bluuebunny/crossref_metadata_2025 huggingface dataset:
Exocytosis of zymosan particles by human phagocytes
Authors: Bjerknes Robert
Published: August 2010

The kinetics of human leucocyte phagocytosis and exocytosis of fluorescein‐isothiocyanate (FITC)‐labelled zymosan particles were studied by flow cytometry (FCM). The leucocytes rapidly associated with zymosan particles, and internalization was confirmed by a fluorescence quenching technique. For incubation periods longer than about 60 min, exocytosis of ingested particles was observed. All human phagocytes ejected zymosan particles. The rate of exocytosis was about 1 particle per phagocyte per h, and was independent of the number of internalized particles. Exocytosis was dependent on temperature and glucose, but did not require Ca++ and Mg++ ions. Phagocytosis and exocytosis occurred concurrently, and phagocytosis accelerated ejection of previously internalized particles. The exocytosed particles were partially degraded by the phagocytes, and rephagocytosis of ejected zymosan particles was slower than the uptake of control particles. Phagocytes undergoing exocytosis remained intact during the 210 min examined. The findings indicate a heretofore neglected vector in the interaction of prey and phagocytosing cells.
Introduction to the Sexual Consequences of Hemophilia
Authors: Levine S B
Published: August 2010

The sexual consequences of chronic pediatric disease are only now beginning to be studied. Most clinicians do not yet have the conceptual tools to thoroughly assess the sexual life of young adult hemophiliacs. This paper presents five major landmarks of sexual development: gender identity, orientation, intention, function (desire, arousal, orgasm, emotional satisfaction) and the enabling relationship. Several hypotheses about the impact of hemophilia are described. First, the illness identity of the person with chronic disease threatens to supercede the masculine gender identity. Second, an unappreciated high prevalence of orientation and intention problems lurk behind the high frequency of heterosexual “shyness”. Suggestions to maximize the sexual developmental potential of young hemophiliacs are provided based on our current concepts of child development and sexual development in particular.
The Management of Haemophiliacs who have Antibodies to Factor VIII
Authors: Rizza C R
Published: August 2010

The management of haemophiliacs with factor VIII antibodies is still one of the major problems in the treatment of haemophilia in spite of our greater understanding of the nature and mode of action of those antibodies.

The approach to treatment varies from centre to centre and may consist of factor replacement with human or porcine factor VIII, administration of prothrombin complex concentrates, plasmapheresis, immunosuppressive drugs or different combinations of the above.

At Oxford we have relied entirely on human factor VIII replacement for the management of acute bleeding. In twenty‐four patients receiving on demand therapy for 2–10 years we have details of amounts of factor VIII used, frequency of dosage and levels of factor VIII antibody. Eight patients with low levels of antibody have lost their antibody. In 7 patients with higher levels there has been a reduction in anamestic response and level of antibody in the blood. In the remaining patients some of whom are infrequently treated there has been little change in the antibody response.

The above findings are described in the light of other regimens of therapy employing high or low dosage factor VIII replacement.
Overcoming Factor VIII Inhibitors: A Possible New Approach
Authors: Barrowcliffe T W , Kemball‐Cook G , Gray E
Published: August 2010

Treatment of haemophiliacs with antibodies to Factor viii is still a major clinical problem, neither high‐dose Factor viii nor Factor ix concentrates being completely effective. An alternative approach was suggested by the finding that Factor viii, when complexed with phospholipid and Factor IXa, was protected from antibody attack, and that the major protective effect was provided by the phospholipid.

The protective effects of various phospholipids were assessed by addition to Factor viii concentrates, incubation with human antibodies to VIII:C and measurement of residual clotting activity in three different assay systems. The most active material was a home‐made extract of bovine brain, prepared by a modified Folch method. Of three commercial lipid preparations manufactured for intravenous injection, one was inactive and the other two were only active at high concentration.

Studies with purified phospholipids showed that phosphatidyl serine (PS) was the most active fraction. Phospholipid analyses showed that the three intravenous lipid preparations all contained less than 10% PS, whereas the bovine brain extract contained 20–25%.

These studies show that Factor viii can be protected from antibody attack by addition of phospholipid extracts which are rich in PS, and suggest that a Factor viii product containing a suitable phospholipid could be of benefit to haemophiliacs with inhibitors.
Use of HBV Vaccine in Hemophiliacs
Authors: Gazengel C. , Courouce A.M. , Torchet M.F. , Kremp O. , Brangier J. , Brechot C. , Degos F.
Published: August 2010

Patients with coagulation defects received three monthly injections of vaccine against Hepatitis B surface antigen (HEVAC B from Institut Pasteur Production, France). We now report preliminary results obtained in 26 hemophiliacs and in 7 patients with other coagulation defects. Very few side effects were observed. Immune response (anti HBs antibody 5–50 mIU/ml) was obtained in 32 patients (97 percent). The geometric mean of anti HBs titer one month after the 3rd injection of vaccine was 2,014 mIU/ml (95 percent confidence interval 1,195 to 3,331 mIU/ml) in 28 patients. Transaminase activity (ALT) was assayed in 31 patients: in 16 it was within normal limits (> 43 IU/L), a subsequent ALT increase was observed in 4. In the 15 other patients ALT was high at the beginning and did not change during the follow‐up. Neither HBsAg nor anti HBc (except for one patient with a low anti HBc level after F IX infusion) was detected. HBV‐DNA assay in the serum performed in patients with abnormal ALT activity, was negative.
Liver Disease and Pathology in Hemophilia
Authors: Colombo M , Rumi M G
Published: August 2010

