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CD34 positivity has been considered as an adverse prognostic factor in acute myeloid leukemia (AML). Although nucleophosmin 1-mutated (NPM1m) AML is usually CD34 negative, this marker may be expressed at diagnosis or acquired at relapse in a variable number of cases. Our objective was to ascertain if CD34 expression has any influence on the general outcome of this form of acute leukemia. Analysis of clinical outcome (complete remissions, relapses, disease-free survival, and overall survival) was performed depending on the degree of expression of CD34 determined by flow cytometry, in 67 adult patients with NPM1m AML. CD34 expression did not have any influence on the variables analyzed whatever the percentage of blasts expressing this marker. In contrast to other forms of AML, CD34 expression is not an unfavorable prognostic factor in NPM1m AML, neither at diagnosis nor at relapse.

BACKGROUND Some copy number alterations (CNAs) have independent prognostic significance for adults with acute lymphoblastic leukemia (ALL). METHODS This study analyzed via multiplex ligation–dependent probe amplification the frequency and prognostic impact of CNAs of 12 genetic regions in 142 adolescents and adults with de novo precursor B-cell ALL. RESULTS The cyclin-dependent kinase inhibitor 2A/B (CDKN2A/B) deletion (59 of 142 or 42%) was the most frequent CNA, and it was followed by Ikaros family zinc finger 1 (IKZF1) losses (49 of 142 or 35%). IKZF1 deletions were more prevalent in Philadelphia chromosome (Ph)–positive ALL and were associated with advanced age and high white blood cell (WBC) counts. The multivariate analysis showed that advanced age and early B-cell factor 1 (EBF1) deletions were associated with chemotherapy resistance in both the whole series (hazard ratios, 0.949 and 0.135, respectively) and the Ph-negative subgroup (hazard ratios, 0.946 and 0.118, respectively). High WBC counts and focal IKZF1 deletions correlated with disease recurrence (hazard ratios, 1.005 and 1.869, respectively), whereas advanced age and CDKN2A/B losses influenced overall survival in both the whole series (hazard ratios, 1.038 and 2.545, respectively) and the Ph-negative subgroup (hazard ratios, 1.044 and 2.105, respectively). CONCLUSIONS Deletions of EBF1, IKZF1, and CDKN2A/B have an independent adverse prognosis for adolescents and adults with B-precursor ALL, and this suggests that these CNAs should be included in the initial risk assessment of ALL. Cancer 2015;121:3809–3817. © 2015 American Cancer Society.

Objective:To characterize the effect of the HIV-1 infection and antiretroviral treatment (ART) in the human memory B (MEB)-cell compartment.Design:A cross-sectional study was designed to analyze MEB cells of HIV-1(+) ART treated and ART-naive study participants, and uninfected individuals.Methods:Frequency and absolute counts of MEB cell subsets in blood were determined by multicolor flow cytometry. Spontaneous cell death and B-cell proliferative capacity was evaluated in vitro by cell culture and flow cytometry. Splenic function was determined by pitted erythrocytes quantification in HIV-1(+) ART-treated study participants.Results:HIV-1(+) ART-treated individuals did not show functional hyposplenism despite the lack of recovery IgM(+)IgD(+)CD27(+) marginal zone-like B cells. Moreover, two germinal center-dependent MEB cells subsets were also dysregulated in HIV-1(+) individuals: IgM(+)IgD(-)CD27(+) (IgM only) cells were increased, whereas the switched subset (IgM(-)IgD(-)) was reduced in viremic individuals. Althought ART restored the numbers of these populations; the switched MEB cells were enriched in CD27(-) cells, which showed the highest susceptibility to spontaneous cell death ex vivo. In addition, B cells from viremic individuals showed a poor response to B-cell receptor and toll-like receptor 9 stimulation that was circumvented when both stimuli were used simultaneously.Conclusion:B cells from HIV-1(+) study participants show a poor stimulation capacity, that may be bypassed by the proper combination of stimuli, and a dysregulated MEB cell pool that suggest an affectation of the germinal center reaction, only partially normalized by ART. Interestingly, hyposplenism does not explain the lack of recovery of the marginal zone-like B cells in ART-treated HIV-1(+) individuals.

The BCL2 gene is located in the 18q21 chromosome whose protein has antiapoptotic effects. The most common chromosomal abnormality resulting in overexpression of BCL2 is t(14;18)(q32;q21), which is ...

Acute myeloid leukemia (AML) with myelodysplasia-related changes is characterized by the presence of multilineage dysplasia (MLD), frequently related to high-risk cytogenetics and poor outcome. However, the presence of MLD does not modify the favorable prognostic impact of NPM1 mutation. The prognosis of patients with AML presenting marked dysplasia lacking high-risk cytogenetics and NPM1 mutation is uncertain. We evaluated the prognostic impact of MLD in 177 patients with intermediate-risk cytogenetics AML (IR-AML) and wild-type NPM1. Patients were categorized as MLD-WHO (WHO myelodysplasia criteria; n = 43, 24 %), MLD-NRW (significant MLD non-reaching WHO criteria; n = 16, 9 %), absent MLD (n = 80, 45 %), or non-evaluable MLD (n = 38, 22 %). No differences concerning the main characteristics were observed between patients with or without MLD. Outcome of patients with MLD-WHO and MLD-NRW was similar, and significantly worse than patients lacking MLD. The presence of MLD (66 vs. 80 %, p = 0.03; HR, 95 % CI = 2.3, 1.08-4.08) and higher leukocyte count at diagnosis was the only variable associated with lower probability of complete remission after frontline therapy. Concerning survival, age and leukocytes showed an independent prognostic value, whereas MLD showed a trend to a negative impact (p = 0.087, HR, 95 % CI = 1.426, 0.95-2.142). Moreover, after excluding patients receiving an allogeneic stem cell transplantation in first CR, MLD was associated with a shorter survival (HR, 95 % CI = 1.599, 1.026-2.492; p = 0.038). In conclusion, MLD identifies a subgroup of patients with poorer outcome among patients with IR-AML and wild-type NPM1.

Among other phenotypic markers, CD11b expression has been considered as an unfavorable prognostic factor, both in terms of overall survival (OS), disease-free survival (DFS), and attainment and duration of complete remissions (CRs) in adult patients with acute myeloid leukemia (AML). Recently, some groups have restricted its prognostic impact to poor prognostic karyotypic risk groups. The aim of this study was to retrospectively analyze the prevalence of CD11b and of CD56 expression in blast cells of 158 AML patients [excluding those with t(15;17)] stratified according to their cytogenetic risk and to correlate these phenotypic characteristics with OS, DFS, and CR. CD11b was more frequently expressed in intermediate and unfavorable cytogenetic prognostic groups (38.9 and 35.5 %, respectively) than in the favorable group (9.5 %). No differences were observed in CD56 expression according to the cytogenetic risk groups. When OS, DFS, and CR were analyzed according to these two markers, no statistical differences were recorded in any cytogenetic risk group. In conclusion, although CD11b was more frequently expressed in blast cells of patients with intermediate and unfavorable cytogenetic risk groups, this feature did not translate into different clinical outcome. Similarly, CD56 positivity did not have any influence on the prognosis of these patients.

