Using Microfluidic Hepatic Spheroid Cultures to Assess Liver Toxicity of T-2 Mycotoxin.

The Fusarium fungi is found in cereals and feedstuffs and may produce mycotoxins, which are secondary metabolites, such as the T-2 toxin (T-2). In this work, we explored the hepatotoxicity of T-2 using microfluidic 3D hepatic cultures. The objectives were: (i) exploring the benefits of microfluidic 3D cultures compared to conventional 3D cultures available commercially (Aggrewell plates), (ii) establishing 3D co-cultures of hepatic cells (HepG2) and stellate cells (LX2) and assessing T-2 exposure in this model, (iii) characterizing the induction of metabolizing enzymes, and (iv) evaluating inflammatory markers upon T-2 exposure in microfluidic hepatic cultures. Our results demonstrated that, in comparison to commercial (large-volume) 3D cultures, spheroids formed faster and were more functional in microfluidic devices. The viability and hepatic function decreased with increasing T-2 concentrations in both monoculture and co-cultures. The RT-PCR analysis revealed that exposure to T-2 upregulates the expression of multiple Phase I and Phase II hepatic enzymes. In addition, several pro- and anti-inflammatory proteins were increased in co-cultures after exposure to T-2.

Low-Cost EEG Multi-Subject Recording Platform for the Assessment of Students' Attention and the Estimation of Academic Performance in Secondary School.

The level of student attention in class greatly affects their academic performance. Teachers typically rely on visual inspection to react to students' attention in time, but this subjective method leads to inconsistencies across classes. Online education exacerbates the issue as students can turn off cameras and microphones to keep their own privacy. To address this, we present a novel, low-cost EEG-based platform for assessing students' attention and estimating their academic performance. In a study involving 34 secondary school students (aged 14 to 16), participants watched an academic video and answered evaluation questions while their EEG activity was recorded using a commercial headset. The results demonstrate a significant correlation (0.53, p-value = 0.003) between the power spectral density (PSD) of the EEG beta band (12-30 Hz) and students' academic performance. Additionally, there was a notable difference in PSD-beta between high and low academic performers. These findings encourage the use of PSD-beta for the immediate and objective assessment of both the student attention and the subsequent academic performance. The platform offers valuable and objective feedback to teachers, enhancing the effectiveness of both face-to-face and online teaching and learning environments.

Design and Validation of a Custom Next-Generation Sequencing Panel in Pediatric Acute Lymphoblastic Leukemia.

The molecular landscape of acute lymphoblastic leukemia (ALL) is highly heterogeneous, and genetic lesions are clinically relevant for diagnosis, risk stratification, and treatment guidance. Next-generation sequencing (NGS) has become an essential tool for clinical laboratories, where disease-targeted panels are able to capture the most relevant alterations in a cost-effective and fast way. However, comprehensive ALL panels assessing all relevant alterations are scarce. Here, we design and validate an NGS panel including single-nucleotide variants (SNVs), insertion-deletions (indels), copy number variations (CNVs), fusions, and gene expression (ALLseq). ALLseq sequencing metrics were acceptable for clinical use and showed 100% sensitivity and specificity for virtually all types of alterations. The limit of detection was established at a 2% variant allele frequency for SNVs and indels, and at a 0.5 copy number ratio for CNVs. Overall, ALLseq is able to provide clinically relevant information to more than 83% of pediatric patients, making it an attractive tool for the molecular characterization of ALL in clinical settings.

Impact of pre- and/or post-autologous stem cell transplantation exposure to brentuximab vedotin on survival outcomes in patients with high-risk Hodgkin lymphoma.

