bluuebunny/crossref_metadata_2025 huggingface dataset:
Thrombocytopathy in Preleukaemia: Association with a Defect of Thromboxane A2 Activity
Authors: Russell N. H. , Keenan J. P. , Bellingham A. J.
Published: March 2008

Summary. Platelet aggregation and the platelet prostaglandin pathway have been investigated in two patients with preleukaemic states who had a haemorrhagic tendency but a normal platelet count. In both patients platelet aggregation induced by collagen, adenosine diphosphate (ADP) and arachidonic acid (AA) were abnormal. Malonyldialdehyde (MDA) production from exogenous AA was normal in both patients thus excluding cyclo‐oxygenase deficiency. The platelet aggregating and rabbit aorta contracting activities of thromboxane A2 (TxA2) were very low in both patients. Production of thromboxane B2 (TxB2) assessed by thin layer chromatographic separation of the metabolites of [I‐14C]AA and by radioimmunoassay, was normal. These abnormalities of platelet function appear to be due to the production of TxA2 with a low biological activity.
Response to Treatment in Acute Non‐lymphatic Leukaemia: Prognostic Value of Colony Forming and Colony Stimulating Capacities of Bone Marrow and Blood Cells Compared to Other Parameters
Authors: Beran Miloslav , Reizenstein Peter , Uden Ann‐Mari
Published: March 2008

Summary.

Growth of bone marrow and mononuclear white blood cells (MWBC) in soft‐agar cultures was studied in 26 patients with untreated acute non‐lymphocytic leukaemia (ANLL). Marrow and MWBC from 30 healthy volunteers served as controls. All ANLL patients revealed an abnormal growth in vitro. Patients with an increased number of clones in marrow cultures and large cluster predominance (‘excessive growth’) responded poorly to therapy with only one of 10 patients entering remission. On the contrary, only two of the 15 patients with a decreased clone number (‘low growth’) failed to achieve remission. The number of colonies and clusters in both bone marrow and blood cultures was significantly lower at presentation in patients who later entered remission than in those who did not. The correlations between the number of colonies and clusters in the blood and the marrow cultures were statistically significant. No significant correlations were found between prognosis or colony formation, on one hand, and the production of colony stimulating activity (CSA), by bone marrow and blood cells of ANLL patients, on the other. Nor could such correlations be found between prognosis, blood cell counts, and age. It is concluded that the growth pattern of both bone marrow and circulating colony forming cells is of value in predicting the response to cytostatic drugs and particularly in selecting patients with a high probability to respond poorly to current cytostatic regimes.
Polymorphonuclear (PMN) Function after Small Intestinal Shunt Operation for Morbid Obesity
Authors: Palmblad Jan , Hallberg Dag , Engstedt Lars
Published: March 2008

Summary.

The possible influence on blood polymorphonuclear (PMN) granulocyte functions of the small intestinal shunt operation for obesity was studied in 10 massively overweight patients. They were investigated prior to operation and for 9 months afterwards, when they had lost on average 32 kg body weight. Preoperatively they showed reduced PMN bactericidal capacity and increased PMN adherence compared with controls of normal weight. During the first 2–4 months postoperatively all patients displayed a gradually increasing bactericidal capacity, which then reached levels similar to the controls and remained so for the rest of the follow‐up period. This enhancement was more easily assessed by a new in vitro assay in which each PMN was provided with 30–40 bacteria, than by a standard assay using 2–4 bacteria per granulocyte. PMN adherence decreased during the first postoperative months and then returned to preoperative levels. The changes in PMN functions were not statistically related either to each other or to the continuous loss of body weight. Thus, impairment of PMN killing function occurring in extremely obese patients became normalized after small bowel shunt operation, while the high adherence remained unchanged.
Origin of Human Bone Marrow Fibroblasts
Authors: Golde D. W. , Hocking W. G. , Quan S. G. , Sparkes R. S. , Gale R. P.
Published: March 2008

Summary.

We investigated the origin of bone marrow fibroblasts in three bone marrow transplant recipients with aplastic anaemia and leukaemia who received grafts from HLA‐identical siblings of opposite sex. The patients were conditioned for transplantation with high doses of cytotoxic drugs and 300–1000 rads total body irradiation. After transplantation, bone marrow cells wrere cultured in T flasks for 3 weeks and the adherent cells were then trypsinized and passaged weekly. After several passages the cells had the typical morphology and growth pattern of fibroblasts. Metaphases from these cells were all of recipient sex type. In contrast, haematopoietic cells and lymphocytes obtained at the same time were of donor sex type. Our findings indicate that the human bone marrow fibroblast is not derived from a precursor common to haematopoietic cells or lymphocytes. The bone marrow fibroblast appears to be a mesenchymal cell, unrelated to haematopoietic stem cells, which is capable of in vitro proliferation after as much as 1000 rads of total body irradiation.
Polycythaemia Vera: in Vitro Studies of Circulating Erythroid Progenitors
Authors: Lacombe C. , Casadevall N. , Varet B.
Published: March 2008

Summary.

