Refractory anemia with ringed sideroblasts associated with thrombocytosis: comparative analysis of marked with non-marked thrombocytosis, and relationship with JAK2 V617F mutational status.

The World Health Organization (WHO) classification of tumors of hematopoietic and lymphoid tissues (2001) defined a provisional entity named refractory anemia with ringed sideroblasts associated to marked thrombocytosis (RARS-MT). Diagnosis of RARS-MT requires more than 15% of ringed sideroblasts in bone marrow aspirate and the existence of a thrombocytosis in blood, with a platelet count above 600 x 10(9)/L. Nevertheless, controversy exists regarding this platelet count "cut-off" value and, when RARS-MT was defined, the JAK2 mutation and its importance in the study of myeloproliferative disorders was unknown. We present the results of a Spanish retrospective multicentric study, which includes 76 cases of RARS with associated thrombocytosis (platelet count above 400 x 10(9)/L) at diagnosis (RARS-T), 36 of them with a platelet count above 600 x 10(9)/L. Our aim was to analyze their clinical, analytical and morphological characteristics, and to establish correlations with the JAK2 mutational status.

Recent improvements in outcome for elderly patients with de novo acute myeloblastic leukemia.

A retrospective analysis was performed on 263 consecutive patients aged over 60 with de novo acute myeloid leukemia (AML) diagnosed in a single institution between 1979 and 1998. Eighty-nine patients (33%) received only palliative treatment, while 174 patients (67%) were treated with different intensive chemotherapy regimens. The 5- and 10-year overall survival (OS) for the whole series was 7.7+/-1.2 and 4.3+/-1.6%, respectively. For patients receiving chemotherapy, OS was 10.5+/-2.5 and 7+/-2.6%, while for those patients receiving supportive treatment it was 1.1+/-1.1 and 0%, respectively (P=0.002). Within the group of patients receiving chemotherapy, the complete remission (CR) rate was 46%; treatment failure rate was 54% (36% due to treatment-related mortality and 18% due to resistant disease). Variables influencing CR rate were FAB subtype, CD7 positivity, chemotherapy regimen, creatinine level, hepatomegaly, and period of diagnosis. Median disease-free survival (DFS) duration was 8.4 months with a probability of being disease-free at 10 years of 10+/-5%. There were no significant differences in DFS according to age. According to the period of diagnosis (1979-1986 vs. 1987-1998), improvements in the CR rate (27 vs. 56%, P=0.0002), and OS (10.9 vs. 15.7 months, P=0.0007) were observed. This large single-center study of unselected de novo AML elderly patients substantiates the progressive improvement achieved in the management of elderly patients with AML, probably due to an improvement in supportive care and the administration of conventional induction chemotherapy.

[Lymphocyte subsets in blood donors]. / Subpoblaciones linfocitarias en donantes de sangre.

OBJECTIVES: To establish the normal values among the different lymphocyte subsets in peripheral blood, measured by surface antigen expression, in healthy population. To observe if there are potential differences in respect of age, gender or sample origin. To compare the absolute lymphocyte number obtained with flow cytometry and with a cellular counter. MATERIAL AND METHODS: Longitudinal and prospective study performed with 100 samples of blood donors. Direct immunofluorescence with triple color staining was made on whole blood, red blood cells were then lysed and samples were analysed with a flow cytometer. The lymphocyte subsets studied were the T lymphocytes and their subsets (CD4 and CD8 lymphocytes), the B lymphocytes and Natural Killer cells population. The absolute lymphocyte count was performed with an automatic cellular counter. Donor data such as age, gender and origin were recorded and were statistically analysed. RESULTS: Normal ranges from the studied parameters are similar to other series. Comparison with gender, origin or age groups gave no significant difference, although there seems to be a tendency to decrease with the ageing of the population. Total absolute lymphocyte number did not differ between the results from the cellular counter or from the flow cytometer. CONCLUSIONS: We found no differences in the absolute lymphocyte number nor in the lymphocyte subsets studied (T lymphocytes, CD4 lymphocytes, CD8 lymphocytes, B lymphocytes and NK cells) with respect to gender, age or sample source. We have established reference ranges for our laboratory. We have not found significant differences in the absolute lymphocyte number measured with cell counter or with flow cytometer.