Hemophilic patients given replacement therapy with clotting factors are frequently exposed to hepatitis viruses or viral products. For the large percentage of these patients with serological evidence for past or current hepatitis B infection, the prevalence of clinically recognizable hepatitis is surprisingly low. Several prospective studies of asymptomatic patients with hemophilia have revealed that many of them have prolonged, mild‐to‐moderate elevations of serum transaminase. Liver biopsies taken from such patients provided unequivocal evidence for the existence of chronic hepatitis. Current studies of hemophiliacs with chronic hepatitis indicate that HBsAg‐positive patients may develop severe forms of liver disease, while markers of intrahepatic delta agent replication have been found only in patients with chronic active hepatitis or cirrhosis. On the contrary nonA nonB chronic hepatitis appears to be milder than expected from studies of patients undergoing open‐heart surgery. The giving of factor concentrates during the first years of life seems to be important for development of liver disease. However, the nature of the infection (type B and delta) rather than the cumulative life‐time exposure to concentrates seems to be the important element for determining the severity of the hepatic lesions.
Alteration of T‐Lymphocyte Subpopulations in Children, Youth and Adults with Hemophilia A
Authors: Wolff L J , Borzy M , Kamoun M , Lovrien E W
Published: August 2010

Thymic processed lymphocytes subpopulations were assayed in 32 healthy patients with hemophilia A and no inhibitor. Commercial concentrates were infused at various frequencies. Monoclonal antibodies to OKT3, OKT4 and OKT8 were measured and compared to age controls. A progressive decrease in percent OKT4 cells, a progressive increase in percent OKT8 cells and gradual decline in the T4/T8 ratio were noted with increase in the patients' age. A correlation between the amount of concentrate received and alteration in T cell subsets was observed in 4 children only. These early and progressive alterations in the T‐lymphocyte subsets suggest early immunocompromise and warrant close patient surveillance and immunologic scrutiny.
Immunological Dysfunction and Persistent Lymphadenopathy in Patients with Classic Hemophilia
Authors: Tsoukas C , Gervais F , Fuks A , Guttmann R D , Strawczynski H , Shuster J , Gold P
Published: August 2010

Thirty‐four asymptomatic heterosexual adults with severe classic hemophilia without inhibitors receiving more than 40,000 units of factor VIII concentrate per year were studied and compared to 10 heterosexuals receiving cryoprecipitate and to 22 age matched controls. Anergy was noted in 68 percent of the patients treated with factor VIII concentrates, 57 percent of those treated with cryoprecipitate and 5 percent of the controls. The lyophilized factor VIII treated group had a significant elevation of serum IgG, decreased mitogen responses, high nonstimulated background activity, and altered T cell subsets with an increase in total T lymphocytes, T suppressor cells and decreased Th/Ts ratios.

Four of the patients with cellular immune dysfunction developed fatigue, night seats, dyspnea, weight loss, neuropathic pain and rashes. They had diffuse, peripheral, non‐tender lymphadenopathy. Two had splenomegaly. All four were anergic, had inverted Th/Ts ratios, with an increase in suppressor cells. Three had lymph node biopsies which demonstrated reactive follicular hyperplasia. All had elevated serial titers to EBV. The systemic symptoms resolved as these titers returned towards normal. The lymphadenopathy is persistent although all four patients have gained weight and feel well. The persistent lymphadenopathy syndrome in hemophilia is a distinct entity which occurs in a subset of hemophiliacs with severe cellular immune dysfunction.
Alternatives to Financing
Authors: Aledort L M
Published: August 2010

Comprehensive care for hemophilia patients has clearly demonstrated that it is cost effective and offers the patient a better, more active life. However, the cost of underwriting the core personnel has not been adequately addressed or resolved.

Alternatives are presented which include a) federal or state support, b) third party reimbursement mechanisms, c) liaisons with blood programs and, d) cost overrides on product.
Innovative Alternatives to Human Factor VIII
Authors: Tuddenham E.G.D.
Published: August 2010

Despite massive expenditure of effort and money in fractionation of human blood plasma, it is clear that the end‐product factor VIII is deficient in quantity and its quality seriously compromised by infectious agents both known and unknown. These problems may not be soluble by ever greater blood collection drives and ever more stringent process controls. The average fractionation puts only 20 per cent of donated factor into the bottle of concentrate. Are radical new methods of purification needed?

Meanwhile, the vast majority of our potential resources of factor VIII and IX, free of all known human pathogens are thrown away. In the United Kingdom last year, over 600,000 million units of sheep, pig and cow factor VIII were washed down the abbatoir gutters or consumed as black pudding. Already, some haemophiliacs with inhibitors are successfully treated with highly purified porcine factor VIII. Could new purification methods and chemical modifications make this natural abundance of animal factor VIII available to treat all haemophiliacs?

Genetically engineered factor VIII made by cultured cells, pure and in unlimited quantities is a theoretical possibility which stands as a ‐ perhaps the ‐ supreme challenge to biotechnology today. Undaunted, several groups have taken up this quest. Huge obstacles bar the way but like Everest it attracts our highest endeavours because it is there.

