bluuebunny/crossref_metadata_2025 huggingface dataset:
Long‐term bone marrow cultures in aplastic anaemia
Authors: Marsh Judith C.W.
Published: July 2010

Abstract:  The long‐term bone marrow culture (LTBMC) system provides an in vitro physiological model for the study of stromal cell mediated haemopoiesis in patients with aplastic anaemia. The two aspects of haemopoiesis – stromal and stem cell function – can be analysed separately using a modification of LTBMC with cross‐over studies. Patients with aplastic anaemia universally demonstrate defective stem cell function in terms of reduced or absent marrow repopulating ability, reflecting a deficiency of long‐term culture initiating cells. Defects in stromal cell function, as assessed by the ability of aplastic anaemia stroma to support normal generation of haemopoietic progenitors, are not common, but may conceal an isolated deficiency of a particular growth factor in some patients due to the overlapping nature of haemopoietic growth factor activities. The stem cell abnormality in aplastic anaemia reflects a deficiency in cell numbers, as well as dysfunction in certain cases. An increased level of apoptosis in aplastic anaemia marrow CD34+ cells exists, and this correlates well with disease severity. LTBMC studies demonstrate that more of the haemopoietic cells are nonviable (apoptotic and dead) compared with normal controls, and this correlates with reduced colony (CFU‐GM) generation. An increase in apoptosis among primitive haemopoietic cells may contribute to the stem cell defect in aplastic anaemia. Haemopoietic growth factors such as G‐CSF, when given after immunosuppressive therapy such as antilymphocyte globulin and cyclosporin for aplastic anaemia, may act partly by reducing the increased level of apoptosis, resulting in improved stem cell survival.
Reactive metabolites and agranulocytosis
Authors: Uetrecht Jack P.
Published: July 2010

Central to most hypotheses of the mechanism of idiosyncratic drug‐induced blood dyscrasias is the involvement of reactive metabolites. In view of the reactive nature of the majority of such metabolites, it is likely that they are formed by, or in close proximity to the blood cells affected. The major oxidative system of neutrophils generates hypochlorous acid. We have demonstrated that the drugs associated with the highest incidence of agranulocytosis are oxidized to reactive metabolites by hypochlorous acid and/or activated neutrophils. There are many mechanisms by which such reactive metabolites could induce agranulocytosis. In the case of aminopyrine‐induced agranulocytosis, most cases appear to involve drug‐dependent anti‐neutrophil antibodies, and these are likely to be induced by cell membrane antigens modified by the reactive metabolite of aminopyrine. The target of agranulocytosis associated with many other drugs is usually neutrophil precursors and may involve cytotoxicity or a cell‐mediated immune reaction induced by a reactive metabolite. In the case of aplastic anaemia, there is evidence in some cases for involvement of cytotoxic T cells, which could either be induced by metabolites generated by neutrophils, or more likely, by reactive metabolites generated by stem cells.
Anti‐cytochrome P450 autoantibodies in drug‐induced disease
Authors: Beaune P. H. , Lecoeur S. , Bourdi M. , Gauffre A. , Belloc C. , Dansette P. , Mansuy D.
Published: July 2010

Abstract: Drugs may induce hepatitis through immune mechanisms. In this review we have used the examples of 2 drugs to elucidate the first steps leading to the triggering of such disease, namely tienilic acid (TA) and dihydralazine (DH). These drugs are transformed into reactive metabolite(s) by cytochrome P450 (2C9 for TA and 1A2 for DH) (step 1). The reactive metabolites produced are very short‐lived and bind directly to the enzymes which generated them (step 2). A neoantigen is thus formed which triggers an immune response (step 3), characterized by the presence of autoantibodies in the patient's serum (step 4). The autoantibodies are directed against the cytochrome P450 which generated the metabolite(s). Although the process by which TA and DH induce hepatitis has been elucidated, further studies are necessary to generalize this mechanism. In addition, an animal model will also be useful to fully understand the immune mechanism of this type of disease.
Pharmacogenetics and blood dyscrasias
Authors: Spielberg Stephen P.
Published: July 2010

Abstract: Pharmacogenetic differences in the handling of and response to drugs can markedly alter the risk of severe idiosyncratic adverse drug reactions, including neutropenia, agranulocytosis and aplastic anaemia. Inherited deficiencies of drug metabolizing enzymes can shunt the metabolism of drugs to metabolites which are directly toxic (e.g. 6‐mercaptopurine metabolism to 6‐thioguanine nucleotides) or towards electrophilic metabolites which can kill cells and/or lead to a host immune response (e.g. sulphonamide metabolism to hydroxylamine metabolites). Defects in detoxification pathways (e.g. glutathione conjugation) similarly can predispose patients to adverse outcomes. The advent of molecular screening tools to define individual (rather than population) risk may lead to the use of clinical laboratory tests to identify/predict idiosyncratic adverse drug reactions.
Mechanisms, chemical structures and drug metabolism
Authors: Pohl L.R. , Pumford N.R. , Martin J.L.
Published: July 2010

Abstract: From the studies that have been done by many laboratories over the last 2 decades, it is now clear that the toxicities produced by many drugs are due to their reactive metabolites. It is thought that, in many cases, reactive metabolites cause toxicity by binding covalently to tissue proteins. However, until recently it was difficult to identify these protein targets. Due to the development of an immunochemical approach, this problem has been overcome, as is illustrated here by studies that have been conducted on the metabolic basis of the idiosyncratic hepatitis caused by the inhalation anaesthetic halothane. The major problem to solve in the future will be to determine how protein adduct formation leads to toxicity. It is possible that protein adduct formation may alter an important cellular function or may lead to immunopathology, as is thought to occur in the case of halothane hepatitis. If an allergic reaction is suspected, purified protein targets of reactive metabolites can serve as antigens for identifying sensitized individuals. This information can be used to prevent not only an allergic reaction to the drug, but possibly cross‐reactions to other drugs that are structurally related. Another important application of these studies is the design of safer alternative drugs that will not produce structurally similar toxic reactive metabolites.
Overview of benzene‐induced aplastic anaemia
Authors: Smith Martyn T.
Published: July 2010

