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Anna Malczewska Department of Endocrinology and Neuroendocrine Tumours, Medical University of Silesia, Katowice, Poland

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Kjell Oberg Department of Endocrine Oncology, University Hospital, Uppsala, Sweden

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Beata Kos-Kudla Department of Endocrinology and Neuroendocrine Tumours, Medical University of Silesia, Katowice, Poland

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available ( 12 ). These include enzyme linked immunosorbent assay (ELISA), IRMA, or RIA, and more recently an immunofluorescent assay based on Time-Resolved Amplified Cryptate Emission (TRACE) ( 9 ). The variety of assays reflect the presence of several CgA

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Justyna Modrzynska Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark

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Christine F Klein Department of Cardiology, Herlev Gentofte Hospital, Herlev, Denmark

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Kasper Iversen Department of Clinical Medicine, Herlev Gentofte Hospital, Herlev, Denmark

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Henning Bundgaard Department of Cardiology, Rigshospitalet, University of Copenhagen, Copenhagen, Denmark

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Bolette Hartmann Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
Novo Nordisk Foundation Center for Basic Metabolic Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark

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Maike Mose Department of Endocrinology and Internal Medicine, Aarhus University Hospital, Aarhus, Denmark

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Nikolaj Rittig Steno Diabetes Center Aarhus, Aarhus University Hospital, Aarhus, Denmark

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Niels Møller Department of Clinical Medicine, Aarhus University Hospital, Aarhus, Denmark

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Jens J Holst Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
Novo Nordisk Foundation Center for Basic Metabolic Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark

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Nicolai J Wewer Albrechtsen Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
Department of Clinical Biochemistry, Rigshospitalet, University of Copenhagen, Copenhagen, Denmark

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measurement of glucagon, GLP-1 and CRP Plasma concentration of glucagon was determined using a sandwich ELISA assay employing N- and C-terminus glucagon-specific antibody. This assay has previously been validated ( 19 ). Plasma concentration of total GLP-1

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Maximilian Bielohuby Sanofi-Aventis Deutschland GmbH, R&D, Industriepark Höchst, Frankfurt, Germany

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Martin Bidlingmaier Endocrine Research Laboratories, Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, Munich, Germany

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Uwe Schwahn Sanofi-Aventis Deutschland GmbH, R&D, Industriepark Höchst, Frankfurt, Germany

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research purposes in laboratory rodents. The replacement of radioactivity by an enzymatic label in 1971 by Engvall and Perlmann (ELISA) ( 5 ) as well as by van Weemen and Schuurs (EIA) ( 6 ), and the associated increased handling convenience led to a boost

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Erik Rösner Institute of Pharmacology and Toxicology, Jena University Hospital, Jena, Germany

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Daniel Kaemmerer Department of General and Visceral Surgery, Zentralklinik Bad Berka, Bad Berka, Germany

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Elisa Neubauer Institute of Pharmacology and Toxicology, Jena University Hospital, Jena, Germany

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Jörg Sänger Laboratory of Pathology and Cytology Bad Berka, Bad Berka, Germany

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Amelie Lupp Institute of Pharmacology and Toxicology, Jena University Hospital, Jena, Germany

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and designed the experiments. Daniel Kaemmerer and Jörg Sänger provided the tumour samples. Daniel Kaemmerer provided the clinical data. Erik Rösner, Elisa Neubauer and Amelie Lupp performed the experiments. Erik Rösner and Amelie Lupp analysed the

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Fabyan Esberard de Lima Beltrão Lauro Wanderley University Hospital, Federal University of Paraíba, João Pessoa, Paraíba, Brazil
Postgraduate Program in Nutritional Sciences, Department of Nutrition, Center for Health Sciences, Federal University of Paraíba, João Pessoa, Paraíba, Brazil
University Centre of João Pessoa (UNIPE), João Pessoa, Paraíba, Brazil

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Daniele Carvalhal de Almeida Beltrão University Centre of João Pessoa (UNIPE), João Pessoa, Paraíba, Brazil
Postgraduate Program in Cognitive Neuroscience and Behavior, Center for Health Sciences, Federal University of Paraíba, João Pessoa, Paraíba, Brazil

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Giulia Carvalhal Center for Biological and Health Sciences, Federal University of Campina Grande, Campina Grande, Paraíba, Brazil

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Fabyo Napoleão de Lima Beltrão Department of Medicine, Faculty of Medical Sciences, João Pessoa, Brazil

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Igor Motta de Aquino Metropolitan Hospital Dom José Maria Pires, Santa Rita, Paraíba, Brazil

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Thaíse da Silva Brito New Hope Medical School – FAMENE, João Pessoa, Paraíba, Brazil

