Background: This study aimed to compare a chemiluminescence immunoassay (CLIA; Siemens Atellica IM 1600) with an automated liquid chromatography–tandem mass spectrometry (LC–MS/MS) analyser (Thermo Scientific Cascadion SM) for serum 25-hydroxyvitamin D [25(OH)D] measurement in unselected outpatients, and its impact on clinical classification. Methods: 25(OH)D was measured on 1064 paired serum samples (828 women, 236 men; age 3–111 years); both platforms participated in the Vitamin D Standardisation Program (VDSP). Agreement was assessed by the Wilcoxon signed-rank test, Passing–Bablok regression, Bland–Altman analysis on log-transformed values, Breusch–Pagan testing for heteroscedasticity, consistency-based intraclass correlation coefficient (ICC), Lin’s concordance correlation coefficient, and Cohen’s kappa with McNemar’s test on six- and two-category clinical classifications. Results: Methods differed significantly (p<2.2×10−16). Passing–Bablok regression showed proportional bias (slope 1.33; 95% confidence interval [CI], 1.28–1.38; LC–MS/MS higher). Consistency ICC was 0.928 (95% CI, 0.919–0.936). Log-Bland–Altman bias was 35.5% (limits of agreement, −18.3% to +124.5%; heteroscedasticity, p=5.18×10−11). Cohen’s kappa was 0.32 unweighted and 0.72 weighted on six clinical categories, and 0.56 on a two-class scheme (McNemar p<2.2×10−16). In discordant pairs, CLIA classified deficiency 118-fold more often than LC–MS/MS. Conclusions: In our study, despite preserved rank ordering, a non-linear concentration-dependent bias and different limits of agreement caused systematic differences in the classification of vitamin D deficiency. Adoption of automated LC–MS/MS can substantially reclassify outpatients toward sufficiency, with implications for supplementation and laboratory harmonisation.

Among the various environmental pollutants, dioxin, a highly toxic and widely used compound, is associated with numerous adverse health effects, including a potentially toxic multigenerational effect. Understanding the mechanisms by which dioxin exposure can affect sperm epigenetics is critical to comprehending the potential consequences for offspring health and development. This study investigates the possible association between weighted epimutations, hypothesized as markers of epigenetic drift, and dioxin exposure in sperm tissues. We used a public online methylation dataset consisting of 37 participants: 26 Vietnam veterans exposed to Agent Orange, an herbicide contaminated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and 11 individuals not directly exposed to TCDD but whose serum dioxin levels are equivalent to the background. In our study, conducted at the gene level, 437 epimutated genes were identified as significantly associated with each single-digit increase in serum dioxin levels. We found no significant association between the rise in total epimutation load and serum dioxin levels. The pathway analysis performed on the genes reveals biological processes mainly related to changes in embryonic morphology, development, and reproduction. Results from our current study suggest the importance of further investigations on the consequences of dioxin exposure in humans with specific reference to germinal tissue and related heredity.

Context In 2005, a nationwide program of iodine prophylaxis on a voluntary basis was implemented in Italy by law. However, recent data on iodine status are lacking. Objective The aim of this study was to evaluate efficiency, effectiveness, and possible adverse effects (increased occurrence of thyroid autoimmunity and hyperthyroidism) of the Italian iodine prophylaxis program. Methods From 2015 to 2019, a nationwide survey was performed. The use of iodized salt was evaluated in a sample of 164 593 adults and in 998 school canteens. A sample of 4233 schoolchildren (aged 11-13 years) was recruited to assess urinary iodine concentration, prevalence of goiter, and thyroid hypoechogenicity on ultrasound, with the latter being an indirect indicator of thyroid autoimmunity. Neonatal TSH values of 197 677 infants screened in regions representative of Northern, Central, and Southern Italy were analyzed to investigate the percentage of TSH values >5.0 mIU/L. Data on methimazole prescriptions were analyzed as indirect indicators of new cases of hyperthyroidism. Results The prevalence of the use of iodized salt was 71.5% in adult population and 78% in school canteens. A median urinary iodine concentration of 124 μg/L, a prevalence of goiter of 2.2%, and a prevalence of thyroid hypoechogenicity of 5.7% were observed in schoolchildren. The percentage of neonatal TSH values >5.0 mIU/L resulted still higher (5.1%) than the World Health Organization threshold of 3.0%, whereas the prescriptions of methimazole showed a reduction of 13.5%. Conclusion Fifteen years of iodine prophylaxis have led to iodine sufficiency in Italy, although there still is concern about iodine nutritional status during pregnancy.

