bluuebunny/crossref_metadata_2025 huggingface dataset:
Patterns of in vitro BFU‐E proliferation in different forms of polycythaemia and in thrombocythaemia
Authors: Eridani S. , Sawyer B. , Pearson T. C.
Published: July 2010

To investigate erythroid colony formation in polycythaemia of different types and in thrombocythaemia, a study was performed in 121 subjects, including 13 normal volunteers (N), 30 patients with Primary Proliferative Polycythaemia (PPP), 35 with Idiopathic Erythrocytosis (IE), 19 with Secondary Polycythaemia (SP), 10 with Primary Thrombocythaemia (PT) and 14 with Secondary Thrombocytosis (ST); BFU‐E colony formation from peripheral blood samples was studied in the presence of medium only, medium plus a source of burst‐promoting activity (BPA) and medium BPA, plus erythropoietin (Ep). In the presence of medium only, practically no colonies were seen in cultures from N and SP, while a variable number was observed in cultures from PPP, IE and PT. Significant differences between groups were as follows: N v PPP; IE v PPP; SP v PPP; N v PT; ST v PT. Such differences persisted in the presence of BPA, but disappeared when Ep was added to the cultures. Further experiments, plating non‐adherent mononuclear cells in a chemically defined serum‐free medium, confirmed the growth of tiny erythropoietic colonies from circulating stem cells of patients with PPP and PT, in the absence of BPA and EP. These results provide a useful criterion to differentiate PPP, as a group, from other types of polycythaemia; individual cases of IE may need further characterization.
Cytochemical demonstration of ‘platelet’ peroxidase at the light microscope level
Authors: Khalaf M. R. , Hayhoe F. G. J.
Published: July 2010

A reliable and reproducible modified method was developed for the cytochemical demonstration of the so‐called ‘platelet’ peroxidase at the light microscopy (LM) level. Interestingly, not only normal platelets and megakaryocytes showed the peroxidase activity, but also normal lymphocytes. As with the ultrastructural platelet peroxidase (PPO) method, myeloperoxidase (MPO) was also demonstrated in granulocytes. Peroxidase activity was exhibited by blast cells of acute myeloid leukaemia, including megakaryoblasts of megakaryoblastic leukaemia, by hairy cells of hairy cell leukaemia, by centroblasts of centroblastic lymphoma and by plasma cells of plasma cell leukaemia. The enzyme activity was also demonstrated in the mast cells of systemic mastocytosis. It seems that different forms of peroxidase are present in most haemic cells. Fixation and conditions of incubation are probably the determining factors for their demonstration.

The method may allow further cytochemical discrimination at the LM level between blast cell populations which are negative for MPO by standard methods.
Different molecular defects of Gγ (Aγδβ)°‐thalassaemia in Thailand
Authors: Fucharoen S. , Winichagoon P. , Chaicharoen S. , Wasi P.
Published: July 2010

DNA from members of 2 Thai families with conditions considered to be δβ‐thalassaemia were studied by using restriction endonuclease DNA mapping. The propositus in family A is a double heterozygote for β‐thalassaemia and δβ‐thalassaemia. DNA analysis reveals a deletion of the β‐globin gene cluster starting at the area between the Sac I and Eco RI sites near the 3′ end of the Gγ‐gene and extending through the Aγ‐, δ‐ and β‐genes to an unknown extent downstream. In family B, the propositus is δβ‐thalassaemia/Hb E. Deletion of the β‐globin gene cluster begins in the large intervening sequence of the Aγ‐gene and removes both δ‐ and β‐genes downstream.
S protein/vitronectin in chronic liver diseases: correlations with serum cholinesterase, coagulation factor X and complement component C3
Authors: Kemkes‐Matthes B. , Preissner K. T. , Langenscheidt F. , Matthes K. J. , Müller‐Berghaus G.
Published: July 2010

S protein/vitronectin plays an important role as a regulatory component in the terminal steps of the complement‐ and coagulation cascades. In patients suffering from chronic liver diseases, plasma S protein concentration was measured and compared with changes in serum cholinesterase activity, coagulation factor X activity and complement component C3 concentration. Significant decreases of all these proteins were seen in liver cirrhosis. Changes in S protein concentration correlated closely with those of cholinesterase, factor X and complement C3. The data give support for the liver as the main organ of plasma S protein/vitronectin synthesis.
Characterization of a relationship between the T‐lymphocyte derived differentiation inducing factor (DIF) and lymphotoxin: A common receptor system for DIF, lymphotoxin and tumor necrosis factor downregulated by phorbol diesters
Authors: Gullberg Urban , Lantz Mikael , Nilsson Eva , Peetre Christina , Adolf Gunter , Olsson Inge
Published: July 2010

Here we describe results which show that recombinant lymphotoxin (rLT), like the T‐lymphocyte derived differentiation inducing factor (DIF), inhibited the clonogenic growth of some myeloid leukemia cell lines by concentrations of 1 to 30 pmol/l. Wild type HL‐60 cells were resistant at these concentrations but responded with differentiation into monocyte‐like cells at higher concentrations. An antigenic relationship between DIF and LT was indicated because a neutralizing monoclonal anti‐LT anti‐body bound to and neutralized both differentiation and growth inhibitory effects of DIF. An activity, which cochromatographed with DIF during all purification steps, competed with binding of both rLT and recombinant tumor necrosis factor (rTNF) to HL‐60 cells. By use of radioiodinated ligand, 2100 binding sites for rLT were detected on HL‐60 cells with a Kd of 330 pmol/l. At 37°C bound ligand was transferred to lysosomes, followed by degradation. rTNF and rLT were shown to compete for binding sites on HL‐60 cells. Receptors for both rLT and rTNF were downregulated by activators of protein kinase C such as phorbol diester or diacylglycerol; the number of cell surface receptors decreased while the Kd remained unchanged. Our observations demonstrate a functional and antigenic relationship between DIF and LT and indicate that TNF, LT and DIF share binding sites on myeloid leukemia cells that are downregulated by activation of protein kinase‐C.
Immunological reclassification of 22 children with a former diagnosis of non‐T, non‐B ALL
Authors: Fugger Lars , Platz Per , Ryder Lars , Svejgaard Arne , Heldrup Jesper , Hertz Henrik , Yssing Minna
Published: July 2010

