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Figure S2: Knockdown of PRDX1 in MFs results in characteristics found in CAFs and JNK activation. (A) Spontaneously immortalized MFs isolated from 8-wk-old virgin female Balb/c mice were infected with lentiviruses expressing 4 different shPRDX1 targeting shRNAs and analysis by immunoblotting for PRDX1 expression. (B) α-SMA (red) staining and light microscopy of Balb/c MFs expressing EV or shPRDX1. Scale bar in 1 μm. (C) Prdx1−/− and Prdx1+/+ MEFs were analyzed by immunoblotting for phosphorylation of c-jun, ATF2 and JNK. (PPTX 520 kb)

Figure S1: PRDX1-deficiency in MFs induces characteristics found in cancer-associated fibroblasts. MFs isolated from female 8-wk-old Prdx1−/− and Prdx1+/+ mice were analyzed for α-SMA (red), vimentin (red) and Hoechst nuclear stain (blue) by IF. Scale bar in 1 μm. (PPTX 2482 kb)

Objective The aim of our study was to assess the impact of psychiatric medications and concomitant risk factors on the prevalence of QTc prolongation and torsades de pointes (TdP) in hospitalized subjects. We examined the association between individual risk scores and QTc prolongation and proposed an evidence-based protocol for electrocardiogram monitoring on psychotropic medications. Method Electrocardiograms (ECGs) of subjects hospitalized over a 1-year period were analyzed for QTc prolongation, associated risk factors, and use of medications. Analysis was performed using logistic regression to identify independent predictors of QTc prolongation, and the Pearson χ test was used for risk score assessment. Results A total of 1249 ECGs of 517 subjects were included in this study. Eighty-seven subjects had QTcB intervals greater than 470 milliseconds for females and greater than 450 milliseconds for males. Twelve (2.3%) subjects had QTcB of 500 milliseconds or greater, or greater than 60 milliseconds of change from baseline. Of these subjects, only 1 case of QTc interval change was related to routine use of psychiatric medications. There were no incidents of TdP. Age, diabetes, hypokalemia, overdose, diphenhydramine, and haloperidol were significant independent predictors of QTc prolongation. Risk scores were significantly correlated with QTc prolongation (P = 0.001). Conclusion Our retrospective review study found that the occurrence of TdP and QTc prolongation was low in this subject population. QT abnormalities were associated with known risk factors, and risk scores correlated well with QTc prolongation. Providers can use the protocol proposed in this study, which incorporates risk scores and the CredibleMeds classification system to determine the need for ECG monitoring and to guide treatment.

Background: The benefits of post-operative radioactive iodine (RAI) administration have not been demonstrated in patients with low risk differentiated thyroid cancer (DTC). The objective of this randomized phase III trial is to assess in low risk DTC patients the non-inferiority of a follow-up strategy as compared to a systematic adjuvant post-operative RAI administration. Methods: ESTIMABL2 is a French multicentric randomized phase III trial in patients with low-risk DTC treated with total thyroidectomy with or without prophylactic neck lymph node dissection (pT1am N0 or Nx with a sum of the diameters of tumor lesions ≥ 10mm, pT1b N0 or Nx). Two to five months after surgery, in the absence of suspicious lateral neck lymph node on ultrasonography (US), patients were randomized either to the follow-up group (FU, no RAI administration) or to the ablation group and received post-operative RAI (1.1 GBq following rhTSH stimulation). Yearly controls under levothyroxine treatment consisted in thyroglobulin (Tg) and Tg antibodies (TgAb) determinations and neck-US. The primary objective was to assess at 3 years after randomization the non-inferiority of the proportion of patients without tumor-related event in the FU group as compared to the ablation group. Non-inferiority is demonstrated if the rate of patients without event at 3 years does not differ by more than ΔL=-5%. A tumor-related event was defined by the occurrence of subsequent treatment (RAI administration or surgery) for abnormal RAI uptake on the post-therapeutic WBS or by elevated Tg or TgAb levels and/or abnormal neck US during controls. Tg levels on levothyroxine treatment were considered elevated if > 2ng/mL in the FU group and > 1ng/mL in the ablation group. TgAb were considered elevated if > the upper limit range with an increase above 50% on 2 consecutive determinations performed 6 months apart. Results: 776 low-risk DTC patients were included between 2013 and 2017 in 35 French centers within the TUTHYREF network; 83% females, mean age: 52 years, papillary TC: 96%, pT1bNx: 43.6%, pT1bN0: 37.5%, pT1amNx: 12.6%, pT1amN0: 6.3%. Among the 729 patients evaluable at 3 years after randomization, tumor-related events occurred in 18/367 patients (4.9% IC95%=[2.9; 7.6]) in the FU group and in 15/362 patients (4.1% IC95%=[2.3; 6.7]) in the ablation group. Thus, 95.1% of patients in the FU group had no event at 3 years and this percentage is not inferior from the 95.9% of patients observed in the ablation group (difference = -0.8% [95% CI:-3.3%; 1.8%]. The number of subsequent surgery and/or RAI administration was 6 (1.6% IC95%=[0.6; 3.5]) in the FU group and 9 (2.5% IC95%=[1.1; 4.7]) in the ablation group. Conclusion: this phase III trial demonstrates the non-inferiority of a follow-up strategy compared to a systematic adjuvant post-operative administration of RAI (1.1GBq following rhTSH) in low risk DTC patients (PHRC 2012; NCT01837745).

