bluuebunny/crossref_metadata_2025 huggingface dataset:
Screening for iron deficiency: an analysis based on bone‐marrow examinations and serum ferritin determinations in a population sample of women
Authors: Hallberg Leif , Bengtsson Calle , Lapidus Leif , Lindstedt Göran , Lundberg Per‐Arne , Hultén Lena
Published: March 2008

SummaryEfficacy of different methods in screening for iron deficiency was re‐examined in a randomly selected sample of 38‐year‐old women (n= 203) with known iron status based on absence/presence of stainable iron in bone‐marrow smears. The study was made in 1968–69. Serum ferritin (SF) was determined in 1978 in frozen sera using the Kamco IRMA and, in 1992, samples were re‐analysed using a RIA calibrated with the International Standard 80/602 for SF determination. The effect of storage on SF was calculated from a previously established relationship (courtesy of Dr Mark Worwood. Cardiff) between the results obtained with the Ramco assay and assays calibrated with IS 80/602. The distributions in iron replete and iron deficient women showed less overlap (diagnostic efficiency 91%) for SF than for other haematological parameters. The best discrimination was obtained at SF<16 μg/l (specificity 98%: sensitivity 75%). Absence of iron stores was associated with signs of an iron deficient erythropoiesis. starting already at SF 25–40 μg/l. Use of multiple criteria to diagnose iron deficiency falsely reduces prevalence figures for iron deficiency.
Lack of evidence for a role of HCV in hepatitis‐associated aplastic anaemia
Authors: Pol Stanislas , Thiers Valérie , Driss Françoise , Devergie Agnès , Berthelot Pierre , Bréchot Christian , Gluckman Eliane
Published: March 2008

Summary Hepatitis‐associated aplastic anaemia (HAAA) is usually related to non‐A, non‐B hepatitis agents but the prevalence of anti‐hepatitis C virus (HCV) antibodies is similar in HAAA and aplasia of other origins according to the results of the ELISA1 assay. This fact could reflect either that HCV is not involved in HAAA or that HAAA‐associated defective immune function could yield false negative assays despite on‐going HCV infection.

To test these hypotheses, we compared 19 patients with HAAA, including three ELISAl‐positive, to 23 patients with aplasia of known and unknown origin, including eight ELISA1‐positive. who were matched for age, sex and blood transfusion units. HCV infection was searched for by the second‐generation recombinant‐immunoblot assay (RIBA2), and by the polymerase chain reaction which detects HCV viraemia. HCV viraemia was detected in four among the 19 patients with HAAA and in six among the 23 patients with aplasia of other causes.

HCV does not clearly appear responsible for hepatitis‐associated aplastic anaemia which could be due to non‐A. non‐B, non‐C hepatitis virus. HCV viraemia is frequent in severe aplastic anaemia, even without detectable anti‐HCV antibodies, and reflects mainly transfusion‐associated HCV infection.
Treatment of aplastic anaemia (AA) with antilymphocyte globulin (ALG) and methylprednisolone (MPred) with or without androgens: a randomized trial from the EBMT SAA working party
Authors: Bacigalupo A. , Chaple M. , Hows J. , Van Lint M. T. , McCann S. , Milligan D. , Chessells J. , Goldstone A. H. , Ottolander J. , van't Veer E. Th. , Comotti B. , Coser P. , Broccia G. , Bosi A. , Locasciulli A. , Catalano L. , Battista R. , Arcese W. , Carotenuto M. , Marmont A. M. , Smith E. C. Gordon
Published: March 2008

Summary134 patients with acquired aplastic anaemia (AA) were given HALG 15 mg/kg/d for 5 d and methylprednisolone for 1 month, and randomized to receive (n= 69) or not (n= 65) oxymetholone 2 mg/kg/d p.o. daily for 4 months. Early mortality (<120 d) was comparable in the two arms 12/69 (17%) and 11/65 (17%), and correlated with the severity of the disease (39%, 10% and 6% respectively in patients with neutrophil counts (PMN) <0.2, 0.2–0.5, >0.5 × 109/l). The response rate at 120 d was significantly greater in patients receiving androgens (56%v40%;P= 0.04); it was 68%v48% (P= 0.02) in patients surviving 120 d, and 78%v27% (P= 0.03) in females with PMN less than 0.5 × 109/l. In a multivariate Cox analysis on patients with less than 0.5 × 109/l PMN, the probability of responding without androgens was reduced compared to the androgen treatment arm (P= 0.05).

Survival was comparable in the two groups (71%v65%). It was superior (74%v50%), but not significantly (P= 0.1) in females with PMN 0.5 × 109/l receiving androgens.

Side‐effects, including biochemical abnormalities and virilization, could be controlled and were reversible.

In conclusion, the addition of androgens to HALG and methylprednisolone as first line treatment of aplastic anaemia significantly improves the response rate at 4 months, particularly in females with low neutrophil counts, although there is no significant effect on short‐term survival. The reversible side‐effects warrant the use of androgens as an adjunct to the first course of ALG in females with severe AA.
MDS‐macrophage derived inhibitory activity on myelopoiesis of MDS abnormal clones
Authors: Ohmori Seiichi , Ohmori Masami , Yamagishi Morihisa , Okuma Minoru
Published: March 2008

Summary. We studied the effect of myelodysplastic syndrome (MDS)‐derived adherent cells on colony formation of granulocyte‐macrophage progenitors (CFU‐GM) in both normal and MDS bone marrow cells. MDS‐adherent cells suppressed the growth of normal CFU‐GM colony formation. Antibodies against ferritin almost totally neutralized the haematopoietic inhibitory activity. Antibody against gamma‐interferon (γ‐IFN) did not have such effect.

