Begoña Laiz's research while affiliated with Hospital Universitari i Politècnic la Fe and other places

Hairy cell leukemia (HCL) is a relatively rare chronic B-cell malignancy that involves the bone marrow, spleen and peripheral blood. Monocytopenia could represent a clue for the suspicion of HCL with complete blood counts (CBCs) and careful assessment of the cell morphology being the first steps in the identification of hairy cells. The purpose of our study is to describe our experience with cell count and flag performance provided by the XN-module in a continuous series of six HCL patients diagnosed in the last two years. The final diagnosis was made by immunophenotypic and genetic analysis. Five out of six patients presented relative monocytosis on automated differential count. Nevertheless, the relative monocyte count was overestimated by the analyzer regarding to the manual count in all cases. The smear revision showed that most cells classified as monocytes were primarily hairy cells which afterwards were confirmed by the immunophenotype. All patients showed potentially pathologic WDF scatergrams or flags and were selected for microscopic smear review. In five of the six patients the WDF channel displayed the “Blasts/Abn Lympho?” flag and triggered the reflex reanalysis using the WPC channel. All samples presented lack or abnormal position of the monocytes cluster in the WPC scattergram. As a conclusion the monocytopenia should be taken with caution for the initial screening of hairy cell leukemia. Instead, an abnormal appearance of WDF scattergram and the lack or an ectopic position of the monocyte cluster in WPC scattergram should be considered to initiate the review of the peripheral blood smear.

Background To analyze the relationship of the antigen carbohydrate 125 (CA125) biomarker with the cellular rejection of the heart graft during the first year after transplantation. Methods Retrospective study of consecutive heart transplant (HTx) patients for 1.5 years. The total number of patients included in the study was 23 with a total of 103 follow-ups. In all patients, CA125 was determined before HTx and determined post-HTx in every follow-up. These were performed during months 1, 2, 4, 6, 9, and 12. Endomyocardial biopsy was performed in all revisions to assess the degree of graft rejection in the pathologic study. The biopsy results were grouped into 1. absence of rejection and 2. presence of some degree of rejection. Results The mean pretransplant CA125 value presented a median of 120 U/mL with an interquartile range of 28.8 U/mL. One month after transplantation, the value was reduced by 20% and at 2 months by 81%. In subsequent reviews, plasma values were always between 10 and 20 U/mL. When comparing the values by periods and according to the presence or absence of rejection, no significant differences were found other than a slight elevation at the 6-month checkup (P = .03) but without clinical relevance, because the CA125 value was slightly higher in biopsy results without rejection. Conclusion The rapid reduction of CA125 corroborates its usefulness as a marker of congestion in heart failure. This biomarker is not useful for predicting rejection. However, in cases of very severe rejections that occurred with systemic congestion, it could be raised. It would be necessary to corroborate this hypothesis in a larger study with a higher number of severe rejections.

Introducción: Alemtuzumab es un anticuerpo monoclonal anti-CD52 utilizado en el tratamiento de los pacientes con esclerosis multiple. Entre sus posibles efectos adversos destaca el riesgo de desarrollo de disfuncion tiroidea. El objetivo de este trabajo fue conocer la frecuencia de alteraciones tiroideas tras inicio del tratamiento e identificar factores de riesgo para la enfermedad tiroidea. Material y métodos: Se realizo un estudio retrospectivo, observacional, descriptivo de una serie de casos que incluyo a todos los pacientes con esclerosis multiple que recibieron tratamiento con alemtuzumab desde 2015 hasta 2019 en el Hospital Universitai i Politecnic La Fe, Valencia. Resultados: Se incluyeron en el estudio 43 pacientes. El 40 % de los pacientes (17 de 43) presentaron algun tipo de enfermedad tiroidea; 8 casos desarrollaron enfermedad de Graves, 2 de ellos con variante fluctuante, 3 casos tiroiditis Hashimoto, 2 casos hipotiroidismo, 2 casos hipotiroidismo subclinico transitorio, 1 caso hipertiroidismo sin anticuerpos antirreceptor de TSH (tirotropina) y 1 caso control dificil del tratamiento sustitutivo con levotiroxina tras la administracion del alemtuzumab, con una relacion tiempo dependiente entre el inicio del tratamiento y el desarrollo de la enfermedad tiroidea. El 75 % de los pacientes presentaron el primer episodio de disfuncion tiroidea dentro de los dos primeros años posadministracion de la primera dosis de alemtuzumab. Disfunción tiroidea inducida por alemtuzumab en pacientes con esclerosis multiple. Conclusión: Ante el alto riego de desarrollo de enfermedad tiroidea tras tratamiento con alentulemtuzumab se deberia realizar un cribado y seguimiento de la funcion tiroidea para un control eficaz de los posibles eventos adversos que puedan ocurrir en la evolucion de estos pacientes.

