Inositide-specific phospholipase c beta1 gene deletion in the progression of myelodysplastic syndrome to acute myeloid leukemia.

Myelodysplastic syndrome (MDS) is an adult hematological disease that evolves into acute myeloid leukemia (AML) in about 30% of the cases. The availability of a highly specific probe moved us to perform in patients affected with MDS/AML, associated with normal karyotype, painting and fluorescence in situ hybridization (FISH) analysis aimed to check the inositide-specific phospholipase C (PI-PLC) beta1 gene, a player in the control of some checkpoints of the cell cycle. Here we present a preliminary observation in which FISH analysis disclosed in a small group of MDS/AML patients with normal karyotype the monoallelic deletion of the PI-PLCbeta1 gene. On the contrary, PI-PLC beta4, another gene coding for a signaling molecule, located on 20p12.3 at a distance as far as less than 1Mb from PI-PLCbeta1, is unaffected in MDS patients with the deletion of PI-PLC beta1 gene, hinting at an interstitial deletion. The MDS patients, bearing the deletion, rapidly evolved to AML. The data suggest the possible involvement of PI-PLCbeta1 in the progression of the disease and pave the way for a larger investigation aimed at identifying a possible high-risk group among MDS patients with a normal karyotype.

Spectral karyotyping (SKY) refinement of a complex karyotype with t(20;21) in a Ph-positive CML patient submitted to peripheral blood stem cell transplantation.

A patient with a Ph-positive chronic myeloid leukaemia (CML) was submitted to allogeneic peripheral blood stem cell transplantation from an HLA-haploidentical related donor 7 years after the diagnosis. Six months later, he showed a disease relapse while cytogenetic analysis displayed a complex karyotype. To characterise the chromosomal rearrangements spectral karyotype (SKY) analysis was used. This redefined all chromosome rearrangements and revealed a t(20;21)(q11;q22). FISH analysis with a specific probe for the AML1 gene disclosed disruption of this gene which was partially translocated on to the long arm of chromosome 20. It is likely that this rearrangement, unusual for CML, was implicated in the disease evolution towards blastic crisis (BC).

p53 loss and point mutations are associated with suppression of apoptosis and progression of CML into myeloid blastic crisis.

A longitudinal investigation using fluorescence in situ hybridization (FISH) analysis, PCR-SSCP, and in situ detection of apoptosis by the terminal deoxynucleotidyl Transferase (TdT) method was carried out on 13 chronic myelogenous leukemia (CML) patients to study the p53 gene behavior and the apoptotic process during the course of the disease. At diagnosis, FISH showed no loss of the p53 gene on interphase nuclei, and no point mutation was detected by polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) and sequencing. During the disease course, FISH analysis showed a significative loss of allele (LOA) rate for the p53 gene in eight patients that in seven cases was associated with a suppression of apoptotic process and the progressive expansion of the p53+/p53- clone. DNA sequencing showed in two of these eight patients a point mutation on the other allele, consisting in the formation of a stop codon in one case, and in a frameshift mutation in the other. Six patients had a myeloid blastic crisis (BC), five a lymphoid BC, and the other two an erythroid and an undifferentiated BC, respectively. All patients with myeloid BC and the one with undifferentiated BC disclosed a progressive expansion of the clone with p53 loss that was associated with a significant reduction in apoptosis. On the contrary in the 5 patients with lymphoid BC no significant p53 LOA rate was observed during the course of the disease. In these patients apoptotic process also persisted in the acute phase although in a lower rate as compared to CP.

Leukemic evolution in three patients with myelodysplastic syndrome and unusual chromosome changes.

The authors describe three patients with myelodysplastic syndrome without a history of exposure to chemical agents and who showed many chromosome rearrangements not previously reported in this hematologic disorder, and a rapid outcome of the disease. The authors discuss the significance of the chromosome changes, suggesting, in agreement with others, that patients with complex rearrangements have a poor prognosis.

Complex translocations of the Ph chromosome and Ph negative CML arise from similar mechanisms, as evidenced by FISH analysis.

