A Biol Sci tary rats with (n = 6) or without (n = 6) 2VO were run in parallel. Med Sci (2012); Epub ahead of print. PMID: 23051977 Results: 2VO alone induced severe animal death within the first two days. Sur- Keywords: Aging, Muscle, Mitochondria, Energetic viving 2VO-rats had no difficulty to be exercised and showed a 16% reduction of BDNF levels. While BDNF production was increased by a factor2.5 inrats without interruption of the carotid flow, the increase felt to 20% in rats with 2VO. 19-04 Discussion: Interruption of carotid blood flow dramatically blunted EX-induced Efficiency of skeletal muscle mitochondrial coupling is a critical factor cerebral BDNF overproduction. This data suggests cerebral hemodynamics as a for maximal exercise capacity and oxygen uptake in rats possible mechanistic link underlying the association between EX and brain plas- AI Schlagowskia, F Singha, AL Charlesa, F Piquardb, B Genya, J Zolla aUniversite ticity. de Strasbourg, EA 3072 “Mitochondrie, stress oxydant et protection musculaire”, Keywords: exercise, BDNF, cerebral blood flow Strasbourg, France; bUniversite de Strasbourg, EA 3072 “Mitochondrie, stress oxydant et protection musculaire”, Srasbourg, France The role of mitochondrial oxidative phosphorylation (OXPHOS) efficiency in the 19-02 determination of maximal oxygen uptake (O2max) and maximal exercise capacity = Chronic AMPK activation improved mitochondrial function and is still unknown. Rats were divided in a control group (CTL, n 8) and a group treated with 2-4-dinitrophenol, a mitochondrial uncoupler, for 28 days (DNP, endurance capacity of aged myostatin deficient mice = M Paulya, B Chabib, A Bonnieub, S Mateckia, C Ramonatxob aInserm U1046, 30 mg/kg/day in drinking water, n 8). Using a breath-by-breath system with a Montpellier, France; bINRA, UMR866, Montpellier, France mask, we measured gas exchange during an incremental treadmill test. Basal A hypermuscular phenotype has been reported by naturally occurring mutations (V0) and maximal (Vmax) mitochondrial respiration of skinned fibres were mea- or by genetic manipulation of the myostatin (mstn) gene in many animals species sured in the gastrocnemius muscle, and the effects of DNP were tested in L6 myoblasts. The DNP group presented significantly lower body mass (P < 0.05) and in a human child. Mstn inhibition represents a great hope for the treatment < of muscle-wasting diseases but inversely a potential misuse of gene therapy for and a higher resting O2 (P 0.005), showing an efficient mitochondrial uncou- the purposes of enhancing athletic performance. Beyond hypertrophy, mstn-defi- pling effect of DNP. The incremental treadmill test showed that maximal running speed and running economy (P < 0.01) were lower, whereas O2max was higher cient muscles show specific contractile and metabolic features. Several studies < have reported that loss of mstn expression is associated with a decrease in mito- in DNP-treated rats (P 0.05). In gastrocnemius skinned fibres, V0 was higher (P < 0.01), whereas the acceptor control ratio (ACR, Vmax/V0) was significantly chondrial content that reduces muscle oxidative capacity. AMP-activated protein < kinase (AMPK), a major sensor of cellular energy status, can stimulate mitochon- lower in DNP-treated animals (P 0.05), showing a reduction of the OXPHOS drial biogenesis through the PGC-1a (PPARgamma coactivator-1alpha) pathway, efficiency. A positive correlation was found between ACR and running speed (r = 0.78; P < 0.001). In condition of high ADP concentration, DNP reduced the as well as through catabolic pathways that fuel energy production.The aim of < the present study was to examine the mitochondrial metabolism and the aerobic mitochondrial capacity of myoblasts to produce ATP (P 0.01). These results running performance of myostatin-deficient aged mice (KO), a hypermuscular show that skeletal muscle mitochondrial coupling is a critical factor for maximal genetic model treated or not with an AMPK activator, the AICAR (5-aminoimi- exercise capacities independently of oxygen transport system, and emphasize the dazole-4-carboxamide). Twenty-month old male mstn KO and wild-type (WT) importance to study the qualitative aspect of the mitochondrial function and not mice (n = 17–19/genotype) were randomly divided in two groups comprising only the amount of mitochondria. vehicule-treated (WT+ placebo (PL) and KO+ PL) and AICAR-treated (WT+AI- Keywords: 2-4 Dinitrophenol; mitochondrial uncoupling; exhaustive exercise; CAR (AI) and KO+AI) (n = 8–10/group) animals. During four weeks, mice were maximal running speed; gas exchanges; VO2 max treated with either vehicle (0.9% NaCl) or AICAR at a daily dose of 500 mg/kg body weight (5/7 days per week). Before treatment, MstnÀ/À mice showed significant reduction of maximal running 19-05 speed, limit endurance time and limit endurance distance compared to Mstn+/+ French Olympic medallist longevity: a follow up since the first Olympiad mice. Four weeks AICAR treatment induced no significant improvement in aero- J Antero-Jacquemina, A Latoucheb, M Taffletc, F Dord, A Marcd, LA Marquetd, bic exercise parameters in mice, but reduced differences in endurance capacity PLeVane, G Berthelota, A Calmate, G Reyf, JF Toussainta aInstitut de Recherche between MstnÀ/À and Mstn+/+mice. AICAR treatment did not improve mitochon- biomedicale et d’Epidemiologie du Sport IRMES; Universite Paris Descartes, PRES drial content in aged KO mice but improve the activities of complexes of mito- Sorbonne Paris Cite, Paris, France; bConservatoire National des Arts et Metiers CNAM, chondrial respiration chain (complex I-II-II+III-COX) in aged myostatin deficient Paris, France; cINSERM, U970, Paris Cardiovascular Research Center PARCC, mice. Universite Paris Descartes, Sorbonne Paris Cite, UMR-S970, Paris, France; dInstitut In summary, AICAR treatment managed to restore oxidative metabolism initially de Recherche biomedicale et d’Epidemiologie du Sport IRMES, Paris, France; eComite altered by lack of myostatin. This study suggests the importance to improve skel- National Olympique et Sportif Francßais CNOSF, Paris; f INSERM, CepiDc, Kremlin- etal muscle oxidative quality in mstn KO mice. Bicêtre, France Keywords: GDF-8, endurance, maximal running speed, AMPK-activating com- The benefits of regular and moderate physical activity on longevity are well pound, aging, mitochondria known in the general population. Outliers with intense physical activity are less well described and no study has focused on French Olympians. It is not known if vigorous practice confers additional benefits or higher risks in elite athletes. Since 19-03 the first modern Olympic Games (OG) in 1896, training and physical intensity Skeletal muscle energy output among older adults could rely on levels have constantly increased and even reinforced among medallists. electrons flux through the mitochondrial inner membrane The purpose of this study is to compare the mortality of male French Olympic F Pillarda, C Leeuwenburghb, S Wohlgemuthc, D Riviered aExercise Physiology medallists from summer or winter games to the male civilian French population. Department, Purpan Medical Faculty, Toulouse Cedex 9, Toulouse, France; bInstitute Biographic data have been collected for all athletes that have got at least one on Aging, Division of Biology, University of FLORIDA, College of Medicine, Gainesville medal in the OG since 1896 up to 2008; 15% were excluded because of missing - FL 32611; cDepartment of Animal Sciences, University of Florida, College of data. Data of civilian population comes from French life tables. Standardised Mor- Agriculture and Life Science, Gainesville - FL 32611; dExercise Physiology tality Ratio (SMR) was used to compare the mortality of both cohort; confidence Department, Purpan Medical Faculty, 31059 Toulouse Cedex 9, Toulouse, France intervals were calculated by Byar’s method. Introduction: Lower mitochondrial capacity and mitochondrial efficiency were The Olympians cohort was composed of 854 French Olympians medallist, 457 previously reported to be associated with slower walking speed in older adults1. (53.5%) have deceased until the endpoint of this study. The average age at death is 70.5 Æ 16.0 years. Compared with French civilian population, the Olympians This study examined how the ATP synthesis capacity was linked with the mito- = – chondrial oxidative capacity in older adults. cohort present the following SMR results: 0.76 [CI95% (0.69 0.83)] for the Æ whole cohort (1896–2008); 0.78 [CI95% = (0.71–0.87)] related with Olympians Methods: Eighteen older adults (mean age: 77 years 4.6; 10 men and eight – = women) with a fairly broad range of aerobic fitness and walking function were from the OG before Second World War (1896 1936) and 0.65 [CI95% (0.52 a

0.82)] for Olympians from the OG after Second World War (1948–2008). patients who are recreationally active 1–2 times/week (control group) were eval- Age-specific SMR (20–39 years; 40–59 years; 60–79 years and 80+ years) for uated. To determine VO2peak an incremental cardiopulmonary exercise test the whole Olympians cohort were respectively: 0.59 [CI95% = (0.40–0.85)]; 0.63 (CPET) with expired gaz analysis was performed. Six-minutes walking test [CI95% = (0.50–0.79)]; 0.76 [CI95% = (0.67–0.88)] and 0.90 [CI95% = (0.76– (6MWT) was also performed as a measure of physical function. 1.10)]. Results: There was no significant difference in VO2peak between cancer patients This study demonstrates for the first time a lower mortality rate among French before adjuvant therapy (age, 49 Æ 11 years) vs. control group (age, Olympians in comparison with the general population. This lower mortality could 48 Æ 9 years): 23.3 mL/kg/min and 24.6 mL/kg/min respectively (P = 0.43). In be the result of several factors such as (i) maintained long-term vigorous activity; addition, 6MWT distance was not reduced in cancer patients compared with con- (ii) genetic predispositions; (iii) a more favourable environment; (iv) a healthy trol group: 527 m and 533 m respectively (P = 0.73). There was no side effect lifestyle after sports career. attributable to the CPET. Keywords: Longevity, mortality, elite, athletes Conclusions: In breast cancer patients before adjuvant chemotherapy, CPET is feasible and not associated with significant differences in VO2peak than in healthy control group. Functional capacity assessed by VO2peak may be an original physi- 19-06 ological baseline indicator, both for establishing the training exercise program The importance of physical activity in the balance of type 2 diabetes and for evaluating the impact of chemotherapy on aerobic capacity. A French D Lakhdara aUniversite de Mascara, Mascara randomized controlled trial (adapted home-based walking training program dur- – Objectives: to examine the impact of the physical activity on certain variables ing chemotherapy set at 50 60% of baseline VO2peak) is currently being con- such as fasting serum glucose (FSG), glycated hemoglobin (HbA1c), total choles- ducted. terol (TC), triglycerides (TGs), high density lipoproteins (HDL-C), and low density Keywords: cardiopulmonary exercise test, peak oxygen consumption, breast can- lipoprotein (LDL-C), in obese women and men with type 2 diabetes. cer, adjuvant therapy, physical function, exercise training Methods: we’ve realized the study in Mehor Mehieddine Diabetology center, Mascara, Algeria on 28 patients women (BMI = 29.50 Æ 5.56 kg/m2), and 22 patients men; aged 51.11 Æ 10.52, the patient followed no diet, on medication, 19-09 and presenting no degenerative complications. We carried out the study over two Effect of rehabilitation program on GH/IGF-1/IGF-BP3 axis and periods: before physical activity program, and after the physical activity program, Testosterone/Cortisol ratio in patients with COPD and in healthy which consist on walking 30 mn/a day twice a week, during seven months. subjects Results: Comparing the results before and after physical activities, concerning W Mkachera, Y Trabelsib, Z Tabkab alaboratoire de physiologie et d’exploration the physical variables we observed a significant decrease in the weight (À5.62%, fonctionnelle- faculte de medecine Sousse, chebba, Tunisia; blaboratoire de physiologie et P 0.001), and the BMI (À5.25%, P 0.001), also for the biological vari- d’exploration fonctionnelle- faculte de medecine Sousse, Sousse, Tunisia ables. We observed significant decrease in FSG (À27.55%, P 0.01), Glycemia Background: Skeletal muscle wasting commonly occurs in patients with chronic post effort (À35%, P 0.01), Glycemia Post Lunch (À37.60%, P 0.01), Gly- obstructive pulmonary disease (COPD) and has been associated with the presence cemia Post Lunch (À37.60%, P 0.01), in HbA1c (À9.41%, P 0.01), TC of systemic inflammation and endocrinological disturbance. The aim of this study (À8.74%, P 0.01), TGs (À11.72%, P 0.001), LDL-C (À12.50%, is to analyze the effect of rehabilitation program on the balance of anabolic vs. P 0.001), the rates of TC with HDL-C increased of (À9.40%, P 0.01), but catabolic hormone in patients with COPD and in healthy subjects. for the HDL-C we see a significant increases of (2%, P 0.01). Methods: Seventeen patients with COPD and sixteen age-matched healthy sub- Discussion: These findings show a beneficial of physical activity program on the jects’ underwent exercise training 3 days/week for 8 weeks. Before and after the balance of glycemia and profiles on lipids, associated to an increased energetic de- training program the concentration of growth hormone (GH), Insulin like Growth pense. Factor-1 (IGF-1), Insulin-like Growth Factor-Binding Protein 3 (IGF-BP3), testos- Keywords: physical activity, diabetes millitus, VO2 Max, HbA1c, lipidic profile terone and cortisol in serum were determined. The exercise measurements included a 6-min Walking Test (6MWT). Results: After 8 weeks, there was no significant change in lung function in 19-07 patients with COPD and healthy subjects. There is a significant improvement in Six-minute walking test predicts maximal fat oxidation in obese children the 6-min Walking distance (6MWD) in both groups (P < 0.01). Dyspnea and E Maknia, M Elloumib, Z Tabkab aLaboratoire de Physiologie et des exploration heart rate at rest and at the peak of 6-min Walking Test (6MWT) decreased sig- fonctionnelle, Sousse, Tunisie; bLaboratoire de Physiologie, FacultedeMedecine de nificantly after training program (P < 0.01). Growth hormone, Insulin like Sousse, Sousse, Tunisie Growth Factor-1 and Insulin-like Growth Factor-Binding Protein 3 increased sig- Background: Obesity is associated with reduced exercise maximal fat oxidation nificantly after rehabilitation training (P < 0.01). The rehabilitation program rate (FATmax), which is generally assessed by cardiopulmonary cycling test. The improves the testosterone / cortisol ratio (T/C ratio) in both groups. 