In Italia il processo di diagnosi e cura del paziente in età neonatale e pediatrica con ipotiroidismo congenito (IC) viene favorito dalla disponibilità di un documento, approvato nel 2018 dalla Società Italiana di Endocrinologia e Diabetologia Pediatrica (SIEDP), che definisce i principi base del percorso diagnostico-terapeutico (PDTA) del neonato e del bambino con IC. Il documento è concepito per rispondere all’esigenza italiana di un percorso applicato su tutto il territorio nazionale con il duplice obiettivo di: 1) implementare il processo di diagnosi e cura del paziente con IC in aderenza alle linee guida internazionali; e 2) individuare modalità operative che consentano di migliorare la qualità dell’assistenza. In particolare, il PDTA offre un modello operativo che favorisce la sinergia fra strutture assistenziali ospedaliere e territoriali coinvolte nella gestione del paziente con IC, rendendo la sua attuazione applicabile nei contesti locali.

Simona De Angelis1 · Daniela Rotondi1 · Enzo Gilardi1 · Paolo Stacchini2 · Augusto Alberto Pastorelli2 · Angela Sorbo2 · Marilena D’Amato2 · Anna Chiara Turco2 · Emanuela Medda3 · Roberto Da Cas4 · Sebastiano Andò5 · Daniela Bonofiglio5 · Marcello Bagnasco6 · Maurizio Gasperi7 · Domenico Meringolo8 · Caterina Mian9 · Efisio Puxeddu10 · Concetto Regalbuto11 · Mariacarla Moleti12 · Augusto Taccaliti13 · Salvatore Ulisse14 · Massimo Tonacchera15 · Maria Laura Tanda16 · Francesco Boi17 · Valeria Ruggiero17 · Stefano Mariotti17 · Carlo Corbetta18 · Renzo Ciatti19 · Elisabetta Tarsi19 · Vera Stoppioni19 · Nicola Perrotti20 · Onorina Marasco21 · Giovanna Scozzafava21 · Marta Camilot22 · Francesca Teofoli22 · Francesca Righetti23 · Antonio Dimida15 · Giuseppe Plutino24 · Elena Carrano24 · Roberto Copparoni24 · Marco Gabbianelli1 · Paolo Vitti15 · Antonella Olivieri1

Context Analysis of a 2-screen program for congenital hypothyroidism (CH) was performed using differential dried-blood spot thyrotropin (bTSH) cutoffs of 10 mU/L at first screening (all infants) and 5 mU/L at second screening (selected infants). Objectives This work aimed to characterize CH infants identified by the second screening and compare infants with bTSH of 5.0 to 9.9 and 10 mU/L or greater on second screening. Design and Patients Maternal and neonatal clinical features were retrospectively analyzed for 119 CH babies detected on the second screen in the Lombardy region of Italy, 2007 to 2014. Results Fifty-two (43.7%) of the 119 CH neonates showed bTSH values ranging from 5.0 to 9.9 mU/L at the second screening (low bTSH group) and 67 (56.3%) bTSH of 10.0 mU/L or greater (high bTSH group). The frequency of thyroid dysgenesis and eutopic gland was similar in both groups, as was the frequency of permanent and transient CH. Moreover, a high frequency of extrathyroidal malformations was found in both groups. The percentage of preterm infants (57.7% vs 23.9%, P < .001) and infants admitted to the neonatal intensive care unit (50.0% vs 17.9%, P < .001) was significantly higher in the low vs the high bTSH group. In addition, maternal treatment with glucocorticoids in pregnancy was significantly more frequent in the low bTSH group than in the high bTSH group (11.5% vs 1.5%, P = .042), as well as maternal hypothyroidism and/or goiter (26.9% vs 10.4%, P = .036). Conclusions This study has demonstrated that a lower TSH cutoff at the second screening can detect additional cases of CH and that a second bTSH cutoff of 5.0 mU/L is appropriate for identifying preterm newborns and babies with associated risk factors.