The authors report on 13 patients with chronic myeloid leukemia (CML) studied by serial karyotyping and fluorescence in situ hybridization (FISH) of their bone marrow cells. Ten patients had complex translocations of the Ph chromosome while the remaining three were Ph negative. FISH analysis revealed in all 13 patients the translocation of the ABL protooncogene into chromosome 22 at band q11. Moreover, in all complex translocations but one, FISH with a chromosome 22 painting probe demonstrated on one chromosome 9 at band q34 the presence of material from chromosome 22, in addition to signals on the third chromosome involved in complex changes. Therefore, in this study complex translocations appeared as secondary changes resulting from two consecutive translocations with a total of at least four breaks. The first translocation gave rise to the standard t(9;22)(q34;q11). The second one included a break distal to the original breakpoint at band 9q34 and another one on a third chromosome. Furthermore FISH using S1 and S15 probes, mapped at band 22q11.2 or 22q12, gave evidence that in complex translocations the secondary breakpoint on der(9) was in the translocated segment 22q11-qter between bands q11 and q12. FISH analysis also disclosed the presence of material from chromosome 22 on one chromosome 9 in the three patients with Ph negative CML, demonstrating that in these cases a retranslocation between chromosomes 9q+ and 22q- had occurred. Consequently, the four-break mechanism could also be invoked for the three Ph negative CML patients.

Cytogenetic survey of 80 patients with acute nonlymphocytic leukemia.

The authors report a cytogenetic survey of 80 patients with acute nonlymphocytic leukemia. The prognostic value of chromosome aberrations has been evaluated with three methods. The first one showed that patients with NN or AN bone marrow cellularity have a significantly better prognosis than those with AA cellularity; the second method confirmed the relatively good prognosis for patients with t(8;21) and abnormal 16 and a poor one for those with rearrangements of chromosomes 5 and/or 7. The authors also noted, surprisingly, that patients with hyperdiploidy had a significantly poorer prognosis than those with hypodiploidy and especially pseudodiploidy. The third method showed that patients with very complex karyotypes and a worse outcome than those with simple changes. Finally, they discuss the prognostic value of unusual and/or undeciphered chromosome changes detected in 18 patients, with a mean survival of 9.6 months, showing that these changes have a negative prognostic significance.

Chromosome changes in 19 patients with Waldenström's macroglobulinemia.

We report on 19 patients with Waldenström's macroglobulinemia (WM) who were studied cytogenetically at the onset and during progression of the disease. We found a high frequency of chromosome changes confirming the claim of other authors that, during progression of the disease, a large number of residual neoplastic cells, insensitive to conventional chemotherapy, persist. In turn, this may be the cause of the difficulty of inducing remission (21% of cases) and of the short survival (mean, 35 months). In our experience it is difficult to identify the primary chromosome abnormalities because of the late clinical stage at which the chromosomes were examined. However, changes involving chromosomes #10, #11, and #12 may be unfavorable events in patients with WM.

Autologous blood stem cell transplantation in malignant lymphomas: an Italian Cooperative Study.

Twenty-three patients with malignant lymphoma, (7 Hodgkin's, and 16 non-Hodgkin's) in different phases of disease were autografted in 4 Italian Haematology institutions using only chemotherapy-mobilized blood stem cells (BSC) collected by apheresis. Clinical and laboratory data were analysed centrally and showed mean collection yields of 8.1 x 10(8) kg mononuclear cells (MNC) (SE 0.5; range 2.6-13.8) and 24.1 x 10(4) kg CFU-GM (SE 7.4; range 1.4-162.9). The mean times required to attain 0.5 x 10(9)/l neutrophils and 50 x 10(9)/l platelets after marrow-ablative high-dose chemo+radiotherapy and BSC reinfusion were 14.9 days (SE 1.5; range 7-38) and 18.6 days (SE 2.6; range 6-49) respectively. The incidence of early deaths was < 5% and the requirement for support with blood product transfusion was moderate. The progression free survival (PFS) is > 50% at 3 years with a median follow-up of 17.3 months. Results were significantly better for patients autografted in remission. These results suggest that autologous blood stem cell transplantation (ABSCT) may be proposed for the primary treatment of poor prognosis malignant lymphomas. However, ABSCT needs to be compared with autologous bone marrow transplantation (ABMT) followed by infusion of growth factors to accelerate recovery.

Impact of a new dosing regimen of epoetin alfa on quality of life and anemia in patients with low-risk myelodysplastic syndrome.