6-min walking test (6MWT) presents an alternative method in patients. OBJEC- Conclusion: Pulmonary rehabilitation induces an improvement of the anabolic TIVE: The aim of this study was to establish a practical reference equation facili- process and reduces protein destruction by the modifications in endocrinological tating the prediction of FATmax from the 6MWT in obese children of both factors regulating skeletal muscle in patients with COPD. genders. Keywords: Chronic obstructive pulmonary disease, GH/IGF-1/IGF-BP3 axis, Tes- Design: This study is a cross-sectional study using mixed linear and multiple tosterone/Cortisol ratio, pulmonary rehabilitation regression models. Methods: Anthropometric measurements were recorded and submaximal cycling test and 6MWT conducted for 131 school-aged obese children, 68 boys and 63 19-10 girls. A multiple regression analysis for FATmax, including 6-min walking dis- Performance and morphology in Track and Field tance (6MWD), anthropometric and cardiac parameters as the dependent vari- AMarca, A Sedeaudb, J Schipmana, JF Toussaintc aIRMES, Paris, France; bIRMES- ables, was performed for the two genders separately. Paris descartes, Paris, France; cIRMES-Paris descartes-APHP, Paris, France Results: Mean 6MWD and FATmax were 564.9 Æ 53.7 m and Introduction: Time tends to increase natural variance ie, phenotypic variability 126.5 Æ 12.1 mg/min for boys and 506.7 Æ 55.0 m and 120.7 Æ 10.0 mg/ depending on environmental fluctuations. Morphological traits, weight and min for girls, respectively. The 6MWD, body mass index, Z-score, fat-free mass, height, follow such a law. Sport performance is multi-factorial and encompasses waist and hip circumferences (WC and HC), rest heart rate, and systolic and dia- many physiological and psychological skills, where morphological parameters stolic blood pressures were highly correlated with FATmax for both genders. play a key role. Professionalization strengthened such impacts through morpho- There was a significant correlation between 6MWD and FATmax in both boys type selection. We study here how running distances provide a physiological and girls (r2=0.88 and r2 =0.81, P < 0.001, respectively). Stepwise regression landscape with specific morphological constraints for each race. analyses revealed that the combinations of 6MWD with HC for boys and 6MWD Methods: Body weight, height and BMI were collected for the international ath- with WC for girls improved the predictability of the model (R2=0.81 for boys and letes top100 on seven Track and Field’s events: 100, 400, 800, 1500, 3000, R2=0.72 for girls; P < 0.001). 10 000 m and marathon for the 1996–2011 seasons. This represents a total of Conclusion: In obese children, the 6MWT can be used to predict FATmax when 11 200 athletes or 1600 performers per events. For each event, data were orga- formal test of exercise capacity and gas exchange analysis are unavailable or nized by speed deciles. impractical. It is therefore possible to prescript targeted exercises at FATmax, Results: A gradual hierarchy between running speed, weight, height and BMI is without performing indirect calorimetry, just from a field test. shown for each event decile. Mean weight increased from 57.8 Æ 5.1 kg for Keywords: childhood obesity, cardiopulmonary exercise test, 6MWT, FATmax, marathon’s to 74.8 Æ 6.4 kg for the 100 m runners. This significant trend is prediction equation also present for height and BMI. Furthermore, a gradient of performance is also observed among decile for each event: on long distances, athletes from the first deciles are always lighter and 19-08 smaller than their counterparts of the lowest deciles. On the opposite, sprinters Cardiopulmonary function in breast cancer patients before adjuvant from the fastest deciles are heavier and taller than sprinters from the lowest chemotherapy decile. T Cornettea, MT Antoninia, F Lemaireb,SLeobonb, N Tubianaa,F Conclusion: Optimal morphologies provide strong hierarchical gradients among Vincentc aUniversite de Limoges - EA3842, Limoges; bCHU de Limoges, Limoges, Track and Field’s athletes. France; cUniversite de Limoges - EA3842 - Chaire de Pneumologie Experimentale, Keywords: Morphology, performance, Track and field, anthropometry Limoges, France Background: Research and clinical interest in the therapeutic effectiveness of exercise in breast cancer after surgery has greatly increased. Use of cardiopulmo- 19-11 nary exercise test (CPET) for determination of peak oxygen consumption Sport performances at the Olympics in London 2012, analysis of (VO2peak) provides the gold standard assessment of aerobic capacity. Measure- trajectories a b c b b b ment of VO2peak presents an original approach in clinical oncology research to G Berthelot , A Marck , M Torres , A Marc , J Schipman , A Sedeaud , screening and prescribing exercise interventions for patients with breast can- JF Toussaintb aINSEP/IRMES, Paris, France; bIRMES, Paris, France; cIRMES, Paris, cer.This study evaluate cardiopulmonary function as measured by VO2peak in France breast cancer patients before adjuvant chemotherapy. The development of the best sports performance world has been updated since Methods: Twenty-two patients with breast cancer before adjuvant therapy for the last publication of the article in 2010. We note that the stagnation in the nonmetastatic disease (predominantly stage II) and seventeen non-malignant 90 s in Track and field continues, except for a few rare events where exceptional

athletes still shine. We will also make a detailed analysis of the London Olympics 20-02 this year to see if the trends, especially world records, will be identical to those TNF inhibitor exposure and cancer? Data of case/non case study in obtained in Beijing in 2008. We then found only five new world records in Track French PharmacoVigilance Database and field, with a significant decline in the average performance, and a huge L Salibaa, M Aboutaama, V Rousseaua, L Chebanea, N Petitpainb, B Baldinc, increase of the records in swimming, thanks to the introduction of new swim- P Gilletb, JL Montastruca, H Bagheria aLaboratoire de Pharmacologie Medicale et suits. In the absence of technological innovation, this development of records can Clinique, Centre Midi-Pyrenees de PharmacoVigilance, de Pharmacoepidemiologie, et only be reduced to a much narrower portion. d’Information sur le Medicament, Equipe de Pharmacoepidemiologie, INSERM U1027, Another point of interest that arose at the Olympic games was the publication of CHU de Toulouse, FacultedeMedecine, Toulouse, France; bService de Pharmacologie, a press article in Nature, where the performance of Ye Shiwen was described as Hôpital central, CHU de Nancy, Nancy, France; cCentre Regional de ‘anonymous’. We here focus on the trajectory of young athletes and try to model Pharmacovigilance, Centre Hospitalier Universitaire de Nice, Nice, France the ‘standard’ path or trajectory an athlete would naturally follow. We do under- Background: Tumor Necrosis Factor (TNF) inhibitors can increase lymphoma line that this approach has to be considered as a complementary methodology to risk in patients who have been exposed to these drugs. However, studies consid- traditional biological approaches. ering a potential increase in risk of solid cancers remain inconclusive. The aim of Keywords: Physiology, career, atypicity the present study was to investigate the association between anti-TNF exposure and reporting of cancers using the French Pharmacovigilance database (FPVDB). Methods: All cases with a criteria of ‘seriousness’ registered in FPVDB were 19-12 analysed retrospectively from January 2000 to October 2010. The case/non-case Effects of metabolic rehabilitation program with moderate intensity on method was used to measure the association between occurrence of cancer and weight loss in women exposure to anti-TNF vs. classic immunosuppressant drugs. Cases were defined as MR. Guedjatia, F Achia, D Kadria, H Gacema aService de Physiologie clinique - reports corresponding to the ADR of interest (i.e. cancer) and non-cases were all Centre Hospitalier Universitaire Benflis Touhami, Batna reports of ADRs other than that being studied. Exposure was considered as the Introduction: Obesity is one of the factors of metabolic and cardiovascular dis- presence in a report of the drug interest (i.e. TNF-inhibitor). We applied the case/ eases the most incriminated. The management of this factor raises the interven- non-case method, comparing cases of cancer to other ‘serious’ cases of ADRs tion of the elements that can lead to the onset of obesity, including diet and reported in the database regarding the exposure to anti-TNF in comparison to physical inactivity. other immunosuppressants drugs. The method allowed us to calculate ADR Purpose: An individualized rehabilitation protocol which allows a reduction in Reporting Odds Ratio (ROR) or adjusted ROR (aROR) on risk factors of cancer weight, more precisely a loss of body fat. and its confidence interval. Method: Seveenteen women were selected for the rehabilitation protocol. They Results: During this period, a total of 3315 cases were registered in the FPVD were prescribed a low calorie diet. Cardio respiratory fitness were measured for corresponding to 2035 cases for exposure to at least one anti-TNF and 1280 the identification of training areas and custom between 40% and 50% of oxygen cases for other immunosuppressant drugs. Sex ratio was 0.55 and mean age was consumption (VO2max), the rehabilitation protocol includes three sessions of 1 h 52 years. The mean number of drug taken per patient was 4. Indications were weekly for a month. Weight control was done through bioelectrical impedance rheumatoid arthritis (53%), Crohn’s disease (17%), ankylosing spondylitis (11%) analysis through 6 electrodes at the beginning and the end of the protocol. or psoriasis (7%). A total of 368 (11% of total) reports of cancer was identified Results: Of 17 women selected a mean age of 41 years, 15 responded positively with 305 cases for anti-TNF (15%) and 63 cases (5%) for other immunosuppres- after a month of training. Weight loss is estimated to average 3 kg, Body Mass sant drugs. 88 lymphomas (70 for TNF-inhibitor), and 57 skin cancers (49 for Index (BMI) decreased by 1.3 and total fat mass decreased by 2.3 kg. TNF-inhibitor) were reported. The rate of cancer reporting was significantly Discussion: The weight loss in obese subjects is problematic monitoring; integra- higher for anti-TNF [ROR = 3.41 (2.57–4.51), P < 0.0001]. tion into a customized rehabilitation protocol can help solve this problem. Using Discussion: The rate of cancer reports was significantly higher in patients a readaptaion in moderate intensity of maximal oxygen up take specialy at 40 % exposed to TNF-inhibitors in comparison to patients treated with other immuno- until 50 % of this parameter can help to solve one of the factors of the obesity. suppressant drugs. Haematological and skin cancers were the most frequent types Keywords: rehabilitation- VO2max- reduced weight- total body fat of cancer reported. Keywords: TNF inhibitors, cancer, ADR

SESSION 20: A3P COMMUNICATIONS LIBRES 20-03 Fibrates and risk of cancer for tissues highly expressing PPAR alpha: a nested case control study from the Echantillon Generaliste des 20-01 Beneficiaires F Salvoa, F Bazina, B Begauda, A Parienteb aInserm U657, Bordeaux, France; Reactions to docetaxel: hypersensitivity or injection-related? b G Drabliera, J Hureauxb, D Bourneau-Martina, P Quilleta, A Jameta, L Lagarcea, Inserm U657, Bordeaux, France P Laine-Cessaca aCRPV CHU ANGERS, Angers, France; bPNEUMOLOGIE CHU Objective: To the association between fibrates use and risk of cancer for tissues ANGERS, Angers, France highly expressing PPAR alpha. The aim of this study was to try and determine whether reactions to docetaxel Methods: A nested case-control study was conducted in subjects aged 45 and during its infusion might be due to hypersensitivity or be injection-related. over identified in the Echantillon Generaliste des Beneficiaires (EGB) as of 2004/ The French national pharmacovigilance database was interrogated using the SOC 01/01. Eligible subjects had to have no recorded diagnosis of cancer before ‘immune disorder’ and docetaxel. 2005/01/01. For the main analysis, cases were defined as subjects with incident As at 15 March 2012, 105 cases had been notified. In 34.3% of them, the reac- diagnosis of melanoma, non-melanoma skin cancer, thyroid, pancreas, kidney, or bladder as identified from ICD-10 diagnosis information associated with recorded tion appeared during first cycle of chemotherapy, and in 27.6% during the second cycle. The time to onset was less than 5 min in 62.9% of patients. The incident Affections de Longue Duree (ALD). All incident cases identified between reactions were serious in 30% of cases (including one death). After stopping the 2005/01/01 and 2011/12/31 were eligible. The secondary analysis also consid- infusion, the symptoms regressed within 12 h in 79% of patients, and symptom- ered hospitalisation diagnosis data (PMSI). In both analyses, up to 10 random atic therapy was administered in 82.8% of patients. The cycles were pursued in controls, matched for age, gender, length of follow-up were identified, and comor- 33 patients (31.4%) amongst whom 21.2% were treated at a lower flow bidities. The index date was the date of first recording of cancer in the EGB data- rate, and 87.9% of the patients whose treatment was pursued tolerated it well. base for cases; for controls, it was the date of selection. Use of fibrates was In 24.8% of patients, docetaxel was reintroduced with enhanced premedication assessed using reimbursement data and classified according to the cumulated or under a desensitisation protocol. Only eight reactions appeared to result from number of Delivered Daily Doses reimbursed between 2004/01/01 and day 180 IgE-mediated hypersensitivity amongst whom one positive prick test. before index date. Exposure to drugs considered as potential confounders (statins, In many of these cases, insufficient data were available to diagnose IgE-mediated insulin, oral antidiabetic agents, and aspirin) was assessed similarly. Statistical hypersensitivity, or the data seemed to exclude this hypothesis (negative re-chal- analysis was performed using adjusted multivariate conditional logistic regression lenge, reaction during the first administration). It is possible that the preservative models. used in this medication (Polysorbate 80), and which is also present in food, Results: Thousand two hundred and ninty-two cases and 12830 matched con- might explain the reaction after the first administration. Our findings suggest a trols were included in the main analysis. Mean age of subjects was 70.2 (et different pathophysiological mechanism for intolerance. In the literature (1,2), 10.8), 59.1% were male. Exposure to fibrates was found in 18% of cases and several mechanisms have been described (anaphylactic IgE-mediated reaction, 15.5% of controls. After running adjusted multivariate analysis, exposure to 550 DDD of fibrates was associated with an increased risk for the cancers of anaphylactoid reaction with a non-specific, non-IgE-mediated histamine and tryp- – tase release or with other vasoactive compounds (interleukin, TNF, interferon, interest, all types considered, with an OR of 1,29 (95%CI 1.06 1.57). When con- etc.). sidering associations regarding cancer type, an increased risk was found for non- melanoma skin cancer (OR 1.83, 95%CI 1.13–2.96), and a tendency for kidney To explore this hypothesis, we are currently setting up a study during which lab- – oratory tests for histamine and tryptase will be performed after each intolerance cancer (OR 1.53, 95%CI: 0.96 2.43). In both these, a similar association was reaction. Skin tests will also be performed in the event of histamine or tryptase found for statins. release. Discussion: In this exploratory analysis, use of fibrates was associated with an References: increased risk of cancer for tissue highly expressing PPAR alpha. However, simi- 1. Ardavanis A., et al. Non-allergic nature of docetaxel-induced acute hypersensi- lar associations to statins could suggest a potential indication bias. tivity reactions. Ant-cancer Drugs (2001) 15 581–585. Keywords: fibrates, cancer, pharmacoepidemiology, drug safety 2. Zanotti K.M., et al. Prevention and management of antineoplastic-induced hypersensitivity reactions. Drug safety (2001) 24 767–779. Keywords: docetaxel, hypersensitivity reaction, injection-related reaction 20-04 Pioglitazone and bladder cancer: lost opportunities for risk assessment JL Failliea, P Petita, JL Montastrucb, D Hillaire-Buysa aDepartement de pharmacologie medicale et toxicologie, CHRU Montpellier, Universite Montpellier 1, Montpellier, France; bDepartement de pharmacologie medicale et clinique, Facultede medecine, Toulouse, France Introduction: The risk of bladder cancer associated with pioglitazone has been widely discussed since 1999 and led to its withdrawal in France in 2011.