Finally, what thoughts should we entertain of gene therapy? “Ah, could we not seize this sorry scheme of things entire and mould it closer to the heart's desire?” There have been false dawns but recent progress on gene insertion and control raise more than a glimmer of fresh hope.
The National and International Layman's Role in the Patient's Treatment
Authors: Voulis C
Published: August 2010

Laymen play a severe role in the hemophiliacs treatment. Laymen f.i., can help hemophiliacs in their home treatment, can be members of a voluntary personnel at the Society's Office or at the Hemophilia Centre, can be Blood Donors, can prepare educational/professional programs for the hemophiliacs etc. For a better use of the Laymen it is proposed: Programs of public information about the fundamental causes of hemophilia, Advisory Committees with a great Laymen's participation, training of Laymen (especially for the developing countries). Finally we must note that no one program or Laymen's plan could be successful unless it gains the acceptance of the hemophiliacs themselves.
METHODOLOGY OF THE TWO‐STAGE ASSAY OF FACTOR VIII (VIII:C)
Authors: Barrowcliffe Trevor W.
Published: August 2010

ABSTRACT:  The principle of the two‐stage assay, the preparation of reagents and the performance of the assay are described in detail. Modifications introduced by various authors are discussed, including a simplified version designed for automation, and a chromogenic assay. Finally, examples are given of the methodology used in our laboratory.
THE CONTROL AND STANDARDIZATION OF FACTOR VIII
Authors: Aronson David L. , Thomas Duncan P.
Published: August 2010

ABSTRACT:  Factor VIII concentrates have been used increasingly over the past decade for the treatment of patients with classical haemophilia. To ensure broad comparability between the potency estimates of therapeutic concentrates prepared by different manufacturers, a Factor VIII standard is required.

The principles of biological standardization are discussed, particularly as they apply to the standardization of Factor VIII, both in plasma and concentrates.

The role of the WHO International Standards is considered, as well as the importance of secondary standards. The control of Factor VIII concentrates by the regulatory authorities in the U.S.A. and the U.K. is also discussed. It is concluded that optimal control of Factor VIII concentrates requires a continuing interchange between producers, regulators and users.
Characterization of transglutaminases in normal and malignant human leukocytes
Authors: Berntorp Erik , Seiving Birgitta , Stenberg Pal
Published: August 2010

Using the fluorescent activity staining procedure, transglutaminases in human monocytes, granulocytes and lymphocytes have been characterized with respect to agarose gel electrophoretic mobility and thrombin dependence. A thrombin dependent transglutaminase was found in concanavalin A stimulated peripheral monocytes. The electrophoretic mobility of this zymogen and of platelet and plasma factor XIII was similar. With stimulated peripheral lymphocytes a similar pattern was observed. However, the potential transglutaminase activity in the lymphocyte factor XIII was lower than that in monocytes. Similar results were obtained when lymphocytes from chronic myeloid and chronic lymphocytic leukaemia patients were examined. Quantitatively, the transglutaminase activity in the leukaemic cells was estimated to be lower than the activity in normal cells. With respect to agarose gel electrophoretic mobility, a similar transglutaminase was found in concanavalin A stimulated human peripheral granulocytes. Although electrophoretically clearly separated from tissues transglutaminase, the granulocyte enzyme did not seem to require thrombin for activation.
Immune phenotype heterogeneity in AML
Authors: Öst Å. , Christensson B. , Andreasen R. , Hast R. , Lagerlöf B. , Reizenstein P. , Trowbridge I. S. , Biberfeld P.
Published: August 2010

Blood cells from 46 patients with acute myeloid leukaemia were studied for expression of various surface markers, defined by a panel of 12 monoclonal antibodies and the expression of Fcγ receptors. Corresponding studies were done on normal bone marrow cells. Antibodies which bound to leukaemic cells in high frequencies were those which most frequently also bound to normal bone marrow cells. Immunophenotypic analysis revealed a marked antigenic heterogeneity in AML, also evident within single FAB subclasses. However, leukaemic cells of FAB subclass M1 significantly more often expressed HLA class I antigen than those of FAB subclass M5a, whereas Fcγ receptors which were expressed only on a few cells in M5a, were increasingly frequent on leukaemic cells of M1‐M2, M4, and M5b leukaemias.

The frequency of cells reacting with the monoclonal antibody T50/12,11,2 was related to the complete remission rate of the patients. Patients with high frequencies of cells reacting with this antibody had a better complete remission rate than patients with fewer cells binding to this antibody.

The immunophenotypic heterogeneity an AML may reflect a great biological variability of this disease. This variability may be of importance for the classification and treatment of AML.
A method for quantitation of H+‐ion activity relating to packed erythrocytes
Authors: Torp. H. E.
Published: August 2010

To try to contribute new information on the problem concerning red cell destruction in the spleen, a new method which makes it possible indirectly to quantitate H+‐ion activity relating to packed erythrocytes, has been devised.

The method is based on the chemical reaction:

5 I– + I O–3 + 6 H+ ⇄ 3 I2 + 3 H2O

When blood is added to a solution of iodate and iodine in excess, the amount of iodine formed depends on the H+‐ion quantity offered by the blood. The erythrocytes primarily act as H+ donors, secondarily as iodine acceptors. Acetone added primarily liberates iodine from the erythrocytes and subsequently acts as a solvent. The amount of dissolved iodine was determined by photometry. The method makes it possible to quantitate the H+‐ion activity relating to packed erythrocytes in microvolumes of blood.
Quantitation of H+‐ions relating to packed erythrocytes in blood from the spleen and the inferior vena cava in rats
Authors: Torp. H.E.
Published: August 2010

Red cell destruction in the spleen may to some extent depend on a possibly increased H+‐ion activity relating to packed red blood cells in the spleen. A number of blood samples from the spleen and inferior vena cava in rats were examined with a method for determination of H+‐ion activity relating to packed erythrocytes devised by the author. The H+‐ion activity was invariably greater in the blood samples from the spleen. Blood samples from the spleen were obtained with a technique avoiding profuse bleeding. Haemolysis in the spleen on the basis of increased H+‐ion cone, are discussed.
A methodological evaluation of 14 controlled clinical trials in myelomatosis
Authors: Birgens Henrik S. , Hansen Ole Paaske , Clausen Nielsaage Tøffner
Published: August 2010

