Marta García

Adult precursor B-ALL with BCR/ABL gene rearrangements displays a unique immunophenotype based on the pattern of CD10, CD34, CD13 and CD38 expression

The Philadelphia chromosome (Ph+) reflects a balanced reciprocal translocation between the long arms of chromosomes 9 and 22 [t(9;22)(q34;q11.2] involving the BCRand ABL genes. At present, detection of BCR/ABL gene rearrangements is mandatory in precursor-B-ALL patients at diagnosis for prognostic stratification and treatment decision. In spite of the clinical impact, no screening method, displaying a high sensitive and specificity, is available for the identification of BCR/ABL+precursor-B-ALL cases. The aim of the present study was to explore the immunophenotypic characteristics of precursor B-ALL cases displaying BCR/ABL gene rearrangements using multiple stainings analyzed by quantitative flow cytometry in order to rapidly (<1 h) identify unique phenotypes associated with this translocation. From the 82 precursor-B-ALL cases included in the study 12 displayed BCR/ABL gene rearragements, all corresponding to adult patients, four of which also displayed DNA aneuploidy. Our results show that BCR/ABL+ precursor B-ALL cases constantly displayed a homogeneous expression of CD10 and CD34 but low and relatively heterogeneous CD38 expression, together with an aberrant reactivity for CD13. In contrast, this unique phenotype was only detected in three out of 70 BCR/ABL− cases. Therefore, the combined use of staining patterns for CD34, CD38 and CD13 expression within CD10-positive blast cells is highly suggestive of BCR/ABL gene rearrangements in adults with precursor B-ALL.

Safety and efficacy of cyclophosphamide, adriamycin, vincristine, prednisone and rituximab in patients with human immunodeficiency virus‐associated diffuse large B‐cell lymphoma: results of a phase II trial

Immunochemotherapy with cyclophosphamide, adriamycin, vincristine, prednisone and rituximab (R‐CHOP) is the standard treatment in non‐immunosuppressed patients with diffuse large B‐cell lymphoma (DLBCL), but its adequacy has not been definitively established in patients with human immunodeficiency virus (HIV)‐related lymphoma. This phase II trial aimed to evaluate the safety and efficacy of six cycles of R‐CHOP in patients with HIV‐related DLBCL and to determine whether response to highly active antiretroviral therapy (HAART) had prognostic impact. Patients were eligible if they had performance status <3 and absence of active opportunistic infections. Eighty‐one patients were enrolled, 57 in stages III or IV, International Prognostic Index (IPI) 0 or 1 (n = 26), 2 (n = 19), 3 (n = 20) and 4 or 5 (n = 16), and median CD4 lymphocyte count of 0·158 × 109/l. The main adverse events were neutropenia (48% of cycles) and infections (10% of cycles), which were fatal in seven patients. Complete response was achieved in 55 (69%) patients, with an estimated 3‐year disease‐free survival of 77% and 3‐year overall survival of 56%. IPI score and virological response to HAART were the prognostic parameters for response and survival. In HIV‐related DLBCL R‐CHOP is feasible, safe and effective. The prognosis depends on lymphoma‐related parameters and on the response to HAART.

Optical Spectroscopy of Hybrid Sol-Gel Coatings Doped with Noble Metals

Sol-gel coatings in the xM⋅ (100-x) SiO2 system, (M = Cu, Ag and Au) x =0.1–10 mol%), are deposited on soda lime glass slides by using silicon tetramethoxide Si(OCH3)4) and methyltriethoxysilane (SiCH3[OCH2CH3]3) as silica precursors. Anhydrous CuCl, CuCl2 ⋅ 2H2O, Cu(NO3)2 ⋅ 3H2O, CuSO4 ⋅ 5H2O, AgNO3 and HAuCl4 ⋅ 3H2O are used as copper, silver and gold sources. Coatings with thicknesses ranging from 100 to 900 nm are deposited on the subs trates by dip-coating and subsequently densified at 500°C for 1 h in air. Spectroscopic studies of the coatings as a function of the thicknesses and the metal concentration are carried out by photoluminescence (PL) and optical absorption (OA). In addition, direct observations of some gold coatings were performed by transmission electron microscopy (TEM). Results indicate that for silver and copper containing coatings the excitation and emission spectra arise from electronic transitions in Ag+ and Cu+ ions and no significant absorption bands due to colloidal precipitation are observed. Gold containing coatings show purple coloration due to an absorption peaking in the 520–560 nm range, which is characteristic of gold colloids. The presence of these colloids is confirmed by TEM observations.