Background Chronic kidney disease (CKD) defined as a glomerular filtration rate (GFR) Methods Four equations were used to calculate GFR in asymptomatic T2D patients consulting between 2007 and 2016 in our establishment for a CV assessment. Cox proportional hazard ratio was used to build and compare prediction models for each equation. Results Among 796 asymptomatic T2D patients, the prevalence of CKD was 18% (MDRD and CKD-EPI), 13% (CG-SC) and 11% (CG). The 5-year incidence of CV events was 8% (n = 63). After adjustment on covariables, CKD was associated with CV events when defined by MDRD (HRa = 2,10[1,17-3,78]) and CKD-EPI (HRa = 1,95[1,07-3,52]) but not for the other 2 formulas. Furthermore, only the prediction models including MDRD and CKD-EPI (with equal performance) provided significant information to the model without GFR (P Conclusion In this asymptomatic T2D population, we have found that MDRD and CKD-EPI equations can equally predict the cardiovascular events occurrence.

International audience; AIM: The objective of this study was to investigate the involvement of the motor fibres of the corpus callosum after unilateral neonatal arterial ischemic stroke (NAIS) of the middle cerebral artery territory and the relationship to both ipsilesional and contralesional hand function.METHOD: Using high-resolution structural magnetic resonance imaging (MRI), functional MRI, and magnetic resonance diffusion-tractography, we compared the midsagittal area of the motor part of the corpus callosum (defined by the fibres connecting the precentral gyri) between 33 7-year-old children after unilateral NAIS and 31 typically developing 7-year-old children. Hand motor performance was assessed by the box and blocks test.RESULTS: Children after NAIS showed on average significantly smaller motor corpus callosum area compared to typically developing children (p|t|)=0.034) and ipsilesional hand motor performance (Pr(>|t|)=0.006) after controlling for lesion volume and sex. In a post-hoc analysis the additional contribution of corticospinal tract damage was evaluated.INTERPRETATION: Compared to typically developing children, children after NAIS exhibited a smaller motor part of their corpus callosum associated with reduced contralesional but also ipsilesional manual dexterity. These results indicate that the affection of transcallosal motor fibres in unilateral NAIS might be of functional relevance and an important part of the involved structural network that should be elucidated in further studies.TRIAL REGISTRATION: ClinicalTrials.gov NCT02511249.© 2017 Mac Keith Press.

Introduction L’hirsutisme de la femme menopausee ne doit pas etre neglige. A cote des causes tumorales, penser a l’hyperthecose. Cas clinique Me E. 65 ans, menopausee depuis 10 ans presente un hirsutisme clinique franc, invalidant et en progression. Elle presente aussi un surpoids androide et une intolerance au glucose. La testosteronemie initiale est a 1,98 ng/ml, majoree a 2,58 ng/ml faisant craindre une tumeur du fait des taux eleves. Les dosages de SDHA, Delta 4, 17-OHP sont normaux. FSH (45 mU/l), LH (38 mUI/l) et Estrone 90 ng/ml ( Discussion La physiopathologie de l’hyperthecose ovarienne repose sur l’hyperstimulation du stroma ovarien. Nous evoquons le lien avec le SOPK, d’une part du fait de l’hyperactivite des cellules thecales et d’autre part en raison de l’effet du systeme IGFs et insulinique sur la luteinisation des cellules du stroma communes aux deux entites ovariennes. Conclusion Des liens avec le syndrome metabolique sont etablis, ils pourraient expliquer la survenue de l’hyperthecose en post-menopause.

Cardiac metastasis from renal cell carcinoma (mRCC) is very rare. An understanding of the natural history of the impact of mRCC tumors to the left ventricle is incomplete. We report a case of mRCC to the left ventricular (LV) apex with a review of literature. 67 year old man with a headache was

Flecainide toxicity is a complication of antiarrhythmic therapy with high mortality rate. Rapid recognition and appropriate therapy are important in managing complications of flecainide therapy. We report a case of flecainide toxicity with a literature review. 57 year old man with HIV, ESRD on

Aim To disentangle the respective impacts of manual dexterity and cerebral palsy (CP) in cognitive functioning after neonatal arterial ischaemic stroke. Method The population included 60 children (21 females, 39 males) with neonatal arterial ischaemic stroke but not epilepsy. The presence of CP was assessed clinically at the age of 7 years and 2 months (range 6y 11mo-7y 8mo) using the definition of the Surveillance of CP in Europe network. Standardized tests (Nine-Hole Peg Test and Box and Blocks Test) were used to quantify manual (finger and hand respectively) dexterity. General cognitive functioning was evaluated with the Wechsler Intelligence Scale for Children, Fourth Edition. Simple and multiple linear regression models were performed while controlling for socio-economic status, lesion side, and sex. Results Fifteen children were diagnosed with CP. In simple regression models, both manual dexterity and CP were associated with cognitive functioning (β=0.41 [p=0.002] and β=0.31 [p=0.019] respectively). However, in multiple regression models, manual dexterity was the only associated variable of cognitive functioning, whether or not a child had CP (β=0.35; p=0.007). This result was reproduced in models with other covariables (β=0.31; p=0.017). Interpretation As observed in typically developing children, manual dexterity is related to cognitive functioning in children having suffered a focal brain insult during the neonatal period. What this paper adds Manual dexterity predicts cognitive functioning after neonatal arterial ischaemic stroke. Correlations between manual dexterity and cognitive functioning occur irrespective of sex, lesion side, presence of cerebral palsy, and socio-economic status. Residual motor ability may support cognitive functioning.

