Cytogenetic Assessment and Risk Stratification in Myelofibrosis with Optical Genome Mapping.

Cytogenetic assessment in myelofibrosis is essential for risk stratification and patient management. However, an informative karyotype is unavailable in a significant proportion of patients. Optical genome mapping (OGM) is a promising technique that allows for a high-resolution assessment of chromosomal aberrations (structural variants, copy number variants, and loss of heterozygosity) in a single workflow. In this study, peripheral blood samples from a series of 21 myelofibrosis patients were analyzed via OGM. We assessed the clinical impact of the application of OGM for disease risk stratification using the DIPSS-plus, GIPSS, and MIPSS70+v2 prognostic scores compared with the standard-of-care approach. OGM, in combination with NGS, allowed for risk classification in all cases, compared to only 52% when conventional techniques were used. Cases with unsuccessful karyotypes (n = 10) using conventional techniques were fully characterized using OGM. In total, 19 additional cryptic aberrations were identified in 9 out of 21 patients (43%). No alterations were found via OGM in 4/21 patients with previously normal karyotypes. OGM upgraded the risk category for three patients with available karyotypes. This is the first study using OGM in myelofibrosis. Our data support that OGM is a valuable tool that can greatly contribute to improve disease risk stratification in myelofibrosis patients.

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.

Microencapsulation Improved Fumaric Acid and Thymol Effects on Broiler Chickens Challenged With a Short-Term Fasting Period.

The first objective of this study was to demonstrate the usefulness of the microencapsulation technique to protect fumaric acid and thymol, avoiding their early absorption and ensuring their slow release throughout the gastrointestinal tract (GIT). For this purpose, the release of a lipid matrix microencapsulated brilliant blue (BB) was assessed in vitro, using a simulated broiler intestinal fluid, and in vivo. In vitro results showed that more than 60% of BB color reached the lower intestine, including 26.6 and 29.7% in the jejunum and ileum, respectively. The second objective was to determine the effects of microencapsulated fumaric acid, thymol, and their mixture on the performance and gut health of broilers challenged with a short-term fasting period (FP). One-day-old male ROSS 308 chickens (n = 280) were randomly distributed into seven treatments, with 10 replicates of four birds each. Dietary treatments consisted of a basal diet as negative control (NC), which was then supplemented by either non-microencapsulated fumaric acid (0.9 g/kg), thymol (0.6 g/kg), or a mixture of them. The same additive doses were also administered in a microencapsulated form (1.5 and 3 g/kg for the fumaric acid and thymol, respectively). At day 21, chickens were subjected to a 16.5-h short-term FP to induce an increase in intestinal permeability. Growth performance was assessed weekly. At day 35, ileal tissue and cecal content were collected from one bird per replicate to analyze intestinal histomorphology and microbiota, respectively. No treatment effect was observed on growth performance from day 1 to 21 (p > 0.05). Microencapsulated fumaric acid, thymol, or their mixture improved the overall FCR (feed conversion ratio) and increased ileal villi height-to-crypt depth ratio (VH:CD) (p < 0.001) on day 35 of the experiment. The microencapsulated mixture of fumaric acid and thymol increased cecal abundance of Bacteroidetes, Bacillaceae, and Rikenellaceae, while decreasing that of Pseudomonadaceae. These results indicate that the microencapsulation technique used in the current study can be useful to protect fumaric acid and thymol, avoiding early absorption, ensure their slow release throughout the GIT, and improve their effects on fasted broiler chickens.

Adoptive transfer of ex vivo expanded SARS-CoV-2-specific cytotoxic lymphocytes: A viable strategy for COVID-19 immunosuppressed patients?