Stored peripheral blood or bone marrow mononuclear cells from 22 pediatric patients with verified acute lymphoblastic leukemia (ALL) previously classified as non‐T, non‐B ALL were re‐investigated by flow cytometric analysis by means of a panel of B cell‐specific and ‐associated monoclonal antibodies (moabs) using a new analytical method described by Platz et al, the so‐called Delta Channel Value method. All 22 patients were immunologically re‐characterized as pre‐B ALL. The reproducibility between the first (acute) and subsequent re‐analysis was almost complete. 20 of the tumor cell populations could be assigned to the B cell differentiation scheme recently proposed by Nadler et al. This scheme operates with four stages of pre‐B cell differentiation and each stage is defined by the expression of one to four of the following markers: HLA‐DR, CD19, CD10 and CD20. Two additional markers, CD24 and CD22, were investigated in our study and allowed further subdivision of the four subgroups proposed by Nadler et al. The composition of a panel of moabs for routine classification of pre‐B ALL is proposed.
Blast crisis supervening on chronic lymphocytic leukaemia. A monoclonal progression of the disease as defined by cell surface markers
Authors: Ifrah N. , Renier G. , Saint‐Andre J. P. , Gardais J. , Hurez D. , Boasson M.
Published: July 2010

Acute leukaemia is a rare event during the course of chronic lymphocytic leukaemia (CLL), and only a small fraction of such cases have been shown to be true acute lymphoblastic crises. 1 case is described where both small lymphocytes and proliferating lymphoblasts have the same monoclonal pattern as defined by direct immunofluorescence of membrane‐bound immunoglobulins. Previous cases are reviewed and do not appear to be mere coincidence: acute blast crisis may represent a part of the natural history of CLL.
Restoration of active haemopoiesis in a patient with myelofibrosis and subsequent termination in acute myeloblastic leukaemia: Case report and review of the literature
Authors: Pamphilon D. H. , Creamer P. , Keeling D. H. , Prentice A. G.
Published: July 2010

A patient with polycythaemia vera developed typical myelofibrosis after 15 yr. After a further 8 months, during which time she was pancytopenic and transfusion‐dependent, a slow spontaneous recovery in haemopoiesis occurred and the full blood count became normal. 6 months later pancytopenia recurred and soon afterwards the patient developed acute myeloblastic leukaemia from which she died. The evolution of the later stages in this patient's illness have been documented by means of bone marrow morphology and isotopic studies. Only 2 previous reports of this kind of transformation exist in the literature, although restoration of normal or polycythaemic haemopoiesis has been reported in 8 patients with myelofibrosis. It is likely that these transformations occur because of alterations in stem cell behaviour rather than as a result of therapy.
Platelet reactivity, fibrinogen and smoking
Authors: Dotevall Annika , Kutti Jack , Teger‐Nilsson Ann‐Catrine , Wadenvik Hans , Wilhelmsen Lars
Published: July 2010

40 young healthy male volunteers (20 habitual smokers and 20 non‐smokers) were investigated with respect to platelet reactivity, plasma fibrinogen and coagulation factor VIII. Smokers had significantly lower systolic blood pressures and higher venous platelet counts. The results for ADP‐induced platelet aggregation, plasma concentrations for the 2 α‐granule proteins, beta‐thromboglobulin and platelet factor 4, did not differ between the 2 study groups involved; nor was there any difference between serum thromboxane B2 formation or plasma factor VIII:C activity. However, as compared to non‐smokers, plasma fibrinogen levels were significantly higher among the smokers.
In vitro and in vivo behavior of 111In‐labelled platelets: An experimental study of healthy male volunteers
Authors: Wadenvik Hans , Jacobsson Stefan , Kutti Jack , Syrjälä Martti
Published: July 2010

The aim of this study was to obtain a critical evaluation of a simple method for labelling platelets with 111In‐oxine. All experiments were carried out on healthy volunteers. 65 ± 7 (SD) % of the platelets in collected blood were labelled and reinjected. As compared to control experiments, only in response to a low final ADP concentration (1.0 μmol/l) did 111In‐labelled platelets show reduced in vitro aggregability. The mean platelet volume for 111In‐labelled platelets was slightly lower than the mean platelet volume in whole blood. The results for initial platelet recovery and platelet mean lifespan closely agreed with those of other studies in which considerably higher platelet extraction from whole blood was obtained. After injection, the splenic uptake and blood disappearance of 111In‐labelled platelets followed a monoexponential function with almost identical rate constants. By compartmental analysis of the equilibration of platelets between blood and spleen, the splenic blood flow was estimated to be 4.8 ± 1.9 (SD) % of the total blood volume/min; the intrasplenic platelet transit time was 9.7 ± 1.6 (SD) min, and the exchangeable splenic platelet pool 31 ± 8 (SD) %. Highly significant relationships were present between the splenic blood flow and the splenic platelet pool size, as well as between the splenic blood flow and the initial platelet recovery. It is concluded that the requirements for adequate interpretation of platelet kinetics are well met with the present method for harvesting and labelling of platelets.
A benign course of multicentric Castleman's disease with involvement of the spleen and bone marrow
Authors: Levo Yoram , Behar Albert J. , Blum Ilana , Frish Bertha
Published: July 2010