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Barbara Costa Paulino Postgraduate Program in Nutritional Sciences, Department of Nutrition, Center for Health Sciences, Federal University of Paraíba, João Pessoa, Paraíba, Brazil

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Elisa Aires Postgraduate Program in Interactive Processes of Organs and Systems, Health & Science Institute, Federal University of Bahia, Salvador, Bahia, Brazil

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Diana Viegas Internal Medicine Department, rede UniFTC, Salvador, Bahia, Brazil

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Fabio Hecht The Institute of Biophysics Carlos Chagas Filho, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil

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Bruno Halpern Weight Control Centre, Hospital 9 de Julho, São Paulo, São Paulo, Brazil

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Liana Clebia De Morais Pordeus Postgraduate Program in Cognitive Neuroscience and Behavior, Center for Health Sciences, Federal University of Paraíba, João Pessoa, Paraíba, Brazil

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Maria da Conceição Rodrigues Gonçalves Postgraduate Program in Nutritional Sciences, Department of Nutrition, Center for Health Sciences, Federal University of Paraíba, João Pessoa, Paraíba, Brazil

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Helton Estrela Ramos Postgraduate Program in Interactive Processes of Organs and Systems, Health & Science Institute, Federal University of Bahia, Salvador, Bahia, Brazil
Department of Biorregulation, Health Sciences Institute, Federal University of Bahia, Bahia, Brazil
Postgraduate Program in Medicine and Health, Medical School of Medicine, Federal University of Bahia, Salvador, Bahia, Brazil

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-reactive protein (hs-CRP), and lactate dehydrogenase (LDH). Leptin was measured with an ELISA (DiaSorin, Inc., Stillwater, Minnesota, USA). The method used in the other exams was automated chemiluminescence (MAGLUMI-2000-PLUS; Shenzhen New Industries Biomedical

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Lesley A Hill Departments of Cellular and Physiological Sciences and Obstetrics and Gynaecology, The University of British Columbia, Vancouver, British Columbia, Canada

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Zeynep Sumer-Bayraktar School of Life and Environmental Science, Charles Perkins Centre, The University of Sydney, Sydney, New South Wales, Australia

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John G Lewis Canterbury Health Laboratories, Christchurch, New Zealand

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Eva Morava Mayo Clinic, Department of Clinical Genomics, CIM, Rochester, Minnesota, USA

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Morten Thaysen-Andersen Department of Molecular Sciences, Macquarie University, Sydney, New South Wales, Australia

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Geoffrey L Hammond Departments of Cellular and Physiological Sciences and Obstetrics and Gynaecology, The University of British Columbia, Vancouver, British Columbia, Canada

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frequency of N -glycosylation abnormalities associated with disease ( 14 , 15 ). ELISAs have been developed for human CBG measurements in biological fluids utilizing monoclonal antibodies that recognize specific epitopes on the surface of the natively

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Filippo Ceccato Endocrinology Unit, Department of Medicine DIMED, University-Hospital of Padova, Padova, Italy

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Elisa Selmin Endocrinology Unit, Department of Medicine DIMED, University-Hospital of Padova, Padova, Italy

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Chiara Sabbadin Endocrinology Unit, Department of Medicine DIMED, University-Hospital of Padova, Padova, Italy

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Miriam Dalla Costa Endocrinology Unit, Department of Medicine DIMED, University-Hospital of Padova, Padova, Italy

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Giorgia Antonelli Laboratory Medicine, Department of Medicine DIMED, University-Hospital of Padova, Padova, Italy

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Mario Plebani Laboratory Medicine, Department of Medicine DIMED, University-Hospital of Padova, Padova, Italy

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Mattia Barbot Endocrinology Unit, Department of Medicine DIMED, University-Hospital of Padova, Padova, Italy

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Corrado Betterle Endocrinology Unit, Department of Medicine DIMED, University-Hospital of Padova, Padova, Italy

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Marco Boscaro Endocrinology Unit, Department of Medicine DIMED, University-Hospital of Padova, Padova, Italy

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Carla Scaroni Endocrinology Unit, Department of Medicine DIMED, University-Hospital of Padova, Padova, Italy

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Introduction and Aim

The purpose of replacement therapy in adrenal insufficiency (AI) is mimicking endogenous cortisol levels as closely as possible: dual release hydrocortisone (DR-HC) has been introduced to replicate the circadian cortisol rhythm. Multiple daily saliva collections could be used to assess the cortisol rhythm during real life: our aim was to study the salivary cortisol profile in AI.

Materials and Methods

We prospectively evaluated, in an observational study, 18 adult outpatients with AI (11 primary and 7 secondary AI), switched from conventional treatment (conv-HC, 25 mg/day) to the same dose of DR-HC. We collected six samples of saliva in a day, measuring cortisol (F) and cortisone (E) with LC-MS/MS. Forty-three matched healthy subjects served as controls.