Peripheral blood mononuclear cells from healthy subjects, patients with secondary polycythaemia and patients with polycythaemia vera (PV) were cultured using the plasma clot technique, and large erythroid colonies or bursts were scored after 14 d. Erythroid bursts appeared without addition of erythropoietin in all 20 patients with PV but never in normal subjects (n=10) nor in patients with secondary polycythaemia (n=6). The erythropoietin dose‐response curves in six PV patients were characterized by an initial plateau followed by a nearly normal response. This suggests the coexistence of two populations of erythroid progenitors in PV, one abnormally sensitive, the other normally responsive to erythropoietin. In nine patients the ratio of spontaneous bursts to bursts induced by high doses of erythropoietin from the blood on day 14 was identical with the ratio of spontaneous colonies to colonies induced by high doses of erythropoietin from the bone marrow of the same patients on day 7. This argues against the hypothesis that spontaneous erythroid bursts are derived from erythroid progenitors (BFUE) abnormally sensitive to erythropoietin. Moreover blood mononuclear cells from healthy subjects were found to give rise to erythroid bursts when erythropoietin addition was delayed until 6 d in culture. This is consistent with the concept that erythropoietin is not normally required in vivo for the differentiation of early (BFUE) to late (CFUE) erythroid progenitors. These findings suggest that in PV all BFUE are normally responsive to the physiological factor(s) different from erythropoietin and responsible for their differentiation into CFUE, and that the abnormal response to erythropoietin, distinctive of a population of erythroid progenitors, is expressed only at the most mature (CFUE) level.
Terminal Transferase Enzyme Assay and Immunological Membrane Markers in the Diagnosis of Leukaemia: a Multiparameter Analysis of 300 Cases
Authors: Janossy G. , Hoffbrand A. V. , Greaves M. F. , Ganeshaguru K. , Pain Carol , Bradstock K. F. , Prentice H. G. , Kay H. E. M. , Lister T. A.
Published: March 2008

Summary.

Multiparameter analyses have been carried out with recently developed enzyme and membrane markers in 300 patients with various leukaemias including ALL, AML, but excluding Ph1 positive leukaemias.

TdT enzyme levels were particularly valuable in the differential diagnosis of adult acute lymphoid and myeloid leukaemias. The levels were raised in 108 (94%) of the 115 patients who were considered to be non‐T, non‐B ALL on membrane marker and morphological analysis; all seven cases giving negative TdT results in this group were young children. Unexpectedly high levels were seen only in three (4.1%) of 73 cases of acute myeloid leukaemia verified by histochemistry and membrane markers. Anti‐ALL serum was a most useful reagent in childhood leukaemias but blasts from 19 patients (10% of childhood ALL cases and 29% of adult ALL cases) failed to react with the serum in spite of TdT positivity. Strongly ALL+ blasts were seen only in non‐T, non‐B ALL and some undifferentiated leukaemias. Weakly ALL+ blasts were seen in seven of 32 cases of thymic ALL (Thy‐ALL) but in other respects these blasts expressed Thy‐ALL features, such as strong reactivity with anti‐T cell (HuTLA) serum, negativity with anti‐Ia‐like serum and raised TdT. The combination of tests was particularly useful in 32 cases of undifferentiated leukaemia: in 10 of these cases TdT positivity indicated the probable ‘lymphoblast’, nature of blast cells: the remaining 22 cases remained unclassifiable with the markers used. The analysis revealed other interesting variant forms of leukaemias.
Cytobiological and Clinical Aspects of Tissue Mast Cell Leukaemia
Authors: Coser P. , Quaglino D. , Pasquale A. De , Colombeiti V. , Prinoth O.
Published: March 2008

Summary. The authors describe the results of a series of cytochemical, autoradiographic, cytophotometric and immunological investigations carried out in a case of tissue mast cell leukaemia. Leukaemic mast cells showed certain distinctive cytochemical features, amongst which an intense periodic acid‐Schiff (PAS) reaction, sensitive to amylase digestion, strong naphthol AS‐D chloroacetate esterase (NASDCE), intense lactate dehydrogenase (LD) activity. Proliferative activity, determined autoradiographically with 3H‐dT, was considerably low and was mainly confined to the larger cells. Also uridine and leucine incorporation were markedly reduced. Microdensitometry disclosed that the mast cell population was mainly arrested in the G1 phase. Because of previous attempts to destroy selectively neoplastic tissue mast cells with sheep antihuman IgE serum, a search for surface bound IgE was carried out, but gave a negative result. Possible therapeutic approaches are considered in the light of previous clinical experience and on the basis of the results of the kinetic and metabolic studies.
Determination of Alpha Thalassaemia Phenotypes by Messenger RNA Analysis
Authors: Hunt D. M. , Higgs D. R. , Old J. M. , Clegg J. B. , Weatherall D.J. , Marsh G. W.
Published: March 2008

Summary. The possibility of using α/β globin messenger RNA (mRNA) ratios to distinguish between the carrier states for different forms of a thalassaemia has been explored, α/β globin mRNA ratios were determined in the red cells of a series of normal individuals and in members of four Cypriot families, one Thai and one Chinese family in which at least one person has haemoglobin H disease. It was found that there was a clear distinction in the ratios between normals, α thalassaemia 1 carriers, α thalassaemia 2 carriers, and those with haemoglobin H disease. This method should be a valuable addition to haematological analysis, haemoglobin synthesis and restriction mapping of DNA for the further elucidation of the genetics of α thalassaemia.
Lymphocyte Density Distribution Profile and Spontaneous Transformation Related to the Stage of Hodgkin's Disease
Authors: De Pauw B. E. , Th. Wagener D. J. , Smeulders J. B. J. M. , Geestman E. J. M. , Wessels J. M. C. , Haanen C.
Published: March 2008

Summary

. Density distribution profiles of peripheral lymphocytes were made in 41 untreated patients with Hodgkin's disease and in 14 normal controls. The fraction of lymphocytes with a low specific gravity in patients was significantly increased in comparison to normal controls (P= 0.0007). These low density lymphocytes in Hodgkin's disease patients showed an elevated spontaneous 3H‐thymidine incorporation compared to normal controls (P < 0.0001). In 78% of the patients the spontaneous DNA synthesis of the low density lymphocytes was above the normal range. A positive correlation (P= 0.002) was found between the spontaneous 3H‐thymidine uptake and the stage of the disease.
Haematological Aspects of Antenatal Diagnosis for Thalassaemia in Britain
Authors: Matsakis M. , Berdoukas V. A. , Angastiniotis M. , Mouzouras M. , Ioannou P. , Ferrari M. , Modell B. , Fairweather D. V. I. , Ward R. H. T. , Loukopoulos D. , Sakarellou N.
Published: March 2008

Summary.

