bluuebunny/crossref_metadata_2025 huggingface dataset:
Double‐blind test of human urinary macrophage colony‐stimulating factor for allogeneic and syngeneic bone marrow transplantation: effectiveness of treatment and 2‐year follow‐up for relapse of leukaemia
Authors: Masaoka Tohru , Shibata Hirotoshi , Ohno Ryuzo , Katoh Shunichi , Harada Mine , Motoyoshi Kazuo , Takaku Fumimaro , Sakuma Akira
Published: March 2008

Summary A randomized, double‐blind placebo‐controlled phase III clinical trial was performed to study the effects of human urinary macrophage colony‐stimulating factor (hM‐CSF) after allogeneic and syngeneic bone marrow transplantation (BMT) in 60 hM‐CSP treated and 59 placebo control patients. HM‐CSF was administered at a daily dose of 2 ± 105 units/kg from day 1 to day 14 after RMT. Significant differences between hM‐CSF and control patients were found in the recovery time to greater than 0. 5 ± 109 granulocytes/1 and the survival rate during the initial 120 d without retransplantation. There was no difference in the incidence or grade of graft‐versus‐host disease (GVHD). There was no difference in the rate of leukaemic relapse at 24–36 months after BMT in patients with acute lymphocytic. acute non‐lymphocytic, or monocytic leukaemia. The results of this trial show that human M‐CSF improves the outcome of BMT without any influence on the occurrence of leukaemic relapse or GVHD.
Serum immunoreactive erythropoietin during pregnancy and in the early postpartum
Authors: Beguin Yves , Lipscei Gyorgyi , Oris Renée , Thoumsin Henri , Fillet Georges
Published: March 2008

Summary We studied 209 women during normal pregnancy, at delivery, or in the early postpartum. to determine whether erythropoietin (EPO) response was appropriate for the degree of anaemia. Serum immunoreactive EPO was measured in 74 nonpregnant women, including 33 normal subjects (16.4 ± 4.1 mU/ml) and 41 women with hypoplastic, haemolytic, dyserythropoietic, or iron‐deficient anaemia. An inverse linear relationship (R=‐0.88. P<0.0001) between log(EPO) and Hct was observed. Predicted EPO values were derived for each Hct and an O/P ratio of observed/predicted log(EPO) was calculated for each sample (1.00 ± 0.10, range 0.80–1.20). Serum EPO levels (mU/ml) were significantly higher during pregnancy (30 ± 16. n=142), at delivery (31 ± 16, n= 41), and on day 7 postpartum (37 ± 35, n= 26) than in normal women (P<0.001). EPO levels increased steadily from 18 ± 6 mU/ml in the first, to 26 ± 14 mU/ml in the second, and to 35 ± 18 mU/ml in the third trimester (P<0.0001). The O/P ratio was normal on day 7 postpartum (1.01 ± 0.16), at delivery (1.03 ± 0.16), and in the third trimester (0.96 ± 0.15), but was significantly reduced in the first two trimesters (0.88+0.15, P<0.001). A significant negative correlation between log(EPO) and Hct was lacking in the first two trimesters, was present but with a reduced slope during the third trimester and at delivery, and was normal postpartum. We conclude that EPO response to anaemia is impaired in early pregnancy, recovers in late pregnancy, and normalizes rapidly in the postpartum.
Teicoplanin in the treatment of Gram‐positive bacteraemia in neutropenic patients
Authors: Venditti A. Micozzi M. , Amadori S. , Pulsoni A. , And C. Tirindelli , Martino P.
Published: March 2008

Summary The increasing incidence of bacteraemia caused by Gram‐positive bacteria in neutropenic patients prompted the authors to evaluate, in a prospective trial, the role of teicoplanin in the treatment of this infection. Over a 15‐month period. 76 cases of bacteraemia (out of 265 evaluable episodes of fever) were observed at the Division of Haematology. University La Sapienza. Kome. Of the 76 cases studied, 46 (60%) were caused by Gram‐positive bacteria and 28 (37%) were caused by Gram‐negative bacilli. All febrile episodes were treated randomly and empirically with piperacillin plus amikacin with or without teicoplanin. Overall, 41 (54%) of the 76 cases of bacteraemia responded to the initial antibiotic regimen: with subsequent modifications the response rate rose to 96%.

In the treatment of Gram‐positive bacteraemia. first‐line administration of teicoplanin was found to be associated with early defervescence and with a significantly higher rate of success without modification of treatment (P < 0.01). Addition of teicoplanin as second‐line therapy produced a favourable outcome in 12 (70%) out of 17 cases of bacteraemia unresponsive to the initial piperacillin + amikacin regimen. No cases of Gram‐positive bacteraemia associated with septic shock or adult respiratory distress syndrome were observed in either treatment group. Only two late deaths were observed, and these occurred in patients with streptococcal septicaemia who were not receiving early teicoplanin.

The above data do not endorse the use of glycopeptide antibiotics in the early treatment of fever in neutropenic patients: rather, these compounds should be reserved for proven or presumed Gram‐positive infections which do not respond to initial beta‐lactam/aminoglycoside treatment.
A comparative efficacy and safety study of teicoplanin plus aztreonam versus gentamicin plus piperacillin in haematology oncology patients with clinically diagnosed septicaemia
Authors: Spencer K. C. , Taylor A. K. M. , Winfield D. A.
Published: March 2008

