bluuebunny/crossref_metadata_2025 huggingface dataset:
Association of inherited dysfibrinogenaemia and protein C deficiency in two unrelated families
Authors: Gandrille S. , Priollet P. , Capron L. , Roncato M. , Fiessinger J. N. , Aiach M.
Published: March 2008

Summary An inherited association of dysfibrinogenaemia and protein C deficiency was found in three members of the same family. The propositus was a 48‐year‐old man who suffered from severe and rapidly complicated atherosclerosis of the aorta and lower limbs arteries, which perhaps suggests that the association of these two molecular abnormalities may have enhanced the thrombotic process. The abnormal fibrinogen had a reduced ability to bind thrombin which may be thrombogenic.

We found the same inherited association of dysfibrinogenaemia and protein C deficiency in a patient with venous thrombosis. The functional abnormality of the fibrinogen, which could have been responsible for thrombosis, was delayed proteolysis by plasmin. Not only fibrinogen, but also fibrin clots were resistant to plasmic degradation. These observations raise two questions: (1) Is the association of a protein C deficiency with a dysfibrinogenaemia fortuitous or the result of a common mechanism? (2) Is there a link between an increased thrombotic tendency and either both of the defects of haemostasis that we have found, or only one of them?
Disturbed patterns of globin chain synthesis in childhood monosomy 7 myeloproliferative syndrome
Authors: Sheffer R. , Cividalli G. , Zaharan Y. , Tamari H. , Okon E.
Published: March 2008

Summary Two children with typical clinical and haematological features of monosomy 7 myeloproliferative syndrome are presented. Both children displayed decreased production of beta‐globin chains and unbalanced high alpha/non‐alpha synthetic ratios similar to those characteristic of homozygous β‐thalassaemia. These provide further evidence for the involvement of the erythroid line as part of the malignant clone, indicating neoplastic transformation of a pluripotential stem cell in this disease.
Complement is not activated in ABO‐haemolytic disease of the newborn
Authors: Brouwers H. A. A. , Overbeeke M. A. M. , Huiskes E. , Bos M. J. E. , Ouwehand W. H. , Engelfriet C. P.
Published: March 2008

Summary We studied the lysis in vitro of group A red cells by IgG anti‐A. IgG anti‐A, which strongly lysed A red cells from adults, did not lyse A red cells from cord blood, if fresh cord serum from a child with blood group AB was used as a source of complement. In cases of haemolytic disease of the newborn due to A‐O or B‐O antagonism with a positive direct antiglobulin test with anti‐IgG serum, the red cells did not react with anti‐complement sera. Apparently, complement is also not activated in vivo in case of A‐O haemolytic disease of the newborn.
Polynuclear iron complexes impair the function of polymorphonuclear granulocytes
Authors: Hoepelman I. M. , Jaarsma E. Y. , Verhoef J. , Marx J. J. M.
Published: March 2008

Summary In a previous study we showed that the phagocytic function of non‐stimulated PMN was impaired after incubation with high concentrations (200 μm) of polynuclear Fe(III), probably as a result of continuous generation of small amounts of superoxide and subsequent formation of hydroxyl radicals (Van Asbeck et al, 1984b). Because polynuclear Fe(III) complexes may not be available for biological reactions we have studied the effects of polynuclear and mononuclear iron(III) on the PMN. Fe(III) in its polynuclear form (Fe: citrate 1:1) was deleterious for the phagocytic function of PMN, while the mononuclear form (Fe:citrate 1:20) was not toxic. Binding affinity of polynuclear Fe(III) for PMN was higher than of mononuclear Fe(III), and a considerable amount of bound Fe(III) was found in the cytosolic fraction of non‐stimulated PMN. Limit dilution analysis of polynuclear complexes revealed that concentrations as low as 25 μm Fe(III) significantly impaired phagocytic function. The molecular weight of these complexes is similar to that of the non‐transferrin plasma iron found in the serum of patients with iron overload.

The toxic effects of small polynuclear non‐transferrin plasma Fe(III) complexes on PMN function may contribute to the development of infections in patients with iron overload.
Alkaline phosphatase positive precursors of adipocytes in the human bone marrow
Authors: Bianco P. , Costantini M. , Dearden L. C. , Bonucci E.
Published: March 2008

Summary Developing fat cells in the bone marrow of leukaemic patients treated with chemotherapy were found to be endowed with membrane‐bound alkaline phosphatase. Since alkaline phosphatase is a cytochemical marker of ‘reticular’cells, this observation provides cytochemical evidence that reticular cells may convert to adipocytes when marrow cellularity abruptly decreases.
Treatment of gold‐induced aplastic anaemia with immunosuppressive therapy
Authors: Doney Kristine , Storb Rainer , Buckner C. Dean , Thomas E. Donnall
Published: March 2008

Summary. Between June 1978 and August 1985 12 patients with gold‐induced aplastic anaemia were treated with immunosuppressive therapy. Median age was 38.5 years for the 11 females and one male. Treatment regimens varied, but all patients received 150–160 mg/kg horse anti‐human thymocyte globulin (ATG) over 4–10 d. Five patients had complete recovery of peripheral counts and one patient had a partial response. Of the six nonresponders one had a spontaneous recovery a year later and five died 6–67 d after beginning treatment. Seven patients are alive 4.3–8.9 years later. Six patients remain haematologically stable and one patient has developed a myelodysplastic syndrome. This experience confirms earlier reports of patients recovering marrow function after receiving immunosuppressive therapy for gold‐related aplasia.
Demonstration of three distinct immunological disorders on erythropoiesis in a patient with pure red cell aplasia and autoimmune haemolytic anaemia associated with thymoma
Authors: Taniguchi Shuichi , Shibuya Tsunefumi , Morioka Eiji , Okamura Takashi , Okamura Seiichi , Inaba† Shoichi , Niho Yoshiyuki
Published: March 2008