Diagnosis of secondary myelodysplastic syndromes (MDS) following autologous transplantation should not be based only on morphological criteria used for diagnosis of de novo MDS

Secondary myelodysplastic syndromes (MDS) are increasingly being reported after autologous transplantation. Transient dysplastic changes have also been observed after this type of treatment. However, to the best of our knowledge no systematic morphological analysis has been perfomed to determine the influence of stem cell transplantation on bone marrow morphology. In 53 patients undergoing autologous transplantation, we evaluated the bone marrow, before and 6 and 12 months after the transplant, in order to analyze the appearance of dyshemopoietic changes, assessed according to a pre-established score. We also studied 25 bone marrow samples obtained at the time of diagnosis, prior to treatment, but we did not find morphological atypia. Six months after transplant, cellularity and thrombopoiesis had decreased in 38% and 49% of patients respectively, although 1 year after the process they were normal in most cases. Myelodysplasia was already present in bone marrow before transplantation and continued to be in evidence for a long time afterwards. This suggests that chemotherapy and radiotherapy used prior to transplantation are responsible for dysplastic changes. The myeloid line was the most affected with 100% of patients showing dysgranulopoiesis 1 year after autografting. Cytopenias were observed in 51% and 44% of patients 6 and 12 months after transplantation. Moreover, concomitant presence of cytopenia and myelodysplasia was observed in 37.7% of patients at 6 months after transplantation and 25% at 12 months, and therefore they could be diagnosed with MDS. These data contrast with the incidence of secondary MDS reported in earlier publications. According to these findings, the value of the French–American–British Co-operative Group criteria for the diagnosis of MDS following autologous transplantation is questionable. Moreover, since dyshemopoietic features are almost always present after autologous transplant, morphological criteria are not useful for early recognition of patients with secondary MDS after transplantation.

Long‐term follow‐up of patients with HIV‐related diffuse large B‐cell lymphomas treated in a phase II study with rituximab and CHOP

