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Resume Objectifs La population geriatrique, souvent polymedicamentee, est exposee aux risques d’evenements indesirables medicamenteux. La conciliation des traitements medicamenteux (CTM), processus interactif et pluriprofessionnel, est une aide a la continuite des soins. L’objectif de cette etude etait d’analyser et de definir des criteres de priorisation pertinents pour la CTM en geriatrie afin d’eviter le maximum d’erreurs medicamenteuses. Methodes Un audit clinique de la CTM realisee aux points de transition que sont l’admission et la sortie du patient a ete realise de facon prospective pendant 10 mois. Les patients ont ete selectionnes en fonction d’une procedure de priorisation deja etablie dans notre structure qui etait la presence d’au moins un des trois criteres suivants : provenance d’un service hospitalier, insuffisance renale severe et prescription de medicaments a risque. Resultats La cohorte de patients concilies a l’entree comprenait 136 patients. 63 divergences non intentionnelles (DNI) ont ete recensees dont la majeure partie (76,2 %) impliquait des omissions de medicament. Trois criteres ont ete identifies comme facteurs predictifs independants du risque de DNI sur l’ordonnance d’entree par rapport au bilan medicamenteux optimise : antecedents rhumatologiques, provenance d’un service hospitalier et hyponatremie. L’hyponatremie s’est revelee etre le critere qui representait le plus de risque de presenter une DNI sur l’ordonnance du patient, notamment a risque d’omission de traitement a l’entree. Conclusion Cette etude permettra d’ameliorer les criteres de priorisation de la procedure de l’etablissement de sante ainsi que de mettre en place la CTM en hospitalisation de jour geriatrique afin de renforcer le lien ville-hopital.

International audience; Purpose1) To test the hypothesis of the existence of a perinatal vitamin A (VA) programming of VA metabolism and to better understand the intestinal regulation of VA metabolism.MethodsOffspring from rats reared on a control (C) or a VA-deficient (D) diet from 6 weeks before mating until offspring weaning, i.e., 7 weeks after mating, were themselves reared on a C or D diet for 19 weeks, resulting in the following groups: C–C (parents fed C—offspring fed C), D-C, C-D and D-D. VA concentrations were measured in plasma and liver. β-Carotene bioavailability and its intestinal conversion rate to VA, as well as vitamin D and E bioavailability, were assessed after gavages with these vitamins. Expression of genes involved in VA metabolism and transport was measured in intestine and liver.ResultsC–D and D–D had no detectable retinyl esters in their liver. Retinolemia, hepatic retinol concentrations and postprandial plasma retinol response to β-carotene gavage were higher in D–C than in C–C. Intestinal expression of Isx was abolished in C–D and D–D and this was concomitant with a higher expression of Bco1, Scarb1, Cd36 and Lrat in males receiving a D diet as compared to those receiving a C diet. β-Carotene, vitamin D and E bio-availabilities were lower in offspring receiving a D diet as compared to those receiving a C diet.ConclusionA VA-deficient diet during the perinatal period modifies the metabolism of this vitamin in the offspring. Isx-mediated regulation of Bco1 and Scarb1 expression exists only in males severely deficient in this vitamin. Severe VA deficiency impairs β-carotene and vitamin D and E bioavailability.

We compared the efficacy of tailored non pharmacological therapies (NPT) on specific nocturnal behavioral and psychological symptoms of dementia (BPSD). This retrospective 1-year study included 84 older dependent patients institutionalized in 7 long-term care home. Dedicated assistants, who were taught by experts how to use NPT, were asked to record the occurrence of each BPSD episode, to choose a given NPT on the basis of their knowledge of the patient and the type of BPSD and to estimate its efficacy. Wandering was the most prevalent BPSD followed by agitation/aggression and screaming. The most used therapy was cognitive stimulation, followed by multisensory stimulation, reminiscence and Montessori-based. Regarding wandering, multisensory stimulation was found to be the most efficacious NPT significantly different from Montessori-based, cognitive stimulation or reminiscence. With regards to agitation/aggression or screaming, Montessori-based was found to be the most efficacious NPT significantly different from multisensory stimulation, reminiscence and cognitive stimulation.

Modifying the galenic of dry oral forms (DOF) to be administered to patients with swallowing or behavioral disorders is frequent in long term care homes. The objective was to investigate the practice of modifying DOF galenic by nurses in home settings (NHS). A 14-question electronic survey was distributed to 1977 NHS. Almost 3/4 of respondents reported crushing tablets or opening capsules, with 37% on a daily basis. Approximately 22% did not inquire about the feasibility of modifying galenic DOF beforehand. Nearly 75% of NHS were aware about the risk of ineffectiveness after crushing tablets or opening capsules. However, only 14% mentioned the risk of overdosing associated with this practice. More than 60% of NHS never wore protective equipment when changing dosage form. Changing the dosage form of DOF is a widespread practice in home settings. Our present work urges the need to improve collaboration between prescribing physicians, nurses and pharmacists.

Increased platelet activity occurs in type 2 diabetes mellitus (T2DM) and such platelet dysregulation likely originates from altered megakaryopoiesis. We initiated identification of dysregulated pathways in megakaryocytes in the setting of T2DM. We evaluated through transcriptomic analysis, differential gene expressions in megakaryocytes from leptin receptor-deficient mice (db/db), exhibiting features of human T2DM, and control mice (db/+). Functional gene analysis revealed an upregulation of transcripts related to calcium signaling, coagulation cascade and platelet receptors in diabetic mouse megakaryocytes. We also evidenced an upregulation (7- to 9.7-fold) of genes encoding stefin A (StfA), the human ortholog of Cystatin A (CSTA), inhibitor of cathepsin B, H and L. StfA/CSTA was present in megakaryocytes and platelets and its expression increased during obesity and diabetes in rats and humans. StfA/CSTA was primarily localized at platelet membranes and granules and was released upon agonist stimulation and clot formation through a metalloprotease-dependent mechanism. StfA/CSTA did not affect platelet aggregation, but reduced platelet accumulation on immobilized collagen from flowing whole blood (1200 s−1). In-vivo, upon laser-induced vascular injury, platelet recruitment and thrombus formation were markedly reduced in StfA1-overexpressing mice without affecting bleeding time. The presence of CA-074Me, a cathepsin B specific inhibitor significantly reduced thrombus formation in-vitro and in-vivo in human and mouse, respectively. Our study identifies StfA/CSTA as a key contributor of platelet-dependent thrombus formation in both rodents and humans.

Purpose Older patients with colon cancer (CC) are vulnerable to chemotherapy toxicity and death. Establishing simple scores specific for patients with CC to predict severe chemotoxicity or early death is needed to select the best treatment strategy. Subjects, Materials, and Methods This prospective multicenter study included patients aged ≥70 years with CC receiving adjuvant or first-line metastatic chemotherapy. Frailty markers (nutrition, physical activity, energy, mobility, strength), comprehensive geriatric assessment (functional status, comorbidities, falls, nutrition, cognition, and depression), and usual laboratory parameters were collected. Logistic or Cox regression was used to examine at 500 days the association between frailty markers, comprehensive geriatric assessment, laboratory parameters, and grade 3–4 toxicity or death. Results A total of 97 patients (median age, 79.0 years) received adjuvant (37.1%) or metastatic (62.9%) chemotherapy. During the first 500 days, grade 3–4 toxicity occurred in 49.5%, and 30% died. The predictive model for grade 3–4 toxicity combined (polychemotherapy × 3) + (hypoalbuminemia 11 mg/L + Eastern Cooperative Oncology Group performance status (ECOG-PS), cutoff score >3. The predictive model for death combined (metastasis × 5) + (age × 2) + alkaline phosphatase >100 IU/mL + sex (female) + abnormal grip strength + ECOG-PS, cutoff score >6. For chemotoxicity prediction, sensitivity was 81.6% and specificity 71.4%. For death prediction, sensitivity was 89.7% and specificity was 83.6%. Conclusion These simple and efficient “ColonPrediscores” will help to better identify older patients with CC with increased risk of chemotherapy-related toxicity and/or death.

