Cytochemical study of acute promyelocytic leukaemia.

Five cases of acute promyelocytic leukaemia (A.P.L.) are investigated for peroxidase, PAS, toluidine blue, astra blue, alpha-naphthyl esterase, double esterase incubation (naphthol-AS plus naphthol-AS-D chloroesterase), and cellular lysozyme activity. These cytochemical investigations may contribute further characterization of the morphologic type.     

Cytochemical studies in the blastic transformation of chronic granulocytic leukaemia

The cytochemical features of blast cells were studied in 45 patients with blastic phase of chronic granulocytic leukaemia. Various degrees of Sudan black B positivity was characteristic of myeloblastic transformation (23 patients), while in the medullary blast cells of nine patients with myelomonocytic transformation the alpha-naphthyl-acetate esterase showed intensive activity. In two cases the demonstrability of beta-thromboglobulin and factor VIII-related antigen in blast cells showing otherwise PAS, acid phosphatase and alpha-naphthyl-acetate esterase activity referred to megakaryocytic transformation. In six patients with lymphoid blast crisis proliferation of the Sudan negative blast cells with different granular PAS, acid phosphatase and/or beta-glucuronidase positivity was demonstrated. In five cases the cytochemical findings of leukaemic cells indicated biphenotypic and mixed transformation, respectively.     

Bone marrow stromal culture from patients with myelodysplastic syndromes and patients at different stages of acute nonlymphocytic leukaemia

The haematopoietic microenvironment or stroma plays a decisive role for the proliferation and differentiation of haemopoietic cells. We studied if bone marrow cells from patients with myelodysplastic syndromes (MDS) and acute nonlymphocytic leukaemias (ANLL) are altered in their ability to form adherent stromal layer with active haemopoiesis in the Dexter liquid culture. Bone marrow cells were obtained from 24 normal volunteers, 28 patients with ANLL in different stages of the disease and 9 patients with MDS. There are no differences between the stromal layers of patients with ANLL in complete remission and those of normal volunteers after two weeks of cultivation. However, bone marrow cells from patients with ANLL before treatment and from patients in relapse formed a poor adherent stromal layer in most cases. In 6 of 9 cases we found the normal stromal grade of bone marrow cells from patients with MDS. There were qualitative differences in the nonadherent cell population between normal and ANLL patients in complete remission. In most cases we found morphologically recognizable erythroid cells after two-weeks Dexter liquid culture of bone marrow cells from patients with ANLL in complete remission, which were not seen with normal volunteers. This could be an indication of harmful effects on the balance of haematopoiesis caused by previous infiltration with leukaemic cells or/and high-dose chemotherapy.     

Immunotherapy with bestatin for acute nonlymphocytic leukemia in adults

Summary In a cooperative randomized control study of immunotherapy with bestatin in combination with chemotherapy in adults with acute nonlymphocytic leukemia (ANLL), 101 patients (48 in the bestatin group and 53 in the control group) out of 115 patients registered were evaluated as eligible. The bestatin group achieved a statistically significant prolongation of survival compared with the control group in overall ANLL and acute myelogenous leukemia. In the analysis of patient age, the bestatin group achieved a statistically significant prolongation of both the remission duration and survival in patients aged 50 to 65 years, while the differences were not significant in the 15 to 49 age group. The bestatin group tended to achieve a higher rate of reinduction of remission in patients who had recurrence of leukemia. Side effects developed in only 5 (9.6%) of 52 patients treated with bestatin. None of these side effects were particularly serious in nature. It is concluded that bestatin is useful for prolongation of survival of adult patients with ANLL, making for a longer remission duration especially in elderly patients and with few side effects.     

Prognostic significance of cell surface phenotype in adult acute lymphoblastic leukaemia

Summary Forty-two adults with acute lymphoblastic leukaemia were subgrouped according to the membrane marker characteristics of their blast cells. All patients received the same treatment at St. Bartholomew's Hospital. The results indicate that the group expressing the common ALL antigen have a superior remission duration to the ALL antigen-negative, non-T, non-B ALL group and the T-ALL group. No correlation was evident between the first two subgroups and the clinical features of disease.     

Infiltration of central nervous system in adult acute myeloid leukaemia.

Out of 64 consecutive unselected patients with acute myeloid leukaemia studied during 1973-6, five developed clinical evidence of spread to the central nervous system (CNS). Neuroradiological examination showed cerebral deposits in three, in whom rapid symptomatic relief was obtained with radiotherapy. In two of these patients who developed solid intracranial deposits haematological remission could be reinduced or maintained; they were still alive 86 and 134 weeks later. When patients presented with spread to the CNS complicating generalised uncontrolled leukaemia they had short survivals. CNS infiltration may respond dramatically to appropriate treatment provided that it is not associated with generalised uncontrolled leukaemia, which has a poor prognosis. In view of this, routine "prophylaxis" of the CNS in adult acute myeloid leukaemia does not seem justified at present.     