Cellular and humoral response to acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections is on focus of research. We evaluate herein the feasibility of expanding virus-specific T cells (VST) against SARS-CoV-2 ex vivo through a standard protocol proven effective for other viruses. The experiment was performed in three different donors' scenarios: (a) SARS-CoV-2 asymptomatic infection/negative serology, (b) SARS-CoV-2 symptomatic infection/positive serology, and (c) no history of SARS-CoV-2 infection/negative serology. We were able to obtain an expanded VST product from donors 1 and 2 (1.6x and 1.8x increase of baseline VST count, respectively) consisting in CD3 + cells (80.3% and 62.7%, respectively) with CD4 + dominance (60% in both donors). Higher numbers of VST were obtained from the donor 2 as compared to donor 1. T-cell clonality test showed oligoclonal reproducible peaks on a polyclonal background for both donors. In contrast, VST could be neither expanded nor primed in a donor without evidence of prior infection. This proof-of-concept study supports the feasibility of expanding ex vivo SARS-CoV-2-specific VST from blood of convalescent donors. The results raise the question of whether the selection of seropositive donors may be a strategy to obtain cell lines enriched in their SARS-CoV-2-specificity for future adoptive transfer to immunosuppressed patients.

Proposed global prognostic score for systemic mastocytosis: a retrospective prognostic modelling study.

BACKGROUND: Several risk stratification models have been proposed in recent years for systemic mastocytosis but have not been directly compared. Here we designed and validated a risk stratification model for progression-free survival (PFS) and overall survival (OS) in systemic mastocytosis on the basis of all currently available prognostic factors, and compared its predictive capacity for patient outcome with that of other risk scores. METHODS: We did a retrospective prognostic modelling study based on patients diagnosed with systemic mastocytosis between March 1, 1983, and Oct 11, 2019. In a discovery cohort of 422 patients from centres of the Spanish Network on Mastocytosis (REMA), we evaluated previously identified, independent prognostic features for prognostic effect on PFS and OS by multivariable analysis, and designed a global prognostic score for mastocytosis (GPSM) aimed at predicting PFS (GPSM-PFS) and OS (GPSM-OS) by including only those variables that showed independent prognostic value (p<0·05). The GPSM scores were validated in an independent cohort of 853 patients from centres in Europe and the USA, and compared with pre-existing risk models in the total patient series (n=1275), with use of Harrells' concordance index (C-index) as a readout of the ability of each model to risk-stratify patients according to survival outcomes. FINDINGS: Our GPSM-PFS and GPSM-OS models were based on unique combinations of independent prognostic factors for PFS (platelet count ≤100 × 109 cells per L, serum ß2-microglobulin ≥2·5 µg/mL, and serum baseline tryptase ≥125 µg/L) and OS (haemoglobin ≤110 g/L, serum alkaline phosphatase ≥140 IU/L, and at least one mutation in SRSF2, ASXL1, RUNX1, or DNMT3A). The models showed clear discrimination between low-risk and high-risk patients in terms of worse PFS and OS prognoses in the discovery and validation cohorts, and further discrimination of intermediate-risk patients. The GPSM-PFS score was an accurate predictor of PFS in systemic mastocytosis (C-index 0·90 [95% CI 0·87-0·93], vs values ranging from 0·85 to 0·88 for pre-existing models), particularly in non-advanced systemic mastocytosis (C-index 0·85 [0·76-0·92], within the range for pre-existing models of 0·80 to 0·93). Additionally, the GPSM-OS score was able to accurately predict OS in the entire cohort (C-index 0·92 [0·89-0·94], vs 0·67 to 0·90 for pre-existing models), and showed some capacity to predict OS in advanced systemic mastocytosis (C-index 0·72 [0·66-0·78], vs 0·64 to 0·73 for pre-existing models). INTERPRETATION: All evaluated risk classifications predicted survival outcomes in systemic mastocytosis. The REMA-PFS and GPSM-PFS models for PFS, and the International Prognostic Scoring System for advanced systemic mastocytosis and GPSM-OS model for OS emerged as the most accurate models, indicating that robust prognostication might be prospectively achieved on the basis of biomarkers that are accessible in diagnostic laboratories worldwide. FUNDING: Carlos III Health Institute, European Regional Development Fund, Spanish Association of Mastocytosis and Related Diseases, Rare Diseases Strategy of the Spanish National Health System, Junta of Castile and León, Charles and Ann Johnson Foundation, Stanford Cancer Institute Innovation Fund, Austrian Science Fund.