Chronic heart failure (CHF) is a major public health matter. Mainly affecting the elderly, it is responsible for a high rate of hospitalization due to the frequency of acute heart failure (ADHF). This represents a disabling pathology for the patient and very costly for the health care system. Our study is designed to assess a connected and portable bioelectrical impedance analysis (BIA) that could reduce these hospitalizations by preventing early ADHF. METHODS This prospective study included patients hospitalized in cardiology for ADHF. Patients achieved 3 self-measurements using the BIA during their hospitalization and answered a questionnaire evaluating the acceptability of this self-measurement. The results of these measures were compared with the clinical, biological and echocardiographic criteria of patients at the same time. RESULTS Twenty-three patients were included, the self-measurement during the overall duration of the hospitalization was conducted autonomously by more than 80% of the patients. The acceptability (90%) for the use of the portable BIA was excellent. Some correlations were statistically significant, such as the total water difference to the weight difference (p=0.001). There were common trends between the variation of impedance analysis measures and other evaluation criteria. CONCLUSION The feasibility and acceptability of a self-measurement of bioelectrical impedance analysis by the patient in AHF opens up major prospects in the management of monitoring patients in CHF. The interest of this tool is the prevention of ADHF leading to hospitalization or re-hospitalizations now requires to be presented by new studies.

International audience; Scope The effect of vitamin A deficiency on vitamin A and lipid postprandial metabolism in young rats is addressed, considering the effect of sex. Methods and Results Sprague-Dawley rats are fed either 400 UI.kg(-1) vitamin A diet (vitamin A-deficient (VAD) diet) or 2300 UI.kg(-1) vitamin A (control diet), before being mated. Mothers receive the same VAD or control diet during gestation and lactation. Offspring receive the same diet than mothers until 8 weeks of age. VAD diet-fed female and male offspring display a severe vitamin A deficiency with no body weight or glucose tolerance defects. Fasting plasma triglyceride concentrations are decreased in VAD diet-fed animals compared to controls (p < 0.05). Retinyl ester postprandial responses after vitamin A gavage, expressed as area under the curves, are not different in VAD diet-fed and control animals, although retinyl ester postprandial peak is significantly delayed (p < 0.05) in VAD diet-fed rats. Lipids also accumulate in the distal part of the intestine after gavage and [1-C-13]-oleate postprandial response is decreased in VAD diet-fed males. Conclusion Vitamin A deficiency modulates both vitamin A absorption rate and lipid postprandial metabolism, which can partly explain the altered fasting lipid status observed in VAD diet-fed offspring.

Purpose: Since intrathoracic goiters (IG), either cervico-mediastinal goiters (CMGs) or mediastinal nodules (MNs), can lead to sternotomies and/or evitable reoperations, their detection is mandatory before thyroid surgery. A systematic screening by CT scan or MRI is not conceivable because of their expensiveness. We tested if conventional chest radiography (CCR) could remain a good screening tool for IG before thyroid surgery. Methods: In this retrospective study (2554 patients), CCR usefulness was evaluated in relation with patients’ complaints, clinical examination, neck US, and anatomical and surgical findings. Results: CMGs (n = 67) and MNs (n = 42) were symptomatic in 10 and 5 patients, respectively. Clinical examination or neck US suspected their existence in 25 and 13 and 45 and 17 patients, respectively. Among the 50 IG detected by CCR (42 CMGs and 8 MNs), 4 CMGs and 2 MNs were missed by clinical examination or neck US. CCR failed to detect IG in 59 patients (54%): 25 CMGs (37%) and 34 MNs (80%). Twenty-eight IG (9 CMGs and 19 MNs) were discovered during surgery. CCR resulted in false positive in 88 out of 2445 patients (3.5%). CCR potentially avoided reoperation in two patients (a maximum saving of 6160 €, whereas the total cost of CCR was 54,895 €). Conclusions: CCR should not be used routinely for the preoperative detection of IG. Surgeons should preferably use clinical examination or neck US and directly perform CT scan when a mediastinal extension is suspected.

Background Flow cytometry essentially focuses on surface-expressed proteins, with few protocols being devoted to intracellular components. We evaluated a two-step procedure using new formaldehyde-free permeabilization and staining reagents that allow the staining of platelets and red blood cells (RBCs) from whole blood. Methods Citrated blood was treated with the new staining protocol (NSP) or control reagent (phosphate-buffered solution bovine serum albumin) and stained with antibodies against surface or intracellular markers. The effects of the NSP on cell integrity, morphology, and content were evaluated. Results The NSP slightly reduced the cell count (~20%) and changed the RBC morphology with a 42% mean diameter reduction. Conversely, the NSP did not affect platelet discoid morphology and led to a minor size decrease (11%). These morphological changes neither impelled a gating strategy modification nor interfered with the discrimination among populations based on surface markers. The NSP provided intracellular access to all the tested antigens: CD62P, FXIII, and CD63 in platelets and glycated and fetal hemoglobin (HbA1c and HbF) and nucleic acid in RBCs. The NSP gave excellent intra-assay precision with minimal impact on cell morphology and fluorescence labelling over time (up to 24 h). Conclusions With the ability to detect surface and intracellular antigens through a rapid preparation protocol without washing steps or toxic formaldehyde treatment, this NSP designed for research offers a marked improvement in the analysis of platelets and RBCs isolated directly from whole blood. Consequently, the NSP opens new avenues to investigate platelet degranulation and erythrocyte subpopulations. © 2019 International Clinical Cytometry Society.

Resume Introduction La non-observance du traitement substitutif de l’hypothyroidie est la cause la plus frequente de la discordance entre une TSH elevee malgre une forte posologie de L-thyroxine. Observation La TSH variait de 100 a 400 mUI/L chez un patient de 36 ans traite par L-thyroxine et L-triiodothyronine (respectivement 300 et 75 μg/jour) pour une hypothyroidie postchirurgicale. L’absorption de L-thyroxine etait normale apres charge orale. La TSH etait par la suite normalisee par une prise orale hebdomadaire de 1000 μg de L-thyroxine. Conclusion Le test de charge orale en L-thyroxine est indispensable au diagnostic apres avoir elimine les autres causes classiques : interactions medicamenteuses, malabsorption digestive et erreur de dosage. La prise en charge est a discuter au cas par cas : prise quotidienne du traitement sous surveillance, prise hebdomadaire d’une forte dose (1000 μg) de L-thyroxine ou injection intramusculaire de la meme posologie (sous reserve de l’accord de l’assurance maladie).