The multicentric variant of Castleman's disease (MCCD) is associated with a rapidly progressive and fatal course. The case described herein manifested unique clinical and histological features. Initial presentation as isolated splenomegaly was subsequently followed by widespread organ involvement, including lymph nodes and bone marrow. In spite of this, the patient had a very benign course of her disease. The case serves to expand even further the already wide clinical spectrum of Castleman's disease.
A tumor necrosis factor binding protein is present in human biological fluids
Authors: Peetre Christina , Thysell Hans , Grubb Anders , Olsson Inge
Published: July 2010

Tumor necrosis factor (TNF) possesses both beneficial and toxic bioactivities. Mechanisms may operate to counteract harmful effects. We have identified a TNF binding protein (TNF‐BP), which shows increased levels in serum and urine of patients on regular hemodialysis treatment (RDT). TNF‐BP inhibited the specific binding of human recombinant TNF (rTNF) to its cell surface receptor. Results from gel chromatography demonstrated the presence in serum and urine of a macromolecule with an apparent molecular weight of 50000, which formed a complex with rTNF. A 62‐fold purification of TNF‐BP from urine of patients on RDT was achieved by ion exchange chromatography and gel chromatography. Partially purified TNF‐BP reduced the growth inhibitory effect of rTNF on a susceptible leukemia cell line. TNF‐BP may act as a regulator of the biological activity of TNF and could have beneficial effects in certain inflammatory conditions.
Application of megakaryocytic morphology in diagnosing 5q‐ syndrome
Authors: Thiede Torbens , Engquist Leif , Billström Rolf
Published: July 2010

In order to evaluate the diagnostic importance of the megakaryocytic morphology in the 5q‐ syndrome we studied the bone marrow from 48 unselected patients with myelodysplastic syndromes (MDS). 44 cases were primary and 4 secondary to cytostatic drug treatment or irradiation. There were 24 cases with chromosome anomalies, of whom 10 had del (5q). 4 of these had refractory anaemia (RA) with 5q‐ as the sole anomaly (group A), 2 had RA with 5q‐ and additional chromosome anomalies consisting of trisomy 8 (group B); 3 patients had RA with excess of blasts (RAEB) and complex karyotypic changes also including 5q‐ (group B). Changes of the same type were found in 1 case of multiple myeloma with secondary MDS. All 6 RA patients with 5q‐ had characteristic megakaryocytes. More than 50% of the cells had no more than 2 nuclear segments, and predominantly had a diameter of 30 μm or more. No other patient with RA showed this picture. Only 1 patient with RAEB 5q‐ in group B had the same megakaryocytic changes. We conclude that diagnosis of a 5q‐ syndrome may be strongly suspected in cases of RA with these bone marrow changes. In cases of RAEB 5q‐ group B the bone marrow examination did not reveal the same consistent changes.
Platelet volume and intraplatelet adenine nucleotides in various hematologic disorders
Authors: Nozaki Hiroyuki , Nagao Tadami , Arimori Shigeru
Published: July 2010

By recent advanced techniques, blood platelets have proved to be varied in size and metabolism in various hematologic disorders. We examined platelet volume and intraplatelet adenine nucleotides in 36 patients with various hematologic disorders in order to clarify the quantitative platelet abnormalities. Platelet volumes were smaller in patients with acute leukemia and aplastic anemia, and larger in patients with immune thrombocytopenic purpura (ITP). The amount of intraplatelet ADP was decreased and ATP/ADP ratio was increased in acute leukemia, aplastic anemia and myeloproliferative disoders (MPD), which strongly suggested the presence of storage pool deficiency in these patients. Intraplatelet ADP per volume was decreased in acute leukemia, aplastic anemia and MPD, and ATP per volume was decreased in aplastic anemia. ATP content was increased in ITP in proportion to the increased platelet volume. These parameters were examined in 36 patients with the following hematologic disorders: 7 acute leukemia treated with cytotoxic chemotherapy, 5 aplastic anemia, 3 paroxysmal nocturnal hemoglobinuria (PNH), 9 immune thrombocytopenic purpura (ITP), 5 hypersplenism and 7 myeloproliferative disorders (MPD).
VBAMDex chemotherapy in advanced multiple myeloma*
Authors: Peest D. , Schmoll H.‐J. , Schedel I. , Glück S. , Schumacher K. , Deicher H.
Published: July 2010

In a phase II study 28 patients with advanced multiple myeloma were treated with a five drug regimen consisting of vincristine, BCNU, adriamycin, melphalan and dexamethasone. 11 out of 13 patients without prior chemotherapy showed significant remissions (> 25% tumor cell mass reduction), 7 of them had more than 75% TCM reduction. Out of 15 additional patients resistant to previous chemotherapy, 13 had significant remissions, including 9 patients with > 75% TCM reduction. No tumor progression was observed in either group of patients. The median follow‐up of all patients was 12.75 months. 4 patients relapsed. Toxicity mainly related to the bone marrow was observed in 14 patients. This regimen might offer a promising alternative for the treatment of advanced multiple myeloma, but still has to be tested in a prospective randomized trial.
Chromosomes, Auer rods and prognosis in acute myeloid leukaemia
Authors: Billström R. , Nilsson P.‐G. , Mitelman F.
Published: July 2010