By cytogenetic analysis using G‐staining method, MDS‐derived CFU‐GM colony showed abnormal clones. MDS have been recognized to be a mosaic of normal and abnormal clones. MDS‐macrophages suppressed the growth of progenitor cells derived from normal clones by soluble factors, but did not suppress the growth of those from abnormal clones. It is suggested that progenitor cells derived from abnormal clones are freed from the negative myelopoietic regulator that may be related to the progress of leukaemia.
Up‐regulation of IL‐6‐receptors by IL‐3 on a plasma cell leukaemia cell line which proliferates dependently on both IL‐3 and IL‐6
Authors: Kobayashi Masanobu , Tanaka Junji , Imamura Masahiro , Maeda Shiro , Iwasaki Hiroshi , Tanaka Masanori , Tsudu Yuuzou , Sakurada Keisuke , Miyazaki Tamotsu
Published: March 2008

Summary. In this report we show that IL‐3 up‐regulates the expression of IL‐6‐receptors on a plasma cell leukaemia cell line which proliferates in response to both IL‐3 and IL‐6. Quantitative binding studies and Scatchard analysis revealed that HSM‐2.3 has a single class of high‐affinity IL‐6 binding sites (200/cell, dissociation constant at the equilibrium 1.43 × 10−11 m). After stimulation with IL‐3, HSM‐2.3 has 1200 binding sites. IL‐6 receptor mRNA expression was detected in HSM‐2.3 only after stimulation with IL‐3.
Colony‐stimulating factor enhancement of myeloid effector cell cytotoxicity towards neuroectodermal tumour cells
Authors: Baldwin Gayle Cocita , Chung Grace Y. , Kaslander Caryn , Esmail Tasnim , Reisfeld Ralph A. , Golde David W.
Published: March 2008

Summary. We conducted experiments to determine the optimal conditions for colony‐stimulating factor‐enhanced neutrophil‐ and mononuclear phagocyte‐mediated antibody‐dependent cell‐mediated cytotoxicity (ADCC) using monoclonal antibodies to disialogangliosides expressed on neuroectodermal tumour target cells. Neutrophil ADCC was most effective at effector: target ratios of 100:1, with maximal cytotoxic responses to melanoma target cells generated by 3 h. Granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) and granulocyte colony‐stimulating factor (G‐CSF) were the most potent stimulators of neutrophil ADCC, and enhanced ADCC activity was inhibited in the presence of antibody to Fc receptor type II (FcRII). GM‐CSF and macrophage colony‐stimulating factor (M‐CSF) treatment of freshly isolated monocytes inhibited antibody‐independent cytotoxicity but enhanced antibody‐dependent responses. After 3 d in culture with CSF, 3–10‐fold enhancement of ADCC against melanoma target cells was observed at effector: target cell ratios of 10:1. Greatest stimulation of macrophage ADCC was obtained when GM‐CSF, M‐CSF or interleukin 3 (IL‐3) were used in conjunction with a secondary stimulus. Although gamma interferon (γ‐IFN) did not augment the cytotoxic capability of GM‐CSF‐ and IL‐3‐stimulated macrophages, prominent cytotoxic enhancement was seen when M‐CSF‐stimulated macrophages were exposed to γ‐IFN. A chimaeric mouse/human monoclonal antibody was found to be equivalent in activity to the murine antibody in neutrophil ADCC; however, in macrophage ADCC assays with submaximal effector cell stimulation, the chimaeric antibody was associated with a two‐fold greater response. These studies indicate that under specific conditions, CSFs capable of increasing the number and functional activity of mature myeloid effector cells enhance antibody‐dependent cytotoxicity to neuroectodermal tumour target cells.
The expression of cytokine and cytokine receptor genes in long‐term bone marrow culture in congenital and acquired bone marrow hypoplasias
Authors: Stark R. , Andre C. , Thierry D. , Cherel M. , Galibert F. , Gluckman E.
Published: March 2008

Summary. A number of cytokines have been shown to have stimulatory activity on multipotent haematopoietic precursors. These include kit ligand (KL), interleukins (IL) 1, 3 and 6 and granulocyte macrophage‐colony stimulating factor (GM‐CSF). Using reverse transcriptase/polymerase chain reaction method (RT/PCR) we have examined the expression of these cytokines, the c‐kit and IL‐6 receptors, in long‐term bone marrow culture (LTC) adherent layer cells in human bone marrow hypoplasia syndromes.

Disorders studied include Fanconi's anaemia (FA, n= 16), idiopathic aplastic anaemia (AA, n= 11), Seckel's syndrome (n= 2), dyskeratosis congenita (n= 2), Shwachman‐Diamond syndrome (n= 1), thrombocytopenia with absent radii syndrome (n= 1), acquired amegakaryocytosis (n= 1), paroxysmal nocturnal haemoglobinuria (n= 1) and acquired agranulocytosis (n= 1).

IL‐6 and GM‐CSF expression appeared reduced in most patients with FA, suggesting that impaired production of these cytokines may contribute to the bone marrow failure seen in most patients with FA. In contrast, abundant IL‐6 and GM‐CSF expression were seen in most patients with AA when compared with the FA group and controls; these may be mediators of a stromal response in this disorder. No obvious differences were seen between the different patients’groups and controls in expression of the other cytokines or cytokine receptors studied.
Summarizing data on survival, relapse, and chronic graft‐versus‐host disease after bone marrow transplantation: motivation for and description of new methods
Authors: Pepe Margaret Sullivan , Longton Gary , Pettinger Mary , Mori Motomi , Fisher Lloyd D. , Storb Rainer
Published: March 2008

Summary. Kaplan‐Meier curves are frequently used to summarize survival, relapse, and chronic graft‐versus‐host disease (GVHD) data in the bone marrow transplant setting. Some fundamental problems with the use of Kaplan‐Meier curves for the summary of relapse and GVHD are described.

