bluuebunny/crossref_metadata_2025 huggingface dataset:
Bone marrow culture and haemopoietic growth factors: Recent developments and current status
Authors: Mortensen Børge Thing , Knudtzon Søren , Hovgaard Doris , Jensen Asbjørn Nymark
Published: July 2010

The use of semi‐solid cultures of bone marrow cells is briefly reviewed. In the last two decades this technique has greatly improved our understanding of the hierarchical structure of cell production and its regulatory factors. It is now well established that relatively few multipotent stem cells, with self‐renewing capacity, are feeding cells into the differentiating pool of all the different lineages of mature cells found in blood. The proliferation and differentiation in this system are controlled by specific haemopoeitic growth factors, which also regulate the function of the mature cells. Several of these factors are now produced in recombinant form, allowing further studies of their biological significance. Clinical trials with two of the factors have supported the experimental evidence for their in vivo function, and they can be administered without serious side effects. The clinical access to these factors is promising for future treatment of cancer patients and may possibly completely change the outcome of several haematological and infectious diseases.
Behenoyl cytosine arabinoside, aclacinomycin A, 6‐mercaptopurine, and prednisolone combination therapy for acute non‐lymphocytic leukaemia in adults
Authors: Yoshida Minoru , Suda Keiichi , Tsuboyama Akihiro , Sasaki Ryuhei , Kitagawa Seiichi , Kano Yasuhiko , Muto Yoshitomo , Takaku Fumimaro , Sakamoto Shinobu , Miura Yasusada
Published: July 2010

38 consecutive, previously untreated adult patients with acute non‐lymphocytic leukaemia (ANLL) were treated with BHAC‐AMP (N4‐behenoyl‐1‐β‐D‐arabinofuranosyl‐cytosine, aclacinomycin A, 6‐mercaptopurine, and prednisolone) therapy between March 1980 and February 1985. 25 patients (65.8%) achieved complete remission (CR). Median CR duration and median survival of patients who achieved CR were 14, and 24 months, respectively. The Kaplan‐Meier analysis revealed a probability for remaining in CR of 18.0% at 5 years. Analysis of failure cases revealed that most of them were due to resistant disease. Major toxicities were infection, diarrhoea, liver dysfunction, nausea and vomiting but these were acceptable. The results indicate that BHAC‐AMP therapy is comparable to the regimen with daunorubicin and cytosine arabinoside and a further clinical trial is necessary for previously untreated adult patients with ANNL.
In vitro and in vivo characterization of a high‐purity, solvent/detergent‐treated factor VIII concentrate: Evidence for its therapeutic efficacy in von Willebrand's disease
Authors: Mazurier Claudine , De Romeuf Christophe , Parquet‐Gernez Armelle , Goudemand Maurice
Published: July 2010

A factor VIII (FVIII) concentrate, virus‐inactivated by the solvent/detergent procedure, was studied in vitro. In contrast with most high‐purity, virus‐inactivated FVIII concentrates, it contains not only high levels of von Willebrand factor (vWF) antigen and ristocetin cofactor activity but also high molecular weight forms of von Willebrand factor. Furthermore, it is able to promote platelet adhesion on collagen in a perfusion system. In vivo studies performed in patients with different types of von Willebrand's disease provided evidence that this concentrate corrects Duke's bleeding time and prevents or stops haemorrhages. Thus, the particular advantages of this FVIII/vWF preparation are safety, low content of contamination proteins, and efficacy in von Willebrand's disease.
Prognostic significance of peripheral blood S + G2/M phase size in adult acute non‐lymphoblastic leukaemia
Authors: Guerci A. , Pierrez J. , Kohler F. , Guerci O.
Published: July 2010

Flow cytometric analysis of peripheral blood (PB) S + G2/M phase size was performed in 73 audit patients with untreated acute non‐lymphoblastic leukaemia, to assess whether the results may correlate to response rate and patient prognosis. All patients were treated with the same induction chemotherapy regimen: ARA‐C alone or in combination with an anthracycline antibiotic. Pretreatment PB S + G2/M phase size is significantly correlated to induction response rate (p < 0.02), duration of response (p<0.02) and duration of survival. Patients with low PB S + G2/M phase size experience a longer survival, in patients over and below 50 yr (p < 0.001). Lastly, early deaths tend to be more frequent in the high median age and high PB S + G2/M phase size group. Our study suggests that PB S + G2/M phase size has prognostic significance in obtaining response and duration of survival.
The disturbance of hemostasis in septic shock: role of neutrophil elastase and thrombin, effects of antithrombin III and plasma substitution*
Authors: Seitz R. , Wolf M. , Egbring R. , Havemann K.
Published: July 2010

In 42 patients with septic shock, 29 of whom underwent substitution with antithrombin III concentrate and fresh frozen plasma for coagulation disorders, the proteinase‐inhibitor complexes thrombin‐antithrombin III and neutrophil elastase‐alpha1 proteinase inhibitor, were elevated on admission. On admission, the elastase complex was significantly higher in the patients receiving substitution (p = 0.0039), but at the endpoint it was higher in the non‐survivors (p = 0.0040). The elastase decrease was confined to the substitution group with the thrombin complex decreasing in both groups. Initially the thrombin complex correlated with prothrombin times and factor XIII, while the elastase complex correlated with creatinine, thrombocyte count and prothrombin times in the late stages. Hemostatic disturbance, thrombin generation and neutrophil elastase release were favorably influenced by substitution. Furthermore, in this uncontrolled pilot study, the survival rate was higher in the treated (16 of 29) than in the untreated (1 of 13) patients, although the treated patients initially had pronounced hemostatic disturbances.
Antibodies to platelet membrane glycoprotein antigens in three cases of infectious mononucleosis‐induced thrombocytopenic purpura
Authors: Winiarski Jacek
Published: July 2010