We have analysed survival time and complete remission (CR) rate after induction treatment including cytosine arabinoside and an anthracycline antibiotic in 94 patients with acute myeloid leukaemia, comparing the results in four different groups according to the presence or absence of Auer rods and the presence (AN/AA) or absence (NN) of abnormal cytogenetic clones at diagnosis. The finding of Auer rods was a positive prognostic factor with respect to CR rate (p < 0.02) and survival time (p < 0.02) irrespective of the cytogenetic pattern. The AN/AA pattern was associated with lower CR rate (p < 0.05) and 6‐months survival (p = 0.05) in the 49 Auer rod‐negative patients. In the 45 patients with Auer rods, no significant differences in CR rate or survival were seen between AN/AA and NN patients. We conclude that the negative prognostic impact of chromosome abnormalities in acute myeloid leukaemia might be restricted to Auer rod‐negative disease.
Purification and characterization of eosinophil cationic protein from normal human eosinophils
Authors: Peterson Christer G. B. , Jörnvall Hans , Venge Per
Published: July 2010

ECP (eosinophil cationic protein) was purified in high yield from the granules of human buffy coat eosinophils obtained from healthy individuals. The separation procedure included gel filtration on Sephadex G‐75, ion‐exchange chromatography on Bio Rex 70, and chelating chromatography on zinc‐chelate Sepharose 6B. The normal ECP is a single‐chain, highly cationic glycoprotein which separates on SDS‐polyacrylamide gel electrophoresis into at least 3 molecular weight forms, with molecular weights of 18.5, 20 and 22 kDa. A heterogeneity in charge was also observed, with the 18.5 kDa form being the most cationic one. The various molecular species of ECP exhibited antigenic identity, identical amino acid compositions, and identical aminoterminal amino acid sequences. The molecular heterogeneity was shown to be caused by differences in glycosylation of the protein.
High‐dose retinol in children with acute myelogenous leukemia in remission
Authors: Lie Sverre O. , Wathne Karl‐Olaf , Petersen Lizette B. , Slørdahl Sofie H. , Norum Kaare R.
Published: July 2010

In a single‐institution study, 23 consecutive children with acute myeloid leukemia (AML) have been treated with a protocol including doxorubicin, cytarabine and 6‐thioguanine as induction therapy, followed by four courses of high‐dose cytarabine as consolidation. Total duration of chemotherapy was 6–8 months from diagnosis. 21 out of the 23 children achieved complete remission. During remission, the children received 52 μmol (50000 I.U.) retinol as retinyl paimitate per square meter daily. 14 of the 21 children are still in their first remission with a mean observation time of 36 months. In our study retinyl ester given in doses up to 30 times the recommended daily allowances has not caused any clinical or biochemical side effect for up to 4 yr of therapy.
Effects of platelet activating factor (PAF) on human citrated whole blood
Authors: Groscurth P. , Huracek J. , Filgueira L. , von Felten A. , Rhyner K.
Published: July 2010

Effects of PAF on citrated whole blood (C‐WB) from 38 healthy donors have been studied by impedance aggregometry and by morphologic examination of blood cells using scanning and transmission electron microscopy. In the aggregometer, the C‐WB samples showed distinct differences in PAF sensitivity. C‐WB specimens from high responders (= 15 donors) displayed a dose‐related response to PAF stimulation but those from low responders (=23 donors) did not indicate an impedance alteration even after the addition of high PAF doses (≥10−6 mol/l). Morphologic studies revealed shape‐changed platelets and primary aggregates in all C‐WB samples, whereas secondary aggregates occurred only in C‐WB specimens from high responders. Monocytes and neutrophil PMNs showed typical morphologic alterations which were observed in PAF‐stimulated C‐WB samples from all donors. Both cell types appeared polarized in shape and exhibited large vacuoles in the cytoplasm after PAF activation. In addition, monocytes came into close contact with shape‐changed platelets as well as primary and secondary aggregates, whereas PMNs had no special relationship to single or aggregated platelets. In summary, our study indicates that PAF acts on different cell types in C‐WB including platelets, monocytes and PMNs. The sensitivity of platelets against PAF stimulation appears to vary between different donors and in certain cases seems to be limited to the formation of primary aggregates.
Myelodysplastic syndromes: Analysis of morphological features related to the FAB‐classification
Authors: van der Weide M. , Sizoo W. , Krefft J. , Langenhuijsen M. M. A. C.
Published: July 2010

In a retrospective study of 96 patients with a myelodysplastic syndrome, the reproducibility of the French‐American‐British (FAB) classification was determined. Morphological abnormalities in peripheral blood and bone marrow were studied. Slides were reviewed by 3 examiners. All 3 observers agreed on morphological classification in 61% of cases, pairs of 2 in 64, 76 and 76%. The final diagnosis was refractory anaemia (RA) in 7 patients, acquired sideroblastic anaemia (AISA) in 2, chronic myelomonocytic leukaemia (CMML) in 31, refractory anaemia with excess of blasts (RAEB) in 34 and RAEB in transformation (RAEB‐t) in 22. Dyserythropoiesis, dysgranulopoiesis and dysmegakaryopoiesis were found in all FAB subgroups. Dyserythropoiesis was significantly more frequently encountered in RAEB than in RAEB‐t and CMML.
Clonal rearrangement of the T‐cell receptor γ gene associated with a bizarre lymphoproliferative syndrome
Authors: Coupe Michael , Foroni Letizia , Stamp Gordon , Lovatt Doon , Barnard Maria , Krausz Thomas , Kalofonos Haralabos , Epenetos Agamemnon , Luzzatto Lucio , Cox Timothy M.
Published: July 2010

Investigations in an unusual patient with prolonged fever, fat necrosis and haemorrhagic tendency showed widespread histiocytic phagocytosis in fat and liver biopsy specimens. Lymph nodes were also infiltrated by morphologically normal T lymphocytes predominantly expressing the OKT4 antigen. Analysis of lymph node DNA using immunoglobulin and T‐cell receptor (TCR) probes showed rearrangement restricted to the γTCR gene combined with polyclonal β TCR gene rearrangements, thus providing evidence of a unique lymphoproliferative disorder of T cells. The patient displayed a striking response to cytotoxic chemotherapy.
A case of angioimmunoblastic lymphadenopathy associated with a long spontaneous remission, retrobulbar neuritis, a clonal rearrangement of the T‐cell receptor gamma chain gene and an unusual marrow infiltration
Authors: Matthews J. H. , Smith N. A. , Foroni L.
Published: July 2010