Methods: We studied the sequence of published scientific evidence regarding this intake of syrup formulation at the same dosage1. The aim of this study is to risk. describe the switch between methadone formulations, using French health reim- Results: We identified three inaccuracies that would have change the safety pro- bursement database in 2010. file of pioglitazone. 1) Before 2000, bladder cancers occurred in male rats treated Methods: This study is based on quasi-exhaustive reimbursement database in with pioglitazone in preclinical studies, mechanistic hypothesis involved pioglitaz- Provence-Alpes-Cote^ d’Azur Corse and Rhone-Alpes^ French areas. Individuals one-induced urolithiasis specific to rats. Authorization labels included these find- from these regions affiliated to the French health reimbursement general system, ings but no specific warning for patient selection and monitoring. Actually, who have had a reimbursement of both formulations of methadone between Jan- specificity to rat appeared to be erroneous in an animal study published in Mars uary 1st and December 31st 2010, have been included. 2011.(1) 2) Pioglitazone has always been presented as a selective agonist for Results: During 2010, 1044 patients (17.5% of methadone’s users in 2010) PPARg. Between 2004 and 2006, all the dual PPARa/g agonists have been dis- have had a reimbursement of both formulation of methadone 76.5% (n = 799) continued due to safety concerns, including bladder tumors. In Mars 2012, study switched from syrup to capsule formulations, including 10.8% (n = 113) who demonstrated that pioglitazone pharmacological profile was comparable to that of went back to syrup. 23.5% (n = 245) switched from capsule to syrup, including the PPARa/g agonists.(2) 3) In 2005, the PROactive trial compared cardiovascu- 14.4% (n = 150) who went back to capsule. 60.8% (n = 635) of patients have lar benefit of pioglitazone to placebo: 14/2605 (0.5%) cases of bladder neoplasms made a single switch, 23.8% (n = 248) have made two switches, 15.4% were reported in the pioglitazone group vs. 6/2633 (0.2%) in the placebo group, (n = 161) have made three switches or more and 18.4% (n = 192) have been difference was not statistically significant (P = 0.069). An overview of the PRO- delivered both formulations simultaneously. active data published in 2009, revealed that one case in the placebo group The sociodemographic profile is the same for these three groups of patients. The showed a benign histology. In 2011, the overall incidence in the placebo group patients who made ‘2 switches or more’ have been delivered more benzodiaze- was recalculated with only five cancers: the risk was shown to be statistically pines than others (69% vs. 60.2% P = 0.036). There is no difference for antide- greater in the pioglitazone group: RR = 2.83 (95% CI: 1.02–7.85, P = 0.040).(3) pressant and analgesic opioids deliveries. Discussion: Whatever the cause of these inaccuracies, opportunities to properly Conclusion: In spite of capsule advantages, 10.8% of patients return to syrup assess the safety profile of pioglitazone have been lost. It is crucial to be particu- formulation and 14.4% have a period with syrup formulation. 18.4% of patients larly vigilant with the safety information provided before and after-marketing. (there who have been delivered both formulations simultaneously) are outside of References: the recommendations. 1. Sato K., et al. Suppressive effects of acid-forming diet against the tumorigenic 1CHLORHYDRATE DE METHADONE CT11413 avis de la commission de la trans- potential of pioglitazone hydrochloride in the urinary bladder of male rats. Toxi- parence de la Haute Autorite de Sante du 29/02/2012. col Appl Pharmacol. (2011) 251 234–244. Keywords: methadone, switch, syrup, capsule, opiate maintenance treatment 2. Hillaire-Buys D., Faillie J.L., Montastruc J.L., Petit P. Stay vigilant: a glitazone (pioglitazone) can hide a glitazar! Eur J Clin Pharmacol. (2012). 3. Hillaire-Buys D., Faillie J.L., Montastruc J.L. Pioglitazone and bladder cancer. 22-03 Lancet (2011) 378 1543–4; author reply 4–5. Risk of major bleeding with dabigatran vs. active controls: a systematic Keywords: Pioglitazone, bladder cancer, risk assessment review and meta-analysis of randomised clinical trials S Antoniazzia, D Berda€ıb, V Contic, P Robinsond, S Radicee, E Clementie, N Mooref, A Parientef, F Salvog aUnit of Clinical Pharmacology, Department of Biomedical and Clinical Sciences, University Hospital “Luigi Sacco” Universitadi SESSION 22: A3P: COMMUNICATIONS LIBRES Milano, Milan, Italy; bCHU Bordeaux, Bordeaux, France; cRegional Centre for Pharmacovigilance, Lombardy Region, Milan, Italy; dUniversite de Bordeaux, INSERM CIC-P 0005, Bordeaux, France; eUnit of Clinical Pharmacology, Department of 22-01 Biomedical and Clinical Sciences, University Hospital “Luigi Sacco”, Universitadi Assessment of triptans abuse in PACA and Corse using a multi-indicator Milano, Milan, Italy; fUniversite de Bordeaux, CHU de Bordeaux, INSERM CIC-P approach 0005, INSERM U657, Bordeaux, France; gUniversite de Bordeaux, INSERM CIC-P D Braunsteina, M Lanteri-Minetb, V Pradela, F Natalic, V Allaria-Lapierrec, 0005, INSERM U657, Bordeaux, France A Donnetd, J Micallefa aCentre d’Evaluation et d’Information sur la Objective: The direct thrombin inhibitor dabigatran could represent an alterna- Pharmacodependance-Addictovigilance PACA Corse, Marseille, France; bDepartement tive drug for atrial fibrillation (AF) and for the prevention or treatment of venous d’Evaluation et Traitement de la Douleur, Pôle Neurosciences Cliniques, CHU Nice, thromboembolism (VTE). As for the other anticoagulants, major bleedings repre- Hôpital de Cimiez, 4 avenue Reine Victoria, Nice, France; cDirection Regionale du sent a major safety issue of this drug. The primary objective of this study was to Service Medical de l’Assurance Maladie PACA-Corse, Marseille, France; dCentre estimate the risk of major bleeding related to dabigatran in randomised clinical d’Evaluation et Traitement de la Douleur, CHU Timone, 264 rue Saint Pierre, trials (RCTs) with respect to active comparators. The secondary objective was to Marseille, France stratify risk according to dabigatran dose and type of active comparator. Introduction: Chronic daily headache induced by triptans overuse is still an Methods: A systematic review and meta-analysis was conducted applying key- important public health problem. The objective of this study was to quantify trip- words related to Dabigatran and Randomised Controlled Trials in Medline, SCOPUS, tans abuse in a French reimbursement database from two French administrative and Cochrane Central Register of Controlled Trials. Active controlled RCTs with regions (PACA and Corse), using a multi-indicator approach. at least 100 patients treated with dabigatran, used at doses approved in clinical Methods: The study population included all people covered by the French Gen- practice for VTE and AF, were included in this meta-analysis. Data were eral Health Insurance System who had at least one dispensation of triptans extracted independently by two investigators and verified by a third one. between May 2010 and December 2011. Delivered quantities were converted Results: Eight trials (34 078 patients) were included in quantitative analysis. into Defined Daily Doses (DDD). Triptans abuse was quantified using three meth- The risk of major bleeding was lower in the dabigatran group than in the control ods. The first method was based on the definition of triptans overuse headache group [risk ratio, RR, 0.87, 95% confidence interval, 95%CI (0.78–0.97)]. Analy- pharmacologic criteria B by International Headache Society (IHS): Triptan intake sis by dose showed a lower risk [RR 0.83, 95%CI (0.73–0.94)] for low doses of on more than 10 days/month (which we translated into 20 DDD/month to dabigatran (100, 150, or 220 mg/day), while for dabigatran 300 mg/day no dif- account for possible double intakes per day) for more than 3 months. The second ference in risk of major bleeding was found [RR 0.92, 95%CI (0.81–1.05)] and method is based on doctor-shopping. This is a patient’s behavior consisting in warfarin was the only active comparator retrieved. Analyses vs. individual con- consulting several physicians to obtain multiple prescriptions of the same drugs. trols, irrespective of dose, showed a lower risk of major bleeding for dabigatran The quantity obtained by doctor-shopping is calculated for each patient, and doc- compared to warfarin [RR 0.87, 95%CI (0.78–0.97)], but no difference vs. enox- tor-shopping indicator is computed for each triptan International Nonproprietary aparin 40 mg/day [RR 1.07, 95%CI (0.72–1.58)]. No evidence of heterogeneity Names (INN). The third method was a mixed classification using principal com- was found. ponent analysis, k-means and hierarchical clustering to identify clusters of sub- Discussion: A lower risk of major bleeding was found for dabigatran compared jects. with active controls, irrespective of dose and control type. However, at the stan- Results: 95540 subjects were included in the study. According to the IHS dard regimen of 300 mg/day that is approved by the EMA and the FDA for AF, method, 2243 subjects (2.3%) were abusers. Their median duration of abuse was risk of major bleeding did not differ with warfarin. 6 month. Among abusers, 1058 subjects (47.2%) were abusers in both 2010 Keywords: anticoagulants, drug safety, quantitative analysis and 2011 study period. The doctor-shopping method highlighted 274 subjects who had a doctor-shopping quantity over 60 DDD. The doctor-shopping indicator was lower than 1% for each INN. The classification method identified five clusters 22-04 of subjects with specific behavior according to total number of DDD, number of Using Hospital medical information system (PMSI) as a tool to improve deliveries, number of prescribers and number of pharmacists delivering triptans. safety signal in pharmacovigilance: about anaphylaxis reactions Conclusion: Our results confirm the need of a multi-indicator approach to assess J Mahea, L Triqueta, N Surerb, G Veyraca, P Jollieta aCHU NANTES - Service de the pharmacoepidemiology of a drug abuse. The behavior called ‘Doctor-shop- Pharmacologie Clinique - Centre Regional de Pharmacovigilance, Nantes cedex; bCHU ping’ seems to be a marginal component of triptans abuse. NANTES - Service d’Information Medicale, NANTES cedex Keywords: pharmacoepidemiology, triptan, abuse Background: Regional Pharmacovigilance Centers are responsible to improve and strengthen French Pharmacovigilance. Conduct research on databases, like hospital medical information system database (PMSI), had been proposed to iden- 22-02 tify adverse drug reactions not spontaneously reported. How methadone’s users switch between syrup and capsule formulations Objective: To assess the interest of PMSI database from university hospital of in two French regions in 2010? Nantes with an example of anaphylaxis reaction reports, to improve safety signal Q Boucheriea, E Fraugera, X Thirionb, M Mallaretc, J Micallefa aCentre in Pharmacovigilance. d’addictovigilance (CEIP) PACA-Corse Marseille, Marseille, France; bCentre associe Methods: PMSI database was retrospectively analyzed over the period January d’addictovigilance (CEIP) PACA-Corse Marseille, Marseille, France; cCentre 2011 to December 2011. ICD-10 codes related to anaphylaxis reactions (T80.5, d’addictovigilance (CEIP) Grenoble, Grenoble, France T88.2, T88.6, T78.2) were used to extract patient’s cases. Each hospitalization Introduction: The methadone syrup is available as opiate maintenance treat- summary was analyzed. We compared our findings with spontaneous ment since many years. More recently, a capsule formulation has been marketed notifications from hospital physicians collected during the same period. This in 2008 with strict supervision for prescription and delivery. Only patients stabi- research was carried out from the French Pharmacovigilance Database using the lized for at least one year with the syrup are eligible for capsule formulation. The following terms: collapse, bronchospasm, urticaria, anaphylactic reaction and an- Health Authorities recommend the first intake of capsule the next day of last gioedema.

Results: Twenty-two files of hospitalized patients were selected among the 44 same intravenous line. Two patients had no re-administration. Among five extracted from the PMSI database during the study period. Meanwhile, 37 spon- patients with re-administration, the adverse effect recurred in two. The 79 chil- taneous notifications were collected by our Pharmacovigilance Center. Twelve dren had a total of 280 infusions; of these 14 were associated with this adverse cases were recorded in both databases: eight were serious anaphylaxis reactions effect [5.0 %; 95%CI (2.8–8.2%)]. Infusion rate and dose were all in accordance grade 3 or 4 according to Ring and Messmer classification (1). Cases from PMSI with the SPC. and cases from spontaneous notifications were similar in terms of gravity regard- Conclusion: These data suggested that venous irritation following intravenous less of anaphylaxis reaction grade. Drugs mainly reported in spontaneous notifi- PPâ infusion remains at an inacceptable level among hospitalized children. Some cations were antibiotics (n = 9) and anesthetic agents (n = 7). Concerning the etiologies (degradation product, acetic acid in excipients) are hypothesed but cur- PMSI cases, antineoplastic agents were largely reported from hematology depart- rently not proved. ment with four cases due to L-asparaginase, leading to treatment discontinua- Keywords: intravenous paracetamol; pediatrics; adverse effect; venous irritation; tion. incidence Discussion: Although analysis time was important, PMSI database allows to collect anaphylaxis reaction with a good recovery: 23% (ratio of selected patients’ files after analyze and exclusion of duplicates on identified total patients’ files). 22-07 This method highlighted 4 anaphylaxis reactions grade 2 or 3 with L-asparagin- Use of the French claims and hospitalisation (EGB) database to assess the ase over the one year study period, while only 3 spontaneous cases were reported prevalence and the incidence of Parkinson’s disease in France to our Pharmacovigilance center since 1997. This study confirms the interest to P Blina, C Dureau-Pournina, A Grolleaua, E Corbillonb, J Jovea, R Lassallea, use PMSI database as a complementary survey method to improve safety signal N Poutignatb, A Foubert-Samierc, C Droz-Perroteaua, N Moored aINSERM CIC-P in Pharmacovigilance. 0005, Universite de Bordeaux, Bordeaux, France; bHaute Autorite de Sante, Saint Reference: Denis La Plaine, France; cINSERM U897, ISPED, Universite de Bordeaux, CHU de 1. Ring J., Messmer K. Incidence and severity of anaphylactoid reactions to col- Bordeaux, Bordeaux, France; dINSERM CIC-P 0005, Universite de Bordeaux, CHU de loid volume substitutes. Lancet (1977) 1:466–469. Bordeaux, Bordeaux, France Keywords: pharmacovigilance, anaphylaxis reaction, using hospital medical Objective: To assess the prevalence and the incidence of the Parkinson’s disease information system in France. Method: The EGB database is a 1/97 permanent random sample of the French healthcare insurance system database linked with the national hospital discharge 22-05 summary database. Data concerning all adults with full insurance coverage for Misuse of short courses of oral glucocorticoids in children: a prospective Parkinson’s disease, or hospitalized with main, related or associated Parkinson’s observational study in a paediatric emergency department disease diagnosis, or with at least three antiparkinsonian drug reimbursements E Bennetta, A Tahirb, V Guigonisb, L Merlec, ML Larochec aEmergency Department, over a one-year period were extracted between 1st January 2004 and 31st University Hospital, Limoges, France; bPaediatric Emergency Department, University December 2010. Among them, two populations were considered, one defined by Hospital, Limoges, France; cCentre of Pharmacovigilance, University Hospital, Limoges, a specific criterion: patients with a medical diagnosis of Parkinson’s disease (full France insurance coverage or hospitalisation); and the other defined by a sensitive crite- Objective: Short courses of oral glucocorticoids appear to be frequently pre- rion: patients with a medical diagnosis of Parkinson’s disease in the database or scribed to children in spite of their iatrogenic potential, the precise mechanisms a drug pattern compatible with this diagnosis (i.e. a second set of at least three of which, notably in terms of necessary dose and duration of exposure, are not antiparkinsonian drug reimbursements over another one-year period and no co- precisely known. The objective of this study was, according to current practice medication with extrapyramidal side effects, as well as no antiparkinsonian drug guidelines, to evaluate the validity of the use of short courses of oral glucocortic- pattern specific of another indication). Prevalence and incidence in France were oids in children and to describe the adverse effects induced. assessed using EGB data, standardized on age and gender distribution of the Method: A prospective observational study was conducted between 7/2010 and year+1 national demographic statistic. 6/2011, including children under the age of 18, who were taking a short course Results: Prevalence of Parkinson’s disease increased from 2.7& in 2005 to of oral glucocorticoids at the time of their admission to a paediatric emergency 3.3& in 2010 with the specific definition, and from 3.8& to 4.6& using the department (Limoges, France). Data for evaluating the validity of the therapeutic sensitive definition. The incidence rate per year was 0.3–0.4& with the specific indications of the short courses of oral glucocorticoids being taken by the chil- population definition and 0.5&–0.6& using the sensitive definition. Estimated dren according to current practice guidelines and the adverse effects observed, prevalence in France was between 180 000 and 255 000 persons in 2010 with were collected using a questionnaire and the patients’ medical file. approximately 22 000–32 000 new patients per year. Results: In total, 100 children aged 4.8 Æ 4.0 years (1 month-17 years) were Discussion: The diagnosis of Parkinson’s disease can rarely be asserted in living included. During the 12 months prior to admission, 56% of children had yet taken patients. Specific diagnostic criteria minimise the number of false positives and an oral corticotherapy. The principal indication of oral glucocorticoids was for sensitive criteria that of false negatives. The Prevalence and the incidence are lower respiratory tract diseases (71%). In fact, 61% of the short courses of oral under-estimated with a specific criterion and over-estimated with a sensitive one. glucocorticoids were prescribed for therapeutic indications not validated by current In this study, both criteria were used to provide an estimate range that should practice guidelines. The most frequent indications were acute cough (42%), acute include the true value. The EGB can be used to assess the prevalence as well as bronchiolitis (16%) and acute bronchitis (8%). Self-medication of the short courses the incidence of a chronic disease such Parkinson’s disease in France, using well- of oral glucocorticoids was encountered in 7% of cases. The incidence of adverse defined specific and sensitive criteria. effects was 38%; these were principally neuro-psychiatric disorders (65%). Keywords: Parkinson’s disease; Prevalence; Incidence; EGB database Conclusion: Systematic use of short courses of oral glucocorticoids cannot be recommended for children in a number of conditions due to insufficient proof concerning their efficiency and relative harmlessness. However, in this study, the 22-08 most frequent misuses concerned conditions for which practice guidelines exist, Severe hyperkalemias associated with the misuse of a fixed drug formally advising against the use of oral glucocorticoids. In light of this informa- combination in hypertensive patients tion it appears essential to emphasize the importance of continuing medical edu- M Andrejaka, J Moragnya, V Brenet-Dufourb, A Cheyrouxa, V Grasa aCentre de cation, the practice of evidence-based medicine and patient education, in order to pharmacovigilance de Picardie - CHU AMIENS, AMIENS; bCentre de recherche clinique reduce unnecessary exposure of children to the potential adverse effects of oral - CHU AMIENS, AMIENS and the French Pharmacovigilance network. glucocorticoids Objective: We will describe cases of hyperkalemia with treatment including a Keywords: oral glucocorticoids, children, misuse, adverse effects, observational drug combination of spironolactone 50 mg and furosemide 20 mg, notified to study French Pharmacovigilance system and included in a national survey. This drug has been largely misused since its authorization in 1992 in France for the treatment of congestive heart failure (Aldalixâ). 22-06 Methods: We analyzed the data concerning the cases notified with kalemia: Venous irritation related to intravenous paracetamol injection 5.5 mM in patients treated by the combination: clinical indication of the treat- LM Scailteuxa, S Picarda, F Aubinb, M Nicollec, P Pladysc, E Polarda aCentre ment, comorbidities, drug associated and outcomes. regional de pharmacovigilance, CHU, Rennes, France; bPharmacie, CHU, Rennes; c Pôle Results: Ninety-three cases of hyperkalemias associated with the use of the fixed de pediatrie, CHU, Rennes, France combination were analyzed. 71 patients (75%) were older than 75 years. Signifi- Purpose: Paracetamol Panpharmaâ (PPâ) obtained a marketing authorization cant comorbidities were diabetes mellitus in 29 cases (31%), previous renal dys- in 2007. The next year, French health professionals reported cases of venous irri- function in 32 cases (32%). Concomitant treatments were NSAIDs taken tation (burning, pain) following intravenous (IV) infusion. Initially, this problem simultaneously in 12 cases (recently in seven cases), ACE in 36 cases, ARA2 in was thought to be related to infusion rate. Then, Panpharma Company has rec- 14 cases and an ACE-ARA2 combination in six cases. Other co-medications with ommended that IV infusion lasted 15 min at least. In 2010, cases of venous irri- a potential to increase kalemia were b-blockers (15), potassium salts (13), other tation were still reported from the pediatric surgery unit of Rennes University aldosterone antagonists spironolactone or eplerenone (3), trimetoprim (2) and Hospital. Infusion rate and dose were in accordance with the Summary of Prod- heparin (1). Ninety-two percents of the cases with data concerning the aim of uct Characteristics (SPC). We set up this study as an attempt to estimate the pro- the treatment were linked to the treatment of high blood pressure. Sinus brady- portion of venous irritation. cardias were described in 17 cases, auriculo-ventricular conduction disturbance Methods: From January 23th to February 06th 2012, all children hospitalized in six cases as well as ECG perturbations potentially consecutive to hyperkalemia. in any of four pediatric units (emergency, neonates, adolescents and surgery) and Two cardiac arrests occurred with 1 fatal outcome. receiving intravenous PPâ infusion, were included. Only peripheral infusions Discussion: Cases of hyperkalemia observed with this combination of spironolac- were included whereas central infusions were excluded. Proportion was then cal- tone and furosemide are associated with a massive off-label use in hypertensive culated as the number of infusion with this adverse effect divided by the total patients (confusion with other combinations of hypo- and hyperkalemic diuretics number of PPâ infusion along with 95% confidence interval. indicated in this setting as a possible explanation). Results: Seventy-nine children with a mean (min-max) age of 6.4 years Furthermore, the use of this combination is not in accordance with the current (20 days-18 years) were included. Seven [8.9 %; 95%CI (3.6–17.4%)] presented recommendations concerning congestive heart failure (use of a large dose of spir- this adverse effect; there were six boys and one girl aged 4–16 year-old. Except onolactone regarding associated dose of furosemide, inability to adjust the relative for one patient, the adverse effect appeared at the 1st or 2nd infusion of PPâ. doses of the two components of the combination according to the follow-up of Five patients had level 2 or 3 analgesics alternately with PPâ infusion and blood potassium). These points led to assess the risk-benefit ratio of the combina- among these, 3 had concomitantly other drugs (antibiotics, NSAID…) on the tion as negative.

