Chronic myelogenous leukemia in the course of chronic lymphocytic leukemia: evidence for an independent clonal origin.

We report on a 69-year-old man who developed Ph-positive CML 6 years after the onset of B-cell CLL. When CML was diagnosed, both malignant cell populations were detected in bone marrow and peripheral blood. Peripheral leukocytes were fractionated by Ficoll-Hypaque density gradient, and cytogenetic and molecular studies were performed on mononuclear cell and granulocyte-enriched populations. Mononuclear cells were stimulated with either PHA or PWM. In PHA-treated cultures 76% of the metaphases were Ph-negative, while after PWM stimulation 87% were Ph-positive. A bcr rearrangement was observed in DNA from the granulocyte-enriched fraction, but not in mononuclear cells. On the contrary the IgH locus resulted in monoclonally rearranged DNA, only in peripheral blood mononuclear cells. These results indicate that the two neoplastic populations originated independently.

Giant neutrophils with increased peroxidase activity. Another evidence of dysgranulopoiesis in AIDS.

Using the automated hematologic analyzer Technicon H6000, which classifies leukocytes by their size and peroxidase activity, the authors have observed in nine patients with full-blown acquired immune deficiency syndrome (AIDS) a consistent increase in peroxidase content of circulating neutrophils. The increase in peroxidase activity was homogeneous in three patients (P less than 0.05). The most striking finding, however, was the occurrence of single abnormal neutrophils with peroxidase activity higher than the major neutrophil population (i.e., HPX [high peroxidase] cells). The importance of this phenomenon was correlated with the clinical status, higher HPX values being found in patients with more advanced disease. These instrumental observations were associated with the morphologic finding of atypical neutrophils, much larger than normal, with irregular nuclei and abundant cytoplasm filled with peroxidase-positive granulations. Such cells represent, in the authors' experience, the most common expression of dysgranulopoiesis in AIDS.

Long-term cytogenetic effects of antineoplastic treatment in relation to secondary leukemia.

Chromosome translocations are consistently present in leukemias and lymphomas and are likely to represent primary events in the development of these neoplasias. A study of conditions that predispose to leukemia could shed some light on the origin of these translocations and therefore help in clarifying their exact role in the process of neoplastic transformation. Based on this assumption, we studied a group of individuals treated with radiochemotherapy for previous lymphoma and who were at increased risk of developing a secondary leukemia. The group comprised 14 Hodgkin's disease patients, 11 non-Hodgkin's lymphoma patients, and 13 controls. The patients were in remission and had been off therapy for at least 6 months. Chromosomes were studied from phytohemagglutinin (PHA)-stimulated peripheral lymphocytes and from bone marrow cells by the direct method and after short-term cultures (72 hours). The latter were also exposed to 5-bromodeoxyuridine (BrdU). Metaphases were scored for chromosome breaks, gaps, and other rearrangements. The percentage of gaps and breaks was significantly higher in patients than in controls. The difference was induced by BrdU and was apparent in bone marrow cells, but not in peripheral lymphocytes. We conclude that individuals exposed to the action of mutagenic agents (radiochemotherapy) have an increased chromosome instability that could be related to their increased risk of developing a secondary leukemia.

Serum lactate dehydrogenase isoenzyme pattern in non-Hodgkin's lymphomas.

Serum lactate dehydrogenase (S-LDH) and its isoenzyme pattern were assayed in 63 non-Hodgkin's lymphoma (NHL) patients, 37 at diagnosis, 15 at relapse and 11 in complete remission (CR). S-LDH in NHL patients with active disease was higher than in normal subjects and CR patients (p less than 0.001). Among the isoenzymes, LDH-2 and LDH-5 showed no remarked differences; LDH-1 was reduced and LDH-3 and LDH-4 raised in comparison to the normal group (p less than 0.001). S-LDH levels and isoenzymes 1 and 4 were influenced by the stage, the histological subgroup and by the presence of general symptoms. In fact, cases in stage IV, with "high-grade malignancy" and with general symptoms, had higher S-LDH levels and more evident LDH-1 and LDH-4 changes than the other stages, the other histopathological subgroups and the cases classified as "A". S-LDH was the same as in normal subjects in the "low-grade" and "intermediate-grade" malignancies as was LDH-1 in stage II and LDH-4 in stages II and III, in "low-grade" malignancy and in the A cases. In contrast, LDH-3 was always high, with no significant difference in relation to the variables considered. Thus, in NHL, LDH-3 seems to be a reliable marker of the presence of the disease in any case, whereas S-LDH is more related to the spread of the lymphoma.