The AETHERA trial demonstrated that brentuximab vedotin (BV) consolidation after autologous stem cell transplantation (ASCT) in patients with Hodgkin lymphoma (HL) at high risk of relapse/progression increases progression-free survival (PFS). Patients previously exposed to BV were excluded from that trial. However, BV alone or in combination with chemotherapy is frequently used as front-line treatment and/or pre-ASCT salvage therapy. We analyzed data from 156 patients with high-risk HL who underwent ASCT with (BV-CON, n = 62) or without (non-BV, n = 94) BV consolidation. Fifty-seven patients received BV-based salvage regimens before ASCT. The 3-year overall survival and PFS for all patients were 91.6% and 70.0%, respectively. Multivariate analysis showed that BV-CON was associated with better PFS (HR 0.39, p = 0.01), whereas positive PET at transplant leaded to worse PFS (HR 2.71, p = 0.001). BV-CON improved PFS in PET-positive patients (72.2% vs. 43.0%, p = 0.05), with a beneficial trend observed in PET negative (88.8% vs. 75.2%, p = 0.09). BV-CON patients with or without BV exposure pre-ASCT had a significantly better PFS than non-BV with or without BV pretransplant treatment (HR 0.36, p = 0.004). The efficacy of real-life BV consolidation therapy was similar to that in the AETHERA trial. This therapeutic strategy improves survival independently of BV exposure prior to ASCT.

[Validity and reliability of the 8-item EUROHIS-QOL to assess Brazilian adults' quality of life]. / Validade e confiabilidade do EUROHIS-QOL 8-item para avaliar a qualidade de vida em adultos brasileiros.

The study evaluated the psychometric properties of the EUROHIS-QOL 8-item scale in Brazilian adults. The EUROHIS-QOL is a measurement of quality of life developed based on the generic instruments of WHOQOL-100 and WHOQOL-BREF. Based on data from a prospective cohort of 1,100 adults, participants in the baseline in 2006 and 2007, exploratory factor analysis, internal consistency, reliability, and discriminant validity were performed via multitrait-multimethod analysis, with the calculation of the average variance extracted (AVE). Confirmatory factor analysis was conducted with 573 of these participants in 2013 to 2018. Solutions were tested with one and with two factors named functional capacity and socioeconomic conditions. Both solutions presented good internal consistency and reliability. The correlation between the items was 0.535, and the AVE was 0.397 for solution with one factor, and 0.528 and 0.341 for factors one and two, respectively, indicating good discriminant validity in the bifactorial solution. Both factors had eigenvalues greater than one and factor loadings ranging from 0.398 to 0.915. The adjustment of the unifactorial solution presented: χ2 = 186 (g.l. = 18), p < 0.001, RMSEA = 0.128 (90%CI: 0.111-0.145), CFI = 0.960, TLI = 0.938, and SMRM = 0.042; while in the bifactorial presented: χ2 = 135 (g.l. = 17), p < 0.001, RMSEA = 0.110 (90%CI: 0.093-0.128), CFI = 0.972, TLI = 0.954, and SMRM = 0.035. Overall, the results provide reasonable evidence for the construct validity and reliability of the EUROHIS-QOL 8-item. Instruments with fewer items, as in our case, provide greater practicality and are ideal for use in population-based studies with extensive samples.

O estudo avaliou as propriedades psicométricas da escala EUROHIS-QOL 8-item em adultos brasileiros. O EUROHIS-QOL 8-item é uma medida de qualidade de vida desenvolvida a partir dos instrumentos genéricos WHOQOL-100 e WHOQOL-BREF. A partir dos dados de uma coorte prospectiva com 1.100 adultos participantes da linha de base em 2006 e 2007, foram realizadas análise fatorial exploratória, consistência interna, confiabilidade e validade discriminante por meio da análise multitraço-multimétodo, com o cálculo das variâncias médias extraídas (AVE). A análise fatorial confirmatória foi conduzida com 573 desses participantes nos anos de 2013 a 2018. Foram testadas soluções com um e com dois fatores nomeados capacidade funcional e condições socioeconômicas. Ambas as soluções apresentaram boa consistência interna e confiabilidade. A correlação entre os itens foi de 0,535 e as AVE foram 0,397 para solução com um fator, e 0,528 e 0,341 para os fatores um e dois, respectivamente, indicando boa validade discriminante na solução bifatorial. Os dois fatores tiveram autovalores maiores que um e cargas fatoriais variando de 0,398 a 0,915. O ajustamento da solução unifatorial foi: χ2 = 186 (g.l. = 18), p < 0,001, RMSEA = 0,128 (IC90%: 0,111-0,145), CFI = 0,960, TLI = 0,938 e SMRM = 0,042; enquanto na bifatorial era: χ2 = 135 (g.l. = 17), p < 0,001, RMSEA = 0,110 (IC90%: 0,093-0,128), CFI = 0,972, TLI = 0,954 e SMRM = 0,035. De modo geral, os resultados fornecem evidências razoáveis da validade de construto e confiabilidade do EUROHIS-QOL 8-item. Instrumentos com menor número de itens, como é o caso, proporcionam maior praticidade e são ideais para utilização em estudos de base populacional com uso de amostras extensas.