Cytochemical index of the function of the afferent fiber

A correlation between the spike activity and ferrous deposits in the Ranvier nodes of afferent fibers has been revealed in the frog bladder. The degenerated fibers reveal neither deposits, nor spike activity. The native fibers reveal deposits in the presence of spike activity. When the spike activity is suppressed by procainamide the node affinity to ferrous ions is retained. It is suggested that positive cytochemical reactions in the nodes demonstrate the retention of the initial morpho-functional structure of afferent fibers.     

A search for novel tumour suppressor genes for adult acute leukaemia by allelotyping at sub-telomeric chromosomal regions

A search was initiated towards the localization of novel mutated tumour suppressor genes that may be involved in adult leukaemia. For this purpose, we measured the occurrence of loss of heterozygosity (LOH) in nine patients with acute B-lineage leukaemia (ALL) and one with undifferentiated leukaemia (AUL). Eight leukaemias exhibited a diploid karyotype. For each patient, PCR products of 130 polymorphic microsatellite markers, located in subtelomeric areas of every autosomal chromosome arm were analysed to visualize LOH events resulting from reduplication of a single mutated chromosome or from mitotic recombination. These kinds of LOH events contribute most to LOH in model systems but cannot be detected by classical cytogenetic techniques. By comparing allelic PCR products in tumour cells with those in normal cells, LOH was found in tumour cells of one ALL patient at 9p which harbours the known p16^INK44 tumour suppressor gene. In the AUL patient, however, LOH was detected at the telomeres of 4q and 21q, suggesting that these sites may contain novel tumour suppressor genes specifically involved in this form of leukaemia. In the DNA of tumour cells from eight out of 10 patients no LOH was detected. This is in contrast with the general assumption that LOH is a frequent phenomenon in ALL. However, some markers at telomeric regions of chromosomes were already homozygous in the control T-cells of several patients. For instance, we found in the DNA of control cells from one patient five consecutive microsatellites on 9p up to 9p43 which were homozygous and in three other patients homozygosity was observed in band 8q24, which includes the MYC gene. These observations indicate that LOH events already are present in non-cancerous putative stem cells and that mitotic recombination may be a very early event in leukaemogenesis.     

Remission induction and remission maintenance in adult acute nonlymphocytic leukemia employing a modified cytostatic (COAP) regimen.

Thirty adult patients suffering from acute nonlymphocytic leukemia (ANLL) were treated according to a modified COAP regimen. Vincristine, cyclophosphamide, and prednisone were given by push injection, while cytosine arabinoside was infused over periods of 8 h. Nineteen patients (63%) achieved complete remission. Remission maintenance therapy consisted of 6-mercaptopurine daily and methotrexate twice weekly. Later in the study, COAP consolidation and reinduction was added, which improved the median duration of complete remission from 7 to 24 months. Comparison of the results with the literature shows that the modified COAP regimen is one of the most effective treatment schedules for adult ANLL.     

Short-term treatment for acute myelogenous leukaemia.

Short-term treatment with doxorubicin, cytarabine, and 6-thioguanine was given to 91 consecutive adults with acute myelogenous leukaemia. Fifty patients received high doses (regimen I) and 41 very high doses (regimen II). Where possible, six treatment cycles were given (total dose of doxorubicin 450 mg/m2) regardless of the number of cycles required to achieve complete remission. No additional treatment was given. The remission rate was significantly higher with regimen I than with regimen II (34/50 compared with 15/41, p less than 0.01), the latter, more intensive regimen being associated with a greater incidence of fatal infection (13/41 compared with 5/50, p less than 0.01). Duration of remission was, however, significantly longer with regimen II (p less than 0.05); the median has not yet been reached after a minimum follow-up of two years. Intensive short-term treatment is a feasible strategy for the treatment of acute myelogenous leukaemia.     