The MAH therefore decided recently to cease the commercialization of this fixed The aim of this study was to analyse adverse effects notified to the French phar- combination. macovigilance system and related to nicardipine use whatever the indication. Keywords: hyperkalemia, furosemide, spironolactone, fixed-drug combination, Methods: The analysis concerned case notified to pharmacovigilance regional misuse centers with i.v nicardipine as suspect drug. Results: Two hundred and seventeen cases were analysed. The most frequent adverse reaction reported was infusion site reactions described as veinitis, peri- 22-09 phlebitis, lymphangitis (84) with for some cases the reaction occurring at differ- Intravenous paracetamol: inappropriate indications and doses ents successives sites of infusion. 36 of these cases concerned use of the drug in D Boudesa, P Rodrigueza, E Bondon-Guittonb, JL Montastrucb aServices des preterm labor. Urgences du Centre Hospitalier de Rodez, Rodez, France; bService de Pharmacologie Some adverse effects were directly linked to the vasodilatory effect of the drug: Clinique, Toulouse, France excessive hypotension (13 including six neonates after in-utero expositions), Introduction: Intravenous (IV) paracetamol is indicated in short treatment of symptoms related (headaches, vasomotor flush, palpitations) (8), a few cases of moderate pain or fever, when IV route is clinically justified by the urgency to extrasystoles, sinusal or auriculoventricular bradycardias. treat pain or fever and/or when other routes are not possible. Dose of paraceta- Twenty-three cases of pulmonary edemas have been reported, linked in all cases mol differs according to the weight. In France, some studies have shown that with preterm labor. Outcome was always favourable after discontinuing infusion indication of IV paracetamol is often inappropriate at hospital (58 at 62%) but and in some cases furosemide. At least seven of these cases were linked to multi- we do not know if dose are appropriately administered. We carried out a study in ple pregnancies. Other adverse effects reported were legs edemas, angio-edemas, a French emergency department in 2011–2012 to assess the frequency of various skin reactions including urticarias, acute renal failures, thrombocytope- patients (adults and children) who have an inappropriate prescription of IV par- nias, hepatic reactions (with generally other drugs suspected). Drug interactions acetamol. suspected to be related to nicardipine were reported with tacrolimus in four Methods: This retrospective study was performed in the emergency department cases. of the Public Hospital at Rodez (Aveyron). Adult patients were selected during Discussion: This analysis of the database has shown that adverse effects related non consecutive 3 weeks and randomly chosen (the 49th week in 2011, the 1st to intravenous nicardipine may affect the site of infusion of the drug or be the and 4th in 2012). Children were selected between the 26th May 2011 and the consequence of the vasodilatory action of the drug. Pulmonary edema appears to 17th May 2012. All patients treated with IV paracetamol were included. Those be a side-effect specifically reported in pregnant women with preterm labor and with contra-indication to IV paracetamol or with missing data about temperature may have some favoring factors: increased intravascular volume (especially in or pain were excluded. We checked for all patients if indication and dose of IV multiple pregnancies), concomitant corticosteroids, aggravation of preexisting paracetamol were appropriate or not. alterations in cardiovascular function (pregnancy-related or not). This risk must Results: We included 208 adult patients and 152 children. Indication was inap- be taken into consideration when tocolytic therapy is indicated. propriate in 38.9% of adults and in 21.7% of children. For most of them, oral Keywords: nicardipine, intravenous, preterm labor, adverse effects route was possible. Other patients only suffered from a light pain or had no fever. Dose was inappropriate in 6.3% of adults (overdoses in patients with a weight under 50 kg) and 64.7% of children (mostly overdose). Discussion: This is the first French study assessing the frequency of inappropri- SESSION 23: STP-MODELISATION ate dose of IV paracetamol. New recommendations to adapt the dose to the weight and mistakes between the prescription and the administration could explain inappropriate doses. Emergency context could explain inappropriate indi- 23-01 cations. Oral route, which is safer and less expensive than IV route, should be Fractionation of daily dose increases the predicted risk of severe used such as possible. Finally, dose of IV paracetamol should be prescribed in mg sorafenib-induced Hand-Foot Syndrome (HFS) and ml, especially in children, to prevent overdose. Indications should be E Henina, B Blanchetb, P Boudou-Rouquettec, G Freyerd, F Goldwasserc, respected, especially in adults, to limit the overcost. M Todd aEMR HCL/UCBL 3738 CTO, FacultedeMedecine Lyon-Sud, Universitede Keywords: intravenous paracetamol, indication, dose, inappropriate Lyon, Oullins, France; bLaboratoire de pharmacologie-toxicologie, Service de Pharmacie, Hôpital Cochin, Paris, France; cLaboratoire de pharmacologie-toxicologie, Service de Pharmacie, Hôpital Cochin, Paris, France; dEMR HCL/UCBL 3738 CTO, Facultede 22-10 Medecine Lyon-Sud, Universite de Lyon, Lyon, France Cardiovascular and neurological adverse drug reactions with oral or Objectives: Hand-Foot Syndrome (HFS) is a dose-limiting toxicity of sorafenib. nasal decongestants in France Its physiopathological mechanism has not been fully understood yet, but accu- E Bondon-Guitton, JL Montastruc, The French Network Of Regional mulation of a toxic compound in skin cells had been suggested. The objective of Pharmacovigilance Centers Service de Pharmacologie Clinique, Toulouse, France our work was to quantify the risk dynamics for the sorafenib-induced HFS and to Objective: Nasal or oral decongestants are used in the common cold. Nasal explore by simulations the dose-toxicity relationships according to different dosing decongestants are prescription drugs. Oral forms are over-the-counter drugs and regimen. available without any prescription. The French Drug Agency (ANSM) warned in Patients & Methods: Eighty-nine patients treated with sorafenib were consid- December 2011 about ‘serious’ cardiovascular and neurological adverse drug ered: treatment duration and regimen, and number and frequency of HFS obser- reactions (myocardial infarctions, hypertension, cerebral stroke…) with these vations were highly variable. drugs. Our aim was to describe the cardiovascular and neurological adverse drug A non-linear mixed effect model was built to link sorafenib administrations to the reactions (ADRs) reported in France with nasal or oral decongestant since the risk of each HFS score. The drug, whose PK had been described by a saturable previous national study performed in 2007. absorption one-compartment model, was considered as impacting the kinetics of Methods: We selected all cases registered in the French Pharmacovigilance Data- a latent variable (LV), interpretable as a non-identified biomarker. The probability base between the 25/11/2007 and the 31/08/2012 in which an oral (pseudo- of each HFS score is computed from a probit function of the LV level and corre- ephedrine or phenylephrine) or nasal (oxymetazoline, naphtazoline, sponding threshold parameters. Parameters were estimated in NONMEM7.1.2. phenylephrine, ephedrine and tuaminoheptane) decongestant was suspected in Model evaluation was driven by goodness-of-fit and simulation-based diagnostics. the occurrence of cardiac, vascular or neurological ADRs. Model simulations were performed considering the evolution of HFS risk over Results: Between the 25/11/2007 and the 31/12/2011, cardiovascular (39 42 weeks in 100 replicates of populations of 100 patients, receiving a sorafenib cases) or neurological (72 cases) ADRs, reported with an oral or nasal decon- daily dose ranging from 200 to 2000 mg, split over 1, 2, 3 or 4 occasions. gestant, were mainly « serious ». One patient died, a 83 years old male who Results & Discussion: A physiologically coherent model, relating sorafenib was concomitantly treated with fluindione and suffered from an abdominal administrations and, per se its exposure, to HFS dynamics has been built. hematoma. Misuse was found (simultaneous use of oral and nasal deconges- From the model simulations, it appears that the more the daily dose is fractioned, tants, use in children under 15, off-label dose and duration of treatment) in 10 the more the patients are at risk for HFS. For example, administering 1200 mg/ cases of cardiovascular ADRs and in 29 cases of neurological ADRs. Cardiovas- day for 42 weeks, the simulated median percentage of patients experiencing at cular and neurological ADRs were more frequently notified with oral deconges- least one severe HFS event were respectively 44%, 63%, 73% and 78% when the tants (1080 9 10À6 EIs per sold box) than nasal (0649 9 10À6 EI per sold daily dose is taken on 1, 2, 3, or 4 occasions. Interestingly, the number of daily box). Between the 01/01/2012 and the 31/08/2012, after the 2011 warning occasions was found more influential than the dose itself. from ANSM, cases still occurred: 9 cardiovascular cases and 18 neurological Understanding the dynamic relationship between drug administrations and an cases. induced adverse event is essential to control toxicities and adequately adjust Discussion: This survey still observed cardiovascular and neurological ADRs treatment modalities. Our original pharmacokinetic-pharmacodynamic model for with oral or nasal decongestant in France, mainly « serious » and more fre- sorafenib-induced HFS, including the kinetics of a latent variable, can be used as quently observed with the oral route. Such a rare but ‘serious’ risk appears to be an early predictor of severe toxicity risk in patients. unacceptable for common cold, a benign pathology. Keywords: sorafenib, hand-foot syndrome, modeling, simulation, dosing regimen Keywords: oral, nasal, decongestant, cardiovascular, neurological, adverse drug reactions 23-02 Increasing the reliability of the dose adjustment of cyclosporine by using 22-11 a combination of 3 Pharmacokinetic tools in Hematopoietic Stem Cell Adverse effects associated with the use of intravenous nicardipine. transplant patients JB Woillarda, V Lebretona, M Neelyb, P Turlurec, S Girautc, P Marqueta, Analysis of data from the French Pharmacovigilance database a a J Moragnya, V Grasa, S Chaplaina, E Drigneia, M Andrejaka aCentre de F Saint-Marcoux CHU LMOGES, Service de Pharmacologie, Toxicologie et pharmacovigilance de Picardie - CHU AMIENS, AMIENS and the French Pharmacovigilance; INSERM UMR-S580, Univ. LIMOGES, Limoges, France; bLaboratory of Apllied Pharmacokinetics, University of Southern California, Los Pharmacovigilance network c Objective: Intravenous (i.v) nicardipine is indicated in various hypertensive Angeles, CA, USA; CHU Limoges, Service d’Hematologie, Limoges, France emergencies which are defined as severe acute blood pressure elevations associ- Introduction: Cyclosporine A (CSA) is an immunosuppressive drug used in the ated with end-organ dysfunction and during surgical procedure to control intra- prophylaxis and treatment of acute and chronic graft-vs.-host disease after hae- and post-operative blood pressure when haemostasis may be difficult. This drug matopoietic stem cell (HCT) transplantation. The objective of this study was to is also used off-label in preterm labor, especially in France. create and compare three independent Bayesian estimators (BE) of CSA AUC from a limited sampling strategy, to assist in the management of CSA therapy.

Methods: The BE were created with nonlinear mixed-effect modeling in NON- nephrectomy 85.4%. Baseline sunitinib dose was 50 mg/day for 83.4% of MEMâ and iterative two stage (ITS) Bayesian in a home-made program as two patients. The median duration of first-line treatment was 10.7 months (median independent parametric population modeling approaches, and the nonparametric number of cycles: 6). Dose reduction occurred in 65.2% of patients. Reasons for adaptive grid in the Pmetrics package for R as a nonparametric approach. For discontinuation of first-line treatment were progressive disease (61.1%), death each approach, 72 full pharmacokinetic (PK) profiles collected from 40 HCT (31.2%), adverse events (6.8%), and other reasons (1.0%). OS was 70.8% at patients given CSA were used to build the PK model, while 15 PK profiles 1 year [95%CI (65.3; 75.5)] and 49.5% at 2 years [43.7; 55.0]. Median OS was (obtained from seven patients) were kept for validation. For each BE, fitted AUCs 23.6 months [19.9; 27.4] (not reached in patients with clear-cell component). from the full profiles were compared to AUCs fitted to only the CSA concentra- PFS was 38.2% at 1 year [32.7; 43.7] and 16.4% at 2 years [12.5; 20.9]. Med- tions just before (C0), one (C1) and four (C4) hours after a dose. ian PFS was 8.4 months [7.6; 9.9] [9.5 months (8.1; 11.0) in patients with Results: The PK profiles were fitted to a one-compartment model with first-order clear-cell component]. Overall best response was 31.1% (2.3% complete, 28.8% elimination combined with a gamma function for the absorption phase in ITS partial), 40.1% had stable disease and 18.5% progressive disease. and Pmetrics or an Erlang distribution in NONMEM. The derived BEs based on a Discussion: Despite certain differences regarding severity of cancer, these results C0-C1 h-C4 h sampling schedule accurately estimated AUC: mean Æ SD relative demonstrated that effectiveness of sunitinib in mRCC is close to efficacy reported bias of 2.05% Æ 13.31%, RMSE = 13.02% for NONMEM; 4.61% Æ 10.56, in the pivotal clinical trial [median OS: 26.4 months (95%CI 23.0; 32.9), median RMSE = 11.20% for ITS; and 0.30% Æ 10.12, RMSE = 10.47% for Pmetrics. PFS: 11.0 months (11.0; 13.0)], specially in patients with clear-cell component. Only 2, 1 and 1 out of the 15 estimated AUCs had an absolute bias >20% for Keywords: Pharmacoepidemiology; Sunitinib; Metastatic renal cell carcinoma NONMEM, ITS and Pmetrics, respectively. Additionally, the combination of the three results led to a pertinent dose proposal in the 15 patients. Keywords: Hematopoietic stem cell transplantation, therapeutic drug monitor- 24-02 ing, modeling, pharmacokinetic, bayesian estimation Relationship between drug-drug interactions, Multidrug Resistance associated Protein 2 (MRP2) function and methotrexate clearance in adults with lymphoid malignancies 23-03 IBENZ- De Bretagnea, A Le Gougeb, N Zahrc, JS Hulotc, V Leblondd, K Hoang- Adalimumab pharmacokinetics and concentration-effect relationship in Xuane, E Gyanf, S Lissandref, S Choquetd, C Le Guelleca aCHU de Tours; EA4245 rheumatoid arthritis Universite Francßois Rabelais de Tours, Tours, France; bCentre d’Investigation Clinique, D Ternanta, P Fuzibetb, E Ducouraub, O Vittecoqc, T Lequerrec, P Goupillea, INSERM202, Tours; c Service de Pharmacologie, Pitie-Salpêtriere, Paris, France; D Mullemana, G Paintauda aCNRS UMR 7292 GICC, Tours, France; bService de dService d’Hematologie, Pitie-Salpêtriere, Paris, France; eService de Neurologie, Pitie- rhumatologie, Tours, France; cService de rhumatologie, Rouen, France Salpêtriere, Paris; f Service d’Hematologie et Therapire Cellulaire, Tours, France Introduction: Adalimumab, an anti-TNF-a monoclonal antibody, is effective in Introduction: Methotrexate (MTX) is mainly eliminated via renal excretion active rheumatoid arthritis (RA). Subcutaneous injections of adalimumab lead to including tubular secretion through membrane transporters (OAT1, OAT3, highly variable concentrations between patients and this pharmacokinetic (PK) MRP2, MRP4, BCRP). Severe toxicities often occur in patients who do not elimi- variability partly explains the variability of clinical effect. However, adalimumab nate the drug well. Transporters’ function is under genetic control and can be PK after subcutaneous injection has never been reported. The goal of this study inhibited by co-administered drugs. We showed previously in healthy subjects is to build a simplified PK model for therapeutic drug monitoring (TDM) of ada- that the urinary coproporphyrin I/(I+III) ratio [UCP I/(I+III) ratio] was dependent limumab in CD patients and to describe the concentration-effect relationship of on ABCC2 polymorphisms and may reflect MRP2 activity. The aims of this pro- adalimumab in these patients using PK-PD modeling. spective population pharmacokinetic study were to analyze whether UCP I/(I+III) Methods: Adalimumab PK data were taken from a multicentric observational ratio, ABCC2 polymorphisms and drug-drug interactions may influence MTX study. Adalimumab 40 mg was administered subcutaneously every other week. clearance (CLMTX) in adult patients suffering from lymphoid malignancies. Trough adalimumab concentrations, CRP levels and RA disease activity score Patients and Methods: Patients were followed up the first MTX infusion during (DAS28) were measured at inclusion visit, then at weeks 6, 12, 24 and 52. Ada- which therapeutic drug monitoring of MTX was performed. A whole blood sam- limumab PK was decribed using a one compartment model with first-order ple was drawn for ABCC2 genetic testing. Three urinary samples were collected absorption and elimination rates. The relationship between adalimumab concen- for UCP I(I+III) ratio determination: one before MTX infusion (P1) as basal value, trations and CRP levels was described using using an indirect response model one at the end of MTX infusion (P2) and one the day of hospital discharge (P3). with inhibition of CRP input, whereas the relationship between adalimumab con- All associated drugs were exhaustively recorded and a drug interaction score was centrations and DAS28 was described using a direct inhibitory model. A popula- affected: a drug known to interfere with MTX elimination was set at2, a drug tion approach was used and models were run simultaneously. Sex, age and body known to interact with MTX transporters, at 1 and a drug with no interaction, weight were tested as covariates on each pharmacokinetic and PK-PD parameter. at zero. Toxicities were recorded and graduated according to standard criteria. Results: Thirty patients were included, and 131 adalimumab trough concentra- Results: Eighty-one patients were included. The basal UCP I/(I+III) ratios pre- tions, CRP levels and DAS28 measurements were available. The following PK sented a wide interindividual variability and was influenced neither by ABCC2 and PK-PD parameters were estimated (interidividual coefficient of variation): polymorphisms nor co-administered drugs. This ratio increased at P2 and apparent volume of distribution (Vd/F) = 12.4 L (75%), apparent clearance (CL/ decreased at P3 (P < 0.001). CLMTX was not correlated with basal UCP I/(I+III) F) = 0.31 L/day (17%), first-order absorption constant (ka) = 0.41 per day, zero- ratios but was lower in patients receiving at least one score 2 drug order CRP input rate (kin) = 129 mg/L/day (47%), first-order CRP output rate (CL=6.2 Æ 1.4 L/h vs. CL=7.3 Æ 1.8 L/h, P = 0.01). The creatinine clearance (kout) = 8.3 per day and adalimumab concentration leading to 50% decrease of and the variation of the UCP I/(I+III) ratio between P1 and P3 were also associ- kin (C50) = 5.7 mg/L (102%), initial DAS28 (DAS0) = 5.5 mg/L (11%) and ada- ated with CLMTX. Delayed elimination was associated with a higher risk of toxic- limumab concentration leading to 50% decrease of DAS0 (IC50) = 11.8 mg/L ity [OR = 4.9, (1.3–18.4), P = 0.02]. (66%). Apparent clearance was higher for increasing body weight and for men. Conclusion: Our study showed that drug interactions involving transporters Conclusion: This is the first study describing the pharmacokinetics and concen- may influence MTX elimination. Even if UCP I/(I+III) ratio was not correlated tration-effect relationship of adalimumab in rheumatoid arthritis. This model with CLMTX, its variation during MTX infusion confirms that it can be modulated allows therapeutic drug monitoring of adalimumab in RA patients. by substrates of MRP2. Keywords: adalimumab, rheumatoid arthritis, pharmacokinetics, concentration- Keywords: Methotrexate, drug transporters, MRP2/ABCC2, benzimidazole, drug- effect relationship, therapeutic drug monitoring drug interactions, population pharmacokinetics

