bluuebunny/crossref_metadata_2025 huggingface dataset:
Generation of non MHC restricted killing in non‐cytotoxic T‐lymphocytes from leukaemia patients
Authors: Matera Lina , Cardoso Estela , Caudana Luisella , Moore Michael
Published: March 2008

Summary.Experiments were undertaken to determine whether the depression of natural killer (NK) activity previously observed in the peripheral blood lymphocytes (PBL) of leukaemia patients in remission extended to NK‐like precursors in the blood. T‐lymphocytes (E +) from leukaemia patients and normal subjects were depleted of IgG Fc‐receptor‐bearing (YFcR) fresh NK cells by passage over immune complex‐coated monolayers.yFcR —E + PBL were cultured alone or with DAUDI cells. On day 5 of culture, cytotoxicity toward the NK‐sensitive cell lines K562 and MOLT‐4 was evaluated in the responder lymphocytes of leukaemia patients and controls. Negligible NK‐like cytotoxicity was found in both FcR —E +PBI, responder populations cultured alone. By contrast, stimulation with DAUDI induced high levels of K562 and MOLT‐4 cytotoxicity in leukaemia as well as in normal responder cells. Complement‐mediated cytotoxicity experiments using various McAb demonstrated that in both normal and leukaemia cultures NK‐like effectors react with the pan‐T OKT3 McAb and with the OKTl1 McAR directed to the SRBC receptor, but not with Leullb and OKMl McAbs, directed against antigens expressed on peripheral blood NK cells. Fractionation of the responder cells on discontinuous Percoll gradients showed that most of this activity was present in the highly dividing blast cell fraction.
Increased concentration of the fast‐acting plasminogen activator inhibitor in plasma associated with familial venous thrombosis
Authors: Jørgensen Maja , Bonnevie‐Nielsen Vagn
Published: March 2008

Summary. We have earlier demonstrated that in a family with a tendency to recurrent venous thrombosis the release of tissue plasminogen activator (t‐PA) activity in blood after stimulation was abnormally low. This observation could be related either to an impaired release of t‐PA into the blood stream or to a masking of the released t‐PA by a high concentration of PA inhibitor(s). In order to distinguish between these two possibilities the family was reinvestigated using various newer techniques, including an ELISA for t‐PA, an assay for quantitation of the fast‐acting PA inhibitor and SDS polyacrylamide gel electrophoresis followed by fibrin‐enzymography. Hereby the family members were demonstrated to have a high concentration in plasma of the PA inhibitor. After stimulation the release of t‐PA into the blood was normal, the t‐PA activity, however. was immediately inactivated by complex formation with the fast‐acting PA inhibitor.
The fate of intravenously administered hepatic ferritin in normal, phenylhydrazine‐treated and scorbutic guinea‐pigs
Authors: Simon Monica , MacPhail Patrick , Bothwell Thomas , Lyons Gillian , Baynes Roy , Torrance John
Published: March 2008

Summary. When highly purified hepatic 59Fe‐ferritin was injected intravenously into normal guinea‐pigs more than half of it was taken up by red cell precursors and the iron was used for haem formation. This was studied in more detail in animals in which a reticulocytosis had been induced either by phenylhydrazine or by repeated venesections. 55% of the injected ferritin iron was found in reticulocytes at 1 h. Experiments using ferritin doubly labelled with 59Fe and 125I indicated that the whole molecule was taken up, with two‐thirds of the radioactivity being associated with the membrane at 1 h and one third being already within the cell. There was a progressive loss of 125I activity over the ensuing hours, while most of the 59Fe was slowly internalized and incorporated into haem between 1 and 24 h. In contrast, 90% of the activity taken up by red cell precursors from 59Fe‐transferrin was present as haem at all times. The liver and spleen were the two other major sites of 59Fe‐ferritin uptake in phenylhydrazine treated animals. While there was an early uptake of 59Fe into haem in these organs, some redistribution occurred with time, since most of the 59Fe was in a non‐haem fraction by 24 h. In a final experiment the distribution and fate of 59Fe‐ferritin was studied in scorbutic animals treated with phenylhydrazine. The findings were similar to those in normals similarly treated, which suggests that ferritin iron was being effectively mobilized for haem formation despite the ascorbic acid depletion.
Diagnostic and prognostic value of monoclonal antibodies in immunophenotyping of angioimmunoblastic lymphadenopathy/lymphogranulomatosis X
Authors: Knecht Hans , Odermatt Bernhard F. , Maurer Robert , Rüttner Jacques R.
Published: March 2008