Automated measurement of red blood cell microcytosis and hypochromia in iron deficiency and beta-thalassemia trait.

Some routine red blood cell (RBC) measurements and indexes (count, mean volume, volume dispersion, and mean hemoglobin [HGB] concentration) can be used to differentiate iron deficiency from heterozygous beta-thalassemia. A number of formulas that incorporate two or more of these measurements have been described to amplify such differences. The H*1 hematology analyzer directly measures volume and HGB concentration of individual RBCs. We have assessed the diagnostic usefulness of conventional and new RBC measurements provided by the H*1 on a learning data set that comprised 119 patients with iron deficiency and 172 patients with beta-thalassemia trait, both untreated and uncomplicated. The most striking finding was the inverse behavior of percentages of microcytes (volume, less than 60 fL) and hypochromic RBCs (HGB concentration, less than 280 g/L) in the two conditions. In 162 of 172 patients with beta-thalassemia trait, the percentage of microcytes (mean, 33.1%; central 95th percentile range, 9.2% to 54.5%) was higher than the percentage of hypochromic RBCs (mean, 13.9%; central 95th percentile range, 1.7% to 24.7%). In 105 of 119 patients with iron deficiency, on the contrary, the percentage of hypochromic cells (mean, 34.6%; central 95th percentile range, 9.7% to 73.1%) was higher than the percentage of microcytes (mean, 12.8%; central 95th percentile range, 1.7% to 29.6%). The ratio between the percentage of microcytes and the percentage of hypochromic cells provided by the H*1 (microcytic-hypochromic ratio) was useful in differentiating the two types of microcytic anemia: with the use of a discriminant value of 0.9, the discriminant efficiency of the microcytic-hypochromic ratio was 92.4% (95% confidence interval, 88.8% to 95.2%), higher than that of the five previously described discriminant formulas and simple RBC measurements. When assessed on a test data set that comprised 149 unselected cases of microcytic anemia, a microcytic-hypochromic ratio lower than 0.9 demonstrated high sensitivity (94.0%), specificity (92.3%), and predictive value (94.0%) for the presence of iron-deficient erythropoiesis in patients with isolated iron deficiency, polycythemia vera treated by phlebotomy, and iron deficiency complicating heterozygous thalassemia. In conclusion, our results showed that iron-deficient erythropoiesis is characterized by the production of RBCs with a severely decreased HGB concentration, while microcytes of beta-thalassemia trait are generally smaller, with a more preserved HGB concentration. Such properties, as assessed by the H*1 hematology analyzer, are very useful in distinguishing these two common types of microcytic anemia.

Sequential therapy with lithium in chemotherapy-induced vomiting.

Having noticed psychotic traits in some patients showing incoercible vomiting due to antineoplastic drugs we have thought of establishing a therapy with lithium in the days preceding the therapeutic cycle in order to reduce the emetic events. The effectiveness of lithium carbonate (600 mg/mq p.o./day for one week) in the prevention or reduction of vomiting induced by antiblastic therapy has been checked in comparison with metoclopramide and domperidone in 40 patients. In the group pretreated with lithium, 80% of the cases showed favorable results. In the control groups, on the contrary, the efficacy of the antiemetic therapy has shown to be of lesser importance (55%). The undesirable side-effects of lithium appear to be irrelevant. We therefore think that pretreatment with lithium may become, in selected cases not affected by traditional antiemetics, of great importance in the control of emetic symptomatology.

Chronic myeloid leukemia with monoclonal gammopathy terminating in myeloid crisis and immunoblastic lymphoma.

A patient, with chronic myeloid leukemia and IgA monoclonal gammopathy, who contemporaneously developed myeloid blast crisis and immunoblastic lymphoma is reported. Cytogenetic studies showed complex chromosome abnormalities concerning chromosomes 8, 14 and 22, other than the Ph chromosome. A possible relationship between the emergence of immunoblasts from slow proliferating lymphoplasmacytoid cells, myeloid blasts crisis and chromosomal changes is discussed.

Depletion lymphocytapheresis in chronic lymphocytic leukemias: criteria for predicting which patients will respond to treatment.

Depletion lymphocytapheresis therapy using the continuous-flow separator was used in twenty-two cases of chronic lymphocytic leukemia. Applying the Montserrat scoring system, a prediction can be made, to a good approximation, whether the patient will be a "responder" or a "non-responder" to leukapheresis. Criteria have been formulated for decisions on when depletion lymphocytapheresis is absolutely or only relatively indicated, on the basis of total scores from 2 through 6 using the Montserrat criterion.

