The influence of HLA-matched sibling donor availability on treatment outcome for patients with AML: an analysis of the AML 8A study of the EORTC Leukaemia Cooperative Group and GIMEMA

To determine whether patients with a HLA-identical sibling donor have a better outcome than patients without a donor, an analysis on the basis of intention-to-treat principles was performed within the framework of the EORTC-GIMEMA randomized phase III AML 8A trial. Patients in complete remission (CR) received one intensive consolidation course. Patients with a histocompatible sibling donor were then allocated allogeneic bone marrow transplantation (alloBMT), the patients without a donor were randomized between autologous BMT (ABMT) and a second intensive consolidation (IC2). 831 patients <46 years old and alive >8 weeks from diagnosis were included. HLA typing was performed in 672 patients. AlloBMT was performed during CR1 in 180 (61%) out of 295 patients with a donor. Another 38 patients were allografted: five in resistant disease, 14 during relapse and 19 in CR2. ABMT was performed in 130 (34%) out of 377 patients without a donor in CR1, in six (2%) patients during relapse and in 38 (10%) patients during CR2. The disease-free survival (DFS) from CR for patients with a donor was significantly longer than for patients without a donor (46% v 33% at 6 years; P = 0.01, RR 0.78, 95% confidence interval 0.63–0.96). The overall survival from diagnosis for patients with a donor was longer, but not statistically significant, than for patients without a donor (48% v 40% at 6 years; logrank P = 0.24). When patients were stratified according to prognostic risk groups, the same trend in favour of patients with a donor was seen for survival duration and the DFS remained significantly longer for this group of patients.     

Diurnal Variation of Proliferative Activity in the Human Bone Marrow

A study was made of the possible diurnal variation of proliferative activityof human bone marrow cells. A distinct diurnal variation of mitotic indiceswas present in 5 of 6 normal subjects. The average number of mitotic figuresper 1000 nucleated cells in the 6 subjects was 7.6 at 6:00 a.m., 9.5 at noon,10.5 at 6:00 p.m., and 13.1 at midnight. An assessment of diurnal variation inthe per cent of cells synthesizing deoxyribonucleic acid was made in 4 additional subjects by means of incubation of aspirated marrow samples withtritiated thymidine in vitro. By this method, no consistent variation of synthesiscould be found in erythroid precursor cells, but the per cent of myeloidprecursor cells incorporating the label was lowest at midnight in each subject.

Submitted on September 3, 1964 Accepted on November 5, 1964     

Leukokinetic studies. I. A method for labeling leukocytes with diisopropyl-fluorophosphate (DFP32)

1. A method has been presented by which granulocytes can be labeledin vivo with diisopropylfluorophosphate containing radioactive phosphorus.The leukocytes are isolated from blood by dextran sedimentation of erythrocytes and are then treated with gramicidin and lysolecithin to remove remaining red cells. Platelets are removed by differential centrifugation. Theisolated leukocytes are placed between two squares of scintillating plasticand counted with a scintillation counter.

2. Leukocytes essentially free of erythrocytes and platelets can be obtainedby the method outlined. The efficiency of the plastic scintillation countingmethod for radioactive phosphorus is about 74 per cent and leukocyte samples obtained from 20 ml. samples of normal blood can be counted with areproducibility of ±10 per cent.

3. The administration of 2 mg. of diisopropylfluorophosphate either intramuscularly or intravenously is without significant toxic side effects.

4. No evidence has been obtained that the label damages the leukocytes.

5. No evidence has been obtained that the label elutes from leukocytesunder the conditions of these studies.

6. Diisopropylfluorophosphate labels granulocytes for a brief period oftime following injection. The label is not reutilized after death of the cells.

Submitted on June 16, 1958 Accepted on July 17, 1958     

Granulocyte Kinetic Studies in Patients with Proliferative Disorders of the Bone Marrow

Granulocyte kinetic studies with DFP³² were done in four patients withchronic myelocytic leukemia, three patients with polycythemia vera, one patient with essential thrombocythemia, and one patient with persistent, unexplained granulocytosis. The increased blood granulocyte concentrationfound in the patients with polycythemia vera, essential thrombocythemia andunexplained granulocytosis was at least in part the result of increased granulocyte production. Precise calculations of granulocyte pool sizes and turnoverrates in the patients with chronic myelocytic leukemia were not possiblebecause of unresolved problems related to the non-uniform population ofmyeloid cells in the blood of these patients. However, within the limitationsof the method, a greater number of myeloid cells were turned over per daythrough blood than in normal subjects. The findings support the conceptthat a widespread disorder of marrow proliferation exists in chronic myelocytic leukemia, polycythemia vera, and essential thrombocythemia.