24-03 SESSION 24: GT ONCO-PHARMACO Severe fluoropyrimidines toxicities: screen effectively for DPD deficiencies M Boisdron-Cellea, O Capitaina, JP Metgesb, T Lecomtec, T Matysiak-Budnikd, 24-01 A Morela aInstitut de Cancerologie de l’Ouest, Angers, France; bCHU Brest, Brest, Real-life patterns of use and effectiveness of sunitinib in patients with France; cCHRU Tours, Tours, France; dCHU Nantes, Nantes, France metastatic renal cell carcinoma: the SANTORIN study Background: Severe, even fatal toxicities which occur during the first course of A Grelaud-Boussinota, A Ravaudb, JO Bayc, C Bairras-Martina, A Balestraa, fluoropyrimidines chemotherapy treatments pose a serious public health problem. C Chevreaud, MA Bernarda, E Bignona, S Culinee, S Lamarquea, R Lassallea, In adjuvant therapies, using fluoropyrimidines engenders a toxic (even fatal) risk M Rouyera, N Mooref, P Noizeb, A Fourrier-Reglatf aUniversite Bordeaux Segalen, to a patient who is possibly in remission, if not cancer-free. INSERM CIC-P0005, Bordeaux, France; bCHU de Bordeaux, INSERM CIC-P0005, Method: From April 2001 to December 2011, 11 351 patients were screened Bordeaux, France; cCHU de Clermont-Ferrand, Site Estaing et Centre Jean Perrin, for the risk of severe toxic reaction to fluoropyrimidines. Chemotherapy protocols Clermont-Ferrand, France; dInstitut Claudius Regaud, Toulouse, France; eCHU Saint were 5-Fluorouracil based, as well as orally- delivered fluoropyrimidines. 11 104 Louis, Paris, France; fUniversite Bordeaux Segalen, CHU de Bordeaux, INSERM patients were screened before treatment, as has been the regular protocol in our U657, INSERM CIC-P0005, Bordeaux, France institution. 247 blood samples were received from other institutions from patients Objective: Until 2006, treatment options for metastatic renal cell carcinoma who, after having received their first treatment, showed signs of severe (grade III- (mRCC) were limited to cytokine therapy and surgery. Sunitinib, an oral antian- IV) toxicity, or had died. The evaluation of risk associates genotyping for muta- giogenic agent, received European marketing authorisation in January 2007 for tions on the DPYD gene and phenotyping (dihydrouracil and plasmatic uracil) by first-line treatment in advanced and/or mRCC. The SANTORIN study was HPLC. The calculations were made via the ODPMTox, and every patient whose designed to describe patterns of use and estimate survival outcomes in mRCC screening before treatment showed risk received an individually-adapted dose via patients treated with sunitinib in a real-life setting. ODPMProtocol. Methods: SANTORIN was an observational cohort conducted in 36 French cen- Results: In the patient population screened after toxicity occurred (247 tres including patients initiating first-line treatment with sunitinib from January patients): 2008 to April 2010. Patients were followed 2 years after treatment onset. Eleven percentage died from toxicity. 59% had one or more genetic mutations Treatment characteristics, response (evaluated by investigator based on RECIST and 5% presented with no mutation, but a phenotype characteristic of DPD defi- criteria) and survival outcomes were collected from medical files. Overall and pro- cit according to ODPMToxTM criteria. Thus, the technique has over 99.9% reliabil- gression-free survival (OS and PFS) results were estimated using Kaplan-Meier ity. 89% showed grade III – IV toxicity after first treatment. Among these method. patients, 30% presented with genetic mutation and 67% showed altered pheno- Results: Characteristics of the 302 included patients were: mean age 64.3 years; type (ODPMToxTM). In population screened before treatment (11, 104 patients). male 73.2%; clear-cell histological component 83.1%; ECOG 2 9.9%, prior No patients died and none presented with grade III-IV toxicity. Among this popu-

lation, 2.4% patients carried one or more risk-related mutations and 8.6% SESSION 25 : PHARMACOLOGIE EXPERIMENTALE, showed a phenotype characteristic of DPD deficiency. All of these patients received chemotherapy treatments using the ODPMToxTM dose-adjustment and all CELLULAIRE ET MOLECULAIRE subsequent rounds were dose-adjusted using ODPM ProtocolTM. Conclusion: The dual approach to DPD deficiency screening using genotype and phenotype before treatment avoided all severe toxicities. This system has been 25-01 used in our institution for the past 10 years on a routine basis. All of the Inhibitors of the protease-activated receptor-1 reduce the atrial remodelling in a rat model of chronic heart failure patients who carried a known risk for toxicity were either treated by another a a b c a a protocol without fluoropyrimidines, or the 5-Fluorouracile treatments were dose- C Jumeau , P Chieng-Yane , B Le Grand , S Hatem , M Dufilho UMRS 956 Inserm-UPMC, cedex 13, Paris, France; bCentre de Recherche Pierre Fabre, Castres, adjusted. c Keywords: DPD deficiency, Toxicities, fluoropyrimidines France; UMRS 956 Inserm-UPMC, Paris, France Atrial fibrillation (AF) complicates the atrial remodelling characterized by myocardial fibrosis and dilation. AF is often associated with an activation of the endo- 24-04 cardial endothelium that leads to the accumulation of thrombin in the atrial cavity and subsequent formation of micro-thrombi. The aim of the present study Study of HSP110DE9 in 86 patients with metastatic colorectal cancer a b a a b a was to examine the participation of thrombin and its protease-activated receptor- C Bris , A Morel , C Renaudin-Fonsegrive , B Linot , M Boisdon-Celle Institut de 1 (PAR-1) in the atrial remodelling associated with chronic heart failure (HF) in cancerologie de l’ouest, site Paul Papin, Angers, France; bInstitut de cancerologie de rat. l’ouest, site Paul Papin, et Unite INSERM U892, Angers, France Methods: Rats underwent either thoracotomy only (SHAM) or thoracotomy with Purpose: Colorectal cancer (CRC) with mismatch repair deficiency (dMMR), transient ligation of the left coronary artery (HF). To evaluate the role of PAR-1 characterized by high-frequency microsatellite instability (hMSI), seems to present in the development of the cardiopathy, a part of rats were treated with daily oral a better prognostic, although they are less receptive to 5-Fluorouracil (5-FU) administration (5–40 mg/kg) of PAR-1 antagonists (F16618 Pierre Fabre, based chemotherapy. Recently, a deletion in an intronic microsatellite of HSP110 D SCH203099 Schering). Rats were then sacrificed 2 months after surgery. The gene (T17) was found to lead to a truncated protein, HSP110 E9, in hMSI cardiopathy was assessed by echography and histology. Expression of remodelling tumors. HSP110DE9 does not show chaperon activity and inhibits HSP110’s D and endothelial markers and the signalling mediators of PAR-1 were explored by functions. HSP110 E9/HSP110 rate upper to 75%, detected in 30% of the hMSI quantitative RT-PCR, western blot and immunofluorescence. tumors, seems to sensitize cells to 5-FU. This study aimed at investigating the D Results: In HF group, there was a clear left ventricle (LV) dysfunction as indi- expression and impact of HSP110 E9 in metastatic CRC (mCRC). cated by the marked LV dilation together with the reduced shortening fraction Methods: We selected 86 mCRC patients and treated by 5FU, irinotecan and ce- 56%. Left atria diameter and atria weight/heart weight were increased by 50% tuximab. MSI status was assessed for each tumor by molecular genetics (MSI â and 75%. Histological study revealed hypertrophic trabeculae with interstitial Analysis System, Promega ) and/or immunochemistry. The length of T17 was fibrosis. The endothelial activation was indicated by the increased expression of evaluated on constitutional and tumor DNA by PCR and fragment analysis â D the pro-thrombotic factor vWf at both mRNA and protein levels. Atrial remodel- (Genetic analyser 3130, Applied ). The HSP110 E9/HSP110 ratio was deter- ling was indicated by increased mRNA of b myosin heavy chain (bMHC) and mined by RT-PCR and fragment analysis. Response to chemotherapy, overall sur- brain natriuretic peptide (BNP). Dedifferentiation and apoptosis of atrial myocar- vival (OS) and disease free survival (DFS) were studied retrospectively. dium was shown by the up-regulation of a1 actin and caspase 3. This was asso- Results: We did not observe any polymorphic variations of T17 in constitutional ciated with amplified expression of PAR-1 and its activator MMP-1. Both F16618 DNA: 67% and 23% of patients had 16 repetitions, 17 repetitions of thymidine, and SCH203099 treatments reduced the atrial dilation but they had no signifi- respectively. Compared to healthy tissues, all 6 MSI tumors presented a T17 dele- – D cant effect on the LV dysfunction and dilation. Administrated at 10 mg/kg, tion (1 7 bp), whereas, in MSS tumors, T17 was unchanged. The HSP110 E9/ F16618 prevented the increase in the expression of bMHC, BNP, skeletal actin, HSP110 ratio was determined in 4 MSI tumors and one was upper 75%. D caspase 3 and PAR-1. However, treatment with F16618 had no significant effect HSP110 E9 was not detected in MSS tumors. No significant difference was on the expression of vWf and MMP-1. observed between patients with MSI ou MSS tumors in terms of response to che- Conclusions: Altogether, these data demonstrate that the PAR-1 antagonist motherapy, OS and DFS. F16618 is highly effective to prevent atrial remodelling associated with HF devel- Conclusion: This study confirmed the exclusive presence of HSP110DE9 in MSI D opment. tumor. HSP110 E9/HSP110 ratio is variable (21% to 116%) and seemed not to Keywords: Heart failure - Atrial remodeling - Protease activated receptor-1 – be correlated to T17 length. However, because of the small number of MSI Antagonist tumors, the influece of HSP110DE9 on response to chemotherapy, DFS and OS was not evaluated Further investigations will be made to explain the differences in HSP110DE9 expression and to assess their impact on the response to 5-FU based chemotherapy on a larger sample of MSI tumors. 25-02 Mechanisms involved in the increase mBDNF after rt-PA infusion? Keywords: Colorectal cancer, Microsatellite instability, 5-fluorouracil, a a a a a HSP110DE9 M Rodier , C Mossiat , C Marie , P Garnier INSERM U1093 Action, Cognition et Plasticite Sensorimotrice, Dijon, France Recombinant tissue plasminogen activator (rt-PA) is the only curative approach 24-05 in the treatment of ischemic stroke. rt-PA cleaves the plasminogen into plasmin that in turn dissolves the clot of the occluded vessels. In addition to its fibrinolytic Pharmacokinetic interactions between antiretroviral and antineoplastic role, it has been recently shown that plasmin can also cleave the pro-BDNF into agents: elaboration of a synthetic document mature BDNF (mBDNF), a neurotrophin that plays a central role in many facets A Disson-Dautrichea, S Combreta, A Grandvuillemina, S Pierrea, C Flecka, a a of neuroplastic processes. In addition, recent studies have demonstrated that rt- C Sgro Centre Regional de Pharmacovigilance de Dijon, Dijon, France PA can activate NMDA receptors that lead to neuronal excitation that can be fol- Background: Malignancy has become one of the major causes of morbidity and lowed by BDNF release. mortality in HIV-infected patients. However antiretroviral and antineoplastic We recently showed that administration of rt-PA induces an increase of mBDNF agents have many pharmacological interactions. The requests of the practitioners expression in the hippocampus 2 h and 24 h after the beginning of perfusion. to Regional Centres of Pharmacovigilance seem more and more frequent and Based on these results, our study was designed to delineate the mechanisms appear difficult because of the complex pharmacological profile of the molecules involved in this process. For this purpose, we use a plasmin inhibitor: the tra- and multiple documentary sources not always easy to exploit. nexamic acid, and an antagonist of NMDA receptor: the MK801. Our experi- Objectives: To elaborate a practical and useful tool in order to answer clinicians ments were carried out on male Wistar rats (290–310 g). Animals were treated about pharmacokinetic interactions between antiretroviral and antineoplastic with tranexamic acid (300 mg/kg, i.v.) Æ rt-PA (10 mg/kg, i.v.) or with drugs. MK801 (3 mg/kg, i.p.) Æ rt-PA (10 mg/kg, i.v.) or with vehicle. Tranexamic Methods: A bibliographic synthesis was realised. For each antiretroviral drug, acid, rt-PA and vehicle were administrated over a period of 1 h, while MK801 we looked after its metabolic, transport and elimination pathways, and its poten- were injected by bolus. The animals were euthanized 2 h or 24 h after the tial clinical drug interactions with antineoplastic drugs. The data were presented beginning of the administration. Using a new antibody recognizing the mature as a table. form of BDNF, mBDNF expression was assessed by Western blotting in the Results: We found many potential drug interactions via the cytochrome P450 hippocampus. enzyme system and/or the multidrug transporter P-glycoprotein, the UDP-glucu- First of all, as compared to control animals, control tranexamic acid or MK801 ronosyltransferase 1A1 (UGT1A1) or via a common renal elimination pathway. groups exhibited no variation in term of mBDNF expression and thus whatever the Thus, among these interactions, protease inhibitors (IP) inhibit the cytochrome time points considered. Furthermore, when injected during rt-PA administration, P450 isoenzyme CYP3A4, metabolic pathway of numerous substrate antineoplas- the plasmin inhibitor does not reverse the increase of mBDNF induced by rt-PA at tic drugs including tyrosine kinase inhibitors, irinotecan, vinca-alcaloids with a 2 h and 24 h. However, according to our preliminary results, MK801 seems to risk of overdose and toxicity. This effect is well described with ritonavir, used as a counteract the increase of mBDNF expression induced by rt-PA administration. pharmacokinetic enhancer of other IP. Atazanavir inhibits CYP2C8 and UGT1A1 In conclusion, our results suggest that the rise of mBDNF expression in the hip- and is able to increase paclitaxel and irinotecan levels. This interaction seems pocampus after rt-PA perfusion is plasmin-independant, but may be NMDA- particularly at risk for irinotecan. Non nucleoside reverse transcriptase inhibitors dependant. have also drug interactions via induction or inhibition of the hepatic oxidative Keywords: BDNF, tissue plasminogen activator, NMDA, plasmin, tranexamic system. Nucleoside reverse transcriptase inhibitors, have a renal elimination as acid, MK801, Hippocampus methotrexate, pemetrexed and topotecan. Thus, attention must be paid to these drug combinations. Unlike, maraviroc, a CC chemokine receptor 5 antagonist, raltegravir, an inhibitor of HIV-1 integrase, and enfuvirtide, a fusion inhibitor, seam not to fear drug interactions. 25-03 Conclusion: we present a synthetic document in order to identify the main and Implication of SIRT1 in beneficial effects of PARP inhibition by 3-aminobenzamide during in vivo cerebral oxidative stress potentially serious pharmacokinetic dug interactions in HIV-infected patients a a a a a a treated for cancer and facilitate the requests of the practitioners. This process C Gueguen , B Palmier , M Plotkine , C Marchand-Leroux , V Besson EA4475 emphasized the importance of a multidisciplinary care of these patients involving - Pharmacologie de la Circulation Cerebrale - Universite Paris Descartes, Paris, France infectiologist, oncologist, and pharmacologist. Introduction: Poly(ADP-ribose)polymerase (PARP) and sirtuin1 (SIRT1) are Keywords: antineoplastic drug, antiretroviral drug, drug interaction, pharmaco- both NAD-consuming enzymes. In vitro oxidative stress (OS) activates PARP, kinetic