A man with angioimmunoblastic lymphadenopathy entered a spontaneous remission that lasted 18 yr, ending in retrobulbar neuritis followed by a generalized relapse. After another remission was induced by prednisolone he developed marrow infiltration with a lymphoid cell of novel surface‐marker phenotype. His lymph node cells demonstrated a clonal rearrangement of the T‐cell receptor gamma chain (TcRy) genes whereas the TcRβ genes were heterogeneously rearranged. The marrow cells did not contain the rearranged TcRγ gene. He died of pulmonary emboli 21 yr after his original presentation. We propose a model whereby the original event was the development of a lymphoid stem cell clone with no TcR gene rearrangement. Subsequently, a sub‐clone bearing the TcRγ gene rearrangement proliferated in the lymph nodes and further heterogeneous rearrangement of the TcRβ genes occurred within this subclone. Proliferation in the marrow of the original clone finally supervened.
The splenomegaly of myeloproliferative disorders: Effects on blood volume and red blood count
Authors: Zhang B. , Lewis S. M.
Published: July 2010

By means of a scintigraphic method for estimation of spleen size, the relationship was studied between spleen size and anaemia, total blood volume and red cell volume in patients with myeloproliferative disorders. There was significant correlation between spleen size and Hb, RBC and PCV in myelofibrosis. The size of the enlarged spleen did not, however, correlate with the degree of anaemia in the leukaemias. There was positive correlation between spleen size and red cell volume in these conditions. It was not possible to evaluate the severity of polycythaemia from spleen size and the degree of enlargement of the spleen does not appear to be a reliable parameter for staging and deciding on treatment in patients with PPP.
Interleukin‐1 and tumor necrosis factor production in acute non‐lymphoid leukemia
Authors: Erroi Annalaura , Specchia Giorgina , Liso Vincenzo , Colotta Francesco , Bersani Laura , Polentarutti Nadia , Zhen‐Guo Chen , Allavena Paola , Mantovani Alberto
Published: July 2010

We have investigated interleukin‐1 (IL‐1) and tumor necrosis factor (TNF) release in 20 patients with acute non‐lymphoid leukemia (ANLL) after culture with bacterial lipopolysaccharide (LPS) or in the absence of deliberate stimulation. IL‐1 and TNF were identified by appropriate bioassays inhabitable by specific antibodies. The capacity to produce IL‐1 was expressed by most ANLL cases investigated irrespective of the FAB (French, American, British) subtype. However, the M4 and M5 cases tended to be better producers of IL‐1 than M1‐M3 cases. In contrast, TNF release was only restricted to M5 leukemias (3 out of 4 cases examined). Cytokine production may therefore provide additional criteria for a functional classification of ANLL. A considerable proportion of ANLL cases (7/18 bone marrow samples and 12/20 blood samples) released appreciable quantities of IL‐1 in culture in the absence of deliberate stimulation. “Spontaneous” TNF production was also detected in 1 out of 3 MS cases. Cells were cultured under LPS‐negative conditions and polymixin B did not affect spontaneous cytokine release. Moreover, Northern blot analysis showed that freshly isolated, non‐cultured ANLL cells expressed IL‐1 β transcripts. Inasmuch as IL‐1 is responsible for hemopoietin‐1 activity and IL‐1 induces colony stimulating factor production in various cell types, the observation of IL‐1 production in ANLL suggests that this mediator may be involved in regulatory amplifying circuits of leukemic cell proliferation.
Prolonged effect of alpha‐interferon after discontinuance of treatment in chronic myelogenous leukemia patients
Authors: Alimena G. , Montefusco E. , Mancini M. , De Fabritiis P. , Cedrone M. , Sandrelli A. , Mandelli F.
Published: July 2010

14 patients out of a group of 73 affected by CML and treated with α‐2‐IFN, who had obtained hematological and cytogenetic response, were selected to be submitted to autologous BMT. In all patients the IFN treatment was interrupted to obtain an increase of the peripheral WBC necessary for its collection by means of leukapheresis. After a median time of 20.5 weeks from discontinuance of therapy, most patients are still under hematological disease control. Serially performed cell cultures have shown deeply and persistently reduced growth patterns; cytogenetic analyses have displayed Ph' mosaicism, but with a progressive increase in time of Ph' + cells. Such findings reveal that the effect of α‐2‐IFN in CML patients can be unexpectedly prolonged after discontinuance of treatment. A possible explanation might be found in the complexity of the biological mechanisms of action of IFN, thus giving this drug promising therapeutic prospects in the treatment of CML.
Thymus and T cells are not essential for rat leucopoiesis
Authors: Benestad H. B. , Hersleth I. B. , Rolstad B.
Published: July 2010