The central serotonin

The Serotonin2A (5-hydroxytryptamin, 5-HT2A) receptor is one of the numerous seven transmembrane G protein coupled receptors for serotonin (5-HT) originally described as displaying a low affinity for its endogenous ligand. It is densely expressed in the cortex and the hippocampus of rodents, primates and humans brain. A role of 5-HT2A receptors in learning and memory has been proposed for years. In some behavioural tasks in rodents, 5-HT2A receptors would display a constitutive activity, a spontaneous activity of the receptor occurring without the presence of the endogenous ligand and silenced by inverse agonists. Nonetheless, the demonstration of the existence of such a subtle activity in living organisms relies on specific criteria and on clear-cut pharmacological evaluation. While it has been claimed that 5-HT2A receptor constitutive activity participates in the conditioned eyeblink response in rabbits, such an activity would not be systematically observed in other models of learning and conditioning such as the conditioned avoidance response in rats. Here, we propose a thorough pharmacological analysis of the available data arguing in favour of the involvement of constitutive activity of 5-HT2A receptors, mostly in learning tasks and discuss the functional significance of such an activity.

Acute lung injury (ALI) is a severe hypoxemic respiratory insufficiency associated with lung leak, diffuse alveolar damage, inflammation, and loss of lung function. Decreased dimethylaminohydrolase (DDAH) activity and increases in asymmetric dimethylarginine (ADMA), together with exaggerated oxidative/nitrative stress, contributes to the development of ALI in mice exposed to LPS. Whether restoring DDAH function and suppressing ADMA levels can effectively ameliorate vascular hyperpermeability and lung injury in ALI is unknown, and was the focus of this study. In human lung microvascular endothelial cells, DDAH II overexpression prevented the LPS-dependent increase in ADMA, superoxide, peroxynitrite, and protein nitration. DDAH II also attenuated the endothelial barrier disruption associated with LPS exposure. Similarly, in vivo, we demonstrated that the targeted overexpression of DDAH II in the pulmonary vasculature significantly inhibited the accumulation of ADMA and the subsequent increase in oxidative/nitrative stress in the lungs of mice exposed to LPS. In addition, augmenting pulmonary DDAH II activity before LPS exposure reduced lung vascular leak and lung injury and restored lung function when DDAH activity was increased after injury. Together, these data suggest that enhancing DDAH II activity may prove a useful adjuvant therapy to treat patients with ALI.

The burden of hypoglycaemia is important, particularly in elderly type 2 diabetes (T2D) patients. Unfortunately, however, few studies are available concerning this population. GERODIAB is a prospective, multicentre, observational study that aims to describe the 5-year morbidity and mortality of 987 T2D patients aged 70 years and older. After analyzing the frequency of and factors associated with hypoglycaemia in the 6 months prior to study inclusion, it was found that hypoglycaemia was associated with retinopathy, lower levels of LDL cholesterol and altered mini-Geriatric Depression Scale (GDS) scores.

Recent studies suggest that Peroxiredoxin 1 (Prdx1), in addition to its known H₂O₂-scavenging function, mediates cell signaling through redox-specific protein-protein interactions. Our data illustrate how Prdx1 specifically coordinates p38MAPK-induced signaling through regulating p38MAPKα phosphatases in an H₂O₂ dose-dependent manner. MAPK phosphatases (MKP-1 and/or MKP-5), which are known to dephosphorylate and deactivate the senescence-inducing MAPK p38α, belong to a group of redox-sensitive phosphatases (protein tyrosine phosphatases) characterized by a low pKa cysteine in their active sites. We found that Prdx1 bound to both MKP-1 and MKP-5, but dissociated from MKP-1 when the Prdx1 peroxidatic cysteine Cys52 was over-oxidized to sulfonic acid, which in turn resulted in MKP-1 oxidation-induced oligomerization and inactivity toward p38MAPKα. Conversely, over-oxidation of Prdx1-Cys52 was enhancing in the Prdx1:MKP-5 complex with increasing amounts of H₂O₂ concentrations and correlated with a protection from oxidation-induced oligomerization and inactivation of MKP-5 so that activation toward p38MAPK was maintained. Further examination of this Prdx1-specific mechanism in a model of reactive oxygen species-induced senescence of human breast epithelial cells revealed the specific activation of MKP-5, resulting in decreased p38MAPKα activity. Taken together, our data suggest that Prdx1 orchestrates redox signaling in an H₂O₂ dose-dependent manner through the oxidation status of its peroxidatic cysteine Cys52.

The thyroid is the organ with the highest selenium content per gram of tissue because it expresses specific selenoproteins. Since the discovery of myxoedematous cretinism and thyroid destruction following selenium repletion in iodine- and selenium-deficient children, data on links between thyroid metabolism and selenium have multiplied. Although very minor amounts of selenium appear sufficient for adequate activity of deiodinases, thus limiting the impact of its potential deficiency on synthesis of thyroid hormones, selenium status appears to have an impact on the development of thyroid pathologies. The value of selenium supplementation in autoimmune thyroid disorders has been emphasized. Most authors attribute the effect of supplementation on the immune system to the regulation of the production of reactive oxygen species and their metabolites. In patients with Hashimoto's disease and in pregnant women with anti-TPO antibodies, selenium supplementation decreases anti-thyroid antibody levels and improves the ultrasound structure of the thyroid gland. Although clinical applications still need to be defined for Hashimoto's disease, they are very interesting for pregnant women given that supplementation significantly decreases the percentage of postpartum thyroiditis and definitive hypothyroidism. In Graves' disease, selenium supplementation results in euthyroidism being achieved more rapidly and appears to have a beneficial effect on mild inflammatory orbitopathy. A risk of diabetes has been reported following long-term selenium supplementation, but few data are available on the side effects associated with such supplementation and further studies are required.