Targeted-Release Organic Acids and Essential Oils Improve Performance and Digestive Function in Broilers Under a Necrotic Enteritis Challenge.

An experiment was performed to evaluate the effect of four different microencapsulated blends of organic acids (OA) and nature-identical aromatic compounds (AC) on growth performance and gut health of broilers challenged with a recycled NE litter. A total of 600 one-day-old male Ross 308 broilers were randomly assigned to five treatments consisting of a basal diet (as negative control) supplemented with each of the tested microencapsulated blends: OA1 (malic and fumaric acid) + AC; 2.5 g/kg; OA2 (calcium butyrate+fumaric acid) + AC; 1.7 g/kg; MCFA (capric-caprylic; caproic and lauric acid) + AC; 2 g/kg; and MCFA + OA3(calcium butyrate + fumaric and citric acid) + AC; 1.5 g/kg. The AC used was the same for all treatments; including cinnamaldehyde, carvacrol, and thymol (8:1:1), as major compounds. Three tested blends enhanced growth performance by improving intestinal histomorphology (p < 0.001). The tested blends enhanced the abundance of some beneficial families such as Ruminococcaceae and Lachnospiraceae; while reducing that of harmful ones such as Enterobacteriaceae and Helicobacteraceae. A further dose-response experiment showed that 0.5 g/kg of the blend 2 and 2 g/kg of the blend 4 improved growth performance and intestinal histomorphology of chickens on d 42 and decreased fecal Enterobacteriaceae and C. perfringens counts. Similar effects to the previous experiment were observed for cecum microbiota.

Unravelling the Role of Rumen Microbial Communities, Genes, and Activities on Milk Fatty Acid Profile Using a Combination of Omics Approaches.

Milk products are an important component of human diets, with beneficial effects for human health, but also one of the major sources of nutritionally undesirable saturated fatty acids (SFA). Recent discoveries showing the importance of the rumen microbiome on dairy cattle health, metabolism and performance highlight that milk composition, and potentially milk SFA content, may also be associated with microorganisms, their genes and their activities. Understanding these mechanisms can be used for the development of cost-effective strategies for the production of milk with less SFA. This work aimed to compare the rumen microbiome between cows producing milk with contrasting FA profile and identify potentially responsible metabolic-related microbial mechanisms. Forty eight Holstein dairy cows were fed the same total mixed ration under the same housing conditions. Milk and rumen fluid samples were collected from all cows for the analysis of fatty acid profiles (by gas chromatography), the abundances of rumen microbiome communities and genes (by whole-genome-shotgun metagenomics), and rumen metabolome (using 500 MHz nuclear magnetic resonance). The following groups: (i) 24 High-SFA (66.9-74.4% total FA) vs. 24 Low-SFA (60.2-66.6%% total FA) cows, and (ii) 8 extreme High-SFA (69.9-74.4% total FA) vs. 8 extreme Low-SFA (60.2-64.0% total FA) were compared. Rumen of cows producing milk with more SFA were characterized by higher abundances of the lactic acid bacteria Lactobacillus, Leuconostoc, and Weissella, the acetogenic Proteobacteria Acetobacter and Kozakia, Mycobacterium, two fungi (Cutaneotrichosporon and Cyphellophora), and at a lesser extent Methanobrevibacter and the protist Nannochloropsis. Cows carrying genes correlated with milk FA also had higher concentrations of butyrate, propionate and tyrosine and lower concentrations of xanthine and hypoxanthine in the rumen. Abundances of rumen microbial genes were able to explain between 76 and 94% on the variation of the most abundant milk FA. Metagenomics and metabolomics analyses highlighted that cows producing milk with contrasting FA profile under the same diet, also differ in their rumen metabolic activities in relation to adaptation to reduced rumen pH, carbohydrate fermentation, and protein synthesis and metabolism.