It is generally held that T lymphocytes take part in the regulation of haemopoiesis. We have examined whether, in vivo, the influence of thymus or functional T cells is dispensable for the steady‐state or accelerated formation of granulocytes (and in some experiments macrophages), utilizing a rat model. Untreated normal and athymic ‘nude’ rats had similar blood and marrow granulocyte counts and marrow proliferative activity. Bone marrow regeneration after two cytotoxic insults to the two kinds of rats gave no clues to an important regulatory role for thymic factors or T cells. Nor were such clues obtained in experiments where bone marrow cells from normal rats were cultured in vivo in diffusion chambers (DC), in either normal rats or rats undergoing a graft‐versus‐host (GvH) reaction (with supranormal serum levels of cytokines). On the other hand, when marrow cells from athymic rats were similarly cultured in DC, small but significant differences in leucopoiesis occurred between the three kinds of DC hosts: Marrow cells lacking functional T cells generated fewer proliferative granulocytes and had a lower proliferative activity when cultured in athymic than in normal hosts, whereas the proliferative granulocytes were most numerous in chambers carried by GvH rats. No differences were found for macrophages and eosinophilic granulocytes. The results indicate that thymic hormones or T cells or both can stimulate granulopoiesis. They apparently play no indispensable role in short‐term leucopoiesis, however, since their influence was weak in our experimental models. Consequently, marked effects seen in vitro in simplified cellular systems may lose some importance in the more complex in vivo setting.
Sialyltransferase activity in plasma cells of multiple myeloma
Authors: Cohen A. M. , Allalouf D. , Djaldetti M. , Weigl K. , Lehrer N. , Levinsky H.
Published: July 2010

A marked elevation of sialyltransferase activity (STA) was observed in a solid tumor of plasma cells, which had been removed from a patient with multiple myeloma (MM), as compared to normal lymphatic tissues. STA was also determined in mononuclear bone marrow cells of 10 patients with MM and found to be 12 times higher than that of bone marrow mononuclear cells from 5 patients with non‐malignant disorders (with less than 1% plasma cells in the bone marrow aspirate). A significant correlation was found between STA and the number of plasma cells in the bone marrow aspirate.
Can late relapse be predicted at initial diagnosis in childhood acute lymphoblastic leukemia?
Authors: Rautonen Jukka , Siimes Martti A.
Published: July 2010

We have investigated whether any prognostic factor can be used to identify those children who have a relapse after discontinuation of therapy for acute lymphoblastic leukemia (ALL). Our population‐based series comprised 167 children with newly diagnosed ALL. The 3‐year event‐free survival rate in these children was 65%. Maintenance therapy was electively discontinued for 120 patients, 20 of whom have subsequently had a relapse 1 to 27 months later. In multivariate analysis the risk of late relapse in the 15 patients with initially enlarged kidneys was 4.5‐fold (95% confidence limits 1.7–11.8) that of the others (p < 0.01). The risk in the 18 patients with initially elevated CSF protein concentration (> 0.4 g/l) or leukocyte count (> 5 × 106/l), but with no blasts in the CSF, was 3.8‐fold (1.5–9.6) that of the others (p < 0.01). Our results indicate that enlarged kidneys or abnormal CSF findings at initial diagnosis are associated with an increased risk of late relapse in children with ALL.
Heme‐binding plasma membrane proteins of K562 erythroleukemia cells: Adsorption to heme‐microbeads, isolation with affinity chromatography
Authors: Majuri R.
Published: July 2010

Heme‐microbeads attached themselves to the surface of viable K562 cells in a manner inhibitable by free hemin, indicating heme‐receptor interaction. The microbeads were at first evenly distributed, but after prolonged incubation at 37°C they formed a cap on one pole of the cells indicating clustering of the membrane heme receptors. Membrane proteins were labeled by culturing the cells in the presence of 35S‐methio‐nine and were then solubilized with Triton X‐114. The hydrophobic proteins contained about 20% of the total bound label. The solubilized membrane proteins were subsequently adsorbed to a heme‐Sepharose affinity gel. According to SDS‐electrophoresis and subsequent autoradiography, the immobilized heme captures two proteins or a protein with two polypeptides of 20000 and 32000 daltons. The larger of these was only weakly labeled with 35S. The same two bands were observed if the cell surface proteins were labeled with 125I by the lactoperoxidase method and the subsequently solubilized membrane proteins were isolated with heme‐Sepharose.
Primarily asymptomatic low‐grade non‐Hodgkin lymphomas: Prediction of symptom‐free survival and total survival
Authors: Martinsson Ulla , Glimelius Bengt , Hagberg Hans , Sundström Christer
Published: July 2010

Low‐grade non‐Hodgkin lymphomas (NHL) constitute a group of tumours with an often long survival time but, at present, with little — or no — chance of cure if the disease is not strictly local. In primarily asymptomatic patients, treatment may either be started immediately after diagnosis or deferred until symptoms occur. The possibility of predicting the symptom‐free time was investigated in 64 non‐selected initially asymptomatic patients with advanced low grade NHL, all of whom had treatment deferred until symptoms occurred. The most powerful predictor was the histopathological subgroup. Lymphocytic (LC) and follicular centroblastic‐centrocytic (fCBCC) lymphomas had a median symptom‐free period of 2 years, which was four times longer than that for immunocytoma (IC) and follicular and diffuse CBCC (fdCBCC). In addition, the serum levels of deoxythymidine kinase (S‐TK) and lactic dehydrogenase (S‐LDH) could predict the symptom‐free period. This did not apply to S‐Haptoglobin, S‐Orosomucoid or stage. In a multivariate analysis, only S‐TK gave additional information to histopathology. The only variable that predicted the overall survival time was the length of the symptom‐free period.
Immunological and molecular biological identification of a true case of T‐hairy cell leukaemia
Authors: Demeter Judith , Pálóczi Katalin , Földi János , Hokland Marianne , Hokland Peter , Benczúr Miklós , Lehoczky Dezsö
Published: July 2010