Background aims The increasing scarcity of young related donors has led to the use of older donors for related allogeneic hematopoietic stem cell transplantation (HSCT). This study analyzed the influence of age on the results of mobilization of peripheral blood stem cells (PBSCs) in healthy donors as well as on the engraftment and outcome of HSCT. Methods A retrospective analysis from a single center was performed comparing the results of PBSC mobilization from related healthy donors according to their age. Results The study included 133 consecutive related donors. The median age was 50 years (range, 4–77 years); 70 (53%) donors were males, and 44 (33%) were >55 years old. All donors were mobilized with granulocyte colony-stimulating factor for 5 days. The peak CD34 + cell count in peripheral blood was higher in younger than in older donors (median, 90.5 CD34 + cells/μL [range, 18–240 CD34 + cells/μL] versus 72 CD34 + cells/μL [range, 20–172.5 CD34 + cells/μL], P = 0.008). The volume processed was lower in younger than in older donors (16,131 mL [range, 4424–36,906 mL] versus 18,653 mL [range, 10,003–26,261 mL], P = 0.002) with similar CD34 + cells collected (579.3 × 10 6 cells [range, 135.14 × 10 6 –1557.24 × 10 6 cells] versus 513.69 × 10 6 cells [range, 149.81 × 10 6 –1290 × 10 6 cells], P = 0.844). There were no differences in time to recovery of neutrophils and platelets or in the incidences of acute and chronic graft-versus-host disease, overall survival, non-relapse mortality and relapse incidence. Conclusions Donors >55 years old mobilized fewer CD34 + cells and required a greater volume to collect a similar number of CD34 + cells. The outcome of HSCT was not influenced by donor age. Donor age should not be a limitation for related allogeneic HSCT.

Tyrosine kinase inhibitors (TKI) have improved the management of patients with chronic myeloid leukemia (CML). However, a significant proportion of patients does not achieve the optimal response or are resistant to TKI. ABL1 kinase domain mutations have been extensively implicated in the pathogenesis of TKI resistance. Although deletion or insertion of nucleotides in BCR-ABL1 has rarely been described, we identified a CML patient with an already described 35 nucleotides insertion (BCR-ABL1(35INS)) of controversial significance, that confers resistance to imatinib but sensitivity to dasatinib.

Acute myeloid leukemia (AML) is a heterogeneous group of hematological neoplasms with distinct clinical and genetic features. Chromosomal abnormalities are detectable by conventional cytogenetic an...

It is unclear whether higher CD34 + cell doses infused for ASCT have any influence on survival or relapse in patients with lymphoma. We analyzed the correlation of infused CD34 + cell dose with relapse, survival, and hematopoietic recovery in 146 consecutive patients undergoing ASCT for lymphoma. Higher doses (>5 × 106/kg) were significantly correlated with earlier hematopoietic recovery, fewer infectious episodes, lower transfusion needs. No differences were observed in lymphoma outcomes (4-year relapse incidence of 38% [95%CI: 29%–48%] in the lower dose group versus 51% [95%CI: 30%–69%] in the higher dose group, 10-year OS probabilities of 58% [95%CI: 48%–68%] versus 75% [95%CI: 59%–91%], 10-year DFS probabilities of 47% [95%CI: 37%–57%] versus 42% [95%CI: 23%–61%], p = NS for all outcomes). In this series, a higher infused CD34 + cell dose did not correlate with survival or relapse but correlated with earlier hematopoietic recovery and lower resource consumption.

Strategies to improve the efficiency of bronchoalveolar lavage (BAL) are needed. We conducted a study to establish the diagnostic value of BAL in patients with hematological malignancies and pulmonary infiltrates.The correlation of cytologic and flow cytometric study of BAL with the microbiological findings and the clinical evolution was determined.Seventy BAL were performed and flow cytometric study was analyzed in 23 of them. Fifty-three patients did not present any adverse event attributable to BAL. Anti-infectious therapy was modified in 64 (91%) patients. T lymphocyte count0.3×10Our study confirms the clinical value of BAL for treatment decision making in patients with hematological malignancies and acute respiratory failure.

// Laura Palomo 1, 2 , Olga Garcia 3 , Montse Arnan 4 , Blanca Xicoy 3 , Francisco Fuster 1 , Marta Cabezon 3 , Rosa Coll 5 , Vera Adema 1 , Javier Grau 3 , Maria-Jose Jimenez 3 , Helena Pomares 4 , Silvia Marce 3 , Mar Mallo 1 , Fuensanta Milla 3 , Esther Alonso 4 , Anna Sureda 4 , David Gallardo 5 , Evarist Feliu 3 , Josep-Maria Ribera 3 , Francesc Sole 1 , Lurdes Zamora 3 1 MDS Research Group, Josep Carreras Leukaemia Research Institute, ICO-Hospital Germans Trias i Pujol, Universitat Autonoma de Barcelona, Badalona, Spain 2 Departament de Bioquimica i Biologia Molecular, Universitat Autonoma de Barcelona, Badalona, Spain 3 Hematology Service, ICO-Hospital Germans Trias i Pujol, Josep Carreras Leukaemia Research Institute, Universitat Autonoma de Barcelona, Badalona, Spain 4 Hematology Service, ICO-Hospital Duran i Reynals, Barcelona, Spain 5 Hematology Service, ICO-Hospital Josep Trueta, Girona, Spain Correspondence to: Lurdes Zamora, email: lzamora@iconcologia.net Keywords: chronic myelomonocytic leukemia, normal karyotype, gene mutations, targeted deep sequencing, prognostic factors Received: May 20, 2016 Accepted: July 01, 2016 Published: July 29, 2016 ABSTRACT Clonal cytogenetic abnormalities are found in 20-30% of patients with chronic myelomonocytic leukemia (CMML), while gene mutations are present in >90% of cases. Patients with low risk cytogenetic features account for 80% of CMML cases and often fall into the low risk categories of CMML prognostic scoring systems, but the outcome differs considerably among them. We performed targeted deep sequencing of 83 myeloid-related genes in 56 CMML patients with low risk cytogenetic features or uninformative conventional cytogenetics (CC) at diagnosis, with the aim to identify the genetic characteristics of patients with a more aggressive disease. Targeted sequencing was also performed in a subset of these patients at time of acute myeloid leukemia (AML) transformation. Overall, 98% of patients harbored at least one mutation. Mutations in cell signaling genes were acquired at time of AML progression. Mutations in ASXL1 , EZH2 and NRAS correlated with higher risk features and shorter overall survival (OS) and progression free survival (PFS). Patients with SRSF2 mutations associated with poorer OS, while absence of TET2 mutations ( TET2 wt) was predictive of shorter PFS. A decrease in OS and PFS was observed as the number of adverse risk gene mutations ( ASXL1 , EZH2, NRAS and SRSF2 ) increased. On multivariate analyses, CMML-specific scoring system (CPSS) and presence of adverse risk gene mutations remained significant for OS, while CPSS and TET2wt were predictive of PFS. These results confirm that mutation analysis can add prognostic value to patients with CMML and low risk cytogenetic features or uninformative CC.

Failure in mobilization of peripheral blood (PB) stem cells is a frequent reason for not performing hematopoietic stem cell transplantation (HSCT). Early identification of poor mobilizers could avoid repeated attempts at mobilization, with the administration of pre-emptive rescue mobilization.Data from the first mobilization schedule of 397 patients referred consecutively for autologous HSCT between 2000 and 2010 were collected. Poor mobilization was defined as the collection of2 × 10(6) CD34(+)cells/kg body weight (BW).The median age was 53 years (range 4-70) and 228 (57%) were males. Diagnoses were multiple myeloma in 133 cases, non-Hodgkin's lymphoma in 114, acute myeloid leukemia or myelodysplastic syndrome in 81, Hodgkin's lymphoma in 42, solid tumors in 17 and acute lymphoblastic leukemia in 10. The mobilization regimen consisted of recombinant human granulocyte-colony-stimulating factor (G-CSF) in 346 patients (87%) and chemotherapy followed by G-CSF (C + G-CSF) in 51 (13%). Poor mobilization occurred in 105 patients (29%), without differences according to mobilization schedule. Diagnosis, previous therapy with purine analogs and three or more previous chemotherapy lines were predictive factors for poor mobilization. A CD34(+)cell count in PB13.8/μL was enough to ensure ≥ 2 × 10(6) CD34(+)cells/kg, with high sensitivity (90%) and specificity (91%).The prevalence of poor mobilization was high, being associated with disease type, therapy with purine analogs and multiple chemotherapy regimens. The threshold of CD34(+) cell count in PB identified poor mobilizers, in whom the administration of immediate or pre-emptive plerixafor could be useful to avoid a second mobilization.