Alternatively, the use of simple probability and conditional probability curves for summarizing relapse incidence and transplant‐related mortality is illustrated with previously published data from a randomized clinical trial. Prevalence curves provide an appropriate alternative for the summary of a transient condition such as chronic GVHD. When combined with Kaplan‐Meier curves for survival and/or relapsefree survival, these methods provide a more accurate and comprehensive summary of the various marrow transplant outcomes.

These methods of display are also applicable to other settings where more than one type of treatment failure can occur and where follow‐up time may be variable.
Removal of cytomegalovirus DNA from donor blood by filtration
Authors: Smith Karen L. , Cobain Trevor , Dunstan Robert A.
Published: March 2008

Summary. Blood from five donors, previously shown to be positive for cytomegalovirus (CMV) DNA following polymer‐ ase chain reaction (PCR) amplification, was filtered through commercially available leucocyte filters. Analysis of pre‐ and post‐filtration samples by PCR with ethidium bromide staining has shown that filtration was successful in removing CMV DNA from all samples. This is evidence that leucocyte filtration of red cell concentrates may greatly decrease the risk of CMV disease following transfusion to susceptible patients.
Correction of the DNA synthesis defect in vitamin B12 deficiency by tetrahydrofolate: evidence in favour of the methyl‐folate trap hypothesis as the cause of megaloblastic anaemia in vitamin B12 deficiency
Authors: Hoffbrand A. V. , Jackson B. F. A.
Published: March 2008

Summary. The critical disturbance of folate metabolism caused by vitamin B12 deficiency which results in megalo‐ blastic anaemia remains controversial. Vitamin B12 is required in the methionine synthase reaction in which homocysteine is converted to methionine and methyl tetrahydrofolate (methyl THF) to THF. The ‘methyl‐folate trap’hypothesis suggested that failure of demethylation of methyl THF with consequent deficiency of folate co‐enzymes derived from THF is the crucial lesion caused by vitamin B12 deficiency. A more recent theory suggested that reduced supply of methionine leads to reduced availability of ‘activated formate’and hence of formyl THF and it is this defect that results in failure of folate co‐enzyme synthesis. The present results, based on deoxyuridine suppression tests on 103 cases of megaloblastic anaemia, show that THF itself is equally capable of correcting the failure of thymidylate synthesis in vitamin B12 deficiency as in folate deficiency. Although not as effective as formyl THF in correcting the dU blocking test in vitamin B12 deficiency, this is equally so for the correction of the test by THF compared with formyl THF in folate deficiency. The results therefore favour the theory that it is in the supply of THF and not of ‘active formate’or formyl THF that vitamin B12 plays a critical role in folate metabolism.
Expression of human recombination activating genes (RAG‐1 and RAG‐2) in angioimmunoblastic lymphadenopathy and anaplastic large cell lymphoma of T‐type
Authors: Knecht Hans , Joske David J. L. , Bachmann Edith , Bachmann Fedor , Odermatt Bernhard F. , Pallesen Gorm
Published: March 2008

Summary. In order to determine the genotypic maturation status of the proliferating lymphoid cells in angioimmunoblastic lymphadenopathy (AILD) and in anaplastic large cell lymphoma of T‐type (T‐ALC), recombinase activating gene (RAG‐1 and RAG‐2) expression was assessed in six AILD and five T‐ALC cases using a sensitive reverse transcriptase (RT) and competitive (C) polymerase chain reaction (PCR). RAG transcripts were not detectable in nine cases with high proliferating activity, suggesting that in most cases the proliferating cells are derived from mature (rearranged) lymphocytes. However, low levels of RAG transcripts were detected in one AILD and one T‐ALC case and are consistent with either an involvement of immature lymphoid precursors in the proliferating pool or a deregulated T‐cell maturation pathway with persistence of RAG expression. An association between RAG gene expression and poor response to therapy is possible but has to be tested in larger prospective series.
Heterogeneous abnormalities of platelet dense granule ultrastructure in 20 patients with congenital storage pool deficiency
Authors: Weiss Harvey J. , Lages Bruce , Vicic William , Tsung Lancy Y. , White James G.
Published: March 2008

Studies on platelet dense granule structure were carried out in 20 patients with various types of congenital storage pool deficiency (SPD), including 15 with specific deficiencies of dense granules and dense granule substances (δ‐SPD), and five with combined deficiencies of dense and α‐granules (αδ‐SPD). Dense granules were identified by their high affinity for uranyl ions (uranaffin reaction), by their ability to accumulate the fluorescent dye mepacrine, and by their inherent electron opacity on unfixed, unstained whole mount preparations. By all these methods, dense granules were markedly decreased in seven albino patients with the Hermansky‐Pudlak syndrome (HPS) variant of δ‐SPD. These findings suggest that the basic defect in these patients is a specific abnormality in organelle development which prevents the formation of an intact granule structure, a quantitative abnormality which may differ from that in animals with related pigment disorders. In contrast, eight non‐albino patients with δ‐SPD had, on average, only a slightly reduced number of uranaffin‐positive and mepacrine‐positive granules, but a shift in uranaffin‐granule distribution towards those lacking a dense core (‘empty granules’), suggesting a more qualitative type of dense granule defect. These results are consistent with previous evidence suggesting a decreased uptake of ATP across the granule membrane in δ‐SPD. In addition, on whole mounts, these patients’platelets contained substantial numbers of electron dense chains and clusters which contained P and Ca, but with a P/Ca ratio less than that of typical dense granules, and which were retained, along with a larger amount of ATP, after thrombin treatment of the platelets. The various findings in these patients raise the possibility that these structures may represent microvesicles, derived from the Golgi apparatus, which provide a transport mechanism for concentrating adenine nucleotides and calcium in dense granules and which is impaired in some patients with SPD. Additional defects may account for the more extensive granule abnormalities observed in αδ‐SPD.
Demonstration of autoreactivity by a human monoclonal IgG anti‐Rh D antibody
Authors: Thorpe Susan J. , Bailey Stuart W.
Published: March 2008

