bluuebunny/crossref_metadata_2025 huggingface dataset:
Confirmation of bone marrow engraftment by detection of M/N blood group antigens on erythroblasts in erythroid bursts
Authors: Kitano Kiyoshi , Kajii Eiji , Ikemoto Shigenori , Amemiya Yoichi , Komatsu Norio , Suda Toshio , Miura Yasusada
Published: March 2008

In order to confirm erythroid engraftment, we examined the changes in blood group antigens expressed on erythroblasts in the haematopoietic colonies of a bone marrow transplant recipient. A 44‐year‐old female with acute myelogenous leukaemia underwent an allogeneic bone marrow transplantation from her sister donor. Prior to the transplantation, the blood groups of both recipient and donor were analysed, and their M/N groups found to differ, the former being MN and the latter N. Bone marrow mononuclear cells were obtained from the patient on day 21 after bone marrow transplantation and cultured in semi‐solid medium. Erythroblasts were collected from the erythroid bursts that had formed, and were subjected to flow cytometry using monoclonal anti‐M and anti‐N. Anti‐N reactive cells accounted for 99.2% of those in the erythroid bursts, while only 0.3% of these cells were anti‐M reactive. MN type erythrocytes in the recipient's peripheral blood had been replaced by N type and only 2.0% of erythrocytes were anti‐M reactive on the 70th day after BMT. These erythroblasts and erythrocytes were phenotypically N, and originated from the donor haemopoietic stem cells with the success of bone marrow engraftment. From our findings, it appears that the M/N blood group antigens were produced by the erythroid cells themselves.
Selective growth response to IL‐3 of a human leukaemic cell line with megakaryoblastic features
Authors: Avanzi Gian Carlo , Lista Patrizia , Giovinazzo Bruna , Miniero Roberto , Saglio Giuseppe , Benetton Gabriella , Coda Renato , Cattoretti Giorgio , Pegoraro Luigi
Published: March 2008

A new human leukaemic cell line (M‐O7) with the phenotypic characteristics of CFU‐mega is described. Its cells are positive for T200 leucocyte common antigen (LCA) and negative with MAbs recognizing T and B cells and mature myelomonocytic antigens. In contrast, they react with MAbs recognizing antigenic determinants common to multilineage (CD13, CD33, CD34) and to bipotent erythromegakaryoblastic (CD36, H25) haemopoietic precursors, and with MAbs specific for platelet glycoproteins (CD41w, CD42w). A small proportion (10%) of the cells were large and multinucleated, and on electron microscopy examination showed peripheral splitting of platelet‐like cytoplasm particles. When transferred to a serum‐free Iscove modified Dulbecco's medium supplemented with human insulin and transferrin, M‐O7 cells stop proliferating. Of the haemopoietic growth factors tested for their ability to restore the proliferative activity of this quiescent population, only rH IL‐3 proved effective. Moreover, it also increased the cloning efficiency in methylcellulose more than any other CSFs. The M‐O7 cell line may provide a valuable tool for the biological assay of IL‐3, and a model for biochemical studies of the megakaryocytic lineage.
Testosterone reduces complement sensitivity of precursor cells in aplastic anaemia patients treated with antilymphocyte globulin
Authors: Nissen Catherine , De Planque Mariet M. , Würsch A. , Moser Yolanda , Carbonare Verena Dalle , Speck B.
Published: March 2008

Of 53 consecutive patients with aplastic anaemia who were re‐examined at various intervals after treatment with antilymphocyte globulin, 30 had sufficient bone marrow colony forming capacity to permit evaluation of androgen effects in vitro. In 22 patients, precursor cells of the myeloid and erythroid line were abnormally sensitive to a preincubation in isosmolar sucrose with 5% fresh autologous serum compared to heat‐inactivated autologous serum. This phenomenon was interpreted as excess complement sensitivity. This inhibitory effect of fresh serum in the bone marrow sucrose test was abrogated by addition of 10‐6 M testosterone to the preincubation phase in 15 of the 22 patients. In six of these 15, 10‐7 M dexamethasone had a similar effect: in the other nine patients only testosterone rendered the bone marrow sucrose test negative. This effect of testosterone on colony growth was indirect, since addition of 10‐9‐10‐5 M testosterone to primary bone marrow cultures from the same patients had no effect. We propose that testosterone in these experiments interacted with the complement system. In patients who have complement sensitive precursor cells, androgens might thus prevent complement mediated lysis of haemopoietic cells to some extent. The test described could help identification of patients in autologous bone marrow remission who are likely to benefit from androgen treatment.
Human megakaryocyte size varies with anatomical site
Authors: Gladwin A. M. , Martin J. F. , Thorpe J. A. C. , Trowbridge E. A.
Published: March 2008

Whole megakaryocyte, megakaryocyte cytoplasmic and nuclear sizes were evaluated in human bone marrow obtained from posterior iliac crest and rib of subjects undergoing thoracotomy for repair of hiatal hernia. In addition the nuclear DNA content (ploidy) of the cells from each site was measured. Significantly larger mean megakaryocyte (P < 0.002), and megakaryocyte cytoplasmic volumes (P < 0.002), and different volume distributions were observed in costal marrow compared to the iliac crest. No change in mean nuclear volume was found. This was paralleled by no significant difference in mean ploidy. This suggests that the variation in megakaryocyte size with anatomical site is determined by the concomitant volume of cell cytoplasm, but not the nuclear volume.
Prospective confirmation of a prognostic classification for Ph + chronic myeloid leukaemia
Authors: Russo Domenico
Published: March 2008