Introduction Les mastocytoses font partie des maladies rares potentiellement associees a de nombreux symptomes systemiques. Ces symptomes sont responsables d’un handicap parfois marque avec des consequences socioprofessionnelles pouvant etre importantes [1] . L’hypnose medicale est actuellement utilisee avec succes dans de nombreuses specialites medicales telles que la chirurgie, la psychiatrie, l’oncologie, la rhumatologie, etc. L’objectif principal de cette etude etait de determiner l’impact de seances d’hypnose medicale sur les symptomes et la qualite de vie de patients atteints de mastocytoses avec symptomes invalidants. Patients et methodes Cette etude prospective de mai 2016 a mars 2019 a inclus des patients atteints de mastocytose agressive (MA), mastocytose systemique indolente (MS) ou mastocytose cutanee (MC), presentant des symptomes invalidants malgre un traitement optimise (traitements symptomatiques ou traitements cytoreducteurs ou therapies ciblees pour les formes les plus graves) et stable. Deux seances d’hypnose medicale ont ete programmees avec un intervalle de 8 a 15 jours. Les donnees collectees comprenaient : âge, sexe, type de mastocytose, symptomes de la maladie. Les scores de qualite de vie (QLQ30), de symptomes intestinaux (score derive du CDAI [2] ), 3 scores bases sur l’echelle visuelle analogique (EVA) (douleur globale, energie, flushs) et le score AFFIRM [1] ont ete recueillis 8 jours avant la 1re seance, 8 jours avant la 2e, 8 jours apres la 2e seance et un mois apres la 2e seance. Le Patient Global Impression of Change (PGIC) score [3] a ete recueilli en juillet 2019. Resultats Vingt patients (moyenne d’âge 53,3 ans, 75 % de femmes, 15 MS, 1 MA et 4 MC, duree moyenne de la maladie 14,9 annees) ont ete inclus. Tous les patients presentaient une asthenie et des douleurs musculosquelettiques. Les autres symptomes les plus frequents etaient les douleurs abdominales (18/20), le prurit (17/20), les cephalees (17/20) et les flushs (17/20). Les 3 items suivants du CDAI : nombre de jours avec douleurs abdominales, intensite des douleurs abdominales, bien etre general de meme que la fatigue etaient significativement ameliores apres la 1re et apres la 2e seance (respectivement p = 0,0352 et p = 0,0052, p = 0,0492 et p = 0,0219, p = 0,0363 et p = 0,0482, p = 0,0340 et p = 0,0393). Un mois apres la fin des 2 seances, l’intensite des douleurs abdominales et la fatigue restaient ameliores alors que le nombre de jours avec douleurs abdominales et le bien etre general etaient revenus a l’etat basal. Le score AFFIRM etait significativement ameliore durant toute l’etude (p = 0,0075). Il etait significativement diminue apres la 2e seance (p = 0,0267) et restait ameliore un moins apres (p = 0,0214). Le PGIC montrait que l’amelioration etait durable plus de 4 mois apres la fin de l’etude chez 8/20 (score entre 5 et 7). Conclusion Chez les patients atteints de mastocytose, l’hypnose permet une amelioration de symptomes invalidants et l’impact de 2 seances d’hypnose persiste a un mois. Certains patients gardent une amelioration a long terme. L’hypnose semble etre une therapie complementaire utile dans cette maladie orpheline dont les symptomes sont tres invalidants et parfois difficiles a controler par les medicaments actuels.

Objectives To determine whether environmental rearrangements of the long-term care nursing home can affect disruptive behavioral and psychological symptoms of dementia (BPSD) in residents with dementia. Design Prospective 6-month study. Setting The study was conducted before (phase 1) and after (phase 2) environmental rearrangements [skylike ceiling tiles in part of the shared premises, progressive decrease of the illuminance at night together with soothing streaming music, reinforcement of the illuminance during the day, walls painted in light beige, oversized clocks in corridors, and night team clothes color (dark blue) different from that of the day team (sky blue)]. Participants All of the patients (n = 19) of the protected unit were included in the study. They were aged 65 years or older and had an estimated life expectancy above 3 months. Measures Number and duration of disruptive BPSD were systematically collected and analyzed over 24 hours or during late hours (6:00-12:00 pm ) during each 3-month period. Results There was no significant change in the patients' dependency, risk of fall, cognitive or depression indexes, or treatment between phase 1 and 2. Agitation/aggression and screaming were observed mainly outside the late hours as opposed to wandering episodes that were noticed essentially within the late hours. The number of patients showing wandering was significantly lower over 24 hours during phase 2. The number of agitation/physical aggression, wandering, and screaming and the mean duration of wandering episodes were significantly (P = .039, .002, .025, and .026 respectively) decreased over 24 hours following environmental rearrangements. Similarly, a significant reduction in the number and mean duration of wandering was noticed during the late hours (P = .031 and .007, respectively). Conclusions Our study demonstrates that BPSD prevalence can be reduced following plain environmental rearrangements aimed at improving spatial and temporal orientation.

International audience

International audience; Objective: To describe glycemic control in nursing home residents with diabetes and to evaluate the relevance of HbA1c in the detection of hypoglycemia risk.Design and methods: Diabetes treatment, geriatric assessment, blood capillary glucose (n= 24,682), and HbA1c were collected from medical charts of 236 southern France nursing home residents during a 4-month period. Glycemic control was divided into four categories: tight, fair, and moderate or severe chronic hyperglycemia using the High Blood Glucose Index or the analysis of blood glucose frequency distribution. Hypoglycemia episodes were identified by medical or biological records.Results: Glucose control was tight in 59.3 % and fair in 19.1 % of the residents. Chronic exposure to hyperglycemia was observed in 21.6 % of the residents (severe in 9.7 % and moderate in 11.9 %). Hypoglycemia was noticed in 42/236 (17.8%), in all categories of glycemic control. Relative hypoglycemia risk was significantly (P = 0.0095) higher in residents with moderate chronic hyperglycemia compared with those with tight control. The majority of residents with hypoglycemia (39/42) or chronic hyperglycemia (45/51) were insulin-treated. The relative risk of hypoglycemia was not significantly associated with HbA1c values.Conclusion: Hypoglycemia risk in nursing home residents is observed in all categories of glycemic control. In tight control, the potency of antidiabetic treatment should be reduced. In chronic hyperglycemia, diet and treatment should be reevaluated in order to reduce glucose variability. HbA1c is not sufficient for hypoglycemia risk detection; capillary blood glucose monitoring is warranted for nursing homes residents with diabetes.

Recently, two clinical trials about a "sensor-based flash glucose monitoring system" and its efficacy in reducing time in hypoglycemia were published. Interestingly, patients spent more time at low glucose concentrations in these studies than in other studies related to the efficacy of real-time continuous glucose monitoring (rtCGM). Although it is possible that the study populations differed from those in other studies, another potential explanation is that the CGM system used in these two studies had a negative glucose measurement bias. Such a negative bias was reported in recent literature, suggesting that the CGM system may inaccurately indicate hypoglycemia. Reporting blood glucose monitoring data would help to interpret the CGM data at least in the context of time spent in various glucose ranges as a parameter with which quality of diabetes therapy is measured.