Abstract: The scientific literature is replete with reports of cases of benzene‐induced toxicity to the haematopoietic system. These mainly involve aplastic anaemia, the first cases of which were reported in 1897. At high levels of benzene exposure (air concentration > 100 p.p.m.), the incidence of aplastic anaemia is approximately 1/100 individuals exposed, but this drops precipitously at lower levels of exposure (10–20 p.p.m.) to around 1/10,000. Factors that affect susceptibility may include high liver cytochrome P450 2E1 activity and low folic acid intake. The mechanism of benzene‐induced aplastic anaemia remains unclear, but is likely to involve: (a) metabolism of benzene in the liver; (b) transport of metabolites to the marrow and their secondary activation to toxic quinones and free radicals by peroxidase enzymes; (c) induction of apoptosis, DNA damage and altered differentiation in early progenitor cells; and (d) depletion of the stem cell pool.
Metabolic basis of benzene toxicity
Authors: Ross David
Published: July 2010

Abstract: Potential metabolic mechanisms underlying the haemopoietic toxicity of benzene include bioactivation of phenolic metabolites of benzene by peroxidases in bone marrow and ring opening reactions to generate muconate derivatives. Peroxidase‐mediated activation of phenolic metabolites of benzene generates reactive quinones which can be detoxified by NAD(P)H:quinone acceptor oxidoreductase (NQO1). The major peroxidase enzyme in bone marrow is myeloperoxidase (MPO) and potential target cells for phenolic metabolites of benzene were characterized in bone marrow stroma on the basis of high MPO:NQO1 ratios. MPO was found to be expressed at the level of myeloid progenitor cells in both murine (lineage negative cells) and human (CD34+cells) systems. This suggests that progenitor cells may be relevant targets of phenolic metabolites of benzene resulting in aberrant haemopoiesis. A polymorphism in NQO1 is also described which leads to a complete lack of NQO1 activity. The toxicological significance of this polymorphism with respect to benzene toxicity is under investigation.
The effects of benzene and other leukaemogenic agents on haematopoietic stem and progenitor cell differentiation
Authors: Irons Richard D. , Stillman Wayne S.
Published: July 2010

Abstract:  A characteristic shared by a diverse group of myelotoxic compounds and leukaemogens is the ability to act synergistically with granulocyte–macrophage colony stimulating factor (GM‐CSF) in increasing clonogenic response. Pretreatment of murine or human bone marrow cells with the benzene metabolite, hydroquinone, but not phenol, catechol or trans, trans‐muconaldehyde, results in a selective enhancement of GM‐CSF but not an interleukin‐3 (IL‐3)‐mediated clonogenic response. Clonal enhancement is preserved and magnified in enriched populations of CD34+ cells (> 95% purity), suggesting an intrinsic effect on haematopoietic progenitor cell (HPC) recruitment rather than a secondary effect involving accessory cytokines. Clonogenic enhancement of murine HPCs is not accompanied by alterations in GM‐CSF receptor expression or ligand affinity and appears to be mediated via a p53‐independent mechanism. These observations suggest that hydroquinone treatment alters recruitment and differentiation in a primitive subpopulation of CD34+ cells and are consistent with a role for altered stem cell differentiation in the development of chemically induced myelodysplasias.
Nucleoside transporters, bcl‐2 and apoptosis in CLL cells exposed to nucleoside analogues in vitro
Authors: Petersen Amarette J. , Brown Ross D. , Gibson John , Pope Belinda , Luo Xiao‐Feng , Schutz Leonard , Wiley James S. , Joshua Douglas E.
Published: July 2010

The purine nucleoside analogues fludarabine (Fl) and chlorodeoxyadenosine (2‐CdA) are considered to be cell cycle specific agents which require DNA synthesis for cytotoxicity. However, their efficacy in the treatment of CLL, an indolent lymphoid malignancy suggests additional mechanisms of action. Like cytosine arabinoside (AraC), Fl and 2‐CdA gain access to the cell via a specific nucleoside transporter (NST) protein. To investigate the mode of action of these drugs in CLL, we used a fluorescent ligand for the NST (5‘‐(SAENTA‐ x 8)‐fluorescein) and 3‐colour flow cytometry to determine NST expression on CD5+/CD19+ B‐cells from the peripheral blood (PB) of patients with CLL. NST levels on these cells was found to be not significantly different from normal control lymphocytes (mean = 485±425) vs. (mean = 553±178). Exposure to varying concentrations (0, 3 μM and 30 μM) of Fl and 2‐CdA, however, resulted in an upregulation of NST (mean = 1552±775 with 30 μM FL; mean = 3392±2197 with 30 μM 2‐CdA) after 48 h. “Large” lymphoid cells (not present in normal PB) were found to express significantly more NST (mean = 2540±2861) and have a higher proliferative capacity than “small” cells (mean = 357±517 NST/cell). Incubation of CLL cells with Fl (n = 6) and 2‐CdA (n = 8) in vitro over 48 h also resulted in an increase in the proportion of cells in S‐phase (0 μM = 0.2±0.1; 30 μM FL = 2.4±2.0; 30 μM 2‐CdA = 3.3±1.3) and a significant increase in morphologically identifiable apoptosis. Apoptosis was confirmed by flow cytometric DNA analysis (0 μM = 13±8%; 30 μM FL = 40±20%; 30 μM 2‐CdA = 48±11%). In situ hybridization using a biotinylated cDNA bcl‐2 probe demonstrated that bcl‐2 mRNA expression was markedly decreased in treated cells after 24 h. These studies have demonstrated that: (1) NST expression on CLL lymphocytes is low; (2) in vitro exposure to the analogues increases both the level of NST expression and the % cells in S‐phase; (3) exposure to the analogues downregulates bcl‐2 expression and increases apoptosis.
Sequestration patterns of transfused rat neutrophilic granulocytes under normal and inflammatory conditions
Authors: Løvås K. , Knudsen E. , Iversen P. O. , Benestad H. B.
Published: July 2010