In 1984, Sokal et al described a prognostic model for Ph + chronic myeloid leukaemia (CML), based on retrospective analysis of data from six series of patients in the United States and Europe. In the same year, a prospective study of prognosis was started by the Italian Cooperative Study Group on CML. The first analysis of the prospective study is based on 508 Ph +, non‐blastic, non‐transplanted patients registered between January 1984 and December 1986. Application of the hazard ratio formula described in 1984 to this prospective series has shown that the model predicts accurately the frequency and the time‐distribution of blastic transformation and death during the first 2 years after diagnosis. These results confirm the earlier conclusion that prognostic discrimination is possible at the time of diagnosis of CML, and indicate that the model described in 1984 can prove useful in evaluation and management of this disease.
Inheritance and bleeding in factor XI deficiency
Authors: Bolton‐Maggs P. H. B. , Wan‐Yin B. Young , McCraw A. H. , Slack J. , Kernoff P. B. A.
Published: March 2008

A study of 20 Jewish and four non‐Jewish kindreds transmitting factor XI deficiency (164 individuals) confirmed inheritance to be autosomal with severe deficiency in homozygotes (mean factor XI level 3.8 u/dl, SD 2.91) and partial deficiency in heterozygotes (mean factor XI level 57 u/dl, SD 10.42; normal mean factor XI level 96 u/dl, SD 11.6). The probability of an individual being heterozygous can be predicted from the factor XI level using a graph derived from this data. The accuracy is increased by including the prior probability derived from the pedigree. A high frequency of heterozygote to heterozygote mating was observed in the Jewish families consistent with an estimated gene frequency of 13.4% in this racial group. The relationship between factor XI level and bleeding tendency is poor; a third of heterozygotes had bled excessively after surgery, including six with factor XI levels above 50 u/dl, showing this condition to have clear signs of expression in heterozygotes. The lower limit of the normal range (2 SDs from the mean) was found to be 72 u/dl.
Analysis of the membrane glycoproteins of platelets in the Wiskott‐Aldrich syndrome
Authors: Pidard Dominique , Didry Dominique , Deist Françoise Le , Durandy Anne , Griscelli Claude , Bellucci Sylvia , Nurden Alan T.
Published: March 2008

We have examined the plasma membrane glycoproteins of platelets from three unrelated patients with the Wiskott‐Aldrich syndrome. Single‐ or two‐dimensional SDS‐polyacrylamide gel electrophoresis was performed. Glycoproteins were located by staining for carbohydrate, or by autoradiography when the platelets had been surface‐labelled with 125I prior to solubilization. In one patient a slight decrease in the 125I‐labelling intensity of GP Ib, GP la and a 125I‐labelled polypeptide of M, 168 000 were noted. For the two other patients the glycoprotein profiles were indistinguishable from those of normal subjects. These results clearly indicate that abnormalities in platelet membrane glycoproteins are not a common trait among Wiskott‐Aldrich patients, and thus cannot be regarded as primary defects in this disease.
Auto‐anti‐Jka in Evans’syndrome with negative direct antiglobulin test
Authors: Ganly P. S. , Laffan M. A. , Owen I. , Hows J. M.
Published: March 2008

A patient presented with haemolytic anaemia and a negative direct antiglobulin test (DAT), and was found to have an IgG antibody with anti‐Jka specificity in his serum. His red cells were typed as Jk(a‐b+). Later he developed idiopathic thrombocytopenic purpura (ITP), and had a positive DAT due to anti‐Jka bound to his red cells, which now typed as Jk(a+b+). Family studies suggested that the patient's true type was Jk(a+b+). Splenectomy and immunosuppression were required to treat the thrombocytopenia. The autoanti‐Jka was no longer detectable following therapy.
Essential thrombocythaemia and the Philadelphia chromosome
Authors: Morris C. M. , Fitzgerald P. H. , Hollings P. E. , Archer S. A. , Rosman I. , Beard M. E. J. , Heaton D. C. , Newhook C. J.
Published: March 2008

Six adult patients presented with clinical features of essential thrombocythaemia. Five of the patients, although Ph‐positive, have maintained these features without evidence of leukaemia; in one case for 9 years. A sixth patient developed leukaemic blast crisis following a persistently high platelet count over 4 years. Her cells were Ph‐negative, but hybridization of gene probes to chromosomes in situ and to leukaemic DNA showed that the abl oncogene had moved to the breakpoint cluster region (bcr) on the normal chromosome 22. This patient has the same molecular gene change as occurs in some cases of Ph‐negative chronic myeloid leukaemia (CML) whose leukaemic cells likewise show no evidence of chromosomal translocation. Molecular studies are essential for the correct diagnosis of these patients. The Ph genomic lesion appears to have a range of leukaemic expression which includes thrombocythaemia as well as chronic myeloid leukaemia and acute lymphatic leukaemia.
Factor VIII polypeptide specificity of monoclonal anti‐factor VIII antibodies
Authors: Shima M. , Yoshioka A. , Nakai H. , Tanaka I. , Fujiwara T. , Terada S. , Imai S. , Fukui H.
Published: March 2008

Four monoclonal antibodies against factor VIII, NMC‐VIII/1, NMC‐VIII/2, NMC‐VIII/3 and NMC‐VIII/4, were produced. The first three antibodies were of the IgG1 immunoglobulin subclass, while the fourth was IgM. The affinity of each antibody for factor VIII was high and antifactor VIII clotting activity was detected in NMC‐VIII/2, NMC‐VIII/3 and NMC‐VIII/4. NMC‐VIII/1 had no inhibitory effect on factor VIII clotting activity. Immunoblotting using purified intact and thrombin‐treated factor VIII identified the antibodies’factor VIII polypeptide specificities. With thrombin‐treated factor VIII the factor VIII fragments for NMC‐VIII/1 and NMC‐VIII/2 were 80 kDa and 54 kDa, respectively, while both NMC‐VIII/3 and NMC‐VIII/4 recognised a 44 kDa fragment. With intact factor VIII, antibodies NMC‐VIII/2‐4 bound to polypeptides larger than 90 kDa, especially NMC‐VIII/2, which reacted with a chain of 330 kDa thought to be a mature form of factor VIII protein. We also detected a mature form of factor VIII in hepatic sinusoidal endothelial cells by immunoperoxidase staining.