A methodological evaluation of the 14 randomized clinical controlled trials so far published on myelomatosis was performed. A quite large heterogeneity was observed among the trials, especially in their presentation of patients included and in the criteria applied for therapeutic response. Additionally, none of the studies accounted for the randomization procedure — and particularly whether the randomizations were performed consecutively. 19 comparisons of the effectiveness of various drugs or drug combinations were performed and in 12 of these an insignificant result was obtained. 2 of these with more than 25% probability have overlooked a beneficial effect of one of the regimens of more than 50% and 6 trials of more than 25%. It is stressed that these situations will arise when sample sizes that are too small are applied in clinical trials. It is concluded that internationally accepted criteria for diagnosis and therapy response are needed. Additionally, a more thorough statistical planning, prior to initiation of a trial, and a more extensive analysis of the results would be helpful.
Cerebrospinal fluid ferritin in patients with leukaemia and malignant lymphoma
Authors: Milman Nils , Vig Louise , Pedersen Nils Strandberg , Olsen Tom Skyhøj
Published: August 2010

To evaluate whether cerebrospinal fluid (CSF) ferritin could be of diagnostic value in haematological malignancies with central nervous system (CNS) involvement, the ferritin concentration was measured in 21 patients with acute leukaemia and lymphoma. Of the 17 patients without CNS involvement, 16 had CSF ferritin values in the normal range (2–7 μg/l); 1 patient had an elevated value, probably due to blood contamination in connection with a very high serum ferritin level. 4 patients had tumour invasion of the CNS indicated by the presence of blastic cells in the CSF; CSF ferritin levels in these patients were likewise in the normal range. There was no difference between CSF ferritin values in patients with and without CNS involvement. With the present assay, measurement of CSF ferritin appears to be irrelevant in the evaluation of CNS invasion in haematological malignancies.
The dynamic morphology of the transferrin‐transferrin receptor system in human leukaemia/lymphoma cell lines and its relation to iron metabolism and cell proliferation
Authors: Forsbeck Klas , Nilsson Kenneth
Published: August 2010

Some functional and morphological aspects of the transferrin‐transferrin‐receptor (Tf‐TfR) system were investigated in 12 human haematopoietic tumour cell lines. The iron uptake ability, studied by 59Fe labelling, varied considerably between individual tumour cell lines. The studies of the morphology of the Tf‐TfR system by fluorescence based methods using labelling by FITC‐Tf, and by indirect immunofluorescence (OKT9 anti‐TfR monoclonal antibody) demonstrated that the iron uptake was the result of receptor‐mediated endocytosis (RME) of the Tf‐TfR complex. The RME was better developed in the haemoglobin synthesizing K‐562 cells than in the monocytic U‐937 cells, suggesting that the intracellular iron processing capacity differs from cell type to cell type.

The efficiency of iron‐uptake, transport and storage was related to cell growth. Inhibition of growth by increasing cell densities or by drug treatments (phorbol ester TPA and difluoromethylornithine DFMO) was thus accompanied by a decrease in surface Tf binding, in cellular Tf handling capacity and in iron accumulation. TPA induced a redistribution of the TfR‐pool to the intracellular space as demonstrated by morphology.
Glutathione peroxidase activity and reduced glutathione content in erythrocytes of patients with chronic renal failure.
Authors: Seth R. K. , Saini A. S. , Aggarwal S. K.
Published: August 2010

Erythrocytes from 18 patients with chronic renal failure (CRF) and 10 healthy subjects were examined with respect to glutathione peroxidase (GSH‐Px) activity and reduced glutathione (GSH) contents. The activity of GSH‐Px and GSH content were found to be lower in RBC from CRF patients as compared with normal RBC. These reduced levels of GSH and GSH‐Px in the red cells of uraemic patients may predispose the cells to oxidative damage.
T‐cell abnormalities in patients with idiopathic thrombocytopenic purpura: The presence of OKT4+8+ cells
Authors: Mizutani Hajime , Katagiri Shuichi , Uejima Kazuo , Ohnishi Mitsunobu , Tamaki Toshiharu , Kanayama Yoshio , Tsubakio Tadahiro , Kurata Yoshiyuki , Yonezawa Takeshi , Tarui Seiichiro
Published: August 2010

40 patients with idiopathic thrombocytopenic purpura (ITP) were studied for T‐cell abnormalities using a panel of monoclonal antibodies (Mc Abs). These patients showed a tendency to have a decreased OKT4+: OKT8+ ratio compared with normal subjects and a significant increase was observed in OKT10‐positive cells in ITP patients. Further analyses were made utilizing McAbs of different subclasses in combination (BMA040 mouse IgG1 McAb, helper/inducer T and BMA081 mouse IgG2 McAb, suppressor/cytotoxic T). An increased proportion of cells reactive with both BMA040 and BMA081 McAbs (double‐labelled cells) was demonstrated in patients with ITP compared with the normal controls. Furthermore, there was an inverse correlation between the proportion of “double‐labelled cells” and the platelet counts. These data suggest that abnormalities of the T‐cell subsets in the peripheral blood may play an important role in the pathogenesis of ITP.
PF 4 versus βTG as evidence for platelet activation in myeloproliferative disorders
Authors: Luzzatto G. , Fabris F. , Mazzucato M. , Girolami A.
Published: August 2010