Human IgG monoclonal antibodies (mabs) against the Rh D antigen have considerable potential for the prophylaxis of haemolytic disease of the newborn. We have carried out in vitro testing for cross‐reactions with tissue components by screening two such mabs against animal tissues and a wide panel of human organs from nearly 50 individuals, most of whom were of known Rh D phenotype. Cryostat sections were studied by indirect immunohistochemical techniques. One of the mabs showed non‐specific, dose‐dependent binding to multiple tissue components whereas the other specifically and consistently reacted strongly with animal smooth muscle and human smooth muscle (vascular, in the walls of hollow viscera, in the respiratory tract) from both Rh D‐positive and ‐negative donors. Immunoprecipitation experiments identified the probable smooth muscle antigen as actin or actin‐associated. However, on the basis of inhibition experiments, and by direct estimation of the association constant, the affinity of this mab for smooth muscle was lower than that for Rh D. These results demonstrate autoreactivity by an IgG anti‐D mab and show differences in the immunochemical characteristics of human anti‐D mabs which may be clinically relevant.
The prognostic significance of auer rods in myelodysplasia
Authors: Seymour J. F. , Estey E. H.
Published: March 2008

Summary. The category of refractory anaemia with excess blasts in transformation (RAEBt) of the French‐American‐British (FAB) classification system comprises a heterogeneous group of patients: those with any combination of 5% or more blood blast cells, more than 20% but no more than 30% marrow blast cells, or the presence of auer rods and 30% or less marrow blast cells. To determine the prognostic significance of auer rods in RAEBt, we classified the 208 patients with RAEBt seen between 1973 and 1992 as (1) those having RAEBt solely on the basis of auer rods (RAEBta, n = 29). (2) those meeting blood or marrow blast criteria for RAEBt and also having auer rods (RAEBtpos. n = 40) or (3) those meeting blood or marrow blast criteria for RAEBt without having auer rods (RAEBtneg, n = 139). The RAEBta group had a higher survival probability than either of the other two groups. Within RAEBta, those patients who, without auer rods, would be considered RAEB by the FAB system (n = 19) had a higher probability of survival than patients with RAEB as conventionally defined. Furthermore, patients with RAEBtpos were more likely to live longer than those with RAEBtneg. The RAEBta, RAEBtpos and RAEBtneg groups were similar with regard to the usual haematologic parameters. However, patients with auer rods were more likely to have a normal karyotype and less likely to have prognostically unfavourable cytogenetic abnormalities. When analysis was performed within cytogenetic groups, the favourable prognostic impact of auer rods was still evident. Similarly, the favourable prognostic significance of auer rods was discernible both among patients who did not receive intensive therapy and those who received induction chemotherapy. The complete remission rate in auer rod positive patients was 77%, compared to 27% in those without auer rods. There were no differences in remission duration. Our results suggest that: (1) patients with auer rods without blood or bone marrow blast criteria for RAEBt should not be grouped with those patients with such criteria, and (2) patients with auer rods and other criteria for RAEBt have a higher complete remission rate following induction therapy of the type frequently reserved for patients with acute myeloid leukaemia.
Genotyping provides a reliable tool for the determination of the platelet antigen system HPA‐1 in Glanzmann's thrombasthenia
Authors: Pober Michael , Kyrle Paul A. , Panzer Simon
Published: March 2008

Summary. Glanzmann's thrombasthenia (GT) is an inherited disorder of platelet function, characterized by quantitative or qualitative defects of the platelet glycoprotein (GP) IIb/IIIa complex. Patients with GT may require repeated transfusions and therefore alloimmunization against platelet antigens could become of particular interest. As GPIIIa contains the most important platelet alloantigen system, HPA‐1, its diminished expression in GT patients may impede serological determination of the HPA‐1 allotype. By immunofluores‐cence consistent results were obtained in only two out of seven patients. The monoclonal antibody‐specific immobilization of platelet antigen test allowed typing of six patients. DNA analysis was feasible in all cases. All seven patients were identified to be homozygous HPA‐la. Thus, provided a normal HPA‐1 DNA region, its analysis can serve as a reliable tool for HPA‐1 typing in GT even if serological methods fail.
A β°‐thalassaernia due to a 1605 bp deletion of the 5‘β‐globin gene region
Authors: Dimovski A. J. , Efremov D. G. , Jankovic L. , Juricic D. Plaseska. D. , Efremov G. D.
Published: March 2008

Summary. We studied a heterozygous β°‐thalassaemia patient from Croatia with an unusually high Hb A2 level of 7.6% and an elevated Hb F level of 5.8%. The same condition was found in his father (Hb A2 8.2%; Hb F 8.5%). Gene mapping and direct sequencing analyses revealed a new deletion of 1605 bp in the 5′β‐globin gene region between positions —984/5 and +620/1. This deletion has not been observed among more than 500 P‐thalassaemia chromosomes from the Balkan countries studied in our laboratory.
Contrasting interspecies efficacy and toxicology of 1, 2 ‐diethy 1–3 ‐hydroxypyridin‐4‐one, CP9 4, relates to differing metabolism of the iron chelating site
Authors: Porter J. B. , Abeysinghe R. D. , Hoyes K. P. , Barra C. , Huehns E. R. , Brooks P. N. , Blackwell M. P. , Araneta M. , Brittenham G. , Singh S. , Dobbin P. , Hider R. C.
Published: March 2008