Identification of the factor VIII polypeptides bearing these fragments enabled us to clarify the localization of the factor VIII thrombin cleavage site, and suggested the existence of factor VIII in hepatic sinusoidal endothelial cells.
Relationship between the in vitro sensitivity to cytosine arabinoside of blast progenitors and the outcome of treatment in acute myeloblastic leukaemia patients
Authors: Nara N. , Suzuki T. , Nagata K. , Yamashita Y. , Murohashi I. , Adachi Y.
Published: March 2008

The sensitivity to cytosine arabinoside (Ara‐C) of blast progenitors from 22 acute myeloblastic leukaemia (AML) patients was studied in methylcellulose and suspension cultures. Primary colony‐formation (PE1) in methylcellulose reflects the terminal divisions of blast progenitors, while secondary colony formation (PE2) in methylcellulose and the clonogenic cells recovery in suspension are considered to be based on the self‐renewal of blast progenitors, In any patient, PE2 or clonogenic cells in suspension were more sensitive to Ara‐C than PE1. The results indicate that Ara‐C effectively suppresses not only terminal divisions but also selfrenewal of blast progenitors D10 Ara‐C value, the dose required to reduce survival to 10% of control, for PE1, PE2 and clonogenic cells in suspension showed marked patient‐to‐patient variation. No significant correlation was found between D10 Ara‐C in methylcellulose or suspension culture and the response to treatment with a combination chemotherapy of 6‐mercaptopurine, Ara‐C and daunorubicin. However, a relapsed patient whose D10 values in methyl‐cellulose and suspension cultures were very high showed poor response to a high‐dose Ara‐C protocol, where Ara‐C was given alone at a high dose. The application of chemo‐sensitivity test as a prediction of the clinical outcome may be dependent on the treatment protocol.
Defective megakaryocytopoiesis in the syndrome of thrombocytopenia with absent radii
Authors: Homans Alan C. , Cohen Janet L. , Mazur Eric M.
Published: March 2008

The syndrome of thrombocytopenia with absent radii (TAR) is a hereditary condition whose pathogenesis is poorly understood. In this investigation we evaluated a female infant with TAR and her parents using in vitro haematopoietic colony forming assays and an antiserum against platelet membrane glycoproteins (PGP) to label smears of her bone marrow. Megakaryocyte colony growth in vitro was virtually absent in optimally stimulated cultures of the patient's bone marrow progenitors. In contrast, erythroid and myeloid colony growth from the TAR infant's marrow cells was preserved. Staining of the patient's bone marrow smears with PGP antiserum detected no immature, small megakaryocyte precursors. A high level of megakaryocyte colony stimulating activity was detected in serum from the TAR infant, activity comparable to that present in sera from adults with aplastic anaemia. The elevated serum activity decreased by 6 months of age at which time partial platelet recovery had occurred. Evaluation of both peripheral blood haematopoietic progenitor cells and sera from the TAR infant's parents demonstrated no significant abnormalities. We conclude that the principle haematopoietic defect in this patient with TAR syndrome is the absence or arrested development of the committed megakaryocyte progenitor cell. Humoral regulation of megakaryocytopoiesis appears intact and is responsive to the degree of megakaryocytic hypoplasia.
Phenotypic and functional T cell subset abnormalities in patients with aplastic anaemia and hypogammaglobulinaemia
Authors: Raziuddin Syed , Sheikha Anwar K. I.
Published: March 2008

We have investigated T cell abnormalities present in blood of two patients with aplastic anaemia and hypogammaglobulinaemia. There was a marked increase in class II. major histocompatibility complex, HLA‐DR+ antigen, and interleukin‐2 receptor (Tac+) bearing CD4+ helper/inducer T cells, and a concurrent reduction of CD8+ suppressor/cytotoxic T cells. These CD4+ T cells produced an elevated proliferative response to phytohaemagglutinin and concanavalin A. Interestingly, the T cell subset mainly responsible for elevated production of the lymphokine, interkeukin‐2, under the stimulus of phytohaemagglutinin, was characterized as belonging to a CD4+ T cell subset. Functional studies, using a pokeweed mitogen driven IgG, IgA and IgM synthesis, demonstrated a correlation between CD4+ T cell deficient helper function for B cell differentiation and the clinical finding of the patient's hypogammaglobulinaemia.
Treatment of idiopathic thrombocytopenic purpura with ascorbate
Authors: Brox Alan G. , Howson‐Jan Kang , Fauser Axel A.
Published: March 2008

The treatment of idiopathic thrombocytopenic purpura (ITP) includes corticosteroids, danazol, splenectomy and various immunosuppressives. Treatment can be difficult for those patients refractory to these modalities and/or those patients intolerant of the secondary effects. In this paper we report on the use of ascorbate in the treatment of ITP and its successful use in seven of 11 patients studied. We found that therapy with ascorbate appeared to improve the platelet count and the intravascular survival of platelets. Because of excellent patient compliance and its lack of toxicity, it may be an alternative for the treatment of ITP. The exact role of ascorbate in the treatment of ITP, as well as its mechanism of action, await further study.
Thrombovascular disease and familial plasminogen deficiency: a report of three kindreds
Authors: Dolan G. , Greaves M. , Cooper P. , Preston F. E.
Published: March 2008