Abstract: The fate of polymorphonuclear neutrophilic granulocytes (PMN) after their mobilization from the bone marrow of healthy individuals is not clearly understood. It has been suggested that there is a continuous utilization of these cells in widespread, subclinical inflammatory foci, where they are ultimately degraded. The goal of the present experiments was to determine whether an alternative ecotaxis (“homing”) exists, namely sequestration and degradation of PMN by mononuclear phagocytes exposed to the bloodstream in the liver, spleen and bone marrow. Blood PMN were collected from donor rats, labelled with 51Cr, and injected i.v. into 2 syngeneic rats, one of them having an induced sterile peritonitis. After various time intervals up to 18 h, the rats were killed and exsanguinated. As expected, we found cell‐bound radioactivity in the inflamed peritoneal cavities, and also a high amount of radioactivity in liver, spleen, and bone marrow. The bone marrow uptake of PMN appeared to be much lower in the inflammation rats than in the normal controls. These findings were confirmed in PMN transfer experiments using PVG rats congenic for the RT7 alloantigenic system. Here, transfused blood leukocytes were traced with fluorescent, monoclonal HIS41 antibodies and flow cytometry. A possible corticosteroid effect on the bone marrow sequestration could not be substantiated. Uptake and degradation of PMN takes place in organs containing phagocytes exposed to the bloodstream. Sequestration of PMN in the bone marrow is apparently down‐regulated in inflammatory states, perhaps increasing the PMN availability to inflamed tissue.
Prevalence of antibodies to cardiolipin in chronic ITP and reactivity with platelet membranes
Authors: Årfors L. , Winiarski J. , Lefvert A. K.
Published: July 2010

Anti‐cardiolipin antibodies (ACLA) have been suggested to play a role in the pathogenesis of thrombocytopenia. When sera from 40 patients with chronic idiopathic thrombocytopenic purpura (ITP) were analyzed for ACLA, these autoantibodies were present in 12 (30%). In 4 sera the antibody activity was restricted to the IgG or IgM isotype, respectively, while 4 of the samples contained both IgG and IgM antibodies. To elucidate the interaction between platelets and ACLA, we studied the reactivity of sera from non‐ITP patients with ACLA, with fragmented platelet membranes. None of them had a concurrent platelet deficiency, but sera from 8 (67%) of them showed increased IgG‐binding to platelet membranes. Absorbtion with cardiolipin reduced membrane binding in 6 (50%). C3 levels were normal, while low C4 values occurred in both groups with significantly lower levels in ACLA‐positive patients (p<0.05). Circulating immune complexes (CIC) were common in both groups.

Conclusion: The prevalence of ACLA is increased in ITP and sera from non‐ITP patients with ACLA react with fragmented platelet membranes. This reactivity is often decreased by absorption with cardiolipin, suggesting that ACLA bind to phospholipid epitopes on platelet membranes.
Infectious complications after 2‐chlorodeoxyadenosine therapy
Authors: Van Den Neste E. , Delannoy A. , Vandercam B. , Bosyl A. , Ferrant A. , Mineur P. , Montfort L. , Martiat P. , Straetmans N. , Filleul B. , Michaux J. L.
Published: July 2010

Abstract: Infection is a common adverse event after therapy with nucleoside analogs, including 2‐chlorodeoxyadenosine (CdA). However, the incidence of CdA‐related infections has been poorly documented. In this study we compare, in the same patient population, the incidence of infectious episodes during the 6‐month period before CdA to their incidence during the 6 months after initiating therapy. Ninety‐five patients with hematological malignancies were studied. The incidence of infectious episodes almost doubled after CdA (0.87 vs. 0.47 during the pre‐CdA period). The following factors were associated with an increased risk of infection after therapy: a history of previous chemotherapy, infection during the pre‐CdA period and a diagnosis of chronic lymphocytic leukemia or of non‐Hodgkin's lymphoma. Age, neutrophil and lymphocyte count at onset of CdA and time interval between diagnosis and therapy with CdA did not correlate with the infectious risk. The pattern of infections was modified after therapy with an increase of herpes virus infections (1 vs. 8 episodes, p = 0.04) and of fever of unknown origin (6 vs. 17 episodes, p = 0.03). In conclusion, a population at high risk for developing infectious complications after CdA therapy can be identified. Specific measures aimed at reducing the incidence of infectious events should concentrate on this population.
Characterization and regulation of interleukin‐4 receptor in adult T‐cell leukemia cells
Authors: Mori Naoki , Shirakawa Fumihiko , Murakami Shuichi , Oda Susumu , Eto Sumiya
Published: July 2010