[Gaucher's disease: hemorrhagic diathesis and term pregnancy with the birth of a malformed fetus]. / Morbo di Gaucher: diatesi emorragica e gravidanza conclusa con la nascita di un feto malformato.

A patient suffering from Gaucher's disease observed for the first time five years age while in pregnancy and observed again during a second pregnancy ended with the delivery of a malformed female infant is presented. Laboratory findings of this case and their correlation with the clinical manifestations of this disease are underlined. Occurrence of foetal malformations in this disease is also discussed.

Elevation of susceptibility to ozone-induced acute tracheobronchial injury in transgenic mice deficient in Clara cell secretory protein.

Increases in Clara cell abundance or cellular expression of Clara cell secretory protein (CCSP) may cause increased tolerance of the lung to acute oxidant injury by repeated exposure to ozone (O3). This study defines how disruption of the gene for CCSP synthesis affects the susceptibility of tracheobronchial epithelium to acute oxidant injury. Mice homozygous for a null allele of the CCSP gene (CCSP-/-) and wild type (CCSP+/+) littermates were exposed to ozone (0.2 ppm, 8 h; 1 ppm, 8 h) or filtered air. Injury was evaluated by light and scanning electron microscopy, and the abundance of necrotic, ciliated, and nonciliated cells was estimated by morphometry. Proximal and midlevel intrapulmonary airways and terminal bronchioles were evaluated. There was no difference in airway epithelial composition between CCSP+/+ and CCSP-/- mice exposed to filtered air, and exposure to 0.2 ppm ozone caused little injury to the epithelium of both CCSP+/+ and CCSP-/- mice. After exposure to 1.0 ppm ozone, CCSP-/- mice suffered from a greater degree of epithelial injury throughout the airways compared to CCSP+/+ mice. CCSP-/- mice had both ciliated and nonciliated cell injury. Furthermore, lack of CCSP was associated with a shift in airway injury to include proximal airway generations. Therefore, we conclude that CCSP modulates the susceptibility of the epithelium to oxidant-induced injury. Whether this is due to the presence of CCSP on the acellular lining layer surface and/or its intracellular distribution in the secretory cell population needs to be defined.

[Value of CT and lymphography in the staging of lymphomas (author's transl)]. / Ruolo della tomografia computerizzata e della linfografia nello staging dei linfomi.

The contribution of CT is evaluated in association with conventional diagnostic investigations in the staging of lymphomas. The data obtained by means of CT are an integration of those obtained by means of lymphography. On the other hand CT offers elements which cannot be obtained by others methods, especially concerning the retroperitoneal region. This diminishes the average of patients who must undergo laparoscopy and laparotomy. Moreover CT allows to follow the evolution of the disease and to control the results of the treatment.

Disseminated intravascular coagulation in acute promyelocytic leukemia. Possibility of treatment with glucocorticoids at high doses.

Seven patients affected by acute promyelocytic leukemia and presenting a hemorrhagic syndrome with hypofibrinogenemia have been treated with Daunorubicin and high doses of Glucocorticoids. In three out of the seven patients we also added an antiprotease agent (Trasylol, Bayer). Hemorrhagic diathesis has been controlled in all patients but one without massive platelet transfusions. A complete hematologic remission with prolonged survival was achieved in five patients.

Automated cytochemistry in acute leukemias. A new approach to the FAB classification based on cell distribution pattern.

Automated cytochemistry by the Hemalog D carries out leukocyte differential counts in a continuous flow mode on whole blood EDTA-collected samples, through the optical measurement of enzyme activity and cell size. This operative principle thus parallels the basis of the FAB classification of acute leukemias. Although the cell classification logic system can be misled by the heterogeneity of most leukemic populations, valuable qualitative information may be obtained from the x-y oscilloscope display. This provides a true morphological representation of the leukemic cell distribution. According to such a preliminary assumption, each FAB subtype of acute leukemias is shown to reproduce a peculiar image of cellular distribution when analyzed by the Hemalog D.

Altered pulmonary response to hyperoxia in Clara cell secretory protein deficient mice.