Resumen Fundamento y objetivo El uso creciente de tratamientos antineoplasicos ha provocado un aumento de la incidencia de leucemias agudas secundarias (LAS). El objetivo de este trabajo fue describir las caracteristicas de las LAS en un solo centro. Pacientes y metodo Se registraron 23 casos de LAS. Se analizaron los agentes citotoxicos utilizados, tiempo hasta el diagnostico, caracteristicas clinico-biologicas, tratamiento y pronostico. Resultados La edad mediana fue de 61 anos. Los agentes administrados en la neoplasia previa fueron: alquilantes (17 pacientes), inhibidores de la DNA topoisomerasa II (14), antitubulina (12), radioterapia (9, en 6 con quimioterapia) y antimetabolitos (6). La mediana de tiempo hasta la aparicion de la LAS fue 3 anos (extremos 1,2-15,8). Trece pacientes recibieron quimioterapia intensiva (3 se consolidaron con un trasplante de progenitores hematopoyeticos [TPH]) mientras que los 10 restantes recibieron tratamiento de soporte (mediana de supervivencia de 3 anos frente a 0,079 anos, p = 0,004). Conclusiones En este estudio, la LAS se asocio con la exposicion de agentes alquilantes, inhibidores de la topoisomerasa II y agentes antitubulina. La respuesta al tratamiento y el pronostico fueron malos, aunque la quimioterapia y el TPH podrian proporcionar una supervivencia mas prolongada.

Resumen Fundamento y objetivo : La linfocitosis B policlonal persistente (LBPP) es una entidad muy poco frecuente que se relaciona con el tabaquismo e incide especialmente en mujeres. Cursa con aumento de IgM serica, asociacion al haplotipo HLA-DR7, anomalias citogeneticas y multiples reordenamientos de IgH/BCL-2 . Todavia no esta clara su naturaleza premaligna o benigna. El objetivo de este trabajo fue analizar las caracteristicas de la LBPP con especial interes en su evolucion. Pacientes y metodo Se han estudiado retrospectivamente 35 LBPP de 5 hospitales catalanes. Se realizo una valoracion morfologica de las extensiones de sangre por los miembros del Grup Catala de Citologia Hematologica (GCCH) en un microscopio de 16 cabezales y se analizaron los datos clinicos y biologicos. Resultados La LBPP se presento, en la mayoria de los casos, como linfocitosis en mujeres fumadoras. El distintivo morfologico es la presencia de linfocitos de aspecto activado, en ausencia de enfermedades viricas recientes, y de linfocitos bilobulados y/o hendidos, y algunos con bolsillos nucleares observados por ultraestructura. En la mayoria de los casos estudiados se detecto: aumento policlonal de IgM, expresion del haplotipo HLA-DR7, anomalias cromosomicas como i(3)(q10) y multiples reordenamientos de IgH/BCL-2 . Con una mediana de seguimiento de 70,7 meses, 34 de los 35 pacientes permanecen asintomaticos y vivos, uno fallecio por un adenocarcinoma de pulmon y otro desarrollo un linfoma folicular, sin demostracion de relacion alguna entre este y la LBPP. Conclusiones La LBPP presenta un curso estable y asintomatico, y se acompana con frecuencia de alteraciones geneticas. Se desconoce si es una situacion premaligna, a semejanza de las gammapatias monoclonales de significado incierto. Por ello, es fundamental una correcta interpretacion de la linfocitosis y un seguimiento evolutivo.

Splenic marginal zone lymphoma (SMZL) is a B-cell lymphoproliferative disorder with characteristic clinical, immunophenotypic, cytological and histological features. Some karyotypic abnormalities have been related to this disorder and most of them are usually complex and difficult to define. The aim of present study was to characterize new chromosomal aberrations involved in this disease. We performed conventional banding cytogenetics and Spectral Karyotyping (SKY) technique in 23 patients diagnosed with SMZL having a complex karyotype among a series of 160 SMZL cases. Del(7)(q22-q32) and trisomy 3/3q were the most common chromosomal aberrations. In addition, new translocations involving chromosomes 3, 6, 8, 9, 12 and 14q32 region were detected.

In the era of highly active antiretroviral therapy (HAART), the prognosis for acquired immunodeficiency syndrome-related lymphomas (ARL) seems to be similar to that for aggressive B-cell lymphomas in human immunodeficiency virus (HIV)-negative patients. This improvement in prognosis might lead to a modification of the classic prognostic factors for ARL. We evaluated the prognostic factors for response and survival in a series of HIV-infected patients with systemic non-Hodgkin's lymphoma (NHL) in the HAART era. Forty patients with systemic NHL treated with a CHOP-based chemotherapy (cyclophosphamide, doxorubicin, vincristine, and prednisone) and HAART were studied. The main clinicopathologic and laboratory parameters were recorded in each case. Patients were scheduled to receive cycles of CHOP therapy, and all received granulocyte colony-stimulating factor. In addition, 9 patients received rituximab (375 mg/m2). The complete remission (CR) rate was 62.5% (n = 25). No prognostic factors influencing CR attainment were found. The 5-year disease-free survival (DFS) probability (95% confidence interval [CI]) was 73% (54%-92%). The median overall survival (OS) time was 69.17 months, and the 5-year OS rate (95% CI) was 51% (35%-67%). A disease stage of III to IV was the only parameter with prognostic influence on DFS. The factors influencing OS were an International Prognostic Index2, an Eastern Cooperative Ecology Group (ECOG) score2, and a disease stage of III to IV. Patients with an advanced stage had a lower OS probability in a multivariate analysis (odds ratio, 4.24; 95% CI, 1.24- 14.57). Advanced stage was the main prognostic factor predicting survival in ARL treated with CHOP and HAART.

Fundamento y objetivo Cuantificar la funcion esplenica de los pacientes controlados en el Servicio de Cirugia General y Digestiva del Hospital Universitari Germans Trias i Pujol (HUGTiP) desde 1985 hasta 2003 tras haber sufrido diferentes grados de lesion traumatica del bazo segun la clasificacion de la American Association for the Surgery of Trauma (AAST) del ano 1994 y relacionarla con el tratamiento recibido (no operatorio, esplenectomia total con o sin esplenosis y esplenectomia mas autotrasplante), con la finalidad de detectar disfunciones esplenicas que predispongan al desarrollo de la sepsis tras la esplenectomia. Pacientes y metodo Se ha realizado a 43 pacientes un estudio isotopico con gammagrafia esplenica «dinamica », estudio de «pits» de la membrana eritrocitaria (optica de Nomarsky) y de vacuolas submembranarias (microscopia electronica de transmision). Resultados El grupo de no operados presenta una funcion normal de fagocitosis y filtracion, con una mediana de velocidad de captacion esplenica de 3,46 Kcts/s2 (intervalo, 0,8-6,98). El porcentaje mediano de «pits» de membrana fue del 2% (intervalo, 0-8,8%); el numero de «pits» por hematies, de 0,03 (intervalo, 0-0,12), y el porcentaje de hematies con 1, 2, 3 y 4 «pits», del 1,6, el 0,4, el 0 y el 0%, respectivamente. El porcentaje mediano de hematies con vacuolas submembranarias fue el 2,55% (intervalo, 0-5,6%); el numero de vacuolas por hematie, de 0,03 (intervalo, 0-0,06), y el porcentaje de hematies con 1, 2, 3 y 4 vacuolas, del 2, el 0,2, el 0 y el 0%, respectivamente. En el grupo operado, la velocidad mediana de captacion esplenica fue de 0,08 Kcts/s2 (intervalo, 0-1,75; p Conclusiones La funcion esplenica de los pacientes con antecedentes de traumatismo esplenico tratados de forma conservadora es normal, independientemente del grado de la lesion. Ello refuerza la conveniencia de aplicar de entrada esta actitud a todos los protocolos para intentar evitar la sepsis tras la esplenectomia. En los casos tratados con esplenectomia, con o sin esplenosis, la funcion esplenica esta ausente o muy alterada, mientras que en los casos tratados con esplenectomia y autotrasplante esta parcialmente conservada.