Infectious mononucleosis may occasionally be complicated by purpura, but the mechanism of the thrombocytopenia is not known in detail. In the present study, 3 children with mononucleosis‐associated purpura were found to have marked elevations of platelet‐associated immunoglobulins and circulating platelet binding IgG and IgM. Employing electrophoretically (SDS‐PAGE) separated normal platelet membrane proteins in an immunoblot assay, serum IgG and IgM antibodies were found to be directed to antigenic determinants situated on glycoprotein GP IIb (140 kDa) in all patients, but also on smaller proteins of molecular weights between 30 and 52 kDa. 24 control sera were negative. Positive reactions were eliminated after absorption of sera with fresh platelets. These results demonstrate autoantibodies to platelet surface membrane proteins in infectious mononucleosis‐induced purpura and suggest a transient autoantibody‐mediated platelet destruction as the cause of thrombocytopenia in these patients.
Immunoelectron microscopic demonstration of antigenic sites on lymphoid cells using a human monoclonal antibody (Ha6D3)*
Authors: Hansmann M. L. , Harpprecht J. , Westphal E. , Müller‐Ruchholtz W.
Published: July 2010

The reactivity of a human monoclonal antibody directed against human B and T lymphocytes was tested for the first time at the ultrastructural level. The antigenic sites detected by this antibody were localized on the surface membrane of lymphocytes and, to a lesser extent, in the cytoplasm on membranes of the endoplasmic reticulum and perinuclear envelop of some centroblasts and immunoblasts. Ultrastructural demonstration of target antigen detected by human monoclonal antibodies may be important prior to therapeutic application of these antibodies.
Preferential localisation of red cell vacuoles to pathologically shaped human red blood cells
Authors: Demeter Judith , Magyar Zoltan , Varga Fatima , Szathmári Miklós , Lehoczky Dezsö , Ellegaard Jørgen
Published: July 2010

The percentage of pitted erythrocytes and Howell‐Jolly bodies in peripheral blood samples of 51 individuals following posttraumatic splenectomy and 20 patients splenectomized because of various haematological diseases differed significantly from each other (p < 0.001) and from that of healthy controls (p < 0.001). The percentage of pitted erythrocytes was significantly higher in pathologically shaped red blood cells (RBCs) (acanthocytes, schizocytes, elliptocytes) than in normal discoid shaped RBCs (p < 0.001). As the number of pits per RBC showed great individual variations, a scoring system for the evaluation of pitted RBCs is proposed.
Antibody response to pneumococcal vaccine in patients with early stage Hodgkin's disease
Authors: Frederiksen Birgitte , Specht Lena , Henrichsen Jørgen , Pedersen Freddy Karup , Pedersen‐Bjergaard Jens
Published: July 2010

Antibody response to pneumococcal vaccination was studied in 76 patients with Hodgkin's disease (HD) before, during and at different time intervals after cessation of therapy. All patients were in pathological stage I and II following explorative laparatomy with splenectomy. The increase in antibody response was compared to the findings in 12 healthy volunteers with the aim of establishing the optimal time for vaccination. Serum antibodies against 6 of the pneumococcal polysaccharide antigens (types 1, 4, 7F, 14, 18C and 23F) contained in the vaccine were determined by an ELISA. Antibody response to pneumococcal type antigens was similar in healthy adults and in patients with early stage HD before therapy. After treatment, post‐vaccination antibody response became negligible. Even up to 7 years after cessation of therapy patients were not able to raise a significant antibody response.
Study of HLA antigens in patients with Hodgkin's disease
Authors: Pražák J. , Heřmanská Z.
Published: July 2010

This paper studies relations of 21 HLA antigens of the A and B loci to selected characteristics in a group of patients with Hodgkin's disease. The investigated characteristics are age, sex, histological patterns, clinical stage and symptomatology. The considered group consists of 68 patients, 37 males and 31 females aged from 19 to 66 years and from 26 to 63 yr, respectively. A comparison between antigen frequencies for patients and controls (301 unrelated blood donors) is carried out. The results confirm that there is no relation between HLA antigens and Hodgkin's disease. A significant increase of All is found for females under 40 yr. Certain HLA antigens show significant correlation neither with the patient age nor with clinical or histological stages. A significant increase of B5 is observed for 16 relapsed patients. For them, significant test characteristics are found in the presence of cross‐reacting HLA‐B5, B35.
Alternating combination chemotherapy (VMCP/VBAP) is not superior to melphalan/prednisone in the treatment of multiple myeloma patients stage III —A randomized study from MGCS
Authors: Österborg A. , Ahre A. , Björkholm M. , Björeman M. , Brenning G. , Gahrton G. , Gyllenhammar H. , Johansson B. , Juliusson G. , Järnmark M. , Killander A. , Kimby E. , Lerner R. , Nilsson B. , Paul C. , Simonsson B. , Stalfelt A.‐M. , Strander H. , Smedmyr B. , Svedmyr E. , Udén A.‐M. , Wadman B. , Wedelin C. , Mellstedt H.
Published: July 2010

86 previously untreated patients with multiple myeloma stage III entered a randomized trial comparing combination chemotherapy (VMCP/VBAP) (n = 42) with intermittent oral melphalan and prednisone (MP) treatment (n = 44). The treatment gropus were well comparable with regard to major prognostic factors. There was no statistically significant difference in the response rates, 52% (VMCP/VBAP) vs 61% (MP); in the response duration times, median 19 months vs 22 months, or in the survival times, median 24 months vs 28 months. However, survival of patients older than 65 years was significantly shorter in the VMCP/VBAP group (median 15 months) compared to the MP group (median 23 months) (p = 0.03). No significant difference in non‐hematological or hematological toxicity was noted. The study further supports the notion that MP therapy should be used as primary standard treatment for patients with multiple myeloma.
The splenomegaly of myeloproliferative and lymphoproliferative disorders: Splenic cellularity and vascularity
Authors: Zhang B. , Lewis S. M.
Published: July 2010