Summary. Immunophenotyping of frozen lymph node sections from seven patients with morphologically defined angioimmunoblastic lymphadenopathy (AILD) was performed with a panel of 20 monoclonal antibodies (MoAb). MoAb for identification of dendritic reticulum cells showed remnants of follicular structures in all cases. In four cases these ‘burnt out’ follicular structures were associated with clusters of polyclonal B‐cells. Vascular proliferation and small amounts of intercellular collagenous fibrils were reliably revealed by MoAb CIV22 against basement membrane collagen. Cellular infiltrates of T4 + cells prevailed in four cases, of T8+ cells in one case. In one case conventionally identified blasts were of T‐and B‐cell origin. Expression of cell proliferation associated nuclear antigen identified by MoAb Ki‐67 was present in more than 25% of lymph node cells in three patients with fatal outcome, but in less than 10% of cells in two patients with a better clinical course. Therefore MoAb Ki‐67 may represent a valuable prognostic marker in AILD.
Abnormalities of chromosome 12p 13 and malignant proliferation of eosinophils: a nonrandom association
Authors: Keene P. , Mendelow B. , Pinto M. R. , Bezwoda W. , MacDougall L. , Falkson G. , Ruff P. , Bernstein R.
Published: March 2008

Summary. Four patients representing a spectrum of haematological malignancies are reported. Two patients had Philadelphia chromosome negative myeloproliferative disorders, one had acute lymphoblastic leukaemia and one had eosinophilic leukaemia. In each case eosinophilia was present and demonstrated to be part of the malignancy by the association of clonally abnormal metaphases with eosinophil granules. Abnormalities involving the short arm of chromosome 12 (12p 13) were a constant feature in all four cases and therefore a nonrandom association between this chromosome region and malignant eosinophil proliferation is proposed.
Cell kinetic effect of low dose arabinosyl cytosine
Authors: Baccarani Michele , Tazzari Pier Luigi , Motta Maria Rosa , Rizzi Simonetta , Fanin Renato , Fasola Gianpiero , Damiani Daniela , Dinota Angelo , Tura Sante
Published: March 2008

Summary. Low dose arabinosyl cytosine (ARA‐C) is effective for treatment of acute non‐lymphocytic leukaemia (ANLL). The mechanism of action is not clearly understood and it was suggested that low doses of the drug could induce leukaemic cells to differentiate. We investigated the effect of low dose ARA‐C (20 mg/m2/d, divided into two doses s.c. at 12 h intervals, × 20 d) on the cell cycle distribution of leukaemic cells in four cases of ANLL. By comparison, four other cases of ANLL were studied during treatment with standard dose ARA‐C (200 mg/m2/d as a continuous i.v. infusion × 7 d). Both treatments induced an accumulation of leukaemic cells in post G1 phases, at a variable extent and rate. During treatment by low dose ARA‐C, the mitotic index (MI) fell slowly to zero in two patients who achieved a complete remission (CR), while it fell but recovered during treatment in the patients who did not achieve a CR. The MI fell rapidly to zero in the four cases treated by standard dose, who achieved a CR. These data are consistent with the known cytotoxic activity of ARA‐C, via inhibition and slowing of DNA synthesis leading to defective cell proliferation and to cell death.
Multiple myeloma: the relationship between CALLA (CD10) positive lymphocytes in the peripheral blood and light chain isotype suppression
Authors: Wearne Alaine J. , Joshua Douglas E. , Brown Ross D. , Kronenberg Harry
Published: March 2008

Summary. This study describes the presence of small numbers of common acute lymphocytic leukaemia antigen (CALLA, CD10)‐positive lymphocytes in the peripheral blood of patients with multiple myeloma. A significant correlation (0.001 < P <0.01) was found between the lack of light chain isotype suppression (LCIS), which is characteristic of progressive myeloma, and the presence of CALLA‐positive lymphocytes. Sixty patients with multiple myeloma, four with benign monoclonal gammopathy (BMG) and seven with solitary plasmacytoma (SP) were monitored in this study. Nineteen of the patients with multiple myeloma demonstrated LCIS, of which only three were found to have CALLA‐positive lymphocytes. Of the 41 patients with multiple myeloma who did not have LCIS, 20 (49%) had CALLA‐bearing lymphocytes. None of the patients with BMG or SP demonstrated LCIS or were found to have CALLA‐bearing lymphocytes in their blood. Forty‐four of the patients with multiple myeloma were also monitored for serum beta‐2‐microglobulin (SB2M levels. There was no correlation between the SB2M and either LCIS or CALLA‐positivity. Detection of CALLA‐positive lymphocytes in the blood of patients with multiple myeloma may be an early marker of the onset of progressive disease. The correlation of CALLA expression on lymphocytes with lack of LCIS provides further evidence for the operation of immunoregulatory systems in these patients.
Flow cytochemical patterns of white blood cells in human haematopoietic malignancies: II, CHRONIC LEUKAEMIAS
Authors: Drewinko B. , Bollinger P. , Brallas C. , Wyatt J. , Simson E. , Trujillo J. M.
Published: March 2008