Some copy number alterations (CNAs) have independent prognostic significance for adults with acute lymphoblastic leukemia (ALL).This study analyzed via multiplex ligation-dependent probe amplification the frequency and prognostic impact of CNAs of 12 genetic regions in 142 adolescents and adults with de novo precursor B-cell ALL.The cyclin-dependent kinase inhibitor 2A/B (CDKN2A/B) deletion (59 of 142 or 42%) was the most frequent CNA, and it was followed by Ikaros family zinc finger 1 (IKZF1) losses (49 of 142 or 35%). IKZF1 deletions were more prevalent in Philadelphia chromosome (Ph)-positive ALL and were associated with advanced age and high white blood cell (WBC) counts. The multivariate analysis showed that advanced age and early B-cell factor 1 (EBF1) deletions were associated with chemotherapy resistance in both the whole series (hazard ratios, 0.949 and 0.135, respectively) and the Ph-negative subgroup (hazard ratios, 0.946 and 0.118, respectively). High WBC counts and focal IKZF1 deletions correlated with disease recurrence (hazard ratios, 1.005 and 1.869, respectively), whereas advanced age and CDKN2A/B losses influenced overall survival in both the whole series (hazard ratios, 1.038 and 2.545, respectively) and the Ph-negative subgroup (hazard ratios, 1.044 and 2.105, respectively).Deletions of EBF1, IKZF1, and CDKN2A/B have an independent adverse prognosis for adolescents and adults with B-precursor ALL, and this suggests that these CNAs should be included in the initial risk assessment of ALL.

In December 2013 mutations were described in the calreticulin (CALR) gene in 67–71% and 56–88% of patients with JAK2 V617F and MPL negative essential thrombocythemia (ET) and primary myelofibrosis ...

Objectives: To compare, from a biological and clinical perspective, a significant group of patients with AML with inv(3)(q21q26.2) or t(3;3)(q21;q26.2) with another group of AML carrying different abnormalities of 3q at q21 or q26, the latter named as the AML abn(3q) group. Methods: We developed a national survey with the participation of 13 Spanish hospitals, and retrospectively reviewed (from 1990 to 2010) these subtypes of AML. Fifty-five patients were collected: 35 with AML inv(3)/t(3;3) and 20 with AML abn(3q). A data collecting page that included main features at diagnosis, therapeutic approach and response, and survival variables, was distributed and completed. Results: We did not find significant differences in sex, age, history of myelodysplastic syndrome or chemo-/radiotherapy, clinical presentation, WBC and platelet counts, hemoglobin level, blasts immunophenotype, serum lactatedehydrogenase, peripheral blood and bone marrow cellular dysplasia, and bone marrow biopsy findings. Although the association with monosomy 7 was significantly more frequent in AML inv(3)/t(3;3), this did not seem to influence outcome. The lack of response to the different modalities of treatment and the aggressive course of the disease were the standard in both cohorts of patients. Discussion: Although not yet recognized by the World Health Organization classification, our results are in agreement with the findings of other authors, who include both subsets of AML together in the same group of adverse prognosis. Conclusion: In an attempt to simplify and bound entities with similar genetic background and clinical behavior, it would be desirable to bring together both subgroups of AML in a single section.

CA 125 and CA 15.3 serum levels were measured at diagnosis, after treatment and at the time of recurrence in 200 consecutive patients (114 males, median age 56 years) with non-Hodgkin's lymphoma (NHL) to explore its usefulness in the evaluation of response to treatment and survival in patients with NHL compared to lactate dehydrogense (LDH) and beta2-microglobulin (beta2-M). Their association with the clinical - biologic parameters at diagnosis, response to treatment, event-free survival (EFS) and overall survival (OS) was analysed. Eighty-six patients (43%) had elevated CA 125 levels and 35 (17.5%) had elevated CA 15.3 levels at diagnosis. CA 125 was associated with advanced stage, lung, pleural or gastrointestinal tract involvement and CA 15.3 was correlated with advanced stage, bone involvement, aggressive histology and bulky disease. LDH had the highest predictive value for failure to achieve complete remission (P = 0.001). A shorter OS was associated with increased LDH (P0.0001), beta2-M (P = 0.013) and CA 125 (P = 0.025) whereas CA 15.3 was associated with a shorter EFS (P = 0.027). When elevated at diagnosis, CA 125 and CA 15.3 should be monitored during follow-up of patients with NHL.

Fundamento y objetivo Aunque cada vez son mas numerosas las indicaciones del trasplante de progenitores hematopoyeticos (TPH), este no siempre llega a efectuarse. El objetivo de este estudio ha sido analizar las causas por las que no se realiza el TPH en los enfermos remitidos a una unidad de trasplante. Pacientes y metodo Se registraron los datos correspondientes a la visita pretrasplante efectuada a 129 enfermos remitidos de forma consecutiva a una unidad de trasplante hematopoyetico entre diciembre de 1999 y noviembre de 2002 y se analizaron la frecuencia y las causas de falta de realizacion del TPH. Por ultimo, se compararon las caracteristicas de los pacientes trasplantados y no trasplantados. Resultados De los 129 pacientes, 119 fueron remitidos para la realizacion de un trasplante autogenico y 10 para un TPH alogenico. La edad media (DE) era de 46 (14) anos (intervalo, 13–69 anos) y 69 pacientes (53,5%) eran varones. Ciento dieciocho tenian hemopatias malignas y 11 tumores solidos. Sesenta y un enfermos se hallaban en respuesta completa y 68 en respuesta parcial. Hasta el momento del analisis se habian realizado 93 TPH en 90 pacientes (69,8%). Ochenta y un pacientes recibieron un TPH autogenico (un paciente recibio un doble TPH) y 10, un TPH alogenico (dos en un paciente). Dos pacientes (1,5%) permanecen en lista de espera para realizar un trasplante autogenico. En 37 pacientes (28,7%) no se llego a efectuar el TPH. Las causas incluyeron: recaida o progresion de la enfermedad en el tiempo en el que se planeo realizar el TPH en 12 (32,5%), aplazamiento o cambio en la decision terapeutica en 12 (32,5%), movilizacion insuficiente de progenitores hematopoyeticos en 9 (24%) y rechazo del TPH por parte del paciente en 4 (11%). Si se excluye el aplazamiento, la frecuencia de falta de realizacion del TPH fue del 19,4%. De los 37 pacientes en los que no se realizo el TPH, se habia efectuado previamente la recoleccion de progenitores hematopoyeticos a 21 (57%). Ambos grupos (trasplantados y no trasplantados) fueron comparables para las principales caracteristicas, excepto la edad, que fue significativamente mayor en el grupo de pacientes no trasplantados (51 [12] frente a 44 [14] anos; p Conclusiones En una proporcion significativa de pacientes el TPH programado no llega a realizarse. Las causas mas frecuentes son la progresion de la enfermedad y la mala movilizacion de progenitores hematopoyeticos. Un motivo adicional para no realizar el TPH es el aplazamiento de la indicacion, ya sea programado o bien debido a la aparicion de nuevas estrategias terapeuticas.