Summary. In order to define a predictive animal model for the effects of hydroxypyridinone (HPO) iron chelators in humans, we have compared the 28 d oral efficacy and toxicology of the HPO, 1,2‐diethyI‐3–hydroxypyridin‐4–one (CP94) in rats and guinea‐pigs and related the results to the contrasting metabolism of this compound in the two species. CP94 was highly effective at mobilizing liver iron in rats but showed toxicity at higher doses, whereas in the guinea‐pig the compound lacked toxicity but was ineffective at mobilizing liver iron. These differences can be explained by the contrasting metabolism of the drug between the two species. In rats, at the top dose of 300 mg/kg intragastrically, all animals died before the end of the study, with no deaths or weight loss at lower doses. At 100 mg/kg, rat liver non‐haem iron concentrations were reduced by 53% and 44% in females and males respectively (P<0.001). At this dose, adrenal medullary cell vacuolation, increased mammary secretory activity, vacuolation of corpora luteal cells and single cell hepatocyte necrosis were seen. There were no reductions in the white cell count. At 50 mg/kg rat liver non‐haem iron concentrations were decreased by 50% and 34% in females and males respectively (P<0.02). In female rats this was associated with increased mammary secretory activity. In iron‐overloaded rats given 100 mg/kg by gavage for 28 d, liver non‐haem iron concentration was reduced by 39% (P<0.01) and serum ferritin by 71% (P<0.001). Ovarian and mammary changes were not influenced by iron loading. In guinea‐pigs, CP94 was evaluated at 50 mg/kg, 100 mg/kg or 200 mg/kg by oral insufflation for 28 d. No reduction in liver iron was seen and no systematic dose related histological, biochemical or haematological effects were observed. Whereas in guinea‐pigs 99% of urinary recovery following an oral dose of CP94 (100 mg/kg) was as the inactive glucuronide metabolite, in the rat only 23% of the dose was excreted in the urine as the glucuronide with remainder as the free drug or an iron binding metabolite. The lack of both efficacy and toxicity in the guinea‐pig may therefore be explained by the rapid inactivation of CP94 by glucuronidation. This metabolism of CP94 in the guinea‐pig is closer to humans than the rat, suggesting that both the efficacy and toxicity of this compound in humans may also be limited by glucuronidation.
Impaired G‐CSF production at post‐transcriptional level in a patient with chronic idiopathic neutropenia
Authors: Morikawa Keiko , Yamamori Shunji , Oiwa Kanji , Morikawa Shigeru , Oseko Fumimaro , Kubota Toshibumi , Nakamura Teruhisa
Published: March 2008

Summary. Chronic idiopathic neutropenia (CIN) is a disorder characterized by severe neutropenia and a maturational arrest of the neutrophil precursors in the bone marrow. The pathogenesis of this disorder has been obscure. We examined the production of endogenous G‐CSF and the expression of G‐CSFmRNA in a patient with adult type CIN. The G‐CSF production by the patient's mononuclear cells was deficient in spite of the adequate accumulation of G‐CSFmRNA. Our data suggests that the defect in the endogenous G‐CSF production at the post‐transcriptional level is likely to be an aetiological factor in CIN.
Releasability of human hypereosinophilic eosinophils is related to the density of the cells
Authors: Carlson Marie , Öberg Gunnar , Peterson Christer , Venge Per
Published: March 2008

The activity of eosinophils and neutrophils with respect to the release of granule proteins was studied in 11 patients with the hypereosinophilic syndrome (HES). Granulocytes or purified eosinophils were stimulated with serum‐opsonized Sephadex particles (C3b‐induced release), and the released amounts of eosinophil cationic protein (ECP), eosinophils protein‐X (EPX) and myeloperoxidase (MPO) were measured by means of specific radioimmunoassays (RIA). Eosinophils obtained from patients with HES released significantly more ECP (P<0·002) and EPX (P<0·01) after 20 min of incubation than cells from the control group. The cellular content of ECP and EPX in eosinophils obtained from the patients with HES was significantly reduced to 50% and 62%, respectively, of the content of these granule proteins of eosinophils from the control group. In separated eosinophils light‐density eosinophils released more of both ECP and EPX than normal density eosinophils. There was no difference in MPO release between the patients and the control group. We conclude that the eosinophils from patients with HES have an increased propensity to release their granule proteins and the releasability seems to be related to the density of the cells.
A second course of treatment for childhood acute lymphoblastic leukaemia: long‐term follow‐up is needed to assess results
Authors: Chessells Judith M. , Leiper Alison D. , Richards Susan M.
Published: March 2008

We report the results of long‐term follow‐up of 94 children who completed treatment for acute lymphoblastic leukaemia (ALL) between 1974 and 1986 and subsequently experienced a bone marrow relapse before 1992. 91 children received further induction, intensification and CNS directed therapy; 19 proceeded to BMT or ABMT and the remainder were treated on one of three protocols which increased in intensity. The duration of second remission improved significantly with increasing intensity of treatment and bone marrow transplantion was followed by fewer relapses than chemotherapy. Analysis of factors influencing the duration of second remission showed that only length of first remission was of additional significance; the median duration of second remission being only 19 months in children with a first remission of less than 4 years and 62 months in those with longer first remissions. 29 children electively stopped chemotherapy a second time but only 11 of these remain still in second remission with recurrences occurring for up to 7 years from the the time first relapse. Only three of the 24 longterm survivors had no significant late effects of treatment; these were most marked in children who had received a second course of radiotherapy. We conclude that very long follow‐up is necessary to determine whether patients may be successfully re‐treated following late bone marrow relapse and that all such treatment is associated with a high incidence of late effects.
Treatment of primary refractory and relapsed acute lymphoblastic leukaemia in children and adults: the GIMEMA/AIEOP*experience
Authors: Giona Fiorina , Testi Anna Maria , Annino Luciana , Amadori Sergio , Arcese William , Camera Andrea , Di Montezemolo Luca Cordero , Ladogana Saverio , Liso Vincenzo , Meloni Giovanna , Moleti Maria Luisa , Rondelli Roberto , Zanesco Luigi , Pession Andrea , Mandelli Franco
Published: March 2008

One hundred and forty‐seven patients aged <55 years with advanced acute lymphoblastic leukaemia (ALL) were enrolled in an Italian cooperative study (ALL R‐87), This protocol consists of an induction phase with idarubicin (IDA) plus intermediate‐dose cytarabine (IDARA‐C), followed by a consolidation phase and bone marrow transplant (BMT). Complete remission (CR) was achieved in 97/147 patients (66%) with a CR rate of 77% in children versus 51% in adults (P<0·01), 48 responders (50%) underwent BMT.