We studied the expression of the receptor of interleukin (IL)‐4, one of the T cell growth factors, on fresh peripheral blood leukemic cells from adult T‐cell leukemia (ATL) patients. Flow cytofluorometric analysis with a monoclonal antibody to the IL‐4 receptor (IL‐4R) were used to investigate whether expression of IL‐4R on ATL cells is different from that on normal lymphocytes and other types of leukemic cells. Leukemic cells from acute type ATL patients synthesize IL‐4R without stimulation, at levels much higher than normal resting lymphocytes and other types of leukemic cells. Furthermore, leukemic cells from acute type ATL showed higher IL‐4R expression than that of chronic type ATL or human T‐cell leukemia virus type I carriers. In addition, there was correlation between expression of IL‐4R on the cell surface and the proliferative response to IL‐4. Both IL‐4 and IL‐2 induced upregulation of IL‐4R on activated normal T cells but not on ATL cells. These results suggest that abnormal expression of IL‐4R may display different biological activities in ATL compared with other types of leukemia. Furthermore, the high expression of IL‐4R in ATL may be involved in the proliferation of leukemic cells and the leukemogenesis in this disease.
Platelet‐specific antibodies in transfused patients
Authors: Uhrynowska M. , żupańska B.
Published: July 2010

Abstract: Sera of 104 patients with haematological diseases, transfused with platelets and sometimes also with red cells, were prospectively investigated in: the lymphocytotoxicity test (LCT), the platelet suspension immunofluorescence test (PSIFT) and the monoclonal antibody immunobilization of platelet antigens (MAIPA) technique. All tests on admission were negative. The overall incidence of alloantibodies after transfusions was 30.8%. Platelet‐specific antibodies were, however, detected much more rarely (9.6%) than anti‐HLA (21.3%). The specificity against HPA antigens was defined in 4 (3.9%) patients: anti‐HPA‐1a, 2b, 3a and 5b. In the remaining 6 (5.7%) cases only the glycoproteins on platelets were identified (GPIb and GPIIIa) which gave positive reaction with the antibodies. Our observations indicate a rather low incidence of platelet‐specific antibodies after transfusions.
Incidence of endocrine complications and clinical disease severity related to genotype analysis and iron overload in patients with β‐thalassaemia
Authors: Jensen C. E. , Tuck S. M. , Old J. , Morris R. W. , Yardumian A. , De Sanctis V. , Hoffbrand A. V. , Wonke B.
Published: July 2010

Abstract: The incidence of endocrine dysfunction in relation to the detailed genotype of β‐thalassaemia is investigated in this study. In addition, the association of genotype to specific clinical features of β‐thalassaemia is examined, together with the relationship between serum ferritin levels and endocrine complications. Ninety‐seven patients were included, all with transfusion dependent β‐thalassaemia. Patients were divided into 2 categories; group 1 consisted of patients with a β0/β0 genotype with or without a concomitant α‐globin gene deletion as well as patients with β0/β+ or β+/β+ genotype and normal α‐globin chain synthesis. Group 2 included patients with β+/β+ or β+/β0 genotype and one α‐globin chain deletion and those with a moderate amount of β‐globin chain synthesis (β++) and normal α‐globin chain synthesis. The results showed that group 1 patients were more likely to have severe clinical disease (p = 0.005). Sixty‐four patients (66%) had at least 1 endocrine disorder and 39 (40%) had multiple endocrinopathies; the most common abnormality was hypogonadotrophic hypogonadism (HH). There was a significant association between patients with group 1 genotypes and the presence of HH and impaired glucose tolerance or diabetes. A positive correlation was demonstrated between serum ferritin concentrations and the presence of thyroid or parathyroid dysfunction.
Results of a fludarabine induction and α‐interferon maintenance protocol in pretreated patients with chronic lymphocytic leukemia and low‐grade non‐Hodgkin's lymphoma
Authors: Zinzani P. L. , Bendandi M. , Magagnoli M. , Rondelli D. , de Vivo A. , Benni M. , Zamagni E. , Cavo M. , Tura S.
Published: July 2010

Abstract: The activity of fludarabine monophosphate (FLU) and α‐interferon (α‐IFN) in low‐grade non‐Hodgkin's lymphoma (LG‐NHL) and B‐cell chronic lymphocytic leukemia (B‐CLL) has been demonstrated in several clinical trials. In a study of 137 previously treated patients, of whom 77 had B‐CLL and 60 with LG‐NHL, we used FLU as salvage chemotherapy. Dosages of 25 mg/m2 were given in 30‐min infusions for 5 consecutive d. Treatment was repeated every 28 d depending on the patient's clinical status for a maximum of 6 cycles. Entrance to the α‐IFN maintenance portion of the study depended on patient response to initial FLU. All patients who had obtained a complete or partial response after the FLU therapy were randomized to receive α‐IFN or no further therapy. The α‐IFN dose was 3×106 units 3 times per wk until disease progression. At 4 yr with a median follow‐up of 22 months the percentage of patients with persistent response ranged between 20% and 30% among all the responders. Thirty‐five (45%) B‐CLL patients achieved major responses (complete/partial response), as did 29 (48%) of those with LG‐NHL. Among the 64 patients who achieved a good response to initial therapy and who have entered the second part of the trial, there has been a rate of prolongation of remission in favour of maintenance α‐IFN (p = 0.02). FLU therapy is an effective drug inducing remission in pretreated B‐CLL and LH‐NHL patients. However, as with other therapeutic modalities, remission is rarely maintained beyond 2 yr. So far, maintenance α‐IFN has not been shown to produce significantly longer remission after treatment with FLU in LG‐NHL, and there is no trend towards prolonged remission in B‐CLL patients. The role of FLU needs to be further evaluated in the management of lymphoproliferative disorders by introducing it in combination with other drugs (α‐IFN) in the induction phase and in maintenance treatment.
A low blood lymphocyte count is associated with an expansion of activated cytotoxic lymphocytes in patients with B‐cell chronic lymphocytic leukaemia
Authors: Martín Alfredo Prieto , Martín Eduardo Reyes , Arriagada Flavio Carrión , Hernandez‐Fuentes Maria del Puerto , Vilaplana Joan Cháfer , García‐Suárez Julio , Soto Melchor Alvarez‐Mon
Published: July 2010