International audience; no abstract

International audience; no abstract

International audience; Alterations in the nutritional perinatal environment, such as intrauterine growth retardation with subsequent postnatal catch-up growth, program cardiovascular disease in adulthood, possibly through alterations in matrix metalloproteinase (MMP)-2 and -9. However, experimental evidences demonstrating that changes in the nutritional perinatal environment can program MMP-2 and -9 with subsequent alterations of vessel wall are lacking.; The current study evaluated whether immediate postnatal overfeeding is able to alter vascular morphological indexes and circulating and/or vascular MMP2-2 and -9 status.; Aortic morphology (wall thickness and percentage of incomplete elastin lamellae) and circulating and aortic MMP-2 and -9 activity (measured by gelatin zymography) and aortic MMP-2 and -9 mRNA (measured by reverse transcription polymerase chain reaction (RT-PCR)) were studied in adult male rats overfed (OF) or normofed (NF) during the immediate postnatal period.; Postnatal overfeeding induced early onset obesity. Adult OF rats presented with increased blood pressure and circulating MMP-2 and -9 activity. In the thoracic aorta, postnatal overfeeding increased wall thickness and decreased elastin integrity (as demonstrated by an increased percentage of incomplete elastin lamellae). OF rats showed enhanced aortic MMP-2 activity and MMP-9 mRNA levels. Circulating and aortic MMP-2 activity correlated positively with the percentage of incomplete elastin lamellae and aortic wall thickness, respectively.; Our data demonstrate for the first time that immediate postnatal nutritional programming induces increases in circulating and aortic MMP-2 activity with parallel aortic wall alterations, such as decreased elastin integrity and enhanced thickening, showing that this experimental model is suitable for the study of perinatal nutritional programming of vascular functions.

Objective— High endogenous thrombin potential (ETP) is associated with venous and arterial thrombosis. Better knowledge of environmental influences on ETP may help to prevent thrombosis. Methods and Results— Weaning rats exhibited high ETP values that decreased in low-fat diet and remained elevated on high-fat diet. In adult rats, high-fat diet–induced ETP increase was independent of coagulation factors, obesity, and insulin resistance and negatively associated with polyunsaturated fatty acid levels. Switching from high-fat diet to low-fat diet reversed the procoagulant phenotype with a slower kinetic than the normalization of hyperinsulinemia. In humans, ETP was independent of body weight whereas it was negatively associated with nutritional markers such as the percentage of energy provided by proteins, the protein:fat ratio, circulating phenolic compounds, and omega-3 polyunsaturated fatty acid. A recommended 3-month healthy diet with reduced energy density, including lipids, decreased ETP (−21%; P Conclusion— Diet plays a pivotal role in regulating ETP, independently of obesity and insulin resistance. Global nutritional recommendations could be useful in primary prevention of venous thrombosis.

10041Background: Older patients with colon cancer are more vulnerable to chemotherapy toxicity and early death. Establishing simple scores specific for colon cancer (cc) patients able to predict se...

Epicardial fat is a relatively neglected component of the heart and could be an important risk factor of cardiac disease. The objective of our study was to assess the relationship between epicardial adipose tissue (EAT) extent, fat distribution, and coronaropathy in a group of adult victims of accidental or suspicious sudden death. In 56 cadavers, we performed 34 measurements of EAT from five computerized photographs of the heart (anterior and posterior faces, and three ventricle transversal slices) and analyzed their relationship with anthropometric markers of adiposity (BMI, waist and leg circumference, thickness of abdominal and thigh subcutaneous adipose tissue (SAT)), with the presence and staging of coronary artery disease (CAD), and with markers of myocardial hypertrophy. Simple linear regressions showed that EAT measurements are highly intercorrelated (r from 0.4 to 0.6, P < 0.001), and correlate with age, waist circumference, and heart weight, and to a lesser extent, with BMI, abdominal SAT thickness, and leg SAT thickness. Multiple regression showed that age, waist circumference, and heart weight significantly and independently correlate with EAT (P < 0.0001). No other anthropometric measurement was found independently correlated with EAT. The EAT/myocardium ratios correlated positively with age and waist circumference. Anterior and posterior areas of EAT were found significantly increased in patients with CAD and correlated positively with CAD staging (P = 0.0034, r = 0.38). Anterior EAT surface was found positively associated with CAD (P = 0.01), independently of age and other adiposity measurements. Prospective studies are needed to assess the risk of occurrence/progression of CAD that relate to EAT excess.

Recently, we characterized tumor suppressor candidate 5 (Tusc5) as an adipocyte-neuron PPARgamma target gene. Our objective herein was to identify additional genes that display distinctly high expression in fat and neurons, because such a pattern could signal previously uncharacterized functional pathways shared in these disparate tissues. gamma-Synuclein, a marker of peripheral and select central nervous system neurons, was strongly expressed in white adipose tissue (WAT) and peripheral nervous system ganglia using bioinformatics and quantitative PCR approaches. Gamma-synuclein expression was determined during adipogenesis and in subcutaneous (SC) and visceral adipose tissue (VAT) from obese and nonobese humans. Gamma-synuclein mRNA increased from trace levels in preadipocytes to high levels in mature 3T3-L1 adipocytes and decreased approximately 50% following treatment with the PPARgamma agonist GW1929 (P < 0.01). Because gamma-synuclein limits growth arrest and is implicated in cancer progression in nonadipocytes, we suspected that expression would be increased in situations where WAT plasticity/adipocyte turnover are engaged. Consistent with this postulate, human WAT gamma-synuclein mRNA levels consistently increased in obesity and were higher in SC than in VAT; i.e. they increased approximately 1.7-fold in obese Pima Indian adipocytes (P = 0.003) and approximately 2-fold in SC and VAT of other obese cohorts relative to nonobese subjects. Expression correlated with leptin transcript levels in human SC and VAT (r = 0.887; P < 0.0001; n = 44). Gamma-synuclein protein was observed in rodent and human WAT but not in negative control liver. These results are consistent with the hypothesis that gamma-synuclein plays an important role in adipocyte physiology.

Epicardial white adipose tissue (eWAT) is in close contact with coronary vessels and therefore could alter coronary homeostasis. Adrenomedullin (AM) is a potent vasodilatator and antioxidative peptide which has been shown to play a cytoprotective role in experimental models of acute myocardial infarction. We studied, using immunohistochemistry and qRT-PCR, the expression of AM and its receptors calcitonin receptor-like receptor (CRLR), and receptor activity-modifying protein (RAMP)2 and -3 in paired biopsies of subcutaneous WAT (sWAT) and eWAT obtained from patients with coronary artery disease (CAD) or without CAD (NCAD). In eWAT obtained from NCAD or CAD patients, immunoreactivity for AM, CRLR, and RAMP2 and -3 was detected in blood vessel walls and isolated stromal cells close to adipocytes. Some of the AM positive stromal cells colocalized CD68 immunoreactivity. eWAT from CAD patients showed increased AM immunoreactivity and AM gene expression. CRLR mRNA levels were comparable in sWAT of both groups and decreased by 40–50% in eWAT, irrespectively of the coronary status. RAMP2 mRNA concentrations did not change while RAMP3 mRNA levels increased in sWAT from CAD patients. There was a positive linear relationship between eWAT 11β-hydroxysteroid dehydrogenase type 1 mRNA (11β-HSD-1, the enzyme that converts inactive to active glucocorticoids) and AM mRNA. In conclusion, we demonstrate that AM and its receptors are expressed in eWAT. Our data suggest that eWAT AM, which could originate from macrophages, is related to 11β-HSD-1 expression. AM synthesis, which is increased in eWAT during chronic CAD in humans, can play a cardioprotective role.