Clara cell secretory protein (CCSP) is an abundant component of the extracellular lining fluid of airways. Even though the in vivo function of CCSP is unknown, in vitro studies support a potential role of CCSP in the control of inflammatory responses. CCSP-deficient mice (CCSP -/-) were generated to investigate the in vivo function of this protein (13). In this study, we used hyperoxia exposure as a model to investigate phenotypic consequences of CCSP deficiency following acute lung injury. The pathologic response of the mouse lung to hyperoxia, and recovery of the lung, include inflammatory cell infiltrate and edema. Continuous exposure to > 95% O2 was associated with significantly reduced survival time among CCSP -/- mice as compared with strain-, age-, and sex-matched wild-type control mice. Differences in survival were associated with early onset of lung edema in CCSP -/- mice as compared with wild-type controls. To further investigate these differences in response, mice were exposed to > 95% O2 for either 48 h or 68 h with one group receiving 68 h of hyperoxia followed by room-air recovery. Lung RNA was characterized for changes in the abundance of cytokine messenger RNA (mRNA) using a ribonuclease (RNase) protection assay. After 68 h of hyperoxia, interleukin-6 (IL-6), IL-1beta, and IL-3 mRNAs were 14-, 3-, and 2.5-fold higher, respectively, in CCSP -/- mice than in similarly exposed wild-type control mice. Increased expression of IL-1beta mRNA in hyperoxia-exposed CCSP -/- mice was localized principally within the lung parenchyma, suggesting that the effects of CCSP deficiency were not confined to the airway epithelium. We conclude that CCSP deficiency results in increased sensitivity to hyperoxia-induced lung injury as measured by increased mortality, early onset of lung edema, and induction of proinflammatory cytokine mRNAs.

Megathrombocytes, platelet regeneration time and platelet associated IgG in idiopathic thrombocytopenic purpura and in thrombocytopenia associated with chronic liver disease.

Percentage of megathrombocytes, platelet regeneration time (PRT) and platelet-Associated IgG (Pl-A-IgG) were investigated in 12 patients with clinical features consistent with idiopathic thrombocytopenic purpura and in 11 patients with thrombocytopenia associated with chronic liver disease. Bone marrow smears were also examined and megakaryocytes classified into stages I-III according to the current principle. Of 12 patients with idiopathic thrombocytopenic purpura the percentage of megathrombocytes was increased in 9, PRT reduced in 10, and Pl-A-IgG increased in 8 patients. A statistically significant correlation was found between the percentage of megathrombocytes and the level of Pl-A-IgG. A slight correlation was also found between PRT and the percentage of megathrombocytes, while a significant correlation was found between megakaryocytes in stage I and the percentage of megathrombocytes, suggesting that growth of megakaryocytes probably contributes to platelet heterogeneity. In patients with thrombocytopenia and chronic liver disease, the percentage of megathrombocytes was in the normal range. A moderately increased level of Pl-A-IgG was found only in patients with active chronic hepatitis, PRT was reduced only in a few patients, while most of them also showed an increase of Pl-A-IgG.

Electromyography for the investigation and early diagnosis of scoliosis.

A clinically normal 5 year old child with a family history of scoliosis was studied. Electromyography of the thoracic and lumbar erector spinae muscles and roentgenography of the spine were both done on two separate occasions, six months apart. On the first electromyographic investigation, moderate predominance of activity was found over the left thoracic erector spinae muscles. At that time, spinal roentgenography showed normal results, whereas six months later left thoracolumbar scoliosis was evident. It was concluded that electromyographic investigation is useful for the early diagnosis of scoliosis in school-age children.

Identification of blast cells in peripheral blood through automatic assessment of nuclear density: a new tool for monitoring patients with acute leukaemia.

The Technicon H*1 haematology system is provided with a new method for basophil count, neutrophil lobularity assessment and detection of blasts in peripheral blood through automated measurement of nuclear density. We compared the results of the H*1 blast flag with those of the microscope examination in 131 peripheral blood samples from 43 patients with acute leukaemia in different phases of their disease, to determine the degree of sensitivity and specificity of the system in this setting. In six patients at diagnosis or in overt relapse, all having large percentages of blasts at the manual differential count, a typical deformation of the profile of the mononuclear cell population on the display was consistently observed, regardless of the morphological subtype of leukaemic cell which was involved. Amongst 34 samples with 4-95% morphologically recognizable blasts on the peripheral blood film, the sensitivity of the system was 100% with no false negatives at all. In 43 samples, on the other hand, the H*1 blast flag was positive in the absence of any morphological evidence of blasts on the smear. These 'false positives', however, were always obtained from leucopenic patients who had more than 12% blast infiltration in the bone marrow, compared to the 0-6% value that was found in patients with a negative H*1 blast flag. These results suggest that the H*1 system is a highly sensitive tool for the detection in peripheral blood of even small concentration of leukaemic cells, which escape morphological identification.