Summary Infections due to Gram‐positive bacteria, especially coagulase‐negative staphylococci. have been increasing in immunocompromised patients during the last 5 years because of an increased use of Hickman catheters and oral gut decontamination with quinolones. Teicoplanin. a new glycopeptide antibiotic. has a long plasma half‐life which allows once‐a‐day bolus administration, making it a‘user friendly’agent. A randomized comparative evaluation of teicoplanin plus aztreonam versus gentamicin plus piperacillin in leukaemic patients with a clinical diagnosis of septicaemia was undertaken. The objectives of this study were (1) to evaluate the efficacy and safety of teicoplanin and aztreonam in comparison to a 'standard antibiotic’regimen and (2) to assess the local and systemic tolerance of these drugs. Results of the study in more than 70 patients to date are presented, and the role of anti‐Gram‐positive antibiotics in the management of severe sepsis in immunocompromised patients is discussed.
Ceftazidime plus teicoplanin versus ceftazidime plus amikacin as empiric therapy for fever in cancer patients with granulocytopenia
Authors: Meunier F. , Vander Auwera P. , And M. Aoun , Bron D.
Published: March 2008

Summary In a prospective randomized study, 100 episodes of fever (>38°C) and granulocytopenia (<1000/μ1) in cancer patients were empirically treated with ceftazidime (2 g every 8 h) plus teicoplanin (400 mg every 8 h on day 1: 400 mg every day thereafter) or ceftazidime (2 g every 8 h) plus amikacin (500 mg every 8 h).

Bacteraemia. clinically documented infection and possible infection were documented in seven. 11 and 19 patients treated with ceftazidime plus teicoplanin and in 11, four and 17 patients treated with ceftazidime plus amikacin. Overall, the response rate was similar in the two groups of patients as was the need for treatment modifications and the rate of death.

For documented Gram‐positive bacteraemia, the response rate was 2/5 patients treated with ceftazidime plus teicoplanin and 2/7 with ceftazidime plus amikacin: for documented Gram‐negative bacteraemia. the response rate was 1/2 and 3/4 patients respectively. No breakthrough bacteraemia was observed. Tolerance was excellent. although renal toxicity (elevation of serum creatinine) was observed in three patients treated with ceftazidime plus teicoplanin and in none allocated to ceftazidime plus amikacin.
Role of glycopeptide antibiotics in the treatment of febrile neutropenic patients
Authors: Smith S. R. , Cheesbrough J. , Harding I. , Davies J. M.
Published: March 2008

Summary The exact timing of the introduction of the glycopeptide antibiotics teicoplanin and vancomycin in the management of the febrile neutropenic patient continues to be controversial. However, there are certain firm criteria now emerging.

Bacteraemia can be eradicated with a success rate approaching 100% in cases where the organism can be identified and shown to be sensitive.

Approximately 6 5% of cases of soft‐tissue infection, usually occurring with the use of a Hickman or equivalent indwelling catheter, are associated with the presence on culture of Gram‐positive organisms of presumed skin origin. Such infection is an indication for the early use of antibiotics with proven activity against coagulase‐negative staphylococci and diptheroids.

Resolution of fever in up to 50% of cases may result from using‘planned progressive therapy’: the introduction of specific Gram‐positive cover in patients who have failed to respond at 48–72 h to regimens such as a ureidopenicillin or a third‐generation cephalosporin with or without an amino glycoside. This approach reduces the number of patients who go on to receive empirical amphotericin B intravenously for presumed fungal infection.

Using teicoplanin or vancomycin as first‐line agents in the empirical treatment of first fever in febrile neutropenic patients is perhaps more controversial. Recent developments which include using quinolone‐based prophylaxis more widely and introducing cytokines to reduce the period of neutropenia may increase the likelihood that a neutropenic patient's febrile episode will be due to a Gram‐positive organism.

The dilemma of choosing broad‐spectrum monotherapy or targeted combination therapy in this situation remains unresolved. Current studies, however, should help to clarify this situation.

Finally, other current studies of teicoplanin and vancomycin as prophylactic agents administered either orally or systemically, may provide additional indications for their use in the neutropenic patient.
Simultaneous expression and regulation of G‐CSF and IL‐6 mRNA in adherent human monocytes and fibroblasts
Authors: Vellenga Edo , Van Der Vinne Bernard , De Wolf Joost Th. M. , Halie M. Ruud
Published: March 2008

Summary The regulation of granulocyte‐colony stimulating factor (G‐CSF) and interleukin‐6 (IL‐6) mRNA was studied in human adherent monocytes in response to the protein kinase C activator, oleolyl‐acetylglycerol (OAG), the calcium‐ionophore A23187 and the cyclic AMP elevating agents, dibutyryl c‐AMP (DBcAMP), cholera toxin and isobutyl‐methylxanthine (IBMX). G‐CSF and IL‐6 transcripts were simultaneously expressed in response to OAG, A23187, DBcAMP, IBMX and cholera toxin. However, the time course demonstrated a difference; a rapid induction by OAG and A23187 and a delayed pattern by cAMP elevating agents. In addition it appeared that the induction of CSFs by DBcAMP was independent of the adherence procedure or the presence of fetal bovine serum but could be counteracted by the simultaneous addition of H8, an inhibitor of the cAMP dependent kinases.

Finally, experiments were performed to study in how far comparable mechanisms operate in other cell types. Human fetal lung fibroblasts were stimulated with A23187. DBcAMP and OAG. All these agents induced simultaneous expression of G‐CSF and IL‐6 mRNA and secretion of proteins, indicating that different signalling pathways exist in both cell types which regulate the expression of both genes.
Thalidomide treatment for chronic graft‐versus‐host disease
Authors: Heney D. , Norfolk D. R. , Wheeldon J. , Bailey C. C. , Lewis I. J. , Barnard D. L.
Published: March 2008