19 patients with MPD have been studied. As described in normals, an age‐related increase in β thromboglobulin (βTG) release is observed. Such release, however, is greater in patients with myeloproliferative disorders (MPD). MPD seem therefore to cause platelet activation, allowing an earlier and more evident manifestation of physiologic ageing phenomena. PF 4 levels are near zero both in controls and patients, regardless of platelet number. This suggests that increased levels of PF 4 represent only a laboratory artifact, caused by platelet activation in vitro. Mean ability in producing thromboxane B2 (TxB2) is increased, but is perfectly normal in patients with normal platelet count and decreased in 3 thrombocythaemic patients, who seem to present an increased thrombotic risk. TxB2 is reduced almost to zero by the administration of aspirine plus dipyridamole; contrarily, all other parameters were unaffected, either by such drugs or by AD 6, a new coumarin derivative with antiplatelet properties.
Speed of relapse in children with acute lymphoblastic leukaemia
Authors: Bareford D. , Bailey C. C. , Robinson E. A. E.
Published: August 2010

16 children with acute lymphoblastic leukaemia (ALL) who relapsed on maintenance treatment between September 1978 and December 1981 were studied. Their previous bone marrow aspirates were reviewed to determine the first evidence for marrow relapse and the subsequent rate of evolution of the disease. 60% of children had bone marrow evidence of unsuspected relapse at 8 wk, and 40% had evidence at 12 wk before clinical or peripheral blood relapse occurred. Early detection of relapse will prevent continued use of ineffective maintenance chemotherapy and may also reduce morbidity during subsequent induction therapy. Regular 8‐weekly marrow aspirates are therefore recommended during first remission of ALL in children with a histocompatible sibling who would be eligible for bone marrow transplantation in second remission.
Cutaneous pre‐B leukaemia cells with a T‐lymphocyte antigen (T4)
Authors: Nakamura Hirotoshi , Koike Takao , Itoh Kuniaki , Maruyama Naoki , Tomioka Hisao , Yoshida Sho
Published: August 2010

A patient with acute lymphocytic leukaemia (ALL) is described in whom the cells originated in the pre‐B cell lineage but showed an aberrant expression of T4 antigen. A B‐cell origin of the leukaemic cells was indicated by reactivity for several monoclonal antibodies specific for the B‐cell lineage and by cytoplasmic staining with FITC‐labelled anti human μ chains.
Aseptic bone/bone marrow necrosis in leukaemia
Authors: Vesterby Annie , Jensen O. Myhre
Published: August 2010

Aseptic (avascular) bone/bone marrow necrosis (ABN) was found in 21 leukaemic patients during a period of 14 yr, with a frequency of 6.5% in autopsied leukaemic patients, and with the highest frequency in ALL. 8 patients (38%) complained of bone pain. X‐ray examination was carried out in 7 of these with a positive result in 3 (43%). Only 1 patient had elevated S‐Alkaline phosphatase. Leucocytosis was found in 12 patients. Only 1 patient (ALL) received steroid in a rather high dose during 6 weeks. At autopsy ABN was found localized to the femur in all patients and 2 patients also had ABN in other bones. Post‐mortem X‐ray examination demonstrated changes in 8 of 15 cases (53%), with osteolysis in 6 and sclerosis in 2. 19 patients had had recent ABN with some fibroblast proliferation. In 4 of these, appositional bone formation had started. 2 patients had sclerosis as the only change.

The pathogenesis of ABN is not known; an important factor, however, may be ischaemia due to vascular obstruction.
Circulating immune complexes in myelofibrosis
Authors: Hasselbalch Hans , Nielsen Henrik , Berild Dag , Kappelgaard Ellen
Published: August 2010

17 patients with idiopathic myelofibrosis were studied for the occurrence of circulating immune complexes (IC), using a polyethylene glycol complement consumption and a polyclonal rheumatoid factor inhibition assay. In 13 patients complement C3d was determined by rocket immunoelectrophoresis. Circulating IC were detected in 6 patients and were primarily found in patients with short duration of disease from time of diagnosis. The median duration of the disease in IC‐positive patients was 4 months, compared to 12 months in the IC‐negative group (P<0.05). 9 of the 13 patients investigated had increased levels of plasma C3d. However, there was no correlation to the occurrence of IC. It is concluded that circulating IC may take part in an immune‐mediated bone marrow damage. This may involve deposition of IC in the bone marrow with secondary inflammation responsible for the development of bone marrow fibrosis.
Treatment of therapy‐resistant Sézary Syndrome with Cyclosporin‐A: Suppression of pruritus, leukaemic T cell activation markers and tumour mass
Authors: Tötterman T. H. , Scheynius Annika , Killander A. , Danersund A. , Alm G. V.
Published: August 2010

A patient with Sézary's syndrome resistant to conventional therapy was successfully treated with Cyclosporin‐A (CyA) for 14 months. Pre‐treatment in vitro studies showed that the leukaemic T cells were sensitive to therapeutic concentrations of CyA. The patient's pruritus disappeared promptly after 2 d of treatment. The positive effect of CyA on the pruritus was related to the dose and plasma concentrations of the drug. Analysis of leukaemic cell surface markers using monoclonal antibodies showed that episodes of pruritus were correlated with the appearance of cells expressing the T cell “activation” markers interferon‐ gamma (IFN‐λ) and HLA‐DR. This indicated that CyA may control pruritus by suppressing the production/release of lymphokines. Immunohistochemical staining of repeated skin biopsies demonstrated a reduction of infiltrating T cells. Clinically, this was correlated with an improved skin status and a partial regress of palpable lymph node size. Apart from an initial reversible drop in the circulating helper/suppressor T cell ratio, the number of circulating Sézary cells was unaltered during CyA treatment. After 5 months, higher doses of CyA were needed to control symptoms, and this was correlated with partial drug resistance also in vitro.
Density of granulomonocytic colony‐forming cells (GM‐CFC's) in myelofibrosis
Authors: Charbord P. , Neel H.
Published: August 2010