Abstract: In order to determine the relationships between CD2+ lymphocyte subpopulations and tumour mass, the immunophenotype of natural killer (NK) cells and T lymphocyte subsets was studied in 56 B‐chronic lymphocytic leukaemia (B‐CLL) patients and 38 healthy subjects. The patients were classified according to their blood lymphocyte count (BLC). Forty patients had BLC<30×109/l (low BLC, less tumour mass) and 16 patients had BLC>30×109/l (high BLC, larger tumour mass). The percentage of CD3– CD56+ cells, as well as of CD8+, CD8+CD45RO+ and CD3+CD57+ T subsets in low BLC patients, were higher than those found in high BLC patients. Conversely, the percentages of CD3+HLA DR+, CD4+ and CD4+CD45RO+ lymphocytes were higher in high BLC patients than in low BLC patients. The CD4/CD8 ratio was decreased in low BLC patients while it was increased in high BLC patients and a significant positive correlation was found between their CD4/CD8 ratio and their BLC. We conclude that in low BLC B‐CLL patients there is a decreased percentage of activated helper lymphocytes and an increased percentage of NK cells and activated cytotoxic T lymphocytes. These results suggest a role for NK cells, and helper and cytotoxic T lymphocytes in the control of tumour burden in B‐CLL patients.
Maintenance therapy with alpha‐interferon following first‐line VAD in multiple myeloma
Authors: Kars Ayşe , Çelik İsmail , Kansu Emin , Tekuzman Gülten , Özişik Yavuz , Güler Nilüfer , Barişta İbrahim , Güllü İbrahim , Yalçin Şuayib , Altundaǧ Kadri , Zengin Nurullah , Türker Alev , Hayran Mutlu , Baltali Eşmen , Firat Dinçer
Published: July 2010

Abstract:The aim of this study was to evaluate the response characteristics of vincristine, adriamycin and dexamethasone (VAD) as a first‐line chemotherapy and to determine the efficacy of maintenance alpha‐interferon (α‐IFN) in multiple myeloma (MM). Between January 1985 and December 1994, a prospective trial was performed in stage II and III MM patients. The study population received only VAD with no maintenance therapy before 1990 (n= 31), and those recruited after 1990 (n= 33) were planned to be maintained with α‐IFN (5 mU, 3 times per wk) during the plateau to a maximum of 2 yr. Median follow‐up duration (44 vs. 39 months), time to response (3.4 vs. 3.5 months) and rate of objective response (61.3%, 19/31 and 63.6%, 21/33) were similar in VAD‐only and VAD+IFN groups, respectively. The survival analyses revealed higher median progression‐free (39.6 vs. 12 months) and overall survival (65+ vs. 24 months) durations in VAD+IFN group compared to VAD‐only group. VAD regimen was well tolerated and IFN‐related side effects were reversible. These findings denote that IFN maintenance prolongs the duration of response obtained by VAD.
Acquired haemophilia A in women postpartum: management of bleeding episodes and natural history of the factor VIII inhibitor
Authors: Michiels J. J. , Hamulyak K. , Nieuwenhuis H. K. , Novakova I. , van Vliet H. H. D. M.
Published: July 2010

Abstract: The present study reports on the treatment of bleeding episodes and the natural history of factor VIII inhibitors in 4 patients with acquired haemophilia A postpartum. Low titre type II factor VIII inhibitors in 3 patients and high titre type I inhibitor in 1 patient became apparent immediately to 7 months after delivery. High dose human factor VIII concentrate substitution was effective in controlling bleeding episodes in two cases of factor VIII inhibitor type II, but ineffective in 1 patient with high titre type I factor VIII inhibitor. High dose gammaglobulin intravenously in 1 patient with type II factor VIII inhibitor induced a partial correction of factor VIIIc levels for 2 wk. Immunosuppressive treatment in all 4 patients with acquired haemophilia A postpartum did not reduce the potency of the factor VIII inhibitors. The low titre type II inhibitors spontaneously disappeared in all 3 patients within a few months to 1 yr after discontinuation of the immunosuppressive treatment. The high titre type I factor VIII inhibitor persisted for more than 24 yr. We conclude that immunosuppression in 4 women with acquired haemophilia A postpartum did not significantly affect the factor VIII inhibitor titre.
Survival in 86 patients, aged 15–60, with primary acute myelogenous leukemia, treated with a common program in the Norwegian health regions I, III, IV and V in the period 1990–1995
Authors: Tangen J. M. , Wisløff F. , Abdelnoor M. , Brinch L. , Dahl I. M. , Lamvik J. , Ly B. , Nesthus I.
Published: July 2010

Abstract: Eighty‐six patients between 15 and 60 yr with primary acute myelogenous leukemia in health regions I, III, IV and V in Norway were treated according to a common protocol from 21 January 1990 until 1 September 1995 (until 1 January 1993 for health region IV). Seventy‐one percent of the patients reached complete remission (CR) and went on to receive consolidation treatment. In addition to chemotherapy, 18 patients under the age of 52, i.e. 28% of all patients in this age group, received allogeneic bone marrow transplantation. A follow‐up analysis was performed by 1 September 1996. The median overall survival was 15 months, estimated 3‐yr survival 30% and estimated survival at 5 yr was 26%. The median duration of 1st CR was 19 months, and the leukemia‐free survival at 3 yr was 29%. At follow‐up 26/86 patients were alive, 18 in 1st CR (4 after BMT) and 8 in 2nd CR (5 after BMT in 2nd, 1 after BMT in early 1st relapse). These results are comparable to many previously published studies, but may be inferior to the results obtained with more intensive consolidation treatment, including high dose Ara C.
Prognostic value of increased soluble thrombomodulin and increased soluble E‐selectin in ischaemic heart disease
Authors: Blann Andrew D. , Amiral Jean , McCollum Charles N.
Published: July 2010