Probability of event‐free survival (EFS ± SE) was 10·2±3·1% at 56 months. EFS was 14·3±4·51% at 56 months for children versus 3·8±3·41% at 37 months for adults (P<0·0001). Among patients treated in first relapse, EFS was 14·2±7·79% for patients with CR >18 months versus 6·6±3·17% for those with CR <18 months (P<0·0001).

Projected disease‐free survival (DFS ±SE) was 15·4±4·61% at 55 months for all responders and 43·3±14·34% at 52 months for allografted patients. Projected overall probability of survival ±SE for all patients was 18·8±4·13% at 56 months.

This study confirms the efficacy of IDA plus IDARA‐C in poor‐risk ALL patients. A more intensive post‐remission therapy or alternative approach must be designed to improve long‐term results.
Acute myelogenous leukaemia in the elderly: retrospective study of 235 consecutive patients
Authors: Baudard M. , Marie J. P. , Cadiou M. , Viguié F. , Zittoun R.
Published: March 2008

A retrospective analysis was performed on 235 elderly acute myelogenous leukaemia (AML) patients aged 60 years or more, consecutively admitted to a single haematological department during a 10‐year period from 1980 to 1989. 46% of patients received only conventional induction chemotherapy. The rate of inclusion in EORTC cooperative clinical trials was significantly lower than for younger patients despite specific protocols proposed for the elderly since 1983, thus confirming the important selection bias of most published series on elderly AML patients. Compared with treatment results in patients <60 years. complete remission (CR) rate was lower (33·3%v 65·4%, P<0·0001), with a marked drop in patients older than 70, and induction death rate was higher (21·3%v 12·5%, P= 0·04). Intrinsic characteristics of leukaemic cells, especially expression of the MDR1 gene, in vitro growth of the leukaemic clonogenic cells and sensitivity to daunorubicin+cytosine arabinoside, did not differ according to age, except that there was a higher incidence of previous myelodysplastic syndromes and a lower incidence of good prognostic cytogenetics in the elderly patients. Thus, treatment failure in elderly AML patients appears to be mainly due to host‐related factors (especially performance status and age < or ±70 years), and to inadequate treatments. Some elderly patients may have been undertreated because of the planned anthracycline dose reduction, resulting in a higher rate of ‘resistant’AML, i.e. patients surviving the induction period without entering into CR, than in younger patients (45·4%v 22·1%, P<0·0001). 11 patients (4·7%) with untreated or ‘resistant’AML survived more than 1 year, while receiving only supportive care. These slowly progressive AML patients were characterized by a good performance status, and lower circulating blast cells and bone marrow blast counts.
Cytomegalovirus (CMV) antigenaemia for rapid diagnosis and monitoring of CMV‐associated disease after bone marrow transplantation
Authors: Gondo Hisashi , Minematsu Toshio , Harada Mine , Akashi Koichi , Hayashi Shin , Taniguchi Shuichi , Yamasaki Kazuo , Shibuya Tsunefumi , Takamatsu Yasushi , Teshima Takanori , Eto Tetsuya , Nagafuji Koji , Mizuno Shin‐ichi , Hosoda Kenji , Mori Ryoichi , Minamishima Yoichi , Niho Yoshiyuki
Published: March 2008

A technique for the rapid detection of cytomegalovirus (CMV) antigen‐positive blood leucocytes (CMV antigenaemia) was evaluated in 15 marrow transplant patients as a means of diagnosis and for monitoring CMV‐associated disease. CMV antigenaemia was determined by direct immunoperoxidase staining of leucocytes with a peroxidase‐labelled monoclonal antibody, HRP‐C7, which binds an immediate‐early antigen of human CMV.

CMV antigenaemia occurred in 7/15 marrow transplant patients (47%) and was initially detected between 4 and 6 weeks after transplantation. CMV‐associated diseases developed in 3/15 patients (20%). All patients with CMV‐associated disease had a relatively large number of CMV antigen‐positive leucocytes, exceeding 10 per 50000 white blood cells (WBCs). In the remaining 12 patients, CMV antigen‐positive leucocytes were less than 10 per 50000 WBCs or were undetectable. CMV‐associated disease did not develop in these patients during the period of monitoring. CMV antigen‐positive leucocytes were detected more frequently in patients who developed acute graft‐versus‐host disease (GVHD) or haemorrhagic cystitis than in those without such complications. CMV antigens were detectable from 1 to 4 weeks before the onset of CMV‐associated disease which allowed initiation of ganciclovir treatment at an early stage. The degree of CMV antigenaemia paralleled the clinical symptoms and signs, higher degrees of antigenaemia being associated with more significant disease. Thus, the detection of CMV antigen‐positive blood leucocytes is useful for the diagnosis and monitoring of CMV‐associated disease following bone marrow transplantation.
Virus‐associated haemophagocytic syndrome: further evidence for a T‐cell mediated disorder
Authors: Watson Henry G. , Goulden Nicholas J. , Manson Lynn M. , McDermid George , Gray James A. , Parker Alistair C.
Published: March 2008