The results are described of 200 antenatal diagnostic tests for haemoglobinopathies performed on samples of fetal blood obtained during the second trimester of pregnancy. Haemoglobin A synthesis in the fetus was measured by incorporation of tritiated leucine in vitro and separation of the globin chains on CM23 columns. The range of HbA synthesis detected was 3.5–8.0% in normal fetuses, 2.0–5.0% in fetuses with thalassaemia trait, and less than 1.6% in fetuses with thalassaemia major. There were eight cases in which other haemoglobinopathies were diagnosed. 29% of the pregnancies were terminated because thalassaemia major was diagnosed, and 9.5% of the remaining healthy fetuses were lost for obstetric reasons. Follow up has been possible for 96% of the 124 surviving babies and three misdiagnoses have come to light; one false positive (0.5%) and two false negatives (1%). These figures represent a first effort at antenatal diagnosis for haemoglobinopathies and it is likely that they will improve with the passage of time.
Five Families with Homozygous δ‐Thalassaemia in Japan
Authors: Yasukawa Masaki , Saito Shuji , Fujita Shigeru , Ohta Yoshiro , Ikeda Kyoichi , Matsumoto Isao , Kobayashi Yuzuru
Published: March 2008

Summary.

Five families with δ‐thalassaemia discovered in Ehime, Japan, are presented. The δ‐thalassaemia was associated with a slight elevation of the level of Hb F in two families and with normal Hb F levels in three. Complete absence of HbA2 was found in the homozygous probands. No abnormal clinical or haematological findings were noted in the individuals with δ‐thalassaemia.
Clinical and Immunologic Spectrum of Plasmacytoid Lymphocytic Lymphoma without Serum Monoclonal IgM
Authors: Levine Alexandra M. , Lichtenstein Alan , Gresik Mary V. , Taylor Clive R. , Feinstein Donald I. , Lukes Robert J.
Published: March 2008

Summary.

As first suggested by Lukes & Collins (1979, and recently confirmed in the literature, the pathological lesion associated with Waldenström's macroglobulinaemia can be found in patients with serum IgG or IgA monoclonal proteins. To expand upon the clinical and immunologic spectrum of this lesion, we reviewed the records of 12 patients with identical tumour morphology, though lacking an IgM paraprotein. The median age of the patients was 62 years. In five patients the tumour secreted monoclonal IgG, while one secreted monoclonal IgA. One patient had normal serum protein electrophoresis, although monoclonal light chains were demonstrated on urinary protein electrophoresis. The remaining patients had no detectable monoclonal protein. Marker studies demonstrated the U cell origin of tumour; these findings were consistent with the monoclonal protein secreted. Clinically, these cases varied, from peripheral blood involvenient resembling chronic lymphocytic leukaemia, to typical lymphomatous processes, to presentations resembling multiple myeloma, reflecting the similarity of all of these conditions as varying expressions of neoplasia of the B cell system.
Decreased Association of 45Calcium with Platelets Unable to Aggregate due to Thrombasthenia or Prolonged Calcium Deprivation
Authors: Peerschke Ellinor I. , Grant Robert A. , Zucker Marjorie B.
Published: March 2008

Summary.

45Calcium uptake was studied with aspirin‐treated platelets that were gel‐filtered through a column of Sepharose 2B equilibrated with divalent cation‐free modified Tyrode's solution to remove readily exchangeable surface‐associated calcium. These platelets aggregated almost immediately when exposed to ADP, fibrinogen and at least 30 μm CaCl2. At this calcium ion concentration, 108 platelets took up 36.6 ± SEM 2.7 pmol of 45calcium within 1–2 min. The presence of ADP and fibrinogen did not affect the amount of calcium bound. Over 90% of this platelet‐associated calcium was removed by EDTA in 5 min suggesting that it was surface‐bound. Calcium uptake increased rapidly for 10 min, then more slowly for up to 2 h. At 60 min, maximal uptake was approached at CaCl2 concentrations between 250 and 300 μm when an average of 276 ± SEM 18 pmol of calcium was associated with 108 platelets. Only 50–60% of this calcium could be removed by EDTA in 5 min, and about 70% in 20 min, suggesting that some of it had been internalized. Platelets from two patients with thrombasthenia that were unable to aggregate took up 50% less calcium than platelets from normal volunteers. Similarly, platelets that had been incubated with EDTA at 37°C, pH 7.8 for 8 min lost the ability to aggregate despite recalcification and took up 40–60% less calcium than CaEDTA‐treated controls. Platelets from a patient with the Bernard‐Soulier syndrome aggregated and bound calcium normally. Thus the platelets’ ability to take up calcium after removal of surface‐associated calcium correlates with their ability to aggregate. Since thrombasthenic platelets and platelets rendered incapable of aggregating after prolonged calcium deprivation with EDTA do not bind fibrinogen, we postulate that some of the surface‐associated calcium normally binds to the fibrinogen receptors.
Immunochemical Analysis of Active and Inactive Antithrombin III
Authors: McKay E.J.
Published: March 2008

Summary.