The circulating GM‐CFC density‐distribution profile was studied in 19 cases of myelofibrosis with myeloid metaplasia. The distribution profile for the population studied appeared to be similar to that of normal bone marrow GM‐CFC's but shifted toward lower densities in comparison with normal circulating GM‐CFC's. For individual patients, it appeared that the greater the circulating blood GM‐CFC's concentration, the lower the mean density. It appears likely that the premature release into blood of the GM‐CFC's in myelofibrosis is a property of the neoplastic haematopoietic stem cell clone at the core of the disease.
Characterisation of a new cell line (CJ18) from a patient with ‘hairy’ cell leukaemia
Authors: Skinnider Leo F. , Ha Tarn , Bergen Susan , Wang H. C.
Published: August 2010

A cell line (CJ18) has been established from the peripheral blood of a patient with hairy cell leukaemia (HCL) in a leukaemic phase. They grow slowly in large clumps with a doubling time of 3–4 d. 8% show positivity for surface immunoglobulin G and a small percentage (5%) are positive for intracytoplasmic immunoglobulin. They are B1, Ia1 positive and CALL, TdT and OKM1 negative. Although they are Epstein Barr Virus Nuclear Antigen (EBNA) positive they have several features not found in other EBNA positive B lymphoblastoid cell lines which suggest they may be derived from the patient's leukaemic hairy cells. They are strongly positive for tartrate resistant acid phosphatase, esterase positive, contain numerous lysosomes and are able to phagocytose sheep red blood cells after exposure to tetradecanoyl‐12, 13‐phorbol acetate (TPA). Following exposure to retinoic acid and TPA they adhere to plastic with numerous slender processes, a feature seen in HCL cells.
High expression of lymphocyte surface immunoglobulin in chronic lymphocytic leukaemia may be a bad prognostic sign
Authors: Berntorp Erik , Zettervall Olle
Published: August 2010

Lymphocytes from 27 patients with B‐type chronic lymphocytic leukaemia (CLL) were tested with immunofluorescence for expression of surface membrane immunoglobulin (SmIg) and the B cell antigen 7420. When age at time of testing, sex, clinical stage according to Binet, disease progression according to Levin and percentage of SmIg and 7420 antigen‐expressing cells were used as explanatory variables for survival in a linear logistic regression model, only the SmIg variable was significant (p < 0.025). A high proportion of SmIg bearing cells (> 61%) was associated with short survival, large tumour mass and aggressive disease. The tentative conclusion is drawn that the differentiation stage of the tumour cells in CLL determines the clinical course of the disease and is inversely correlated to the SmIg expression.
Effect of D‐penicillamine on human granulopoiesis in vitro
Authors: Aymard J. P. , Herbeuval M. , Netter P. , Legras B. , Streiff F. , Herbeuval R.
Published: August 2010

The effect of D‐penicillamine on human bone marrow granulocyte‐monocyte precursor cells (CFU‐C) is studied in vitro. Bone marrow samples were obtained from 47 donors (40 patients and 7 healthy volunteers) and cultured in semisolid agar in the presence of various concentrations of the drug. We found an inhibitory, possibly dose‐dependent, effect of D‐penicillamine on colony formation. The pathophysiological mechanism and the clinical relevance of this effect remain poorly understood. These findings, however, emphasize the need for careful monitoring of the granulocyte counts during D‐penicillamine therapy.
Coproporphyrinogen oxidase activity and porphyrin concentrations in peripheral red blood cells in hereditary sideroblastic anaemia
Authors: Pasanen A.V.O. , EklöUf M. , Tenhunen R.
Published: August 2010

The activity of coproporphyrinogen oxidase and the concentrations of coproporphyrin and protoporphyrin (measured by HPLC) in peripheral red blood cells were established in 2 families with different types of hereditary sideroblastic anaemia. 2 males and 4 females were members of a family with an X‐chromosome‐linked and pyridoxine‐responsive HSA, and 3 females were members of another family where the mode of inheritance is not clear and where pyridoxine did not produce a haemotological response. Coproporphyrinogen oxidase activity was normal in 8 of 9 patients and slightly decreased only in 1 patient. All patients had normal red cell coproporphyrin concentrations, but red cell protoporphyrin concentration was decreased in 4 patients.

These findings indicate that in vivo haem synthesis was not impaired at the step of coproporphyrinogen oxidase, hence enzymatic defects in earlier steps of haem synthesis are more evident. Earlier suggestions of impaired haem synthesis at this level, based on observed increased concentrations of coproporphyrin in peripheral red blood cells might be explained by the use of unspecific methods.
Increased leucocyte alkaline phosphatase and transcobalamin III in chronic myeloid leukaemia associated with lithium therapy
Authors: Moreb Jan , Hershko Chaim
Published: August 2010

A 38‐year‐old woman developed chronic myeloid leukæmia after 2 years of lithium carbonate therapy. A peculiar feature of her leukaemia, as well as of the 5 patients previously reported in whom CML has developed in the course of lithium therapy, was the unusually high degree of granulocyte maturation manifested in normal leucocyte alkaline phosphatase (LAP) score and, in 1 case, selective increase of transcobalamin III. Although a cause and effect relation between lithium therapy and CML has not yet been established, in view of the stimulatory effect of lithium on granulocyte proliferation, such treatment should be avoided in patients with established myeloproliferative disorders, or in patients at high risk of developing leukaemia.
Evaluation of neutrophil alkaline phosphatase (NAP) activity: Untreated myeloid leukaemia, lymphoid leukaemia and normal humans
Authors: Bendix‐Hansen Knud , Helleberg‐Rasmussen Inge
Published: August 2010

Neutrophil alkaline phosphatase (NAP) activity was estimated in 50 healthy humans and 89 patients with leukaemia; 41 cases of acute myeloid leukaemia (AML), 22 cases of chronic myeloid leukaemia (CML), and 26 cases of lymphoid leukaemia (LL). The groups proved to be separate entities (p < 0.000 25) and the differences between the groups were statistically significant (p < 0.001) except for the difference between AML and LL.