A 16‐year‐old youth with life‐threatening virus‐associated haemophagocytic syndrome (VAHS) responded remarkably to treatment with cyclosporin A during two periods of active disease, the second of which was due to non‐compliance with treatment. Our clinical observations support the hypothesis that VAHS is cytokine driven as a result of an aberrant T‐cell response to infection.
The Australian type of nondeletional Gγ‐HPFH has a C→G substitution at nucleotide — 114 of the Gγ gene
Authors: Motum P. I. , Deng Z.‐M. , Huong L. , Trent R. J.
Published: March 2008

Nondeletional hereditary persistence of fetal haemoglobin (HPFH) results in the continued production of 2–25% haemoglobin F (Hb F) in the adult who is heterozygous for this mutation. This increase is associated with single‐base mutations in the promoter region of either the Gγ‐ or Aγ‐ globin genes. Affected positions include – 202, – 175, – 161, – 158 and – 114 of the Gγ gene, and – 202, – 198, – 196, – 195, – 175, and – 117 of the Aγ gene. There is now evidence that these mutations produce their effect by changing the binding of certain regulatory proteins. We describe a novel C→G transversion at position – 114 of the Gγ gene which is associated with the phenotype of Gγ‐ HPFH.
Platelet‐derived growth factor expression in accelerated and blastic phase of chronic myelogenous leukaemia with myelofibrosis
Authors: Kimura Akiro , Nakata Yoshio , Hyodo Hideo , Kuramoto Atsushi , Satow Yukio
Published: March 2008

Summary Myelofibrosis is sometimes associated with accelerated and blastic phase of chronic myelogenous leukaemia (CML). In order to investigate the role of platelet derived growth factor (PDGF) in this pathogenesis, expression and production of PDGF was studied in the blast cells from 11 patients. Five patients had myelofibrosis with myeloid blasts, while six patients did not show fibrosis, including three with myeloid blasts and three with lymphoid blasts. PDGF‐A chain transcript was expressed in most of the patients. On the other hand, PDGF‐B chain transcript was detected in all of the five patients with myeloid blasts and with fibrosis, in one of the three patients with myeloid blasts and without fibrosis, and in none of the three lymphoid crisis patients without fibrosis. In the patients with myeloid blasts and with fibrosis, PDGF protein. PDGF‐AB and/or PDGF‐BB, was found to be secreted from blast cells. In addition, the PDGF activity in the culture of myeloid blasts from two patients with fibrosis was also growth stimulatory for human marrow fibroblasts. These results suggest that expression and secretion of PDGF‐AB or PDGF‐BB in blast cells play an important role in the pathogenesis of marrow fibrosis associated with accelerated and blastic phase of CML.
The role of platelet von Willebrand factor in platelet adhesion and thrombus formation: a study of 34 patients with various subtypes of type I von Willebrand disease
Authors: Fressinaud Edith , Federici Augusto B. , Castaman Giancarlo , Rothschild Chantal , Rodeghiero Francesco , Baumgartner Hans R. , Mannucci Pier Mannuccio , Meyer Dominique
Published: March 2008

Summary In order to investigate the respective role of plasma and platelet von Willebrand factor (vWF) in mediating platelet adhesion and thrombus formation, we performed ex vivo perfusion studies with native blood from patients with various subtypes of type I von Willebrand disease (vWD). We studied 34 patients with type I vWD (19 ‘platelet normal’, five ‘platelet low’, two ‘platelet discordant’, eight ‘Vicenza’). Parallel studies were carried out on nine patients with severe vWD (type III). At high shear rate (2600 s‐1) we found that the defect in platelet‐vessel wall interactions in patients having a normal platelet vWF content (‘platelet normal’ and ‘Vicenza’) involved thrombus formation, whereas platelet adhesion was normal. At this high shear rate, platelet adhesion and thrombus volume were significantly decreased in patients with subtypes ‘platelet low’ and ‘platelet discordant’, i.e. when platelet vWF is either low or dysfunctional. These results indicate that platelet vWF may substitute for plasma vWF to promote platelet adhesion. emphasizing the important role of platelet vWF. They also confirm the role of vWF in thrombus formation at high shear rate because an abnormal thrombus volume was observed in all patients. even when platelet adhesion was normal.
Abnormal cAMP‐induced phosphorylation of rap 1, protein in grey platelet syndrome platelets
Authors: Enouf Jocelyne , Corvazier Elisabeth , Papp Béla , Quarck Rozenn , Magnier Clarice , Kovàcs Tünde , Bredoux Raymonde , Lévy‐Tolédano Sylviane , de Gunzburg Jean , Wuytack Frank , Caen Jacques
Published: March 2008

Summary We previously demonstrated abnormal Ca2+ transport by microsomes in platelets from a grey platelet syndrome patient. Here, we investigated the platelet Ca2+ ATPases that mediate this transport, as well as its possible regulation by rap 1 protein. We showed that grey platelet syndrome platelets expressed the same two distinct Ca2+ ATPases as those recently described in normal platelets; the 100 kD SERCA2‐b isoform (Sarco/Endoplasmic Reticulum Ca2+ATPase) and a new 97 kD SERCA isoform. The two Ca2+ ATPases formed similar amounts of transient phosphorylated intermediates. The expression of these two Ca2+ ATPases was compared by Western blotting using specific antibodies, which again emerged in similar amounts in normal and grey platelet syndrome platelets. As regards the protein phosphorylated by cAMP, it was found to be identical to rap 1 protein when it was immunoprecipitated with an antibody raised against a synthetic peptide specific for rap 1 protein. Although the expression of rap 1 protein was similar in membranes isolated from grey platelet syndrome and normal platelets, its exogenous phosphorylation by cAMP was abnormal, with a concentration (10 μg/ml) of the catalytic subunits of the cAMP‐dependent protein kinase (C.Sub.), as it decreased to half the control level.