decreases NAD level, SIRT1 activity and finally leads to cell death. PARP hyper- by a high level of anxiety in HFD mice as shown by a decrease in the time spent activation contributes to cell death and its inhibition protects against in vivo in the anxiogenic areas (402 Æ 41 vs. 701 Æ 50 s. in the center of the arena; cerebral OS. NAD level and SIRT1 activity are restored in vitro by PARP inhibi- P < 0.001 and 29 Æ 3 vs. 42 Æ 6 s. in the open arms; P < 0.05) in the open- tion. Furthermore, SIRT1 induction protects neurons from cell loss induced by field and the elevated plus maze tests; respectively. Another symptom of depres- OS in vitro. In this context, the aim of our study was to evaluate in vivo the sion such as helplessness reflected by a decreased immobility time in the tail involvement of SIRT1 on histological and functional consequences induced by suspension test was also unveiled in HFD compared to STD mice (120 Æ 12 vs. cerebral OS and its role in the beneficial effects of PARP inhibition during 72 Æ 10 s; P < 0.05). Finally, in the splash test measuring the time of grooming cerebral OS. after the application of sweetened solution on the coat, this parameter was Materiel & Methods: In vivo cerebral OS was induced in male Sprague-Dawley decreased in HFD compared to STD mice (95 Æ 7 vs. 180 Æ 10 s; P < 0.001). rats by intrastriatal infusion of malonate. The PARP inhibitor 3-aminobenzamide This result indicated a carelessness frequently observed in depressed patients. (3AB), the SIRT1 activator SRT1720, the SIRT1 inhibitor EX527, the association These data support the idea that T2D may precipitate depression. Experiments 3AB+SRT1720, the association 3AB+EX527 and their vehicle, were injected in- are in progress to test the antidepressant-like response of chronic administration tracerebroventricularly 30 min before malonate infusion. Neurological assess- of SSRIs in HFD and STD mice. This aspect is important for the ‘well-being’ of ment, measured by a score on 15 points, and brain lesion were evaluated at 6 h diabetic-depressed patients. Indeed, an early and adapted psychiatric care may after malonate. improve the efficacy of their anti-diabetic treatment by a better style of life and Results: Malonate decreased the score (5.7 Æ 0.5 vs. 14.4 Æ 0.2 for non-oper- compliance. ated rats; P < 0.001), showing a neurological deficit, associated with a striatal Keywords: diabetes, depression, anxiety, antidepressant lesion (46 Æ 1mm3). Both were reduced by 3AB (score: 9.4 Æ 0.6, lesion: 30 Æ 5mm3; P < 0.01) and SRT1720 (score: 11 Æ 1, P < 0.01; lesion: 32 Æ 4mm3, P < 0.05), showing beneficial effects of PARP inhibition and SIRT1 25-06 activation on OS consequences. Inhibition of SIRT1, by EX527 given alone, modi- Fludrocortisone and hydrocortisone, alone or in combination, on in vivo fied neither the score nor the lesion, demonstrating that SIRT1 inhibition did not hemodynamics and in vitro vascular reactivity in normal and exacerbate the neurological and histological consequences of OS. However, its endotoxemic rats association with 3AB suppressed the neurological improvement (5.6 Æ 0.5; a a a a a 3 B Laviolle , N Nesseler , C Massart , E Bellissant CHU de Rennes, Universitede P < 0.001) and the reduction of striatal lesion (47 Æ 3mm; P < 0.01) induced Rennes 1, CIC Inserm 0203, Rennes, France by 3AB. The association of 3AB with SRT1720, the SIRT1 activator, did not Objectives: Single administration of a physiologic dose of hydrocortisone modify the reduction of neurological deficit (8.6 Æ 1) and striatal lesion 3 enhances the pressor response to catecholamines in healthy volunteers and septic (37 Æ 4mm) induced with 3AB alone. shock patients [1]. Similar data do not exist for fludrocortisone [2, 3]. We Discussion: For the first time, we showed that SRT1720, a SIRT1 activator, pro- assessed the effects of single administrations of fludrocortisone and hydrocortisone tected from the consequences of OS. The fact that a SIRT1 inhibitor suppresses on in vivo hemodynamics and in vitro vascular reactivity in normal and endo- the protective effect of 3AB indicates that SIRT1 activation plays a major role in toxemic rats. the protective effect of PARP inhibition. Methods: Intravenous fludrocortisone (5 and 20 lg/kg) and hydrocortisone (4 Keywords: oxidative stress, poly(ADP-ribose)polymerase, sirtuin 1, cell death and 20 mg/kg) were administered in 16 groups (8 without and 8 with lipopolysaccharide-induced endotoxemic shock) of 10 Wistar rats who respectively received placebo, fludrocortisone dose 1 or 2, hydrocortisone dose 1 or 2, or flu- 25-04 drocortisone + hydrocortisone dose 1 or 2, according to four 2 9 2-factorial Role of 15-lipoxygenase in the polarization of human lung macrophages designs. Systolic blood pressure was recorded at baseline, after lipopolysaccharide C Abriala, S Grassin Delylea, M Brolloa, E Nalinea, P Devilliera aLaboratoire UPRES or saline injection, and 2 times after treatment administration. At 3 h, the EA220, Suresnes, France response of mesenteric artery rings to cumulative doses of phenylephrine was Introduction: Derivates from the 15-lipoxygenase (15-LO) pathway have been measured in isolated organ baths. involved in the pathogenesis of airway allergic inflammation, and 15-LO products Results: Fludrocortisone and hydrocortisone similarly increased systolic blood increase the production of pro-inflammatory cytokines by human lung epithelial pressure 10 min after their administration (P < 0.001 for both) but more in en- cells or murine macrophages. Moreover, 15-LO expression is up-regulated by IL-4 dotoxemic than in normal animals. Fludrocortisone and hydrocortisone similarly and IL-13 in human monocytes-derived macrophages. Because the functions of affected contractile response to phenylephrine (P = 0.039 and P = 0.009, respec- 15-LO in the M1 (pro-inflammatory) or M2 (immunoregulatory) macrophage tively). At dose 1, fludrocortisone had no effect and hydrocortisone decreased polarization have not been yet addressed, the aims of our work were to investi- contraction, whereas, at dose 2, both fludrocortisone and hydrocortisone gate the involvement of 15-LO in the polarization of human lung macrophages increased contraction, especially in endotoxemic rats. (LM). Conclusion: Single intravenous administrations of fludrocortisone and hydrocor- Methods: LM were isolated from patients undergoing surgery for carcinoma and tisone increase blood pressure and contractile response of mesenteric arteries to challenged for 24 h with 10 ng/mL LPS (to obtain M1 LM), 50 ng/mL IL-13 or phenylephrine. The magnitude of these effects depends on dose and pathophysio- 10 ng/mL IL-4 (to obtain M2 LM), treated or not with the dual 12-/15- or the logical conditions and is higher in endotoxemic than in normal rats. specific 15-LO inhibitors (nordihydroguaiaretic acid (NDGA) and PD146171, References: respectively). 15-LO transcript expression was determined with RT-qPCR. M1- 1. Bellissant E., Annane D. Effect of hydrocortisone on phenylephrine–mean arte- type (TNF-a, CXCL8) and M2-type (CCL18, CCL22) cytokines were quantified rial pressure dose-response relationship in septic shock. Clin Pharmacol Ther with ELISA, and the 15-LO metabolite 15(S)-HETE was assessed with EIA. (2000) 68 293–303 Results: Unstimulated LM were found to express 15-LO transcripts, which were 2. Laviolle B., Le Maguet P., Verdier M.C., et al. Biological and hemodynamic up-regulated (about 300-fold) after IL-13 or IL-4 exposure but unaffected after effects of low doses of fludrocortisone and hydrocortisone, alone or in combina- LPS. Accordingly, the basal production of the 15-LO metabolite 15(S)-HETE was tion, in healthy volunteers with hypoaldosteronism. Clin Pharmacol Ther (2010) increased by both IL-13 and IL-4 (about 135-fold), and NDGA prevented this 88 183–190 increase. NDGA (10 lM) and PD146171 (10 lM) also inhibited the IL-4/IL-13- 3. Laviolle B., Donal E., Le Maguet P., et al. Low doses of fludrocortisone and induced release of CCL18 and CCL22, as well as the release of TNF-a and CXCL8 hydrocortisone, alone or in combination, on vascular responsiveness to phenyl- in LPS-challenged cells. Inhibitory effects were 30–46% for M1-cytokines and 20 ephrine in healthy volunteers. Br J Clin Pharmacol (2012); [Epub ahead of print –67% for M2-cytokines. On the other hand, the cyclooxygenase-2 inhibitor indo- jun 15] methacin (1 lM) had no impact on the production of either M1 or M2 cytokines Keywords: Fludrocortisone, hemodynamics, hydrocortisone, inflammation, vas- in LM pre-treated with NDGA or PD146171. cular reactivity Conclusion: Altogether, our results show that the 15-LO pathway regulates the production M1- and M2-cytokines in human LM. The effect is not related to a shift of arachidonic metabolism towards the PGE2-producing cyclooxygenase 25-07 pathway (anti-inflammatory). 15-LO may be a new target for the pharmacologi- Cardiac ischemia increases interstitial beta-NAD, NAADP and cADP- cal management of lung inflammation. ribose levels Keywords: 15-lipoxygenase, lung macrophages, cytokines, macrophages polari- Z Djeradaa, S Dukicb, M Vittierb, H Millarta aDepartment of Pharmacology, zation E.A.3801, SFR CAP-sante, Reims University Hospital, Reims, France; bLaboratory of pharmacology and pharmacokinetic. Medyc-CNRS-3481-Reims University, Reims, France 25-05 Objective: Even if purine and pyrimidine derivatives as single nucleotides have Impact of type 2 diabetes on depressive-like phenotype and been the most widely studied P2 agonists, it appears interesting to study the role antidepressant response in mice of b-nicotinamide adenine dinucleotide (b-NAD+) and its metabolites in important B Guiarda, G Quesseveura, A Gardiera,LPenicaudb, X Fioramontib aLaboratoire de regulatory functions. We particularly focused on the role of extracellular pyridine Neuropharmacologie EA3544, Chatenay-Malabry, France; bCentre des Sciences du nucleotides in triggering cardioprotective effects against ischemia and reperfusion Gout^ et de l’Alimentation - Universite de Bourgogne, Dijon, France (I/R). To provide a logic to study their potential role during I/R, we report for the Evidence indicates that the prevalence of depression in diabetic subjects is higher first time, kinetic data related to pyridine nucleotides release such as b-NAD+ than that in the general population. Despite these observations, the mechanisms and their active metabolites, nicotinic acid adenine dinucleotide phosphate (NA- underpinning the link between metabolic and psychiatric disorders are poorly ADP) and cyclic adenosine diphosphate-ribose (cADP-ribose), during an ischemia- understood. Moreover, it has been proposed that an impairment of glucose reperfusion cycle. homeostasis may result in blunted antidepressant response to selective serotonin Methods: To study the release of endogenous nucleotides (NAADP, NAAD, reuptake inhibitors (SSRI). To confirm and better understand the impact of type cADP-ribose, NAD, NADP, UTP) and Adenosine into the interstitial compartment, 2 diabetes (T2D) on the physiopathology and treatment of depression, this study a perfused rat heart model (n = 5) was instrumented with cardiac microdialysis1. evaluated the effects of a long-term (8 weeks) high fat diet (HFD) on depressive- Probes were constantly perfused at 1 lL/min, and eluents were collected on ice like phenotype and SSRI response in mice. over 10-min periods. After a 40-min stabilization period, hearts were subjected to The results indicated that mice fed a HFD (protein 21% - fat 5%) exhibited a 40 min global ischemia and 40 min reperfusion. Dialysates were analysed using higher body weight (34.5 Æ 0.9 vs. 28 Æ 0.6 g; P < 0.001) and basal blood glu- ultra-performance liquid chromatography with tandem mass spectrometry detec- cose level (180 Æ 4 vs. 139 Æ 5 mg/dL; P < 0.001) associated with a moderate tion. Data were collected according to multiple reaction-monitoring modes. insulin resistance compared to control mice fed a standard diet (STD: protein Nucleotide concentrations were corrected by the mean in vitro recovery to yield 20% - fat 45%). The alteration of these metabolic parameters, were accompanied estimations of extracellular fluid concentrations.

Results: A time-dependent increase of all the abovementioned analytes was Conclusions: Anodal iontophoresis of A-350619, a sGC activator, increases observed during global ischemia. A fast recovery to basal values was observed cutaneous blood flow with good local tolerance while other tested drugs exhibited during reperfusion period. Comparatively to the basal value (before ischemia), the poor or no effect. Iontophoresis of sGC activators should be investigated as a new higher increase was observed with NAADP after 40 min of ischemia (0.8 vs. local therapy for digital ulcerations in patients with scleroderma. 0.03 lM, P < 0.001), followed by UTP (1.5 vs. 0.08 lM, P < 0.05), NAD (0.35 Keywords: iontophoresis; soluble guanylate cyclase; prostacyclin; non-prostanoid vs.0.03 lM), cADP-ribose (0.06 vs. 0.006 lM), and adenosine (2 vs. 0.23 lM, prostacyclin receptor; microcirculation P < 0.05). The increase of NAADP metabolite, NAAD, was non-significant (0.1 vs. 0.02 lM). Conclusion: We confirm that UTP and adenosine are released during ischemia. 25-10 To the best of our knowledge, this is the first report of a significant increase in Structure and activation mechanism of G-protein-coupled receptors: role extracellular concentrations of NAD and its metabolites such as NAADP and of the proline residues in helices 2 and 5 cADP-ribose during heart ischemia. These metabolites could be involved in V Chantreaua, L Gourdina, M Muniera, P Rodiena, M Chabberta aUMR CNRS endogenous cardioprotective mechanisms. 6214 - INSERM 1083, Angers, France Peart J and Headrick JP, Am J Physiol (2000). Objectives: Class A G-Protein-coupled receptors (GPCRs) constitute a large fam- Keywords: interstitial beta-NAD, NAADP and cADP-ribose, cardiac ischemia, ily of transmembrane receptors. Helical distortions play a major role in the over- microdialysis all fold and in the activation mechanism of these receptors. Most distortions are related to the presence of conserved proline residues. However, in helices TM2 and TM5, the presence of proline is not mandatory and the correlated mutation 25-08 of these proline residues is observed in several GPCR sub-families. In addition, the Chicoric acid is an anti-oxidant molecule stimulating AMP Kinase, PGC- position of the TM2 proline is variable (2.58–2.60). We are interested in the role 1alpha expression and mitochondrial activity in a model of skeletal of the TM2 and TM5 proline residues in the folding and activation mechanism of muscular cells GPCRs. A Schlernitzauera, C Oirya, F Casasb, B Chabib, G Crosa, R Magousa, G Cabellob, Methods: We selected two receptors, the vasopressin receptor 2 (V2R) and the C Wrutniak-Cabellob aCNRS UMR 5247, Institut des Biomolecules Max Mousseron thyrotropin receptor (TSHR), as prototypes of receptors with and without proline, IBMM, Universites Montpellier 1 et Montpellier 2, Montpellier, France; bINRA UMR respectively, in both TM2 and TM5. By site-directed mutagenesis, we engineered 866, Dynamique Musculaire et Metabolisme, Universites Montpellier 1 et Montpellier TSHR mutants with proline residues at different positions in TM2 and/or at posi- 2, Montpellier, France., Montpellier, France tion 5.50 in TM5. We also engineered V2R mutants without proline in TM2 and Introduction: It was suggested that food anti-oxidants may prevent insulin /or TM5 and with proline at different positions in TM2. We studied the influence resistance and development of oxidative stress associated with metabolic diseases. of these mutations on the receptor folding, membrane expression and activation In particular, caffeic acid and its derivatives, chorogenic and chicoric acids, were of the cAMP pathway. described as having anti-diabetic properties, although their mechanism is not Results: Most mutations in either TSHR or V2R impair the folding and the completely understood. In the present work, we used the L6 cell line model of translocation to the plasma membrane of the receptors. In particular, mutations skeletal muscle to explore the mechanism of chicoric acid (CA) by determining its in both helices are very disruptive. However, a few single mutations are tolerated effects on oxidative stress, michondrial activity, AMP-activated kinase (AMPK) and the corresponding mutants are able to activate the cAMP pathway with and insulin pathways. decreased EC50. Material and methods: Protein and mRNA expressions were determined by Discussion: Our experimental results enlighten the importance of the presence Western blotting and qPCR, respectively. Reactive oxigen species (ROS) were (or absence) of the TM2 and TM5 proline residues in the activation mechanism quantified using the 2’,7-dichlorofluorescein probe and cellular glucose uptake of specific GPCR sub-families. Coupled to molecular modeling, our study will help determined using 2-deoxy-D-glucose[3H]. understand the specific features of each receptor in relation with their proline Results: In differenciated L6 myotubes, CA was shown to be a ROS scavenger, pattern to improve drug design. both in basal and oxidative stress conditions. CA increased the activities of anti- Keywords: receptor, GPCR, proline, helix, activation mechanism oxidant defense system enzymes glutathion peroxydase and superoxyde dismutase (SOD). CA protected mitochondria against oxidative damages by increasing MnSOD expression. CA also increased the activity of complex II and the expres- 25-11 sion of PGC-1a, a transcriptional co-activator involved in the regulation of anti- New targets of endocrine disruptors: example of FSH receptor oxidant enzyme expression and mitochondrial biogenesis. CA also stimulated M Muniera, L Gourdina, V Chantreaua, M Chabberta, P Rodienb aInserm U1083 - AMPK/ACC and inhibited insulin-stimulated Akt/mTOR pathways without any Cnrs 6214, Angers, France; bCHU d’Angers, Angers significant change in glucose uptake. Research in the field of endocrine disruptors (ED) has tremendously expanded Conclusion: CA possesses anti-oxydant properties, both through its capacity to during the last years. In humans, ED was associated with sexual development neutralize ROS and increase anti-oxidant enzymatic defense systems. CA also abnormalities (genital malformation, precocious puberty). Regarding the mecha- stimulates mitochondrial biogenesis and protects mitochondria against oxidative nisms of action of ED, most of the published studies have focused on their inter- damages, along with increasing the mitochondrial capacity to oxidize fatty acids ferences with nuclear receptor signalization and/or with alterations in hormone by stimulation of AMPK and increase of complex II activity. Those results suggest synthesis. However, these target systems and axis partly depend on G protein that the potential of chicoric acid to prevent or treat metabolic syndrome-related coupled receptors (GPCRs) that are involved in numerous physiologic and physiopathologies deserves to be further studied. pathologic processes. In this context, we hypothesized that the GPCRs should be Keywords: Free Radicals, Mitochondria, Chicoric acid, myotubes, anti-oxidant, regarded as putative targets for ED. Initially we focused on the impact of ED on AMPK the FSH (Follicle-Stimulating Hormone) receptor (FSHR) activity. Molecules (pesti- cide, bisphenol A, phthalates, and heavy metals) are tested solo or in combination, to mimic multiple expositions in vivo, on stably transfected cell lines 25-09 overexpressing the human FSHR. The effect of ED on FSHR activity is assessed by Anodal iontophoresis of a soluble guanylate cyclase activator induces a cyclic AMP measurement by biosensor kit. The majority of pollutants tested sustained increase in skin blood flow in rats potentiate the maximal response of FSHR. Thus, the maximal response is a a b c d e À4 À8 S Kotzki , M Roustit , C Arnaud , J Boutonnat , S Blaise , D Godin-Ribuot , increased 1.5-fold with 10 M Mono Butyl Phthalate or 1.3-fold with 10 M JL Cracowskia aCentre d’Investigation Clinique - INSERM CIC03, Grenoble, France; bisphenol A. However, DiEthylHexyl Phthalate do not impact the FSH dose- b c À5 Inserm U1042 - Universite Joseph Fourier, Grenoble, France; TIMC UMRCNRS response in contrast to their metabolites such as 10 M Mono EthylHexyl Phtha- 5525, Grenoble, France; dDepartement de Medecine Vasculaire CHU Grenoble - late, that rise 2.3-fold the maximal response. Moreover, we observed a synergic INSERM U1042, Grenoble, France; eInserm U1042 - Universite Joseph Fourier, effect between ED and heavy metal. We now, try to determine the ED action Grenoble, France mechanisms and more particularly their impact on internalization, desensitization Introduction: The treatment of systemic sclerosis-related digital ulcers is still a or the recycling process of receptor. therapeutic challenge. While the only effective drugs are prostacyclin analogues, Keywords: Endocrine-disruptors; G protein coupled receptor; FSH receptor their current use is limited by frequent vasodilation-related adverse reactions. Iontophoresis is a non-invasive method permitting local drug-delivery, which may allow reaching high concentrations of drugs into the dermis while limiting their systemic effects. The objectives of this study were: 1/ to screen whether ion- SESSION 26: PHARMACO ET GROSSESSE tophoretically-administered non-prostanoid IP agonists and soluble guanylate cyclase (sGC) activators increase skin blood flow in rats; 2/ to test different drug concentrations and protocols, and 3/ the safety of candidates. 