The establishment of precision medicine in cancer patients requires the study of several biomarkers. Single-gene testing approaches are limited by sample availability and turnaround time. Next generation sequencing (NGS) provides an alternative for detecting genetic alterations in several genes with low sample requirements. Here we show the implementation to routine diagnostics of a NGS assay under International Organization for Standardization (UNE-EN ISO 15189:2013) accreditation. For this purpose, 106 non-small cell lung cancer (NSCLC) and 102 metastatic colorectal cancer (mCRC) specimens were selected for NGS analysis with Oncomine Solid Tumor (ThermoFisher). In NSCLC the most prevalently mutated gene was TP53 (49%), followed by KRAS (31%) and EGFR (13%); in mCRC, TP53 (50%), KRAS (48%) and PIK3CA (16%) were the most frequently mutated genes. Moreover, NGS identified actionable genetic alterations in 58% of NSCLC patients, and 49% of mCRC patients did not harbor primary resistance mechanisms to anti-EGFR treatment. Validation with conventional approaches showed an overall agreement >90%. Turnaround time and cost analysis revealed that NGS implementation is feasible in the public healthcare context. Therefore, NGS is a multiplexed molecular diagnostic tool able to overcome the limitations of current molecular diagnosis in advanced cancer, allowing an improved and economically sustainable molecular profiling.

