Fluorescence in situ hybridization study of TEL/AML1 fusion and other abnormalities involving TEL and AML1 genes. Correlation with cytogenetic findings and prognostic value in children with acute lymphocytic leukemia.

BACKGROUND AND OBJECTIVES: The TEL/AML1 fusion is the most common genetic abnormality found in childhood acute lymphoblastic leukemias (ALL). Although it is very difficult to identify by conventional cytogenetic techniques it can be readily detected using fluorescence in situ hybridization (FISH). We carried out cytogenetic and FISH studies on 42 children with ALL in order to know the frequency of this translocation in our population, the incidence of TEL and/or AML1 gene alterations, and their correlation with clinical evolution and prognosis. In addition, we performed reverse transcription polymerase chain reaction (RT-PCR) in some cases, confirming the feasibility of FISH techniques in the detection of this translocation. DESIGN AND METHODS: Bone marrow samples were obtained from 42 childhood ALL patients. The copy number of AML1 and TEL genes were studied using fluorescent in situ hybridization with a dual color DNA probe specific for the AML1 and TEL genes. RESULTS: We found a frequency of TEL/AML1 fusion of 17% in our sample. Double TEL/AML1 fusion, lack of TEL signal and extra AML1 signals were frequent additional FISH abnormalities. Duplication of a chromosomal complement, deletion of chromosome 12p arm, and polysomies of chromosome 21 are plausible explanations for these additional FISH findings. However, a relatively high proportion of our cases (9.5%) presented specific amplification of AML1. A statistically significant difference in prognosis was found between patients with and without these additional AML1 or TEL FISH alterations (p<0.02), which could be related to the presence of specific karyotypes. INTERPRETATIONS AND CONCLUSIONS: The frequency of TEL/AML1 fusion is similar to that found in other populations (17%). We found that FISH analysis of AML and TEL is related to the evolution of the disease. The absence of alterations in these genes revealed by FISH could be indicative of bad prognosis, while the presence of alterations is related to a good evolution. Our results suggest that interphase FISH analysis to search for alterations in AML and TEL genes could be extremely useful for complementing cytogenetic studies and for providing additional information about the possible outcome of the disease in patients with ALL.

Flow cytometry and the study of central nervous disease in patients with acute leukaemia.

Central nervous system (CNS) leukaemia is still a matter of debate and new technologies are required to improve the classic morphological definition. One hundred and sixty-eight cerebrospinal fluid (CSF) samples from 31 patients with acute leukaemia were analysed by flow cytometry and conventional cytology. Concordant positive and negative findings were found in 158 samples but 10 produced discrepant results. Cytology seemed to offer more precise information in one CSF sample and flow cytometric accuracy could be demonstrated in five samples. We conclude that flow cytometry is of great help in confirming CNS leukaemia and eliminating other conditions. Therefore, leukaemic patients can benefit from double cytological and flow cytometric CSF studies.

Chronic recurrent multifocal osteomyelitis after acute lymphoblastic leukaemia.

We describe an 8 year old girl who developed chronic recurrent multifocal osteomyelitis (CRMO) in the ilium and clavicle. Treatment for an acute lymphoblastic leukaemia had been finished two months before. After antibiotic therapy, the clinical symptoms improved and no fresh lesions appeared. The aetiology of CRMO is unknown, but we feel that infection may precipitate an immunological reaction.

Infection caused by the nonfermentative gram-negative bacillus CDC group IV c-2: case report and literature review.

A 10-year-old girl with acute lymphocytic leukemia developed nosocomial septicemia caused by the gram-negative bacterium CDC group IV c-2. Recovery of the patient followed appropriate treatment with ceftriaxone, to which the organism was susceptible in vitro. Four other reported cases of infection caused by this organism are reviewed.

[Capnocytophaga spp. bacteremia in compromised patients]. / Bacteriemia por Capnocytophaga spp. en pacientes comprometidos.

BACKGROUND: To analyze the clinical characteristics of three patients with Capnocytophaga spp. bacteremia. METHODS: We have review the clinical charts of three patients with Capnocytophaga spp. bacteremia, two patients with acute leukemia and one epileptic patient with chronic alcoholism. RESULTS: All the patients had oral cavity troubles and were treated with different antimicrobial agents. The three patients recovered, one of them requiring changing of the antimicrobial therapy in one of them. CONCLUSIONS: We should be aware of the importance of Capnocytophaga spp. especially in immunosuppressed patients or in other patients with poor dental hygiene and fever.

[Acute lymphoblastic leukemias off therapy: course of 80 cases after treatment cessation]. / Leucemias agudas linfoblásticas con terapéutica suprimida: evolución postsupresión de 80 casos.