Peripheral blood samples from 73 patients with chronic leukaemia were measured with the Technicon H‐6000 automated haematology analyser to provide flow cytochemical (peroxidase content) and volume (light scatter) discriminated scattergram patterns. For chronic granulocytic leukaemia (CGL). these patterns were so reproducible and distinct that they allowed an immediate diagnosis even without the benefit of microscopic examination, Relative and absolute basophilia was an invariable feature, and remained detected by the H‐6000 even when the patient was in haematologic and cytogenetic remission or progressed into blast crisis (BC). Meet patients in BC also demonstrated an inordinately high number oflarge unstained cells (LUC) and high proportions of ‘lymphocytes’(small blasts with no peroxidase content by visual inspection). Thus, for patients with CGL, LUC values above 10%, and/or steady increments in the proportion of ‘lymphocytes’, merit concern as these changes may herald an accelerated phase of disease.The scattergram pattern of untreated chronic lymphocytic leukaemia (CLL) showed a dense accumulation of data points within the lymphocytic ‘box’with a small cluster of granulocytic elements. Most patients also had a frankly abnormal proportion of LUC. Sixteen patients with CLL were compared for ratios of LUC to lymphocytes and stage of disease; patients with the most advanced stage (IV) had the highest, statistically significant values, than the patients with more benign disease. Thus, it is possible that follow up with this instrument of patients with CLL will also allow early detection of an impending prolymphocytoid transformation (accelerated phase) of this disease.
Reduced risk of non‐A, non‐B hepatitis after a first exposure to ‘wet heated’factor VIII concentrate
Authors: Kernoff P. B. A. , Miller E. J. , Savidge G. F. , Machine S. J. , Dewar M. S. , Preston F. E.
Published: March 2008

The risk of post‐infusion non‐A, non‐B hepatitis (NANBH) in patients receiving a first exposure to unheated or conventionally ‘dry heated’factor VIII concentrates approaches 100%. implying invariable contamination of these products. Amongst 18 patients who received a first treatment with a ‘wet heated’commercial concentrate, five (28%) developed asymptomatic NANBH, suggesting a more efficient inactivation of NANB agent(s) by this process. 2/9 (22%) of the batches of concentrate used in the study were implicated in NANBH transmission. One of these two batches. responsible for NANBH in four patients, had been prepared from a plasma pool containing an unusually large proportion of donations with high alanine aminotransferase (ALT) levels. A resulting high level of viral contamination in this batch may have been sufficient to override the effects of the sterilization process. All patients remained anti‐HIV seronegative at 17–28 months of follow‐up.
Clotting factor levels and the risk of diffuse microvascular bleeding in the massively transfused patient
Authors: Ciavarella D. , Reed R. L. , Counts R. B. , Baron L. , Pavlin E. , Heimbach D. M. , Carrico C. J.
Published: March 2008

Clotting factor activities and coagulation screening tests in 36 massively transfused patients were measured after every 12 units of blood and whenever diffuse microvascular bleeding (MVB) developed. Moderate deficiencies in clotting factors were common, but they were not associated with MVB. MVB was associated with severe abnormalities of coagulation, i.e. a fibrinogen level <0.5 g/l or clotting factor levels <20%.

The quantitative relationship between the prothrombin (PT) and partial thromboplastin (PTT) times and underlying clotting factor levels was explored by multiple linear regression. Clotting factor levels accounted for only 65–85% of the variability in these tests. However, clotting factor activities less than 20% were reliably reflected by marked prolongations of the PT and PTT (values greater than 1.8 times control).

Our data suggest that commonly used replacement formulas are not likely to prevent MVB, since consumption of platelets and/or clotting factors, rather than simple dilution, is a major cause of the deficiencies leading to MVB. Modified whole blood alone was sufficient replacement therapy for most patients. Guidelines for transfusion of supplemental components during massive transfusion are given.
Abnormal calcium transport into microsomes of grey platelet syndrome
Authors: Enouf Jocelyne , Lebret Maryline , Bredoux Raymonde , Levy‐Toledano Sylviane , Caen Jacques P.
Published: March 2008

Summary. Calcium uptake into isolated membrane vesicles from two patients with a grey platelet syndrome has been investigated. An increase in calcium transport appears in both patients when compared to controls. Determination of the kinetic parameters of the calcium transport system gave similar apparent affinity for calcium and an increase in the calcium uptake velocity. This increase in calcium transport is correlated with the increase of the associated Ca2+ activated ATPase activity. The results would suggest a new relationship between the ultrastructural and functional abnormalities of the grey platelet syndrome.
Haemopoietic reconstitution after allogeneic bone marrow transplantation in man: recovery of haemopoietic progenitors (CFU‐Mix, BFU‐E and CFU‐GM)
Authors: Ma David D. F. , Varga Dianne E. , Biggs James C.
Published: March 2008