Antithrombin III (AT) levels from normal and AT deficiency persons were measured by electroimmunoassay (EIA) and the rewlts compared with a chromogenic assay (S2238). Discrepant results were obtained when plasma and serum were compared using one antiserum, and therefore did not always relate to functional activity. Another antiserum, however, when used was capable of differentiating active AT from inactive AT complexed with its proteases and demonstrated close correlation with all samples tested (r=0.97). The specificity of the antisera and consequent anomalous results were elucidated when purified human thrombin was added to plasma samples and subsequently reanalysed. Quantitative differences observed when serum samples were compared by single radial immuno‐diffusion and electroimmunoassay with one antiserum, illustrates the fundamental principle differences between the two methods. These results give some insight as to why previous anomalies with AT immunoassays have occurred. They also indicate that plasma and not serum should be used as clinical test material. AT antisera should be capable of recognizing and distinguishing free AT from AT/protease complexes if the results obtained by electroimmunoassay are to correlate with functional activity.
Ultrastructural Studies of Erythropoiesis in β‐Thalassaemia Trait
Authors: Wickramasinghe S. N. , Hughes Madeleine
Published: March 2008

Summary.

The erythropoietic cells of six cases of β‐thalassaemia trait were studied by electron microscopy and electron microscope autoradiography. Intracytoplas‐mic and intranuclear α‐chain precipitates were found in some late polycliromatic erythroblasts and intracytoplasmic precipitates were found in several marrow reti‐culocytes. This provides direct morphological evidence of unbalanced globin chain synthesis in the marrow. Several of the polychromatic erythroblasts and marrow reticulocytes contained autophagic vacuoles and showed a variety of other dysery thropoietic changes. Erythroblast profiles containing moderate quantities of precipitated α‐chains usually suffered from a marked depression of protein synthesis. The proportions of marrow cells containing α‐chain precipitates and displaying dyserythropoietic changes varied considerably from patient to patient. It is proposed that this variation largely reflects variations in the proteolytic capacity of the erythropoietic cells in different individuals and leads to different degrees of ineffective erythropoiesis in β‐thalassaemia trait.
Platelet‐Activating Factor (PAF‐Acether) Secretion from Platelets: Effect of Aggregating Agents
Authors: Chignard M. , Couedic J. P. Le , Vargaftig B. B. , Benvenisie J.
Published: March 2008

Summary.

Platelet‐activating factor is a 1.0‐alkyl‐2 acetyl analogue of phosphatidylcholine (PAF‐acether) which triggers platelet aggregation independently from ADP release and thromboxane A2 (T×A2) formation. PAF‐acether was described initially as being secreted by rabbit basophils during an immunological challenge. We have now found that it is also formed by washed rabbit platelets stimulated by the calcium ionophore A23187, thrombin and collagen. These three agents are known to trigger platelet aggregation independently from the release of ADP and from the formation of T×A2. By contrast, ADP and arachidonic acid, the precursor of T×A2, which do not share these properties, and PAF‐acether itself, were unable to induce PAF‐acether formation. Our results suggest that PAF‐acether may be the mediator responsible for AIIP and T×A2 independent‐aggregation.
Impaired Thromboxane Production by Newly Formed Platelets after Aspirin Administration to Thrombocytopenic Rats
Authors: Dejana Elisabetta , Barbieri Brunelia , Cerletti Chiara , Livio Manuela , Gaetano Giovanni de
Published: March 2008

Summary.

The in vivo inhibitory effect of aspirin on platelet cyclo‐oxygenase is irreversible and lasts for the entire platelet life‐span. Reappearance of cyclo‐oxygenase activity in blood after aspirin has been proposed as a measure of the formation of new platelets and as an indirect indicator of platelet survival. A delay of 24–72 h in recovery, however, has been observed and it has been suggested that aspirin might also inhibit megakaryocyte cyclo‐oxygenase. To test this possibility, aspirin (100 mg/kg) or saline were administered i.p. to rats made thrombocytopenic 2 h later (platelet count less than 5% of basal value) by a specific antiplatelet antiserum. Malondialdehyde (MDA) and thromboxane B2 (T×B2) production by platelets was measured by spectrophotometry and radioimmunoassay respectively, during the period of platelet count restoration. By 24 h after thrombocytopenia was induced, platelet count was about 15% of basal values in control and aspirin‐treated rats. However, while in controls MDA and T×B2 production was restored to about 20% of basal values, in aspirin‐treated rats less than 5% return of activity was detected. A marked difference between the two groups was still found 96 h after induction of thrombocytopenia, when platelet count restoration was similar. Since aspirin disappeared very rapidly from the circulation, the delay in recovery of cyclo‐oxygenase activity supports the hypothesis of a megakaryocyte effect of this drug.
Non‐spherocytic Haemolytic Anaemia in Mother and Son Associated with Hexokinase Deficiency
Authors: Newman P. , Muir A. , Parker A. C.
Published: March 2008

Summary.

The case history, laboratory findings and properties of the enzyme in a patient with hexokinase deficiency are reported. The mother had a haemolytic disorder of similar severity. In these cases the expression of the disease in both mother and son suggests a dominant mode of inheritance. This family further illustrates the necessity of taking the reticulocyte level into account when interpreting red cell enzyme levels.
Maturation Index: a Contribution to Quantification in the FAB Cla.ssification of Acute Leukaemia
Authors: Rhenen D. J. van , Verhulst J. C. , Huijgens P. C. , Langenhuijsen M. M. A. C.
Published: March 2008

Summary.

The French—American—British co‐operative study group (FAB) criteria were used to classify acute leukaemia in 194 adult patients. The dividing lines between FAB subgroups M1‐M2 and M5A—M5B appeared to be less well defined. In an effort to improve subtyping we introduce the maturation index (MI).