Abstract:Endothelial cell dysfunction is likely to be important in the pathophysiology of ischaemic heart disease and increased levels of endothelial cell markers soluble E‐selectin and soluble thrombomodulin may reflect this damage. To determine whether increased levels of these markers were predictive of disease progression, we obtained plasma from 54 patients who had survived a myocardial infarction. Soluble E‐selectin and soluble thombomodulin were measured by ELISA. After 49 months, 24 patients had suffered an additional cardiovascular event such as a second myocardial infarction or requirement for arterial surgery. Soluble E‐selectin was 60±30 ng/mL in patients who suffered an end‐point and was 54±23 ng/mL in those without an end‐point (p= 0.43). Soluble thrombomodulin was 65±24 ng/mL in patients who suffered an end‐point and was 49±19 ng/mL in patients who were free of an end‐point (p= 0.009). The major risk factors for atherosclerosis (hypercholesterolaemia, hypertension, smoking) or peak creatinine kinase levels were unable to predict the development of an end‐point. Using life tables, soluble thrombomodulin had a significant effect on survival free of an end‐point (p= 0.011). We conclude that the measurement of soluble E‐selectin is of limited value in epidemiological studies, and that raised soluble thrombomodulin is a new marker for the progression of atherosclerosis in patients with ischaemic heart disease.
Systemic pneumococcal disease after staging splenectomy for Hodgkin's disease 1969–1980 without pneumococcal vaccine protection: a follow‐up study 1994
Authors: Abrahamsen A. Foss , Høiby E. A. , Hannisdal E. , Jørgensen O. G. , Holte H. , Hasseltvedt V. , Høst H.
Published: July 2010

We surveyed, during 1994, all 325 patients who underwent staging laparotomy with splenectomy for Hodgkin's disease in Norway 1969–80, before pneumococcal vaccine was available in this country. The patients were thus not immunized preoperatively. Of 162 patients (49.8%) who died before 1994, 8 (2.4% of the total study) died from pneumococcal septicaemia and 16 (6.2%) from infections totally. Of 163 patients (50.2%) who were alive in 1994, 158 cooperated and filled in a questionnaire: 22 had been hospitalized for serious infections; 2 with pneumococcal septicaemia, and 6 with pneumonia, although lacking a specified microbiological diagnosis. We observed 325 patients representing 4420 patient‐years, 3066 patient‐years among survivors and 1354 patient‐years among the dead. This resulted in an incidence rate of systemic pneumococcal disease of 226 per 100,000 patient‐years, which is a relative risk of 20.5 compared to the general Norwegian population during 1994. Septicaemia for these patients most often had an abrupt clinical start even for relapse‐free individuals and occurred from 2 to 17 yr after splenectomy (mean 10 yr). The risk of developing an overwhelming pneumococcal septicaemia with high case‐fatality in asplenic patients seems to persist for these patients at about the same level even 15–20 yr after splenectomy. Only 12.7% of the survivors had been given pneumococcal vaccine in the autumn of 1993. Despite the fact that medical journals and media in Norway focused upon the problem of pneumococcal disease in asplenic individuals in the autumn of 1993 and spring of 1994, a substantial proportion of these patients (55.3%) still remained unimmunized when interviewed in the autumn of 1994. None of our systemic pneumococcal disease patients was vaccinated. Our data underline the need for prophylactic immunization with effective vaccines against pneumococcal infection in splenectomized Hodgkin's disease patients.
An in‐frame triplet deletion within the gp91‐phox gene in an adult X‐linked chronic granulomatous disease patient with residual NADPH–oxidase activity
Authors: Jendrossek V. , Ritzel A. , Neubauer B. , Heyden S. , Gahr M.
Published: July 2010

Abstract: In an adult patient suffering from X‐linked chronic granulomatous disease (X‐CGD) with residual activity of the NADPH–oxidase we found an unusual biochemical constellation with a defective gp91‐phox gene. As shown by Western blot using a specific antibody the gp91‐phox protein was normal in PMN. However, NADPH–oxidase activity was reduced and no heme spectrum was detectable. By Southern blot and RFLP analysis of genomic DNA a larger defect within the gp91‐phox gene was excluded. Sequencing of the gp91‐phox cDNA revealed an in‐frame deletion of a TTC triplet in exon VI of the gp91‐phox gene. This mutation indicates the loss of one amino acid (phenylalanine 215 or 216) in the gp91‐phox protein. Sequencing of genomic DNA from the heterozygous daughter of the propositus confirmed this mutation. The absence of a functional cytochrome b558‐spectrum in granulocytes of the patient suggests an involvement of the phenylalanine 216 area in heme binding by gp91‐phox. This is the first mutation described in a X‐CGD patient with absence of a functional cytochrome b558‐spectrum but with detectable gp91‐phox protein and residual NADPH‐oxidase activity.
Relation of haemolytic anaemia and erythrocyte‐bound IgG in α‐ and β‐thalassaemic syndromes
Authors: Chinprasertsuk S. , Wanachiwanawin W. , Pattanapanyasat K. , Tatsumi N. , Piankijagum A.
Published: July 2010