A large number of people in the world need to use a wheelchair because of different disabilities. Driving a wheelchair requires complex physical and cognitive abilities which need to be trained. Virtual training helps users acquire driving skills in a safe environment. The aim of this paper is to describe and technically validate simulation models for both manual (MW) and powered wheelchairs (PW) based on immersive virtual reality CAVE (VR). As VR system, the Gait Real-time Analysis Interactive Lab (GRAIL) was used, a CAVE equipped with a motion platform with two degrees of freedom and an optoelectronic motion capture system. A real wheelchair was positioned onto the motion platform with rear wheels free to turn in MW modality, and a commercial joystick was installed on an armrest to simulate the PW modality. Passive markers were used to track the wheel rotation, the joystick and the user hand motion. Custom D-flow applications were developed to manage virtual scene response to user actions. Overground tests, based on single wheel rotation, were performed to verify the simulation model reliability. Quantitative results demonstrated that the MW simulator kinematics was consistent with a real wheelchair overground in the absence of wheel slip and inertia (median error for MW 0.40 °, no systematic bias p = 0.943, high correlation rho > 0.999, p < 0.01). The proposed solution is flexible and adaptable to different wheelchairs, joysticks and optoelectronic systems. The main limitation is the absence of force feedback. Nevertheless, it is a reliable prototype that can be used to validate new virtual scenarios as well as for wheelchair training. The next steps include the system validation with real end users and assessment of the simulator effectiveness as a training tool.

Introduction ECHS1 encodes for short-chain enoyl-CoA hydratase, a key component in b-oxidation. This enzyme is also involved in the isoleucine and valine catabolic pathways. The literature contains reports of scattered cases of ECHS1 mutation, which show a wide clinical spectrum of presentation. Despite that the clinical spectrum of the disease has not been defined so far due to the absence of previous systematic reviews and descriptions of large series of patients. Methods We performed a systematic literature review of so far reported ECHS1 mutated patients and we reported two additional cases. We pointed out clinical and neuroradiological features of all patients. Results 45 patients were included in the analysis. Based on clinical and neuroradiological feature we were able to distinguish four main phenotypes of ECHS1deficiency: a severe neonatal presentation with a rapid and fatal course and significant white matter abnormalities; a severe infantile variant with slower neurological deterioration, developmental delay, pyramidal and extrapyramidal signs, optic atrophy, feeding difficulties, and degeneration of the deep gray nuclei; a slowly progressive infantile form, qualitatively similar to the previous phenotype, but less severe with mainly basal ganglia involvement; and a final phenotype, present in only few cases, characterized by paroxysmal exercise-induced dystonic attacks, normal neurological examination between these episodes, and isolated pallidal degeneration on MRI. Interpretation ECHS1 mutations cause metabolic encephalopathy with a wide range of clinical presentations that can be grouped into four main phenotypes, each with a distinct profile in terms of severity on clinical presentation, disease course and MRI involvement.

(1) Background: Diagnostic testing for cystic fibrosis (CF) is based on a sweat chloride test (SCT) considering the appropriate signs and symptoms of the disease and results of a gene mutation analysis. In 2014, the Istituto Superiore di Sanità (ISS) established a pilot Italian external quality assessment program for CF SCT (Italian EQA-SCT), which is now a third party service carried out by the ISS. (2) Methods: The ongoing scheme is prospective, enrollment is voluntary, and the payment of a fee is required. Results are shared through a dedicated web-facility. Assessment covers the analysis, interpretation, and reporting of results. (3) Results: Thirteen, fifteen, sixteen, and fifteen different laboratories, respectively, participated from 2015 to 2016 and from 2018 to 2019 in the Italian EQA-SCT scheme. Eleven different laboratories participated each year in all four rounds of the Italian EQA-SCT. (4) Conclusions: The overall results obtained from the laboratories participating constantly clearly show that their qualitative and quantitative performance improved significantly. This is due to the opportunity-after receiving the EQA results-to constantly review their performance and address any inconsistencies. We firmly believe that participation in the EQA program will improve the quality of participating laboratories and that EQA participation should become mandatory as a fundamental requirement for laboratory accreditation.