MI=promyelocytes/promonocytes/blast cells + promyelocytes/promonocytes/×100%.

Clearly distinct subgroups emerged after arrangement of cases according to MI values. Immature and rather mature cases in both myeloblastic and monoblastic leukaemia were divided at a MI of 4%. Assignment to M3, which is based essentially on morphological grounds, could be made on numerical grounds too, MI being at least 85% in all of our cases. A few cases showing M2 morphology and a positive α‐naphthylacetate esterase (ANE) staining should probably be categorized as M4. Application of the original FAB criteria resulted in disagreement of 14% between three investigators, while with our additional criteria the discrepancy was only 2%.
Heteromorphisms of the Philadelphia (Ph1) Chromosome in Patients with Chronic Myelogenous Leukaemia (CML). I. CLASSIFICATION AND CLINICAL SIGNIFICANCE
Authors: Verma Ram S. , Dosik Harvey
Published: March 2008

Summary In patients with chronic myelogenous leukaemia (CML), we have found the break points on the long arm of chromosome 22 (22q) are variable (heteromorphic or polymorphic). Consequently, the Philadelphia (Ph1) chromosome is heteromorphic in size for the long arm. Based upon the break points and the relative size of chromosome 22, four types of Ph1 chromosomes are proposed. They are: Types I (very large), II (large), III (average) and IV (small) with the break points at bands 22q13.3, 22q13.1, 22q12 and 22q11.3, respectively. The break points are arbitrary and should not be considered absolute since they are based on length differences. In two cases the Ph1 chromosome involved a translocation between chromosome 9 and 22, and the other two cases chromosome 1 or 12. Because Types I and II are hard to recognize by conventional techniques, the RFA technique (R. band by fluorescence with acridine orange) must be performed on all cases. An earlier contention that only chromosome 22 band 12 is concerned with abnormal myeloid cell proliferation in human leukaemia is rejected. Furthermore, break points are not restricted at the junction of 22q1 and q2 and 22q2 and q3 and can happen anywhere on the long arm of chromosome 22.
Haematological Reconstitution after Autografting for Chronic Granulocytic Leukaemia in Transformation: the Influence of Previous Splenectomy
Authors: Goldman John M. , Johnson Stephen A. , Islam Anwarul , Catovsky Daniel , Galton David A. G.
Published: March 2008

Summary Peripheral blood values and bone marrow appearances were monitored in eight patients treated for chronic granulocytic leukaemia in transformation by cytotoxic drugs with or without total body irradiation followed by autografting with cryopreserved‐thawed peripheral blood nucleated cells. One of the patients was ‘autografted’ on two occasions. Five patients had been splenectomized early in the first chronic phase and the other three patients had their spleens intact. Recovery of peripheral blood values was more rapid in the splenectomized than in the non‐splenectomized patients. CFUc were present in the circulation immediately after autografting in each case but subsequently the pattern of CFUc changes differed between patients. The bone marrow was hypocellular at the time of autografting but the rate at which it returned to a typical chronic phase picture varied. Peripheral blood nucleated cells collected at the time of diagnosis include stem cells with the capacity to repopulate the marrow after ‘ablative’ therapy for transformation. Elective splenectomy in the chronic phase may promote more rapid recovery of peripheral blood values but its long‐term importance is unknown.
Production of Colony Stimulating Activity in Mixed Mononuclear Cell Culture
Authors: Hellmann Andrzej , Th'ng K. H. , Goldman John M.
Published: March 2008

Summary Culture medium was harvested after co‐incubation of mononuclear cells collected and pooled from the peripheral blood of two different normal donors and was tested for colony‐stimulating activity (CSA) in agar culture. With bone marrow from normal donors or peripheral blood from patients with chronic granulocytic leukaemia as sources of granulocyte‐committed progenitor cells (CFU‐c), such mixed mononuclear cell conditioned medium (MMC‐CM) showed activity equal to that of unfractionated leucocyte feeder layers and greater than that of CSA prepared from lymphocytes stimulated by phytohaemagglutinin. The addition to plates of 2‐mercaptoethanol during the preparation of MMC‐CM enhanced CSA release. MMC‐CM is thus a convenient source of CSA and its use may be preferable to that of feeder layers when day‐to‐day reproducibility is essential.
Assessment of Complement Binding by Anti‐D and Anti‐M Antibodies Employing Labelled Antiglobulin Antibodies
Authors: Freedman J. , Massey A. , Chaplin H. , Monroe M. C.
Published: March 2008

Summary The presence of small amounts of C3d on freshly obtained normal red blood cells (RBC) was demonstrated by a radio‐labelled antiglobulin technique; this increased 1·5‐fold after incubation in fresh normal serum. No significant further increase in bound C3d was demonstrated for cDE/cDE, CDe/CDe, ‐D‐/‐D‐ RBC maximally sensitized with any of three potent anti‐D antibodies. When anti‐c, anti‐D and anti‐E antibodies were used in combination to sensitize cDE/cDE RBC, bound C3d increased by approximately 22%; however, a similar increase occurred with cde/cde RBC and was therefore considered non‐Rh specific. Umbilical cord RBC from four infants severely affected by anti‐D haemolytic disease of the newborn did not exhibit more bound C3d than cord RBC from normal controls.

One serum containing IgM anti‐M bound a small amount of complement in vitro; two IgG anti‐M sera did not.
Human Factor IX in Animals: Kinetics from Isolated, Radiolabelled Protein and Platelet Destruction following Crude Concentrate Infusions
Authors: Thompson Arthur R. , Forrey Arden W. , Gentry Patricia A. , Smith Kenneth J. , Harker Laurence A.
Published: March 2008

Summary Crude, concentrated and isolated, labelled human factor IX was prepared and infused into animals. Initially, two dogs with severe haemophilia B received both preparations and factor IX clotting activity and label survived well. Kinetic parameters fit a two‐compartment open model.