The 95% confidence limits for normal NAP scores were 15.0–132.6. Decreased scores were demonstrated in 73% of CML, 7% of AML but never in LL patients. Increased scores were found in 37% of AML, 31% of LL but never in CML patients. Evaluation of the distribution of the single cell NAP acivity (negative, weak positive, strong positive) showed decreased activity in 77% of CML, 15% of AML but never in LL patients. Increased activity was demonstrated in 63% of AML, 54% of LL and 9% of CML patients. The evaluation of single cell activity is a time‐saving method, which furthermore proved superior to the scoring method in discriminating between the types of leukaemia investigated.
Adaptation of the May‐Grünwald‐Giemsa staining method for automated differential counting of blood leucocytes by a Hematrak analyzer
Authors: Penttilä I.M. , Mahlamäki E. , Mononen I. , Kärkkäinen P.
Published: August 2010

The selection of exact pH (6.85/6.90) concentrations of the stains and staining times made it possible to use the May‐Grünwald‐Giemsa (MGG) staining method for smears studied with a Hematrak model 240 automated differential leucocyte counter. The differences between the smears stained by the method of Wright according to the recommendations of the manufacturer and the present MGG‐modification resulted in non‐significant differences in the recognition of various types of leucocytes.

The differences between the manual method and those obtained by Hematrak for the MGG‐stained samples with normal total leucocyte counts were nonsignificant with all types of leucocytes except basophils, which were underestimated by the instrument. The present staining procedure was adequate in studying of neutrophilias. When samples with lymphocytosis or with immature cells were examined, too few lymphocytes and immature cells and too many monocytes were obtained by the instrument, as compared to the manual method.
Drug‐indiced agranilocytosis: Evidence for the commitment of bone marrow haematopoiesis
Authors: Heit W. , Heimpel H. , Fischer A. , Frickhofen N.
Published: August 2010

Clinical and haematological features of 61 patients with drug‐induced agranulocytosis (63 episodes) are presented. Multiple drug consumption was a common observation and complicated the attempt to incriminate a particular drug as being aetiologically involved. Bone marrow analysis shortly after the diagnosis revealed evidence for an impairment of proliferative granulopoiesis in the majority of cases. This observation was confirmed by in vitro culturing of granuloid precursor cells (CFU‐c). Moreover, the data clearly demonstrated that drug‐induced agranulocytosis may not be restricted to the granulocytic series. Thrombocytosis and reticulocytosis during the recovery phase are taken as an indication for the commitment of all haemopoietic cell lineages in agranulocytosis. These observations were in accordance with cytomorphological studies and in vitro culture data of erythroid precursor cells (CFU‐e, BFU‐e) of bone marrow aspirates taken in the initial phase of agranulocytosis. More than 25% of the patients showed a marked erythroid depression in the marrow.
Leukaemia cell mobility in childhood acute myeloid leukaemia based on the FAB classification
Authors: Yamazaki Munehiro , Komiyama Atsushi , Yamazaki Toshio , Yasui Kozo , Morohashi Fumio , Miyagawa Yukiaki , Akabane Taro
Published: August 2010

Leukaemia cell mobility was evaluated by the agarose plate method in 20 children with acute myeloid leukaemia (AML) based on the FAB classification. M1 blasts from 3 patients did not show any migration, probably reflecting their cellular immaturity. Leukaemia cells from 3 of 8 patients with M2 AML showed random migration, but those from the remaining 5 did not have this ability. The results thus demonstrated the heterogeneity of M2 AML in the function as well as the morphology. The M3 leukaemia cells failed to migrate under agarose in 2 patients studied, implying that a certain cytoskeletal defect in the cells. In all of 2 M4, 2 M5A and 3 M5B AML patients, leukaemia cells showed active mobility. The random migration value (mean ± SD) of M5A and M5B cells was 5.7 ± 1.7 compared with that of normal monocytes of 4.0 ± 0.4 (p > 0.01), indicating the enhanced random migration of leukaemia monocytes. M4 and M5B leukaemia cells had chemotactic responsiveness to zyrnosan‐activated serum comparable to that of normal monocytes, but M5A cells were different from them in lacking apparent chemotactic responsiveness to the factor. Lymphadenopathy and leucocytosis were more prominent in the patients whose leukaemia cells were capable of migration than in those not having such cells. Mobility of leukaemia cells probably reflects their cell lineage, cellular maturity and functional distortion during leukaemic transformation, and appears to be related to clinical features.
Phase III study of intermittent 5‐drug regimen (VBCMP) versus intermittent 3‐drug regimen (VMP) versus intermittent melphalan and prednisone (MP) in myelomatosis
Authors: Hansen Ole Paaske , Clausen Niels‐Aage Tøffner , Drivsholm Aage , Laursen Benedicte
Published: August 2010