Haemopoietic reconstitution was evaluated in 44 patients given HLA compatible sibling bone marrow transplants. The mean peripheral blood haemoglobin, neutrophil and platelet counts were markedly reduced early post‐graft but returned to normal by 26 weeks post transplant. Bone marrow multipotent (CFU‐Mix), erythroid (BFU‐E) and myeloid (CFU‐GM) progenitor cell reconstitution were also assessed at regular intervals up to 2 years post‐graft. The mean value of CFU‐GM increased gradually and attained a normal value by 52 weeks. The BFU‐E value did not reach a normal value until after 52 weeks post‐graft. However, CFU‐Mix growth appeared to be impaired even up to 2 years post‐transplant. The occurrence of graft versus host disease at 3 months post‐transplant was associated with significantly lower mean numbers of platelets, marrow CFU‐GM, BFU‐E and CFU‐Mix. Post‐transplant patients who were on methotrexate therapy were also shown to have lower marrow CFU‐GM and neutrophil values compared to those patients who received cyclosporin post‐transplant. This study demonstrated that although peripheral blood counts were normal after 26 weeks post‐graft, marrow stem cell reserve in these patients was reduced. This might in part explain the documented increase in risk of severe infections or thrombocytopenia in some of these patients, particularly during viral infection, graft‐versus‐host disease or immunosuppressive treatment.
Rearrangement for the T‐cell receptor gene and co‐expression of immature T‐cell markers and natural killer cell phenotype, in a patient with acute lymphoblastic leukaemia
Authors: Pizzolo G. , Trentin L. , Vinante F. , Agostini C. , Zambello R. , Ranucci A. , Luca M. , Chilosi M. , Dazzi F. , Foa R. , Caligaris‐Cappio F. , Perona G. , Semenzato G.
Published: March 2008

We describe a patient with acute lymphoblastic leukaemia whose blasts co‐expressed immature T‐cell markers and nearly the entire phenotypic repertoire of NK cells. The T‐cell nature of the proliferating blasts was proven by the demonstration of the rearrangement for the β‐chain of the T‐cell antigen receptor. Although an abnormal phenotypic expression related to the neoplastic proliferation cannot be formally excluded, it is possible that the cells in this patient may represent the clonal expansion of a normal subpopulation of T‐cell lineage NK‐related cells frozen at an early stage of differentiation. These features provide arguments for discussing the controversial issue of the ontogeny of NK cells and their relationship to the T‐cell lineage.
The relationship between chronic lymphocytic leukaemia and prolymphocytic leukaemia
Authors: Melo Junia V. , Catovsky Daniel , Gregory Walter M. , Galton David A. G.
Published: March 2008

The prognostic value of biological, clinical and laboratory features was analysed in a series of 265 patients with chronic lymphocytic leukaemia (CLL) and prolymphocytic leukaemia (PLL). On univariate analysis seven features were shown to influence significantly the survival of the whole group of patients: absolute prolymphocyte count (ABS PROL), percentage of prolymphocytes (%PROL), WBC, spleen size, age, intensity of surface‐membrane immunoglobulin (SmIg) and mouse (M) rosettes. Multivariate regression analysis of these features showed that only ABS PROL and spleen size had independent prognostic significance. The survival in PLL (38 cases) was significantly shorter than in CLL (227 cases) (median survival=3 and 8 years, respectively). Patients with CLL with an increased %PROL (11–55%), defined as CLL/PL, could be divided into two groups: those with ABS PROL ≤ 15 × 109/1 (26 cases) fell within the ‘standard‐prognostic risk’ for typical CLL (i.e. ≤ 10% PROL), whereas the survival outlook for the cases with ABS PROL > 15 × 109/1 (40 cases) was as bad as for PLL. A scoring system was generated with the four features that showed high prognostic significance: ABS PROL, spleen size, SmIg and M‐rosettes. The score proved to be superior to any single feature as a predictor of survival, being especially useful in the analysis of the CLL/PL group: cases with high scores (>2) had a median survival of 2·5 years, while the median has not been reached for those with low scores (≤2). We suggest that this scoring system may help to identify the cases of CLL/PL that behave as PLL, and as such may benefit from different treatment.
The spleen and haemolysis: evaluation of the intrasplenic transit time
Authors: Ferrant A. , Leners N. , Michaux J. L. , Verwilghen R. L. , Sokal G.
Published: March 2008

The mean intrasplenic red cell transit time (STT) and the slow mixing splenic red cell volume (SSV) have been measured in patients with hereditary spherocytosis (HS), autoimmune haemolytic anaemia (AIHA) and lymphoproliferative disease (LD). There was an inverse relationship between the mean red cell life span (MRCLS) and the STT in HS (r=–0·96, P<0·001) and in AIHA (r=–0·90, P<0·001). No such relationship existed in LD. The size of the spleen and the SSV were not related to the severity of haemolysis. Our data offer strong evidence for the conditioning effect of the spleen on HS‐ and AIHA red cells and suggest that the STT is an index of the adverse effect of the spleen on red cells in patients with HS or AIHA.
Chromatographic analysis of human erythrocyte pyrimidine 5′‐nucleotidase from five patients with pyrimidine 5′‐nucleotidase deficiency
Authors: Hirono Akira , Fujii Hisaichi , Natori Hideyo , Kurokawa Izumi , Miwa Shiro
Published: March 2008