It is concluded that the abnormal Ca2+ transport found in grey platelet syndrome platelets is not due to the abnormal expression of the Ca2+ ATPases, but is associated with an abnormality of rap 1 protein phosphorylation by cAMP.
A case of POEMS syndrome with thrombocytopenia and biliverdinaemia
Authors: Yasuda Kazuki , Yatomi Yutaka , Matsushima Teruhiko , Yoneyama Akiko , Higashihara Masaaki , Kurokawa Kiyoshi , Nakahara Kazuhiko
Published: March 2008

Summary This report describes a case of POEMS syndrome with extremely rare complications of thrombocytopenia and biliverdinaemia. The thrombocytopenia appeared to be due to immunopathogenesis. Steroid treatment resulted in complete disappearance of the biliverdinaemia in this patient as well as symptoms and signs of POEMS syndrome, suggesting that the biliverdinaemia was related to POEMS syndrome.
FLAG (fludarabine + cytosine arabinoside + G‐CSF) induces complete remission in acute‐phase chronic myeloid leukaemia: a case report
Authors: Visani G. , Tosi P. , Zinzani P. L. , Manfroi S. , Zaccaria A. , Testoni N. , Lemoli R. M. , Rosti G. , Pelliconi S. , Tura S.
Published: March 2008

Summary The adenine nucleoside analogue fludarabine phosphate in combination with cytosine‐arabinoside (Ara‐C) and granulocyte‐colony stimulating factor (G‐CSF) has recently proved effective in the treatment of poor‐prognosis acute non‐lymphoid leukaemia. We used this triple combination in a case of Ph1 + chronic myeloid leukaemia (CML) unresponsive to alpha interferon that had progressed to acute phase after 5 months of treatment with 6‐mercaptopurine plus hydroxyurea. The patient was treated with four courses of fludarabine 30 mg/m2+ Ara‐C 2 g/m2 (days 1–5) and GCSF (from day 0 to polymorphonuclear (PMN) recovery). Bone marrow blasts decreased from 80% to less than 5%, and karyotyping showed a progressive clearance of Ph1+ metaphases (from 100% to 9% after the fourth course). The patient is presently receiving autologous bone marrow transplantation (ABMT). This therapeutic success in a patient for whom conventional treatment would usually be ineffective makes this combination worthy of further studies, in view of its wider use as a preparative regimen to ABMT in CML.
Clinical and haematological improvement induced by etidronate in a patient with idiopathic myelofibrosis and osteosclerosis
Authors: Sivera Piera , Cesano Luigi , Guerrasio Angelo , Camaschella Clara , Mazza Umberto
Published: March 2008

Summary We report a patient with agnogenic myeloid metaplasia associated with debilitating bone pain due to increased bone turnover and osteosclerosis. Treatment with etidronate at a dose of 6 mg/kg per day on alternate months resulted in a complete recovery of bone symptoms and normalization of metabolic parameters of bone turnover; unexpectedly, a sustained haematological improvement was also observed after several months of therapy, suggesting that bone marrow microenvironment improvement was able to restore a nearly normal haemopoiesis. We suggest that diphosphonate therapy may be of value in patients with AMM and increased bone turnover.
Lack of restricted T‐cell receptor β‐chain variable region (Vβ) usage of reactive T‐lymphocytes in Hodgkin's disease
Authors: Meggetto Fabienne , Al Saati Talal , Rubin Bent , Delsol Georges
Published: March 2008

Summary. T‐cell response against tumour‐associated antigens is mediated by the TCR complex. To determine a possibly restricted TCR‐Vβ repertoire in reactive T‐lymphocytes in Hodgkin's disease (HD), 20 cases (of which 10 were EBV‐positive cases) were investigated using 14 monoclonal antibodies (MoAbs) recognizing 11 different TCR‐Vβ region family products and Northern blot analysis with cDNA probes specific for mRNA transcripts of 11 Vβ families that were not detectable by MoAbs. Four Vβ families (Vβ5, Vβ6, Vβ8, Vβ19) were investigated using both immunohistochemistry (IHC) with anti‐Vβ MoAbs and Northern blot analysis. Immunohistochemical and Northern blot findings were correlated with the detection of the Epstein‐Barr virus (EBV) genome in Hodgkin's and Reed‐Sternberg cells (H‐RS).

The non‐neoplastic lymphocytes in HD were predominantly of T‐phenotype (CD3+). Most of these cells were TCR‐αβ+ (βF1+) and only a few T‐cells were reactive for TCR‐δ1 antibody (TCR‐γδ+). In the majority of cases helper/inducer T‐cells (CD4+) outnumbered suppressor/cytotoxic T‐cells (CD8+). Labelling of these samples with the panel of 14 anti‐Vβ MoAbs showed that only a small percentage (0·2–5·5%) of βF1+ lymphocytes were positive with each of these MoAbs. The proportion of these cells was comparable to that seen in normal tissues. Most TCR Vβ+ cells were randomly distributed, but in virtually all cases occasional Vβ+ cells pertaining to the various Vβ families were seen in close contact to H‐RS cells.

Using total RNA extracted from malignant and normal tissues, no visible band was detected with the various Vβ probes. As determined in the present study, the percentage of T‐cells expressing a given Vβ family must be 10% to be detected with Northern blot. Thus, the percentage of Vβ+ cells expressing Vβ families which were explored only with Northern blot were within the same range as those of the 11 different TCR‐Vβ region families assessed with IHC, i.e. 1–10% of lymphoid cells. The results of the present study show that in HD there is no restricted T‐cell Vβ repertoire usage regardless of the detection of EBV. In addition, since the various Vβ families are represented in T‐cell subpopulations forming rosettes around H‐RS cells, we conclude that the T‐cells attracted by H‐RS cells constitute a polyclonal population.