Objectives To evaluate the epileptic, academic, and developmental status at age 7 years in a large population of term-born children who sustained neonatal arterial ischemic stroke (NAIS), and to assess the co-occurrence of these outcomes. Study design A cohort study including 100 term newborns with NAIS was designed. Two infants died during the neonatal period, 13 families were lost to follow-up, and 5 families declined to participate in this evaluation. Thus, 80 families completed the 7-year clinical assessment. Epileptic status, schooling, motor abilities, global intellectual functioning, spoken language, and parental opinions were recorded. Principal component analysis was applied. Results Rates of impaired language, cerebral palsy, low academic skills, active epilepsy, and global intellectual deficiency were 49%, 32%, 28%, 11%, and 8%, respectively. All were highly correlated. Eventually, 59% of children were affected by at least 1 of the aforementioned conditions. In 30% of cases, the viewpoints of health practitioners and parents did not match. Conclusion The prevalence of severe disabilities at 7 years after NAIS is low, but most children exhibit some impairment in developmental profile. Trial registration ClinicalTrials.gov ( NCT02511249 ), Programme Hospitalier de Recherche Clinique Regional (0308052), Programme Hospitalier de Recherche Clinique Interregional (1008026), and EudraCT (2010-A00329-30).

Prostate cancer (PCa) is the most frequently diagnosed malignancy in men, and the second highest contributor of male cancer related lethality. Disease mortality is due primarily to metastatic spread, highlighting the urgent need to identify factors involved in this progression. Activation of the genetic epithelial to mesenchymal transition (EMT) program is implicated as a major contributor of PCa progression. Initiation of EMT confers invasive and metastatic behavior in preclinical models and is correlated with poor clinical prognosis. Extracellular Hsp90 (eHsp90) promotes cell motility and invasion in cancer cells and metastasis in preclinical models, however, the mechanistic basis for its widespread tumorigenic function remains unclear. We have identified a novel and pivotal role for eHsp90 in driving EMT events in PCa. In support of this notion, more metastatic PCa lines exhibited increased eHsp90 expression relative to their lineage-related nonmetastatic counterparts. We demonstrate that eHsp90 promoted cell motility in an ERK and matrix metalloproteinase-2/9-dependent manner, and shifted cellular morphology toward a mesenchymal phenotype. Conversely, inhibition of eHsp90 attenuated pro-motility signaling, blocked PCa migration, and shifted cell morphology toward an epithelial phenotype. Last, we report that surface eHsp90 was found in primary PCa tumor specimens, and elevated eHsp90 expression was associated with increased levels of matrix metalloproteinase-2/9 transcripts. We conclude that eHsp90 serves as a driver of EMT events, providing a mechanistic basis for its ability to promote cancer progression and metastasis in preclinical models. Furthermore, its newly identified expression in PCa specimens, and potential regulation of pro-metastatic genes, supports a putative clinical role for eHsp90 in PCa progression.

Resume Objectifs Les recommandations de prise en charge de l’obesite proposent, en l’absence d’efficacite des conseils dietetiques, de fixer le niveau des apports energetiques conseilles a une valeur proche de la depense energetique de repos (DER). L’etude avait pour but de comparer de maniere prospective pour des patients obeses adultes les mesures de DER par calorimetrie indirecte (DERcind) (methode de reference) aux resultats obtenus avec des equations predictives ainsi qu’aux niveaux d’apport issus de l’evaluation dietetique. Methodes Chaque patient beneficiait d’une mesure de DERcind et d’une estimation de DER par les formules de Harris et Benedict, de l’OMS, de Mifflin-St Jeor, de Lazzer et de Black, et par une mesure d’impedancemetrie corporelle totale. L’evaluation dietetique determinait un niveau de regime. La confrontation des resultats a ceux de DERcind utilisait la correlation de Spearman, le test de Wilcoxon et la methode de Bland et Altman. Resultats Vingt et un patients etaient inclus, d’IMC moyen 43,9 ± 6,4 kg/m2. Les correlations etaient tres significatives entre les valeurs de DERcind et celles obtenues par les equations (0,65 Conclusions Dans la population etudiee, l’utilisation des equations predictives et l’approche dietetique exposent a un fort risque de sur- ou sous-estimation des apports energetiques, non acceptable en pratique quotidienne, ce qui souligne l’interet de la calorimetrie indirecte pour fixer le niveau calorique dans la demarche de perte de poids.