7 ES P E Poster presented at: Congenital hypothyroidism (CH) with delayed TSH elevation: the importance of the second-screening strategy and the evolution of CH in preterm infants Silvana Caiulo1, Maria Cristina Vigone1, Antonella Olivieri2, Marianna di Frenna1, Gaia Vincenzi1, Graziano Barera1, Carlo Corbetta3, Giovanna Weber1 1 Vita-Salute San Raffaele University Pediatric Department San Raffaele Hospital, Milan, Italy. 2 Metabolism and Endocrinology Unit, Department of Cardiovascular, Dysmetabolic and Ageing-associated Diseases, National Institute of Health, Rome, Italy. 3 Regional Reference Laboratory for Neonatal Screening, Children Hospital "V. Buzzi", ASST Fatebenefratelli Sacco, Milan, Italy

Searching for mutations in the cystic fibrosis transmembrane conductance regulator gene (CFTR) is a key step in the diagnosis of and neonatal and carrier screening for cystic fibrosis (CF), and it has implications for prognosis and personalized therapy. The large number of mutations and genetic and phenotypic variability make this search a complex task. Herein, we developed, validated, and tested a laboratory assay for an extended search for mutations in CFTR using a next-generation sequencing based method, with a panel of 188 CFTR mutations customized for the Italian population. Overall, 1426 dried blood spots from neonatal screening, 402 genomic DNA samples from various origins, and 1138 genomic DNA samples from patients with CF were analyzed. The assay showed excellent analytical and diagnostic operative characteristics. We identified and experimentally validated 159 (of 188) CFTR mutations. The assay achieved detection rates of 95.0% and 95.6% in two large-scale case series of CF patients from central and northern Italy, respectively. These detection rates are among the highest reported so far with a genetic test for CF based on a mutation panel. This assay appears to be well suited for diagnostics, neonatal and carrier screening, and assisted reproduction, and it represents a considerable advantage in CF genetic counseling.

The Italian External Quality Assessment Program for CF Sweat Chloride Test: Results of the 2015 Round Marco Salvatore, Giovanna Floridia, Annalisa Amato, Federica Censi, Claudio Carta, Maria Chiara de Stefano, Gianluca Ferrari, Fabrizio Tosto, Ettore Capoluongo, Ubaldo Caruso, Giuseppe Castaldo, Natalia Cirilli, Carlo Corbetta, Rita Padoan, Valeria Raia, Domenica Taruscio Abstract Background: Sweat chloride test is the gold standard test for cystic fibrosis (CF) diagnosis. In 2014 the Istituto Superiore di Sanità (ISS) established the first Italian pilot external quality assessment (EQA) program for CF sweat chloride test. In 2015 this activity was recognized as a third party service carried out by the ISS. The present paper describes the results of the first official 2015 sweat chloride test EQA program and results are compared with the 2014 round ones. Methods: the scheme is prospective; participation is open to Italian laboratories performing sweat test analysis for CF diagnosis. Enrollment is voluntary and since 2015 the payment of a fee is required. Participants are registered identified by an identification number known only to the ISS. Assessment covers analysis, interpretation and reporting. Results: thirteen laboratories, belonging to the Italian public Referral Centers for CF, participated in the 2015 round; nine already participated in 2014. Variability in scores of chloride titration and heterogeneity in interpretation / reporting results were identified in 2014 and 2015. Conclusions: results show variability in performance of laboratories indicating that the quality of laboratory performance is unpredictable unless EQA participation is mandatory as a component of compulsory laboratory accreditation.. Full Text: PDF DOI: 10.15640/jcb.v4n2a4