Human erythrocyte pyrimidine 5′‐nucleotidase (P5N) was separated into two subclasses, P5N‐I and P5N‐II, by DEAE Bio‐Gel A column chromatography. Their enzymological properties were studied using five normal subjects and five patients with different P5N deficiencies. Study of the normal subjects showed that P5N‐I and P5N‐II have distinctive properties, and P5N‐II is similar to the 5′‐nucleotidase in rat liver cytosol. The P5N‐II from the five subjects with this deficiency had normal activity and other normal enzymological properties. However, the P5N‐I from these patients had abnormal properties, including reduced activity. These variant enzymes had a high Michaelis constant for substrate cytidine 5′‐monophosphate and were heat stable. The optimum pH was shifted towards the acidic side in two patients, towards the basic side in one, and was unchanged in the other two. These results strongly suggest that the main cause of P5N deficiency is an abnormality of P5N‐I, probably arising from a structural gene mutation.
A preparation for calibrating the assay of the blood group antibody anti‐c
Authors: Phillips P. K.
Published: March 2008

A freeze‐dried, working calibration preparation (84/628) has been assayed against the International Incomplete anti‐c Reference Serum (67/160) in a multi‐centre study. The use of 84/628 will avoid the between‐laboratory variation, inherent in the use of local preparations, in the reporting of assays of anti‐c using the AutoAnalyzer. A value of 13 IU/ampoule has been assigned to the anti‐c content of preparation 84/628 which is available from the Blood Group Reference Laboratory, Oxford.
Demonstration of abnormal factor VIII multimers in acquired von Willebrand's disease associated with a circulating inhibitor
Authors: Ball J. , Malia R. G. , Greaves M. , Preston F. E.
Published: March 2008

We have studied the factor VIII multimeric structure in four patients with acquired von Willebrand's disease associated with a circulating inhibitor to the factor VIII complex. In three of the four patients tested, the high and medium molecular weight bands were absent when assessed by sodium dodecyl sulphate–agarose electrophoresis. Plasma from the fourth patient contained all the factor VIII multimeric forms, although the high molecular weight bands were markedly decreased in concentration. Intravenous infusion of 1‐deamino‐(8‐d‐arginine)‐vasopressin (DDAVP) resulted in the appearance of a full complement of multimers in the plasma of the two patients tested. This response was, however, transient and a return to pre‐infusion multimeric composition occurred within 2–4 h.

In the patients studied, an inhibitor to the factor VIII complex has induced an acquired variant von Willebrand's disease which transiently corrects after the infusion of DDAVP.
Delineation of receptor‐mediated colony‐stimulating factor (CSF‐1) utilization and cell production by precursors of mononuclear phagocytic series at various stages of differentiation
Authors: Chen Ben D.‐M. , Chou Ta‐hsu , Clark Carl R.
Published: March 2008

Summary. Colony‐stimulating factor‐1 (CSF‐1) is a specific haematopoietic growth factor that stimulates the production of macrophages by both bone marrow macrophage precursors (GM‐CFC) and certain more mature peripheral tissue macrophages. The relationship of CSF‐1 utilization and cell production by macrophage precursors at various stages of differentiation was studied. Bone marrow GM‐CFC had the highest proliferative capacity followed by blood monocytes and peritoneal exudate macrophages (PEM) as determined by their cell doubling time (DT) which was also dependent on the concentrations of exogenous CSF‐1. PEM had the longest initial lag period before commencing cell proliferation. Exogenous CSF‐1 was constantly utilized by the growing cells; depletion of available CSF‐1 resulted in growth arrest and, subsequently, cell death. The production of macrophage progeny, per amount of CSF‐1, correlated with parent macrophage maturity; for each 100 U of CSF‐1 consumed, bone marrow precursor cells and blood monocytes were capable of producing 17.9 × 104 and 13.4×104 progeny, respectively, whereas PEM generated only 4.6×104 daughter cells. Thus, the removal and destruction of CSF‐1 by more mature, less proliferative tissue macrophages may provide a possible mechanism by which CSF‐1 levels are reduced and the production of early haemopoietic macrophage precursors controlled.
Erythropoietic colonies in a serum‐free system: results in primary proliferative polycythaemia and thrombocythaemia
Authors: Eridani S. , Dudley J. M. , Sawyer B. M. , Pearson T. C.
Published: March 2008

Summary. The formation of erythropoietic colonies from the peripheral blood of normal subjects, patients with primary proliferative polycythaemia (PPP) and primary thrombocythaemia (PT) was studied, using a chemically defined serum‐free (S ‐) medium. Colony formation was markedly more prominent in the presence of burst‐promoting activity (BPA) and erythropoietin (Ep) than with medium alone (P<0.001). In cultures using medium alone, significantly more PPP patients formed colonies than the control group (P<0.05). In the PT group this difference did not achieve statistical significance, but the mean BFU‐E yield was significantly greater than in controls (P<0.05). In a separate series of experiments, parallel cultures in serum‐containing (S +) and serum‐free (S ‐) systems, in the presence of BPA and Ep did not show any significant difference in colony yield. The growth of‘endogenous’colonies in cultures with serum‐free medium alone could be due to a peculiar sensitivity of erythropoietic progenitors to growth factors other than Ep.
Determination of factor IX allotypes for carrier identification in haemophilia B
Authors: Wallmark A. , Ljung R. , Nilsson I. M.
Published: March 2008