Employing radionuclide scanning, the volume of the spleen, its red cell pool and plasma pool have been measured in vivo, and the relative proportions of cellularity and vascularity of the spleen have been calculated in 51 patients with myeloproliferative and lymphoproliferative disorders. In primary proliferative polycythaemia (polycythaemia vera), the increase of spleen size was attributed mainly to the increase of splenic vascularity; in myelofibrosis and in hairy cell leukaemia, the increase of spleen size was associated with increase in both splenic vascularity and cellularity, whilst in CGL and CLL the increase was attributed more to cellularity than to vascularity.
Dual granule localization of the dormant NADPH oxidase and cytochrome b559 in human neutrophils
Authors: Bjerrum O. W. , Borregaard N.
Published: July 2010

The subcellular localization of the microbicidal nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and associated b‐cytochrome was investigated in human neutrophils. In unperturbed neutrophils 85% of b‐cytochrome and the major part of membrane‐bound components of the NADPH oxidase co‐sedimented with markers for specific granules and gelatinase. Using cytochrome b559 as a marker for membrane‐bound components of the NADPH oxidase in quantitative studies we observed that, of the remaining 15%, the vast majority co‐sedimented with latent alkaline phosphatase, a marker for a newly identified mobilizable intracellular compartment. Only a small fraction co‐localized with the plasma membranes. Azurophil granules contained a protease activity which rapidly inactivated the NADPH oxidase components present in other membranes. Stimulation of the neutrophils with formyl‐methionyl‐leucyl‐phenyl‐alanine and leukotriene B4 which caused minimal degranulation of specific granules, resulted in translocation of b‐cytochrome to the plasma membrane, concomitant with incorporation of alkaline phosphatase into the plasma membrane.
Chemical and immunological characteristics of four different L‐asparaginase preparations
Authors: Koerholz D. , Brueck M. , Nuernberger W. , Juergens H. , Goebel U. , Wahn V.
Published: July 2010

We studied the differences in protein composition and immunologic reactivity of two E. coli‐derived L‐asparaginase (1‐Asp) preparations (I and II), Erwinia‐Asp (III) and PEG‐modified E. coli 1‐Asp (IV). On gel filtration, each of preparations I‐III showed three major peaks at 100, 270 and 460 KD, all with enzyme activity, whereas PEG‐Asp showed peaks at 35 and 220 KD. On SDS‐PAGE one major subunit could be identified at 32 KD (I and II) or 40 KD (III), whereas PEG‐modified 1‐Asp could only be detected by lowering the polyacrylamide concentration and gave a single band above 200 KD. Using a polyclonal rabbit antibody generated against preparation I, only the E. coli 1‐Asp preparations (I and II) formed precipitin lines on Ouchterlony double diffusion. After freezing and thawing, preparation IV also reacted with this antibody. In sera from patients treated with preparation I, antibodies (detected by ELISA) reacted with preparations I and II but not with preparations III and IV. These results indicate that Erwinia‐Asp (III) and PEG‐Asp (IV) are distinct from E. coli preparations (I and II) by molecular weight and immunological behavior. They also provide an experimental rationale for the use of Erwinia‐Asp as well as PEG‐Asp in E. coli Asp‐sensitized patients.
Lymphokine‐activated killer (LAK) cells inhibit the clonogenic growth of human leukemic stem cells
Authors: Lista Patrizia , Fierro Maria Teresa , Liao Xin‐Sheng , Bonferroni Margherita , Brizzi Maria Felice , Porcu Pierluigi , Pegoraro Luigi , Foa Robert
Published: July 2010

The effect of lymphokine‐activated killer (LAK) cells on the in vitro clonogenic capacity of acute myeloid leukemia (AML) blasts was investigated in a semisolid medium assay. The leukemic clonogenic capacity of 11 AML cases, selected on the basis of their ability to grow in vitro, was highly reduced following overnight preincubation with LAK effectors. The degree of colony inhibition, which ranged between 66% and 98% (mean 83.8% ± 11.4 SD), was quantitatively greater than by 51Cr release, which gave rise to lytic values between 5% and 65% (mean 43.2% ± 19.2 SD). The demonstration that the clonogenic inhibition was still induced following a shorter pre‐incubation period (4 hours) suggests that the effect is unlikely to be due only to the generation of cytotoxic activity during the incubation time. The possibility that LAK cells may be employed in the management of residual disease is strengthened by the evidence that the clonogenic potential of samples containing as few as 20% and 14.3% leukemic cells could be almost completely abolished by LAK effectors. These findings further point the possible role of adoptive immunotherapy with interleukin 2/LAK cells in the treatment of patients with acute leukemia.
High‐dose intravenous methylprednisolone for acute childhood idiopathic thrombocytopenic purpura
Authors: Özsoylu Ş. , Irken G. , Karabent A.
Published: July 2010