Abstract: There has been evidence that IgG‐mediated phagocytosis plays some part in destruction of erythroid cells and subsequent anaemia in thalassaemia. In this study using direct immunofluorescence and flow cytometry, erythrocyte‐bound IgG was studied in 53 and 33 β‐thal/Hb E and Hb H patients, respectively. The mean percentages of IgG‐positive erythrocytes in both nonsplenectomized (4.4%) and splenectomized (24%) β‐thal/Hb E patients were significantly higher than that in normal subjects (1.0%); p < 0.05 and < 0.001, respectively. Splenectomized β‐thal/Hb E patients had significantly higher percentages of IgG‐positive erythrocytes than the nonsplenectomized patients (p < 0.001). The mean percentages of IgG‐positive erythrocytes in Hb H patients were comparable to those from normal controls. However, patients with α‐thal 1/Hb CS had significantly higher IgG‐positive erythrocytes than those with α‐thal 1/α‐thal 2 (p = 0.008). Splenectomized Hb H patients also had higher IgG‐positive erythrocytes than nonsplenectomized Hb H patients (p < 0.039). During haemolytic crisis in some Hb H patients, high percentages of IgG‐positive erythrocytes were also demonstrated. This study demonstrates the presence of increased IgG on erythrocytes in patients with β‐thal/Hb E and in those with Hb H disease following splenectomy or in haemolytic crisis.
Enhanced hematopoiesis in sublethally irradiated mice treated with various quinolones
Authors: Shalit I. , Kletter Y. , Weiss K. , Gruss T. , Fabian I.
Published: July 2010

Abstract: We compared the effect of 6 quinolones on growth of murine bone marrow (BM) progenitor cells in vitro, and their in vivo effect on repopulation of BM and on survival of sublethally irradiated mice. The addition of clinically attainable concentrations of ciprofloxacin, sparfloxacin or clinafloxacin, in concert with pokeweed mitogen (PWM) to murine spleen cells, resulted in a significant enhancement in colony stimulating activity. A 1.5–1.8 fold increase in the number of myeloid progenitors (CFU–C) was observed in the presence of quinolone–PWM spleen conditioned medium (SCM) (prepared with the above‐mentioned quinolones) compared with control cultures exposed to PWM–SCM only. Three other quinolones showed either no stimulatory effect (fleroxacin, norfloxacin) or had an inhibitory effect (ofloxacin) on CFU–C growth. The stimulatory quinolones share in common a cyclopropyl moiety at position N1 of the quinolone ring. This moiety is lacking in the other 3 quinolones. The secretion of interleukin‐3 (IL‐3) and granulocyte‐macrophage colony‐stimulating factor (GM–CSF) by murine spleen cells exposed to quinolone–PWM–SCM was significantly enhanced with all 6 quinolones. However, this effect was associated with a parallel increase in CFU–C only with ciprofloxacin (10 μg/mL), sparfloxacin (1 μg/mL) and clinafloxacin (0.05 μg/mL). The in vivo activity was assessed in sublethally irradiated mice (650 rad) treated with quinolones for 5 d. The number of CFU–C in BM and the number of peripheral white blood cells (WBC) 8 d post‐irradiation was significantly enhanced in mice treated with ciprofloxacin (45 mg/kg/d), sparfloxacin (22.5 mg/kg/d) and clinafloxacin (11.25 mg/kg/d) compared to saline treated animals (p<0.05). Clinafloxacin at higher dosage (45 mg/kg/d) resulted in a decrease in myeloid progenitors in BM. A similar increase in progenitors and WBC was observed in animals treated with high doses, above clinical relevance, of ofloxacin, and norfloxacin (90 mg/kg/d), and with fleroxacin (45 and 90 mg/kg/d). Quinolone‐treated animals, at the above‐cited doses, showed enhanced survival on d18 compared to saline treated animals. The only exception was the higher mortality of clinafloxacin‐treated mice. The above observations imply that certain quinolones, sharing specific molecular structure, are potential immunomodulators at clinically relevant concentrations. These compounds should be further studied in neutropenic patients and BM or peripheral blood progenitor cell recipients.
Small‐dose oral iron absorption test in anaemic and non‐anaemic elderly hospitalized patients
Authors: Joosten E. , Van der Elst B. , Billen J.
Published: July 2010

Abstract:The small‐dose (20 mg) oral iron absorption test (OIAT) was performed in 76 hospitalized elderly patients and 30 healthy adults. Of the elderly patients, 34 were considered as iron deficient (serum ferritin level <20 μg/L) of whom 23 were anaemic and 11 not anaemic, 21 had the anaemia of chronic disorders (ACD) and another 21 were non‐anaemic patients with a normal serum ferritin level. There was a significant inverse correlation between the serum ferritin level as a measure of iron store and the maximum increase in serum iron during a 3‐h test (Cmax), in the elderly as well as in the healthy adult group. A decision limit of 80 μg/dL forCmaxis a good discriminant between absent (serum ferritin <20 μg/L) and adequate body iron stores. Sixty‐eight per cent of the patients with a serum ferritin level <20 μg/L but virtually none of the ACD patients, non‐anaemic elderly inpatients with normal serum ferritin levels and healthy adults had aCmaxlevel >80 μg/L. Although further investigation is needed before the OIAT can be recommended as a valuable test for evaluating iron absorption, predicting mild iron deficiency and differentiating between different categories of anaemia, it seems worthwile that more effort should be done to validate this simple and safe test.
Abnormal bone turnover in monoclonal gammopathy of undetermined significance: analyses of type I collagen telopeptide, osteocalcin, bone‐specific alkaline phosphatase and propeptides of type I and type III procollagens
Authors: Vejlgaard T. , Abildgaard N. , Jans H. , Nielsen J. Lanng , Heickendorff L.
Published: July 2010