El estudio evaluó las propiedades psicométricas de la escala EUROHIS-QOL 8-ítems (Europe Health Interview Surveys Quality of Life Abbreviated Instrument) en adultos brasileños. El índice de 8 ítems EUROHIS-QOL es una medida de calidad de vida desarrollada con base en los instrumentos genéricos WHOQOL-100 (World Health Organization Quality of Life Instrument) y WHOQOL-BREF (World Health Organization Quality of Life Abbreviated Instrument). Con base en los datos de una cohorte prospectiva de 1.100 adultos que participaron en la línea de base en 2006 y 2007, se realizaron los análisis factorial exploratorio, de consistencia interna, de confiabilidad y de validez discriminante mediante un análisis multirrasgo-multimétodo, con el cálculo de la varianza media extraída (AVE). Se realizó un análisis factorial confirmatorio con 573 de estos participantes de 2013 hasta 2018. Se probaron soluciones de uno y dos factores denominados capacidad funcional y condiciones socioeconómicas. Ambas soluciones presentaron buena consistencia interna y confiabilidad. La correlación entre los ítems fue de 0,535 y las AVE fueron de 0,397 para la solución de un factor y de 0,528 y 0,341 para los factores uno y dos, respectivamente, lo que indica una buena validez discriminante en la solución bifactorial. Los dos factores tuvieron autovalores mayores a uno y cargas factoriales que oscilaron entre 0,398 y 0,915. El ajuste de la solución unifactorial fue: χ2 = 186 (g.l. = 18), p < 0,001, RMSEA = 0,128 (IC90%: 0,111-0,145), CFI = 0,960, TLI = 0,938 y SMRM = 0,042; mientras que en la bifactorial fue: χ2 = 135 (g.l. = 17), p < 0,001, RMSEA = 0,110 (IC90%: 0,093-0,128), CFI = 0,972, TLI = 0,954 e SMRM = 0,035. En general, los resultados proporcionan evidencias razonables de la validez de constructo y confiabilidad del EUROHIS-QOL 8-ítems. Los instrumentos con menor número de ítems, como es el caso, proporcionan mayor practicidad y son ideales para su uso en estudios de base poblacional con muestras amplias.

Panel Sequencing for Clinically Oriented Variant Screening and Copy Number Detection in Chronic Lymphocytic Leukemia Patients.

According to current guidelines, in chronic lymphocytic leukemia (CLL), only the TP53 molecular status must be evaluated prior to every treatment's initiation. However, additional heterogeneous genetic events are known to confer a proliferative advantage to the tumor clone and are associated with progression and treatment failure in CLL patients. Here, we describe the implementation of a comprehensive targeted sequencing solution that is suitable for routine clinical practice and allows for the detection of the most common somatic single-nucleotide and copy number variants in genes relevant to CLL. We demonstrate that this cost-effective strategy achieves variant detection with high accuracy, specificity, and sensitivity. Furthermore, we identify somatic variants and copy number variations in genes with prognostic and/or predictive value, according to the most recent literature, and the tool provides evidence about subclonal events. This next-generation sequencing (NGS) capture-based target assay is an improvement on current approaches in defining molecular prognostic and/or predictive variables in CLL patients.