We report three kindreds in whom plasminogen deficiency was associated with thrombosis and in whom the ratio of functional and immunological values of plasminogen was consistent with type I deficiency. An additional subject with plasminogen deficiency is also described. The three propositi presented with venous thrombotic disease. The fourth subject presented with a thrombotic stroke. Investigation of family members in three of these four cases revealed other subjects who were found to have low levels of plasminogen and who were asymptomatic. The pattern of inheritance appears to be autosomal dominant. In one woman, plasminogen levels were shown to rise to within the normal range during pregnancy and returned to low levels after delivery. A total of eight pregnancies were reviewed in our series and no thrombotic events occurred.
Production of mouse monoclonal antibodies for the analysis of idiotypes in serum of patients with chronic lymphatic leukaemia
Authors: De Rie Menno A. , Van Heemstra Diederik J. M. , Huijgens Peter C. , Zeijlemaker Wim P. , Out Theo A. , Melief Cornelis J. M. , Von Dem Borne Albert E. G. Kr.
Published: March 2008

Summary. In this report a simple procedure for the production of murine monoclonal antibodies (MoAb) against the idiotype of malignant B cells is described. Mice were immunized with lymphoid cells from patients with B‐cell chronic lymphocytic leukaemia (B‐CLL). After fusion of the spleen cells, hybridoma supernatants were screened for anti‐idiotypic MoAb in ELISA with immunoglobulins obtained from tumour‐cell lysates, xenohybridomas and patients' sera. The anti‐idiotypic MoAb were used to study tumour cells and serum immunoglobulins (Ig) from four different patients with B‐CLL. It was found that the serum IgM and IgD in one patient shared the same idiotype. Evidence is presented that IgG‐secreting cell populations are not restricted to λ‐Ig‐light chain‐expressing B‐CLL cells. With the help of anti‐idiotype MoAb accurate measurements of total and idiotype‐positive serum immunoglobulin levels during chemotherapy were possible.
Induction of myeloperoxidase in five cases of acute unclassified leukaemia
Authors: Heil G. , Ganser A. , Raghavachar A. , Kurrle E. , Heit W. , Hoelzer D. , Heimpel H.
Published: March 2008

Summary. For further analysis of their lineage affinity the differentiation capacity of five cases of acute unclassified leukaemias were studied in vitro using 5 nm 12‐O‐tetradecanoyl‐phorbol‐13‐acetate as an inducing agent. After 4 d in vitro a co‐expression of both myeloid and early B‐lymphoid or myeloid and early T‐lymphoid antigens on the same population of cells was found. While ultrastructural analysis of the blasts prior to culture revealed the morphology of undifferentiated blasts without any specific endogenous peroxidase activity, the cultured blasts displayed monoblastic features and myeloperoxidase activity after 4 d in vitro. Since this type of peroxidase is highly specific for the myelomonocytic lineage the data indicate myelomonocytoid differentiation of all five cases studied. The co‐expression of lymphoid as well as myeloid antigens and myeloperoxidase by the same blasts after culture could be indicative of bilineage differentiation capacity characterizing the original blasts as bipotential progenitor cells.
The development of hypoprothrombinaemia following antibiotic therapy in malnourished patients with low serum vitamin K1 levels
Authors: Cohen H. , Scott S. D. , Mackie I. J. , Shearer M. , Bax R. , Karran S. J. , Machin S. J.
Published: March 2008

Summary. A group of nine well‐nourished patients, with normal serum vitamin K1 levels (mean 546, range 310–1350 pg/ml), maintained normal prothrombin times (PTs) and factor VII clotting activities throughout a 7 d course of i.v. cefotetan disodium, an N‐methyl‐thiotetrazole (NMTT) containing cephalosporin antibiotic. However, 11 of 20 patients, with acute intra‐abdominal sepsis and initially normal PTs who underwent emergency surgery, developed prolonged PTs (INR 1.4–3.1) associated with reduction in factor VII activities (0.74–0.38 u/ml) after 3–7 d of antibiotic therapy. Nine of these 11 patients had clinical evidence of malnutrition and nine had subnormal serum vitamin K1 levels (mean 119, range 43–354 pg/ml) on admission. Seven received cefotetan but four were treated with a non‐NMTT‐containing cephalosporin or antibiotics belonging to other groups. The nine patients who maintained normal PTs all had normal nutritional status and normal serum vitamin K1 levels (mean 279, range 103–915 pg/ml) at presentation. The PT is a relatively insensitive indicator of vitamin K stores, and malnourished patients with low serum vitamin K1 levels are at risk of developing hypoprothrombinaemia following intravenous antibiotic therapy.
Selective decrease in platelet dense granule adenine nucleotides during recovery from acute experimental thrombocytopenia and ensuing thrombocytosis in baboons
Authors: Savage Brian , Hanson Stephen R. , Harker Laurence A.
Published: March 2008

Summary Serial measurements of platelet volume, platelet content of adenine nucleotides, β‐thromboglobulin (β‐TG), platelet factor 4 (PF4) and ex vivo platelet aggregation were made in baboons under basal, steady‐state conditions of normal platelet production, and during recovery from acute thrombocytopenia induced by the exposure of flowing blood to spherical glass microbeads. The mean basal platelet count of 509.107X109/l (±1 SD; n=4) fell acutely to 36.8±12.6X109/l after the insertion of glass bead columns and blood filters, placed distally, for 60 min into surgically implanted arteriovenous‐shunts in heparinized baboons. After the irreversible removal of up to 90% of the baseline circulating platelet population, recovery from thrombocytopenia was characterized by a constant rate of increase in circulating platelet counts (115±11X109/l/d) and a rebound thrombocytosis to 1.5 times the basal platelet count after 7 d. Steadystate thrombocytopoiesis was achieved by 3–4 weeks after the onset of thrombocytopenia.