Clinical studies on aclacinomycin A cardiotoxicity in adult patients with acute non lymphoblastic leukaemia

Nine patients with acute non lymphoblastic leukaemia (ANLL) were treated with Aclacinomycin in the doses of 20 mg/m2/day x 7 days in 30' lasting intravenous infusion and Cytosin arabinosid 100 mg/m2/day x 7 days in continuous infusion as well as control group consisted of 30 healthy people were examined by means of 24 hrs Holter ECG monitoring and ultrasonocardiography (UCG) to evaluate the influence of Aclacinomycin A (Aclaplastin - Behring) on cardiac rhythm and function. The UCG and Holter examinations were performed before Aclacinomycin and after 7-10 days from the beginning of the therapy. There were no statistical differences between the results of UCG examination in Aclacinomycin-treated group before the therapy and the control group. A slight nonsignificant decrease in left ventricular stroke volume and ejection fraction were observed after Aclacinomycin. Cardiac index decreased after the therapy (p less than 0.05) but was of normal value. The only true significant (p less than 0.001) decrease was observed in the contractility of cardiac fibres but the cardiac failure was not observed. No alterations in left ventricular posterior wall and intraventricular septum thickness were found. The effusion to pericardium was observed in 2 pts in the initial study and in 1 of them also after the therapy. The obtained results supported the clinical observations that Aclacinomycin A is promising agent for the treatment of ANLL because of its low cardiotoxicity.     

How should we talk about acute leukaemia to adult patients and their families?

Problems of communication with patients with acute leukaemia and their families were explored by interviewing the next of kin of 26 patients, six of whom were still alive. In all but two cases the diagnosis had been disclosed to the relatives before the patient, but almost one-third of the relatives were not entirely satisfied with the way in which the diagnosis was presented. Medical prognostications at these initial interviews were, on the whole, regarded as being realistic by the relatives. Nine patients learnt of their diagnosis at an early stage, but relatives were undecided whether patients should be told of the diagnosis in explicit terms. Patients often established complete dependence on the hospital and its staff and had difficulties in relating to their own general practitioners while at home during their illness. Social chats were preferred by the relatives rather than regular progress reports from the doctors.     

Clustering of childhood acute leukaemia

The EUROCLUS Project is a collaborative endeavour in which incidence data for 13 351 cases of childhood leukaemia (CL) diagnosed between 1980 and 1989 in 17 countries were referenced to 26 425 small geographic areas and tested for evidence of spatial clustering. A second objective of EUROCLUS was to determine whether clustering of CL was associated with community demographic features and/or proximity to putative environmental hazards. The results show statistically significant evidence of clustering, but the magnitude is small (extra-Poisson variability = 1.65% of Poisson variability). Patterns of clustering are associated with population density and other demographic features which could indicate variations in opportunity for exposure to common infections. There is no consistent evidence that clusters are associated with proximity to nuclear facilities or other putative environmental hazards. Received: 10 January 1998 / Accepted in revised form: 10 February 1998     

Apoptotic index, Fas and bcl-2 in initial and relapsed childhood acute lymphoblastic leukaemia

Seventy-two samples with initial and 23 samples with relapsed childhood acute lymphoblastic leukaemia (ALL) were investigated for apoptotic index (AI) and for Fas expression. AI was determined by DNA-fragmentation using deoxynucleotidyl terminal transferase and Fas expression by immunocytochemistry. bcl-2 mRNA expression was measured in 50 initial and 20 relapsed ALL. The patients were discriminated in groups with low and high AI, Fas-protein expression and bcl-2mRNA expression by the mean value. AI was higher in relapsed than in initial ALL. The mean survival was significantly higher in patients with low AI ( p= 0.034). This was also true for the relapse-free interval, however, this result was borderline significant ( p= 0.064). AI was directly correlated with expression of Fas and inversely correlated with bcl-2 mRNA expression. These results suggest that Fas and - with limitations - bcl-2 may influence the apoptotic process in childhood ALL and that enhanced apoptotic activity predicts poor prognosis.     

Expression and distribution of aphidicolin-induced fragile sites in chronic myeloid leukaemia, acute lymphocytic leukaemia and acute myeloid leukaemia

New information is revealed concerning the frequency of expression and distribution of aphidicolin-induced fragile sites in eight leukaemic patients, namely, four chronic myeloid leukaemic patients (CML), three acute lymphocytic leukaemic (ALL) patients, and one acute myeloid leukaemic (AML) patient. The cytogenetic data demonstrate a statistically significant (p less than 10(-6] increase in the frequency of aphidicolin-induced fragile sites in seven of the eight leukaemic patients compared with healthy age-matched and sex-matched controls. The chromosomal band locations of the aphidicolin-induced fragile sites from 400 metaphase spreads of these leukaemic patients reveal a nonrandom distribution in the karyotype. Some aphidicolin-induced fragile sites in these leukaemic patients were located at chromosome bands known to be induced specifically by folic acid, distamycin A, bromodeoxyuridine or azacytidine. The cross-induction of fragile sites in the leukaemic patients may be indicative of shared molecular homology in the sequence composition of nonrandom chromosomal DNA.