For 12 infusions of concentrates into normal baboons, clotting and antigen data again fit the model. Similar kinetics, with less variability, were found for nine infusions of [125I]factor IX. Radioactivity of the first post‐infusion samples was >90% precipitated in the double antibody system with an average of 84% of counts adsorbable to barium citrate. Compared to plasma, barium adsorbable counts gave a shorter t1/2β‐elimination phase suggesting a small recirculating 125iodide pool. When [125I]factor IX was rechromatographed over heparin‐agarose, variable amounts of altered, labelled protein were present; this species was more rapidly cleared from plasma post‐infusion.

Twelve infusions of crude concentrates into baboons were given to animals whose platelets had been labelled with 51Cr. Platelet counts as well as survival were then followed. For seven of these animals, mean counts decreased and the labelled platelets were destroyed, to an average of 21%, during the first 4 h post‐infusion. Subsequent platelet survival was normal. Three infusions of gel‐filtered, crude concentrates and two infusions of ‘activated’commercial concentrates did not significantly alter platelet survival.
An Extrinsic Factor Controls Neutrophil Alkaline Phosphatase Synthesis in Chronic Granulocytic Leukaemia
Authors: Rustin Gordon J. S. , Goldman John M. , McCarthy Donald , Mees Steven , Peters Timothy J.
Published: March 2008

Summary The mechanism underlying the reduced neutrophil alkaline phosphatase (NAP) activity in chronic granulocytic leukaemia (CGL) has been investigated by in vivo manipulation of CGL neutrophils. These cells, which had almost no alkaline phosphatase activity following collection by leucapheresis, had greatly elevated activities 14–17 h after transfusion into three neutropenic recipients. It is postulated that NAP synthesis in mature neutrophils is controlled by an extrinsic factor and that the levels of this factor is reduced in patients with CGL with high neutrophil counts.
Electron Microscope and High Resolution Autoradiographic Studies of the Erythroblasts in Haemoglobin H Disease
Authors: Wickramasinghe S. N. , Hughes Madeleine , Hollán Susan R. , Horányi Margit , Szelényi Judith
Published: March 2008

Summary Electron microscope studies of bone marrow from a patient with haemoglobin H (HbH) disease have revealed the presence of highly condensed branching intracytoplasmic inclusions within a proportion of the early and late polychromatic erythroblasts and reticulocytes. Intraerythroblastic inclusions of this type have not yet been described in any other haematological disorder and may well be a specific feature of HbH disease. Electron microscope autoradiographic studies of the marrow cells from the patient with HbH disease revealed that (1) active protein synthesis continues in inclusion‐containing erythroblasts and (2) there is a rapid precipitation of a considerable proportion of the newly synthesized free β chains onto the surfaces of preformed inclusions, presumably after conversion to HbH. These electron microscope and electron microscope autoradiographic findings are quite different from the findings previously reported in homozygous β thalassaemia using the same techniques.
Measurement of Erythropoiesis using 52Fe
Authors: Ferrant A. , Cauwe F. , Cogneau M. , Beckers C. , Verwilghen R. , Sokal G.
Published: March 2008

Summary A method for determining erythropoiesis quantitatively with 52Fe has been applied to 25 patients with anaemia. The data were derived from quantitative scanning of the erythropoietic areas and the measurement of the plasma iron turnover. We have assessed this 52Fe measurement of erythropoiesis by comparing it with total marrow iron turnover studies. Both methods give similar estimates of erythropoiesis except in patients with severe ineffective erythropoiesis and in a patient with significant peripheral haemolysis, when erythropoiesis as measured with the 52Fe technique gave higher results. In these conditions, the estimate of total erythropoiesis might be more reliable using the 52Fe quantitative scanning technique. However, the measurement of erythropoiesis with 52Fe does not distinguish effective from ineffective erythropoiesis and cannot be applied to patients with extramedullary erythropoiesis.
Serum Ferritin in Ascorbic Acid Deficiency
Authors: Roeser H. P. , Halliday J. W. , Sizemore D. J. , Nikles A. , Willgoss D.
Published: March 2008

Summary Serum ferritin levels were monitored in guinea‐pigs rendered scorbutic by dietary deprivation of ascorbic acid (AA). Hepatic and splenic concentrations of AA fell rapidly during the first 8 d of deprivation, and then declined more slowly to reach approximately 10% of control values after 14 d. Normal levels of serum ferritin (geometric mean, 324 μg/1) were observed for 20 d but a significant fall occurred between days 20 and 23 of deficiency (143 γg/1; P<0·05).

Intramuscular administration of iron dextran to scorbutic animals (75 mg Fe per kg body weight) resulted in a non‐significant elevation of serum ferritin levels (geometric mean, 470 μg/1; P>0·1). In control animals, the same dose of iron dextran caused a 10‐fold rise in serum ferritin levels (3400 μg/1). A single subcutaneous dose of AA (75 mg/kg weight) given to scorbutic animals caused a significant rise in serum ferritin levels to 599 μg/1 (P<0·001) but had no effect in control animals. The same dose of AA given to iron loaded, scorbutic animals elevated serum ferritin levels from 554 μg/1 to 1297 μg/1 (P<0·025).