Summary The treatment of chronic graft‐versus‐host disease (GVHD) may present a difficult therapeutic problem. We used thalidomide to treat six patients with severe chronic GVHD who failed to respond to standard immunosuppressive agents. Four of the six patients showed a clear response to thalidomide, with the fifth patient showing a partial response. The best results were seen in patients with chronic cutaneous GVHD. Two of the patients developed neurophysiological evidence of a peripheral neuropathy, associated with clinical signs in one patient. Measurement of thalidomide plasma levels and pharmacokinetic curves showed a significant inter‐patient variation. Peak plasma levels varied from 0·47 to 1·46 μg/ml. Thalidomide has a role to play in the management of chronic GVHD and further studies are needed.
Expression of the multidrug resistance gene product (P‐glycoprotein) in myelodysplasia is associated with a stem cell phenotype
Authors: List Alan F. , Spier Catherine M. , Cline Ann , Doll Donald C. , Garewal Harinder , Morgan Rodman , Sandberg Avery A.
Published: March 2008

Summary Previous studies have indicated relative resistance to chemotherapy in the myelodysplastic syndromes (MDS) and associated acute leukaemia. To determine if multidrug resistance may contribute to chemoresistance in these disorders, we studied bone marrow specimens for P‐glycoprotein expression (P‐GP) by immunocytochemical staining with monoclonal antibodies reactive with cytoplasmic (C219) or surface epitopes (MRK16) of P‐GP. Forty‐five case specimens from 43 patients were studied, including 32 cases of primary MDS, seven cases of acute myeloid leukaemia (AML) following MDS, and six therapy‐related haematological disorders. Cytogenetic analysis was available on 36 specimens. Two staining patterns were detected: (1) cytoplasm and plasma membrane, and (2) staining restricted primarily to the nuclear‐cytoplasmic junction. P‐GP was detected in seven (22%) cases of primary MDS, four (57%) cases of AML evolving from MDS, and five (83%) cases of therapy‐related haematological disorders. Expression of P‐GP was restricted to blasts and leukaemic monocytes, and was otherwise absent from terminally differentiated blood cells. Analysis of the relation between P‐GP expression and reactivity with the human progenitor cell antigen CD34, revealed a highly significant association (P= 0·001). P‐GP reactivity was distributed equally among normal and abnormal karyotypes and did not correlate with specific cytogenetic abnormalities. These findings indicate that multidrug resistance in MDS and karyotypically‐related haematological disorders is closely linked to a stem cell phenotype and may contribute to chemoresistance in these disorders.
Book Reviews
Authors: Stuart John , Scully Mike , Whittaker J. A. , Lane David , Machin Samuel J. , Testa N. G.
Published: March 2008

Book Reviews in This Article:

Practical Haematology, 7th edn. Edited by Sir John V. Dacie and S. M. Lewis.

The Biology of Antithrombins. By G. Sas.

William Dameshek and Frederick Gunz's ‘Leukaemia’, 5th edn. Edited by E. S. Henderson and T. A. Lister.

Fibrinogen 4. Current Basic and Clinical Aspects. Edited by M. Matsuda, S. Iwanaga, A. Takada and A. Henschen.

Disorders of Haemostasis, 2nd edn. Edited by O. D. Ratnoff and C. D. Forbes.

The Biology of Hematopoiesis. Edited by N. Dainiak, E. P. Cronkite, R. McCaffrey and R. K. Shadduck.
Myelodysplastic syndrome with increased marrow fibrosis: a distinct clinico‐pathological entity
Authors: Lambertenghi‐Deliliers Giorgio , Orazi Attilio , Luksch Roberto , Annaloro Claudio , Soligo Davide
Published: March 2008

Summary Seventeen cases of myelodysplastic syndrome (10 primary and seven secondary to previous radio‐chemotherapy), characterized by trilineage dysplasia, severe bone marrow fibrosis and a high number of megakaryocytes, are described. All of these patients had similar clinical and prognostic features consisting of pancytopenia, modest or absent visceral enlargement and poor survival. The use of CD61 antibodies, which recognize megakaryocytic cells at all stages of maturation, confirmed that these patients had a higher number of these cells than either normal subjects or patients affected by myelodysplastic syndrome (MDS) without fibrosis. Furthermore, primary and secondary MDS with fibrosis, although clinically and histopathologically similar, differed in terms of the number of megakaryoblasts which were significantly higher in primary forms (P<0·02). We conclude that MDS with fibrosis may represent a clinico‐pathological entity which needs to be distinguished from other MDS subtypes as well as from idiopathic myelofibrosis or malignant myelosclerosis.
Establishment and characterization of a B‐cell line derived from a patient with a myelodysplastic syndrome which expresses myelomonocytic and lymphoid markers
Authors: Bergamaschi Gaetano , Stella Carmelo Carlo , Cattoretti Giorgio , Invernizzi Rosangela , Maserati Emanuela , Nalli Giulio , Pedrazzoli Paolo , Peverali Fiorenzo A. , Valle Giuliano Della , Ascari Edoardo , Cazzola Mario
Published: March 2008

Summary We describe a novel continuous B‐cell line (PV‐90) derived from a patient with myelodysplastic syndrome (MDS) and originating from spontaneous infection with the Epstein‐Barr virus (EBV). The patient progressed to acute myeloblastic leukaemia (AML) 5 months after clinical onset of MDS. PV‐90 is of clonal origin as indicated by the presence of immunoglobulin (Ig) gene rearrangements, monoclonal surface immunoglobulins, and a single DNA restriction fragment corresponding to the EBV genomic termini. PV‐90 cells also express a number of myelomonocytic markers, including α‐naphthyl acetate esterase (ANAE), coagulation factor XIII, and CD68 antigen. Moreover, PV‐90 cells constitutively express the c‐fms proto‐oncogene mRNA as the patient's blast cells did. Whereas a trisomy 11 (+11) was found in the patient's bone marrow cells, PV‐90 cells had a normal karyotype initially, but at 4 months showed two different and independent chromosomal abnormalities: 90, XX, ‐Y, ‐Y, t(9;16) (q11;p13), and 90, XX, ‐Y, ‐Y, t(17;18) (p13;q21), the latter possibly involving the p53 (17, p13) and bcl‐2 (18, q21) proto‐oncogenes. The early development of these chromosomal aberrations is consistent with a genetic instability of PV‐90 cells. Expression of bi‐lineage markers and genetic instability may suggest that PV‐90 cells originated from transformation of a myelodysplastic progenitor cell capable of both myeloid and B‐cell differentiation. The PV‐90 cell line might be useful in a number of studies, including the possible role of c‐fms in cell differentiation, pathogenetic mechanisms of human preleukaemia and lineage promiscuity in acute leukaemia.
Effects of recombinant interleukin 4 on the growth and differentiation of blast progenitors stimulated with G‐CSF, GM‐CSF and IL‐3 from acute myeloblastic leukaemia patients
Authors: Imai Yasufumi , Tohda Shuji , Nagata Kaoru , Suzuki Toshiya , Nara Nobuo , Aoki Nobuo
Published: March 2008