Summary. A patient with pure red cell aplasia (PRCA) and autoimmune haemolytic anaemia (AIHA), associated with a thymoma which had already been removed, was studied in order to investigate the pathogenesis of PRCA and AIHA. The autoantibody eluted from the surface of the patient's red blood cells (RBC) reacted with the large E antigen of the Rh complex. Immunoglobulin‐G (IgG) purified from the patient's serum suppressed CFU‐E and BFU‐E but not CFU‐GM colony formation in the presence of complement. This antibody was not adsorbed with large E antigen. T‐lymphocytes in the bone marrow suppressing autologous CFU‐E and BFU‐E colonies were demonstrated. Thus, three distinct immunological disorders on erythropoiesis were present in this patient with PRCA and AIHA associated with thymoma in a thymectomized state.
Enrichment of haemopoietic progenitor cells from the marrow of patients with myelodysplasia
Authors: Baines Paul , Masters Gillian S. , Lush Christopher , Jacobs Allan
Published: March 2008

Myeloid and erythroid progenitors from myelodysplastic marrows have been separated from accessory cell populations likely to influence their in‐vitro growth. Myeloid colony‐forming cells were enriched 23‐fold and erythroid progenitors 5‐7‐fold but, in both cases, retained their abnormal growth characteristics. After enrichment and removal of lymphocytes and monocytes, erythroid burst formation became markedly more dependent on the addition of 5637 bladder carcinoma conditioned medium as an exogenous source of haemopoietic growth factors. This suggests that lymphocytes and monocytes usually support erythropoiesis in cultures of myelodysplastic marrow and demonstrates that erythroid burst‐forming progenitor cells from myelodysplastic patients can respond to these populations in vitro. Haemopoietic failure in myelodysplasia appears to result from a defect within the preleukaemic clonogenic cell, rather than from an aberration in exogenous factors. The procedure outlined here can be used as a first step towards the isolation of these abnormal progenitors.
Polyclonal integration of HTLV‐I proviral DNA in lymphocytes from HTLV‐I seropositive individuals: an intermediate state between the healthy carrier state and smouldering ATL
Authors: Yamaguchi Kazunari , Kiyokawa Tetsuyuki , Nakada Kiyonobu , Yul Lee Song , Asou Norio , Ishii Toshinori , Sanada Isao , Seiki Motoharu , Yoshida Mitsuaki , Matutes Estela , Catovsky Daniel , Takatsuki Kiyoshi
Published: March 2008

We have studied 15 individuals (aged 14‐74 years) with antibodies to HTLV‐I in their serum and random integration of HTLV‐I proviral DNA in their peripheral blood lymphocytes. All but one of these patients suffered from a variety of non‐specific complaints which did not correspond to those of adult T‐cell leukaemia (ATL). All of them were born in Kyushu and Okinawa which are endemic areas for HTLV‐I infection; 25% of their family members were also seropositive for HTLV‐I. The only haematological abnormality in these patients was the presence of few atypical lymphoid cells in the peripheral blood. The CD4/CD8 ratios were normal but the proportion of Tac positive cells was slightly higher than normal.

These individuals with polyclonal integration of HTLV‐I proviral DNA seem to represent an intermediate state between smouldering ATL (monoclonal integration) and healthy HTLV‐I carriers (with antibodies but no detectable HTLV‐I proviral DNA). Patients with this intermediate state of HTLV‐I infection may be at risk to progress to ATL. The natural history of HTLV‐I infection in humans leading to the development of ATL is reviewed in the light of these new findings.
Reversal of drug‐resistance in multiple myeloma with verapamil
Authors: Durie Brian G. M. , Dalton William S.
Published: March 2008

In vitro testing with a new doxorubicin resistant myeloma cell line revealed the reversal of drug resistance with as little as 100 ng/ml of verapamil (a calcium channel blocker), a dose easily achievable in humans. A first patient with IgGk Myeloma is presented in whom resistance to VAD (vincristine/doxorubicin infusion plus dexamethasone) chemotherapy was reversed with the administration of verapamil. A subsequent clinical study has been initiated. The potential for further evaluation of calcium channel blockers in multiple myeloma is discussed.
Interleukin 2 production in bone marrow of normal individuals and patients associated with B‐cell chronic lymphocytic leukaemia
Authors: Rossi Jean‐François , Klein Bernard , Commes Therese , Jourdan Michel , Janbon Charles
Published: March 2008

T‐cells from patients with B‐cell chronic lymphocytic leukaemia (B‐CLL) have abnormal T4/T8 ratios and functions. Previously, we demonstrated that peripheral blood (PB) mononuclear cells from B‐CLL patients secrete significant amounts of interleukin 2 (IL2) with an apparent dysregulation of accessory cells controlling this production. In this study, IL2 production was investigated in PB and in bone marrow (BM) from patients with previously untreated B‐CLL, mostly in early stages of the disease, and compared to normal donors. A significant secretion was observed in both PB and BM from B‐CLL patients after stimulation by phyto‐haemagglutinin (PHA), with lower amounts in patients with nodular involvement of BM, as compared to interstitial or diffuse involvements. To explore the role of accessory cells in controlling IL2 production, we added phorbol ester or indomethacin to the culture system or irradiated the cells before culture. Phorbol ester significantly increased the IL2 secretion in both the PB and the BM of B‐CLL patients. The irradiation or the addition of indomethacin did not enhance the IL2 production in PB from B‐CLL patients. However, IL2 secretion increased in the BM cells from B‐CLL patients after addition of indomethacin or prior irradiation, in a similar way to that observed in PB and BM of normal controls, suggesting an apparent normal control of the IL2 production in BM from B‐CLL patients. In normal controls, we demonstrated that IL2 secretion per T‐cell from BM was 5·4‐fold greater than that from normal PB, suggesting a very efficient role of accessory cells controlling IL2 production in normal BM.
Platelet‐associated immunoglobulins G, A and M are secreted during platelet activation: normal levels but defective secretion in grey platelet syndrome
Authors: Pfueller Sharron L. , David Rosemary
Published: March 2008