Acute myeloblastic leukemia with minimal myeloid differentiation (FAB AML-M0): a study of eleven cases.

The main clinical, morphological, cytochemical, immunological features and therapy results of eleven patients diagnosed as acute myeloblastic leukemia M0 (AML-M0) are reported here. There were no clinical characteristics, abnormalities on physical examination or initial laboratory parameters that distinguished these eleven patients. Bone marrow aspirates were hypocellular in four patients. The leukemic cells were undifferentiated by light microscopy and myeloperoxidase (MPO) and/or Sudan Black B (SBB) stains were negative in all cases. Myeloid differentiation antigens were present on the leukemic cells of all eleven patients, whereas B and T cell markers were clearly negative except for CD4 and CD7 antigens. Whatever the treatment employed survival was very short. Eight of the eleven patients were treated and two achieved complete remission (CR) but only one of them is alive in continuous CR. Our results like those previously reported, suggest that AML-M0 patients have a very poor prognosis with standard induction therapies and should perhaps be considered for experimental therapeutic approaches.

[Clinical profile and course of paroxysmal nocturnal hemoglobinuria]. / Perfil clínico y evolutivo de la hemoglobinuria paroxística nocturna.

PURPOSE: To evaluate the clinical characteristics, laboratory findings, course, treatment and survival of a series of patients with PNH. PATIENTS AND METHODS: The clinical records of 21 patients diagnosed of PNH in the La Fe University Hospital between 1970 and July 1991 were revised. Positivity to Ham's and sucrose tests was the major diagnostic criterion. Haemolysis was studied by means of reticulocyte count, unconjugated bilirubin, LDH levels, haptoglobin, serum iron, and haemosiderinuria. The survival analysis was performed in accordance with Kaplan and Meier. RESULTS: The median age in the group was 38 years (range: 18-72 years) and the M/F ratio was 11/10. The commonest symptoms at onset were weakness (76%), dark urine (47%), jaundice (33%), and purpura (24%). All the patients had anaemia, six of them (28%) presenting with aplastic anaemia. The complications most frequently seen included thrombosis (documented in 7 cases, clinically suspected in 6 others), infection (6 cases) and haemorrhage (6 cases). One patient developed aplastic anaemia after 16 years of follow-up, and another one had AML. None of the patients with PNH-associated aplastic anaemia developed thrombosis. The incidence of severe cytopenia was: Hb < 80 g/L, 62%; platelet count < or = 20 x 10(9)/L, 29%, and white cell count < or = 2 x 10(9)/L, 14%. The haemolysis-related findings were as follows: unconjugated hyperbilirubinemia, 100%; haemosiderinuria, 87%; decreased haptoglobin, 80%. Transfusion support consisted of washed red cells (total amount, 1,684 units) and platelets (137 units). A female patient with anticoagulant therapy developed haemolysis after non-isogroup plasma transfusion. Five patients required no transfusions. One patient was subjected to splenectomy and other underwent a successful bone-marrow transplant. The cause of death was related to PNH in three of the five patients who had died when this study was finished (mesenteric thrombosis, subcapsular haematoma of the liver and AML). The actuarial survival at 10 years was 68%. CONCLUSIONS: 1) The incidence of PNH is very low in our experience, excessive delay in diagnosis being the rule, as in other reported series. 2) Early anti-thrombotic treatment should be carried out in PNH whenever severe thrombocytopenia is not associated. 3) Allogeneic BMT is the only curative treatment.

[Culture of cells forming granulomonocytic colonies in bone marrow (GM-CFU) in myelodysplastic syndromes and their relation to hematological findings and FAB subtype]. / Cultivo de células formadoras de colonias granulomonocíticas de medula ósea (CFU-GM) en síndromes mielodisplásicos y su relación con los hallazgos hematológicos y subtipo FAB.