It has been hypothesized that sleep apnea syndrome (SAS) increases hypothalamic-pituitary-adrenal axis activity and, through increased cortisol levels, participates in the pathophysiology of metabolic and cardiovascular complications. We compared the circadian profiles of cortisol in obese men with [obSAS+; apnea-hypopnea index (AHI) ≥ 20/h] and without SAS (obSAS−; AHI ≤ 5/h). 1) Salivary cortisol (5 samples: before/30 min after dinner, 2100, upon/30 min after awakening) was measured in 15 obSAS+, 19 obSAS−, and 19 normal-weight controls (NWC). 2) Plasma cortisol (every 30 min for 24 h under highly controlled conditions and portable EEG device) was measured in 9 obSAS+, 8 obSAS−, and 10 NWC men. Visceral adipose tissue surface was measured by CT scan. In both studies, obSAS+ and obSAS− men were comparable for age, BMI, waist circumference, and waist-to-hip ratio. First, no difference was found, using ANOVA for repeated measures, between obSAS+ and obSAS− subjects for any salivary cortisol measurement. No correlation was found between salivary cortisol and AHI or nocturnal SaO2. Similarly, obSAS+ and obSAS− men showed no difference in plasma cortisol rhythmicity: 24-h minimum, maximum, and mean, ANOVA for repeated measures, mathematical modeling of cortisol rhythm (COSINOR), and morning secretory peak. Conversely, ANOVA for repeated measures showed decreased cortisol levels in obese vs. NWC men during both the trough (2200–0130) and the peak (0600–0900) independently of SAS status. We show that SAS per se is not associated with any change of the level or of the features of salivary and plasma cortisol rhythmicity and confirm that men with visceral obesity display lower plasma cortisol levels than NWC men.

International audience; Immediate postnatal overfeeding in rats, obtained by reducing the litter size, results in early-onset obesity. Such experimental paradigm programs overweight, insulin resistance, dyslipidemia, increased adipose glucocorticoid metabolism [up-regulation of glucocorticoid receptor (GR) and 11 beta-hydroxysteroid dehydrogenase type 1 (11 beta-HSD1)], and overexpression of proinflammatory cytokines in mesenteric adipose tissue (MAT) in adulthood. We studied the effects of pioglitazone, a PPAR gamma agonist, treatment on the above-mentioned overfeeding-induced alterations. Nine-month-old rats normofed or overfed during the immediate postnatal period were given pioglitazone (3 mg/kg/day) for 6 weeks. Pioglitazone stimulated weight gain and induced a redistribution of adipose tissue toward epididymal location with enhanced plasma adiponectin. Treatment normalized postnatal overfeeding-induced metabolic alterations (increased fasting insulinemia and free fatty acids) and mesenteric overexpression of GR, 11 beta-HSD11, CD 68, and proinflammatory cytokines mRNAs, including plasminogen-activator inhibitor type 1. MesentericGRmRNAlevels correlated positively with mesenteric proinflammatory cytokinesmRNAconcentrations. In vitro incubation of MAT obtained from overfed rats demonstrated that pioglitazone induced a down-regulation of GR gene expression and normalized glucocorticoid-induced stimulation of 11 beta-HSD1 and plasminogen-activator inhibitor type 1 mRNAs. Our data show for the first time that the metabolic, endocrine, and inflammatory alterations induced by early-onset postnatal obesity can be reversed by pioglitazone at the adulthood. They demonstrate that pioglitazone, in addition to its well-established effect on adipose tissue redistribution and adiponectin secretion, reverses programing-induced adipose GR, 11 beta-HSD1, and proinflammatory cytokines overexpression, possibly through a GR-dependent mechanism.

Glucocorticoids hypersensitivity may be involved in the development of abdominal obesity and insulin resistance. Eight normal weight and eight obese women received on two occasions a 3-h intravenous infusion of saline or hydrocortisone (HC) (1.5 μg·kg−1·min−1). Plasma cortisol, insulin, and glucose levels were measured every 30 min from time−30(min) (time−30) to time240. Free fatty acids, adiponectin, and plasminogen activator inhibitor-1 (PAI-1) levels were measured at time−30, time180, and time240. At time240, subjects underwent an insulin tolerance test to obtain an index of insulin sensitivity (KITT). Mean30–240cortisol level was similar in control and obese women after saline (74 ± 16 vs. 75 ± 20 μg/l) and HC (235 ± 17 vs. 245 ± 47 μg/l). The effect of HC on mean180–240insulin, mean180–240insulin resistance obtained by homeostasis model assessment (HOMA-IR), and KITTwas significant in obese (11.4 ± 2.0 vs. 8.2 ± 1.3 mU/l, P < 0.05; 2.37 ± 0.5 vs. 1.64 ± 0.3, P < 0.05; 2.81 ± 0.9 vs. 3.32 ± 1.02%/min, P < 0.05) but not in control women (3.9 ± 0.6 vs. 2.8 ± 0.5 mU/l; 0.78 ± 0.1 vs. 0.49 ± 0.1; 4.36 ± 1.1 vs. 4.37 ± 1.2%/min). In the whole population, the quantity of visceral fat, estimated by computerized tomography scan, was correlated with the increment of plasma insulin and HOMA-IR during HC infusion [Δmean30–240insulin ( r = 0.61, P < 0.05), Δmean30–240HOMA-IR ( r = 0.66, P < 0.01)]. The increase of PAI-1 between time180and time240after HC was higher in obese women (+25%) than in controls (+12%) ( P < 0.05), whereas no differential effect between groups was observed for free fatty acids or adiponectin. A moderate hypercortisolism, equivalent to that induced by a mild stress, has more pronounced consequences on insulin sensitivity in abdominally obese women than in controls. These deleterious effects are correlated with the amount of visceral fat.

Summary. Background: Plasminogen activator inhibitor type 1 (PAI-1) is the main inhibitor of the fibrinolytic system and contributes to an increased risk of atherothrombosis in insulin-resistant obese patients. In adipose tissue, we have shown that PAI-1 is synthesized mainly in the visceral stromal compartment and is positively regulated by glucocorticoids. We have demonstrated that adipose tissue expression of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD-1), an enzyme that catalyzes the conversion of inactive cortisone to active cortisol, is exaggerated in obese patients. Objectives: We hypothesized that increased action of 11β-HSD-1 in adipose tissue of obese subjects may contribute to PAI-1 overproduction. Patients and methods: Using in situ hybridization, we studied the expression of the mRNAs coding for PAI-1 and 11β-HSD-1 in the stromal compartment of visceral adipose tissue obtained from obese women. The regulation of PAI-1 secretion from in vitro incubated tissue explants was also investigated. Results: Regression analysis showed a significant positive linear relationship between PAI-1 and 11β-HSD-1 mRNAs expression. In vitro incubation of adipose tissue explants demonstrated that cortisone stimulated PAI-1 gene expression and secretion, and that these effects were inhibited by co-incubation with the 11β-HSD inhibitor, glycyrrhetinic acid. Conclusions: Our data demonstrate that 11β-HSD-1-driven cortisone reactivation regulates adipose PAI-1 synthesis and secretion. They suggest that the increased PAI-1 synthesis and secretion observed in obese patients can be also related, at least in part, to an increased local conversion of cortisone to cortisol. Therefore, local cortisol metabolism in adipose tissue may be involved in increasing the risk of cardiovascular disease in obese subjects.