Thus, AA deficiency in guinea‐pigs caused serum ferritin levels to fall and blocked the rise in serum ferritin levels observed in response to acute iron loading in control animals. Adequate tissue concentrations of AA appeared to be necessary for the maintenance of a quantitative correlation between tissue iron stores and serum ferritin levels. The mechanisms by which AA influences serum ferritin levels are at present unknown.
The Association of Eosinophilia with Lymphoblastic Leukaemia or Lymphoma: a Study of Seven Patients
Authors: Catovsky D. , Bernasconi C. , Verdonck P. J. , Postma A. , Hows J. , van der Does‐van den Berg A. , Rees J. K. H. , Castelli G. , Morra E. , Galton D. A. G.
Published: March 2008

Summary Seven patients with hypereosinophilia in association with a lymphoblastic malignancy are described. The eosinophilia preceded or was present at diagnosis in all patients. Eosinophil counts fell during complete remission but rose significantly before or during relapses in five patients. Hypogranular and sometimes Pelgereosinophils were seen in five cases. Surface and enzyme markers defined the malignancy in six cases as common‐ALL (three), T‐ALL (two) and T‐lymphoblastic lymphoma (one). Although a diagnosis of eosinophilic leukaemia or acute myeloid leukaemia with cosinophil differentiation was considered in three patients, cytochemical and ultrastructural studies failed to show any evidence of myeloid differentiation in the blast cells. The bone marrow karyotype was normal in the four patients studied.

All seven patients had one or more relapses and six died 6–62 months from diagnosis. Severe complications of the hypereosinophilic syndrome developed in one patient.

As T‐lymphocytes have been shown to be involved in the induction of eosinophilia in rodents, it is suggested that the hypereosinophilia in these patients was induced by eosinopoietic stimuli produced by lymphoblasts.
Inhibition of Normal Human Granulopoiesis in Vitro by Non‐B Non‐T Lymphocytes
Authors: Morris T. C. M. , Vincent P. C. , Sutherland Ruth , Hersey P.
Published: March 2008

Summary Normal non‐adherent mononuclear cells were shown to inhibit colony formation by normal human marrow cells cultured for 7 d in semi‐solid agar. Inhibition was the same using cells from the marrow donor or from an unrelated normal subject, and was shown to be dose‐dependent over the range of 4 × 105 to 6 × 103 mononuclear cells per 1 × 105 marrow cells plated. Inhibition was not seen in 14 d cultures, and it is postulated that colony‐forming cells sensitive to lymphocyte inhibition belonged to the population known to give rise to colonies after 7 d in culture. Cell fractionation studies showed that inhibition was due to non‐B non‐T lymphocytes, purified B cells or T cells being neither inhibitory nor stimulatory. Inhibition was only shown with intact viable lymphocytes and it was not possible to extract inhibitory activity from the cells, or to produce inhibition by media conditioned by lymphocytes. The effect was apparently due to a direct action on colony‐forming cells in the marrow and was not due to inhibition of colony stimulating activity (CSA) production, or to absorption or inactivation of CSA. These results emphasize the need to include appropriate controls when looking for possible cell‐mediated inhibitors in disease states, particularly when 7 d cultures are used.
Behaviour of Myeloid Precursors in Homozygous β Thalassaemia
Authors: Gabutti V. , Miniero R. , Piga A. , Incarbone E. , Sacchetti L. , Balegno G.
Published: March 2008

Summary Maintaining a high haemoglobin level, through a high transfusion regime, is the best method for treating thalassaemia. Not much is known about the effect of this treatment on medullary or extramedullary haemopoiesis, particularly on the extent of erythropoietic inhibition and on the behaviour of myelopoiesis.

In order to analyse some aspects of the problem, we studied the myeloid stem cells (CFU‐c) in the bone marrow and in the peripheral blood of children with homozygous thalassaemia, using the agar culture technique. The number of circulating CFU‐c observed in 68 patients was higher than in normal subjects. This number was significantly increased after splenectomy.

A positive correlation was demonstrated between the number of circulating CFU‐c and the time elapsed since the last transfusion. Patients with a high Hb level displayed a marked reduction in the number of CFU‐c in their peripheral blood.

In 10 patients, before the beginning of transfusions, bone marrow CFU‐c were lower than in normal subjects; their number increased after therapy. Most circulating CFU‐c were proliferating as shown by the thymidine suicide technique.
Relationship of Clinical Staging and Lymphocyte Morphology to Survival in Chronic Lymphocytic Leukaemia
Authors: Peterson L. C. , Bloomfield C. D. , Brunning R. D.
Published: March 2008

Summary. The value of the Rai clinical staging system and lymphocyte morphology in predicting survival in chronic lymphocytic leukaemia was examined in 83 patients who had been followed for at least 5 years. Patients with less clinical evidence of disease (Stages 0 and I) had significantly longer survivals than patients with more evidence of disease (Stages II, III and IV). Patients in whom greater than 35% of the lymphocytes resembled benign atypical lymphocytes had longer survivals than those in whom most of the lymphocytes had narrow rims of cytoplasm and coarsely clumped chromatin. The survival differences in the morphologic groups were less striking than those in the clinical stages, and when the morphological groups were corrected for clinical stage, no significant differences in survival among the morphologic groups remained.
Cytochemical Study of Thymocytes and T Lymphocytes
Authors: Basso G. , Cocito M. G. , Semenzato G. , Pezzutto A. , Zanesco L.
Published: March 2008

Summary. Fetal and postnatal thymocytes and circulating T lymphocytes were evaluated for six cytochemical reactions. Acid phosphatase activity was present in a high percentage of cells in all three groups. Beta‐glucuronidase and α‐naphthyl acetate acid esterase were negative in the most immature fetal thymocytes, but become increasingly positive with T‐cell maturation. Only the circulating lymphocytes presented a high percentage of N‐acetyl beta glucosaminidase, α‐naphthyl acetate esterase and α‐naphthyl butyrate esterase positive cells. This study discusses the presence of these enzymes as proportional to different stages in T‐cell maturation, and also of certain cytochemical phenotypes characteristic of these stages.
The Erythrocyte Membrane Disturbances in Protein‐Energy Malnutrition: Nature and Mechanisms
Authors: Fondu Pierre , Mozes Nava , Neve Pierre , Sohet‐Robazza Lucienne , Mandelbaum Israël
Published: March 2008