Dear Editor, Chronic myelomonocytic leukemia (CMML) has been classified into a new category of myelodysplastic/myeloproliferative diseases (MDS/MPN) according to the last WHO classification of myeloid malignancies [1]. However, some patients with CMML show proliferative features (MP-CMML) such as leukocytosis and organomegaly, making them clinically and biologically similar to myeloproliferative neoplasms (MPN). At biological level, JAK2V617F mutations in MP-CMML have been described at a lower frequency than in MPN [2]. Recently, CALR gene mutations have been described in 67–88 % JAK2V617F and MPL-negative MPN; these cases also have a good prognosis [3–7]. The role of CALR mutations in CMML is not well known, and the few studies carried out so far have not focused on MP subtype [3, 4, 8]. The objective of this study was to characterize the frequency, type, and prognostic implications of CALR mutations in MP-CMML. CALR exon 9 mutations were studied in 30 patients (22 males, median age 66 years) diagnosed of CMML according toWHO classification [1], and all of them belonged to MP-CMML according to the French-American-British Co-operative Leukaemia Group (Table 1). All patients provided informed consent for sample collection. Biological analyses were carried out in accordance with the Declaration of Helsinki. Mutation analysis was performed as previously described [3]. An informative result was obtained in 29/30 patients, and no CALR exon 9 mutation was found. Although a small percentage of mutated MP-CMML patients was expected [4], no mutations were detected in the samples studied. These results are in concordance with those of Klampfl T et al. [3] and Hou et al. [8], who found no CALR mutations in 64 and 48 CMML patients, respectively (subtype not specified), but they do differ from the results of Nangalia et al., who found 1/33 CMML mutated patient (subtype not specified) [4]. There is also another study [9] of other MDS/ MPN groups (atypical CML and unclassifiable MDS/MPN) where CALR mutations were not seen in either entity (n=30). Therefore, outside essential thrombocythemia and primary myelofibrosis, CALR mutations are only found in (1) a few patients with MDS/MPN, most of them defined as having refractory anemia with ring sideroblasts associated with marked thrombocytosis, even though the percentage described is very variable depending on the series (0 to 12.5 %) [3, 4, 10] and (2) a few MDS patients, mainly with refractory anemia with ring sideroblasts, refractory anemia, and refractory anemia with excess blasts [3, 4, 8, 10]. This observation could indicate a relationship between CALR mutations and the thrombocytosis phenotype within myeloid neoplasms, suggesting that calreticulin mutations primarily affect the biology of megakaryocytes [11]. L. Zamora (*) :M. Cabezon : S. Marce : L. Palomo : F. Milla : E. Feliu : B. Xicoy ICO Badalona, Hospital Germans Trias i Pujol, Institut de Recerca contra la Leucemia Josep Carreras, Badalona, Spain e-mail: lzamora@iconcologia.net

Combined highly active anti-retroviral therapy (HAART) with protease and reverse transcriptase inhibitors has modified the natural history of opportunistic infections and neoplasms in human immunodeficiency virus (HIV)-infected patients. We analysed the influence of HAART on the response to treatment and survival in a series of 58 patients with acquired immune deficiency syndrome (AIDS)-related non-Hodgkin's lymphoma (NHL) treated with CHOP (cyclophosphamide, hydroxydoxorubicin, vincristine and prednisone). Two groups of patients were included: (i) forty-one patients diagnosed with NHL between 1988 and 1996 who were not treated with HAART; (ii) seventeen patients diagnosed since 1996, who were receiving or commenced HAART when NHL was diagnosed. The response rate to CHOP was higher in group 2 (13 out of 17 cases; 75%) than in group 1 (14 out of 41 cases; 34%) (P = 0.003). The 2-year probability of event-free survival (EFS) [95% confidence interval (CI)] for group 1 was 0.5 (0.24-0.74), whereas for group 2 it was 0.85 (0.61-0.90) (P = 0.024). The lymphoma-free survival (LFS) was also significantly different for both groups (2-year LFS probability 0.53 vs. 1.0, P = 0.04). The median (95% CI) overall survival (OS) for group 1 was 7 months (range, 3-10.8 months), whereas it was not reached in group 2 (P = 0.0015). In the multivariate analysis for remission attainment, the only variables with a higher probability to achieve complete remission (CR) were HAART (P = 0.01) and International Prognostic Index score 1 (P = 0.02). The only statistically significant variable in the multivariate analysis for EFS was HAART (P = 0.049) and the variables with prognostic value for OS in the multivariate analysis were B symptoms (P = 0.01) and HAART (P = 0.003). Patients with AIDS-related NHL treated with CHOP and HAART had a higher CR rate than those treated only with CHOP. In this study, HAART was an independent prognostic factor for CR, OS and EFS in patients with AIDS-related NHL.

Fundamento y objetivo Se describen 6 casos de osteoporosis como primera manifestacion de una mastocitosis sistemica diagnosticados en un periodo de 6 anos. Resultados Se incluyo a 3 mujeres y 3 varones con edades entre 47 y 66 anos. El intervalo entre el diagnostico de osteoporosis y el de mastocitosis sistemica oscilo entre 0,5 y 17 anos; 3 presentaban afeccion cutanea; 2, hematologica y 2, gastrointestinal, y los valores de histami-nuria estaban elevados en los 6 casos; 4 pacientes tenian fracturas. Se inicio tratamiento con bisfosfonatos. A los 3 anos, el valor de densidad mineral osea (DMO) lumbar mejoro en los 5 pacientes evaluados. A pesar del aumento de DMO, 2 de ellos tuvieron nuevas fracturas vertebrales, por lo que se inicio tratamiento con teriparatida. Conclusiones La osteoporosis como primera manifestacion de una mastocitosis sistemica es muy poco frecuente. Suelen ser osteoporosis graves. Aunque los farmacos antirresortivos mejoran la masa osea en estos pacientes, continua habiendo riesgo de fracturas.

Background: Plasma cell leukemia (PCL) is an uncommon and aggressive disease caused by the clonal proliferation of atypical plasma cells with phenotypical abnormalities similar to those seen in multiple myeloma (MM), although at different rates. Here we report a case of IgD PCL with a very unusual CD138-/CD19+/CD4+ phenotype Methods: Peripheral blood and bone marrow samples from a 37 year old patient afflicted by an aggressive plasma cell dyscrasia were examined and analyzed by conventional morphology, flow cytometry and immunohistochemistry. Results: Analysis of peripheral blood fulfilled criteria for PCL (more than 20% and more than 2x10e9 cells/L). However, flow cytometry and immunohistochemistry phenotyping revealed that the cells were CD138-/CD38+/CD19+/CD4+/CD56-/CD117- Conclusion: PCL is diagnosed on peripheral blood smear. Immunophenotyping is a tool that can be helpful in diagnosing difficult cases but its atypical findings should not prevent the appropriate PCL diagnosis in clinically and morphologically unquestionable cases. © 2014 Clinical Cytometry Society.

Background: Acute T-cell leukemia lymphoma (ATLL) tumor cells generally express CD2/CD3/CD5, but lack CD7. These T cells are usually CD4+CD8- and strongly express CD25, although some variability in this basic pattern may be found. Here we report a case with a very unusual CD1a positive phenotype. Methods: Samples from peripheral blood, bone marrow aspirate, lymph node and CSF obtained from a 45 year old male patient with a T-cell lymphoproliferative disorder were immunophenotyped by multiparametric flow cytometry Analysis of HTLV-I genome integration in tumoral cells was performed by PCR. Results: Neoplastic T cells were cCD3, CD2/CD5/CD30/CD25 and CD1a positive, but CD3/CD7/CD4/CD8/CD34/CD10/TdT negative. Serology and integration of HTLV-I were positive. Conclusion: To the best of our knowledge CD1a expression has not been previously described in this entity. Its detection raised the differential diagnosis with acute T lymphoblastic leukemia. The rest of the phenotypic markers, the morphology of the neoplastic cells, and the demonstration of HTLV-I genome integration provided the final diagnosis. © 2013 Clinical Cytometry Society.

