Flow cytometry immunophenotyping for diagnostic orientation and classification of pediatric cancer based on the euroflow solid tumor orientation tube (Stot)

© 2021 by the authors.Early diagnosis of pediatric cancer is key for adequate patient management and improved outcome. Although multiparameter flow cytometry (MFC) has proven of great utility in the diagnosis and classification of hematologic malignancies, its application to non-hematopoietic pediatric tumors remains limited. Here we designed and prospectively validated a new single eight-color antibody combination—solid tumor orientation tube, STOT—for diagnostic screening of pediatric cancer by MFC. A total of 476 samples (139 tumor mass, 138 bone marrow, 86 lymph node, 58 peripheral blood, and 55 other body fluid samples) from 296 patients with diagnostic suspicion of pediatric cancer were analyzed by MFC vs. conventional diagnostic procedures. STOT was designed after several design–test–evaluate–redesign cycles based on a large panel of monoclonal antibody combinations tested on 301 samples. In its final version, STOT consists of a single 8-color/12-marker antibody combination (CD99-CD8/numyogenin/CD4-EpCAM/CD56/GD2/smCD3-CD19/cyCD3-CD271/CD45). Prospective validation of STOT in 149 samples showed concordant results with the patient WHO/ICCC-3 diagnosis in 138/149 cases (92.6%). These included: 63/63 (100%) reactive/disease-free samples, 43/44 (98%) malignant and 4/4 (100%) benign non-hematopoietic tumors together with 28/38 (74%) leukemia/lymphoma cases; the only exception was Hodgkin lymphoma that required additional markers to be stained. In addition, STOT allowed accurate discrimination among the four most common subtypes of malignant CD45− CD56++ non-hematopoietic solid tumors: 13/13 (GD2++ numyogenin− CD271−/+ nuMyoD1− CD99− EpCAM−) neuroblastoma samples, 5/5 (GD2− numyogenin++ CD271++ nuMyoD1++ CD99−/+ EpCAM−) rhabdomyosarcomas, 2/2 (GD2−/+ numyogenin− CD271+ nuMyoD1− CD99+ EpCAM−) Ewing sarcoma family of tumors, and 7/7 (GD2− numyogenin− CD271+ nuMyoD1− CD99− EpCAM+) Wilms tumors. In summary, here we designed and validated a new standardized antibody combination and MFC assay for diagnostic screening of pediatric solid tumors that might contribute to fast and accurate diagnostic orientation and classification of pediatric cancer in routine clinical practice.This research was funded by the EuroFlow Consortium; Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro, Brazil (FAPERJ), numbers: E26/110.105/2014, E-26/010.101259/2018, and E26/102.191/2013; grant from Conselho Nacional de Desenvolvimento Científico e Tecnológico, Brasília, Brazil (CNPQ), Brasília, Brazil, numbers: 303765/2018-6, 409440/2016-7, and 400194/2014-7; and Instituto Desiderata/Chevron, Rio de Janeiro, Brazil, grant “Actions to improve pediatric cancer assistance in RJ”; the EuroFlow Consortium (grant LSHB-CT-2006-018708); Centro de Investigación Biomédica en Red de Cáncer (CIBER-ONC; Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, Madrid, Spain and FONDOS FEDER), numbers: CB16/12/00400, CB16/12/00233, CB16/12/00369, CB16/12/00489 and CB16/12/00480; grant from Bilateral Cooperation Program between Coordenação de Aperfeiçoamento de Pessoal de Nível Superior-CAPES (Brasília/Brazil) and Dirección General de Políticas Universitárias (DGPU)-Ministério de Educación, Cultura y Deportes (Madrid/Spain) number DGPU 311/15

Affective auditory stimuli: Adaptation of the International Affective Digitized Sounds (IADS-2) for European Portuguese

In this study, we present the normative values of the adaptation of the International Affective Digitized Sounds (IADS-2; Bradley & Lang, 2007a) for European Portuguese (EP). The IADS-2 is a standardized database of 167 naturally occurring sounds that is widely used in the study of emotions. The sounds were rated by 300 college students who were native speakers of EP, in the three affective dimensions of valence, arousal, and dominance, by using the Self-Assessment Manikin (SAM). The aims of this adaptation were threefold: (1) to provide researchers with standardized and normatively rated affective sounds to be used with an EP population; (2) to investigate sex and cultural differences in the ratings of affective dimensions of auditory stimuli between EP and the American (Bradley & Lang, 2007a) and Spanish (Fernandez-Abascal et al., Psicothema 20: 104-113 2008; Redondo, Fraga, Padron, & Pineiro, Behavior Research Methods 40: 784790 2008) standardizations; and (3) to promote research on auditory affective processing in Portugal. Our results indicated that the IADS-2 is a valid and useful database of digitized sounds for the study of emotions in a Portuguese context, allowing for comparisons of its results with those of other international studies that have used the same database for stimulus selection. The normative values of the EP adaptation of the IADS-2 database can be downloaded along with the online version of this article.This work is part of the research project “Procura Palavras (P-Pal): A Software Program for Deriving Objective and Subjective Psycholinguistic Indices for European Portuguese Words” (PTDC/PSI-PCO/104679/2008), funded by FCT (Fundação para a Ciência e Tecnologia), and FEDER (Fundo Europeu de Desenvolvimento Regional), through the European programs QREN (Quadro de Referência Estratégico Nacional), and COMPETE (Programa Operacional Factores de Competitividade)