A prospective randomized trial in 96 patients with previously untreated myelomatosis was performed comparing 3 regimens of chemotherapy: (i) Intermittent vincristine, BCNU, cyclophosphamide, melphalan, and prednisone (VBCMP) to (ii) intermittent vincristine, melphalan and prednisone (VMP) to (iii) intermittent melphalan and prednisone (MP). Induction response rates and survival were similar in all 3 regimens. An improvement in relapse‐free survival was observed by adding vincristine to MP, but this did not achieve statistical difference (p = 0.10). Patients given VBCMP fared slightly worse than those given VMP. The haematologic toxicity was similar in all 3 regimens, but the tolerability of VBCMP was lower. Although showing no statistical differences between the 3 treatment regimens, the results support the view that a combination of MP ‘standard’ induction therapy in MM with frequently administered vincristine has a trend towards postponing treatment failure due to development of resistance to melphalan.
Immunocytochemical evidence of common‐ALL antigen in null‐ALL
Authors: Basso Giuseppe , Capuzzo Fabrizia , Simioni Ines , Destro Roberta , Gazzola Maria Vittoria , Cocito Maria Grazia , Cozzi Marzia , Milanesi Carla , Putti Maria Caterina , Carli Modesto , Zanesco Luigi
Published: August 2010

4 cases of acute lymphoblastic leukaemia (ALL), diagnosed as null‐ALL by indirect immunofluorescence using monoclonal antibodies, were similarly investigated using a sensitive immunoperoxidase method. The Avidin ‐ Biotin system was employed. The immunoenzymatic results were in agreement with those obtained with immunofluorescence techniques for all antigens except common‐ALL (C‐ALL). The C‐ALL antigen, recognized by the J5 antibody, was detected only by the immunoperoxidase method on cell membranes of the 4 ALL. This paper discusses the possibility of false negative results in testing for C‐ALL antigen by conventional indirect immunofluorescence as suggested by refined immunocytochemical screening. Moreover, the ability of the immunoperoxidase system to identify antigens on cell membranes, even at very low density, is discussed. The clinical significance of the presence of C‐ALL antigen at weak intensity in cases of null‐ALL is also considered.
Changes in plasma antithrombin (heparin cofactor activity) during intravenous heparin therapy: Observations in 198 patients with deep venous thrombosis
Authors: Holm H. A. , Kalvenes S. , Abildgaard U.
Published: August 2010

Plasma antithrombin (AT) measured as heparin cofactor activity decreased 0.16 ± 0.13 U/ml (mean ± SD) in 198 patients who received heparin infusion during 1 wk for deep venous thrombosis (DVT). The decrease was weakly, but significantly correlated to heparin dose (r = 0.15, p = 0.02) and to heparin plasma concentration (r = 0.12, p = 0.05). In patients with subnormal AT at start of heparin treatment, AT decreased less and tended to normalize at the end of heparin infusion, suggesting increased synthetic rate. The decrease in AT was apparently unrelated to the extension or the fate of the thrombus, and also unrelated to other patient and disease characteristics apart from a significantly higher decrease in diabetics. 5 out of 9 patients with AT values below 0.60 U/ml had serious disease, and 1 died from pulmonary embolism (PE) shortly after cessation of heparin (AT 0.22 U/ml). We conclude that the main decrease in AT occurs during the initial 3 d of heparin treatment. If AT stays above 0.70 U/ml after 3 d of treatment, further AT monitoring is hardly indicated.
Labelling of granulocytes with radioindium: Cell isolation and labelling parameters
Authors: Syrjälä Martti T.
Published: August 2010

A method for isolation and labelling of purified granulocytes with 111In‐oxine for scintigraphy of inflammatory processes is described. The procedure, on average, yielded 27.6% of the initial granulocytes in the sample with minimal lymphocyte and platelet contamination. The mean erythrocyte/granulocyte ratio was 0.95. In 20 experiments, 85.3% of the total radioactivity was attached to granulocytes, 8.7% to erythrocytes and 5.9% was in the plasma. The sensitivity and the specificity for the detection of acute inflammatory processes in 44 patients were 91.3% and 81.870, respectively.
Improvement of post platelet transfusion increments following intravenous immunoglobulin therapy for leukaemic HLA‐immunized patients
Authors: Atrah H. I. , Sheehan T. , Gribben J. , Crawford R. J. , O'Donnel J. R. , Sandilands G. P.
Published: August 2010

3 patients with acute leukaemia, HLA antibodies and thrombocytopenia refractory to random donor platelet transfusions were treated with high‐dose i.v. immunoglobulin. All 3 patients responded favourably with improved post‐transfusion recovery of random platelets. In 1 patient, the recovery of transfused histocompatible platelets was also enhanced. Treatment was followed by reduction in the total lymphocyte count and marked changes in lymphocyte subsets in 1 patient.
Bone marrow transplantation in multiple myeloma
Authors: Tura Sante , Cavo Michele , Baccarani Michele , Ricci Paolo , Gobbi Marco
Published: August 2010

We report 3 cases of allogeneic bone marrow transplantation in multiple myeloma that we have recently performed. Following conditioning treatment with i.v. cyclophos‐phamide (60 mg/kg/d, for 2 d), oral melphalan (1.0 mg/kg/d, for 5 d), i.v. BCNU (5.5 mg/kg, in a single dose) and total body irradiation (10 Gy in a single fraction) we observed in all 3 cases the disappearance both of serum M component and of monoclonal bone marrow plasma cells. 1 patient died of acute GVH disease, grade IV, at 2 months, while the other 2 patients are in good health and in unmaintained complete remission at 4 and 20 months, respectively. The usefulness of allogeneic bone marrow transplantation in the management of multiple myeloma is emphasized.