In up to 5–15% of studies of lymphoproliferative disorders (LPD), flow cytometry (FCM) or immunomorphologic methods cannot discriminate malignant from reactive processes. The aim of this work was to determine the usefulness of PCR for solving these diagnostic uncertainties. We analyzed IGH and TCRγ genes by PCR in 106 samples with inconclusive FCM results. A clonal result was registered in 36/106 studies, with a LPD being confirmed in 27 (75%) of these cases. Specifically, 9/9 IGH clonal and 16/25 TCRγ clonal results were finally diagnosed with LPD. Additionally, two clonal TCRγ samples with suspicion of undefined LPD were finally diagnosed with T LPD. Although polyclonal results were obtained in 47 of the cases studied (38 IGH and nine TCRγ), hematologic neoplasms were diagnosed in 4/38 IGH polyclonal and in 1/9 TCRγ polyclonal studies. There were also 14 PCR polyclonal results (four IGH, 10 TCRγ), albeit nonconclusive. Of these, 2/4 were eventually diagnosed with B-cell lymphoma and 3/10 with T-cell LPD. In eight IGH samples, the results of PCR techniques were noninformative but in 3/8 cases a B lymphoma was finally confirmed. We concluded that PCR is a useful technique to identify LPD when FCM is inconclusive. A PCR clonal B result is indicative of malignancy but IGH polyclonal and nonconclusive results do not exclude lymphoid neoplasms. Interpretation of T-cell clonality should be based on all the available clinical and analytical data. © 2013 International Clinical Cytometry Society

Background: Acute T-cell leukemia lymphoma (ATLL) tumor cells generally express CD2/CD3/CD5, but lack CD7. These T cells are usually CD4+CD8- and strongly express CD25, although some variability in this basic pattern may be found. Here we report a case with a very unusual CD1a positive phenotype. Methods: Samples from peripheral blood, bone marrow aspirate, lymph node and CSF obtained from a 45 year old male patient with a T-cell lymphoproliferative disorder were immunophenotyped by multiparametric flow cytometry Analysis of HTLV-I genome integration in tumoral cells was performed by PCR. Results: Neoplastic T cells were cCD3, CD2/CD5/CD30/CD25 and CD1a positive, but CD3/CD7/CD4/CD8/CD34/CD10/TdT negative. Serology and integration of HTLV-I were positive. Conclusion: To the best of our knowledge CD1a expression has not been previously described in this entity. Its detection raised the differential diagnosis with acute T lymphoblastic leukemia. The rest of the phenotypic markers, the morphology of the neoplastic cells, and the demonstration of HTLV-I genome integration provided the final diagnosis. © 2013 Clinical Cytometry Society.

In up to 5-15% of studies of lymphoproliferative disorders (LPD) flow cytometry (FCM) or immunomorphologic methods cannot discriminate malignant from reactive processes. The aim of this work was to determine the usefulness of PCR for solving these diagnostic uncertainties. We analyzed IGH and TCRγ genes by PCR in 106 samples with inconclusive FCM results. A clonal result was registered in 36/106 studies, with a LPD being confirmed in 27 (75%) of these cases. Specifically, 9/9 IGH clonal and 16/25 TCRγ clonal results were finally diagnosed with LPD. Additionally, 2 clonal TCRγ samples with suspicion of undefined LPD were finally diagnosed with T LPD. Although polyclonal results were obtained in 47 of the cases studied (38 IGH and 9 TCRγ), hematologic neoplasms were diagnosed in 4/38 IGH polyclonal and in 1/9 TCRγ polyclonal studies. There were also 14 PCR polyclonal results (4 IGH, 10 TCRγ), albeit non-conclusive. Of these, 2/4 were eventually diagnosed with B-cell lymphoma and 3/10 with T-cell LPD. In 8 IGH samples the results of PCR techniques were non-informative but in 3/8 cases a B lymphoma was finally confirmed. We concluded that PCR is a useful technique to identify LPD when FCM is inconclusive. A PCR clonal B result is indicative of malignancy but IGH polyclonal and non-conclusive results do not exclude lymphoid neoplasms. Interpretation of T-cell clonality should be based on all the available clinical and analytical data. © 2013 Clinical Cytometry Society.

Background & Objective: Acute Lymphoblastic Leukemia (ALL) is an aggressive neoplasia characterized by a high genetic heterogeneity both at diagnosis and at relapse. Due to the high incidence of relapse in adults and the dismal prognosis beyond recurrence, diagnosis and relapse samples of adult ALL patients were carefully analyzed in order to identify genetic alterations related with drug resistance and disease progression. Patients & Methods: Paired diagnosis-relapse bone marrow samples from 5 adult B-cell precursor ALL (B-ALL) patients were analyzed (Ph+ ALL [n=2], normal karyotype [n=1], t(1;19)(q23;p13) [n=1] and t(8;13)(p21-22;q12) [n=1]). Copy Number Alterations (CNA) were studied with Multiplex Ligation-dependent Probe Amplification (MLPA, kits P-335 and P-202 from MRC-Holland, Amsterdam, Netherlands) and Affymetrix CytoScan HD arrays (Affymetrix, Santa Clara, USA). In the array analyses, only the CNA that encompassed at least 25 markers were considered significant. Results: Regarding karyotype, 2 patients (1 Ph+ and 1 t(1;19) at diagnosis) showed the same chromosomal translocations within a complex karyotype at relapse. On the contrary, the other Ph+ patient showed a normal karyotype at relapse, while 2 patients did not experience any karyotypic change. Regarding immunophenotype, 3/5 patients showed changes on antigen expression from diagnosis to relapse such as expression of markers of immaturity (CD34, TdT positivity and CD38 negativity), loss of lymphoid markers (CD20 and CD22) and/or acquisition of myeloid markers (CD33 and CD66c). Concerning CNA, all relapse samples were genetically related to the diagnosis clone (common clonal origin). All relapsed populations lost CNA detected at diagnosis and/or acquired new CNA but retained some of the CNA showed at diagnosis revealing clonal evolution from ancestral clones. CNA in B-ALL key genes involved in lymphoid development (IKZF1, PAX5, EBF1,VPREB1 and BLNK), proliferation (CDKN2A/B, RB1, CRLF2, C-MYC and ERG), apoptosis (BTG1, TP53 and ATM), hematopoiesis transcription factors (ETV6 and MLL) and histone modifications (KDM6A) were detected, among others. Losses in 9p were the most recurrent event both at diagnosis and at relapse. CDKN2A/B deletions were observed in all relapse samples (3/5 in homozygosis) while PAX5 deletions were present in 4/5 relapsed cases. Interestingly, all relapse samples showed CNA favoring the activation and/or the transcription of proteins involved in the Akt/C-MYC signaling pathway. Another common feature (4/5 patients) were CNA affecting genes involved in drug transport such as several ABC transporter genes and genes related to drug resistance such as PRKDC and RUNX1T1 (in 3/4 of the cases, the CNA appeared exclusively at relapse or were already present at diagnosis and increased their frequency at relapse). CNA in genes that may confer stem cell characteristics (EGR1 and USP16) were another recurrent event at relapse (3/5 samples, 2 of them were not present at diagnosis). CNA affecting the X/Y PAR1 region (CRLF2, CSF2RA and IL3RA) or VPREB1 at 22q11.22 were detected in 3/5 relapse samples, respectively. An important apoptosis cluster at 11q21q24.2 (BIRC2/3, CASP1/4/5/12, hsa-miR-34b/c, ATM and BTG4) was lost in 2/5 relapse samples (one of them was not detected at diagnosis and the other increased its frequency at relapse). Finally, ETV6 deletion (12p13.2) and duplication of Xq26.2q28 (containing ABCD1, BCAP31 and genes coding for several cancer/testis antigens) were observed in 2 relapse samples. Conclusions: SNP arrays analysis of paired B-cell precursor ALL samples at diagnosis and at relapse allows the identification of genetic alterations potentially related with ALL progression. The systematic analysis of relapse samples could contribute to the identification of specific genetic targets with potential therapeutic impact for each patient (personalized medicine). Disclosures Martínez-López: Novartis: Honoraria, Speakers Bureau. Sole:Celgene: Membership on an entity's Board of Directors or advisory committees.

Background and objectives Tyrosine kinase inhibitors (TKI) have improved the management of patients with chronic myeloid leukemia (CML). However, a significant proportion of patients do not achieve the optimal response or are resistant to TKI. ABL kinase domain mutations have been extensively implicated in the pathogenesis of TKI resistance. Treatment with second-generation TKI has produced high rates of hematologic and cytogenetic responses in mutated ABL patients. The aim of this study was to determine the type and frequency of ABL mutations in patients who were resistant to imatinib or had lost the response, and to analyze the effect of second-generation TKI on their outcome. Patients and methods The presence of ABL mutations in 45 CML patients resistant to imatinib was evaluated by direct sequencing and was correlated with the results of the cytogenetic study (performed in 39 cases). The outcome of these patients after therapy with nilotinib or dasatinib was analyzed. Results ABL mutations were detected in 14 out of 45 resistant patients. Patients with clonal cytogenetic evolution tended to develop mutations more frequently than those without clonal evolution. Nine out of the 15 patients with ABL mutation responded to a treatment switch to nilotinib (n = 4), dasatinib (n = 2), interferon (n = 1) or hematopoietic stem cell transplantation (n = 2). Conclusion The frequency of ABL mutations in CML patients resistant to imatinib is high and is more frequent among those with clonal cytogenetic evolution. The change to second-generation TKI can overcome imatinib resistance in most of the mutated patients.