49 children with acute idiopathic thrombocytopenic purpura (ITP) were divided into non‐treatment, oral prednisone (2 mg/kg), and high‐dose intravenous methylprednisolone (HIVMP) treatment groups which consisted of 17, 16 and 16 children respectively. Platelet counts rose above 150000/μl over a 2‐week period in 5 (29.4%) children in the first group, 5 (31.2%) in the second group and 15 (93.7%) children in the third group. Platelet counts reached the normal level in only 3 days in 11 (68.7%) children treated with HIVMP. Initially, antiplatelet antibodies (APA) were shown by the Handin and Stossel method in every patient. With normalization of platelet counts, the antibodies decreased but could still be detected in every case; antibody decrease was greater in the HIVMP group. With the exception of mild cushingoid appearance, none of the major corticosteroid side effects was observed in the treated children.
Differential expression of CD25 and HC2 antigens on subtypes of acute myeloid leukemias
Authors: Padrós María R. , Salamone Maria C. , Zunszain Patricia A. , Fainboim Leonardo
Published: July 2010

The presence of CD25 and HC2 antigens in 66 different patients with acute myeloid leukemia (AML) was investigated. The expression of both antigens was observed in 32% of AML cells. Dual fluorescence staining experiments performed in 5 AML patient cells showed that CD25 and HC2 antigens were simultaneously expressed in a single cell. The expression of both antigens was mainly observed in the M4 and M5 subtypes of AML. Although immunopurified IL‐2 was able to block the binding of anti‐CD25 monoclonal antibody to the AML cells, the IL‐2 receptor did not appear to be functional.
Acute myelogenous leukemia in first relapse treated with two consecutive autologous bone marrow transplantations: A pilot study
Authors: Meloni Giovanna , De Fabritiis Paolo , Pulsoni Alessandro , Sandrelli Antonella , Simone Francesca , Mandelli Franco
Published: July 2010

The feasibility and the antileukemic activity of a double sequential autograft has been evaluated in 3 patients affected by AML in first hematological relapse. Bone marrow collection and cryopreservation was performed twice: during first complete remission (CR) and during second CR. At the time of relapse patients underwent first ABMT with BAVC preparative regimen achieving a second CR without any remarkable complications. After 4, 5 and 4 months respectively a second ABMT was performed with a different preparative regimen, consisting of cyclophosphamide and fractionated total body irradiation (Cy + F‐TBI). A delay in platelet recovery was observed after the second procedure as compared to the first, while neutrophils recovery was comparable. 1 patient died in CR (on day +91 after second ABMT) of interstitial pneumonitis. 2 patients are alive and well without evidence of disease after 46 and 53 months of unmaintained second CR. This experience shows the high antieleukemic potential of treatment with a double sequential autograft; 2 relapsed patients in fact are long‐term survivors with a second CR longer than the first.
Bone marrow and blood involvement by non‐Hodgkin's lymphoma: A study of clinicopathologic correlations and prognostic significance in relationship to the Working Formulation
Authors: Morra Enrica , Lazzarino Mario , Castello Alessandro , Inverardi Daniela , Coci Anna , Pagnucco Guido , Orlandi Ester , Merante Serena , Magrini Umberto , Zei Gianna , Bernasconi Carlo
Published: July 2010

In a series of 172 patients with non‐Hodgkin's lymphoma (NHL) classified according to the Working Formulation (WF) the overall incidence of bone marrow infiltration (BM +) at diagnosis was 39%: 59% for low‐grade (LGML), 30% for intermediate‐grade (IGML), and 25% for high‐grade malignant lymphomas (HGML). The features most significantly correlated with the presence of BM + were a low grade of histological malignancy, the degree of splenomegaly and high values of LDH, while those correlated with the extent of BM+ were a non‐focal pattern of BM disease, the presence of blood involvement at diagnosis, and the degree of BM fibrosis. Blood involvement was detected at diagnosis in 13% of patients, and a further 16% developed a leukemic phase during the course of the disease. Blood involvement correlated significantly with splenomegaly, bulky disease, advanced clinical stage, and extent of BM +. The presence of BM infiltration ‘per se’ at diagnosis did not significantly affect prognosis. However, the extent of BM disease was correlated with a poorer outcome in IGML and HGML patients. Regarding peripheral blood involvement, in LGML patients only late leukemic conversions were significantly associated with a worse prognosis. In patients with IGML and HGML, either initial or subsequent blood involvement was correlated with significantly poorer outcome.
Ferritin synthesis in peripheral blood lymphocytes following in vitro stimulation*
Authors: Pattanapanyasat Kovit
Published: July 2010

The incubation of normal peripheral blood lymphocytes with phytohaemagglutinin results in an increase in DNA synthesis accompanied by an increase in ferritin synthesis, as determined by 3H‐leucine incorporation into ferritin isolated by immunoprecipitation. The increase was 5 to 8 times greater than that of non‐stimulated cells. This phenomenon was associated with a 3‐ to 6‐fold increase of intracellular ferritin when compared with the original ferritin concentration. While the significance of increased ferritin expression in proliferative cells is not known, an increase in ferritin synthesis following in vitro stimulation is probably related to increased metabolic activity of the cells.
Chronic myelomonocytic leukemia — clinicobiological characteristics: A multivariate analysis in a series of 70 cases
Authors: del Cañizo M. C. , Sanz G. , Miguel J. F. San , Vallespi T. , Irriguible D. , Torrabadella M. , Sanz M. A.
Published: July 2010

In a series of 70 patients diagnosed according to the FAB criteria, 42 clinical and biological disease characteristics were analyzed in order to identify significant prognostic factors by means of univariate and multivariate analysis. The univariate analysis identified ten parameters associated with poor prognosis: Symptoms of anemia, WBC over 10 × 109/1, presence of blast cells, myeloid precursors or erythroblasts in peripheral blood (PB), high bone marrow (BM) cellularity, severe dysthrombopoiesis, percent of blast cells in BM and high serum levels of bilirubin and LDH. The Cox proportional hazards regression method revealed that the combination of high leukocyte counts and BM percentage of blast cells had the strongest predictive relation to survival length (p = 0.002 and p = 0.060 respectively). A new multivariate analysis treating the presence of myeloid and erythroid precursors in PB as a single variable selected only this as the most significant prognostic factor (p = 0.001). Both regression models allowed us to discriminate two significantly different risk groups of patients.
Temperature‐dependent formation of vacuolar and canalicular systems in association with degranulation in stimulated neutrophils
Authors: Hara Yoji , Tsukada Masashige , Komiyama Atsushi , Akabane Taro
Published: July 2010