Platelet‐associated (PA) IgG is known to be released from normal human platelets when they are stimulated by aggregating agents. We have studied whether PA‐IgA and PA‐IgM are also secreted during platelet activation or during blood collection and processing and whether their levels are related to those in serum. Processing of platelets from normal donors in the presence of secretion inhibitors prostaglandin E1 (PGE1) and theophylline increased levels of both surface and total PA‐immunoglobulins (Ig) in intact and lysed platelets respectively, with increases being significant for surface PA‐IgA and PA‐IgM and total PA‐IgM. About 50% of total PA‐IgM, 40% PA‐IgA and 20% PA‐IgG was detectable on intact platelets. All three PA‐Ig and PA‐albumin were secreted in response to thrombin and this release was inhibited by PGE1. The platelet: serum ratio of each Ig and albumin were similar. In grey platelets deficient in α‐granules, PA‐Ig and PA‐albumin levels were raised per platelet but when increased platelet size was taken into account PA‐Ig were normal and PA‐albumin just below normal. Although thrombin caused release of most of the small amounts of β‐thromboglobulin present, PA‐Ig and PA‐albumin were not released. This suggests that PA‐Ig and albumin from plasma may enter a pool of secretory proteins in normal platelets, but in grey platelets they remain in some other site.
Proerythroblast stimulating activity: its purification from mouse serum and its effect on mouse erythroid cell proliferation in vitro
Authors: Udupa K. B. , Lipschitz D. A.
Published: March 2008

Summary. A factor in mouse serum which stimulates proerythroblast proliferation in in vitro culture (proerythro‐blast stimulating activity‐PSA) was purified by green A dye ligand and high performance liquid chromatography (HPLC). As judged by HPLC, PSA obtained after these two steps appeared to be a homogenous protein of Mr 100 000. It increased the proliferation of proerythroblasts when added alone to a liquid culture of bone marrow which was treated with anti‐RBC antibody to remove haemoglobin containing erythroid cells. PSA functioned synergistically with erythro‐poietin (Ep) so that when added to culture together proliferation was 10‐fold higher than when added to culture alone. PSA also increased CFU‐E number but only when added together with Ep. Dose–response studies indicated that PSA increased CFU‐E number when Ep remained constant and vice versa. PSA addition to culture could be delayed by as much as 12 h without any decrease in the number of CFU‐E colonies that developed. When PSA was added to BFU‐E cultures at the time of culture initiation no increase in BFU‐E was seen. If, however, PSA was added 5 d after culture initiation BFU‐E colonies were significantly increased. These results demonstrate that there is a factor (PSA) which can be purified from mouse serum which is not Ep. Its major site of action appears to be the more mature erythroid precursors that have the capacity to divide.
More familial leukaemia in ALL patients with both unexplained high transcortin levels and an HLA antigen Cw3
Authors: De Moor P. , Boogaerts M. , Louwagie A.
Published: March 2008

Summary. In a group of 74 adult ALL patients 25 had a transcortin level ±3 SD above the normal mean, 15 expressed an HLA‐Cw3 antigen and 11 combined both characteristics. In this subgroup of 11 ALL patients, five had familial leukaemia, i.e. at least one relative with acute leukaemia or lymphoma. On the other hand, only three examples of familial leukaemia were found in a series of 59 ALL patients with one or none of these two characteristics (relative risk: 15.6; P<0.001).
Fibrinolysis during normal human pregnancy: complex inter‐relationships between plasma levels of tissue plasminogen activator and inhibitors and the euglobulin clot lysis time
Authors: Wright J. G. , Cooper P. , Astedt B. , Lecander I. , Wilde J. T. , Preston F. E. , Greaves M.
Published: March 2008

Summary. Although it has been previously considered that blood fibrinolytic capacity is reduced during pregnancy, this has been disputed. Also the mechanisms underlying any change in fibrinolysis in pregnancy require clarification. We have therefore measured the plasma activity of tissue plasminogen activator (t‐PA) and inhibitors (t‐PAi) and the concentration of the pregnancy specific inhibitor (PA12) antigen, as well as the euglobulin clot lysis time (ECLT) during normal pregnancy. Plasma concentrations of fibrinogen, plasminogen, fibrin(ogen) degradation products (FDP) and cross‐linked products (D‐dimer) were also monitored. We confirm a marked reduction of the fibrinolytic activity of the plasma euglobulin fraction from the second trimester, and a parallel reduction in t‐PA and increase in t‐PAi activities, with rapid return to non‐pregnant levels post‐partum. In contrast, PAI2, whilst undetectable in non‐pregnant control plasma, was already measurable in the first trimester, increased through pregnancy, and remained at a high concentration up to at least 48 h post‐partum. Fibrinogen and plasminogen concentrations rose progressively through pregnancy and FDP and D‐dimer were frequently detectable in late pregnancy plasma. Changes in the ECLT and plasma t‐PA and t‐PAi activities in pregnancy cannot therefore be directly related to the concentration of PAI2 antigen. Also, despite the apparent marked reduction in fibrinolytic capacity fibrin(ogen) breakdown products are frequently present in increased plasma concentrations in late pregnancy.
Transferrin receptor expression on AML blasts is related to their proliferative potential
Authors: Kozlowski R. , Reilly I. A. G. , Sowter D. , Robins R. A. , Russell N. H.
Published: March 2008