Fundamento Valorar la promocion de la donacion desangre realizada en el entorno de los pacientes quereciben un trasplante de progenitores hematopoyeticos(TPH) y compararla con la promocion que se realizaen el entorno del resto de pacientes ingresados. Pacientes y metodo El hematologo y el promotor delBanco de Sangre informaron de la probabilidad detransfusion al paciente candidato a TPH. Se efectuouna seleccion de los posibles donantes para ladonacion de sangre total o de plaquetas. En el restode los pacientes ingresados la promocion se efectuomediante la visita del promotor al paciente transfundidoo con solicitud de transfusion, sin intervenciondel medico que atendia al paciente. Resultados Entre enero de 2000 y mayo de 2001se realizaron 48 visitas a pacientes candidatos aTPH: 41 (85%) aportaron 453 posibles donantes,306 donaron sangre, 77 plaquetas y 82 fueron excluidos.Se obtuvieron 484 donaciones, 389 desangre total y 105 de plaquetas. Un 25% (98 donantes)volvio a donar espontaneamente. Entre elresto de los pacientes ingresados, durante el ano1998 se realizaron 1.950 entrevistas, consiguiendocolaboracion en 533 (27,3%) de los casos (p Conclusiones La implicacion del hematologo en lapromocion de la donacion a partir del entorno de lospacientes ingresados repercute en un aumento de latasa de donaciones y en la fidelidad de las mismas.

Primary gynecologic myeloid sarcomas are rare, and their diagnosis is often difficult. Differential diagnosis includes lymphomas and carcinomas of the gynecologic tract. We report the clinical, morphological, immunohistochemical and cytogenetic features of two cases of chloromas of the female genital tract, which progressed to acute myeloid leukemia in spite of aggressive therapy.

Primary lymphomas of the conjunctiva are extremely infrequent and usually belong to extranodal marginal zone B-cell lymphoma of MALT or MALT lymphoma. Radiotherapy with lens shielding is one of the most employed therapeutic options. Four patients with MALT lymphomas of the conjunctiva with complete and maintained response to radiotherapy are reported. The effectiveness as well as the lack of significant toxicity of radiotherapy are emphasized.

A proposal for a scoring system in the diagnosis of chronic lymphoproliferative diseases other than CLL has been recently published in Cytometry Part B. The authors apply this score for deciding whether or not FISH evaluation for the detection of IGH/CCND1 rearrangements must be performed to exclude Mantle Cell Lymphoma (MCL). In their validation series, no MCL scored

In this short report we describe a patient with human parvovirus B19 (HPV B19)-induced transient pancytopenia. Parvovirus virions were seen by electron microscopy in both erythroid and granulocytic precursors. Erythroid cells are not the only targets in these cases. We draw attention to this disorder so that physicians involved with hematological disorders and transplantation be more aware of this infection.

To the Editor Although the unequivocal diagnosis of acute promyelocytic leukemia (APL) is established by the finding of t(15;17) and/or the PML / RARA fusion gene, cytogenetic and molecular techniques are time-consuming. Instead, flow cytometry (FC) allows a prompt diagnosis of this form of leukemia and the rapid institution of the specific therapy. Several studies have tried to identify the panel of markers that better characterize this entity. In a series of 149 patients with APL, Dong et al1 found as constant the triple negativity for HLA-DR, CD11b, and CD11c. Similar results were obtained in a shorter study that analyzed 16 cases of APL,2 in which 100% of the patients were negative for CD34, CD14, and CD11b (CD11c was not studied). We hereby share a different experience in a similar study. The expression of CD34, CD11b, and DR, among other markers, was analyzed by FC in a retrospective series (years 2008–2013) of 18 patients (7 …

Fundamento Analizar la etiologia, los metodos diagnosticos y la respuesta al tratamiento en 30 episodios de neumonia diagnosticados en 17 pacientes con leucemia linfatica cronica (LLC) entre 1995 y 2000. Pacientes y metodo En cada episodio se registraron los siguientes parametros: edad, sexo, tratamiento de la LLC, profilaxis antiinfecciosa, granulocitopenia, cociente de linfocitos CD4 y CD8, hipogammaglobulinemia, tipo de neumonia (intra o extrahospitalaria), localizacion, insuficiencia respiratoria, necesidad de ventilacion mecanica, tratamiento antimicrobiano y respuesta. Se realizaron hemocultivos, cultivo de esputo, fibrobroncoscopia y deteccion de antigenos en orina (Legionella pneumophila serogrupo 1, galactomanano y Streptococcus pneumoniae). Resultados La edad mediana de la serie fue de 60 anos (limites, 50-86); 12 eran varones. La combinacion de clorambucilo y prednisona fue el tratamiento mas utilizado para la LLC (13 pacientes) seguido de la fludarabina (8 casos). Existia granulocitopenia en 14 episodios, habia hipogammaglobulinemiaen 22 y el cociente de linfocitos CD4 y CD8 fue inferior a uno en 8 de 14 determinaciones. Se establecio la etiologia de las neumonias en 16 episodios (53%). La fibrobroncoscopia fue la prueba con mayorrentabilidad diagnostica (83%), seguida de los hemocultivos (38%). Dos pacientes fueron diagnosticadosen la autopsia de aspergilosis pulmonar. El neumococo fue el germen aislado con mayor frecuencia(5) seguido de Pseudomonas aeruginosa (4), Pneumocystis carinii (2) y Aspergillus fumigatus (2). Delos 2 pacientes con neumocistosis uno habia recibido fludarabina y el otro glucocorticoides de formaprolongada. Diez pacientes (30%) fallecieron a causa de los siguientes germenes: P. aeruginosa (3),P. carinii (2), A. fumigatus (2), Mycobacterium xenopi (1) y germen no identificado (2). Conclusiones En esta serie de pacientes con LLC la tasa global de diagnostico etiologico de las neumonias fue aceptable. El germen mas frecuente fue el neumococo. Las neumonias por microorganismos oportunistas se relacionaron con la administracion de fludarabina o el tratamiento prolongado con glucocorticoides y tuvieron una elevada mortalidad.

The growing use of antineoplastic treatments has led to an increase in the incidence of therapy-related leukemias (TRL). The objective was to describe the characteristics of TRL.Twenty-three cases of TRL were registered. Chemotherapeutic agents used for the first tumor, time interval, clinical and biological characteristics, treatment and prognosis of the TRL were analyzed.Median age was 61 years. Cytotoxic agents used in previous neoplasm consisted of alkylating agents (17 patients), inhibitors of DNA topoisomerase II (14), antitubulin agents (12), radiotherapy (9, in 6 with radiotherapy) and antimetabolites (6). Median time from diagnosis of the first neoplasm to TRL was 3 years (range 1.2-15.8). Thirteen patients received intensive chemotherapy [with stem cell transplantation (SCT) in 3] and the other 10 received symptomatic treatment (median survival 3 years versus 0.079 years, P=0.004).In this study, TRL were associated with exposure to alkylating agents, antitubulin agents and topoisomerase II inhibitors. Response to treatment and prognosis were poor, although chemotherapy and SCT may prolong survival.