Summary. The existence of two genetic variants (allotypes) of normal human factor IX is used for carrier detection in three families with severe and one with mild haemophilia B. By analysis of IX: Ag with two different monoclonal antibodies in 93 members of the families, allelic assignment is shown to be a complement in carrier diagnosis to genotypic DNA studies. Allelic assignment makes possible a reliable diagnosis based on phenotypic studies, though its usefulness is limited due to ethnic variation in allelic frequency. Determination of factor IX allotypes should be useful for carrier detection in many Swedish families with haemophilia B.
Changes in the distribution and organization of platelet actin induced by diamide and its functional consequences
Authors: Spangenberg P. , Till U. , Gschmeissner S. , Crawford N.
Published: March 2008

Summary. Exposure of blood platelets to diamide (azodicar‐boxylic acid‐bis‐dimethylamide) results in oxidation of sulphydryl groups present in the cytoskeleton and other proteins. This results in dramatic changes in functional behaviour of the cells.

The distribution and level of organization of the major cytoskeletal protein actin has been studied analytically by the DNase‐I inhibition assay and morphologically by electron microscopy (EM) of Triton X‐100 treated platelets adherent to EM grids. Exposure to diamide results in a redistribution of actin within the cell reflected in an increase in cytoskeletal F‐actin and a concomitant decrease in cytosolic actin. The magnitude of these changes depends upon the concentration of diamide and the time of exposure. Diamide also alters platelet aggregatory functions in response to certain stimuli.

Treatment of normal human platelets with 0.1 mM diamide proceeds via disaggregation (5 min exposure to diamide), inhibition of aggregation (30 min exposure), to finally a normalization of the aggregation response after 60–120 min incubation with diamide. In parallel with the return to full functional response the distribution of F‐actin between the cytoskeleton and cytoplasmic compartments returns to the control pattern. Incubation of the platelets with 0.5 mM diamide for 60 or more minutes leads to total inhibition of the aggregatory ability. In these cells the cytoskeleton associated F‐actin remains significantly elevated and the structural organization of the cytoskeleton is markedly altered. In contrast to the network of filaments subadjacent to the surface membrane seen in unstimulated platelets, the cytoskeleton now shows electron dense zones in the more central parts of the cytoplasm. This diamide‐induced structural reorganization of platelet cytoskeletal elements, associated with the inhibition of functional responses, emphasizes the dynamic nature of the membrane‐cytoskeletal axis and its importance in the expression of shape changes and aggregatory phenomena in response to surface stimuli.
Familial myelodysplasia: progressive disease associated with emergence of monosomy 7
Authors: Paul B. , Reid M. M. , Davison E. V. , Abela M. , Hamilton P. J.
Published: March 2008

. Two brothers developed hypoplastic anaemia with the development in one of refractory anaemia with excess blasts (RAEB) accompanied by emergence of monosomy 7. Both brothers have a constitutional inversion of chromosome 1. Neither shows the increased chromosomal fragility of Fanconi's anaemia or its variants. This family is the third reported in which monosomy 7 has been found when leukaemic or preleukaemic transformation has occurred in patients with familial hypoplastic anaemia.
Bone growth and haemopoiesis: steroid reversible anaemia, myelofibrosis and increased bone formation in a child
Authors: Macdougall Lorna G. , Pettifor John M. , Patel Jayvant M.
Published: March 2008

Factors regulating the interaction between bone marrow haemopoietic cells, stromal elements and bone growth are poorly understood. Disturbance in the equilibrium between these elements can occur as the result of metabolic bone disease, haematologic disorders, neoplasia and infections. The present report concerns a child with myelofibrosis, hypoplastic/dyserythropoietic anaemia, osteo‐blast proliferation and increased bone formation. A positive tuberculin skin test and elevated EB virus titre indicated previous exposure toMycobacterium tuberculosisand Epstein‐Barr virus. No active focus of infection was identified and no improvement occurred following anti‐tuberculous therapy. A dramatic improvement occurred on corticosteroid therapy. Reticulocytosis was followed by an increase in haemoglobin and platelets and a decrease in ESR. Bone marrow fibrosis resolved and the marrow was repopulated with normal haemopoietic tissue. The bone abnormalities improved both radiologically and histomorphometrically. Relapse occurred when steroids were discontinued. Bone marrow tissue culture supernate from the patient during the active phase of the disease inhibited colony formation by normal marrow mono‐nuclear cells. This was reversed by steroid therapy. It is postulated that EB virus may have triggered osteoblast proliferation with resultant bony and haematologic changes. Response to corticosteroids could be explained on the basis of suppression of osteoblast activity and correction of fibroblast mediated suppression of haemopoiesis.
Stromal stem cells (CFU‐f) in yolk sac, liver, spleen and bone marrow of pre‐ and postnatal mice
Authors: Van Den Heuvel Rosette L. , Versele Sigrid R. M. , Schoeters Greet E. R. , J. Vanderborght Oscar L.
Published: March 2008