In the highly active antiretroviral treatment (HAART) era, the addition of rituximab (R) to standard CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone) chemotherapy (Boue et al, 2006; Ribera et al, 2008), CHOP-like regimens (Sparano et al, 2010) or infusional therapies (Spina et al, 2005) has proven to be feasible and effective in Phase II trials and in cohort studies (Wyen et al, 2011) in patients with human immunodeficiency virus (HIV)-related B-cell lymphomas. On the other hand, the incidence of non-acquired immunodeficiency syndrome (AIDS)-related malignancies has increased among patients with HIV infection under HAART (Patel et al, 2008; Crum-Cianflone et al, 2009). However, there is scarce information regarding second malignancies or other AIDS-related events occurring in HIV-infected patients who have responded to treatment for lymphoma. We analysed the long-term follow-up of patients with HIVrelated diffuse large B-cell lymphoma (DLBCL) in complete response (CR) included in a Phase II clinical trial of R-CHOP conducted by the Spanish PETHEMA (Programa para el Estudio y Tratamiento de las Hemopatias Maligna), GESIDA (Grupo de Estudio de SIDA), GELTAMO (Grupo Español de Linfoma y Trasplante de Médula Ósea) and GELCAB (Grupo para el Estudio de los Linfomas Catalano-Balear) groups (Ribera et al, 2008). Eighty-one patients with HIV-related DLBCL were included in the Phase II trial of rituximab and CHOP (R-CHOP) therapy. With a median 2-year follow-up (range: 0·1–5·4) of patients alive at the time of publication, the 5-year overall survival (OS) [95% confidence interval (CI)] probability was 56% (43–69%). In the present study the following events were recorded in the 55 patients in CR: relapse of the lymphoma, opportunistic infections (OI) and other cancers. HIV viral load and CD4 lymphocyte count at the nearest time-point to the event were also recorded. Comparison of these counts between patients with and without events was performed by the Mann–Whitney U-test for CD4 lymphocyte count and the chi-square test for viral load (categorized as negative or positive [>50 copies/ml]. Cumulative probabilities of OI and second cancers, as well as OS and event-free survival (EFS) probabilities were calculated by the Kaplan and Meier method. In patients with more than one event the date of the first event was considered for analyses. This follow-up analysis was performed in October 2011. At the time of the analysis the median follow-up of living patients was 6·2 years (range: 4·6–9·5). One patient in CR was lost to follow-up, eight had presented with lymphoma relapse and five had OI (meningoencephalitis [2], Pneumocystis jiroveci pneumonia [1], varicella pneumonia [1], pneumoccal pneumonia [1], oesophageal candidiasis [1], and cytomegalovirus cholitis [1]. Two patients each had two OI. Five patients developed a second cancer (invasive carcinoma of the cervix [1], squamous lung cancer [1], lung adenocarcinoma [1], pancreatic adenocarcinoma [1], and metastatic sarcoma of unknown origin [1]). One patient presented an OI (varicella pneumonia) followed by pancreatic adenocarcinoma 5 years later. The median time of OI appearance was 0·18 years (range 0·02–6·98) and the cumulative probability of OI at 8 years was 15% (95%CI: 7–23%). For second cancers, these values were 2·77 years (range 2·09–4·71) and 12% (95%CI: 2–22%), respectively. Fifteen patients died: six due to lymphoma relapse, three to OI, four to second cancer and two for other reasons (sudden death in one and violent death in the other). The 8-year OS probability for the cohort of 55 patients in first CR of the lymphoma was 64% (95%CI: 45–83%) (Fig 1A) and the EFS probability was 59% (95%CI: 42–76%) (Fig 1B). The 8-year OS probability for the whole series of 81 patients was 46% (95%CI: 31–61%), representing a 10% reduction in survival with respect to the original publication. No differences in the HIV viral load were observed between patients with and without events (Table I). However, patients with events had a significantly lower CD4 lymphocyte count compared to those without events. As expected, this difference was especially evident for OI. This long-term follow-up study of HIV-infected patients with DLBCL treated with R-CHOP shows a remarkable frequency of OI and second cancers, having a significant impact on the survival probability for this group of patients. Lower CD4 lymphocyte counts were observed in the group of patients with these events than in those patients without these complications. The use of HAART has improved the prognosis of HIVrelated lymphomas, leading to an increase in survival, similar to that of the non-immunosuppressed patients in some matched cohort studies (Navarro et al, 2005), and the International Prognostic Index has proven to be useful for prognosis assessment (Bower et al, 2005). The incidence of non-AIDS-related malignancies has increased in the HAART era (Patel et al, 2008; Crum-Cianflone et al, 2009). However, there is scarce information regarding second cancers occurring in patients that have been successfully treated for lymcorrespondence

Transfusion dependence development and disease evolution in patients with MDS and del(5q) and without transfusion needs at diagnosis

Patients with isolated del(5q) and MDS are considered to have good prognosis as compared to other MDS subtypes. Most patients suffered of anemia and 50% of them required transfusions at diagnosis. It is known that for patients with MDS and del(5q) in transfusion dependence(TD), Lenalidomide is the first choice treatment. However, there are no data regarding natural evolution of anemia in patients diagnosed in MDS and del(5q) without TD, factors that may impact on the development of TD or disease outcome. In the present study we have performed a retrospective multicenter analysis on 83 patients with low-int 1 MDS and del(5q) without TD. During the study 61 patients became TD at a median of 1.7 years and only the Hb level 9 g/dL was associated with poorer TFS (p = 0.007) in the multivariate analysis. Among these 61 TD patients, 49 received treatment (19 Lenalidomide). Median follow up was 48 months, estimated OS at 2 and 5 year was 92% and 50% respectively. In the multivariate analysis for OS, platelets <100,000 mm(-3) and Lenalidomide treatment retained the statistical significant impact. LFS at 2 and 5 years was 86% and 73% respectively, and median time to sAML was 8.16 years (CI 95%: 6.05-10.27). In the multivariate analysis only thrombocytopenia retained statistical significance. In summary, this retrospective study show that level of Hb is an important parameter in order to determine the time until TD, it should be also stressed the importance of an early treatment in order to prevent TD development and shorter survival.