Validade e confiabilidade do EUROHIS-QOL 8-item para avaliar a qualidade de vida em adultos brasileiros / Validity and reliability of the 8-item EUROHIS-QOL to assess Brazilian adults' quality of life / Validez y confiabilidad de EUROHIS-QOL-8-ítem para evaluar la calidad de vida en adultos brasileños

O estudo avaliou as propriedades psicométricas da escala EUROHIS-QOL 8-item em adultos brasileiros. O EUROHIS-QOL 8-item é uma medida de qualidade de vida desenvolvida a partir dos instrumentos genéricos WHOQOL-100 e WHOQOL-BREF. A partir dos dados de uma coorte prospectiva com 1.100 adultos participantes da linha de base em 2006 e 2007, foram realizadas análise fatorial exploratória, consistência interna, confiabilidade e validade discriminante por meio da análise multitraço-multimétodo, com o cálculo das variâncias médias extraídas (AVE). A análise fatorial confirmatória foi conduzida com 573 desses participantes nos anos de 2013 a 2018. Foram testadas soluções com um e com dois fatores nomeados capacidade funcional e condições socioeconômicas. Ambas as soluções apresentaram boa consistência interna e confiabilidade. A correlação entre os itens foi de 0,535 e as AVE foram 0,397 para solução com um fator, e 0,528 e 0,341 para os fatores um e dois, respectivamente, indicando boa validade discriminante na solução bifatorial. Os dois fatores tiveram autovalores maiores que um e cargas fatoriais variando de 0,398 a 0,915. O ajustamento da solução unifatorial foi: χ2 = 186 (g.l. = 18), p < 0,001, RMSEA = 0,128 (IC90%: 0,111-0,145), CFI = 0,960, TLI = 0,938 e SMRM = 0,042; enquanto na bifatorial era: χ2 = 135 (g.l. = 17), p < 0,001, RMSEA = 0,110 (IC90%: 0,093-0,128), CFI = 0,972, TLI = 0,954 e SMRM = 0,035. De modo geral, os resultados fornecem evidências razoáveis da validade de construto e confiabilidade do EUROHIS-QOL 8-item. Instrumentos com menor número de itens, como é o caso, proporcionam maior praticidade e são ideais para utilização em estudos de base populacional com uso de amostras extensas.

The study evaluated the psychometric properties of the EUROHIS-QOL 8-item scale in Brazilian adults. The EUROHIS-QOL is a measurement of quality of life developed based on the generic instruments of WHOQOL-100 and WHOQOL-BREF. Based on data from a prospective cohort of 1,100 adults, participants in the baseline in 2006 and 2007, exploratory factor analysis, internal consistency, reliability, and discriminant validity were performed via multitrait-multimethod analysis, with the calculation of the average variance extracted (AVE). Confirmatory factor analysis was conducted with 573 of these participants in 2013 to 2018. Solutions were tested with one and with two factors named functional capacity and socioeconomic conditions. Both solutions presented good internal consistency and reliability. The correlation between the items was 0.535, and the AVE was 0.397 for solution with one factor, and 0.528 and 0.341 for factors one and two, respectively, indicating good discriminant validity in the bifactorial solution. Both factors had eigenvalues greater than one and factor loadings ranging from 0.398 to 0.915. The adjustment of the unifactorial solution presented: χ2 = 186 (g.l. = 18), p < 0.001, RMSEA = 0.128 (90%CI: 0.111-0.145), CFI = 0.960, TLI = 0.938, and SMRM = 0.042; while in the bifactorial presented: χ2 = 135 (g.l. = 17), p < 0.001, RMSEA = 0.110 (90%CI: 0.093-0.128), CFI = 0.972, TLI = 0.954, and SMRM = 0.035. Overall, the results provide reasonable evidence for the construct validity and reliability of the EUROHIS-QOL 8-item. Instruments with fewer items, as in our case, provide greater practicality and are ideal for use in population-based studies with extensive samples.