Submitted on December 20, 1962 Accepted on March 23, 1963     

Characteristics of Cell Proliferation in Four Patients with Untreated Acute Leukemia

The characteristics of proliferation of leukemic cells in four children withuntreated acute leukemia have been studied. In all four of these children apopulation of marrow leukemic cells was found which were dividing with ageneration time of about 15 to 20 hours. In two of these patients it waspossible to demonstrate that these dividing cells after one or more mitoticdivisions became smaller and stopped dividing. In all of these patients 70 percent or more of the leukemic cells of the marrow and almost all leukemic cellsof the blood were nonproliferative at the time of these studies. These nondividing cells would be relatively unaffected by chemotherapeutic agents designedto inhibit cell division.

Submitted on September 7, 1965 Accepted on January 31, 1965     

Congenital Pernicious Anemia with Coexistent Transitory Intestinal Malabsorption of Vitamin B12

Pernicious anemia with coexistent transitory intestinal malabsorption ofvitamin B₁₂ has been established in adults. In this report a child with congenitalpernicious anemia was documented to have had transitory selective intestinalmalabsorption of vitamin B₁₂. The diagnosis of congenital pernicious anemiawas established by the age of the patient, absence of intrinsic factor in gastricfluid, lack of antibodies to intrinsic factor and parietal cells, presence of HClin gastric fluid, and normal gastric biopsy. The xylose excretion test, 72-hourfecal fat determination, upper gastrointestinal series, and biopsy of the jejunumwere normal, but the Schilling test was abnormal with hog intrinsic factor ofknown potency on two occasions, indicating selective malabsorption of vitaminB₁₂. Seven months after therapy the Schilling test was still abnormal withoutintrinsic factor, but was normal with both human and hog intrinsic factor.The normal absorption with intrinsic factor after therapy is indicative that theselective malabsorption which was originally present was probably a consequence of the vitamin B₁₂ deficiency resulting from lack of intrinsic factor. Inpatients with abnormal radioactive vitamin B₁₂ absorption tests with administration of intrinsic factor, coexistent pernicious anemia must be excluded bydemonstration of intrinsic factor in gastric fluid.

Submitted on February 27, 1967 Accepted on April 13, 1967     

INVOLVEMENT OF HYPOGASTRIC AND PELVIC NERVES FOR CONVEYING CYSTITIS INDUCED NOCICEPTION IN CONSCIOUS RATS

PURPOSE: We determined the sites of the antinociceptive action of morphine in the experimental model of cyclophosphamide induced cystitis and investigated the afferent nerve fibers involved in nociception transmission originating from the bladder. MATERIALS AND METHODS: Cyclophosphamide (200 mg./kg.) given intraperitoneally was used to induce cystitis in male rats and their behavior was observed and scored. The effect of 2 mg./kg. systemic morphine given intravenously on cyclophosphamide induced behavioral modifications was tested when administered alone and after 100 microg. naloxone per rat given intrathecally at the L1 to L2 or L6 to S1 level. The spinal antinociceptive effect of morphine was also tested when administered intrathecally alone at 10, 100 and 200 microg. per rat at L1 to L2, alone at 100 microg. per rat at L1 to L2 or L6 to S1, alone at 100 microg. per rat at L1 to L2 and L6 to S1 simultaneously, alone at 200 microg. per rat at L1 to L2 and after 100 microg. naloxone per rat given intrathecally at L6 to S1 at 100 microg. per rat at L1 to L2. RESULTS: Cyclophosphamide induced marked modifications in the behavior of the rats, including a decreased breathing rate, eye closing and specific postures. Morphine given intravenously reversed these behavioral disorders and this reversal was completely prevented by pretreatment with intrathecal naloxone. A dose of 100 microg. per rat given intrathecally also reversed these behavioral disorders by about 25% at the L1 to L2 and L6 to S1 levels. In addition, a dose of 100 microg. morphine per rat administered intrathecally and simultaneously at L1 to L2 and L6 to S1 produced an effect equal to the sum of those observed when administered separately, that is about 50%, whereas morphine at an intrathecal dose of 200 microg. at L1 to L2 produced the same effect as 100 microg. given intrathecally at the same level or at L6 to S1 (25%). Also, 100 microg. naloxone per rat administered intrathecally at L6 to S1 prevented the effect of 100 microg. morphine at L1 to L2. CONCLUSIONS: These results confirm the previously reported antinociceptive effect of systemic morphine in this model of cyclophosphamide cystitis, suggest that this antinociceptive action is completely located at the spinal site and most importantly demonstrate by the pharmacological approach and behavioral analysis that nociceptive sensations originating from the bladder are conveyed by hypogastric and pelvic nerves in this cyclophosphamide cystitis model in the conscious rat.