Introduction Les manifestations endocriniennes les plus frequentes de l’histiocytose langerhansienne (HL) sont dues a une atteinte hypothalamo-hypophysaire representees par le diabete insipide et l’insuffisance antehypophysaire. L’infiltration de la thyroide au cours de cette maladie est rare [1] . Nous rapportons deux cas d’atteinte thyroidienne dans le cadre d’HL. Observation M. R., 18 ans, aux antecedents de comitialite et d’hypogonadisme hypogonadotrope avec IRM hypophysaire normale, consultait pour nodule thyroidien du lobe gauche. La fonction thyroidienne, les anticorps anti-thyroperoxydase et les autres axes hypophysaires etaient normaux. L’echographie montrait une thyroide multi nodulaire heterogene de taille normale. Le nodule etait hypofixiant a la scintigraphie. La cytoponction n’avait pas ete effectuee. Une thyroidectomie subtotale a ete realisee montrant un infiltrat cellulaire a noyaux irreguliers, positif pour la proteine S100, la vimentine, et le CD1, confirmant une HL. La recherche de la mutation BRAF n’avait pas ete faite. Par ailleurs il existait des lesions erythemato-squameuses et prurigineuses du cuir chevelu et des plis de flexion positives a la biopsie. Le diagnostic d’histiocytose langerhansienne de localisations thyroidienne, hypothalamique et cutanee etait retenu. Le patient fut traite par vinblastine pendant 4 ans. L’evolution fut marquee par l’apparition d’un envahissement histiocytaire osseux mastoidien bilateral confirme par biopsie. De nouvelles cures de vinblastine mensuelles furent entreprises pendant 7 ans. Devant une rechute osseuse diffuse un traitement par leustatine fut debute. M.V., 45 ans, presentait une maladie de Basedow recidivante malgre le traitement medical. Une thyroidectomie totale fut realisee dont l’analyse histologique montrait un infiltrat inflammatoire lymphocytaire diffus et la presence de grandes cellules a noyaux reniformes, exprimant le CD1a, la proteine S100, posant le diagnostic d’HL. La recherche de mutation BRAF n’avait pas ete effectuee. Le bilan d’extension s’est avere negatif. Une simple surveillance fut entreprise. Apres 3 ans de recul, il n’etait pas constate de recidive locale ou d’extension de la maladie. Conclusion L’atteinte primitive du tissu thyroidien au cours de l’HL est rare avec moins de 100 cas rapportes dans la litterature. Les femmes adultes semblent plus touchees, avec une majorite de goitres. Quarante pour cent des patients sont en euthyroidie au moment du diagnostic. Les thyroidites de Hashimoto associees sont tres rares. La cytoponction thyroidienne est peu rentable. La decouverte d’une infiltration thyroidienne histiocytaire doit motiver la realisation d’un bilan d’extension. Une atteinte uniquement thyroidienne est de bon pronostic. En revanche, elle peut s’integrer dans une atteinte multi viscerale de pronostic plus reserve. La mutation du proto oncogene BRAF commune a l’HL et au carcinome papillaire thyroidien doit etre recherchee systematiquement. Le traitement de choix est la thyroidectomie sub-totale ou totale. La chimiotherapie ne semble pas indiquee en cas d’atteinte thyroidienne unique [2] .

Chronic pain states are characterized by long-term sensitization of spinal cord neurons that relay nociceptive information to the brain. Among the mechanisms involved, up-regulation of Cav1.2-comprising L-type calcium channel (Cav1.2-LTC) in spinal dorsal horn have a crucial role in chronic neuropathic pain. Here, we address a mechanism of translational regulation of this calcium channel. Translational regulation by microRNAs is a key factor in the expression and function of eukaryotic genomes. Because perfect matching to target sequence is not required for inhibition, theoretically, microRNAs could regulate simultaneously multiple mRNAs. We show here that a single microRNA, miR-103, simultaneously regulates the expression of the three subunits forming Cav1.2-LTC in a novel integrative regulation. This regulation is bidirectional since knocking-down or over-expressing miR-103, respectively, up- or down-regulate the level of Cav1.2-LTC translation. Functionally, we show that miR-103 knockdown in naive rats results in hypersensitivity to pain. Moreover, we demonstrate that miR-103 is down-regulated in neuropathic animals and that miR-103 intrathecal applications successfully relieve pain, identifying miR-103 as a novel possible therapeutic target in neuropathic chronic pain.

International audience; Lack of compensatory or even reduced food intake is frequently observed in weight-losing cancer patients and contributes to increased morbidity and mortality. Our previous work has shown increased transcription factor expression in the hypothalamus and ventral striatum of anorectic rats bearing small tumors. mRNA expression of molecules known to be involved in pathways regulating appetite in these structures was therefore assessed in this study. Given that pain, pro-inflammatory cytokines and metabolic hormones can modify food intake, spinal cord cellular activation patterns and plasma concentrations of cytokines and hormones were also studied. Morris hepatoma 7777 cells injected subcutaneously in Buffalo rats provoked a 10% lower body weight and 15% reduction in food intake compared to free-feeding tumor-free animals 4weeks later when the tumor represented 1-2% of body mass. No differences in spinal cord activation patterns or plasma concentration of pro-inflammatory cytokines were observed between groups. However, the changes in plasma ghrelin and leptin concentrations found in food-restricted weight-matched rats in comparison to ad libitum-fed animals did not occur in anorectic tumor-bearing animals. Real-time PCR showed that tumor-bearing rats did not display the increase in hypothalamic agouti-related peptide mRNA observed in food-restricted weight-matched animals. In addition, microarray analysis and real-time PCR revealed increased ventral striatal prostaglandin D synthase expression in food-restricted animals compared to anorectic tumor-bearing rats. These findings indicate that blunted hypothalamic AgRP mRNA expression, probably as a consequence of relatively high leptin and low ghrelin concentrations, and reduced ventral striatal prostaglandin D synthesis play a role in maintaining cancer-associated anorexia.

Introduction La neurofibromatose de type 1 (NF1), ou maladie de Von Recklinghausen, maladie genetique autosomique dominante, coexiste avec des affections endocriniennes dont la plus connue est le pheochromocytome qui survient dans 0,5 a 5 % des NF1. Quelques cas d’association avec une hyperparathyroidie primaire sont decrits dans la litterature [1] . Observation Nous rapportons l’observation d’un patient âge de 34 ans porteur d’une NF1 presentant des taches cafe au lait, neurofibromes cutanes, neurofibromes plexiformes multi-etages. Decouverte d’un processus expansif de la surrenale droite lors d’un bilan d’asthenie avec presence d’une triade de Menard, hypertension arterielle orientant vers un pheochromocytome confirme sur le bilan secretoire derives methoxyles et chromogranine A et scintigraphie a la MIBG. Simultanement decouverte d’une hypercalcemie avec un profil d’hyperparathyroidie primaire avec echographie et scintigraphie des parathyroides concordantes pour un adenome parathyroidien inferieur droit. Presence egalement d’un nodule thyroidien de 3,7 cm de grand axe TIRADS 3 (calcitonine negative). Prise en charge chirurgicale : surrenalectomie droite, lobectomie thyroidienne gauche et exerese adenome parathyroidien gauche. Normalisation a 6 mois de la symptomatologie, negativation des derives methoxyles, scanner surrenalien et bilan phosphocalcique normalises. Discussion Ce cas clinique souligne que les formes frontieres (NF1, Neoplasie multiple de type 2 [NEM2]) existent probablement. D’autres equipes ont rapporte des cas similaires d’associations cliniques [2] sans demontrer a nature genetique de ce possible variant de NEM.