Abstract: The main difference between monoclonal gammopathy of undetermined significance (MGUS) and multiple myeloma (MM) is the presence of lytic bone destructions in the latter. About 20% of MGUS patients develop MM, and histomorphometric studies have shown disturbed bone turnover rates in some of these patients. This study was performed in order to evaluate whether serum analyses of the C‐terminal telopeptide of type I collagen (ICTP), as a reflector of bone degradation, and of osteocalcin, bone‐specific alkaline phosphatase (bAP) and the C‐terminal propeptide of type I procollagen (PICP), as markers of bone formation, might give information on disturbancies of bone metabolism in MGUS. Furthermore, serum N‐terminal propeptide of procollagen III (PIIINP) might give information on disturbances in collagen III metabolism in the bone marrow. In the 35 patients examined, serum ICTP was elevated in 12 patients (34%), serum PIIINP elevated in 6 patients (17%), serum osteocalcin elevated in 11 patients (31%), serum bAP elevated in 6 patients (17%), and serum PICP elevated in 4 patients (11%). Serum ICTP correlated significantly with PIIINP (r=0.72, p<0.001), and with serum osteocalcin (r=0.57, p<0.001) and serum bAP (r=0.51, p=0.002). These findings indicate disturbancies of bone turnover and affected collagen metabolism in some MGUS patients. Follow‐up observation may reveal any prognostic value of these findings.
Autoimmune haemolytic anaemia in patients with chronic lymphocytic eukaemia treated with 2‐chlorodeoxyadenosine (cladribine)
Authors: Robak T. , Błasińska‐Morawiec M. , Krykowski E. , Hellmann A. , Konopka L.
Published: July 2010

Abstract: Autoimmune haemolytic anaemia (AIHA) is one of the major complications in chronic lymphocytic leukaemia (CLL). Treatment with alkylating agents and the adenosine analogue, fludarabine, might trigger the development of AIHA in CLL patients despite the reduction of leukaemic clone. The influence of 2‐chlorodeoxyadenosine (2‐CdA) on AIHA in patients with CLL is undefined so far. In a group of 114 patients treated at our clinics with this agent, AIHA with direct antiglobulin test (DAT) positively was observed in 25 (21.9%) patients. In 23 patients AIHA was noticed before the starting of 2‐CdA and in 2 patients DAT became positive after 2‐CdA treatment. In 6 patients the drug caused complete resolution of haemolysis and DAT became negative. Eight patients exhibited partial resolution of haemolysis with significant improvement in haemoglobin level but DAT test remained positive. In 11 patients there was no response to 2‐CdA in relation to autoimmune haemolysis. Two patients with no previous history of haemolysis developed AIHA after 5 and 6 courses of 2‐CdA therapy and their DAT became positive 1–2 months after the last course of the drug. One of them entered severe haemolytic crisis and severe thrombocytopenia and died because of haemorrhagy to the central nervous system. In the other patient AIHA was controlled by steroids and chlorambucil treatment. Our study indicates that 2‐CdA may suppress autoimmunohaemolytic process in some patients with CLL and trigger the development of AIHA in others.
High levels of the DNA‐binding activity of E2F in adult T‐cell leukemia and human T‐cell leukemia virus type I‐infected cells: possible enhancement of DNA‐binding of E2F by the human T‐cell leukemia virus I transactivating protein, Tax
Authors: Mori Naoki
Published: July 2010

Abstract: Transcription factor E2F binds to cellular promoters of certain growth‐ and cell cycle‐controlling genes and forms distinct heteromeric complexes with other nuclear proteins. It has therefore been proposed that E2F is involved in cellular proliferation control. Human T‐cell leukemia virus type I (HTLV‐I) is an etiological agent of adult T‐cell leukemia (ATL). We show here by mobility‐shift assay that E2F‐containing DNA‐binding complexes were detected in HTLV‐I‐infected T‐cell lines and leukemic cells obtained from ATL patients but not in an uninfected T‐cell line, Jurkat, and normal peripheral blood mononuclear cells. The Tax protein, encoded by HTLV‐I, is a potent transcription activator of viral and several cellular genes. We demonstrate that expression of Tax can induce the E2F‐containing DNA‐binding complexes in Jurkat T cells. Thus, Tax, through enhancement of the DNA‐binding activity of E2F, may be capable of regulating cellular gene expression implicated in the proliferation and transformation of T cells in ATL. This activity may be relevant to the mechanisms whereby HTLV‐I which does not contain oncogenes induces neoplasia.
Biosynthesis, processing and sorting of neutrophil proteins: insight into neutrophil granule development
Authors: Gullberg Urban , Andersson Elinor , Garwicz Daniel , Lindmark Anders , Olsson Inge
Published: July 2010

Abstract: Neutrophil granulocytes are specialized phagocytic cells that carry a collection of granules for regulated secretion, each with distinct constituents. The granules can be classified as azurophil (primary), developed first, followed in time by specific (secondary) granules gelatinase granules, and secretory vesicles. Stage‐ and tissue‐specific transcription factors govern the successive expression of genes for granule proteins to allow storage of the gene products in these organelle categories whose packaging is separated in time. Many of the granule proteins, in particular those of the heterogeneous lysosome‐like azurophil granules, are subject to extensive post‐translational proteolytic processing into mature proteins, most commonly as a post‐sorting event. A selective aggregation of proteins destined for storage in granules, as discussed in this review, would facilitate their retention and eliminate a need for distinct sorting motifs on each granule protein. Aggregation of granule proteins, that are often cationic, would be assisted by the anionic serglycin proteoglycans present in neutrophils. The antibacterial granule proteins can serve as models for antibiotics and some of them possess a potentially useful therapeutic ability to bind and neutralize endotoxin. Because aberrant expression of transcription factors regulating the synthesis of granule proteins is often found in leukemia, the clarification of mechanisms regulating the timed expression of granule proteins will shed light on the maturation block in myeloid leukemias.