In humans, a hyperactivity of glucocorticoid metabolism was postulated to be involved in the intrauterine programming of the metabolic syndrome in adulthood. We studied in rats the effects of overfeeding, obtained by reducing the size of the litter in the immediate postnatal period, a time crucial for neuroendocrine maturation such as late gestation in humans. Overfeeding induced early-onset obesity and accelerated the maturation of the hypothalamo-pituitary-adrenal (HPA) axis together with an upregulation of adipose tissue glucocorticoid receptor (GR) mRNA. In adulthood, neonatally overfed rats presented with moderate increases in basal and stress-induced corticosterone secretion and striking changes in visceral adipose tissue glucocorticoid signaling, that is, enhanced GR and 11β-hydroxysteroid dehydrogenase type 1 mRNA levels. The above-mentioned alterations in the endocrine status of overfed rats were accompanied by a moderate overweight status and significant metabolic disturbances comparable to those described in the metabolic syndrome. Our data demonstrate for the first time that postnatal overfeeding accelerates the maturation of the HPA axis and leads to permanent upregulation of the HPA axis and increased adipose tissue glucocorticoid sensitivity. Thus, the experimental paradigm of postnatal overfeeding is a powerful tool to understand the pathophysiology of glucocorticoid-induced programming of metabolic axes.

Objective— Because obesity and insulin resistance (IR) are strongly associated with liver steatosis (LS), we investigated the relation between the degree of LS and plasminogen activator inhibitor-1 (PAI-1) in ob/ob mice, in C57/BL6 mice with alcoholic LS, and in severely obese humans. Methods and Results— In both mouse models, plasma PAI-1 levels were associated with PAI-1 expression in the liver and with the degree of LS. Liver PAI-1 antigen was associated with the tumor necrosis factor receptor-II (TNFRII) antigen, whereas association with TNF antigen content was found in ob/ob mice only. No significant correlation between plasma PAI-1 and PAI-1 expression in adipose tissue of ob/ob mice was observed. Furthermore, the relation between plasma PAI-1 levels and body weight was positive in ob/ob mice but negative in C57/BL6 mice (both P Conclusions— Plasma PAI-1 levels are more closely related to fat accumulation and PAI-1 expression in the liver than in adipose tissue. In steatotic liver, PAI-1 antigen content is associated with those of TNF and TNFRII. Therefore, we suggest that TNF pathway dysregulation in LS could be involved in increased plasma PAI-1 in obesity with IR.

Introduction Les effets benefiques de la prise en charge de l’hypercholesterolemie et des statines en prevention secondaire des pathologies cardiovasculaires chez l’adulte d’âge moyen constituent la base de recommandations precises. Il n’en est pas de meme des personnes de plus de 75 ans et dependantes et des residents en EHPAD chez qui la prise en charge doit etre « personnalisee ». Cependant, les pathologies cardiovasculaires y sont la premiere cause de mortalite. L’objectif de cette etude observationnelle est d’evaluer les modalites de prise charge des lipides en prevention cardiovasculaire secondaire chez des residents en EHPAD et de determiner s’il y’a une relation entre ces modalites et les donnees de l’evaluation geriatrique standardisee (EGS). Patients et methodes L’etude a ete realisee chez 961 residents dans 4 EHPAD publiques et 8 EHPAD prives de Marseille et environs. Les donnees suivantes ont ete collectees dans les dossiers medicaux : items de l’EGS, pathologie atheromateuse (coronaropathie, accident vasculaire cerebral, arteriopathie peripherique), diabete, dosage des lipides sanguins au cours de l’annee, traitement par statine, antiagregant, antihypertenseur. Resultats Au total, 410/961 residents (42 %), etaient eligibles pour une prevention cardiovasculaire secondaire : diabete n = 190 (46 %) dont la moitie avait une pathologie atheromateuse ; AVC 33 %, coronaropathie 25 %, arteriopathie peripherique 12 %. Les lipides ont ete doses chez 47 % des residents et un traitement hypolipemiant prescrit a 26 % des residents avec une bonne tolerance. Un tiers des residents sous statine n’ont pas eu de suivi annuel du bilan lipidique. Il n’y avait pas de difference significative dans la prescription de statine selon la pathologie atheromateuse ou la presence de diabete. La prescription de statine etait significativement correlee avec l’avancee en âge et aux donnees de l’EGS (severite de la dependance, denutrition). Conclusion Ces resultats indiquent qu’il n’y a pas d’homogeneite dans la prescription de statine et le suivi des lipides sanguins en prevention secondaire de la pathologie atheromateuse chez les personnes âgees dependantes en EHPAD. Il est necessaire d’evaluer le rapport benefice/risque de la prise en charge des lipides en prevention secondaire dans cette population avec polypathologie, polymedication et dont l’esperance de vie est limitee.

The pituitary-specific POU homeodomain factor Pit-1 likely interacts with other factors for cell-specific expression of prolactin. Here we identify the paired-like homeobox transcription factors Pitx1 and Pitx2 as factors functionally activating the proximal human prolactin promoter (hPRL-164luc). Using in vitro binding assays and a series of site-specific mutations of the proximal hPRL promoter, we mapped the B1 and B2 bicoid sites involved in Pitx-mediated transactivation of the hPRL-164luc construct. In somatolactotroph GH4C1 cells, basal proximal hPRL promoter activity was inhibited by a Pitx2 dominant-negative form in a dose-dependent manner, whereas binding disruptive mutations in the Pitx sites significantly reduced basal activity of the promoter. We also show that synergistic activation of hPRL-164luc by Pitx2 and Pit-1 requires the integrity of the B2 Pitx binding site, and at least one of the P1 and P2 Pit-1 response elements. In addition, mutation in the B2 Pitx site results in attenuation of the promoter's responsiveness to forskolin, thyrotropin-releasing hormone, and epidermal growth factor. Conversely, Pitx1 or Pitx2 overexpression in GH4C1 cells leads to an enhancement of the drugs stimulatory effects. Altogether, these results suggest that full responsiveness to several signaling pathways regulating the hPRL promoter requires the B2 Pitx binding site and that Pitx factors may be part of the proteic complex involved in these regulations. Finally, in situ hybridization analysis showing coexpression of the PRL and Pitx2 genes in rat and human lactotroph cells corroborates the physiological relevance of these results.

The vasopressin V3 receptor (V3) is specifically expressed in pituitary corticotropes and mediates the stimulatory effect of vasopressin on adrenocorticotropic hormone (ACTH) release. The V3 gene is overexpressed in corticotrope pituitary tumours compared to normal pituitaries. We hypothesized that V3 overexpression might induce changes in corticotrope function and alter the regulation of the hypothalamic-pituitary-adrenal axis. Thus, we generated transgenic mice (POMV3) expressing the human V3 receptor in the pituitary under the control of rat pro-opiomelanocortin (POMC) promoter sequences. The transgene was efficiently transcribed and vasopressin binding was increased in both corticotropes and melanotropes. In-vitro ACTH release and inositol phosphate formation were unchanged in POMV3 pituitaries, but the responses to vasopressin were significatively increased. In vivo, basal circulating concentrations of ACTH in POMV3 mice were similar to those of controls but corticosterone concentrations were moderately increased. In addition, the levels of POMC mRNA in the transgenic pituitaries were comparable to those of control mice. Finally, POMV3 mice responded with a similar maximal increase of ACTH and corticosterone to a 20-min acute restraint stress. Together, these results show that hypophyseal V3 overexpression led to increased basal concentrations of corticosterone and suggest that the negative glucocorticoid feedback may be altered at the pituitary level.