Summary. Expression of the transferrin receptor (TfR) was studied on peripheral blood blast cells from 11 patients with acute myeloid leukaemia (AML). Using a monoclonal anti‐TfR antibody (OKT9) and a polyclonal antibody against surface membrane‐bound transferrin, a proportion of blasts from all the patients was found to express receptors for transferrin. Further analysis of OKT9 expression using a fluorescent activated cell sorter (FACS) showed that the TfR was heterogeneously distributed in the blast cell population. In five out of six samples studied, stimulation of DNA synthesis following short‐term culture induced a several‐fold increase in TfR display as analysed by flow cytometry using OKT9 or FITC‐conjugated transferrin. Blasts from seven patients stained with OKT9 were separated on the FACS into positive and negative or weakly positive fractions. Culture of the TfR negative population in a blast cell colony assay produced no colonies in either of two patients. In a further five patients the colony forming cells were predominately associated with the TfR strongly positive fraction (52 ± 25 colonies/104 cells) rather than the TfR weakly positive fraction (12 ± 11 colonies/104 cells). Analysis of colony size showed that clones derived from the weakly positive fraction were smaller than clones from the TfR strongly positive fraction. These results suggest that TfR display by AML blasts is related to their proliferative capacity and is expressed by the leukaemic stem cell fraction.
Vitamin D metabolism in myeloma
Authors: Lawson‐Matthew Peter , Clayton Jill , Guilland‐Cumming Diane , Yates Ashley , Preston Eric , Greaves Michael , Kanis John A.
Published: March 2008

Summary. We measured serum concentrations of vitamin D metabolites in 27 patients with myeloma, 16 of whom were hypercalcaemic. Serum values were compared with values from normal controls. Serum concentrations of calcitriol (1,25‐dihydroxyvitamin D; 1,25(OH)2D3) were decreased in both hyper‐ and normocalcaemic patients, but more markedly so in hypercalcaemia. These findings were not solely due to impaired renal function nor due to an inadequate supply of 25‐hydroxyvitamin D, and in hypercalcaemic patients were reversed by diphosphonate treatment. We conclude that hypercalcaemia in myeloma is not mediated by abnormalities in vitamin D metabolism. Decreased serum concentrations of 1,25(OH)2D3 may adversely affect both skeletal and tumour metabolism.
Rheological properties of erythrocytes in recombinant human erythropoietin‐administered normal rat
Authors: Maeda Nobuji , Kon Kazunori , Tateishi Norihiko , Suzuki Yoji , Sekiya Misuzu , Taniguchi Takuya , Seike Masahiko , Nakajima Takashi , Shiga Takeshi , Tanaka Kouichi , Shinkura Hirofumi
Published: March 2008

Summary. Recombinant human erythropoietin [rhEPO (180 iu/μg); 1 or 10 μg polypeptide equivalent/kg] was intravenously administered daily to 6‐week‐old normal rats for 1 week. Rheologically, (1) blood viscosity at shear rate of 40–380 s−1 increased in a manner entirely dependent upon the haematocrit, (2) no change in erythrocyte deformability was detected by high shear rheoscopy, and (3) the velocity of rouleau formation in autologous plasma (at 7.5 s−1) decreased. Haematologically, rhEPO administration induced considerable polycythaemia with reticulocytosis in a dosedependent manner, accompanying increased cell volume and decreased intracellular haemoglobin concentration, thus the density distribution of erythrocytes shifted towards low specific gravity. Plasma viscosity and plasma protein composition were unaffected by rhEPO‐administration. In erythrocyte metabolism, no drastic alteration in the level of 2,3–DPG or ATP was detected.
Inhibition of CFU‐GM by prostaglandins in a case of chronic T‐cell lymphocytosis and neutropenia
Authors: Smith J. G. , Smith M. A. , James I. , Blundell E. , Maddison P. J.
Published: March 2008

A 45‐year‐old Caucasian female with seropositive rheumatoid arthritis was found coincidentally to have a circulating lymphocytosis (6.4 × 109/1) and neutropenia (0.1 × 109/1). Initial presentation was with mouth ulceration and recurrent infections. A spleen scan showed no evidence of splenomegaly and serum titres against EBV, CMV and toxoplasmosis were negative. No anti‐neutrophil antibodies were found. Marrow aspiration demonstrated a lymphocytosis of 60% with reduced numbers of granular precursors. Lymphocytes in both blood and bone marrow were CD3, CD8, CD16 and HLA‐DR positive. Lymphocyte conditioned medium (LCM) generated from the patient's blood lymphocytes (without phytohaemagglutinin) was found to inhibit allogeneic colony‐forming unit, granulocyte‐macrophage (CFU‐GM) stem cells in semi‐solid culture compared with control LCM. This inhibitory activity was abrogated by the cytolytic removal of CD8 cells prior to LCM production and was significantly reduced by co‐culture with indomethacin. Culture of the patient's marrow at autostimulatory light density marrow cell concentrations showed poor spontaneous CFU‐GM colony formation until marrow CD8 lymphocytes were removed cytolytically. Prednisolone was used therapeutically (40 mg/d) and resulted in the patient's neutrophil count rising from 0.06 × 109/1 to 1.1 × 109/1 and a fall in the total lymphocyte count to 1.9 × 109/1. Re‐evaluation of the patient's LCM post steroid therapy showed loss of the previous inhibitory effect. The patient's neutrophil count is maintained on oral azathioprine and indomethacin.
Serum levels of soluble interleukin‐2 receptor in hairy cell leukaemia: a reliable marker of neoplastic bulk
Authors: Ambrosetti A. , Semenzato G. , Prior M. , Chilosi M. , Vinante F. , Vincenzi C. , Zanotti R. , Trentin L. , Portuese A. , Menestrina F. , Perona G. , Agostini C. , Todeschini G. , Pizzolo G.
Published: March 2008