Our results demonstrate that in yolk sac, liver, spleen and femoral bone marrow of mice at ages ranging between 11 d of gestation and adult life, important changes in the stromal stem cell population (CFU‐f assay) occur which are correlated with haemopoiesis. (1) In the liver, spleen and bone marrow, high numbers of CFU‐f precede high haemo‐poietic stem cell values. (2) As haemopoiesis starts in the spleen. CFU‐f numbers in fetal liver are low. Similarly, CFU‐f numbers decrease in the spleen as bone marrow haemopoiesis starts. This suggests the existence of a migration stream of stromal stem cells. (3) In spleen and bone marrow, CFU‐f numbers decrease towards adult life as these organs maintain a stable haemopoietic activity.
Flow cytochemical patterns of white blood cells in human haematopoietic malignancies
Authors: Drewinko B. , Bollinger P. , Brailas C. , Moyle S. , Wyatt J. , Simson E. , Johnston D. , Trujillo J. M.
Published: March 2008

Peripheral blood samples from 118 patients with acute leukaemia (68 untreated; 50 treated) were measured with the Technicon H‐6000 automated haematology analyser. This instrument provides, in addition to measurements of the classical haematology parameters (i.e. cell counts, haemoglobin concentration, etc.), a differential count on 104 WBC effected by means of flow cytochemistry (peroxidase content) and volume (light scatter) discrimination. Disregarding RBC and platelet counts and their volume distribution profiles, the most important diagnostic parameters for leukaemic disease were the WBC count, the WBC differential count, and the proportions of large unstained cells (LUC) and high peroxidase (HPX) cells obtained by the automated differential count as well as the mean value of the WBC peroxidase content distribution (MPA). Granulocytic leukaemias had lower MPA than normal and lymphocytic leukaemias had MPA values above normal. M1 leukaemias were also characterized by large proportions of LUC and low fractions of HPX, while M2 leukaemias showed low LUC with high HPX. M3 leukaemias had low LUC and very high HPX. M4 leukaemias had large LUC and‘monocytic’components and a modest fraction of HPX. M5 leukaemias had very large numbers of LUC,‘monocytes’and‘lymphocytes’and a normal HPX. For M1 leukaemia, the presence of less than 7% LUC following induction treatment was related to morphological changes of normal cells induced by chemotherapy while LUC above 10% usually indicated unsuccessful induction associated with the presence of residual blasts. If treatment was successful, M2 and M3 leukaemias characteristically decreased their HPX population. All M4 leukaemias studied by us failed to enter remission and continued to display high proportions of HPX and LUC. Similarly, most M5 leukaemias had a poor response to treatment and always showed a very high proportion of LUC. Untreated lymphocytic leukaemias demonstrated high LUC, normal HPX and a high proportion of‘lymphocytes’. Hairy cell leukaemias showed almost equal proportions of‘lymphocytes’and LUC. Successful chemotherapy of all lymphoid leukaemia entities was associated with rapid decreases in LUC. slower decrements of‘lymphocytes’and moderate and transient increments in HPX. Thus, flow cytochemistry can assist not only in the segregation of acute leukaemias along with FAB classification with nonmorphologic criteria, but also in the follow up of patients with these diseases.
Pattern of fibrinolytic parameters in patients with gastrointestinal carcinomas
Authors: Kirchheimer Johannes C. , Huber Kurt , Wagner Oswald , Binder Bernd R.
Published: March 2008

Changes in the plasma levels of components of the fibrinolytic system have been investigated in 80 patients suffering from gastrointestinal carcinomas. Urokinase antigen (RIA), tissue‐type plasminogen activator antigen (ELISA) and plasminogen activator inhibitor (functional assay) were determined. patients with pancreatic and colorectal carcinoma and metastases as well as those without metastases revealed significantly increased plasma urokinase levels. Those with gall bladder or gastric carcinoma did not show significantly elevated urokinase antigen levels compared to age‐matched controls. Determination of tissue‐type plasminogen activator antigen in all four carcinoma groups did not reveal significant differences when compared to an age‐matched healthy control group. The concentrations of plasminogen activator inhibitor were significantly increased in all carcinoma groups: there being no differences between the patient groups with or without metastases. No correlations between the different parameters of the fibrinolytic system could be obained.
Pyrimidine nucleoside monophosphate kinase hyperactivity in hereditary erythrocyte pyrimidine 5′‐nucleotidase deficiency
Authors: Lachant Neil A. , Zerez Charles R. , Tanaka Kouichi R.
Published: March 2008