Glucocorticoids play an important role in determining adipose tissue metabolism and distribution. Patients with Cushing’s syndrome or receiving corticosteroid therapy develop a reversible visceral obesity. In obese patients, although circulating concentrations of cortisol are not consistently elevated, local conversion of inactive cortisone to active cortisol in adipose tissue, catalyzed by 11-hydroxysteroid dehydrogenase type 1 (11-HSD-1), could amplify glucocorticoid signaling. We have studied, using semiquantitative in situ hybridization, 11-HSD-1 mRNA expression in the adipocyte and stromal compartments of sc abdominal adipose tissue obtained from 12 lean patients and sc abdominal and visceral adipose tissue obtained from 18 obese patients. 11-HSD-1 mRNA was expressed in adipocytes, stroma, and walls of vessels. Localization of 11-HSD-1 mRNA did not differ between lean sc and obese sc or visceral adipose tissue. 11-HSD-1 mRNA levels were significantly (P 0.0106) increased in the adipocyte compartment of sc adipose tissue obtained from obese patients as compared with nonobese ones, whereas no significant change (P 0.446) was found in the stromal compartment. In obese patients, 11-HSD-1 mRNA expression was increased (P 0.0157) in the stromal compartment of visceral compared with sc tissue, whereas no significant change (P 0.8767) was found in the adipocyte compartment. In summary, our data show that 11-HSD-1 mRNA is increased in adipose tissue from obese patients, in the abdominal sc fat in adipocytes and in the visceral fat in both adipocytes and stroma. This observation suggests that an overexpression of 11-HSD-1 may explain part of the glucocorticoid-induced metabolic disorders linked to obesity and may promote visceral fat deposition. (J Clin Endocrinol Metab 87: 2701–2705, 2002)

Objectives: To compare the feasibility, safety, and outcome of parathyroidectomy in the management of primary hyperparathyroidism (PHPT) in individuals aged 75 and older with that of those younger than 50. Design: Retrospective chart review. Setting: Department of Endocrine Surgery, Hospital Paul Desbief (Marseille, France). Participants: Individuals who underwent surgery from June 2005 to February 2013 (N = 526) had a clinical examination and laboratory and imaging assessments to diagnose PHPT. Measurements: The clinical and biochemical characteristics and surgery outcomes of individuals younger than 50 (n = 80) were compared with the characteristics and outcomes of those aged 75 and older (n = 89). Results: Most of the participants did not have any specific signs of PHPT, and the diagnosis of PHPT was established in some participants during routine clinical and laboratory examination. Nephrolithiasis and osteitis fibrosa cystica were observed only in the younger group. Urinary calcium decreased with age. Nine participants aged 75 and older did not undergo surgery (four declined, five had medical contraindications). Conventional surgery through transverse cervicotomy was used in the majority of participants. Cure rate was excellent (158/160), with few and reversible minor complications. The coexistence of thyroid lesions was significantly higher in the older (47.5%) than in the younger group (32.3%). Nodules and multinodular goiters were removed in the majority of participants during the parathyroidectomy procedure. Conclusion: With the exception of a few cases with severe associated comorbidities, parathyroidectomy is safe and curative and should be considered as first-line choice for older adult with PHPT.

The effects of ionotropic excitatory amino acids agonists on the release of vasopressin from rat hypothalamic slices were studied. Incubation with increasing doses of NMDA, kainate or AMPA decreased the release of vasopressin in a dose-dependent manner. The values of the IC50 were 1.0, 9.6, or 3.7 x 10-8 M, respectively. The inhibitory effect of the various excitatory amino acids tested was blocked by coincubation with their respective antagonists. Vasopressin secretion was stimulated to 140.3 +/- 7.6% of controls when the slices were obtained from chronically (7 days) salt-loaded rats. Addition of 1 x 10-7 M NMDA or 1 x 10-6 M kainate to the incubation medium antagonized the salt loading-induced increase in vasopressin release. Incubation with 1 x 10-4 M tetrodotoxin did not change basal vasopressin release, but it blocked the decrease in vasopressin secretion induced by 1 x 10-7 M NMDA or 1 x 10-6 M kainate or 1 x 10-6 M AMPA. Incubation with 1 x 10-5 M phaclophen (a GABAB antagonist) and 1 x 10-5 M bicuculline (a GABAA antagonist) was without effect on basal vasopressin secretion while it reversed the inhibition of vasopressin release induced by 1 x 10-7 M NMDA. Incubation with 1 x 10-6 M GABA alone decreased vasopressin secretion to 64.6 +/- 6.9% of control values. The inhibitory effect of GABA did not change when 1 x 10-7 M NMDA was added to the incubation medium. These findings demonstrate that ionotropic excitatory amino acids agonists inhibit vasopressin secretion from hypothalamic slices. They strongly suggest that this inhibitory effect is mediated through local GABAergic interneurones.

Neonatal rats were daily injected with 100 microg/kg T4 and killed at 4, 8 or 15 days. Circulating corticosterone and corticosteroid binding globulin concentrations increased in 8- and 15-day-old rats after T4 treatment. Plasma adrenocorticotropic hormone (ACTH) concentrations, pituitary ACTH content and pro-opiomelanocortin mRNA expression were unaffected in T4-treated rats. T4 treatment induced an increase in corticotropin-releasing factor (CRF) mRNA expression in the whole population of CRF synthesizing cells of the paraventricular nucleus (PVN) that became significant at day 8 and disappeared at day 15. Double labelling in situ hybridization revealed that CRF gene expression in the CRF+/arginine vasopressin (AVP)+ subpopulation was increased at days 4 and 8 and decreased at day 15. CRF immunoreactivity in the zona externa of the median eminence increased with age but was not affected by the experimental hyperthyroidism. The degree of CRF and AVP colocalization, the concentration of AVP mRNA in the parvo and magnocellular cell bodies of the PVN and the density of immunoreactive AVP in the zona interna or zona externa of the median eminence did not change after T4 treatment. Our data demonstrate that experimental hyperthyroidism accelerates the maturation of hypothalamic CRF gene expression, including in particular in the CRF+/AVP+ subpopulation, during the stress hyporesponsive period. These observations suggest that the physiological peak of plasma thyroxine that occurs between days 8-12 may participate in the maturation of hypothalamic CRF cells.

As fetal overexposure to glucocorticoids has been postulated to induce intrauterine growth retardation (IUGR) in humans, we investigated the effects of maternal 50% food restriction (FR50) in rats during the last week of gestation on the hypothalamo-pituitary adrenal (HPA) axis activity in both mothers and their fetuses. In mothers, FR50 increased both the plasma corticosterone (B) level from embryonic days 19–21 and the relative adrenal weight at term. FR50 decreased at term both the maternal plasma corticosteroid-binding globulin level and placental 11β-hydroxysteroid dehydrogenase type 2 expression. In newborns, maternal FR50 reduced body and adrenal weights, glucocorticoid and mineralocorticoid receptor expressions in the hippocampus, corticoliberin expression in the hypothalamic paraventricular nucleus, and plasma ACTH. In FR50 newborns, the plasma B level was increased at birth and decreased 2 h later. When maternal circulating B was maintained at the basal level by adrenalectomy and B supply, FR50 ...