Effects of temperature on the morphological changes and degranulation were studied in stimulated neutrophils. Neutrophils were stimulated by formyl‐methionyl‐leucyl‐phenylalanine (FMLP) and cytochalasin B at 4°C, 20°C and 37°C, and they were processed for electron microscopy and biochemical assays of myeloperoxidase (MPO) activity. When neutrophils were stimulated at 4°C, they did not show apparent morphological changes nor release MPO activity. In the cells stimulated at 20°C, there were many cytoplasmic vacuoles, localization of MPO‐positive granules adjacent to the cell membrane and around the vacuoles, MPO‐positive materials in the cavity of the vacuoles, and slightly decreased numbers of granules. They released moderate amounts of MPO activity. The neutrophils at 37° C showed formation of small vacuoles and canaliculi and a marked decrease in the number of MPO‐positive granules indicative of degranulation, and they released increased amounts of MPO activity. There were MPO‐positive materials on the membrane and in the cavity of the vacuoles. Ruthenium red staining showed that the vacuoles were formed open outside the cell. These results indicate that the vacuolar systems are involved in degranulation in stimulated neutrophils and that their formation is dependent on temperature.
Peripheral blood stem cells collected before and after leukapheresis in the very early remission phase of hematopoietic malignancies
Authors: Vannier J. P. , Sumereau‐Dassin E. , Bastit D. , Jean P. , Tilly H. , Monconduit M. , Piguet H.
Published: July 2010

Human peripheral blood obtained after chemotherapy‐induced remission in hemopoietic malignancies has been suggested to be a potential substitute for autologous bone marrow as regards autologous hematopoietic reconstitution. The schedule and consequences of early leukapheresis are, however, still imprecise. We report a study performed in two series of, respectively, 10 and 14 patients where sequential leukapheresis (total number = 84) was evaluated with regard to colony‐forming unit (CFU) potency. Our data demonstrate that adequate numbers of progenitor cells can be collected by leukapheresis and that, even when this is performed at an early stage after remission, subsequent hematopoietic reconstitution is not impaired.
Radiolabelling of human platelets with 99mTc‐HMPAO
Authors: Vorne M. , Honkanen T. , Karppinen K. , Röning M. , Sakki S.
Published: July 2010

The optimum conditions for labelling platelets with lipophilic 99mTc‐hexamethyl propylene amine oxime (99mTc‐HMPAO) were evaluated. An aseptic closed system was used throughout the procedure in patient studies. 48 ml blood were withdrawn into 12 ml ACD using sterile 60 ml plastic syringes. After mixing, the blood was transferred to sterile 10 ml vacuum tubes and platelets were isolated according to standard centrifugation procedures. The labelling efficiency was not dependent upon incubation temperature (22°C, 37°C) but was greater in saline than in the presence of plasma. The labelling efficiency increased with time up to 60 min in saline. The elution of 99mTc from platelets was about 25% in plasma milieu in vitro but did not increase with time during 160 min. 5 patients with verified fresh deep vein thrombosis in the lower leg were imaged after injection of labelled autologous platelets. All 4 of the patients without anticoagulant therapy showed positive uptake of 99mTc‐HMPAO‐labelled platelets, but the 5th patient — under heparin therapy — was negative in scintigraphy. Our results are encouraging and 99mTc‐HMPAO‐labelled platelets offer a promising tool for evaluating various clinical situations.
Chronic myelomonocytic leukemia with chromosomal changes involving lp36 and hepatocellular carcinoma in a case of Fanconi's anemia
Authors: Bessho Fumio , Mizutani Shuki , Hayashi Yasuhide , Moriwaki Kohichi , Yokota Shunichiro , Inaba Toshiya
Published: July 2010

A girl with Fanconi's anemia developed chronic myelomonocytic leukemia and hepatocellular carcinoma. At the time that chronic myelomonocytic leukemia was diagnosed, all metaphases of the bone marrow cells revealed a chromosomal translocation involving 1p36. It is suggested that the occurrence of this rare type of leukemia in our patient is related to the chromosomal translocation.
Marrow aplasia developing 3 years after treatment with busulphan for chronic myeloid leukaemia
Authors: Bowcock S. J. , Galton D. A. G. , Goldman J. M.
Published: July 2010

A 31‐year‐old woman with Philadelphia chromosome‐positive chronic myeloid leukaemia (CML) was treated intermittently with high‐dose busulphan over a 6‐yr period (total dose 1320 mg). 3 yr later (after receiving no further cytotoxic drugs) she developed pancytopenia and marrow aplasia of relatively abrupt onset. Transfusion of reconstituted blood‐derived stem cells (collected 7 yr previously) re‐established chronic phase CML. These events are more consistent with ‘stem cell exhaustion’ than with an acquired marrow microenvironmental defect occurring in the course of CML. The contribution of busulphan is uncertain.
High‐dose methylprednisolone for remission induction in children with acute nonlymphoblastic leukemia
Authors: Hiçsönmez Gönül , Özsoylu Sinasi , Gürgey Aytemiz , Zamani Vahide P. , Irken Gülersu
Published: July 2010