We conducted a retrospective collaborative study to cytogenetically characterize splenic marginal zone lymphoma (SMZL) and ascertain the prognostic value of chromosomal aberrations. Of 330 cases, 72% displayed an aberrant karyotype, 53% were complex, and 29% had a single aberration. The predominant aberrations were gains of 3/3q and 12q, deletions of 7q and 6q and translocations involving 8q/1q/14q. CD5 expression was detected in 39 of 158 cases (25%). The cytogenetic makeup of the CD5+ group differed significantly from that of the CD5− group. Cases with unmutated IGHV were significantly associated with deletions of 7q and TP53. A strong association was noted between usage of the IGVH1-2 and deletion 7q, 14q alterations, and abnormal karyotype. On univariate analysis, patients with more than or equal to 2 aberrations, 14q alterations, and TP53 deletions had the shortest survival; 7q deletion did not affect survival. On multivariate analysis, cytogenetic aberrations did not retain prognostic significance; the parameters negatively affecting survival were hemoglobin and age. In conclusion, the cytogenetic profile of SMZL is distinct from other B-cell lymphomas. Complexity of the karyotype, 14q aberrations, and TP53 deletions are poor prognostic indicators and may be considered together with other clinicobiologic parameters to ascertain the prognosis of SMZL.

Unclassifiable lymphoma with features intermediate between diffuse large B-cell lymphoma and Burkitt lymphoma is a new category of B-cell lymphoma appearing in the new World Health Organization Classification of Tumours of Haematopoietic and Lymphoid Tissues. This lymphoma usually shows MYC rearrangements with non-IGH genes in the setting of a complex karyotype possibly involving BCL2 and, less frequently, BCL6 rearrangements. According to the presence of two or three rearrangements, these lymphomas are called double-hit lymphomas or triple-hit lymphomas (THL), respectively. Here we report two cases of THL with MYC, BCL2, and BCL6 rearrangements and t(3;8)(q27;q24) diagnosed in one center in the last two years.

Mantle cell lymphoma (MCL) is a mature B-cell neoplasm with an aggressive behavior, characterized by the t(11;14)(q13;q32). Several secondary genetic abnormalities with a potential role in the oncogenic process have been described. Studies of large MCL series using conventional cytogenetics, and correlating with proliferation and survival, are scarce. We selected 145 MCL cases at diagnosis, displaying an aberrant karyotype, from centers belonging to the Spanish Cooperative Group for Hematological Cytogenetics. Histological subtype, proliferative index and survival data were ascertained. Combined cytogenetic and molecular analyses detected CCND1 translocations in all cases, mostly t(11;14)(q13;q32). Secondary aberrations were present in 58% of patients, the most frequent being deletions of 1p, 13q and 17p, 10p alterations and 3q gains. The most recurrent breakpoints were identified at 1p31-32, 1p21-22, 17p13, and 1p36. Aggressive blastoid/pleomorphic variants displayed a higher karyotypic complexity, a higher frequency of 1p and 17p deletions and 10p alterations, a higher proliferation index and poor survival. Gains of 3q and 13q and 17p13 losses were associated with reduced survival times. Interestingly, gains of 3q and 17p losses added prognostic significance to the morphology in a multivariate analysis. Our findings confirm previous observations indicating that proliferation index, morphology and several secondary genetic alterations (3q gains and 13q and 17p losses) have prognostic value in patients with MCL. Additionally, we observed that 3q gains and 17p losses detected by conventional cytogenetics are proliferation-independent prognostic markers indicating poor outcome., Supported by: subprograma RETICS, Fondo de Investigación Sanitaria, Instituto de Salud Carlos III; Spanish Ministry of Health, Grant numbers: RD07/0020/2004; 2006RET2039; Instituto de Salud Carlos III (ISCIII), Spanish Ministry of Science, Innovation; the Cancer Research Foundation of Salamanca University (Acción Transversal del Cáncer), Spain; Alexander Von Humboldt Foundation.

Fundamento Aunque la enfermedad de Hodgkin (EH) no se considera como una entidad definitoriade sida, en los pacientes infectados por el VIH presenta unas caracteristicas clinicas ybiologicas diferenciadas. Pacientes y metodos Estudio de las caracteristicas clinicopatologicas y analiticas, de la presenciadel virus de Epstein-Barr (VEB) (reaccion en cadena de la polimerasa) y del pronostico en15 pacientes con EH e infeccion por el VIH diagnosticados en un solo centro durante un periodode 10 anos. Resultados Trece enfermos presentaban signos B; en 10 habia afeccion extraganglionar y12 presentaban una EH avanzada. Los tipos histologicos mas frecuentes fueron la celularidadmixta (6) y la deplecion linfocitica (6). La media (DE) de linfocitos CD4 fue 0,10 (0,08) ×109/l. Se evidencio la presencia del VEB en la biopsia ganglionar en 3 de 4 pacientes analizados.Hubo respuesta completa al tratamiento en 7 de 14 pacientes (50%), la mediana desupervivencia global fue de 26 meses y la probabilidad de supervivencia libre de evento a los2 anos fue del 60%. Conclusiones La EH en los pacientes con infeccion por el VIH se presenta en estadios avanzados,subtipos histologicos desfavorables, frecuente afeccion extraganglionar y presencia de signosB. El pronostico es malo, lo cual se debe, sobre todo, a una baja tasa de respuesta al tratamiento.

Chromosomal abnormalities in Ph-negative metaphases from patients with chronic myeloid leukemia (CML) treated with imatinib have been described in some cases. Trisomy 8 is the most frequent, but monosomy 7 has also been described. However, the association of these chromosomal alterations with myelodysplasia has been scarcely reported. We report the appearance of monosomy 7 in Ph-negative cells, associated with severe dysplasia, in two patients with CML treated with imatinib, with a different outcome: one with a transient evolution and the other evolving to acute myeloid leukemia.

The splenic function of patients followed by the Department of General and Digestive Surgery in the Hospital Universitari Germans Trias i Pujol (HUGTiP) from 1985 to 2003 for different degrees of splenic trauma according to the classification of the American Association for the Surgery of Trauma (AAST) 1994 was quantified and related to the treatment received (non surgical, total splenectomy with or without splenosis and splenectomy plus autotransplantation) to detect splenic dysfunction predisposing the development of postsplenectomy sepsis (PSS).43 patients underwent an isotopic study with dynamic splenic gammagraphy and pitted erythrocytes (Normarsky optics) and submembranous vacuoles (transmission electron microscopy) were evaluated.The non surgical group presented normal phagocytic and filtration function with the median speed of splenic enhancement being 3.46 Kcts/s2 (interval: 0.8-6.98). The percentage of pitted erythrocytes was 2% (0-8.8), the number of pits per erythrocyte was 0.03 (0-0.12) and the percentage of erythrocytes with 1, 2, 3 and 4 pits was 1.6%, 0.4%, 0% and 0%, respectively. The percentage of red cells with submembranous vacuoles was 2.55% (0-5.6), the number of vacuoles per red cell was 0.03 (0-0.06) and the percentage of red cells with 1, 2, 3 and 4 vacuoles was 2%, 0.2%, 0% and 0%, respectively. In the operated group, the splenic enhancement speed was 0.08 Kcts/s2 (0-1.75) (p0.0001). The percentage of pitted erythrocytes was 38% (0.2-64) (p0.0001), the number of pits per erythrocyte was 0.86 (0-1.8) (p0.0001) and the percentage of erythrocytes with 1,2,3 and 4 pits was 16.39%, 7.2%, 3.59% and 2.52%, respectively (p0.0001). The percentage of red cells with submembranous vacuoles was 11.2% (1.8-31.9) (p = 0.0006); the number of vacuoles per cell was 0.16 (p = 0.0022) and the percentage of red cells with 1, 2, 3 and 4 vacuoles was 6.51%, 1.73%, 0.4% and 0.2%, respectively (p = 0.0246, 0.0010,0.0001 and 0.0002, respectively).Splenic function of patients with a history of splenic trauma receiving conservative treatment is normal, independently of the degree of the lesion, thereby reinforcing the use of this therapeutic approach to avoid the development of postsplenectomy sepsis. In the patients treated with splenectomy, with or without splenosis, splenic function was absent or very altered being partially conserved in cases treated with splenectomy plus autotransplantation.