Partial T Cell-Depleted Peripheral Blood Stem Cell Transplantation from HLA-Identical Sibling Donors for Patients with Severe Aplastic Anemia.

We analyzed the outcomes of 26 consecutive patients with acquired severe aplastic anemia (SAA) undergoing peripheral blood stem cell transplantation (PBSCT) with partial ex vivo T cell depletion with a targeted T cell dose from HLA-identical sibling donors. The median patient age was 37 years (range, 3 to 63 years). Four patients with uncontrolled pneumonia at the time of transplantation died, on days +1, +2, +21, and +26. All evaluable patients engrafted, with a median time to neutrophil recovery of 11 days (range, 10 to 14 days) and a median time to platelet recovery of 19 days (range, 8 to 53 days). Two patients had transient grade I acute graft-versus-host disease (GVHD) with skin involvement, but no patients developed grade II-IV acute GVHD. Two patients had mild skin chronic GVHD, and 1 patient had moderate chronic GVHD with ocular involvement. No relapse was observed after a median follow-up of 114 months (range, 4 to 233 months). The overall cumulative incidence of TRM at 10 years was 19%, whereas it was 5% for those with a Karnofsky Performance Status (KPS) score >60 at the time of transplantation. Disease-free survival, overall survival, and GVHD and relapse-free survival at 10 years were 81%, 81%, and 80%, respectively, for all patients and 95%, 95%, and 90%, respectively, for patients with a KPS score >60 at transplantation. Our data indicate that PBSCT with partial ex vivo T cell-depleted targeted cell dose grafts from an HLA-identical sibling donor is a feasible, safe, and effective approach to reduce GVHD and cure patients with SAA.

RNA Sequencing Analysis for the Identification of a PCM1/PDGFRB Fusion Gene Responsive to Imatinib.

The platelet-derived growth factor receptor ß (PDGFRB) gene translocations lead to a spectrum of chronic myeloid neoplasms, frequently associated with eosinophilia. Clinical heterogeneity is associated with a molecular one. Here, we report a novel case of a patient harboring a t(5;8)(q33;p22) translocation, resulting in the PCM1/PDGFRB fusion. Conventional cytogenetics and RNA sequencing were performed to identify the chromosomes and the genes involved in the rearrangement, respectively. This study shows that the combination of different strategies is pivotal to fine-tune the diagnosis and the clinical management of the patient. After 1 year of treatment with imatinib, the patient achieves hematological and molecular remission. We present an attractive strategy to identify novel and/or cryptic fusions, which will be relevant for clinicians dealing with the diagnosis of the patients with myelodysplastic syndrome/myeloproliferative diseases with atypical manifestations.

Impact of azacitidine on red blood cell alloimmunisation in myelodysplastic syndrome.

BACKGROUND: The incidence of alloimmunisation in myelodysplastic syndromes (MDS) during the era of supportive treatment ranges from 9 to 56%. However, it is unknown if the widespread use of hypomethylating agents has changed the risk of immunisation. The aim of this study is to evaluate the impact of azacitidine (AZA) therapy on red blood cell (RBC) alloimmunisation in transfused patients with MDS, myelodysplastic syndromes/myeloproliferative syndromes (MDS/MPS) and secondary acute myeloid leukaemia (AML). MATERIAL AND METHODS: We have analysed retrospectively all patients with MDS, MDS/MPS and secondary AML from MDS, who received their first transfusion in our hospital between January 1995 and December 2014. We have assessed the impact of age, sex, RBC and platelets units transfused, and AZA treatment on developing alloantibodies. RESULTS: In our study, the number of RBC units transfused increased the risk of developing alloantibodies. However aging and the treatment with AZA were associated with a lower rate of alloimmunisation. DISCUSSION: Patients with MDS, MDS/MPS and secondary AML who received treatment with AZA developed RBC antibodies at a lower rate than control group. We suggest that aging and immunosuppression due to AZA therapy could develop an immunological tolerance with a weak response to allotransfusions.