Results

F levels were similar in the morning (and higher than controls) in patients treated with conv-HC or DR-HC; otherwise F levels and exposure were lower in the afternoon and evening in patients with DR-HC, achieving a cortisol profile closer to healthy controls. Daily cortisol exposure, measured with area under the curve, was lower with DR-HC. Morning F and E presented sensitivity and specificity >90% to diagnose AI (respectively threshold of 3 and 9.45 nmol/L). Total cholesterol and HbA1c levels reduced with DR-HC.

Conclusions

Salivary cortisol daily curve could be used as a new tool to assess the cortisol profiles in patients treated with conv-HC and DR-HC. A lower daily cortisol exposure was achieved with DR-HC (despite the same HC dose), especially in the afternoon-evening.

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Nella Augusta Greggio Endocrinology and Adolescence Unit, Department of Woman and Child Health, University of Padova, Padova, Italy

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Elisa Rossi CINECA – Interuniversity Consortium (Health Service), Bologna, Italy

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Silvia Calabria CORE srl – Collaborative Outcome Research, Bologna, Italy

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Alice Meneghin Endocrinology and Adolescence Unit, Department of Woman and Child Health, University of Padova, Padova, Italy

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Joaquin Gutierrez de Rubalcava Endocrinology and Adolescence Unit, Department of Woman and Child Health, University of Padova, Padova, Italy

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Carlo Piccinni CORE srl – Collaborative Outcome Research, Bologna, Italy

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Antonella Pedrini CORE srl – Collaborative Outcome Research, Bologna, Italy

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Objective

To estimate the prevalence of subclinical hypothyroidism (SH) among children, by using levothyroxine low dosage as disease proxy, and to describe prescription pattern.

Design

An historical cohort study was performed through administrative databases of 12 Italian Local Health Units covering 3,079,141 inhabitants. A cohort of children (aged 0–13 years) was selected in the period 2001–2014. A subgroup of new users (aged 0–9 years) was identified and followed up for 5 years.

Methods

The prevalence was provided as mean value of the whole period, as annual trend, by patient gender and age. Demographic details, information on levothyroxine dosage, comorbidities and co-medications were provided. Therapy duration and medication persistence were evaluated among new users.

Results

644 children treated with levothyroxine low dosage was selected, with a mean annual prevalence of 0.20 per 1000 children. The temporal trend of prevalence was stable, with a slight reduction in the 2005–2008. Prevalence by age showed an increase after 10 years. Patients were treated with an average annual dose of 4290 µg/year and 66.9% of children were affected by comorbidities. Among 197 new users, 62.9% received therapy only for one year, whereas out of those treated two or more years, 89.0% resulted persistent to the therapy.

Conclusions

This study provides real-world epidemiology of SH among children, and it depicts the clinical and therapeutic characteristics of these subjects. Its findings showed that the SH treatment of this disorder was widely variable, also due to lack of evidence concerning paediatric population.

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Filippo Ceccato Department of Medicine DIMED, University of Padova, Padova, Italy
Endocrine Disease Unit, University-Hospital of Padova, Padova, Italy
Department of Neuroscience DNS, University of Padova, Padova, Italy

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Elisa Selmin Department of Medicine DIMED, University of Padova, Padova, Italy

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Giorgia Antonelli Department of Medicine DIMED, University of Padova, Padova, Italy
Laboratory Medicine, University-Hospital of Padova, Padova, Italy

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Mattia Barbot Department of Medicine DIMED, University of Padova, Padova, Italy
Endocrine Disease Unit, University-Hospital of Padova, Padova, Italy
Department of Neuroscience DNS, University of Padova, Padova, Italy

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Andrea Daniele Department of Medicine DIMED, University of Padova, Padova, Italy

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Marco Boscaro Department of Medicine DIMED, University of Padova, Padova, Italy

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Mario Plebani Department of Medicine DIMED, University of Padova, Padova, Italy
Laboratory Medicine, University-Hospital of Padova, Padova, Italy

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Carla Scaroni Department of Medicine DIMED, University of Padova, Padova, Italy
Endocrine Disease Unit, University-Hospital of Padova, Padova, Italy

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Context

The low-dose short synacthen test (LDSST) is recommended for patients with suspected central adrenal insufficiency (AI) if their basal serum cortisol (F) levels are not indicative of an intact hypothalamic–pituitary–adrenal (HPA) axis.

Objective

To evaluate diagnostic threshold for salivary F before and 30 min after administering 1 μg of synacthen, performed before 09:30 h.

Design

A cross-sectional study from 2014 to 2020.