The effectiveness of high‐dose intravenous methylprednisolone (HIVMP) in inducing an initial remission was examined in a child with acute nonlymphoblastic leukemia (ANLL). Dramatic clinical and hematological improvement with 7% marrow blasts was obtained in 3 weeks with HIVMP treatment without giving any other antileukemic drugs. Based on the results of this preliminary observation we suggest that high‐dose methylprednisolone (20–30 mg/kg) might be a useful approach when applied as an initial short treatment in childhood ANLL.
Phenotypic and functional characterization of normal and malignant terminal B (plasma) cells
Authors: Anderson Kenneth C. , Cochran Maria , Barut Bruce A.
Published: July 2010

A number of antigens (Ags) are expressed on normal and malignant terminal B (plasma) cells, including plasma‐cell, earlier B‐cell, and non‐B cell‐Ags. These Ags, coupled with indirect and dual fluorochrome labelling techniques, permit characterization of normal and malignant in vitro and in vivo terminal B‐cell differentiation. The majority (90%) of B cells within spleen bear B1 and lack PCA‐1 Ags. As B cells differentiate to pokeweed mitogen in vitro, immunoglobulin (Ig) secretion precedes the appearance of cell surface PCA‐1 and plasmacytoid morphology. Dual fluorescence cell sorting permits characterization of in vivo B‐cell differentiation: B1 + PCA‐1 + cells are more “differentiated” since they are more prevalent in lymph node than spleen, exhibit plasmacytoid morphology and maximal Ig secretion, and no longer respond to triggers of B‐cell proliferation; in contrast, B1 + PCA‐1‐ cells are lymphoid in morphology and may respond to triggers of B‐cell proliferation as “resting” B cells. Similar studies of myeloma cells demonstrated that they may also include cells expressing plasma‐cell, earlier B, and non‐B cell Ags. Although they neither proliferated nor secreted Ig in vitro to G/M‐CSF, G‐CSF, M‐CSF, IL‐1, IL‐1B, IL‐2, or IL‐4, proliferation without Ig secretion (Stimulation Index ≥ 3.0) was induced to IL‐6 in 6 of 10 patients (pts); to IL‐3 (2 pts) and to IL‐5 (2 pts). Myeloma cells which proliferated to the greatest extent to IL‐6 have plasmablast morphology and appear to express an earlier B‐cell phenotype (B4±CALLA±B1 + PCA‐1 ±) than those cells which either did not or only weakly responded (B4‐CALLA ‐B1±PCA ‐1±). Phenotypic definition of growth factor‐responsive myeloma cells may both facilitate their biological characterization in vitro and permit enhanced understanding of aberrant B‐cell differentiation in vivo.
Circulating malignant precursors in monoclonal gammopathies
Authors: Caligaris‐Cappio F. , Bergui L. , Gaidano G. L. , Schena M. , Putto P. , Merico F. , Riva M.
Published: July 2010

Peripheral blood mononuclear cells (PBMC) from 14 patients with multiple myeloma (MM), 3 patients with benign monoclonal gammopathy, 3 patients with Waldenstrom's macroglobulinaemia (WM) and 2 patients with B‐cell chronic lymphocytic leukemia (B‐CLL) were cultured in vitro in the presence of IL‐3 and IL‐6. After 3 days, actively proliferating immunoblast‐like B cells were apparent in 12/14 cases of MM, 0/3 BMG, 3/3 WM and 0/2 B‐CLL. After 6 d, B blasts had evolved into morphologically evident plasma cells expressing the specific monoclonal light and heavy chains. The data indicate that the concerted action of IL‐3 and IL‐6 synergistically promotes the proliferation and differentitation of circulating plasmacell precursors in malignant monoclonal gammopathies.
Advances in biology of multiple myeloma: Cell kinetics, molecular biology and immunology
Authors: Pileri Alessandro , Ferrero Dario , Massaia Massimo , Dianzani Umberto , Boccadoro Mario
Published: July 2010

Bone marrow plasma cell proliferative activity has been evaluated in a large series of multiple myeloma (MM) patients. This kinetic parameter has been shown to be a useful tool for patient management, and contributes to a correct diagnosis and a selection of high‐risk patients who can be offered high‐dose chemotherapy. The role of ras oncogenes has been evaluated in the pathogenesis of MM. A point‐mutated and activated H‐ras oncogene, introduced in a human lymphoblastoid cell line, was able to induce neoplastic transformation and differentiation to plasma cell. Indeed, mutated alleles of ras genes have been detected in a high percentage of myeloma patients in relapse phase. Phenotypical and functional studies have been carried out in T‐lymphocyte subsets and an impaired cellular immunity has been detected. Such an impairment was related to the disease status: marked alterations were detected in relapse phase, whereas a partial recovery was observed during remission phase.
Chromosomes in plasma‐cell malignancies*
Authors: Van Den Berghe Herman
Published: July 2010

Mush less is known about the chromosome changes in MM than in other hematological malignancies. The prevalence of abnormal karyotypes is unknown, but there is no evidence for malignant plasma cells or their precursors to have a normal karyotype. The chromosome changes found may be early events, but karyotypic evolution occurs early and rapidly. No specific structural or numerical chromosome anomaly is associated with multiple myeloma or plasma‐cell leukemia. The changes found are those already known to occur in other B‐cell malignancies, particularly B‐CLL and diffuse small cell lymphoma. A 14Q + marker is present in about 30% of all karyotypically abnormal cases, and in 50% of the cases this is due to a t(11;14) (q13;q32). In a minority of cases deletions of 6q are found, and sporadically other B‐cell translocations can be present. Karyotypes are often very complex with numerous structural anomalies involving mainly chromosomes 1, 11 and 17, and numerical anomalies involving chromosomes 3, 7, 9 and 11. Finally, the presence of structural or numerical anomalies of chromosomes 5 and 7 may be heralding or may be indicative of therapy‐induced leukemia.
Histology and immunohistology of bone marrow biopsy in multiple myeloma
Authors: Pileri S. , Poggi S. , Baglioni P. , Montanari M. , Sabattini E. , Galieni P. , Tazzari P. L. , Gobbi M. , Cavo M. , Falini B. , Stein H. , Tura S.
Published: July 2010