Negative impact on clinical outcome of the mutational co-occurrence of SF3B1 and DNMT3A in refractory anemia with ring sideroblasts (RARS).

The incidence of SF3B1 mutations in patients with RARS is high. Recently, it has been shown that SF3B1 and DNMT3A mutations overlap more often than expected, although it is not clear how this could affect the disease. We studied SF3B1 and DNMT3A in 123 RARS patients: 101 out of 123 samples (82%) had somatic mutations in SF3B1, and 13 of them (13%) showed a co-mutation (SF3B1mutDNMT3Amut). All co-mutated patients had a normal karyotype, and 12 of them (92%) were lower-risk patients (IPSS and IPSS-R). Despite their favorable profile, SF3B1mutDNMT3Amut patients showed a higher RBC transfusion dependency (92% versus 48%, p = .007), a shorter overall survival (OS) (median, 30 versus 97 months, p = .034), and a higher risk of progression to acute myeloid leukemia (AML) at 5 years (25% versus 2%, p = .023) than SF3B1mutDNMT3Awt patients. In conclusion, DNMT3A mutations are present in a significant proportion of SF3B1mut patients with a negative clinical impact.

Study of the S427G polymorphism and of MYBL2 variants in patients with acute myeloid leukemia.

Dysregulation of MYBL2 has been associated to tumorigenesis and the S427G polymorphism could induce partial inactivation of MYBL2, associating it with cancer risk. It has previously been shown that MYBL2 was over-expressed in some acute myeloid leukemias (AML), portending poor prognosis. However, to date no studies have investigated the S427G or other genetic variants of MYBL2 in AML. This study analyzed the S427G in 197 AML patients and 179 controls and screened the MYBL2 sequence in patients. In contrast to other studies in solid tumors, the S427G was not associated with the incidence of AML. This study detected four unannotated genetic alterations, of which the Q67X could be involved in MYBL2 dysfunction. Eight polymorphisms were identified, among which the rs73116571, located in a splicing region, was associated with higher incidence in AML and weaker MYBL2 expression, suggesting pre-disposition to AML. Additional functional studies should be performed to verify these genetic variations as possible targets in AML.

Evaluación del optimismo disposicional en pacientes en hemodiálisis y su relación con las complicaciones / Assessment of dispositional optimism in hemodialysis patients and its relation to complications

Introducción: El optimismo parece tener una influencia sobre la resistencia a la enfermedad y de la mejora de la salud en pacientes con problemas cardíacos, respiratorios y oncológicos. Sin embargo, son escasos los estudios en pacientes con enfermedad crónica renal en hemodiálisis; de ahí el interés de nuestro estudio. Objetivo: El objetivo de este estudio fue analizar la relación entre el grado de optimismo disposicional con las complicaciones y la calidad de vida percibida en un grupo de pacientes de hemodiálisis. Pacientes y Métodos: Se estudiaron 78 pacientes en HD, del servicio de Nefrología de Córdoba. Se realizó un estudio observacional analítico. Se evaluaron variables demográficas, tiempo en diálisis, índice de comorbilidad de Charlson, centro de diálisis, acceso vascular, optimismo disposicional mediante el LOT-R (a más puntuación más optimismo), calidad de vida relacionada con la salud mediante láminas COOP-WONCA (a más puntuación peor calidad de vida percibida) y número de ingresos en el año anterior. Resultados: La edad media fue 64.8±17.1 años. El 38.5% eran mujeres y el 61.5% hombres. La mediana del número de ingresos en el último año fue de 1 (rango: 0-10). La mediana del índice de comorbilidad fue 7 (rango 0-12). La media del LOT-R fue 20.76±4.8 y la mediana 21 (rango: 8-29). Se estableció la mediana del LOT-R, como punto de corte para clasificar a los pacientes como optimistas (≥21 de mediana) y pesimistas (<21). Al comparar ambos grupos (optimistas frente a pesimistas) se encontraron diferencias significativas para el COOP-WONCA (19.8±5.6 frente a 24.5±6.6, p<0.002). Sin embargo no se encontraron diferencias para el número de ingresos, ni para el índice de comorbilidad, entre ambos grupos. Conclusiones: No se encontró relación entre optimismo disposicional y número de ingresos, ni comorbilidad asociada. Sin embargo, los más optimistas tienen mayor de calidad de vida percibida relacionada con la salud (AU)