In hairy cell leukaemia (HCL), the strong membrane expression of the Tac antigen, corresponding to the p55 chain of the interleukin‐2 receptor (IL‐2R), is associated with the presence in the serum of high levels of a soluble form of the same molecule (sIL‐2R). Previous observations that therapy‐induced clinical and haematologic improvement in HCL is accompanied by a progressive decrease of sIL‐2R suggest a possible correlation between sIL‐2R levels and tumour burden. To verify this hypothesis, we monitored the variation of sIL‐2R values in 13 non‐splenectomized HCL patients admitted for treatment with recombinant interferon alpha‐2. The data were correlated with the estimated weight of bone marrow (BM) and spleen infiltrations, which in these patients almost entirely account for the tumour mass. The regression analysis test showed a direct correlation between sIL‐2R values and both BM neoplastic involvement (r= 0.63) and spleen tumour mass (r= 0.76). In addition, the correlation was further improved (r= 0.86) when sIL‐2R values were correlated with the total neoplastic mass, as calculated by the sum of spleen and BM neoplastic tissue weight. These data indicate that the detection of sIL‐2R in HCL is a reliable non‐invasive marker of tumour burden, which can be regarded as an additional useful tool for monitoring treatment response.
Study of three patients with monoclonal gammopathies and ‘lupus‐like’ anticoagulants
Authors: Bellotti V. , Gamba G. , Merlini G. , Montani N. , Bucciarelli E. , Stoppini M. , Ascari E.
Published: March 2008

In three patients with monoclonal gammopathies: a case of multiple myeloma, a case of monoclonal gammopathy of uncertain significance (MGUS) and a case of monoclonal gammopathy associated with lymphocytic lymphoma, we found the presence of a circulating lupus‐like anticoagulant. Coagulative studies showed that the paraproteins: an IgG3k, an IgG1k and an IgMλ, were responsible for the anticoagulant activity by interacting with the thromboplastin phospholipids. Using isoelectrofocusing we demonstrated that the three monoclonal immunoglobulins had a strong basic charge which may have contributed to determining their interaction with the acidic thromboplastin phospholipids. The binding of various phospholipids to the monoclonal proteins was assessed by the fluorescence quenching method which showed heterogeneous specificity. In order to establish whether the electrical charge is also relevant in cases with polyclonal lupus anticoagulant, the polyclonal immunoglobulins were fractionated according to their charge. The strongest inhibitor activity was found in the most basic immunoglobulins. Monoclonal lupus‐like anticoagulants represent useful tools for investigating the heterogeneous world of polyclonal lupus‐like anticoagulants.
Studies on the sensitivity to complement‐mediated lysis of erythrocytes (Inab phenotype) with a deficiency of DAF (decay accelerating factor)
Authors: Merry Anthony H. , Rawlinson Violet I. , Uchikawa Makoto , Daha Mohamed R. , Sim Robert B.
Published: March 2008

No episodes of clinically significant in vivo haemolysis have been reported in individuals with a novel form of decay accelerating factor (DAF) deficiency (Inab phenotype), nor do functional in vitro assays for complement‐mediated haemolysis show the extreme sensitivity to lysis characteristic of paroxysmal nocturnal haemoglobinuria (PNH) erythrocytes. DAF appears to be totally deficient in the Inab erythrocytes as judged by immunochemical and functional assays. Unlike PNH, the only other described DAF deficiency (where several other phosphatidylinositol (PI)‐linked membrane proteins are also absent), the only protein lacking from Inab erythrocytes appears to be DAF. The Inab phenotype seems to be an inherited specific defect in DAF whereas PNH is an acquired defect in the mechanism of insertion of PI‐linked proteins into cell membranes. These findings support the view that susceptibility of PNH erythrocytes to in vivo and in vitro complement‐mediated haemolysis is not due simply to DAF deficiency but to either the combined lack of several membrane proteins or to deficiency of other regulatory proteins such as the membrane attack complex inhibitor/homologous restriction factor (MIP/HRF). The findings also raise questions as to the role of erythrocyte DAF.
Mechanism of action of α interferon in chronic granulocytic leukaemia: evidence for preferential inhibition of late progenitors
Authors: Galvani D. W. , Cawley J. C.
Published: March 2008

The effect of α interferon (α IFN) on colony forming unit, granulocyte‐macrophage (CFU‐GM) formation by normal bone marrow (BM) as compared with chronic granulocytic leukaemia (CGL) BM and peripheral blood (PB) was tested in semi‐solid assay systems employing either 5637CM or recombinant granulocyte‐macrophage colony stimulating factor (GM‐CSF) to support growth. αIFN (> 125 U/ml) caused consistent inhibition (P=0.02) of day‐7(late progenitor) colonies, but had little or no effect on either day‐7 clusters or day‐14 colonies/clusters. This selective effect on day‐7 colonies was quantitatively similar for both normal and CGL (P < 0.5). Similar results were obtained whether or not the mononuclear preparations were depleted of potential accessory cells, suggesting that the αIFN‐suppression is directly mediated. Morphological examination of colonies and clusters showed that IFN had no effect on cell maturation and that colony inhibition is not, therefore, a consequence of blocked maturation. Since the late‐progenitor compartment is preferentially expanded in CGL, we suggest that our demonstration that αIFN selectively inhibits this compartment is relevant to the clinical effects of the cytokine in the disease.
No preferential use of the VH(V) family in human multiple myeloma
Authors: Clofent G. , Brockly F. , Commes T. , Lefranc M. P. , Bataille R. , Klein B.
Published: March 2008