Platelet dense granule ADP and ATP decreased significantly from 3.89±0.20 and 2.33±0.25 μmol/1011 platelets respectively at baseline to 2.17±0.37 and 1.68±0.37 μmol/1011 platelets respectively after 7 d (P<0.001 in both cases) and normalization was achieved only after 4 weeks. By contrast, the mean platelet volume and platelet content of β‐TG and PF4 did not change significantly throughout the course of study (P>0.1 in both cases). Platelet function, assessed by platelet aggregation ex vivo, demonstrated that platelet function was not impaired despite the significant decrease in dense granule ADP. We conclude that a selective, temporal reduction in platelet dense granule adenine nucleotides reflects changes in the thrombocytopoietic control mechanism secondary to induction of acute thrombocytopenia.
A new congenital dyserythropoietic anaemia
Authors: Ohisalo Jorma J. , Viitala Juha , Lintula Riitta , Ruutu Tapani
Published: March 2008

Summary A new dominantly inherited dyserythropoietic anaemia is described. In the bone marrow, many of the nonspecific morphological characteristics described in congenital dyserythropoietic anaemias were seen; however, dysmorphic cells were rare. The acidified serum test was positive with one out of 17 sera tested; the negative sera included two that had haemolysed HEMPAS erythrocytes in the acid Ham test. Anti‐i‐agglutination was negative. No aberrations of red cell membrane protein glycosylation were observed. Serum cholesterol was low. Bilirubin conjugation was deficient but icterus was resolved by treatment with phenobarbital.
Neutrophil function in patients on continuous ambulatory peritoneal dialysis
Authors: Harvey D. M. , Sheppard K. J. , Morgan A. G. , Fletcher J.
Published: March 2008

Summary Frequent and recurrent episodes of peritonitis are a major cause of morbidity in patients on continuous ambulatory peritoneal dialysis (CAPD). One factor contributing to this problem may be an abnormality of neutrophil function in these patients. We have therefore quantified phagocytosis and killing by circulating and peritoneal neu‐trophils from patients on CAPD with and without peritonitis. Circulating neutrophils from uninfected patients showed reduced phagocytosis of both Staphylococcus epidermidis and Candida guilliermondii because of an opsonic defect in CAPD serum and because of a defect of the neutrophils themselves.

In contrast, phagocytosis by circulating and peritoneal neutrophils from patients with peritonitis was normal. Intracellular killing of C. guilliermondii was normal in all groups of neutrophils but killing of S. epidermidis, the organism most commonly isolated in CAPD peritonitis, was reduced. The possible mechanisms for the enhanced neutrophil activity seen in peritonitis, and for the decreased killing of S. epidermidis in contrast to normal killing of C. guilliermondii are discussed. A defect in killing of S. epidermidis may explain why peritonitis caused by this organism can be difficult to erradicate.
Long‐term stability of international reference preparations for thromboplastins
Authors: Van Den Besselaar A. M. H. P. , Hermans J. , Beeser H. , Loeliger E. A.
Published: March 2008

Summary Three certified reference materials for thromboplastins are available from the Community Bureau of Reference (BCR) of the European Commission for calibration of commercial thromboplastins used for control of oral anticoagulant therapy. The long‐term stability of these reference materials has been monitored by two independent laboratories, using deep‐frozen and lyophilized plasma samples. Pro‐thrombin times and prothrombin time ratios measured on 19 occasions in the period 1981–86 have been analysed for trend with time. Although significant trends of prothrombin time and ratio (P<0.05) were observed, a consistent pattern of trends could not be recognized. The significant trends of prothrombin time and prothrombin time ratio are most probably due to changes in local laboratory conditions. There is no indication that the reference materials have deteriorated since the beginning of the study. It is recommended that long‐term stability monitoring of thromboplastins be performed by at least two laboratories simultaneously.
Impairment of primary haemostasis by low molecular weight heparins in rats
Authors: Borowska A. , Lauri D. , Maggi A. , Dejana E. , Gaetano G. de , Donati M. B. , Pangrazzi J.
Published: March 2008

SummaryDifferent low molecular weight (LMW) heparins were tested on primary haemostasis in rats. Four preparations were studied: one was devoid of any effect on the bleeding time, while the other three prolonged the bleeding time to varying extents. As a consequence we studied the effect of these heparins on platelet aggregation. The fractions which prolonged the bleeding time, also inhibited theex vivoandin vitroplatelet aggregation, whereas the one devoid of any effect on the bleeding time did not affect platelet aggregation. Similar results were obtained using both platelet‐rich plasma (PRP) and gel‐filtered platelets. Thein vitroresponse of platelets to aggregating agents may offer a parameter to detect the presence of ‘bleeding factor(s)’in some LMW heparin preparations.
Endogenous erythroid clones (EEC) in polycythaemia and their relationship to diagnosis and the response to treatment
Authors: Reid C. D. L. , Fidler J. , Kirk A.
Published: March 2008

Summary The growth in culture of circulating erythroid progenitors (BFU‐e) from 34 patients with erythrocytosis was evaluated together with the clinical and bone marrow features at presentation and the subsequent response to treatment. Clonal erythroid growth in vitro in the absence of added erythropoietin (EEC) was observed in 17/18 patients with clinically unequivocal polycythaemia vera and in these patients bone marrow morphology was also abnormal. EEC were also present in three out of five patients with only a single minor criterion of that disease but none of the 11 patients without major or minor criteria or evidence of secondary polycythaemia were EEC positive. This group had a very low incidence of bone marrow abnormalities, was probably heterogenous in nature and included two patients with elevated serum immunoreactive erythropoietin of unknown cause.