Background There is a lack of evidence about the loss of efficacy of anti-TNF agents in JIA and its possible relation to the immunogenicity generated. In the last years there has been interest in the role of serum calprotectin in children diseases, including JIA. Objectives To assess the immunogenicity and bioavailability of anti-TNF drugs, their relationship with disease activity and the clinical utility of serum calprotectin in monitoring JIA patients. Methods This is a 12-month prospective, multicenter, non-interventional, observational study. Patients from 2 to 18 years diagnosed with non-systemic JIA according to ILAR criteria and receiving treatment with IFX, ADA or ETN were included. The patients were evaluated using the CHAQ and the juvenile arthritis disease activity score (JADAS 71) and determination of anti-TNF drug, anti-drug antibody and calprotectina serum levels were performed. Results 222 patients were included. At month 12, 181 (81.5%) continued receiving treatment, and 205 (94.5%) had positive serum levels of anti-TNF drug (96.2% of ETN, 93.5% of ADA and 66.7% of IFX) at baseline and at 12 months serum levels were positive in 161 (95.3%) patients (97.5% ETN, 93.2% ADA and 100% IFX). In total, 16 (7.3%) patients presented anti-drug A (1 of 106 anti-ETN, 13 of 109 anti-ADA, 2 of 3 anti-IFX) at baseline and 4 patients (2.4%) (0 of 81 anti-ETN, 4 of 88 anti-ADA, 0 of 1 anti-IFX) at 12 months. Regarding the relationship between anti-TNF levels or anti-drug Abs, disease activity, and functional disability overall, statistically significant differences were not observed between the groups. Patients with high levels of serum calprotectin at the baseline visit (16 of 216) showed higher JADAS-71 score [1.23 (2.06 SD) vs. 2.06 (3.44 SD)] and CHAQ [0.11 SD (0.28 SD)] vs 0.17 (0.40 SD)] compared with normal calprotectin group, although no statistically significant differences were observed (p = 0.066 and p = 0.288) between the groups with a normal and high level of serum calprotectin and the number of patients was small. Conclusion The low prevalence of anti-drug Abs observed was in consonance with the high proportion of patients with a positive serum level of anti-TNF drug. These data suggest an appropriate management of the long-term treatment of Spanish JIA patients, who showed a maintained inactive disease/low disease activity state and a very low functional disability state as a consequence of a low immunogenicity and good bioavailability of anti-TNF drugs. View this table: • View inline • View popup • Download powerpoint Baseline patients’ characteristics. View this table: • View inline • View popup Table 2 JADAS -71 score according to immunogenicity and bioavailability of anti-TNF drugs Disclosure of Interests Inmaculada Calvo Grant/research support from: received research grants from Pfizer, Roche, Novartis, Clementia, Sanofi, MSD, BMS and GSK, Consultant for: Advisory boards: Novartis, AbbVie, Speakers bureau: AbbVie, Roche, Novartis, SOBI, Consuelo Modesto: None declared, Maria Montoro Shareholder of: Maria Montoro is employee of and shareholder in Pfizer, Employee of: Maria Montoro is employee of and shareholder in Pfizer, Begoña Laiz: None declared, Estibaliz Iglesias: None declared, Estefanía Quesada-Masachs: None declared, B Lopez-Montesinos: None declared, R Bou: None declared, M.I. Gonzalez-Fernández: None declared, J Fornés: None declared, A Rodriguez: None declared, jJ Sanchez: None declared, J Calzada: None declared, Tamara Rodriguez: None declared, Mireia Lopez Corbeto: None declared, Susana Gómez Employee of: I am a current employee of Pfizer., Jordi Anton Grant/research support from: JA has received grant/research support, consulting fees and/or honoraria from AbbVie, Alexion, BMS, ChemoCentryx, Gebro, GSK, Novartis, Novimmune, Pfizer, Roche, Sanofi and Sobi, Consultant for: JA has received grant/research support, consulting fees and/or honoraria from AbbVie, Alexion, BMS, ChemoCentryx, Gebro, GSK, Novartis, Novimmune, Pfizer, Roche, Sanofi and Sobi

Table S1 Ethics Committee approval details Table S2 Participant recruitment by study site Table S3 Comparison of angiogenic factors at 20, 24 and 28 weeks in women with singleton vs multiple pregnancy Table S4 Performance of different prediction models using soluble fms‐like tyrosine kinase‐1 (sFlt‐1)/placental growth factor (PlGF) ratio, sFlt‐1 or PlGF to estimate risk of early‐onset pre‐eclampsia (PE) at 20, 24 and 28 weeks Figure S1 Formulae used in prediction model for early‐onset pre‐eclampsia (PE). MAP, mean arterial pressure. Figure S2 Nomogram for estimation of risk of early‐onset pre‐eclampsia (PE) at 24 weeks. To calculate probability of early‐onset PE for a given patient, the value for each predictor is obtained by drawing a vertical line straight upward from that factor to the ‘points’ axis. The points are then summed and the sum located on the total points nomogram, and the probability of early‐onset PE is located by drawing a vertical line downward to the ‘risk of PE’ line. MAP, mean arterial pressure; PlGF, placental growth factor; sFlt‐1, soluble fms‐like tyrosine kinase‐1. Figure S3 Worked example of the use of the nomogram for estimation of risk of early‐onset pre‐eclampsia (PE) at 24 weeks. To calculate visually risk of early‐onset PE in a parous patient with previous PE at 24 weeks, with mean arterial pressure (MAP) of 100 mmHg and soluble fms‐like tyrosine kinas‐1 (sFlt‐1)/placental growth factor (PlGF) ratio of 50, points are assigned for each item by plotting a line from the item to the points line. Being parous equates to 0 points; 100 mmHg MAP corresponds to 9 points; previous PE equates to 38 points; and sFlt‐1/PlGF ratio of 50 (In = 3.91) gives 62 points. Total number of points is 109; drawing a vertical line downward from total points axis to ‘risk of PE’ line, the risk is estimated at approximately 80%. Figure S4 Placental growth factor (PlGF) (a) and soluble fms‐like tyrosine kinase‐1 (sFlt‐1) (b) at 20, 24 and 28 weeks in women who developed early‐onset pre‐eclampsia (PE) and in control participants who did not develop PE.