Setting

A tertiary referral university hospital.

Patients

In this study, 174 patients with suspected AI, 37 with central AI and 137 adrenal sufficient (AS), were included.

Main outcome measure

The diagnostic accuracy (sensitivity (SE), specificity (SP)) of serum and salivary F levels measured, respectively, by chemiluminescence immunoassay and liquid chromatography-tandem mass spectrometry.

Results

Low basal serum or salivary F levels could predict AI. For the LDSST, the best ROC-calculated threshold for serum F to differentiate AI from AS was 427 nmol/L (SE 79%, SP 89%), serum F > 500 nmol/L reached SP 100%. A salivary F peak > 12.1 nmol/L after administering synacthen reached SE 95% and SP 84% for diagnosing central AI, indicating a conclusive reduction in the likelihood of AI. This ROC-calculated threshold for salivary F was similar to the 2.5th percentile of patients with a normal HPA axis, so it was considered sufficient to exclude AI. Considering AS those patients with salivary F > 12.1 nmol/L after LDSST, we could avoid unnecessary glucocorticoid treatment: 99/150 subjects (66%) had an inadequate serum F peak after synacthen, but salivary F was >12.1 nmol/L in 79 cases, who could, therefore, be considered AS.

Conclusions

Salivary F levels > 12.1 nmol/L after synacthen administration can indicate an intact HPA axis in patients with an incomplete serum F response, avoiding the need to start glucocorticoid replacement treatment.

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Soraya Puglisi Internal Medicine 1, Department of Clinical and Biological Sciences, University of Turin, Turin, Italy

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Paola Perotti Internal Medicine 1, Department of Clinical and Biological Sciences, University of Turin, Turin, Italy

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Mattia Barbot Endocrinology Unit, Department of Medicine DIMED, University of Padua, Padua, Italy

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Paolo Cosio Internal Medicine 1, Department of Clinical and Biological Sciences, University of Turin, Turin, Italy

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Carla Scaroni Endocrinology Unit, Department of Medicine DIMED, University of Padua, Padua, Italy

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Antonio Stigliano Endocrinology Unit, Department of Clinical and Molecular Medicine, Sant’Andrea Hospital University of Rome, Rome, Italy

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Pina Lardo Endocrinology Unit, Department of Clinical and Molecular Medicine, Sant’Andrea Hospital University of Rome, Rome, Italy

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Valentina Morelli Endocrinology Unit, Department of Clinical Sciences and Community Health, University of Milan and Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico, Milano, Italy

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Elisa Polledri Department of Clinical Sciences and Community Health, Laboratory of Toxicology, University of Milan and Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico, Milan, Italy

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Iacopo Chiodini Endocrinology Unit, Department of Clinical Sciences and Community Health, University of Milan and Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico, Milano, Italy

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Giuseppe Reimondo Internal Medicine 1, Department of Clinical and Biological Sciences, University of Turin, Turin, Italy

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Anna Pia Internal Medicine 1, Department of Clinical and Biological Sciences, University of Turin, Turin, Italy

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Massimo Terzolo Internal Medicine 1, Department of Clinical and Biological Sciences, University of Turin, Turin, Italy

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Objective

Metyrapone has been approved for the treatment of patients with Cushing’s syndrome (CS), but only few retrospective clinical studies are available. The aim of our study was the prospective assessment of metyrapone as pre-operative treatment.

Design and methods

Before adrenalectomy, seven patients with ACTH-independent CS due to adrenal adenoma were prospectively treated with metyrapone for 3 months in three tertiary academic centers, with endocrine work-up and clinical evaluation at screening and at predefined evaluation time points (Days 14, 31, 48, 65, 82).

Results

In all patients, UFC levels decreased up to normal range from baseline to Day 82 (609 (188–1476) vs 69 (28–152) nmol/24 h, P < 0.02), with a reduction of serum and salivary cortisol levels, and no significant increase of plasma ACTH and serum DHEAS levels. Clinical improvement was reported on quality of life (+16.7 (+4.2; +52.00) points, P < 0.04) and pressure control (systolic pressure, −25 (−52; −10) mmHg, P < 0.01; diastolic pressure, −16 (−50; +2 mmHg), P < 0.03). No significant change in weight, electrolytes, glycemic and lipid profile was reported. Although in women a significant increase of testosterone and androstenedione was reported, no worsening of clinical hyperandrogenism was observed. All drug-related adverse events (nausea, fatigue, low grade fever, edema of lower limbs and facial rash) were grade 1 or 2 and generally transient.

Conclusions

This prospective pilot study demonstrated that metyrapone is effective in normalizing biochemical and clinical parameters in patients with CS due to adrenal adenoma before surgical intervention, with minimal side effects.

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