The reduction in haematocrit following treatment was assessed in EEC positive and negative patients and was found to be inferior in those lacking EEC. Thus both the poor therapeutic response and the low incidence of bone marrow abnormalities in patients lacking any other clinical features of polycythaemia vera correlated closely with the absence of EEC in in vitro culture. Myeloproliferative disorder may therefore be an unlikely cause of the erythrocytosis in such individuals.
Differential phthalocyanine photosensitization of acute myeloblastic leukaemia progenitor cells: a potential purging technique for autologous bone marrow transplantation
Authors: Singer C. R. J. , Linch D. C. , Bown S. G. , Huehns E. R. , Goldstone A. H.
Published: March 2008

Summary The potential value of sutphonated aluminium phthalocyanine (AISPc) as a purging agent for bone marrow autografts in acute myeloblastic leukaemia (AML) has been studied using in vitro clonogenic assays for normal (GM‐CFC) and leukaemic (AML‐CFC) progenitor cells. In nine out of 13 cases, the leukaemic blasts were found to be highly sensitive to AISPc. In six of the sensitive cases clonogenic assays revealed that only 2.1% of AML progenitor cells survived AISPc treatment under conditions which permitted a GMCFC recovery of 60.11%. AISPc photosensitization was also shown to selectively eliminate the leukaemic cell line K562 from an in vitro model of minimal residual disease. Thus photosensitization using AISPc may be an effective method of purging marrow autografts in some cases of AML. Evaluation of the sensitivity of AML clonogenic cells at diagnosis may identify those patients in whom AISPc photo‐purging may be of benefit at the time of an autologous bone marrow transplant.
Haemoglobin M‐Hyde Park associated with polyagglutinable red blood cells in a South African family
Authors: Bird A. R. , Kent P. , Moores P. P. , Elliott T.
Published: March 2008

Summary Twelve of 35 members tested in a large ethnically‐mixed South African family were found to have both haemoglobin M type Hyde Park and persistent polyagglutinable red blood cells. The characteristics of the polyagglutination have not been recorded previously. The cells of affected family members were not agglutinated by Arachis hypogea, Dolichos biflorus or Salvia sclarea, but were agglutinated weakly by Salvia horminum and BSII (GSII) and reacted strongly with Glycine soja and Sophora japonica lectins. BSI (GSI) lectin agglutinated the group A but not the group O cells. The N and MN cells were agglutinated more strongly than normal by Vicia graminea, other anti‐N lectins and human anti‐N but the M and MN cells reacted as expected with human anti‐M. The name ‘Hyde Park’is provisionally suggested for this type of polyagglutination. although it appears unlikely that the evidently complete association between the polyagglutination and the variant haemoglobin is the result of a single genetic mutation. More likely, the connection has a post‐genetic origin, perhaps showing that bonds, possibly affected adversely by precocious senescence, normally occur between the haemoglobin and α‐sialoglyco‐protein molecules in red blood cells.
Refined chromosome study helps define prognostic subgroups in most patients with primary myelodysplastic syndrome and acute myelogenous leukaemia
Authors: Yunis Jorge J. , Lobell Michael , Arnesen Mark A. , Oken Martin M. , Mayer Mary G. , Rydell Robert E. , Brunning Richard D.
Published: March 2008

Based on a 6·1/2‐year study of 284 consecutive adult patients with primary myelodysplastic syndrome (MDS) and and acute myelogenous leukaemia (AML), we have found that refined chromosome analysis can be used as an independent prognostic indicator in the great majority of patients with MDS and AML. In MDS, the FAB subtype was also found to have prognostic value and this was enhanced when the chromosomal findings were taken into consideration. In AML, the age of the patient correlated more closely with the chromosomal changes in predicting prognosis in most patients than did the FAB classification.

Previously we reported that refined chromosome analysis of bone marrow specimens from 161 adult patients with primary or non‐therapy related MDS and AML identified three prognostic chromosomal categories in each disease, representing 40% of all patients (Yunis et al, 1984, 1986). By extending our study to 284 patients, as well as a longer follow‐up, it was possible to determine the prognostic implications of two additional chromosomal categories in MDS and five in AML. Since 73% of all patients are now represented in well‐defined chromosomal subgroups with prognostic significance, refined chromosome analysis emerges as a tool that could have considerable impact in clinical decision making and in the development of treatment protocols.
Use of minisatellite DNA probes for recognition and characterization of relapse after allogeneic bone marrow transplantation
Authors: Min Gao Ling , Hibbin Jill , Arthur Christopher , Apperley Jane , Jeffreys Alec , Goldman John
Published: March 2008