Summary The effects of human recombinant interleukin 4 (rIL‐4) on the growth of leukaemic blast progenitors were investigated. Cells obtained from 20 acute myeloblastic leukaemia (AML) patients were evaluated using the blast colony assay in methylcellulose and suspension cultures. While rIL‐4 by itself did not show any colony stimulatory activity in the blast colony assay, it suppressed the blast colony formation in methylcellulose stimulated with G‐CSF, GM‐CSF or IL‐3 in 14 patients. In another six patients, rIL‐4 enhanced blast colony growth in four patients or did not show any significant effect with any CSF in two patients. In suspension cultures of 12 cases studied, the effects of rIL‐4 on the clonogenic cell recoveries were essentially similar to the results of the blast colony assay in each case. In three patients, rIL‐4 augmented the differentiation of the leukaemic cells to monocyte lineage. Further, the clinical outcome was significantly different between the patients whose blast progenitors were stimulated by rIL‐4 and the patients whose blast progenitors were suppressed by rIL‐4 (P<0.05): three out of four patients in the former group failed in achieving complete remission (CR), while 12 out of 14 patients in the latter group achieved CR. The results show that the effects of IL‐4 on leukaemic blast progenitors were diverse and the responsiveness to IL‐4 may be correlated with the prognoses of the patients.
BCNU treatment of marrow stromal monolayers reversibly alters haematopoiesis
Authors: Uhlman Dorothy L. , Verfaillie Catherine , Jones Roy B. , Luikart Sharon D.
Published: March 2008

Summary. The marrow stromal microenvironment is essential for maintaining normal haematopoiesis. Chemotherapy drugs, such as the nitrosoureas, may impair the ability of the stroma to support haematopoiesis. To assess the effects of 1,3‐bis‐(2‐chloroethyl)‐1‐nitrosourea (BCNU) on in vitro haematopoiesis, stromal monolayers were treated with BCNU, 5 μg/ml weekly for 3 weeks, then seeded 24 h after the third treatment with haematopoietic progenitors. Three weeks after seeding, total adherent and non‐adherent cell numbers were similar in treated and control flasks as were adherent granulocyte/macrophage colony forming cell (GM‐CFC) numbers. In contrast, non‐adherent GM‐CFC were significantly reduced in treated flasks, to 40–60% of controls. However, no reduction in non‐adherent GM‐CFC number was seen if seeding was delayed for 7 d following BCNU treatment, suggesting the effects are reversible. Conditioned media from treated and control monolayers, harvested at a time corresponding to the time of seeding, showed no difference in colony stimulating activity. In addition, extracts of bound growth factors from treated and control monolayers also showed no differences in colony stimulating activity. Thus, BCNU can alter haematopoiesis through a reversible effect on the marrow stroma and this effect does not appear to be mediated by a change in stromal growth factor production.
Lymphohaematopoietic chimaerism after bone marrow transplantation for chronic myeloid leukaemia: results of simultaneous cytogenetic analyses on T‐cell colonies, myeloid, and erythroid progenitor cells
Authors: Schmitz N. , Schlegelberger B. , Oberböster K. , Golchert K. , Suttorp M. , Löffler H.
Published: March 2008

Summary. Various lymphohaematopoietic compartments represented by cells from T‐cell colonies, myeloid progenitor cells (CFU‐GM), erythroid progenitor cells (BFU‐E), and bone marrow after short‐term culture (BM) have simultaneously been analysed in 15 patients receiving 17 bone marrow transplants for Philadelphia chromosome (Ph) positive chronic myeloid leukaemia (CML) or acute lymphoblastic leukaemia (ALL). The marrow grafts were not T‐cell depleted. Ten patients without relapse did not show any myeloid cells of host origin until their last follow‐up or until death. However, in four of these patients single lymphoid host cells not carrying the Ph chromosome were found after BMT without clinical consequences. In patients with cytogenetic or haematological relapse Ph positive metaphases were first detected in any of the progenitor cell compartments along with residual donor cells in two of three patients. BM became Ph positive after various time intervals. Another patient with CML became Ph positive in all compartments investigated at the same time. The only patient with Ph positive ALL remained completely Ph negative also when haematological and clinical relapse was evident. All patients with relapse exhibited complex clonal and non‐clonal chromosomal aberrations at the time of recurrence of the Ph chromosome. Such abnormalities not identical to those usually found with evolution of the disease and preferentially occurring in progenitor cells preceded the reappearance of Ph positive metaphases in one of our patients.
Differential dose‐related haematological effects of GM‐CSF in pancytopenia: evidence supporting the advantage of low‐ over high‐dose administration in selected patients
Authors: Kurzrock Razelle , Talpaz Moshe , Gomez Jesus A. , Estey Elihu H. , O'Brien Susan , Hirsch‐Ginsberg Cheryl , Koller Charles , Freireich Emil J. , Gutterman Jordan U.
Published: March 2008