A recently described immunoglobulin VH family (the VH(V) family) close to the DH and JH genes is preferentially rearranged in immature B‐cell tumours. The question of the emergence of multiple myeloma (MM) from a tumorous pre‐B cell is not yet resolved. To draw a comparison with chronic lymphocytic leukaemia (CLL), we studied the VH(V) rearrangements in 28 MM patients. A rearranged Hind III‐Bam HI fragment of 9.5 kb was detected in only one patient instead of the rearranged fragment of 8.5 kb described in CLL. Rearrangements of a member of the VH(V) family in a 9.5 kb fragment were also observed in two out of 20 lymphoblastoid cell lines obtained from peripheral blood of MM patients. We report here that the VH(V) family is not preferentially involved in this pathology and that the size of the only rearrangement obtained is larger than the 8.5 kb fragment observed in CLL. These results do not favour the hypothesis of a pre‐B cell involvement in MM.
A new case of ‘type II’ inherited protein S deficiency
Authors: Chafa O. , Fischer A. M. , Meriane F. , Chellali F. , Rahal S. , Sternberg C. , Benabadji M.
Published: March 2008

Protein S inherited deficiency is associated with high risk of recurrent venous thrombotic disease (Broekmans et al, 1985a, b). Protein S exists as two forms in plasma, either free and functionally active or complexed with C4b‐binding protein (C4b BP) and inactive (Dahibäck & Stenflo, 1981). We report here the case of a 26‐year‐old woman and her brother, 28 years old, both suffering from recurrent venous thrombosis since the age of 20, diagnosed as severe protein S deficiency according to the following data: free protein S: 2.5–3% by ELISA, undetectable by electroimmunodiffusion (BID); total protein S: 13–16% by ELISA, 21–18% by EID, C4b BP: normal levels. Crossed immunoelectrophoresis using anti‐protein S antibodies revealed only traces of protein S associated with C4b BP and no free protein S. All these assays were performed in the absence of any anticoagulant therapy. Among the investigated relatives, less severe protein S deficiency was observed in three children of the propositus: total protein S levels ranging from 41% to 50% (EID), 40–53% (ELISA); free protein S levels ranging from 16% to 18% (EID), 10–12% (ELISA); normal C4b BP levels. Crossed immunoelectrophoresis revealed traces of free protein S but a significant amount of protein S associated with C4b BP. From these results, we consider, according to Comp's classification (Comp et al, 1986a), that the propositus and her brother are the second case of protein S deficiency type II to be reported in the literature while her children belong to the type I category.
Detection of Rh D‐associated epitopes in human and animal tissues using human monoclonal anti‐D antibodies
Authors: Thorpe Susan J.
Published: March 2008

The distribution of Rh D‐associated epitopes in human and animal tissues has been assessed by immunochemical techniques using six human monoclonal anti‐D antibodies. The IgM antibody MAD‐2 reacted predominantly with endothelial cells lining blood vessels and with animal leucocytes and human leucocytes from both Rh D‐positive and Rh D‐negative donors. Immunoblotting revealed reactivity with a 55 kDa tissue component which was most prominent in homogenates of heart, lung and spleen. In contrast, the IgG antibodies UCHD4 and FOG‐1 stained smooth muscle, and UCHD4 in addition weakly stained cardiac and skeletal muscle and the glomeruli of kidney. Immunoblotting with UCHD4 revealed reactivity with a 27 kDa tissue component which was most prominent in heart homogenates. The results show that the six monoclonal anti‐D antibodies recognize at least four different epitopes and that Rh D‐associated epitopes may occur in non‐erythroid cell types.
Effect of antilymphocyte globulin (ALG) on bone marrow T/non‐T cells from aplastic anaemia patients and normal controls
Authors: Tong Jie , Bacigalupo Andrea , Piaggio Giovanna , Figari Osvaldo , Marmont Alberto
Published: March 2008

The aim of this study was twofold: (a) to test the effect of antilymphocyte globulin (ALG) on bone marrow (BM) T/non‐T cells, and (b) to look for a possible differential response of cells from severe aplastic anaemia (SAA) patients and controls. For this purpose bone marrow T/non‐T cells from normal individuals (n=7) or aplastic patients (SAA, n=13) were kept in liquid culture with or without ALG. Supernatants were then tested for enhancement/suppression on colony forming unit, granulocyte‐macrophage (CFU‐GM) growth (in the he presence of exogenous recombinant granulocyte‐macrophage colony stimulating factor (rGM‐CSF)), or for their ability to support CFU‐GM growth (in the absence of exogenous rGM‐CSF). Supernatants from SAA T cells suppressed CFU‐GM growth of normal bone marrow cells in 5/12 patients (mean expected growth (EG) 71 ± 16%), but not after incubation with ALG (mean 110±29% EG, P=0.03). No inhibition could be obtained with the supernatants from untreated normal T cells. Significant enhancement was seen with ALG treated versus untreated SAA T cells (142±28%EG v. 105±61%EG, P=0.01) and with ALG treated versus untreated SAA non‐T cells (165±26%EG v. 105±23%EG, P=0.01), but not in controls.

Supernatants from SAA and control T/non‐T cells were capable of promoting colony formation in the absence of rGM‐CSF (colony‐stimulating activity (CSA) production): 16±14% for SAA‐T cells and 19±18%EG for non‐T cells 100%= 30 ng rGM‐CSF/ml). The addition of ALG increased CSA production in T cells to 37±23%EG (P=0.04) and in non‐T cells to 40±13% EG (P=0.04). Similar results could be obtained in controls.