Restriction fragment length polymorphisms can be used to distinguish blood and marrow cells from close relatives. We used two probes that recognize a series of dispersed and highly polymorphic tandem‐repetitive mini‐satellite regions in the human genome that can be detected via a shared 10–15 base pair core sequence similar to the generalized recombination sequence (χ) of E. coli. We have studied the resulting individual‐specific DNA fingerprints in 15 patients before and after allogeneic bone marrow transplantation performed for chronic myeloid leukaemia and in two patients transplanted for acute leukaemia. Early engraftment could be demonstrated at 3 weeks post‐transplant based on the recognition of cells of donor origin. One patient who failed to engraft had only recipient type marrow cells 3 months post‐transplant. Nine patients who relapsed after transplantation had only cells of recipient origin. In one patient who relapsed after transplantation with T‐cell depleted donor marrow, fractionation studies showed that his T‐cells at relapse were of recipient origin. We conclude that these minisatellite probes are valuable for characterizing the origin of different cell populations after marrow transplantation and could be useful for characterizing relapse when donor and recipient are of the same sex.
Postnatal changes in plasma burst‐promoting activity (BPA) levels in preterm infants
Authors: Iwai Yoshitaka , Nakato Hidekazu , Kim Kwang‐Myong , Tanaka Mayumi , Yoshimura Tomohide , Mayumi Mitsufumi , Mikawa Haruki
Published: March 2008

Summary. Two‐stage cell culture assays were used to determine burst‐promoting activity (BPA) levels in the plasma of untransfused premature infants during the early anaemic period. The plasma BPA levels decreased during the early phase of the postnatal fall in haemoglobin and the mean plasma BPA levels at 2 and 4 weeks of age were significantly lower (24 ± 9% and 20 ± 14%) than those of normal adults (53 ± 17%). After 8 weeks of age, plasma BPA levels increased markedly in correlation with the recovery of erythropoiesis. Inhibitors of erythroid colony growth were not significantly elevated in the plasma of premature infants. These results suggest that BPA may act as a regulator of erythropoiesis in preterm infants along with erythropoietin.
Bone marrow transplantation for myelodysplastic syndromes
Authors: Bélanger Robert , Gyger Martin , Perreault Claude , Bonny Yvette , St‐Louis Jean
Published: March 2008

Summary. Eight patients with myelodysplastic syndromes (MDS) were treated with bone marrow transplantation (BMT). Median age was 34.5 years and ranged between 3 and 45. FAB diagnosis was refractory anaemia (RA) in three, RA with excess of blasts (RAEB) in four and RAEB in transformation (RAEB‐t) in one case. Four patients were prepared with cyclophosphamide and total body irradiation whilst the other four received busulphan and cyclophosphamide. Engraftment was documented in seven of eight patients. Two patients died from complications related to the procedure. One had early veno‐occlusive disease of the liver whilst the other died 46 months after BMT from pulmonary fibrosis. One patient died from recurrent disease 11 months after BMT. Five patients are alive and in complete remission 9–35 months post‐transplantation. Four of these patients have a Karnofsky score ≤ 90%. These results suggest that BMT can induce prolonged disease‐free survival in patients under 50 years of age. If a compatible donor is available, marrow transplantation should be seriously considered in the treatment of MDS.
Childhood monosomy 7 revisited
Authors: Evans J. P. M. , Czepulkowski B. , Gibbons B. , Swansbury G. J. , Chessells J. M.
Published: March 2008

Summary. Monosomy 7 is found in acute myeloid leukaemia (AML) and myelodysplasia and is characteristic of a rare chronic myeloproliferative disease (MPD) of young children. We have seen 16 children with monosomy 7. Their clinical features and response to treatment are discussed.

Monosomy 7 diseases appear to have a particularly poor prognosis. The AML is often resistant to treatment and relapse is common. Children with chronic MPD die of bone marrow failure or evolve to AML or myelofibrosis. We have treated these children intensively with combination chemotherapy and allogeneic bone marrow transplantation. Four children with MPD received supportive care and low dose chemotherapy alone. They all died, surviving between 4 months and 4 years. Six children with MPD received intensive chemotherapy: three remitted, one relapsing after 9 months, the others remaining in remission at 18 months and 3 years. One child with MPD has undergone successful BMT and survives 71/2 years after presentation.

Remission was achieved in three of four cases of AML. They all relapsed within 9 months. Bone marrow transplantation was successful in one child with myelofibrosis.

Intensive chemotherapy and early bone marrow transplantation is likely to offer these children their best chance of survival.
Biochemical markers in multiple myeloma: a multivariate analysis
Authors: Simonsson B. , Källander C. F. R. , Brenning G. , Killander A. , Gronowitz J. S. , Bergström R. , Åhre A.
Published: March 2008

Summary.The analysis of individual biochemical and clinical variables in 121 patients with multiple myeloma showed that serum β2‐microglobulin (S‐β2m) had the most significant relation to survival. Other variables such as serum thymidine kinase (S‐TK), serum lactate dehydrogenase (S‐LDH), Screatinine, haemoglobin (Hb), ESR, S‐albumin, age and clinical stage were also significant. No such relationship was found with M‐component, presence of light chains in urine, type of secreted immunoglobulin or S‐calcium. The exclusion of clinical stage in the first multivariate analysis resulted in a model consisting of S‐β2m, age and S‐TK, none of the other variables gave additional information. When in the second multivariate analysis the basic variables involved in staging procedure were excluded and clinical stage included, stage III, but not stage II, was found to give additional information to the model described above.