Summary. Granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) is a multifunctional haematopoietin which can promote production of several blood cell lineages, though the predominant target cells are neutrophils, monocytes, and their precursors. Occasional undesirable clinical effects include eosinophilia, an increase in blasts, or thrombocytopenia. Here, we describe four patients who were treated with GM‐CSF, at subcutaneous doses significantly lower than are conventional, and experienced an unusual response pattern. Three patients had severe pancytopenia associated with chronic lymphocytic leukaemia (CLL) or myelodysplastic syndrome (MDS) and exhibited an unexpected switch in the responsive lineage on high‐ versus very low‐dose therapy. The two CLL patients developed marked eosinophilia (up to 10·0 x 109 cells/l) without an increase in neutrophils on 125–300 μg/m2/d of GM‐CSF. In contrast, when the dose was lowered to 10 μg/m2/d, the neutrophils rose to physiological levels, without significant eosinophilia. The MDS patient showed a rapid rise in peripheral blasts (baseline level = 0; post‐therapy level = 5·0 x 109/l), without a change in other cell types, when receiving 60 μg/m2/d of GM‐CSF. After GM‐CSF was held, blasts returned to baseline levels; reinstituting therapy at the very low dose of 6 μg/m2/d was followed by an increase in platelet counts from 50 to 185 x 109/l with only a minor increase in blasts. The fourth patient, who suffered from severe aplastic anaemia complicated by recurrent gastrointestinal haemorrhage, was only treated with the low‐dose regimen. He showed a predominant platelet effect with counts rising from 9 to 169 x 109/l. Very low‐dose GM‐CSF therapy was devoid of constitutional side effects. The biological implications of these GM‐CSF responses are discussed. Our results indicate that, in some patients, GM‐CSF may stimulate different target cells depending on the dose. Therefore, in contrast to the results of administration of many classical drugs, there may not always be a direct relationship between the amount of GM‐CSF given and the optimal effect.
HPA‐5b (Bra) neonatal alloimmune thrombocytopenia: clinical and immunological analysis of 39 cases
Authors: Kaplan C. , Morel‐Kopp M. C. , Kroll H. , Kiefel V. , Schlegel N. , Chesnel N. , Mueller‐Eckhardt C.
Published: March 2008

Summary. Maternal alloimmunization against fetal platelets can cause fetal and neonatal thrombocytopenia (NAIT). The HPA‐1a (PIA1, Zwa) antigen is by far the most common antigen implicated in NAIT. However, today another antigen often linked with that affection is HPA‐5b (Bra). This is a report of 39 cases of NAIT involving the HPA‐5b antigen. Thrombocytopenia may be of grave consequence. Three infants developed intracerebral haemorrhages (ICH). Of these, one died presumably as a consequence of ICH. Central nervous system (CNS) sequelae in the neonatal period was observed in two children. The potential hazards of death or disabling neurologic sequelae following intracerebral haemorrhage call for rapid and reliable diagnosis and effective therapy. Because there is high risk that subsequent pregnancies might be also affected by NAIT, the mothers of a previously affected child should be managed similarly to the HPA‐1b mothers (PIA2, Zwb). The antenatal diagnosis of thrombocytopenia should be made and if necessary the in utero therapy instituted.
Abnormal neutrophil adhesion in sickle cell anaemia and crisis: relationship to blood rheology
Authors: Boghossian S. H. , Nash G. , Dormandy J. , Bevan D. H.
Published: March 2008

Summary. Defects in neutrophil adhesion and migration may contribute to the susceptibility to infection seen in sickle cell anaemia (SCA). These dynamic defects may be influenced by abnormalities in blood rheology found in this disorder. A whole blood model was used to study neutrophil adhesion in SCA patients: neutrophil adhesion to protein coated glass was quantitated by measuring the rate of disappearance of neutrophils from heparinized whole blood circulating through a perfusion chamber. Twenty‐three adult patients (Hb SS) were studied in asymptomatic steady state, of whom nine were also studied during pain crisis, both before and 4–7 d after conventional therapy. Red cell and granulocyte filter ability and whole blood and plasma viscosity were also measured. The half‐time for disappearance from the perfusion system (t1/2) of neutrophils from patients in the steady‐state was 93·5 ± 8·4 min, compared to 49·1 ± 2·8 min in normal age‐matched controls (P=0·001). In crisis, t1/2 was further prolonged to 170·0 ± 16·1 min (P= 0·01 v. steady state). After therapy, t1/2 decreased to 57·0 ± 4·5 min (P=0·001 v. pre‐therapy state and P= 0·009 v. steady state) and was comparable to Hb AA controls. These findings reveal a neutrophil adhesion defect in SCA which worsens in crisis but is corrected following supportive therapy.

Red cell filterability (expressed as average resistance to flow and pore‐clogging particles) and white cell filterability (expressed as pore‐clogging particles) were also abnormal in SCA and were found to correlate with neutrophil adhesion. Plasma viscosity also correlated with adhesion t1/2.