The results of "in vitro" culture of granulomonocytic precursor cells (CFU-GM) of the bone marrow from 44 patients were analysed in the present work. The correlation with the patient's haematological characteristics, their FAB subtypes (i.e., 6 cases of refractory anaemia (RA), 11 of acquired sideroblastic anaemia (ASA), 15 cases of refractory anaemia with excess blasts (RAEB), 5 cases of RAEB in transformation (RAEBT) and 7 cases of chronic myelomonocytic leukaemia (CMML), and the survival were examined as well. The technique used for cell culture was that of Pike and Robinson, following the classification proposed by Florensa for estimating the growth patterns. Anomalies of the myeloid clonal proliferation were found in 81% of the cases. There was direct correlation between the number of aggregates and the polymorphonuclear cell count, whereas the highest number of blast cells coincided with increased number of clusters in cultures. CNNL showed the highest aggregate counts. The B growth pattern (both colony and aggregate growth) was most frequently seen in CMML; pattern C2 (decreased colonies with increased aggregate count) appeared in RAEB, RAEBT and CMML, and pattern C3 (decrease of both colony and aggregate counts) was found only in RA and ASA. None of the culture findings appreciably associated with the survival.

11q23 abnormalities in children with acute nonlymphocytic leukemia (M4-M5). Association with previous chemotherapy.

Cytogenetic studies of 12 patients aged less than 14 years with acute nonlymphoblastic leukemia (ANLL) (M4-M5) showed structural abnormalities on chromosome 11 at band q23-q24 in five cases (41.8%). Four of these 12 patients had ANLL (M4-M5) after treatment with cytostatics for non-Hodgkin lymphoma in one case and for an acute lymphoblastic leukemia (ALL) in the other three. Three of these four cases had 11q23 abnormalities [one [one 46,XY,t(11;17)(q23;25); another 47,XY,+8,-15,del(11)(q23),+der(15)t(15;?)(p11;?); the third 47,XX,+8,t(3;17) (p11;q25),t(4;11)(q21;q23)] and one had a normal karyotype on being diagnosed ANLL (M4-M5). The notable increase of ANLL (M4-M5) in our patients who had received cytostatics as treatment for a previous neoplasia makes evaluation of our results timely in comparison with those of other groups who use these therapeutic protocols.

Acute Promyelocytic Leukaemia. Relationship of Cytochemical Pattern with Clinical, Morphologic and Immunological Characteristics, and Outcome.

In a series of 61 acute promyelocytic leukemia (APL) cases multiple clinical and biological characteristics were studied in order to assess their prognostic significance and to evaluate the relationship of cytochemical pattern with clinical, morphologic, and immunological characteristics and outcome. Alpha-naphthyl acetate esterase (ANAE) activi1:y was expressed in 23 out of 41 cases (56%). Sodium fluoride-sensitive naphthol AS-D acetate esterase was evident in only one out of 22 cases (5 %). ANAE reactivity was more frequently found in typical hypergranular APL (P = 0.003) and was associated with a shorter interval symptoms-diagnosis (P = 0.04). The expression of HLA-DK, CD33, CD13 and CD11b did not show any correlation with ANAE reactivity. Complete remission (CR) was achieved in 64 % of treated patients. The actuarial median remission duration was 22 ± 3 months. CR rate was significantly related to the presence of fever, age, absolute reticulocyte count, and albumin level, while the remission duration was associated with absolute reticulocyte, leucocyte and granulocyte counts, and albumin level. In our study ANAE activity did not reflect any important clinical and biological features of APL, apart from its obvious correlation with morphological subtype. In addition ANAE activity did not correlate with early response to therapy nor related with long-term prognosis.

[Hematologic alterations and reduction in serum folate in epileptics treated with anticonvulsants]. / Alteraciones hematológicas y descenso de folatos séricos en epilépticos tratados con anticonvulsivos.

A prospective study of the incidence of toxic hematologic effects of antiepileptic drugs was carried out in a series of 104 epileptic patients treated with phenytoin alone or combined with other drugs, and in 30 patients treated with other anticonvulsants. A slight decrease in hemoglobin values and a slight increase in mean corpuscular volume and mean corpuscular hemoglobin was observed in both groups. These changes are typical of the hyperchromatic megaloblastic anemias, although a statistically significant relationship between folates and hemoglobin could not be established. Moreover, we found leukopenia and lymphopenia in the group treated with hydantoins, and moderate eosinophilia in both groups. Changes in metabolism of vitamin B12 and folate were frequent, with decreased values in more than 30% of patients. A significant inverse correlation was observed between serum levels of folates and phenytoin, which suggests a direct toxic effect of the drug. No major dyscrasias were noticed.