Summary. In protein‐energy malnutrition (PEM), as observed in Kivu, the RBC have an increased ratio of surface area to volume which is demonstrated by the presence of target cells on light microscopy and cup cells with scanning electron microscopy. The osmotic fragility is decreased. These abnormalities can be attributed to the accumulation of cholesterol and phosphatidylcholine (PC) in the RBC membrane. The molar ratio of cholesterol to phospholipids is moderately increased.

Several findings suggest that the cholesterol and PC build‐up results from disturbed exchanges in these lipids between the RBC and the plasma lipoproteins. Firstly, the osmotic fragility of a patient's RBC gradually becomes normal when the cells are transfused into a healthy recipient. Secondly, the cholesterol flux between the RBC and the plasma LDL seems to be low. Thirdly, the increase in RBC PC cannot be explained by a diminished fatty acids transport between the deep RBC PC pool and the RBC phosphatidylethanolamine (PE) pool. Finally complex disturbances of the plasma lipoproteins are obvious.

It is improbable that the cholesterol and PC build‐up accounts for the premature RBC destruction which has been described in Kivu PEM. However, the observation of an increased fatty acid turnover in RBC PC and PE, as well as other data previously obtained in Kivu PEM, lead to the conclusion that membrane peroxidation may be a major cause of the shortened erythrocyte life‐span in this syndrome.
Immune Complexes in Thrombocytopenic Patients: Cause or Effect?
Authors: Trent R. J. , Clancy R. L. , Danis V. , Basten A.
Published: March 2008

Summary. Immune complexes (ICs) in the serum of 43 patients with chronic idiopathic thrombocytopenic purpura (ITP) were measured by the C1q deviation assay during the active and inactive phases of the disease. An inverse relationship between platelet count and levels of ICs was demonstrated in all but one patient. To test whether this phenomenon was specific for chronic ITP, ICs were assayed in sera from two groups of control patients with thrombocytopenia. Group 1 had thrombocytopenia due to recognized immune mechanisms while group 2 had thrombocytopenia secondary to non‐immune mechanisms. In both these groups the degree of thrombocytopenia proved to be inversely proportional to IC levels, which was similar to the pattern observed in chronic ITP. The specificity of the assay for detection of ICs was confirmed by demonstrating a positivity rate of 65% in sera of patients with systemic lupus erythematosus, a known IC disease. On analysis the ICs were shown to have molecular weights in excess of 500000 daltons and contain variable immunoglobulin classes. The findings implicate ICs in immune destruction of platelets both in chronic ITP (as has been suggested previously) and also in thrombocytopenia secondary to known immune mechanisms. In addition the association of ICs with non‐immune thrombocytopenias is consistent with the hypothesis that platelets play an important role in clearance of ICs from the circulation, thereby protecting the vascular endothelium from damage.
A Comparison of Two Methods for Determining the Sensitivity of Human Myeloid Colony‐forming Units to Cytosine Arabinoside
Authors: Preisler Harvey D. , Epstein Joshua
Published: March 2008

Summary. Studies were carried out to determine the optimal method for measuring the sensitivity of myeloid clonogenic cells to cytosine arabinoside (ara C). Bone marrow or peripheral blood cells were either exposed to different concentrations of ara C for 1 h in vitro and then after washing were plated in agar and cultured for 7 d in vitro, or were directly plated in agar containing different concentrations of the drug. The H‐TdR suicide index of the clonogenic cells was also determined. Washing the specimens under study subsequent to incubation with ara C prior to plating in agar provided the most accurate measure of the ara C sensitivity of the clonogenic cells. When carried out in conjunction with the H‐thymidine suicide index, the ‘wash’ method permitted the simultaneous determination of both the kinetic and metabolic sensitivity of these cells to ara C. Using this combined method, it was observed that the differences between the ara C sensitivity of the CFUc of different individuals resulted from differences in the proportion of clonogenic cells synthesizing DNA in the different marrow specimens.
The Morphological Classification of Acute Lymphoblastic Leukaemia: Concordance among Observers and Clinical Correlations
Authors: Bennett J. M. , Catovsky D. , Daniel M. T. , Flandrin G. , Galton D. A. G. , Gralnick H. R. , Sultan C.
Published: March 2008

Summary. The degree of concordance in the morphological classification of ALL was assessed by the FAB group after two successive reviews of 200 and 100 slides respectively. As a result, a simple scoring system for types L1 and L2 is proposed based on the following four features: (1) nuclear cytoplasmic ratio, (2) presence, prominence and frequency of nucleoli, (3) regularity of nuclear membrane outline, and (4) cell size. By this method, the overall concordance by seven observers (agreement of 7:0 or 6:1 only) increased from 63% to 84%.

A significant difference in the incidence of the ALL morphological types in children (15 years) and adults (>15 years) was found: 74% of L1 cases were children while 66% of L2 cases were adults (P<0.001). No significant differences were observed in the incidence of L3 in children and adults or between L1 and L2 according to the membrane phenotype of the blast cells. All L3 cases had B‐cell characteristics.

A better prognosis for L1 and a higher relapse rate for L2 has been found in several recent reports; the present study may facilitate the morphological analysis of ongoing clinical trials in ALL by improving the reproducibility of the FAB classification.