Introduction: Optimism seems to have an influence on the resistance to disease and on the improvement of the health in patients with cardiac, respiratory and cancer problems. However, there are few studies in patients with chronic renal disease on haemodialysis; hence the importance in our study. Objective: The aim of this study was to analyse the relationship between the degree of dispositional optimism with complications and perceived quality of life in a group of haemodialysis patients. Patients and Methods: 78 patients were studied in HD, Nephrology Unit of Córdoba. An observational study was conducted. Demographic variables, time on dialysis, Charlson comorbidity index, dialysis centre, vascular access, dispositional optimism by LOT-R (higher score, higher optimism), health-related quality of life was evaluated by COOP-WONCA sheets (highest score worse perceived quality of life) and number of admissions in the previous year. Results: The mean age was 64.8 ± 17.1 years. 38.5% were female and 61.5% male. The median number of admissions in the last year was 1 (range: 0-10). Median comorbidity index was 7 (range 0-12). The mean of the LOT-R was 20.76 ± 4.8 and the median 21 (range 8-29). Median LOT-R was established as a cut-off point to classify patients as optimistic (≥21 middle) and pessimistic (<21). When comparing both groups (optimistic vs. pessimistic) significant differences for the COOP-WONCA (19.8 ± 5.6 versus 24.5 ± 6.6, p <0.002) were found. However no differences, between both groups, were found for the number of admissions and for the comorbidity index. Conclusions: No relationship was found between dispositional optimism and number of admissions, neither comorbidity. However, the optimists had higher perceived health-related quality of life (AU)

Single-nucleotide polymorphism array-based karyotyping of acute promyelocytic leukemia.

Acute promyelocytic leukemia (APL) is characterized by the t(15;17)(q22;q21), but additional chromosomal abnormalities (ACA) and other rearrangements can contribute in the development of the whole leukemic phenotype. We hypothesized that some ACA not detected by conventional techniques may be informative of the onset of APL. We performed the high-resolution SNP array (SNP-A) 6.0 (Affymetrix) in 48 patients diagnosed with APL on matched diagnosis and remission sample. Forty-six abnormalities were found as an acquired event in 23 patients (48%): 22 duplications, 23 deletions and 1 Copy-Neutral Loss of Heterozygocity (CN-LOH), being a duplication of 8(q24) (23%) and a deletion of 7(q33-qter) (6%) the most frequent copy-number abnormalities (CNA). Four patients (8%) showed CNAs adjacent to the breakpoints of the translocation. We compared our results with other APL series and found that, except for dup(8q24) and del(7q33-qter), ACA were infrequent (≤3%) but most of them recurrent (70%). Interestingly, having CNA or FLT3 mutation were mutually exclusive events. Neither the number of CNA, nor any specific CNA was associated significantly with prognosis. This study has delineated recurrent abnormalities in addition to t(15;17) that may act as secondary events and could explain leukemogenesis in up to 40% of APL cases with no ACA by conventional cytogenetics.

Análisis comparativo entre hemodiafiltración en línea frente a hemodiafiltración con reinfusión endógena: parámetros clínico-técnicos e inflamatorios / A comparative analysis between online hemodia-filtration and hemodiafiltration with endogenous reinfusion: clinical, technical and inflammatory parameters