Malignant thyroid tumors reportedly exhibit an anomaly in thyroid peroxidase (TPO) resulting in a lower affinity for monoclonal antibody 47 (mAb 47) in immunohistochemistry studies. The purpose of the present study was to compare TPO immunostaining in normal, benign, and malignant thyroid tissue with expression of mRNA sequences in four exons of the molecule including the epitope of mAb 47. TPO immunostaining was performed using mAb 47 and a polyclonal antibody (pAb). Messenger RNA expression was investigated by in situ hybridization using probes specific for mRNA sequences in exons 2, 12 (epitope of mAb), 15, and 17. As expected, pAb immunostaining was significantly positive on all benign tumors and 50% of carcinomas. With mAb 47, little or no immunostaining was observed in 16 of 17 carcinomas while significantly positive immunostaining was found in normal tissue and benign tumors. In situ hybridization showed a decrease and heterogeneity in the expression of all mRNA sequences in carcinomas as compared to normal tissue and benign tumors. Unlike the other three probes, the probe specific for exon 12 hybridized strongly with benign tumors but poorly with most carcinomas. Poor hybridization was usually correlated with defective mAb 47 immunostaining. These results confirm that TPO is expressed in thyroid carcinomas but in smaller amounts than in normal tissue and benign tumors. In malignant tumors, qualitative changes in TPO may also impede mAb 47 immunostaining. In situ hybridization showed a concomitant decrease in the corresponding TPO mRNA sequence. These changes could be due to abnormalities in the maturation of TPO mRNA leading to a different splicing variant.

We examined the expression of functional growth hormone secretagogue receptors (GHS-R) in a series of 30 human pituitary adenomas-six secreting GH, three GH-PRL, six prolactin (PRL), five adrenocorticotrophic hormone (ACTH), one thyroid stimulating hormone (TSH), four gonadotroph and five non-secreting adenomas. By reverse transcriptase polymerase chain reaction (RT-PCR), the coexpression of the two GHS-R isoforms (Ia and Ib) was found in all the GH-, GH-PRL- and PRL-secreting adenomas, and only in two out of three corticotroph, two out of four gonadotroph and one out of five non-secreting tumours. They were absent in the TSH-secreting adenoma. The PCR products of GHS-R Ia and Ib were identical in size to those from two normal pituitaries. PCR cloning and sequencing of isoforms performed in two somatotroph adenomas revealed only two single, silent base mutations. Triple in-situ hybridization showed colocalization of GHS-R mRNA with messengers of GH and PRL, conjointly or separately, in individual cells of somatotroph, mammosomatotroph, and lactotroph adenomas. The presence of GHS-R mRNA in cells expressing PRL mRNA is emphasized. In cultured cells from six somatotroph and two mammosomatotroph adenomas, the powerful GHS MK-0677 stimulated GH release in a dose-dependent manner, with maximal effect at 6 h. Contrarily, when GHRH was applied, only three somatotrophs and two mamosomatotrophs were stimulated. In the two mammosomatotrophs, the PRL response to MK-0677 and to GHRH was similar to the GH response. An homologous desensitization of the GHS-R and the GHRH receptor was observed 24 h after a first stimulation by a single dose of the corresponding agonist. Heterologous desensitization was not observed. Interestingly, MK-0677 also stimulated, in a dose-dependent way, the hormone release of cells from all tested lactotroph and corticotroph adenomas. The existence of a functional expression of GHS-R in somatotroph, mammosomatotroph, lactotroph and corticotroph adenomas rises the question of the role played by GHS-R in pituitary adenomas, particularly those not engaged in GH secretion.

A number of putative transcription factors described in the pituitary have been implicated as key elements in the processes that direct pituitary development. Three recently described proteins, Ptx1, Ptx2, and Ptx3, define a new family of transcription factors, the Ptx subfamily, within the paired-like class of homeodomain factors. In mice, Ptx1 and Ptx2 gene expression has been detected in the area of the pituitary primordium and is maintained throughout development in Rathke pouch and adult pituitary. In the present study, the expression of the Ptx1, Ptx2, and Ptx3 genes was characterized in the normal human pituitary and in the different types of human pituitary adenomas. Although no Ptx3 gene expression could be detected in these tissues, Ptx1 presented with a quite ubiquitous pattern of distribution, being expressed at quite constant levels in normal tissues and in all 60 pituitary tumors analyzed. The pattern of expression of the Ptx2 gene among the different subsets of pituitary adenomas was even more varied. No Ptx2 expression could be detected in corticotroph tumors. In contrast, high levels of Ptx2 messenger ribonucleic acid were measured in the gonadotroph tumors, although no specific correlation to other markers of the gonadotroph lineage differentiation, such as alphaGsu, LHbeta, or FSHbeta, could be evidenced. Finally, Ptx2 was also expressed in pure lactotroph adenomas and not in somatotroph adenomas. Ptx2 is, therefore, the first paired homeodomain pituitary transcription factor differentially expressed in these two lineages, which derive from a common precursor. These results support a role for Ptx2 in the terminal differentiation of somatotroph and lactotroph cell phenotypes.

OBJECTIIVES To determine the endocrine response to surgical stress in a homogeneous population of 36 women with microprolactinomas, particularly to evaluate the effect of the sharp decrease in plasma prolactin on stress induced hormonal secretion. In addition, the effects of exogenous opiates on prolactin secretion were studied. METHODS The plasma kinetics of cortisol, prolactin, ACTH, GH, and β-endorphin like immunoreactivity (β-ELI) were analysed by including patients operated on with strict anaesthetic and surgical protocols, and by sampling blood every 10 minutes, starting at premedication up to 3 hours after induction. RESULTS AND CONCLUSION S ( a ) Surgical stress or opiate administration did not induce prolactin release in patients with microprolactinoma. ( b ) The dramatic decrease in prolactin concentrations have apparently no effect on the release of other hormones involved in stress. ( c ) The existence of an early GH peak, independently of any surgical procedure, strongly suggests that GH is released by surgical stress whereas β-endorphin is secreted in response to pain. Thus GH may be a useful marker of surgical stress.

We present a specific and sensitive method for simultaneous detection of three mRNA species in individual neurons. The method relies on the use of riboprobes labeled with [35S]-UTP, digoxigenin-UTP, or biotin-UTP. The nonradioactive probes were sequentially revealed by incubation with anti-digoxigenin immunoglobulins or streptavidin conjugated to peroxidase, followed by the use of fluorochrome-labeled tyramides as peroxidase substrates. The radioactive probe was revealed by conventional autoradiography. There was no interaction among the different probes or the various detection systems. We demonstrate the use of this method by illustrating on laser scanning confocal microscopy the co-localization of the mRNAs coding for corticotropin-releasing factor (CRF), arginine vasopressin (AVP), or peptidylglycine α-amidating monooxygenase (PAM) in rat hypothalamic paraventricular nucleus (PVN) and its modulation by endogenous glucocorticoids. Our results suggest that this method could be used not only to study the regulation of the hypothalamo-pituitary-adrenal axis but also in various models in which mRNAs are present at low concentrations.