El estudio evaluó las propiedades psicométricas de la escala EUROHIS-QOL 8-ítems (Europe Health Interview Surveys Quality of Life Abbreviated Instrument) en adultos brasileños. El índice de 8 ítems EUROHIS-QOL es una medida de calidad de vida desarrollada con base en los instrumentos genéricos WHOQOL-100 (World Health Organization Quality of Life Instrument) y WHOQOL-BREF (World Health Organization Quality of Life Abbreviated Instrument). Con base en los datos de una cohorte prospectiva de 1.100 adultos que participaron en la línea de base en 2006 y 2007, se realizaron los análisis factorial exploratorio, de consistencia interna, de confiabilidad y de validez discriminante mediante un análisis multirrasgo-multimétodo, con el cálculo de la varianza media extraída (AVE). Se realizó un análisis factorial confirmatorio con 573 de estos participantes de 2013 hasta 2018. Se probaron soluciones de uno y dos factores denominados capacidad funcional y condiciones socioeconómicas. Ambas soluciones presentaron buena consistencia interna y confiabilidad. La correlación entre los ítems fue de 0,535 y las AVE fueron de 0,397 para la solución de un factor y de 0,528 y 0,341 para los factores uno y dos, respectivamente, lo que indica una buena validez discriminante en la solución bifactorial. Los dos factores tuvieron autovalores mayores a uno y cargas factoriales que oscilaron entre 0,398 y 0,915. El ajuste de la solución unifactorial fue: χ2 = 186 (g.l. = 18), p < 0,001, RMSEA = 0,128 (IC90%: 0,111-0,145), CFI = 0,960, TLI = 0,938 y SMRM = 0,042; mientras que en la bifactorial fue: χ2 = 135 (g.l. = 17), p < 0,001, RMSEA = 0,110 (IC90%: 0,093-0,128), CFI = 0,972, TLI = 0,954 e SMRM = 0,035. En general, los resultados proporcionan evidencias razonables de la validez de constructo y confiabilidad del EUROHIS-QOL 8-ítems. Los instrumentos con menor número de ítems, como es el caso, proporcionan mayor practicidad y son ideales para su uso en estudios de base poblacional con muestras amplias.

Severe infections in patients with lymphoproliferative diseases treated with new targeted drugs: A multicentric real-world study.

BACKGROUND: Lymphoid neoplasms treatment has recently been renewed to increase antitumor efficacy and conventional chemotherapies toxicities. Limited data have been published about the infection risk associated with these new drugs, therefore this study analyzes the infectious complications in patients with lymphoproliferative diseases (LPD) treated with monoclonal antibodies (obinutuzumab, ofatumumab, brentuximab, nivolumab, or pembrolizumab), BTK inhibitors (ibrutinib and acalabrutinib), PI3K inhibitors (idelalisib) and BCL2 inhibitors (venetoclax). METHODS: Multicenter retrospective study of 458 LPD patients treated with targeted therapies in real-life setting, in 18 Spanish institutions, from the time of their commercial availability to August 2020. RESULTS: Severe infections incidence was 23% during 17-month median follow-up; cumulative incidence was higher in the first 3-6 months of targeted drug treatment and then decreased. The most frequent etiology was bacterial (54%). Nine (6%) Invasive fungal infections (IFI) were observed, in its majority in chronic lymphocytic leukemia (CLL) patients treated predominantly with ibrutinib. Significant risk factors for severe infection were: severe lymphopenia (p = 0.009, OR 4.7, range 1.3-1.7), combined targeted treatment vs single agent treatment (p = 0.014 OR 2.2 range 1.1-4.2) and previous rituximab (p = 0.03 OR 1.8, range 1.05-3.3). Infection-related mortality was 6%. In 22% of patients with severe infections, definitive discontinuation of the targeted drug was observed. CONCLUSION: A high proportion of patients presented severe infections during follow-up, with non-negligible attributable mortality, but infection incidence is not superior to the one observed during the chemotherapy era. In selected cases with specific risk factors for infection, antimicrobial prophylaxis should be considered.

Addition of chemotherapy to nivolumab after PD-1 inhibitor failure as bridge to allogeneic stem cell transplantation in classical Hodgkin's lymphoma: report on three cases and literature review.