Introducción: El tratamiento con hemodiálisis produce «per se» un proceso inflamatorio crónico en el paciente. Diferentes técnicas alternativas como la hemodiafiltración «en línea», mejoran entre otros aspectos, este perfil inflamatorio. En esta misma línea de investigación están apareciendo nuevas técnicas convectivas que sobre el papel podrían mejorar a la hemodiafiltración «en línea». La técnica HFR-SUPRA o hemodiafiltración con reinfusión endógena utiliza un dializador con doble cámara más un cartucho de resina, en el que el propio líquido ultrafiltrado del paciente es reinfundido posteriormente a su regeneración en este cartucho de resina. El objetivo de nuestro estudio fue evaluar las diferencias entre la técnica de hemodiafiltración con reinfusión endógena y la hemodiafiltración «en línea», en cuanto a parámetros clínicos, inflamatorios y adecuación de la diálisis. Material y métodos: Se incluyeron 16 pacientes, de edad media 61,6±14,5 años, sometidos a las dos técnicas durante períodos de 8 semanas, según el siguiente esquema: hemodiafiltración con reinfusión endógena -1, hemodiafiltración "en línea" -1, hemodiafiltración con reinfusión endógena -2, hemodiafiltración «en línea» -2. Todos los pacientes estuvieron 8 semanas previas con hemodiálisis de alto flujo. Se recogieron parámetros técnicos (presión en las líneas arterial y venosa, flujo de sangre, litros tratados, volumen de infusión, necesidades de heparina), clínicos (presión arterial sistólica y diastólica pre y post-diálisis, peso seco, ganancia interdiálisis, incidencias durante la sesión) y de laboratorio (eliminación de β2m, hemoglobina, hematocrito, albúmina) y parámetros inflamatorios. También se analizó la duración del montaje, mediante la media de tiempo en minutos, de 10 sesiones seguidas con ambas técnicas. Resultados: No hubo diferencias entre ambas técnicas en los parámetros clínicos recogidos: presión arterial sistólica y diastólica pre y post-diálisis, peso seco, ganancia interdiálisis, incidencias durante la sesión. Tampoco hubo diferencias significativas en las necesidades de heparina ni en el Kt/V. Los valores de hemoglobina (11.46 gr/dl en ambas), reducción de β2m (70.14% vs 65.19%, p=0.6) y albúmina (3.60 vs 3.64, gr/dl p=0.4) no fueron diferentes. Se encontró una mayor presión en la línea venosa en la hemodiafiltración «en línea» (165.2 vs 149.9, p=0.017), junto a una mayor hemoconcentración observada de forma subjetiva por Enfermería, si bien el volumen de líquido de reposición es significativamente mayor con hemodiafiltración «en línea». El tiempo de montaje y cebado es mayor en hemodiafiltración con reinfusión endógena HFR (2.28 vs 4.82, p=0.012), aunque no se observa más dificultad en el montaje de la técnica. La sesión la hemodiafiltración con reinfusión endógena requirió menos intervención de enfermería, ya que presenta menos variabilidad en los parámetros relacionados con la hemoconcentración, generando una menor cantidad de alarmas. No hubo diferencias significativas en la PCR como marcador inespecífico de inflamación, sin embargo, en el laboratorio experimental se observó una disminución de la actividad de los CD14++ y CD16++ (monocitos pro-inflamatorios) en la hemodiafiltración con reinfusión endógena, y también presentó menos niveles de VEGF. Conclusiones: La hemodiafiltración con reinfusión endógena es una técnica que no plantea más dificultades técnicas que la hemodiafiltración «en línea» para enfermería, pues aunque el tiempo de montaje es mayor, requiere menor intervención posterior. No se encontraron diferencias en los parámetros técnicos y clínicos entre ambas técnicas. Sin embargo nuestros resultados preliminares muestran una disminución de la activación proinflamatoria de los monocitos en la hemodiafiltración con reinfusión endógena, con respecto a la hemodiafiltración «en línea» (AU)