This study evaluated the impact of a new epoetin alfa dosing regimen on quality of life (QOL), transfusion requirements, and hemoglobin (Hb) levels in 133 patients with low-risk myelodysplastic syndrome (MDS) and Hb < or =10 g/dl. Epoetin alfa 40,000 IU was given subcutaneously twice weekly; after 4 weeks, the dose could be reduced to 40,000 IU weekly in patients achieving erythroid response. QOL was assessed using the functional assessment of cancer therapy-anemia (FACT-An) questionnaire. FACT-An scores increased on average by 7.5 after 4 weeks and by 8.8 after 8 weeks compared with baseline. FACT-An scores were positively associated with Hb values (r=0.53, P<0.01). The mean FACT-An score increase at week 8 was 10.2 in responders and 5.6 in nonresponders. The overall erythroid response rate at week 8 was 68%: 74% in transfusion-independent patients and 59% in transfusion-dependent patients. Of all responders at week 8, response was maintained in 86% at week 12, 71% at week 16, 65% at week 20, and 54% at week 24. Treatment was generally well tolerated. Our data provide new and encouraging results regarding the benefits of 40,000 IU biweekly induction doses followed by 40,000 IU weekly in improving QOL, correcting anemia, and reducing transfusion requirements in low-risk MDS patients.

Cytogenetic survey of sixty-one patients with preleukemic syndrome including myeloproliferative and myelodysplastic diseases.

The authors report on a cytogenetic survey of 61 patients with preleukemic syndrome (PLS). Of these, 41 had a myeloproliferative disease (MPD) and 20 a myelodysplastic syndrome (MDS). Clonal chromosome abnormalities appeared in 24 patients (39.3%) at disease onset. Such changes had a frequency of 26.8% in patients with MPD and 65% in those with MDS. The authors stress the usefulness of ethidium bromide high resolution techniques. They allow obtaining a larger number of metaphases and elongated chromosomes with higher banding resolution and could account for the frequent detection of chromosome changes in most groups of MDS patients in the present series. Moreover, they discuss the possible significance of some chromosome aberrations suggesting that patients with MPD may live longer than those with MDS because of their higher frequency of normal karyotypes.

Autologous blood stem cell transplantation as intensive consolidation therapy in hematological malignancies.

Ten consecutive patients (pts), suffering from hematological malignancies (5 NHL, 3 ANLL and 2 HD), received high doses of radio-chemotherapy and autologous peripheral blood stem cell (PBSC) transplantation. Seven of them were in 1st CR, two in 3rd CR and one in refractory status. The recruitment of PBSC was performed during the conventional non-intensive schedule of therapy. The median interval between the remission and the transplant was 6 months (3-15). The pts received a median of 0.96 x10(9) MNC/Kg b.w. and of 3.8 x10(4) CFU-GM/Kg b.w. Hemopoietic recovery occurred promptly and the median number of days to reach WBC greater than 1.0 x10(9)/L was 10 (7-15), PMN greater than 0.5 x10(9)/L was 12 (10-34), platelets greater than 50 x10(9)/L was 13 (10-41) and reticulocytes greater than 20 x10(9)/L was 12 (10-15). Three pts relapsed after 14, 12 and 6 months, respectively. Two of them achieved a further remission and one died. The remaining seven, are alive and disease-free with a follow-up of 11 months (range 2-27).

Randomized comparison of ceftriaxone versus ceftriaxone plus amikacin for the empirical treatment of infections in patients with altered host defense: microbiological and clinical evaluation.

Two hundred and eighty-four febrile episodes in immunocompromised patients were treated with ceftriaxone alone or in combination with amikacin. In the ceftriaxone-treated group, 60 out of 143 febrile episodes were microbiologically documented, while in the group receiving the combination therapy, there were 32 out of 140 (p = 0.0007). Gram-positive microorganisms were more common than gram-negative ones, accounting for 59 of the 101 isolated bacteria. The ceftriaxone regimen appeared to have a response rate comparable to the combination regimen (73.91 vs. 78.88%). Superinfections occurred under both regimens.

Cytogenetics and acute non lymphocytic leukemia.

The authors report haematologic and cytogenetic data from 47 patients with ANLL, demonstrating the usefulness of cytogenetic studies for the classification as well as for the prognosis of this disorder. Chromosome studies also permitted the classification of marrow cellularity in: all diploid metaphases (NN), diploid and aneuploid metaphases (AN), and all aneuploid metaphases (AA). The remission rate for patients in whom only normal metaphases were detected (NN patients) was 83% while the remission rates were 67% and 33% respectively for patients in whom both normal and abnormal metaphases were seen (AN patients) and for those in whom only abnormal metaphases were noted (AA patients). In all FAB subgroups, complete remission was related to chromosomal abnormalities, except for M4 patients who evidenced a large number of complete remissions, although presenting more chromosomal abnormalities. The longer survival in this subgroup may be related to rearrangements of chromosome 16, which is associated with a better prognosis.