Objectives: Accurate evaluation of analyzers is highly recommended before these devices are broadly introduced for routine testing. Concerning quantification of IgG subclasses (IgGSc), standardization has not yet been reached and thus different assays might lead to different results. Here we report the analytical performances of The Binding Site (TBS) SPAPLUS® human IgGSc assay and the concordance with the Siemens BNII® human IgGSc assay. Design and methods: We evaluated precision, LoB, LoD and linearity of TBS SPAPLUS® human IgGSc immunoassay. Quantitation of IgGSc in 53 patients' serum samples was performed in parallel on both analyzers. Results from both assays were compared. Results: Analytical performances of the TBS SPAPLUS® human IgGSc assay are acceptable for routine clinical use. According to the method comparison study, TBS assay measures lower values than Siemens assay for IgG1 and IgG4, whereas for IgG2 and IgG3 TBS provides greater values. All assays present a proportional bias, greater in the case of IgG3 and IgG4 assays. Individual subclass agreement, based on the classification of samples within three categories (low, normal and high) according to assay-specific reference intervals, range from 75% (IgG1) to 92% (IgG2). However, total classification agreement over all four subclasses only account for 55% of samples. Conclusion: Results obtained from both assays are not interchangeable. Standardization of IgGSc assay and review of the reference ranges must be accomplished in order to achieve a higher degree of agreement between different methods.

Background: Accurate evaluation of hematology analyzers is recommended before these devices can be broadly introduced for the routine testing of continuous ambulatory peritoneal dialysis (CAPD), ascitic, and pleural fluids. Methods: We evaluated the performance of Mindray BC-6800 for white blood cell (WBC) and differential cell count in 50 CAPD, 60 ascitic and 40 pleural compared with manual microscopy. Within-run precision, limit of blank (LoB), limit of detection (LoD), limit of quantitation (LoQ), and carryover were assessed. Results: The Passing-Bablok regression in all fluids showed the following equations: yWBC =1.05x+3.31 (95%CI slope 0.95 to 1.12; intercept -0.25 to 5.52); yMN =0.85x+15.63 (95%CI slope 0.72 to 1.05; intercept -24.18 to 84.47); and yPMN =1.21x+13.37 (95%CI slope 1.03 to 1.35; intercept 4.00 to 32.47) with bias 78 cells/μL. The AUC for clinical PMN cut-off was 0.88 (95%CI: 0.77 to 0.98). In ascitic, pleural, and CAPD fluids the AUC for clinical PMN cut-off were 0.88 (95%CI: 0.63 to 1.00), 0.83 (95%CI: 0.68 to 0.99), and 1.00 (95%CI: 1.00 to 1.00) respectively. CV ranged from 3%-34%. LoB of 3 cell/μL was verified. LoD and LoQ reported the same result (8 cells/μL). Carry over never exceeded 0.05%. Conclusion: The effectiveness of BC-6800 to categorize cells from different body fluids was not compromised by the slight positive bias observed. This conclusion is supported by the high AUC and agreement between the automated method and the reference method. The results show that BC-6800 offers rapid, accurate, and reproducible results for clinical management of CAPD, ascitic, and pleural fluids.