Introduction: Hemodialysis (HD) itself produces a chronic inflammatory process in the patient. Alternative techniques as hemodiafiltration «online» (OL-HDF) improve, among other things, this inflammatory profile. Nowadays, in this research line, new convective techniques are emerging. These techniques could be better than the HDF-OL. Hemodiafiltration with endogenous reinfusion or HFR-SUPRA (HFR) uses a two-chamber dialyzer plus a resin cartridge. In this technique the patient ultrafiltered liquid is subsequently reinfused to its regeneration in this resin cartridge. The aim of our study was to evaluate the differences between the HFR and OL-HDF technique, in terms of clinical and inflammatory parameters and dialysis adequacy. Materials and methods: Sixteen patients were included with a mean age of 61.6 ± 14.5 years. These patients were subjected to the two techniques for periods of eight weeks by the following scheme: HFR-1, HDF-OL-1, HFR-2, HDF-OL-2. All patients underwent high-flux HD in the 8 weeks prior. Technical (pressure in the arterial and venous lines, blood flow, treated liter, the infusion volume, and needs of heparin), clinical (systolic and diastolic blood pressure pre and post-dialysis, dry weight, interdialytic weight gain, incidents during the hemodialysis session) and laboratory parameters (elimination of B2m, hemoglobin, hematocrit, albumin) and inflammatory parameters were collected. Results: There was no difference between the two techniques in the collected clinical parameters: systolic and diastolic pre and post-dialysis dry weight, interdialytic weight gain and incidents during the session. And even there were no significant differences in the needs of heparin or Kt / V. The hemoglobin values (both 11.46), B2m reduction (70.14% vs 65.19%, p = 0.6) and albumin (3.60 vs 3.64, p = 0.4) were not significantly different. It was found higher pressure in the venous line in the OL-HDF versus HFR (165.2 vs 149.9, p = 0.017), together with greater hemoconcentration subjectively observed by Nursing. However, the volume of replacement fluid is significantly higher with the OL-HDF technique. The assembly time and priming were higher in the HFR technique (2.28 vs 4.82, p = 0.012), although greater difficulty in the assembly of the technique was not observed. During the session, the HFR technique requires less nursing intervention because it presents less variability in parameters related to hemoconcentration, generating fewer alarms. There were no significant differences in the PCR as nonspecific marker of inflammation. However, in the HFR technique a decrease in the activity of CD14 ++ and CD16 ++ (pro-inflammatory monocytes) and lower levels of VEGF were observed. Conclusion: The HFR technique does not present more technical difficulties than the HDF-OL for nursing, for though the installation time is longer, requires less subsequent intervention. No significant differences in the technical and clinical parameters between the two techniques were found. However, our preliminary results show a decrease of the proinflammatory activation of monocytes in the HFR with respect to the OL-HDF technique (AU)

Adverse prognostic value of MYBL2 overexpression and association with microRNA-30 family in acute myeloid leukemia patients.

The MYBL2 gene encodes a transcription factor implicated in cell proliferation and maturation whose amplification or overexpression has been associated with different human malignancies, suggesting that it could be implicated in tumorigenesis. We analyzed MYBL2 expression and its prognostic value in 291 patients with de novo acute myeloid leukemia (AML) and we also evaluated its association with microRNAs 29 and 30 families. MYBL2 expression in AML patients was increased relative to CD34+ cells. Moreover, MYBL2 overexpression was associated with lower expression of miR-30a (P=0.024), miR-30b (P=0.021) and miR-30c (P=0.009). Multivariate analysis showed that MYBL2 expression was an independent factor for disease-free survival (HR 3.0, 95% CI 1.5-6.0, P=0.002) and cumulative incidence of relapse (HR 2.6, 95% CI 1.2-5.6, P=0.015) in patients with an intermediate-risk karyotype. In conclusion, our data showed that MYBL2 expression analysis could be useful to define subgroups of patients with poor prognosis.

WT1 isoform expression pattern in acute myeloid leukemia.

WT1 plays a dual role in leukemia development, probably due to an imbalance in the expression of the 4 main WT1 isoforms. We quantify their expression and evaluate them in a series of AML patients. Our data showed a predominant expression of isoform D in AML, although in a lower quantity than in normal CD34+ cells. We found a positive correlation between the total WT1 expression and A, B and C isoforms. The overexpression of WT1 in AML might be due to a relative increase in A, B and C isoforms, together with a relative decrease in isoform D expression.