The defect appears to be related to abnormal blood flow properties in SCA but the rheological factors cannot fully explain either the steady‐state defect or the marked changes in neutrophil adhesion during crisis.
Autocrine and/or paracrine mechanism operate during the growth of human bone marrow fibroblasts
Authors: Kimura Akiro , Katoh Osamu , Hyodo Hideo , Kusumi Shizuyo , Kuramoto Atsushi
Published: March 2008

Summary The growth of human marrow fibroblasts was studied at various plating cell densities. The growth rate of fibroblasts cultured with human plasma derived serum (PDS) increased depending on the cell density plated, whereas fibroblasts cultured with human serum proliferated almost independent of the cell density. Conditioned medium obtained from the culture with PDS at high cell density enhanced the growth of fibroblasts plated at low density, as well as that of fibroblast colony forming cells. Characterization of this conditioned medium showed heat unstable and acid stable peptide factor(s) with high molecular weight (∼106) may be responsible for this activity. These results suggest that autocrine and/or paracrine mechanism can operate during the proliferation of marrow fibroblasts, a process thought to be involved in the progression of marrow fibrosis.
Relationship between post‐translational glycosylation and anticoagulant function of secretable recombinant mutants of human thrombomodulin
Authors: Koyama Takatoshi , Parkinson John F. , Aoki Nobuo , Bang Nils U. , Müller‐Berghaus Gert , Preissner Klaus T.
Published: March 2008

Summary Two glycoforms of a soluble mutant of recombinant human thrombomodulin (rec.TM) were used to identify critical N‐ and O‐linked glycans of the endothelial cell thrombin receptor. While N‐linked glycans were not found to be involved in any function of rec.TM, an acidic chondroitin sulphate‐like glycosaminoglycan (CSGAG) was found to be critical for all the direct anticoagulant functions of rec.TM, including inhibition of thrombin‐mediated platelet aggregation. A glycoform of rec.TM lacking CSGAG had very poor anticoagulant activity. Furthermore, the glycoform of rec.TM possessing CSGAG showed strong inhibition by and had high affinity for poly‐cationic basic proteins, whereas the CSGAG‐deficient rec.TM did not. Monoclonal antibody binding as well as lectin mapping of rec.TM with agglutinins identified sialic acid containing O‐linked glycans in both glycoforms additional to the CSGAG in high molecular weight rec.TM. These findings define important molecular interactions modulating the anticoagulant function of TM, which appear to be critically regulated by CSGAG, and also showed that the overall post‐translational glycosylation pattern of the two glycoforms was very similar except for the presence of CSGAG. The possibility exists that differently expressed glycoforms of TM may be crucial for the expression of endothelial cell‐related anticoagulant potential in different vascular beds.
Red cell ABO incompatibility and production of tumour necrosis factor‐alpha
Authors: Davenport R. D. , Strieter R. M. , Kunkel S. L.
Published: March 2008

Summary Tumour necrosis factor‐alpha (TNF) is a major mediator of diverse pathophysiological events similar to those of haemolytic transfusion reactions (HTR), such as fever, intravascular coagulation and organ failure. However, the possible role of TNF in HTR has not been investigated. We have constructed an in vitro whole blood model of HTR to examine whether TNF may be produced in red cell ABO incompatibility. TNF was observed in plasma, in a dose dependent manner, when ABO incompatible red cells were added, but not with compatible (group O) cells. Plasma TNF levels were maximal at 2 h, and declined to control levels by 24 h. Haemolysis of incompatible red cells was accompanied by TNF production. Immune haemolysis induced TNF gene expression by buffy coat leucocytes, as determined by Northern blot analysis. Heat inactivation of plasma abolished TNF production, whereas prior treatment with interferongamma augmented the response. These results demonstrate that a major cytokine is produced in response to red cell incompatibility, and suggest that TNF may play a role in the pathogenesis of haemolytic transfusion reactions.
Post consolidation therapy for adult patients with acute myeloid leukaemia
Authors: Ranson M. R. , Scarffe J. H. , Morgenstern G. R. , Chang J. , Anderson H. , Deakin D. P. , Oppenheim B. , Heron D. , Ryder D.
Published: March 2008

Summary One hundred and sixteen adult patients aged 14–73 with previously untreated acute myeloid leukaemia received induction and consolidation chemotherapy with daunorubicin, cytosine arabinoside and thioguanine. Two novel approaches to post consolidation therapy have been investigated. Patients aged 50 years or less who had no suitable matched allogeneic donor were considered for autologous bone marrow transplantation (BMT) using bone marrow which had been cultured in vitro for 14 d. Patients over the age of 50 years with normal bone marrow cellularity and peripheral blood count were treated with a single oral dose of busulphan 100 mg/m2 (without BMT rescue) 3 months following the completion of consolidation therapy. Eighty‐seven patients (75%) achieved a complete remission. Of 70 patients who completed consolidation therapy, 40 were aged ± 50 years and 30 were > 50 years. Forty‐three patients went on to receive post consolidation therapy in first CR (autologous BMT 12, allogeneic BMT 7, busulphan therapy 24). The event‐free survival at 4 years was 47% for autologous BMT, 34% for allogeneic BMT and 45% for busulphan‐treated patients. The survival for the older cohort of patients who received post consolidation therapy with single dose busulphan therapy was encouraging, and this agent should be considered for future post consolidation strategies.
Cytomegalovirus DNA in the sera of patients with cytomegalovirus pneumonia
Authors: Ishigaki Seishi , Takeda Mitsuaki , Kura Toshiro , Ban Noriyoshi , Saitoh Tadanori , Sakamaki Sumio , Watanabe Naoki , Kohgo Yutaka , Niitsu Yoshiro
Published: March 2008

Summary We attempted to detect cytomegalovirus DNA (CMV‐DNA) in the sera of four leukaemia patients who underwent an allogeneic bone marrow transplant (BMT), in six leukaemia patients who suffered from pneumonia and in 16 healthy subjects, using the polymerase chain reaction (PCR). Three of the four BMT patients subsequently developed CMV pneumonia. In two cases, CMV‐DNA was detected in the sera at about the time the pneumonia occurred, and the amount of DNA increased with disease progression. The serum of the third patient became positive for CMV‐DNA before he developed pneumonia. The fourth patient did not develop CMV pneumonia, but his urine became persistently positive for CMV‐DNA soon after the BMT, whereas the serum was negative. A relationship was found between the occurrence of pneumonia and the serum level of CMV‐DNA. CMV‐DNA was also detected in three of six pneumonia patients whose anti‐CMV IgM antibodies were elevated in the circulation. Sera from the 16 normal subjects were negative for CMV‐DNA, regardless of their being seropositive or seronegative for CMV.