In conclusion: (a) ALG interacts in vitro with bone marrow T and non‐T cells from SAA patients, down‐regulating the production of negative lymphokines and enhancing the release of haemopoietins; (b) the latter, but not the former effect, can be shown also with cells from normal controls. The two effects are not mutually exclusive, and are likely to provide maximal benefit in vivo.
In vivo and in vitro inhibitory effect of α‐interferon on megakaryocyte colony growth in essential thrombocythaemia
Authors: Gugliotta Luigi , Bagnara Gian Paolo , Catani Lucia , Gaggioli Licia , Guarini Anna , Zauli Giorgio , Belmonte Monica Mattioli , Lauria Francesco , Macchi Silvia , Tura Sante
Published: March 2008

Summary Megakaryocyte (MK) colony growth of bone marrow mononuclear non‐adherent cells was evaluated in 28 patients with essential thrombocythaemia (ET) and in 26 normal controls. The number of MK‐colony forming units (CFU‐MK per 3 × 105 plated cells) was similar in ET (68 ± 33) and in controls (63 ± 37), independently of bone marrow accessory cells. On the contrary, the size of the MK colonies was significantly (P < 0.01) greater in ET patients. Human recombinant α‐interferon 2a (α‐IFN), administered to 10 patients at a dose of 3 × 106 IU/d s.c. for 11 ± 3 weeks, was capable of inducing a significant (P < 0.01) decrease in the number (from 72 ± 16 to 31 ± 14) and size of bone marrow CFU‐MK, together with a significant reduction of the platelet count (from 1031 ± 325 to 378 ± 75 × 109/1). When added in vitro at time 0 to the culture dishes, α‐IFN inhibited the CFU‐MK growth of both normal and ET bone marrow samples, even at very low concentrations (1 and 10 IU/ml). This study demonstrates that α‐IFN, both in vivo and in vitro, exerts an inhibitory effect on the growth of MK progenitors, which appears to correlate with the clinically documented antiproliferative effect of this cytokine.
Peanut agglutinin shows specificity for bone marrow plasma cells
Authors: Rhodes E. G. H. , Flynn M.
Published: March 2008

Summary A panel of lectins was used to study surface carbohydrate expression on myeloma cells with the aim of finding a possible agent for in vitro bone marrow purging. Peanut agglutinin (PNA, galactose β1,3 n‐acetylgalactosamine‐binding) bound to all plasma cells in 33/34 bone marrow samples from myeloma patients and to all plasma cells in 11 bone marrows from patients with monoclonal gammopathy of undetermined significance and 10 normal bone marrow samples. Bone marrow and peripheral blood monocytes reacted weakly with PNA except in one case of acute monoblastic leukaemia and two of chronic myelomonocytic leukaemia in which monocytes were strongly positive. The only case of plasma cell leukaemia studied was PNA negative. All other bone marrow mononuclear cells were negative for PNA but became positive after sialidase treatment. Peanut agglutinin may have potential as an agent to be used in myeloma for in vitro marrow purging prior to autologous transplantation in combination with high dose chemotherapy.
Pre‐B and ‘common’lymphoblastic leukaemia of childhood compared
Authors: Lilleyman and J. S. , Hinchliffe R. F.
Published: March 2008

Summary Consecutive children suffering from pre‐B lymphoblastic leukaemia (pre‐B ALL) were compared with those who had ‘common’ALL (C‐ALL) to assess any clinical and pathological differences between the two groups.

Over 101 months 27 pre‐B children were seen—an incidence in the population served of around 0.8/100 000 per year. There was some time‐clustering and 12 of the 27 presented in one year (1987). The 51 patients with C‐ALL who presented for comparison were distributed more evenly over the study period. Pre‐B children had more basophilic blast cells with less periodic acid‐Schiff positivity. They had higher presenting white cell counts and serum concentrations of lactic dehydrogenase; two variables which were correlated with each other. There was a trend towards pre‐B children being younger and fewer had hyperdiploid blasts, but these differences were not statistically significant. No difference from ‘common’ALL in response to therapy was apparent for the pre‐B patients at a median follow up time of 17 months—too short a period for any conclusion to be drawn.

Other than immunophenotype, pre‐B ALL has no pathognomonic features, but there are differences from C‐ALL in the distribution of some disease characteristics known to be associated with a worse prognosis.
Transforming growth factor‐β regulates growth as well as collagen and fibronectin synthesis of human marrow fibroblasts
Authors: Kimura Akiro , Katoh Osamu , Hyodo Hideo , Kuramoto Atsushi
Published: March 2008

Summary Three growth factors present in platelets, namely platelet‐derived growth factor (PDGF), transforming growth factor‐β (TGF‐β) and epidermal growth factor (EGF), have been implicated in the pathogenesis of bone marrow fibrosis frequently associated with myeloproliferative disorders. In this study, regulation of the proliferation, as well as collagen and fibronectin synthesis from marrow fibroblasts by TGF‐β was investigated. TGF‐β alone at high plating density stimulated the proliferation of cells at low concentrations, but rather showed inhibition at high concentrations in both MPD patients and control subjects. In the presence of PDGF, which has been confirmed to be a main growth factor for marrow fibroblasts, low concentration of TGF‐β inhibited the proliferation at low cell density, but there was no inhibition at high cell density. The synthesis of both type I and type III procollagen was enhanced by high concentrations of TGF‐β in both MPD patients and control subjects, while PDGF or EGF showed no effect. The fibronectin synthesis was also enhanced by TGF‐β, but not by PDGF or EGF. These results suggest that growth and stromal protein synthesis of fibroblasts causing marrow fibrosis are regulated by TGF‐β as well as PDGF and EGF, when these factors are released or leaked from platelets or megakaryocytes into marrow environment in MPD patients.
A new prognostic system for multiple myeloma based on easily available parameters
Authors: Bladé Joan , Rozman Ciril , Cervantes Francisco , Reverter Joan‐Carles , Montserrat Emili
Published: March 2008