Resume A l’heure actuelle, le role des agonistes dopaminergiques et somatostatinergiques dans la prise en charge des adenomes hypophysaires est bien etablie. Neanmoins, la progression des connaissances sur l’expression des recepteurs dopaminergiques et de la somatostatine au niveau des cellules hypophysaires adenomateuses et le developpement de nouveaux analogues risquent de bouleverser les modalites actuelles de prise en charge. En particulier, la mise en evidence de la co-expression de differents types ou sous-types de recepteurs au niveau des cellules adenomateuses suggere des phenomenes d’interactions fonctionnelles entre recepteurs augmentant ainsi leur activite. En parallele, de nouvelles molecules sont en developpement : nouveaux analogues de la somatostatine dont l’affinite de liaison aux differents sous-types de recepteurs est plus universelle, ou molecules chimeriques capables de se fixer aux deux categories de recepteurs, somatostatinergiques et dopaminergiques. Au sein des adenomes somatotropes, on note une correlation positive entre l’expression de l’ARNm du sous-type Sst2 et l’inhibition de la secretion de la GH par les analogues de la somatostatine. Recemment l’implication du sous-type Sst5 pour les adenomes resistants aux agonistes preferentiels de Sst2 a ete demontre. Le SOM-230, agoniste somatostatinergique presentant une affinite de liaison respectivement 25, 5 et 40 fois superieure pour Sst1, Sst3 et Sst5 que celle de l’octreotide et 2,5 fois moindre pour Sst2, pourrait permettre de controler plus de patients acromegales par le traitement medical. Par ailleurs, l’utilisation d’une molecule chimerique presentant une affinite conjointe pour les sous-types Sst2 et D2 (BIM-23A287) inhibe la secretion de GH in vitro de facon similaire aux agonistes de type Sst2 ou D2 utilises seuls ou en association mais pour des concentrations 50 fois moindre. La mise en evidence des sous-types Sst5 et D2 au niveau des adenomes corticotropes ouvre de nouvelles perspectives avec des resultats preliminaires encourageants avec le SOM-230 et entraine un regain d’interet pour la cabergoline. Au sein des adenomes non fonctionnels, l’expression des sous-types de recepteurs Sst2, Sst3 et D2 permettra peut etre d’avoir recours a des therapies combinees associant les nouveaux analogues de la somatostatine aux agonistes dopaminergiques ou encore d’utiliser la do-pastatine (BIM-23 A760, molecule chimerique D2-Sst2-Sst5). Les resultats preliminaires obtenus in vitro avec cette molecule sont d’ailleurs encourageants montrant une inhibition dose dependante des mecanismes de replication cellulaire dans 60 % des cas. Enfin, au sein des prolactinomes la mise en evidence des recepteurs de type Sst5 a fait envisager l’utilisation d’agonistes somatostatinergiques specifiques de Sst5, ou du SOM-230. Neanmoins, il semble que les adenomes resistants aux agonistes dopaminergiques n’exprimant pas D2 n’expriment pas non plus Sst5. Par contre, il semble que la dopastatine soit plus efficace que la cabergoline dans la prise en charge de ce type d’adenome. Ainsi, la progression des connaissances concernant les mecanismes impliques dans le controle des secretions et de la proliferation cellulaire hypophysaire permettra peut-etre dans l’avenir de traiter la pathologie adenomateuse hypophysaire, en particulier les macroadenomes, par des traitements pharmacologiques et de ne plus avoir recours a la chirurgie et/ou a la radiotherapie hypophysaire.

The two isoforms of p190 RhoGAP (p190A and p190B) are important regulators of RhoGTPase activity in mammalian cells. Both proteins are ubiquitously expressed, are involved in the same signalling pathways and interact with the same identified binding partners. In search of isoform functional specificity, we knocked down the expression of each p190 protein using siRNA and examined the resulting phenotypic changes in human umbilical vein endothelial cells (HUVECs). We provide evidence that p190B plays a crucial role in the regulation of MT1-MMP expression and cell-surface presentation, as well as subsequent MMP2 activation. p190B is involved in both local extracellular matrix degradation at podosomes and endothelial cell assembly into tube-like structures in Matrigel. In addition, whereas p190B knockdown does not affect podosome formation, p190A knockdown increases the number of cells showing podosome structures in HUVECs. We conclude that the two p190 RhoGAP isoforms play distinct roles in endothelial cells. In addition, our data reveal an unsuspected role for p190B in the expression of the two collaborative proteases MT1-MMP and MMP2, thereby affecting matrix remodelling and angiogenesis.