Individual analysis of the variables showed that Hb had the most significant relation to effect of initial therapy. Other significant variables were S‐TK, S‐β2m and age. When using the multivariate approach, Hb alone was found to contain all the relevant information.
Alterations in size, shape and osmotic behaviour of red cells after splenectomy: a study of their age dependence
Authors: de Haan L. D. , Werre J. M. , Th. Ruben A. M. , Huls H. A. , de Gier J. , Staal G. E. J.
Published: March 2008

Summary. In whole blood from splenectomized subjects (n = 20), red cells showed a significant increase of mean surface area (MSA), mean cell volume (MCV), MSA/MCV‐ratio and osmotic resistance, with the mean cell haemoglobin concentration (MCHC) being decreased. Studies on red cell populations of different cell age revealed that the increase of MSA affects younger and older cells, whereas the increase of MCV can mainly be ascribed to young cells with low density. The increased osmotic resistance is mainly determined by older cells due to a more favourable MSA/MCV‐ratio. Shortly after splenectomy (n= 5) the MSA of younger and older cells increased, whereas the increase of MCV affected only young cells with a lowered density; moreover, the MSA/MCV‐ratio increased in older cells in particular, resulting in a relatively greater increase of osmotic resistance. An impaired maturation of the reticulocyte may underlie the initial increase of MSA and MCV of young cells, but the present results contradict the current view that delayed maturation explains the changes in morphology and osmotic resistance of asplenic red cells.
Distribution of the CD45R antigen in the maturation of lymphoid and myeloid series: the CD45R negative phenotype is a constant finding in T CD4 positive lymphoproliferative disorders
Authors: Huelin C. , Gonzalez M. , Pedrinaci S. , de la Higuera B. , Piris M. A. , Miguel J. San , Ruiz‐Cabello F. , Garrido F.
Published: March 2008

Summary. We have compared the expression of the common leucocyte antigen (CD45) and the restricted leucocyte antigen (CD45R) on normal haematopoietic cells, cell lines, and a total number of 136 cases of myeloid and lymphoid proliferative syndromes. CD45, the conventional leucocyte antigen, presents a generalized distribution along the lymphoid and myeloid maturation pathway with the exception of some myelomas and pre‐B leukaemias. In contrast, the expression of the CD45R determinant is more limited. Although it is found in the majority of the differentiation stages of B cells and monocytes, it is present only in the early stages of myeloid differentiation. On T cells it is expressed on mature thymocytes and in the majority of CD8+ lymphocytes and a subset of CD4+ cells on peripheral blood. Finally, our results also indicated that CD4+ T lymphoproliferative syndromes are derived from the CD4+ CD45R‐ subset (20/20 cases).
Decreased expression of the common acute lymphoblastic leukaemia antigen (CALLA/CD10) on neutrophils from patients with thermal injury
Authors: McCormack Robert T. , Nelson Robert D. , Solem Lynn D. , LeBien Tucker W.
Published: March 2008

Summary. The common acute lymphoblastic leukaemia antigen (CALLA/CD10) is a normal component of the circulating neutrophil cell surface membrane. In order to examine the potential functional significance of CALLA/CD10 we analysed the expression of this molecule on neutrophils isolated from thermal injury patients, since these patients have a well‐documented constellation of neutrophil defects affecting their microbicidal functions. Expression of neutrophil CALLA/CD10 was monitored by indirect immunofluorescence and flow cytometry. We observed that CALLA/CD10 expression was quantitatively reduced on burn patient neutrophils, compared to healthy donors (P < 0.001). In contrast, burn patient neutrophils expressed normal levels of class I HLA molecules and the C3bi receptor. Reduced expression of CALLA/CD10 was not associated with neutrophil activation or exposure to plasma ‘factor(s)’in vivo. Analysis of normal bone marrow neutrophils by cell sorting indicated that expression of CALLA/CD10 occurs late in neutrophil maturation, since 25% of polymorphonucleated bone marrow neutrophils did not express cell surface CALLA/CD10. Attempts to examine the chemotactic responses of CALLA/CD10 positive and negative neutrophils from burn patients were hampered by previous exposure of these cells to chemoattractants in vivo. Collectively, our findings suggest that burn patient peripheral blood neutrophils may be deficient in CALLA/CD10 due to insufficient maturation time in the bone marrow following thermal injury.
The role of ultrastructural cytochemistry and monoclonal antibodies in clarifying the nature of undifferentiated cells in acute leukaemia
Authors: Matutes Estela , de Oliveira Maria Pombo , Foroni Letizia , Morilla Ricardo , Catovsky Daniel
Published: March 2008

Summary. The nature of the cells in 21 cases of acute leukaemia with blasts which were undifferentiated by light microscopy criteria was investigated by immunophenotyping, ultrastructural cytochemistry and DNA analysis. Two groups of cases were recognized. Fourteen cases were negative with B and T lymphoid markers and expressed one or two myeloid antigens detected by the monoclonal antibodies (McAb) MCS2 (CD13) and MY9 (CD33). Peroxidase activity was demonstrated at ultrastructural level by the method of Roels on unfixed cells in eight out of 10 cases; rearrangement of the immunoglobulin (Ig) genes was demonstrated in one of the three cases investigated. These cases are proliferations of early, MO, myeloblasts which can only be recognized by immunological and ultrastructural cytochemical methods. The remaining seven cases revealed a complex phenotype with expression of myeloid and lymphoid antigens. Peroxidase activity was detected in blasts from two cases with rearrangement of the Ig‐heavy chain gene: in one of them the T cell receptor β and γ chain genes were also found in rearranged configuration. This group comprises cases of biphenotypic and mixed acute leukaemia which probably involve multipotent stem cells. This study demonstrates that the expression of myeloid antigens on blast cells parallels closely the presence of peroxidase activity and that lymphoid markers correlate with gene rearrangements at DNA level. Our findings are reassuring with respect to the specificity of the antimyeloid McAb for the diagnosis of cases which are unclassifiable by conventional methods.