While it had been previously thought that CMV did not exist in serum, we detected CMV‐DNA in serum by PCR in the active disease stage. Our results suggest that PCR would be useful for the early diagnosis of CMV pneumonia and in monitoring its course.
The rate of fibrinopeptide B release modulates the rate of clot formation: a study with an acquired inhibitor to fibrinopeptide B release
Authors: Nawarawong Weerasak , Wyshock Edward , Meloni Frank J. , Weitz Jeffrey , Schmaier Alvin H.
Published: March 2008

Summary An asymptomatic 50‐year‐old male with a gamma globulin paraprotein was found to have prolonged prothrombin time, activated partial thromboplastin time, and thrombin time but a normal reptilase time. The prolonged clotting times were not the result of a factor deficiency because they were not corrected by the addition of normal plasma. Instead, this patient had an antibody that delayed thrombin‐mediated fibrinopeptide B release thereby producing an apparent dysfibrinogenaemia. His isolated IgG prolonged the thrombin clotting time of both normal plasma and fibrinogen. Precincubation of his IgG with fibrinopeptide B, but not with fibrinopeptide A or thrombin, decreased its ability to prolong the thrombin clotting time. The patient's purified IgG but not control IgG delayed thrombin‐mediated fibrinopeptide B release from fibrinogen without affecting the release of fibrinopeptide A. These studies define a novel, clinically silent dysfibrinogenaemia due to an antibody that delays thrombin‐mediated fibrinopeptide B release from fibrinogen thereby markedly prolonging the clotting times.
Congenital dyserythropoietic anaemia with novel intra‐erythroblastic and intra‐erythrocytic inclusions
Authors: Wickramasinghe S. N. , Illum N. , Wimberley P. D.
Published: March 2008

Summary A hitherto undescribed form of congenital dyserythropoietic anaemia is reported. The patient was severely anaemic and hydropic at birth and is now 8 years old. She has a moderate normochromic normocytic anaemia, HbF level of 50%, reticulocyte count of 5‐12% and hyperbilirubinaemia. Bone marrow smears showed intense normoblastic erythroid hyperplasia with morphological evidence of dyserythropoiesis; the most common dysplastic features were basophilic stippling of polychromatic erythroblasts and of erythrocytes and marked abnormalities of nuclear shape in polychromatic erythroblasts. Electron microscope studies showed that some polychromatic erythroblasts and several erythrocytes contained inclusions which were rounded, elongated or irregular in outline or were doughnut‐shaped. These inclusions consisted of compact masses of tubules and saccules which may represent smooth endoplasmic reticulum together with Golgi cisternae. The ultrastructural studies also revealed peculiar membrane‐bound cylindrical structures in a rare late erythroblast, and phagocytosed erythroblasts within some macrophages. The technique of combined Feulgen micro‐spectrophotometry and 3H‐thymidine autoradiography demonstrated a pile‐up of early polychromatic erythroblasts in the G1 and G2 phases of the cell cycle, indicating a prolongation of, or an arrest at, these phases. Furthermore, nearly a quarter of all erythroblasts failed to incorporate 3H‐leucine into protein. Thus the anaemia appeared to be due to a combination of disordered erythroblast function, increased ineffectiveness of erythropoiesis and peripheral haemolysis. The primary defect may be an excessive synthesis or impaired degradation of intracytoplasmic membranes.
Effect of pharmacologic manipulation of protein kinase C by phorbol dibutyrate and bryostatin 1 on the clonogenic response of human granulocyte‐macrophage progenitors to recombinant GM‐CSF
Authors: McCrady Carl W. , Staniswalis Joan , Pettit George R. , Howe Craig , Grant Steven
Published: March 2008

Summary. The effects of pharmacologic manipulation of protein kinase C (PK‐C) activity on the response of committed human myeloid progenitor cells (CFU‐GM) to recombinant human granulocyte‐macrophage colony stimulating factor (rGM‐CSF) was assessed. Coadministration of the PK‐C activating agents, phorbol dibutyrate (PDBu) or bryostatin 1, with rGM‐CSF resulted in a dose‐dependent and, under some conditions, highly synergistic increase in the number of CFU‐GM. With optimal combinations, colony formation far exceeded that which could be obtained with high concentrations of rGM‐CSF alone. High concentrations of PDBu (e.g. ≥ 50 nM), but not bryostatin 1, completely inhibited the CFU‐GM response. These inhibitory effects could be reversed by bryostatin 1, but not by high concentrations of rGM‐CSF. Bryostatin 1 also potentiated colony formation in response to rGM‐CSF, and blocked the inhibitory effects of high concentrations of PDBu in bone marrow cells highly enriched for progenitors bearing the MY‐10 antigen. The increase in CFU‐GM induced by PDBu or bryostatin 1 was associated with little change in the morphologic type of colony observed. Continuous exposure of cells to the calcium ionophore, ionomycin (500 nM), reduced the number of granulocyte‐macrophage colonies, but produed little change in the concentration‐response of rGM‐CSF and PK‐C activating agents. Finally, the PK‐C inhibitors H‐7 and tamoxifen, when administered at concentrations exhibiting minimal inhibitory effects in the presence of rGM‐CSF alone, led to no change or small increases in the numbers of colonies formed in response to rGM‐CSF and bryostatin‐1, and a substantial increase in the number of colonies formed in the presence of rGM‐CSF and PDBu. These results suggest that PK‐C activation may play a complex role in regulating the response of normal myeloid progenitors to growth factors such as rGM‐CSF. They also raise the possibility that under some circumstances the phorbol ester PDBu may trigger events that inhibit the growth of myeloid progenitors, and that this process may be blocked by bryostatin 1.
Expression of triplicated and quadruplicated α globin genes in sheep
Authors: Vestri R. , Pieragostini E. , Yang F. , Gregorio P. di , Rando A. , Masina P.
Published: March 2008