Cessation of lactation causes a massive loss of surplus lactotrophs in the rat pituitary gland. The factors and mechanisms involved in this phenomenon have not yet been elucidated. Besides its inhibitory control on prolactin secretion and lactotroph proliferation, evidence suggests that dopamine (DA) may be a proapoptotic factor for lactotrophs. We therefore tested the proapoptotic effect of DA on pituitary glands from virgin, lactating, and postlactating rats. By measuring mitochondrial membrane potential loss, caspase-3 activation, and nuclear fragmentation, we show that DA induces apoptosis specifically in lactotrophs from postlactating rats. We then determined that this effect was partly mediated by the DA transporter (DAT) rather than the D2 receptor, as corroborated by the detection of DAT expression exclusively in lactotrophs from postlactating rats. We also observed tyrosine hydroxylase (TH) expression in postlactating lactotrophs that was accompanied by an increase in DA content in the anterior pituitary gland of postlactating compared with virgin rats. Finally, we observed that cells expressing TH coexpressed DAT and cleaved caspase-3. These findings show that DA may play a role in lactotroph regression during the postlactation period by inducing apoptosis. The fact that this process requires DAT and TH expression by lactotrophs themselves suggests that it may be “autocrine” in nature.

We described previously the cDNA cloning of three functional rat histamine H3 receptor (rH3R) isoforms as well as the differential brain expression patterns of their corresponding mRNAs and signaling properties of the resulting rH3A, rH3B, and rH3C receptor isoforms (Mol Pharmacol 59:1-8). In the current report, we describe the cDNA cloning, mRNA localization in the rat central nervous system, and pharmacological characterization of three additional rH3R splice variants (rH3D, rH3E, and rH3F) that differ from the previously published isoforms in that they result from an additional alternative-splicing event. These new H3R isoforms lack the seventh transmembrane (TM) helix and contain an alternative, putatively extracellular, C terminus (6TM-rH3 isoforms). After heterologous expression in COS-7 cells, radioligand binding or functional responses upon the application of various H3R ligands could not be detected for the 6TM-rH3 isoforms. In contrast to the rH3A receptor (rH3AR), detection of the rH3D isoform using hemagglutinin antibodies revealed that the rH3D isoform remains mainly intracellular. The expression of the rH3D-F splice variants, however, modulates the cell surface expression-levels and subsequent functional responses of the 7TM H3R isoforms. Coexpression of the rH3AR and the rH3D isoforms resulted in the intracellular retention of the rH3AR and reduced rH3AR functionality. Finally, we show that in rat brain, the H3R mRNA expression levels are modulated upon treatment with the convulsant pentylenetetrazole, suggesting that the rH3R isoforms described herein thus represent a novel physiological mechanism for controlling the activity of the histaminergic system.

Introduction La chirurgie bariatrique fait partie des strategies therapeutiques d’equilibration du diabete de type 2 (DT2) chez l’obese. Le court-circuit gastrique (CCG) est considere comme gold standard. Or la gastrectomie longitudinale (GL) devient la technique la plus utilisee en France. Objectifs Comparer l’efficacite de GL versus CCG dans la remission et l’equilibration du DT2 a long terme (5 ans) au CHU de Limoges et definir des facteurs predictifs de remission. Methode et materiel Etude observationnelle, retrospective de DT2 operes par GL (32) et CCG (26) entre 2005 et 2015. La remission du DT2 est definie par une HbA1c Resultats Pas de difference pour la remission du DT2 chez patients GL versus CCG : a 1an 37 % vs 42 % (p = 0,80), a 2 ans 35 % vs 57 % (p = 0,37) et a 5 ans 0 % vs 50 % (p = 0,33). L’equilibre du DT2 est maintenu a 5 ans chez plus de la moitie des sujets operes. L’absence de complications associees et un DT2 de moins de 5 ans sont significativement correles a la remission du DT2 (OR : 9,9 ; IC95 % [1,0–92] p = 0,03 et OR : 10,6 ; IC95 % [1,9–59] p = 0,007 respectivement). La baisse de l’HbA1c apparait significativement correlee a la perte d’exces de poids (p = 0,02). Discussion Cette etude montre un effet positif de la perte de poids qui est un critere conteste et confirme des criteres classiques de remission. Conclusion CCG semble plus efficace que GL sur la remission du DT2 a court et long terme. L’analyse a 10 ans est indispensable. Les mecanismes physiopathologiques restent a explorer.

This study aims to determine the prevalence of hypertension among children with diabetes mellitus, and describe adherence to the standard of practice regarding early diagnosis and treatment of hypertension in this population. Diabetes renders higher susceptibility to cardiovascular disease in affected patients and thus early detection of hypertension can be beneficial for future quality of life. We hypothesize that a number of diabetic children with hypertension are not promptly diagnosed and treated due to the difficult process preceding diagnosis. This cross-sectional study in an out-patient clinic of a university hospital was based on a group of 263 children (both females and males, age range between 3-18) diagnosed with type 2 diabetes. At the visit, the child's weight and height were recorded, BMI was calculated, systolic and diastolic blood pressure (BP) was taken, and blood sample drawn for hemoglobin A1C (HbA1C) readings. Within the study population we have identified a group of 73 children with hypertension. Comparing the two groups: the normotensive with the hypertensive, we have observed that the normotensive patients have significantly lower average BMI (26.7), p=0.000012 and slightly reduced HbA1C (8.9%), p=0.28 levels compared to the hypertensive group: BMI (32.5) and HbA1C (9.3%), respectively. We investigated the same parameters within every age group starting from age 10, and recorded that HbA1C was only significantly different for the group of 14 year-olds (8.7%; 11.7%, p=0.039). We also found that a significantly higher BMI is linked with hypertension for groups: age 13: BMI (29.2; 33.9, p=0.047), age 14: BMI (24.1; 35.6, p=0.00007) and age 18: BMI (31.8; 45.6, p=0.045). Within the different age groups there were differences between normotensive and hypertensive patients in BMI and HbA1C measurements, but they were not statistically significant and we assume that an increased sample size would be needed to confirm the data. We are currently working on identifying other risk factors including sex, race, height, urine creatinine, urine microalbumin, serum creatinine, lipid profile and thyroid function, that might be responsible for hypertension in pediatric patients with type 2 diabetes.