A hairy cell leukaemia (HCL) patient is presented in whom the peripheral blood mononuclear cells (PBMCs) carried suppressor T‐cell markers (CD3 +, CD2 +, CD8 + /CD4‐, CD38 +). Analysis of genomic DNA of PBMNC showed the presence of a monoclonal population of T cells, the T‐cell receptor (TCR) beta‐chain gene being rearranged on both alleles (DR/DR), while the immunoglobulin (Ig) heavy chain‐genes were in germline configuration. The neoplastic cells were found to react with the monoclonal antibody RAB‐1 — originally described as belonging to the B lineage‐restricted monoclonal antibodies — and to carry RAB‐1/CD‐8 in a double marker assay. Natural killer activity of PBMNCs against K562 target cells was severely reduced, while the cells were found to exert strong antibody‐dependent cellular cytotoxicity.
Clonal cell surface structures related to differentiation, activation and homing in B‐cell chronic lymphocytic leukemia and monoclonal lymphocytosis of undetermined significance
Authors: Kimby Eva , Mellstedt Håkan , Björkholm Magnus , Holm Göran
Published: July 2010

Cell surface structures related to differentiation, activation and “homing” were identified on the leukemic cell clone in blood of 64 patients with a monoclonal B‐cell lymphoproliferative disorder. Patients were selected with regard to clinical signs and symptoms of the disease. 39 patients had progressive chronic lymphocytic leukemia of B‐cell type (B‐CLL): 16 with lymph node enlargement and 14 with progressive lymphocytosis as the most prominent symptom, respectively. 1 patient had an isolated splenomegaly and 8 had symptoms from enlarged lymph nodes, lymphocytosis and/or splenomegaly. 25 patients had an isolated monoclonal B‐cell lymphocytosis in blood and bone‐marrow but no other signs or symptoms of the disease. The lymphocytosis in these patients was considered to be of “undetermined significance” and the term B‐cell lymphocytosis of undetermined significance (B‐MLUS) was used. Patients with a prominent lymphadenopathy and/or splenomegaly had CD22+ leukemic cells while in patients with a progressive lymphocytosis the B‐cell clone expressed Leu‐8. Thus, CD22 might be related to the homing capacity of B lymphocytes for lymphnodes and spleen, while Leu‐8 might define a circulating B‐cell subset. In B‐MLUS about 50% of the monoclonal B cells co‐expressed Leu8 which is consistent with a more differentiated phenotype compared to B‐CLL with progressive lymphocytosis. The CD22 expression was mostly low in B‐MLUS although a few patients showed high values. The expression of receptors for growth factors (CD23, CD25, CD71) was higher in B‐CLL compared to B‐MLUS patients (p < 0.001), which is consistent with a difference in lymphocyte activation stage and/or response to growth factors.
Chronic myelogenous leukemia and Klinefelter's syndrome
Authors: Oguma Nobuo , Takemoto Motoyoshi , Oda Kenji , Tanaka Kimio , Shigeta Chiharu , Sakatani Keiko , Kamada Nanao , Kuramoto Atsushi
Published: July 2010

A case of chronic myelogenous leukemia with Klinefelter's syndrome mosaicism in a 27‐yr‐old male is reported. Cytogenetic analysis provided evidence that the Philadelphia chromosome occurred monoclonally in the XXY cells but not in the XY cells.
Endogenous megakaryocyte colonies from peripheral blood in precursor cell cultures of patients with myeloproliferative disorders
Authors: Battegay E. J. , Thomssen C. , Nissen C. , Gudat F. , Speck B.
Published: July 2010

Megakaryocyte colony formation, as identified by conventional techniques, was observed in precursor cell cultures from peripheral blood in 8 of 20 consecutive patients with diagnosis of myeloproliferalive disease (4/11 patients with polycythemia vera, 3/5 with essential thrombocythemia, 1/2 with primary osteomyelofibrosis and 2 with a myeloproliferative syndrome not further assessable), but not in 50 healthy controls (p < 0.0001). 7 cultures showed spontaneous erythroid colonies, but were negative for megakaryocyte colonies. Megakaryocyte colony formation was independent of added erythropoietin, plasma or human leukocyte‐conditioned medium, but was dependent on the presence of accessory cells. The cells in megakaryocyte colonies had the characteristic morphology of megakaryocytes and stained positively with the IIIa/IIb monoclonal anti‐platelet antibody. Thus, megakaryocyte colony formation by precursor cells from peripheral blood in the absence of exogenous stimulating factors seems to be a phenomenon specific for myeloproliferative disease. Differential diagnosis of thrombocythemia may be facilitated by demonstration of endogenous megakaryocyte colony formation, which does not occur in secondary disease.
Fraction of S‐phase cells in blood mononuclear cells in non‐Hodgkin's lymphomas—Correlation with clinical features and prognosis
Authors: Lindh Jack , Jonsson Håkan , Lenner Per , Roos Göran
Published: July 2010

A consecutive material of 111 untreated patients with non‐Hodgkin's lymphoma was studied with respect to fraction of S‐phase cells in blood mononuclear cells in relation to presence of monoclonal B cells in blood (MBCB). Fraction of S‐phase cells was determined by flow cytometry and estimation of MBCB was performed by kap‐pa:lambda analysis. The fraction of S‐phase cells was significantly higher (p < 0.001) in MBCB‐positive cases (median 1.2%) than in the MBCB‐negative (median 0.7%). MBCB‐positive patients with S‐phase values ≥ 1.5% had a less favourable prognosis compared to those with < 1.5% cells in S‐phase (p = 0.01). In a Cox multiparameter analysis, advanced clinical stage, high‐grade morphology and high fraction of S‐phase cells in blood in MBCB‐positive cases were independent, statistically significant, negative prognostic indicators. The results indicate that an elevated S‐phase value in blood in non‐Hodgkin's lymphoma constitutes a negative prognostic factor, probably reflecting proliferating tumour cells in blood.
Thymidylate synthesis and utilization via the de novo pathway in normal and megaloblastic human bone marrow cells
Authors: Matthews J. H. , Armitage J. , Wickramasinghe S. N.
Published: July 2010