Summary. In the sheep ααα globin gene haplotype, the three genes display from the 5′ to the 3′ end the percentage efficiencies of about 30:14:6, as indicated by the amounts of the three types of α chain produced in the ααα/ααα homozygotes. The 3′ gene in the αααα haplotype appears to have an efficiency around 1%, as suggested by analysis of one quadruple a homozygote. Moreover, the total outputs of the ααα as well as of the αααα haplotypes do not substantially differ from that of the common αα haplotype.
Sensitivity of erythroid progenitor colonies to erythropoietin in azidothymidine treated immunodeficient mice
Authors: Chow Fung‐Phing Rose , Sutton Patricia A. , Hamburger Anne W.
Published: March 2008

The anaemia induced by 3′‐azido‐3′dideoxythymidine (AZT) is poorly understood. We have used a murine model of AIDS, infection of female C57BL/6 mice with LP‐ BM5 murine leukaemia (MuLV) virus, to determine if AZT‐ induced anaemia is due, in part, to decreased responsiveness of erythropoietic precursors (BFU‐e) to erythropoietin (EPO). Mice in the early stage of LP‐BM5 MuLV disease were given AZT in their drinking water at 1·0 and 2·5 mg/ml. AZT produced anaemia in both groups, in a dose‐dependent fashion. Despite the anaemia, the number of splenic and bone marrow BFU‐e in AZT treated mice increased up to five‐fold over levels observed in infected untreated animals after 15 d of treatment. Colony formation by splenic and bone marrow BFUe was stimulated at lower concentrations of EPO in mice receiving AZT for 15 d than for infected, untreated mice. By day 30, sensitivity of both splenic and bone marrow BFU‐e of treated animals returned to that observed from cells of infected untreated animals. The mean plasma levels of EPO observed in AZT treated mice were appropriate for the degree of anaemia observed when compared with phenylhydrazine (PHZ) treated mice. The numbers of BFU‐e and the percent‐ age of bone marrow erythroblasts observed were comparable in AZT and PHZ treated mice with similar degrees of anaemia. However, reticulocytosis was inappropriate for the degree of anaemia observed in AZT treated infected mice. AZT‐induced peripheral anaemia in the face of increased numbers of BFU‐e and increased levels of plasma EPO suggest a lesion in terminal differentiation.
Absence of phosphatidylinositol (PI)‐linked proteins in a very early human multipotential haematopoietic marrow cell
Authors: Huang Andrew T. , Panella Timothy J. , Mold Nelda G. , Rosse Wendell F.
Published: March 2008

A very early human haematopoietic progenitor cell population which was negative for the major histocompatibility class II antigen (HLA‐DR) and positive for the CD34 (MY10) antigen was separated into two subsets according to the expression of decay‐accelerating factor (DAF) on the cell surface. Using immunoadherence, cell cycle analysis, and cell culture, we determined that there is a DAF− multipotential cell and a more differentiated DAF+ lineage specific progenitor cell existing in human bone marrow. The DAF+ subset was highly enriched for CFU‐GEMM, while the DAF+ subset contained only BFU‐E and CFU‐GM. The DAF− subset was approximately 0·03% and the DAF+ subset approximately 0·0·08% of the original bone marrow population. MIRL (membrane‐inhibitory of reactive lysis), another Pl‐linked protein, was not expressed on the DAF− population but was expressed on the DAF+ cells. These observations indicate that PI‐linked proteins are absent from the multipotential stem cell but are present on an early lineage specific cell. The absence of expression of PI‐linked proteins can be used to further isolate and characterize a very early multipotential haematopoietic progenitor cell population.
The effects of interferon‐α on the proliferation of CML progenitor cells in vitro are not related to the precise position of the M‐BCR breakpoint
Authors: Dowding C. , Guo A‐P. , Maisin D. , Gordon M. Y. , Goldman J. M.
Published: March 2008

We investigated the effects of brief (2 h) and continuous exposure to recombinant interferon‐α2a (rIFN‐α) on the proliferation of primitive (blast colony‐forming cells, Bl‐CFC) and committed myeloid progenitor cells (BFU‐E and GM‐CFC) derived from blood and bone marrow of patients with chronic myeloid leukaemia (CML) and normal subjects. In all three clonogenic assays, rIFN‐α suppressed colony formation in a dose‐dependent manner. No differences were detected in the proliferation of CML or normal Bl‐CFC and GM‐CFC exposed to rIFN‐α. Erythroid colony formation by normal, but not by CML BFU‐E, was inhibited by relatively low concentrations (100 U/ml) of rIFN‐α. However, in patients whose blood or marrow contained a mixture of Philadelphia chromosome (Ph)‐positive and Ph‐negative BFU‐E, cytogenetic analysis of individual erythroid colonies showed no differential inhibition by rIFN‐α. We found no difference in the sensitivity to rIFN‐α of GM‐CFC from patients whose leukaemic cells expressed BCR/ABL mRNA with the b2a2 junction and that of GM‐CFC from patients with the b3a2 mRNA. We conclude that (1) rIFN‐α does not have a significant leukaemia‐specific effect on the progenitor cells detected in these assays, and (2) the sensitivity of CML GM‐CFC to rIFN‐α is independent of the type of BCR/ABL message present in the cells. The clinical efficacy of rIFN‐α could be due to selective toxicity to cells not assayed in this study, to effects on accessory cells or to alterations induced in progenitor cell/stromal cell interactions.