26-01 Methods: Three sGC activators (A-350619, SIN-1 and CFM 1571) and two non- Drug-induced adverse reactions via breastfeeding: a study in the French prostanoid IP agonists (MRE-269 and BMY 45778) were selected on their physi- Pharmacovigilance Database cochemical properties. In experiment 1 drugs were delivered by cathodal or ano- = I Lacroix, C Soussan, G Portolan, JL Montastruc, The French Association of dal iontophoresis onto the hindquarters of anaesthesized rats (n 8 for each Regional Pharmacovigilance Centres CHU de Toulouse, Universite de Toulouse, drug) and skin blood flow was quantified using laser Doppler imaging. Experi- Inserm 1027, Toulouse, France ment 2 aimed at comparing the effect of three different concentrations of the can- Introduction: It is well established that most of drugs were excreted into breast didates selected in experiment 1; tolerance was also assessed by recording blood milk and thus also available to the infant. The effects on the infant, of many pressure continuously, and by histopathologic examination of full-thickness skin drugs taken by the mother are unknown. biopsies. We finally aimed at optimizing drug delivery by comparing continous Objective: To describe Adverse Drug Reactions (ADRs) registered in the French vs. sequential iontophoresis protocols (experiment 3). Pharmacovigilance Database. Results: Anodal iontophoresis of A-350619 (7.54 mM) induced a significant and = Method: All spontaneous reports of ADRs in breastfed infants recorded by the sustained increase in cutaneous blood flow (P 0.008, vs. NaCl). The other 31 French Regional Pharmacovigilance centres in the National Pharmacovigi- drugs we tested exhibited poor or no effect. We observed a concentration-depen- < lance database were investigated. dent vasodilation when delivering A-350619 through iontophoresis (P 0.001, Results: Between January 1985 and June 2011, 174 ADRs in breastfed infant Jonckheere-Terpstra trend test). Finally, tolerance was good for A-350619, with were notified to the French network of PharmacoVigilance. Median age was no evidence of skin damage, no significant effect on systemic blood pressure and 49.0 Æ 66.7 days. Mean weight (3863.9 Æ1326.9 g) and length (49.4 no cutaneous resistance change. Æ3.0 cm) of infants were lower than in the French general population. The most

often reported ADRs were nervous system disorders (28.6%), followed by gastro- 26-04 intestinal disorders (20.3%) and skin and subcutaneous tissue disorders (6.5%). Pregnancy outcomes after maternal use of azathioprine: a French cohort Sixty-five (37.4%) ADRs were considered as serious. Most frequently suspected Study drugs were nervous system drugs, mainly antiepileptics, benzodiazepines and opi- Z Alamia, H Cissokob, S Ahidc, N Bernardd, A Disson-Dautrichee, L Lagarcef, oid analgesics. Drugs more often suspected in serious ADRS were dex- A Toutaing, Y Cherrahh, AP Jonville-Berai aDepartment of Pharmacology and tropropoxyphene (respiratory distress, apnea…), ketoprofene (renal and digestive Regional Pharmacovigilance Center, Tours and Department of pharmacology, Medical adverse effects…), lamotrigine, hydroxyzine and clonazepam. and Pharmacy School, Mohamed The first University, Oujda, Morocco, Tours, France; Conclusion: Some drug classes such as opioids and antiepileptics drugs, NSAIDs bDepartment of Pharmacology and Regional Pharmacovigilance Center, Tours, France; and benzodiazepines, which produced adverse effects in the infant, should be cPharmacoepidemiology and Pharmacoeconomics Research Team, Medical and used, when necessary, with great care pharmacy School, Mohammed V – Souissi University, Rabat, Morocco; dRegional Keywords: Adverse Drug Reaction, Pharmacovigilance, Breastfeeding, drugs Pharmacovigilance Center, CHU Lyon, Lyon, France; eRegional Pharmacovigilance Center, CHU Dijon, Dijon, France; fRegional Pharmacovigilance Center, CHU Angers, Angers, France; gDepartment of Medical Genetics, CHRU Tours, Tours, France; 26-02 hPharmacoepidemiology and Pharmacoeconomics Research Team, Medical and i Veinotonics in pregnancy: a comparative study in the EFEMERIS pharmacy School, Mohammed V – Souissi University, Rabat, Morocco; Department of database Pharmacology and Regional Pharmacovigilance Center, CHRU Tours, France, Tours I Lacroixa, AB Beaua, C Hurault-Delaruea, JL Montastruca, C Damase- cedex, France Michela aCHU de Toulouse, Universite de Toulouse, Inserm 1027, Toulouse, France Although azathioprine is considered relatively safe during pregnancy, it remains Introduction: There is few published data about possible effects of veinotonics in listed as a category « D » drug by the FDA. pregnant women. However, many French women use these medications during Objectives: Our main purpose was to examine the rate of malformations and their pregnancy. spontaneous abortions in women treated with azathioprine during the first tri- Objective: The present study investigates potential adverse drug reactions of mester of pregnancy. Prematurity and low birth weight in infants exposed to aza- veinotonics in pregnancy. thioprine during the third trimester were also analysed. Method: EFEMERIS is a database including prescribed and dispensed reimbursed Study design: In this multicenter prospective study, pregnant women exposed to drugs during pregnancy (data from Caisse Primaire d’Assurance Maladie of azathioprine during the 1st trimester were included after request for risk assess- Haute-Garonne) and outcomes (data from Maternal and Infant Protection Service ment. Each patient was matched with one disease-paired control that used other and from Antenatal diagnostic Centre). Women who delivered from July 1st immunosuppressant than azathioprine for similar indications (excepting myco- 2004 to September 30th 2008 in Haute-Garonne (time period when veinotonics phenolate mofetil). The primary endpoint was to evaluate the incidence of major were still reimbursed) and were registered in the French Health Insurance Service malformations. have been included into the EFEMERIS database. We compared pregnancy out- Results: One hundred and twenty four pregnant women exposed to azathioprine comes and newborn health between women exposed to veinotonics during preg- during the 1st trimester of pregnancy were included. The rates of spontaneous nancy and unexposed women. Malformations were classified according to abortions did not differ between the 2 groups (4.9 vs. 3.6%, P = 0.7). The rate of Eurocat classification. all birth defects was 7.2% in the azathioprine group vs. 5.4% in the other immu- Results: 9,671 (25.7%) newborns exposed in utero to veinotonics were compared nosuppressant group [RR = 1.36 (0.4–4.2)]. The rate of major malformations with 27 982 controls (non exposed newborns). The most widely used veinotonics was slightly higher in the azathioprine group (5/97 or 5.2%) compared to dis- were hesperidin, diosmin and troxerutin. The mean age of the mothers was ease-matched controls (2/110 or 1.8%) but did not significantly differ between 31.2 Æ 4.8 years in the exposed group and 30.0 Æ 5.1 in the control group groups RR 2.96 [0.5–15.6]. Two of the 5 major birth defects from the azathio- (P < 10À4). The mean number of drugs taken during pregnancy was higher in prine group were observed in women also exposed to tacrolimus for organ trans- the exposed group (13.4 Æ 8 vs. 9.4 Æ 7; P < 10À4). Pregnancies led to 98.2% plantation. Birth weight (3225 g vs. 3279 g), gestational age at delivery (37.7 vs. 93.4% of live-births, 0.2% vs. 0.2% of postnatal deaths and 1.6% vs. 6.4% of vs. 38 w), and preterm deliveries (22.6% vs. 21.4%) were comparable between pregnancy termination (miscarriage, ectopic pregnancy, medical termination, both groups. intra uterine death) in exposed and non exposed groups respectively. In the Conclusions: Azathioprine exposure during organogenesis does not appear to group of newborns whose mother had a prescription of veinotonics during organ- increase the risk of major malformations but larger studies are needed to confirm ogenesis, 42 out of 1,360 (3.1%) had a malformation vs. 790 (3.0%) in the con- this observation. Exposure throughout pregnancy was not associated with an trol group (P = 0.6). The rate of neonatal pathologies was higher in the control increased risk of low birth weight or preterm delivery. group (5.8 vs. 6.4, P = 0.04). Keywords: Azathioprine, pregnancy, prospective study Conclusion: We found no increased risk of adverse pregnancy outcome including neonatal pathology and congenital malformation among women exposed to veinotonics compared with unexposed pregnant women. 26-05 Keywords: EFEMERIS, veinotonics, pregnancy, pharmacoepidemiology Pregnancy outcome in women exposed to aripiprazole during the embryonic period: a prospective multicentric cohort F Belleta, MN Beyensa, N Bernardb, D Beghinc, E Elefantc, T Vialb aCentre Regional 26-03 de Pharmacovigilance, CHU de Saint-Etienne, Saint-Etienne; bCentre Regional de c Pregnancy outcome in women exposed to dopamine agonists during Pharmacovigilance, Hospices Civils de Lyon, Lyon, France; Centre de Reference sur les pregnancy: a study in EFEMERIS database Agents Teratogenes, Hôpital Armand Trousseau, Paris, France C Hurault-Delaruea, JL Monstastruca, AB Beaua, I Lacroixa, C Damase- Objectives: Aripiprazole, an atypical antipsychotic, is not recommended during Michela aCHU Toulouse, Universite Paul Sabatier de Toulouse, INSERM, Toulouse, pregnancy because of teratogenicity suggested in animal studies and the paucity France of clinical data. The main objective of our study was to evaluate whether aripip- Objective: Dopamine agonist drugs can be prescribed for several indications like razole exposure during the embryonic period was associated with an increased hyperprolactinemia or Parkinson’s disease. Prevalence of these diseases is low risk of major malformations. Secondary objectives were to evaluate the risks of during pregnancy. Thus, very little is known on the possible effect of dopaminer- miscarriage, prematurity, fetal growth retardation and maternal complications gic agonists on embryo-fetal development. The aim was to describe pregnancy and to describe possible neonatal adverse effects. outcomes in women exposed to prescription of dopamine agonists in EFEMERIS, a Methods: We conducted a multicenter cohort study using data prospectively col- cohort of French pregnant women. lected by the French Regional Pharmacovigilance Centres participing to the Te- Method: An ‘exposed-non exposed’ study was conducted in EFEMERIS cohort rappel program and the Centre de Reference sur les Agents Teratogenes (CRAT) (database including prescribed and dispensed drugs during pregnancy and out- between 2004 and 2011. ‘Exposed patients’ were pregnant women exposed to comes). The present study concerns the 57 408 mother-outcomes pairs aripiprazole during embryogenesis, i.e. 4–10 weeks after the last menstrual per- included in EFEMERIS database between 2004 and 2010. Women who received iod (exclusion of patients co-exposed to known teratogenic agent). ‘Unexposed at least one dispensation of a dopamine agonist drug during pregnancy were patients’ were pregnant women without exposure or exposed to non-teratogenic considered as exposed to dopamine agonist drugs and classified in the agent during embryonic period. Each ‘exposed patient’ was matched with two ‘exposed group’. They were individually matched to 2 ‘unexposed’ women. We ‘unexposed patients’ for age (Æ2 years) and gestational age at call (Æ2 weeks). compared adverse foetal outcomes in the two groups using conditional logistic Results: Eighty-six patients were included in the exposed group and 172 in the regressions. unexposed group. Compared to unexposed patients, exposure to aripiprazole was Results: One hundred and eighty-three women (0.3%) had a dispensation of at not associated with a significant increased risk of major malformations least one dopamine agonist during pregnancy. Bromocriptine was the most-pre- (OR = 2.30; 95%CI= 0.32–16.69) or miscarriage (OR = 1.66; 95%CI=0.63– scribed drug (more than half of the exposed women) followed by cabergoline 4.38) or gestational diabetes (OR = 1.15; 95%CI=0.33–4.04); but was associated (20%) and quinagolide (10%). Most of the indications (67%) were for hyperprol- with a significant increased risk of prematurity (OR = 2.57; 95%CI=1.06–6.27) actinemia. 75% of dopamine agonist prescriptions were performed during the first and fetal growth retardation (OR = 2.97; 95%CI=1.23–7.16). Among the 19 trimester of pregnancy. Prescriptions strongly decreased during the second tri- newborns exposed to aripiprazole near delivery, there were one case of with- mester (8.8%). There was no difference between the two groups concerning preg- drawal syndrome with pulmonary hypertension and respiratory distress and one nancy history and demographic data. Women exposed to a dopamine agonist case of amniotic fluid aspiration pneumonia. received more prescriptions of other active substances during pregnancy than the Conclusion: Our study suggests that aripiprazole is not associated with a major unexposed group. After adjustment for potential confounders, the risks of preg- teratogenic risk. These preliminary results support the reassuring limited pub- nancy termination and preterm birth were significantly increased after exposure lished data, but must be confirmed by more powerful studies. to dopamine agonist drug during pregnancy with respectively a prevalence odds Keywords: aripiprazole, pregnancy, malformations ratio of 3.7 (95% CI 1.8, 7.4) and 2.7 (95% CI 1.04, 7.0). The prevalence of birth defect and low birthweight was not statistically different between the two groups. No difference in psychomotor development at 9 and 24 months of babies was observed. Discussion: The results of this study suggest that fetal exposure to dopamine agonist drugs may increase the risk of adverse fetal outcomes. Due to limited enrolment, further studies are needed for better evaluation of these risks. Keywords: Pregnancy, prescriptions, dopamine agonist, outcome, pharmacoepidemiology