We have measured the thymidylate synthetase activity of intact bone marrow cells using a 3H2O release assay. The mean thymidylate synthetase activity of vitamin B12‐ or folate‐deficient megaloblastic marrow cells was reduced only in severely anaemic patients. There was a correlation between thymidylate synthetase activity and RBC in patients with megaloblastic haemopoiesis. The mean rate of incorporation into DNA of 6‐3H deoxyuridine was similar in megaloblastic and normoblastic marrows. The rate of thymidylate synthesis exceeded its incorporation into DNA in all marrows, and the mean ratio between synthesis and incorporation was similar in normoblastic and megaloblastic patients, being independent of both thymidylate synthetase activity and RBC. Thus de novo thymine nucleotides were not utilized more efficiently in megaloblastic marrow cells. These data suggest that impaired thymidylate synthesis may not be the central defect in megaloblastic haemopoiesis, and that there is only a single pool of thymidine triphosphate in human bone marrow cells.
Dual rearrangement of immunoglobulin and T‐cell receptor genes in blast crisis of CML
Authors: Uike Naokuni , Takeichi Naohisa , Kimura Nobuhiro , Takahira Hiroyuki , Kozuru Mitsuo
Published: July 2010

Dual rearrangement of immunoglobulin and T‐cell antigen receptor (β, δ) genes was demonstrated in a case of Philadelphia chromosome‐positive chronic myeloid leukemia (CML) in blast crisis. The blast cells, showing L2 morphology and high activity of TdT, expressed pre‐B cell (CD19+, Ia+) and myeloid (CD13+, CD34+) surface antigens but lacket T‐cell antigens (CD2‐, CD7). Cytogenetic studies on bone marrow and peripheral blood revealed the Phl chromosome in all metaphases analyzed, majority of which also had the additional chromosome changes, +8, +10, +21. Furthermore, molecular analysis of the breakpoint cluster region (bcr) on chromosome 22 showed a rearrangement, confirming the CML origin of the blast cells.
Monoclonal and biclonal immunoglobulin‐producing disorders
Authors: Österborg A. , Mellstedt H.
Published: July 2010

Monoclonal gammopathies are B lymphocyte‐derived disorders producing a specific monoclonal immunoglobulin. The group includes multiple myeloma, Waldenström's macroglobulinaemia, monoclonal gammopathy of undetermined significance (MGUS) and heavy‐chain disease. In 1% of the cases a double M‐component is present which is referred to as a biclonal gammopathy. The pathogeneses of these disorders are unknown, but several possibly related environmental factors have been identified. In multiple myeloma, an increased incidence and mortality rate has been noted during the last 20 years. Strong lines of evidence are presented showing that, in monoclonal gammopathies, the cell clone consists of B lymphocytes in different stages of maturation. In most cases of biclonal gammopathy there is probably a common cellular origin.
Biologic and clinical significance of cytokine production in B‐cell malignancies
Authors: Torcia Maria , Aldinucci Donatella , Carossino Anna Maria , Imreh Farhad , Cozzolino Federico
Published: July 2010

Cytokines are a group of polypeptide hormones endowed with pleiotropic biological properties. Normal B lymphocytes produce a number of these factors that subserve important regulatory functions in the combined processes of proliferation and differentiation. Also neoplastic B cells can release cytokines and, simultaneously, respond to the same factors in an autocrine circuit that supports their malignant growth. In addition, tumor cells can make use of the factors released by normal cells, either spontaneously or under the influence of inductive signals from the neoplastic cells. Inappropriate or excessive release of cytokines may have an important role in the pathophysiology of some clinical features. Thus, neutralization of cytokine biologic activity in vivo could be a therapeutic strategy for treatment of human B‐cell neoplasias.
High‐dose chemotherapy and autologous bone marrow transplantation for myeloma
Authors: McElwain T. J. , Selby P. J. , Gore M. E. , Viner C. , Meldrum M. , Millar B. C. , Malpas J. S.
Published: July 2010

In three studies we have attempted to increase the complete remission rate in a series of patients with multiple myeloma under the age of 69. A CR rate of 27% was seen in 63 patients treated with intravenous high‐dose melphalan 140 m/m2 with or without the addition of high‐dose methyl prednisolone ***1g/m2 for 5 days (in 22 patients). In a third study a CR rate of 50% was seen in 50 patients treated in a programme in which vincristine, adriamycin and methyl prednisolone was first given and patients then received high‐dose melphalan 140–200 mg/m2 with an autologous bone marrow transplant where possible. Median remission duration in the first two studies was 19 months with a median survival of 5 years. A definition of complete remission in myeloma is proposed.
Ex vivo treatment of myeloma cells by 4‐HC, VP‐16, LAK cells and antibodies
Authors: Gulati Subhash C. , Shimazaki Chihiro , Lemoli Roberto M. , Atzpodien Jens , Clarkson Bayard D.
Published: July 2010

To assess the usefulness of 4‐hydroperoxycyclophosphamide (4‐HC) and Etoposide (VP‐16) as a purging agent for myeloma cells in bone marrow ex‐vivo, myeloma cell lines (SK‐RCS‐1, RPMI‐8226), lymphoma cell line (SK‐DHL‐2) and normal bone marrow (BM) cells were treated at different concentrations of 4‐HC, VP‐16. In separate experiments, LAK cells or antibodies were also used to treat the above cell lines. Clonogenic tumor cells from all three cell lines could be reduced by more than 4 logs, when treated alone or as a mixture with irradiated normal bone marrow cells at a 4‐HC concentration of 60 μmol/l. Under similar conditions, approximately 1% of normal BM myeloid progenitor granulocyte‐macrophage colony forming cells (CFU‐GM) survived. The results with LAK cells and antibodies were also encouraging. These observations support the use of various purging methods for myeloma cells for autologous bone marrow transplantation.