Acute lung injury (ALI) is characterized by increased endothelial hyperpermeability. Protein nitration is involved in the endothelial barrier dysfunction in LPS-exposed mice. However, the nitrated proteins involved in this process have not been identified. The activation of the small GTPase RhoA is a critical event in the barrier disruption associated with LPS. Thus, in this study we evaluated the possible role of RhoA nitration in this process. Mass spectroscopy identified a single nitration site, located at Tyr(34) in RhoA. Tyr(34) is located within the switch I region adjacent to the nucleotide-binding site. Utilizing this structure, we developed a peptide designated NipR1 (nitration inhibitory peptide for RhoA 1) to shield Tyr(34) against nitration. TAT-fused NipR1 attenuated RhoA nitration and barrier disruption in LPS-challenged human lung microvascular endothelial cells. Further, treatment of mice with NipR1 attenuated vessel leakage and inflammatory cell infiltration and preserved lung function in a mouse model of ALI. Molecular dynamics simulations suggested that the mechanism by which Tyr(34) nitration stimulates RhoA activity was through a decrease in GDP binding to the protein caused by a conformational change within a region of Switch I, mimicking the conformational shift observed when RhoA is bound to a guanine nucleotide exchange factor. Stopped flow kinetic analysis was used to confirm this prediction. Thus, we have identified a new mechanism of nitration-mediated RhoA activation involved in LPS-mediated endothelial barrier dysfunction and show the potential utility of "shielding" peptides to prevent RhoA nitration in the management of ALI.

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Non-obese diabetic (NOD) mice are well-established models of independently developing spontaneous autoimmune diseases, Sjogren’s syndrome (SS) and type 1 diabetes (T1D). The key determining factor for T1D is the strong association with particular MHCII molecule and recognition by diabetogenic T cell receptor (TCR) of an insulin peptide presented in the context of I-Ag7 molecule. For SS the association with MHCII polymorphism is weaker and TCR diversity involved in the onset of the autoimmune phase of SS remains poorly understood. To compare the impact of TCR diversity reduction on the development of both diseases we generated two lines of TCR transgenic NOD mice. One line expresses transgenic TCRβ chain originated from a pathogenically irrelevant TCR, and the second line additionally expresses transgenic TCRαmini locus. Analysis of TCR sequences on NOD background reveals lower TCR diversity on Treg cells not only in the thymus, but also in the periphery. This reduction in diversity does not affect conventional CD4+ T cells, as compared to the TCRmini repertoire on B6 background. Interestingly, neither transgenic TCRβ nor TCRmini mice develop diabetes, which we show is due to lack of insulin B:9–23 specific T cells in the periphery. Conversely SS develops in both lines, with full glandular infiltration, production of autoantibodies and hyposalivation. It shows that SS development is not as sensitive to limited availability of TCR specificities as T1D, which suggests wider range of possible TCR/peptide/MHC interactions driving autoimmunity in SS.

The hypoxia-inducible factor (HIF-1alpha), a basic helix-loop-helix transcription factor, is known to heterodimerize with ARNT1, a nuclear translocator, to trigger the overexpression in many cells of genes involved in resistance to hypoxia. Although HIF-1alpha and ARNT1 are both expressed in brain, their cellular localization and function therein are unknown. Here, using in situ hybridization and immunocytochemistry, we show that HIF-1alpha is expressed in normoxic cerebral neurons together with not only ARNT1 but also ARNT2, a cerebral translocator homologous to ARNT1 but displaying, unlike ARNT1, a selective neuronal expression. In contrast, other potential partners of the translocators, i.e. the aryl hydrocarbon receptor (AHR) and the single-minded protein 2 (SIM2), are not expressed in the adult brain. We also identify two splice variants of HIF-1alpha in brain, one of which dimerizes with ARNT2 even more avidly than with ARNT1. The resulting heterodimer, in contrast with the HIF-1alpha/ARNT1 complex, does not recognize the HIF-1-binding site of the hypoxia-induced erythropoietin (Epo) gene, suggesting that it controls transcription of a distinct set of genes. We therefore propose that HIF-1alpha and ARNT2 function as preferential dimerization partners in neurons to control specific responses, some of which may not be triggered by hypoxia. In support of this proposal, in nonhypoxic PC12 cells constitutively coexpressing HIF-1alpha, ARNT1 and ARNT2, downregulation of either HIF-1alpha or ARNT2, obtained with selective antisense nucleotides, resulted in inhibition of [3H]thymidine incorporation.

The processes responsible for the limited ability to divide and long survival of neurons are not well understood but may involve aryl hydrocarbon receptor nuclear translocator 2 (ARNT2), a recently identified protein, apparently belonging to the basic helix-loop-helix superfamily of transcription factors, which is expressed almost exclusively in brain during the whole lifetime. In agreement, we show, in the rat, that ARNT2 immunoreactivity could be observed only within nuclei of brain neurons and of dividing and neuronal PC12 cells, a localization consistent with a role in transcription regulation. Cell death elicited either by focal ischaemia in brain or oxidative stress in PC12 cells was largely preceded by an almost complete suppression of ARNT2 expression. In contrast, when PC12 cell cycle progression was impaired, ARNT2 expression was enhanced. Finally, the downregulation of ARNT2 levels induced by antisense oligonucleotides prevented PC12 cell proliferation and induced apoptosis. These observations support the hypothesis that ARNT2 is a neuronal transcription factor, regulating cell cycle progression and preventing cell death, whose sustained expression might ensure brain neuron survival.