B5‐fixed/paraffin‐erabedded Jamshidi needle biopsies from 125 multiple myeloma patients were reviewed according to both morphological and immunohistological criteria. At microscopic examination, the following parameters were evaluated: i) grade of malignancy (low = 56; intermediate = 50; high = 19); ii) growth pattern (interstitial + /‐ sheets/nodules = 90; nodular =13; packed marrow = 18; sarcomatous = 4); III) histological stage (I = 64; II = 35; III = 26). Comparison of the findings in trephine biopsies and aspirates showed that in 30% of the cases the latter led to an understimation of the tumor burden. Immunohistochemical determination of Ig easily allowed: i) differential diagnosis from exuberant reactive plasmacytosis; ii) recognition and counting of neoplastic plasma cells; iii) detection of minimal residual disease after treatment. Immunohistochemistry also confirmed phenotypic aberration of neoplastic plasma cells, showing positivity for CD45, EMA, and cytokeratins in 14%, 59%, and 25% of the cases, respectively. Furthermore, it displayed expression of the P‐glycoprotein in 4/8 resistant cases. These findings underline that routinely processed Jamshidi needle biopsies can be of great value in the study of patients with multiple myeloma.
Renal failure in myelomatosis
Authors: MacLennan I. C. M. , Cooper E. H. , Chapman C. E. , Kelly K. A. , Crockson R. A.
Published: July 2010

Renal failure is a common presenting feature in myelomatosis. This review offers a practical means for classifying renal failure in this disease. Three groups are identified: (1) those patients whose renal failure improves or is stable when they are maintained on a high fluid intake; (2) the minority of patients whose renal failure progresses despite high fluid intake; and (3) those patients who are fluid‐intolerant due to oligurlic renal failure or congestive cardiac failure. The difference beween groups 1 and 2 is not simply due to differences in response to chemotherapy, for many group‐1 patients achieve improvement in renal function without or before loss of light chain proteinuria. It is concluded that all patients with myelomatosis with excess monoclonal free light chain proteinuria are at risk from developing renal failure of the type associated with group 1. The chances of them doing so are diminished if they maintain a high fluid intake. Group 2 encompasses a range of conditions not all of which are clearly defined. There is generally a poor correlation between the physical characteristics of light chains and the presence of group‐2 renal failure.
Monoclonal gammopathy of undetermined significance and smoldering multiple myeloma
Authors: Kyle Robert A.
Published: July 2010

Patients with monoclonal gammopathy of undetermined significance (MGUS) have a monoclonal protein but no evidence of multiple myeloma, macroglobulinemia, amyloidosis, or related disorders. In a long‐term follow‐up (median, 19 years) of 241 patients with MGUS, 53 (22%) developed multiple myeloma (36 patients), macroglobulinemia (7 patients), primary systemic amyloidosis (7 patients), or malignant lymphoproliferative disease (3 patients). Fifty‐seven patients (24%) remained stable and were alive, while 124 patients (51%) died of causes unrelated to the monoclonal gammopathy. Multiple myeloma was diagnosed in the 36 patients 23 to 251 months (median, 9.6 years) after recognition of the M‐protein. Smoldering multiple myeloma (SMM) is characterized by a serum M‐protein level > 30 g/1 and > 10% plasma cells in the bone marrow but no anemia, renal insufficiency, or lytic lesions. Differentiation of MGUS and SMM from multiple myeloma and macroglobulinemia is difficult at the time of diagnosis of the M‐protein, and the patients must be followed up indefinitely.
Plasma cell leukemia: A report on 15 patients
Authors: Bernasconi C. , Castelli G. , Pagnucco G. , Brusamolino E.
Published: July 2010

Plasma cell leukemia (PCL) can be considered the leukemic variant of multiple myeloma. The diagnosis is based on hematological features, including a plasmacytosis exceeding 2 × 109/l and any evidence of a clonal plasma cell proliferation. There are two forms of PCL: the primary form occurring in individuals without preceding multiple myeloma, and the secondary form arising as a late manifestation in patients with multiple myeloma. From 1974 to 1988 we diagnosed 8 primary PCL cases out of a total 301 multiple myeloma cases (incidence, 2.6%) and a total of 847 acute leukemia cases (incidence, 0.9%). During the same period we observed in 7 multiple myeloma patients a terminal PCL, for an incidence of PCL in myeloma of 2.3%. Most clinical characteristics were similar in both types of plasma cell leukemia. In particular we found no difference in the average age and in the incidence of bone pain, hepatosple‐nomegaly, lytic bone lesions. None of our cases showed a clinically relevant lymphade‐nopathy either as presenting symptom or during the course of the disease. The values for hemoglobin, leukocytes, plasma cells, serum creatinine and calcium did not differ significantly between the two groups of patients. The median survival was 7 months for patients with primary PCL and 1 month for patients with secondary PCL. 5 of the 8 patients with primary PCL obtained a response to conventional myeloma therapy including single alkylating agents, with a duration ranging from 7 to 44 months. Only 1 of the patients with secondary PCL had a partial response after combination chemotherapy.
Bone marrow histology and serum beta 2 microglobulin in multiple myeloma — a new prognostic strategy
Authors: Bartl R. , Frisch B. , Diem H. , Mündel M. , Fateh‐Moghadam A.
Published: July 2010