26-06 ischemia during exercise should be segmental (i.e.: able to differentiate the blood First trimester exposure to domperidone: a comparative prospective flow impairment in different muscles of the same limb) reliable and non invasive. study Arterial imaging such as resting ultrasounds or arteriography cannot account for J Cottina, D Beghinb, AP Jonville-Berac, M Boyerd, M Zenute, C Damase-Michelf, the potential blood supply during exercise because of the collateral circulation. E Elefantb, J Descotesa, T Viala aCentre Regional de Pharmacovigilance-Centre Measurements of arterial pressure at rest or after exercise provide some informa- antipoison, Hospices Civils de Lyon, Lyon, France; bCentre de Reference sur les Agents tion on the post exercise residual pressure while blood flow slowly decreases in Teratogenes, Hôpital Armand Trousseau, Paris, France; cCentre Regional de the post exercise period. Nevertheless, the technique is blind to what occurs dur- Pharmacovigilance, CHRU de Tours, Tours, France; dCentre Regional de ing exercise. Thallium scintigraphy has been shown to be a sensitive approach to Pharmacovigilance, Assistance Publique-Hôpitaux de Marseille, Marseille, France; the problem of exercise-induced ischemia, but provide only relative changes from eCentre Regional de Pharmacovigilance-CHU de Clermont Ferrand, Equipe EA 4681 reference areas. Nuclear magnetic resonance (NMR) imaging and spectroscopy PEPRADE-Universite d’Auvergne, Clermont Ferrand, France; fCentre Regional de have been used before and after stress tests to measure metabolism but, to date, Pharmacovigilance, CHU de Toulouse, Toulouse, France are not fast enough to study metabolism during muscle contractions. Near infra Background: Domperidone is a prokinetic and antidopaminergic drug used to red spectroscopy (NIRS) and transcutaneous oxygen pressure measurements treat nausea and/or vomiting. Reproductive toxicity studies have found embryo- (tcpO2) are theoretically easier for clinical use but remain surface techniques. logical abnormalities in only one species and at maternally toxic doses. As only Future developments are required to quantify tissue ischemia. Indeed, studies very few clinical data are available, we conducted a prospective comparative attempting to show the biological effects of exercise induced ischemia generally study to determine whether domperidone exposure during the first trimester of report only weak correlations with the quantification of ischemia. Further, stud- pregnancy is associated with an increased risk of major malformations. ies analysing tissue consequences of muscle ischemia show a wide heterogeneity Method: Prospective data collected by 12 French pharmacovigilance centers and of results that might be explained by differences in the severity, duration and dif- the Centre de Reference sur les Agents Teratogenes (CRAT) from 1995 to 2011 fusion of ischemia. The presentation will review the advantage and limits of were analysed. Patients were included if exposed to domperidone within 4– available tools and some perspective for future studies of the patho-physiology of 12 weeks after the last menstrual period and not exposed to any other antiemetic exercise-induced ischemia. drug during embryogenesis. Each exposed woman was matched for age (Æ2 years) and gestational age at the time of first request (Æ1 week) with two healthy women counselled after a non-teratogenic exposure during the same period. Results: Data on 121 exposed pregnancies and 244 controls were collected. Maternal age (32 Æ 5.4 years vs. 31.4 Æ 5.2, respectively) and gestational age Ischemia and Mitohormesis: therapeutic promises? at inclusion (11.6 Æ 6.6 weeks vs. 11.5 Æ 6.5 weeks) did not differ between B Genya, A Meyera, A Schlagowskia, F Singha, A Lejaya, A Charlesa, F Piquarda, groups. There were 109 live births (90.1%) among the exposed women compared J Zolla aInstitut de Physiologie, FacultedeMedecine, 11 rue Humann, Strasbourg, to 218 (89.5%, 4 sets of twins) in the controls. The rate of spontaneous abor- France tions (7.6% vs. 6%) and elective terminations of pregnancy (4.1% vs. 3.3%) did Vascular surgery is commonly carried out for lower limb ischemia in high-risk not significantly differ between the exposed and control groups. After exclusion of surgical patients with advanced cardiovascular disease. Skeletal muscle ischemia- one case of Down syndrome in the exposed group, the rate of major malforma- reperfusion (IR) injury largely participates in perioperative and long-term morbid- tions among newborns and examinable fetuses was 2/109 (1.8%) in the exposed ity, likely through increased oxidative stress. Interestingly, the mitochondrion is a group and 6/222 (2.7%) in the control group (relative risk: 0.67 CI: 0.14–3.31). central player in cell survival, being implicated in ATP production, in ROS gener- Conclusion: Despite limitations due to the sample size, the results of this pro- ation and in apoptosis. Protecting mitochondria will protect the organ involved spective study suggest that exposure to domperidone during the first trimester of directly in IR but will also protect remote organs implicated in the multi-organ pregnancy is not associated with an increased risk of malformations. failure syndrome. Therefore, reducing mitochondrial dysfunctions become a new Keywords: domperidone, pregnancy, prospective study challenge. This can be obtained by reducing the amount or ROS produced and/or by increasing anti-oxidant defenses. Mitohormesis belongs to the second option. Specifically, hormesis is an adaptive response to cellular stresses that contributes to cell protection. This corresponds to the general concept of ‘stress-response hor- ISCHEMIE TISSULAIRE PERIPH ERIQUE mesis’, considering that stress might induce stress-protective mechanisms. Mito- chondrial hormesis or mitohormesis can be defined as increased mitochondrial biogenesis, triggered by low doses of mitochondrial ROS generation. Such defense Pressure-induced vasodilation: the missing link in Diabetic foot mechanism has been described in several settings and might participate in life occurrence? a a span extension. Interestingly, and given rise to difficulties when clinical applica- D Sigaudo-Roussel UMR 5305 CNRS. Biologie Tissulaire & Ingenierie tions to be performed, a same amount of ROS might be protective or deleterious, Therapeutique, Lyon, France depending on target tissue characteristics. Thus, an opposite effect of statins on A functional skin is preserved from skin lesions in response to mechanical and cardiac and skeletal muscle has been reported, that involves ROS and peroxisome thermal stimuli but this feature is lost during aging, diabetes and under stress. proliferator-activated receptor gamma co-activator (PGC-1) family, a major indu- Despite numerous studies of prevention and treatment of diabetic foot (DF) made cer of mitochondrial biogenesis. over the last two decades, the rate of lower limb amputation remains high Concerning lower limb ischemia, several argument strongly support that mito- among diabetic patients (15 times) compared to non-diabetic patients. 15% of hormesis might be a promising therapeutic approach. diabetics have a DF in their lives associated with a risk of amputation and mor- First, data demonstrate clearly that increased oxidative stress is involved in the tality two times greater than that of a diabetic population without DF. Predict deleterious effects of IR and that reducing ROS production reduces mitochondrial the occurrence of DF is limited, and so far, only the occurrence of an injury trig- dysfunction. Ischemic post-conditioning and delayed reperfusion allows thus to gers a process of evaluation and treatment. reduce the amount of ROS reaching ischemic organ during reperfusion. Our team is behind the discovery of the Pressure-Induced Vasodilation (PIV) first Second, application of repeated cycles of reperfusion before sustained ischemia, i.e observed in healthy subjects after local application of a gradual pressure on the ischemic preconditioning also reduces mitochondrial dysfunction likely through skin leading to cutaneous vasodilation at the application of pressure. This gain in muscle anti-oxidant capacity enhancement and subsequent decrease in ROS. blood flow delays the onset of ischemia. The development of PIV has been Finally, skeletal muscle characterized by increased mitochondrial number and observed in rodents and allowing to identify the underlying mechanisms. Over level of oxidative capacity shows higher antioxidant capacity and is more pro- the past five years we have shown that in pathological conditions (eg diabetes, tected against IR than more ‘glycolytic fibres’. aging…), PIV was absent and was related to increase pressure ulcer risk. More- Thus, one might assume that therapeutic approaches aiming to increase mitohor- over the combination of peripheral vascular and nerve alteration makes a dia- mesis might protect against the deleterious effects of lower limb IR. Old strategies betic patient particularly susceptible to foot ulceration. We have already shown deserve to be re-evaluated. Exercise is known to increase biogenesis and should the deleterious effect of chronic neuropathy on the neuro-vascular interaction in be used preventively more widely to reduce muscle alteration in the setting of diabetic mice but we also showed an alteration of the cutaneous microcirculation major vascular surgery. Similarly, and more surprisingly, simple massage might foregoing neuropathy in diabetes. Thus, individuals lacking a normal PIV play a similar protective role. Local, or preferentially, remote ischemic precondi- response show an early decrease of cutaneous blood flow to the application of tioning has already been shown to be protective in humans. Stem cells or factors very low pressures reflecting a vascular fragility of the skin that increases the risk stimulating mitochondrial biogenesis still need further evaluations. of ulceration. Recent lab experiments showed the relationship between presence of PIV and absence of pressure-induced skin wound and vice versa, absence of PIV and appearance of pressure-induced skin wound in diabetic animals. We identified a cutaneous mechanosensor that is involved in initiating both the vasodilation under low pressure and reactive hyperemia after high pressure compres- Arterial ischemia: interaction between large arteries, resistance arteries sion. Absence of PIV informs on the failure of the skin capacity to fight against and the microcirculation post-compression ischemia that could weaken the tissues and could promote the D Henriona aINSERM U1083 - CNRS UMR 6214, Laboratoire de Biologie development of skin lesions Neurovasculaire et Mitochondriale Integree, Angers, France Chronic hypertension and diabetes mellitus induce macrovascular and microvascular pathophysiological modifications. Stiffening of large arteries increases central systolic and pulse pressure, which both contribute to raise left ventricular afterload and reduce coronary perfusion. On the other hand, resistance arteries Quantification of exercise ischemia in patients with arterial claudication: undergo remodelling and capillary rarefaction which both increase peripheral clinical tools and perspective in physiopathology a b,c b,c d a a resistance. Consequently, hypertension develops and amplifies the detrimental P Abraham , M Feuilloy , D Schang , D Henrion , G Leftheriotis Explorations haemodynamic effects of arterial stiffening. This vicious circle leads progressively fonctionnelles vasculaires, centre regional de medecine du Sport, Centre hospitalier to target organs impairment. Hypertension and diabetes are the major risk factors universitaire, LUNAM, BNMI, UMR CNRS 6214-INSERM 1083, Universite b involved in the development of large arteries atherosclerosis. This later in major d’Angers, Angers, France; ESEO, 10 Bd Jeanneteau, CS 90717, 49107 Angers leg artery reduces blood supply, which progressively leads to limb ischemia. Pres- Cedex 2, France; cLUNAM, LAUM – UMR CNRS 6613, Avenue Olivier Messiaen, d sure unloading and ischemia in distal arteries produces important changes in 72085 Le Mans Cedex 9, Angers, France; LUNAM, BNMI, UMR CNRS 6214- microvascular function. Indeed, resistance arteries undergo significant wall thin- INSERM 1083, Universite d’Angers, Angers, France ning and changes in contractile function. Control of large artery diameter by Quantification of tissue ischemia during exercise remains a puzzling issue in small arteries through flow-mediated vasodilation is also deeply impaired. Angio- patients with arterial claudication. The ideal technique to quantify muscle genesis is stimulated, which can induce the formation of collateral feeding blood

vessels. However, angiogenesis may also take part to atherosclerotic plaques SESSION MIXTE IMAGERIE DU PETIT ANIMAL destabilization, which then induces further deterioration in blood flow supply. Surgical bypass grafting aiming to restore blood supply to distal tissues generates Imaging modalities in preclinical cerebral stroke research a sudden increase of pressure in a weakened resistance vasculature, leading to B Guilleta aFaculte de Pharmacie, laboratoire pharmacodynamie, 27 bd Jean Moulin, uncontrolled changes in capillary hydrostatic pressure, extravasation of fluid, and Marseille, France tissue edema. The major unresolved question is why the physiological mecha- Ischemic stroke is a major cause of morbidity and mortality worldwide. A stroke nisms that regulate vascular structure and function ultimately break down, lead- is an acute neurologic injury in which the blood supply to a part of the brain is ing to circulatory failure within the distal limb. interrupted. That is, stroke involves sudden loss of neuronal function due to disturbance in cerebral perfusion, commonly arterial. The part of the brain with disturbed perfusion no longer receives adequate oxygen. This initates the ischemic cascade that makes brain cells die or be seriously damaged, impairing local brain function. Most strokes are due to obstruction of an artery in the brain by a blood Cardiovascular impact of soluble epoxide hydrolase inhibition in clot and less than 10 % of patients are eligible for thrombolysis, the sole available experimental type 2 diabetes therapy. C Rochea, M Besniera, N Haroukia, D Coquerela, A Guereta, L Nicola, b c a a c a It is therefore crucial to develop neuroprotective and/or neurorestorative thera- C Morisseau , V Richard , P Mulder , A Ouvrard-Pascaud , J Bellien Inserm pies. These developments need the use of efficient tools to accurately evaluated U1096, FacultedeMedecine - Pharmacie, Bvd Gambetta, Rouen, France; bUniversite c tissue insult and regeneration. Among them, imaging, and particularly modalities de Californie, Davis, CA, USA; Inserm U1096 - CHU de Rouen, Rouen, France using radionuclide, is gaining widespread use in acute stroke model experimenta- Objective: This study investigates the impact of the pharmacological inhibition tions for the quantification of physiopathological processes at nanomolar level. of soluble epoxide hydrolase (sEH), which increases epoxyeicosatrienoic acid The two nuclear imaging modalities used are Single Photon Emission Computer- (EET) bioavailability, on the cardiovascular abnormalities associated with type 2 ized Tomography (SPECT) and Positron Emission Tomography (PET). These diabetes. = modalities allow, using specific radiotracers, to quantify longitudinally post-stroke Methods and results: FVB mice were subjected to a control chow diet (n 30) physiopathological and regeneration processes. For example, during the early or an high-fat diet (HFD) for 16 weeks. HFD mice received during the last steps of ischemic stroke on preclinical model, 99mTc-HMPAO and 18F-Mis- 8 weeks the sEH inhibitor t-AUCB (10 mg/L in drinking water, n = 36), gliben- = = onidazol imaging respectively evaluate the transient reduction in cerebral blood clamide (80 mg/L, n 30) or were not treated (n 28). As compared with con- flow and level of tissue hypoxia. Consecutively, blood brain barrier disruption, trol mice, non-treated HFD mice had increased fasting plasma glycemia and involved in brain oedema and inflammatory cells recruitment, can be quantified insulinemia, impaired glucose and insulin tolerance, without change in blood using high hydrophilic radiotracers such as 99mTc-DTPA. Using 18Fluorodeoxy- pressure. HFD mice exhibited decreased myocardial perfusion (MRI) and cardiac glucose (18FDG) and 99mTc-Anexin nuclear modalities allow to quantify respec- diastolic dysfunction, illustrated by the decrease in the E/A ratio (echocardiogra- tively the ischemic insult on cells viabilities or apoptosis activation. Then, at phy) and the increase in the slope of the end-diastolic pressure-volume relation delayed time points, microgliale activation, angiogenesis, glial scaring and neuro- (invasive hemodynamics). Moreover, HFD mice had a profound impairment of genesis can also be analysed. coronary endothelium-dependent relaxations to acetylcholine (myography) due to In conclusion, nuclear imaging modalities represent a helpful tool in preclinical a decrease in both NO and EET availability, as demonstrated by the decreased research aimed on cerebral stroke therapies. Initially inaccessible for scientist, the sensitivity to inhibitors of NO-synthase and of CYP450 epoxygenases, the EET- increase in the number of PET and SPECT platform now allows a wider use of synthesizing enzymes, without change in endothelium-independent relaxations to these technologies accessible to the entire scientific community. sodium nitroprusside. Fasting glycemia was reduced by t-AUCB and glibenclamide, but t-AUCB also decreased insulinemia and improved glucose tolerance. Moreover, only t-AUCB prevented the development of diastolic dysfunction, and normalized coronary endothelial function, by restoring both NO and EET pathways. Neither glibenclamide nor t-AUCB improved myocardial perfusion. Multicolor fiber-based endomicroscopy: Redefining the understanding of Conclusion: These results demonstrate that, independently from its glucose-low- in vivo biology ering effect, the inhibition of sEH represents a promising pharmacological target H Ghrabia aMauna Kea Technologies, 9, rue d’Enghien, Paris, France to prevent the endothelial and cardiac dysfunction associated with type 2 diabe- Since its inception in the field of endomicroscopy, several years ago, fiber based, tes and thus, may be useful to improve cardiovascular prognosis in diabetic or probe based, confocal endomicroscopy (pCLE) has extensively proven the bene- patients. fit of in situ & realtime examination of living tissues at the microscopic scale. Keywords: Insulin-resistance, type 2 diabetes, endothelial function, cardiac func- The constant efforts devoted by Mauna Kea Technologies at increasing image tion, epoxyeicosatrienoic acids, soluble epoxide hydrolase quality, reducing invasiveness or improving system’s ergonomics have turned pCLE not only into an irreplaceable research and diagnosis instrument, but also an accurate decison making tool in the clinics, either in GI endoscopy or pulmonology. A straightforward extension of pCLE approach was thus multicolor endomicroscopy, where simultaneous multi-laser excitation, again coupled with minimally invasive and microscopic resolution thin and flexible optics, not only brings complementary and valuable biological informations, but also paves the way to a combination of morphological and functional imaging. We will present you a new system, Cellvizio Dual Band, capable of video rate (12 fps) in vivo & in situ fluorescence imaging with a microscopic resolution (1.4 lm). In its standard configuration, the system simultaneously operates at 488 and 660 nm, where it automatically performs the necessary spectral and photometric calibrations and where it provides unambiguously co-registered images, in realtime. The main hardware and software features (laser power control, sensitivity adjustment, image fusion, data exports) will be presented as well. We will also cover a panorama of its current applications, illustrated with recent results, both in the field of preclinical and clinical imaging.