The poor prognosis of refractory or relapsed (R/R) classical Hodgkin's lymphoma (cHL) after autologous stem cell transplantation has improved greatly due to the introduction of brentuximab vedotin and PD-1 inhibitors. However, the duration of response achieved with these novel agents is too short. The information about the management of patients after anti-PD-1 therapy failure is very limited in cHL, although chemotherapy alone or combined with PD-1 inhibitors has shown encouraging results. We report three cases of heavily pretreated cHL, refractory to nivolumab monotherapy, successfully rescued with the addition of chemotherapy to nivolumab, as a bridge to allogeneic stem cell transplantation (allo-SCT). All three patients presented poor clinical features such as three to four previous lines including brentuximab vedotin and autologous stem cell transplantation, refractoriness to the last line of therapy previous to nivolumab, and rapid disease progression. Notwithstanding these characteristics and nivolumab failure, they achieved a complete response after the addition of chemotherapy, were consolidated with allo-SCT, and still remain in complete response. There are few studies concerning the combination of PD-1 inhibitors and chemotherapy after nivolumab failure, including one retrospective study and one phase II trial with nivolumab plus bendamustine. Therefore, only few patients are consolidated with allo-SCT. However, there are several ongoing trials investigating new combinations of chemotherapy and PD-1 inhibitors in R/R cHL, as well as in first line. All these data suggest that anti-PD-1 therapy may reprogram the immune system, activating and inhibiting effector and immunosuppressive cells, respectively, leading to overtake of chemorefractoriness. Allo-SCT can increase the immune-related events of patients treated with anti-PD-1 previously, consistent on acute graft-versus-host disease, sinusoidal obstruction syndrome, and noninfectious febrile syndrome. In conclusion, the combination of PD-1 inhibitor and chemotherapy may be a feasible therapy after anti-PD-1 treatment failure as a bridge to allo-SCT.

Central Nervous System Involvement in Epstein-Barr Virus-Related Post-Transplant Lymphoproliferative Disorders after Allogeneic Hematopoietic Stem Cell Transplantation.

Central nervous system (CNS) involvement in Epstein-Barr virus-related post-transplant lymphoproliferative disorders (EBV-PTLDs) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) is poorly defined. We analyzed the incidence, clinical and pathological characteristics, and impact on outcomes of EBV-PTLDs with CNS involvement (CNS-PTLDs) in 1009 consecutive adult patients undergoing allo-HSCT at a single-center institution. Four hundred eighty-two patients received matched sibling donor (MSD) transplants, 388 umbilical cord blood transplants (UCBTs), 56 matched unrelated donor (MUD) transplants, and 83 haploidentical transplants. We detected 25 cases of biopsy-proven EBV-PTLDs. Of these, nine patients (36%) had CNS-PTLDs: six after UCBT (67%), one after MSD transplantation (11%), one after MUD transplantation (11%), and one after haploidentical transplantation (11%). The 5-year cumulative incidence risk of CNS-PTLDs was 0.9%. Median time from transplant to CNS-PTLDs was 187 days, and all patients had neurological symptoms at diagnosis. Six out of the nine cases (67%) occurred with systemic involvement, and three cases (33%) had isolated CNS involvement. The most frequent histological subtype was monomorphic EBV-PTLD, and laboratory characteristics were similar to EBV-PTLDs without CNS involvement. We observed statistical differences in the rate of positive EBV DNA detection in plasma between isolated CNS-PTLDs (detection in one out of three, 33%) and the rest of the EBV-PTLDs (100%) (P = .01). Treatment strategies included chemotherapy, radiotherapy, and T cell therapy. However, seven out of nine patients died due to progression of the CNS-PTLDs at a median time of 17 days (range, 8 to 163) from diagnosis. The 5-years overall survival in patients who developed CNS-PTLDs was 22% (95% confidence interval [CI], 7% to 75%) and 5-year treatment-related mortality was 78% (95% CI, 51% to 100%), with no statistically significant differences between CNS-PTLDs and the rest of the EBV-PTLDs. In conclusion, despite advances in EBV monitoring and treatment strategies, CNS-PTLDs remain an uncommon but serious complication after allo-HSCT, with very poor prognosis.