Bone marrow biopsies of 720 patients with multiple myeloma (MM) were investigated from 1968 — 1989. Histologic variables were correlated with clinical parameters and survivals to determine prognostic factors. In 207 of these patients initial levels of serum beta 2 microglobulin (SB2M) were also measured for prognostic evaluation. Four tumour growth patterns were distinguished: interstitial (56%), interstitial/sheets (13%), nodular (15%) and packed marrow (16%) with median survivals of 46, 31, 22 and 16 months. When grouped according to the tumour cell mass in the biopsy, four histologic stages were recognized. Cellular characteristics were used to classify MM into 6 histologic types which were subsequently combined into 3 grades of malignancy: low, intermediate and high, analogous to the malignant lymphomas. With respect to tumour products, only SB2M proved to be a valuable prognostic indicator for staging and follow‐up. Complications of MM such as anaemia, azotaemia, osteolytic lesions, hypercalcaemia and hypoalbuminaemia all predicted a poor prognosis, highly significant in the test statistics. We propose a new prognostic approach in MM, comprising 1) parameters defining the tumour itself, 2) the tumour products (SB2M) and 3) the tumour complications. This prognostic strategy combines information both on stage and malignancy of MM and enables definition of smouldering and of aggressive variants of MM at an early stage.
Improvement of Durie & Salmon staging for multiple myeloma by adding platelet count as a stratifying variable: A multivariate regression analysis of 163 untreated patients
Authors: Cavo Michele , Galieni Piero , Grimaldi Maria , Zuffa Eliana , Bonelli Maria Angela , Gobbi Marco , Tura Sante
Published: July 2010

The presenting clinical features of 163 previously untreated patients with multiple myeloma were correlated with survival duration using univariate and multivariate regression analyses. The univariate proportional hazard analysis ranked the parameters in the following order of importance: platelet count, haemoglobin level (Hb), tumour cell mass stage, lytic bone lesions, creatinine and age. When the individual contribution of each variable was assessed by multivarite regression analysis, platelet count was confirmed to be the dominant feature for prognosis, while clinical stage provided additional information. The introduction of platelet count could then be used to improve the discriminating power of Durie & Salmon staging, by allowing separation of the high‐risk group (stages II and III) into a smaller subgroup (22%) of thrombocytopenic patients (< 150 × 109platelets/l) whose risk of death was actually very high (median survival: 9 months) and a larger subgroup (46%) of patients with normal platelet count and intermediate or standard risk (median survival: 48 months).
The Merlini, Waldenström, Jayakar staging system revisited
Authors: Merlini Giampaolo , Gobbi Paolo G. , Ascari E.
Published: July 2010

At present, none of the routinely used staging systems is entirely satisfactory, since all of them require some reconsideration to account for the most recent prognostic factors. Among these, serum β‐2 microglobulin (S‐β‐2M) has proved to be a powerful prognostic test assessable in most centers. In order to improve the Merlini, Waldenström, Jayakar (MWJ) staging system, we have determined the prognostic significance of several factors in an independent population of 345 retrospective myeloma patients seen at our institution from 1973 to 1988. The Cox regression hazards method showed that S‐Creatinine, S‐Albumin levels and percentage of bone marrow plasma cells accounted for 94% of total χ2achieved with all eight variables (including also: paraprotein index, S‐Calcium, S‐M‐component, bone lesions and hemoglobin). The same analysis, applied to patients in whom S‐β‐2M was available at presentation, selected S‐β‐2M, S‐Creatinine and percentage of bone marrow plasma cells (accounting for 92% of total χ2), followed by platelet count, S‐Albumin and S‐M‐component. The best discriminatory level of S‐β‐2M was found to be 4 mg/***l. However, the prognostic power of S‐β‐2M when used with binary values was remarkably reduced with respect to its application as a continuous variable. In patients with MWJ stage 1 and 2 disease, S‐β‐2M had an independent prognostic significance. Grouping stage I and II patients with S‐β‐2M less than or more than 4 mg/1 presented a median survival of 70 and 40 months, respectively. In stage 3 patients S‐β‐2M did not contribute additional prognostic information. The integration of S‐β‐2M, as a continuous variable, into the MWJ staging system may allow a better assessement of individual death risks.
Therapeutic implications of myeloma staging*
Authors: Durie Brian G. M. , Bataille Regis
Published: July 2010

Clinical staging, such as with the Durie‐Salmon clinical staging method, remains the most easily usable available method to classify patients with untreated multiple myeloma. However, for the development of new treatment strategies and comparison with prior results, new and powerful prognostic factors are now available. Serum beta 2 microglobulin alone represents the most reliable prognostic factor in multiple myeloma. Beta 2 microglobulin alone allows simple and reproducible classification of patients into low grade myeloma with low serum beta 2 microglobulin versus high grade myeloma with high serum beta 2 microglobulin. Plasma cell labelling index is an additional factor which allows identification of patients with MGUS as well as subclassification of patients with low serum beta 2 microglobulin into those with relatively better or worse prognosis. For each grade of myeloma young (less than 63 years of age) patients have a slightly better survival. Independently of serum beta 2 microglobulin and labelling index, DNA content values allow the identification of a subset of patients with biclonal or hypodiploid tumors and high grade disease. New or more sophisticated methods for evaluating myeloma cells such as by immunopheno‐typing or assessment of multi drug resistance enable the development of specific approaches to treatment in individual cases. It should also prove possible to identify patients especially suitable for newer biologic agents such as alfa interferon, G or G‐MCSF and the various interleukins.