SummaryThe prognostic significance of different presenting features in 180 patients with multiple myeloma (MM) from a single institution was analysed. Out of eight variables isolated from the univariate analysis only two (blood urea and serum albumin), were significant in the multivariate model. Derived from these two simple variables, the relative risk of each patient was calculated, and subsequently two subpopulations of patients could be recognized. The first group included patients with a very active myeloma and a high risk of death soon after diagnosis, their median survival being of only 11–6 months, and the second one comprised patients with low risk of death during the first year and a median survival of 28 months. A hazard function derived from two‐thirds of the patient population (training group) was successfully validated in the remaining subset of patients (test group). Finally, the three major available myeloma staging systems (Durie & Salmon's, Merliniet al's, and the one proposed by the British Medical Research Council) were tested in the present series, and only the latter one showed prognostic validity.
Graft‐versus‐host disease prevention by methotrexate combined with cyclosporin compared to methotrexate alone in patients given marrow grafts for severe aplastic anaemia: long‐term follow‐up of a controlled trial
Authors: Storb Rainer , Deeg H. Joachim , Pepe Margaret , Doney Kris , Appelbaum Frederick , Beatty Patrick , Bensinger William , Buckner C. Dean , Clift Reginald , Hansen John , Hill Roger , Longton Gary , Anasetti Claudio , Martin Paul , Loughran Thomas P. , Sanders Jean , Singer Jack , Stewart Patricia , Sullivan Keith M. , Witherspoon Robert , Thomas E. Donnall
Published: March 2008

Summary Forty‐six patients with aplastic anaemia (median age 23 years) were given cyclophosphamide followed by infusion of marrow from an HLA‐identical family member. To evaluate postgrafting prophylaxis for graft‐versus‐host disease (GVHD), the patients were entered into a randomized prospective trial comparing a combination of methotrexate and cyclosporin (n= 22) to methotrexate alone (n= 24). Methotrexate/cyclosporin significantly reduced the incidence and severity of acute GVHD and improved early survival. This report updates the results of the randomized trial with follow‐up ranging from 3 to more than 6 years. The methotrexate/cyclosporin regimen did not interfere with sustained engraftment, and there were no significant differences in the incidence of early or late graft rejection among the two treatment groups (10%v 4%). The incidence of chronic GVHD was higher among methotrexate/cyclosporin‐treated patients (58%v 36%; P=0·18). Two patients in each treatment group still require treatment for chronic GVHD, while treatment is no longer needed in the other patients. Projected 4‐year survival is 73% in patients given methotrexate/cyclosporin compared to 58% in patients given methotrexate alone (P=0·16). Having achieved a reduction in the incidence of acute GVHD and associated early mortality without impairing engraftment, it is clear that future progress in marrow grafting for aplastic anaemia must come in the area of chronic GVHD.
Amodiaquine‐induced immune agranulocytosis
Authors: Rouveix B. , Coulombel L. , Aymard J. P. , Chau F. , Abel L.
Published: March 2008

This report describes two patients who developed agranulocytosis while receiving prophylactic amodiaquine treatment. The neutrophil counts returned to normal in one after stopping the drug while the other died of sepsis. Amodiaquine‐dependent circulating neutrophil IgG antibodies were demonstrated in both patients using the indirect granulocyte immunofluorescence test. The antineutrophil antibody activity was enhanced with the use of the major amodiaquine metabolite, mono‐desethyl amodiaquine. Additional studies showed the activity of the sera to be nondialysable, heat stable, active against autologous as well as allogenic cells, and absent from the convalescent sera. There was no growth inhibition of allogenic myeloid committed progenitor cells (CFU‐GM) following incubation with the patients’ sera, complement and amodiaquine. These results indicate that agranulocytosis can be mediated by a drug‐dependent antibody which affects mature blood cells.
Characterization of deletions of chromosome 7 short arm occurring as primary karyotypic anomaly in acute myeloid leukaemia
Authors: Mecucci Cristina , Van Orshoven Angeline , Boogaerts Marc , Michaux Jean‐Louis , Van den Berghe Herman
Published: March 2008

Three patients with an acute myeloid leukaemia (AML) showed a deletion of the short arm of chromosome 7 with loss of the deleted material. The 7p — anomaly originated from either a terminal or an interstitial deletion and it represented the only karyotypic aberration in all the three cases. According to the clinical, morphological and immunological features of this series of patients, a 7p — chromosome appears to be associated with a group of AML with myelodysplastic features in the bone marrow, including secondary disorders in patients treated for a previous malignancy.
VAD chemotherapy for refractory multiple myeloma
Authors: Lokhorst H. M. , Meuwissen O. J. A. Th. , Bast E. J. E. G. , Dekker A. W.
Published: March 2008

Thirty‐four patients with refractory multiple myeloma were treated with 4‐d continuous infusions of vincristine and adriamycin in combination with 4‐d pulsed high‐dose dexamethasone (VAD). Of 31 evaluable patients, 16 entered a complete remission (50%) and three a partial remission (10%). No difference in response rate was observed between primary refractory and relapsed patients. The median response duration was 9 months and the median survival of the responding patients was 12 months versus 4 months for the non‐responders. Ten patients have currently survived longer than 360 d, of which six are stable in complete remission without therapy. All responding patients showed a remarkable improvement of their performance status and 70% of these patients became pain‐free. Bacterial infection was the major complication and was probably due to the intensive corticosteroid programme. Severe myelosuppression was rarely observed. Irrespective of the response to VAD, a high β‐microglobulin of 4 μg/ml or more was a bad prognostic parameter. As early relapses were seen especially in this group of patients, in the patients with a plasma‐cell LI% of 3 or more, and in patients with previous anthracyclin treatment, early consolidation, with, for instance, high dose melphalan, might improve the prognosis for these patients.