PURPOSE: To retrospectively analyse all the patients diagnosed and treated in the same hospital for acute lymphoblastic leukaemia (ALL) in whom therapy was suppressed after sustained complete remission (CR) for a variable period. PATIENTS AND METHODS: Eighty cases of ALL treated at the Jiménez Díaz Foundation between 1968 and 1991 were revised. Treatment had been suppressed after 60 months of maintained CR (1968-1974) or after 28 months (1986-1991), with a median follow-up after suppression of 6 years. All the patients had been treated with several protocols (FJD-68, FJD/BFM, APO, BFM-83, BFM-86 and BFM-90). Maintained CR, relapses and course after therapy cessation were analysed. Actuarial curves of RC duration and survival were drawn according to the Cutler and Ederer life tables. RESULTS: The age at diagnosis ranged from 1.5 to 68 years. Childhood (< 15 years) ALL presented in 49 cases; 25 were young adults (15-30 years), and 6 others were over 30 years of age. Two of the 80 patients (2.5%) died in CR (astrocytoma and demyelinating leukoencephalopathy). Fifty-eight patients (72.5%) are living in maintained CR; 20 others relapsed (25%), 14 of them (70%) attaining a second CR. Allogenic BMT was carried out in 4 such cases. Four patients are out of any treatment after the second CR. The actuarial curves of CR duration after suppression of therapy gave a stable plateau at 73% for children, at 75% for young adults and at 22% for patients over 30 years. The survival after suppression of therapy showed a plateau at 75% for children and at 86% for young adults. CONCLUSIONS: 1) Relapse was seen in 25% of the patients after cessation of therapy. 2) Relapse occurred in the first two years after therapy cessation in 75% of the instances. 3) At this writing, 78.7% of the patients keep up the CR. 4) Adults over 30 years of age comprise the poorest prognosis. 5) The course of young adults in this series after cessation of therapy is as good as that of children.

Immunophenotypic and Gene Rearrangement Analysis in Null- or T-Cell Neoplasias: Study of 16 Cases.

Immunophenotypic and molecular studies were performed in sixteen cases of T-cell lymphoproliferative disorders. These included eleven patients with peripheral T-cell lymphoma, two thymic lymphomas and three patients with T-gamma lymphocytosis. Peripheral T-cell lymphomas were of both low and high grades. There was one case of Sezary's syndrome, two of small T-cell pleomorphic type, two medium sized T-cell pleomorphic lymphomas, two large T-cell pleomorphic type and four large cell anaplastic T cell lymphomas with activated T cell markers. Two patients had lymphoblastic lymphoma of thymic origin. In this report we attempted to correlate immunophenotypic and molecular characteristics. Rearrangements of the T-cell receptor (TCR) genes were observed in all cases, including those lacking any immunophenotypic markers, and unusual rearrangements of both the TCR and Ig genes were evident in thymic and peripheral T-cell lymphomas. In the cases of T-gamma lymphocytosis, a lymphoproliferative disorder that is not always clearly defined monoclonality was seen in one. The use of the genotypic approach for refining the characterization and diagnosis of some T-cell neoplasias is emphasized. The problems and pitfalls arising from the application of these methods are also discussed.

[Acute promyelocytic leukemias: clinico-biological aspects, prognostic factors, therapeutic response, and possibilities of cure in 34 cases (1970-1988)]. / Leucemias agudas promielocíticas: aspectos clínico-biológicos, factores pronósticos, respuesta terapéutica y posibilidades de curación de 34 casos (1970-1988).

Thirty-four new cases of acute promyelocytic leukaemia (M3) were diagnosed at the authors' Centre between 1970 and 1988 (19 males and 15 females) with ages between 5 and 73 years (median age, 32 years). Three cases were of the hypogranular variant or M3-v (8.8%). The clinical picture included: haemorrhagic diathesis (85%), pallor/malaise (82%), fever/infection (41%), hepatomegaly (26%), splenomegaly (12%). Leucopenia of less than 5 x 10(9)/L was present in 23/34 cases, laboratory signs of DIC in 26/31, increased LDH, over 400 U/mL, in 6/31, and abnormal karyotype in 7/15. One of the patients rejected any treatment; two others died of brain haemorrhage before therapy was started, and seven died in the first two weeks of treatment. Of the 31 patients treated, complete remission (CR) was achieved in 21 cases (67.7%). Allogeneic BMT was carried out in two of them, with further relapse and death. Post-remission treatment was given to the remaining 19 patients, and there were 13 relapses. Six patients have been in CR, 5 of them after cessation of therapy, for the last 1.5-11.5 years. Age under 50 years and leucocyte count below 5 x 10(9)/L at diagnosis were favourable prognostic factors according to the univariate statistical analysis performed. The survival plateau of the actuarial curve was reached beyond 2.75 years by 15% of all the patients treated (33 cases), 23% of the patients who achieved CR (21 cases), 31% of the patients under 50 years of age and 5 x 10(9)/L leucocyte count at diagnosis (15 cases) and 36% of these last achieving CR (13 cases).

[Open lung biopsy in immunocompromised patients with diffuse pulmonary infiltrates]. / La biopsia pulmonar abierta en pacientes inmunocomprometidos con infiltrados pulmonares difusos.

In order to evaluate the diagnostic usefulness of open lung biopsy and to determine how could it influence the treatment and the evolution of the disease, the clinical histories of 19 immunocompromised patients with diffuse lung infiltrates were reviewed. One or more specific diagnosis were obtained in 14 patients (73%) by open lung biopsy. However, the initial treatment was modified, in view of the results of the biopsy only in 3 cases (15%). Only 5 patients survived and were discharged. There were no differences in the survival rate of patients with a specific or a nonspecific diagnosis (11 out of 14 deaths and 3 out of 5 respectively). 5 patients suffered severe complications from the surgical procedure. Open lung biopsy should not be used routinarily in the study of diffuse lung infiltrates in immunocompromised patients.