The pyrimidine nucleoside triphosphates (CTP, UTP) increase in the pyrimidine 5′‐nucleotidase (P5N) deficient red blood cell (RBC) to a greater degree than do the pyrimidine nucleoside monophosphates (CMP, UMP). Pyrimidine nucleoside monophosphate (PNMP) kinase phosphorylates CMP and UMP to their respective phosphodiesters. We tested the hypothesis that increased PNMP kinase activity contributes to the disproportionate increase in CTP and UTP in the P5N deficient RBC. CMP and UMP kinase activities were increased in high reticulocyte (4.4±2.1 and 8.5±3.3 μmol/ml RBC per minute) compared to normal RBC (2.8±1.0 and 6.0±2.5 μmol/ml RBC per minute). P5N deficient RBC (n= 2) had significantly increased CMP and UMP kinase activities (14.0 and 26.5 ±mol/ml RBC per minute). UMP and CDP‐ethanolamine were able to increase the activity of CMP kinase in crude haemolysate and the activity of partially purified enzyme. Since the Km for CMP of CMP kinase was 33 μmol/1 in P5N deficient RBC and since the CMP concentration is 25‐90 μmol/1 in the P5N deficient RBC, the enzyme should be nearly saturated with CMP in the P5N deficient RBC. Thus, PNMP kinase hyperactivity appears to contribute to the disproportionate increase in CTP and UTP in the P5N deficient RBC.
Splenic haematocrit and the splenic plasma pool
Authors: Zhang B. , Lewis S. M.
Published: March 2008

A method was established for estimation of plasma volume and splenic plasma pool by quantitative scanning using 113mIn labelled transferrin (TF). The method was used in 12 patients with various haematological disorders and degrees of splenomegaly. 113mIn‐TF consistently gave about 6% over‐estimation of plasma volume by comparison with the standard 125I HSA method. The splenic plasma pool ranged from 1‐2% to 11.4% of the total plasma volume. By concurrent measurement of splenic red cell pool the splenic haematocrit (SHct) was obtained: mean 0.51. SD 0.08: the SHct/PCV ratio was 1.21 (SD 0.31) and the SHct/body Hct ratio was 1.30 (SD 0.30). SHct was independent of PCV and body Hct but there was a trend to a lower SHct in cases where splenomegaly was more marked. Direct measurement of splenic plasma pool may help to elucidate the cause of increased total plasma volume in such patients.
β2‐Microglobulin, lysozyme and lactoferrin in cerebrospinal fluid in patients with lymphoma or leukaemia: relationship to CNS involvement and the effect of prophylactic intrathecal treatment with methotrexate
Authors: ÖBerg G. , Haullgren R. , Venge P.
Published: March 2008

Central nervous system (CNS) involvement in patients with leukaemia or lymphoma presents a diagnostic problem. This study was conducted to test whether combined measurements of various cellular markers such as β2‐microglobulin (β2m), lactoferrin (LF) and lysozyme (LYS) in the cerebrospinal fluid (CSF) might aid in the diagnosis of CNS involvement in such patients. Forty‐two patients were studied. Sixteen were considered to have CNS involvement and 26 showed no signs of such involvement. In the group with symptoms or signs of CNS involvement, nine patients out of 12 had increased total protein in CSF, 14 of 14 increased β2m, 14 of 16 increased LYS and five of 15 increased LF. In patients without CNS involvement total protein was increased in four of 25. β2min three of 21. LYS in four of 28 and LF in one of 28 patients. The differences were statistically significant (P<0.01, P<0.001, P<0.001 and P<0.05, respectively). Prophylactic intrathecal methotrexate treatment in patients with acute lymphoblastic leukaemia caused an increase in the CSF of β2m, LYS and LF but not of total protein, which may reflect a drug‐induced inflammatory reaction in the CNS.

We conclude that combined measurements of the three cell markers add to our understanding of the cellular reaction to malignant cells in the CNS in leukaemia and lymphoma and may be valuable supplements in the diagnosis of this CNS involvement.
Incidence and mechanism of neutropenia and thrombocytopenia in patients with human immunodeficiency virus infection
Authors: Murphy M. F. , Metcalfe P. , Waters A. H. , Carne C. A. , Weller I. V. D. , Linch D. C. , Smith A.
Published: March 2008

The incidence of lymphopenia, thrombocytopenia and neutropenia was studied in 105 homosexual men with HIV infection. Lymphopenia was common in patients with AIDS (75%), but its incidence in PGL (24%) was not significantly different from that in asymptomatic anti‐HIV positive (15%) homosexual men. Neutropenia and thrombocytopenia were found in patients with AIDS or PGL. but not in asymptomatic anti‐HIV positive homosexuals. The study suggests that the neutropenia and thombocytopenia in these patients were due to autoimmune destruction of neutrophils and platelets.
Monoclonal antibodies to factor VIII: their application in immunoblotting for the visualization of factor VIII in therapeutic concentrates and plasma
Authors: Furlong R. A. , Welch A. N. , Peake I. R.
Published: March 2008

Two monoclonal antibodies (McAb) termed 12A4 and 19C1 have been raised against human factor VIII. In immunoassays 12A4 bound to factor VIII antigen (VIII:Ag) in plasma but not serum whilst 19C1 bound to VIII:Ag in both plasma and serum. Both McAb were shown by immunoblotting to react with the carboxy (C) terminal polypeptide of factor VIII which appeared as a doublet with a molecular weight (Mr) of 77000/75000.

The C terminal factor VIII polypeptide was detectable by immunoblotting in each of 12 therapeutic factor VIII concentrates, from six different manufacturers, although its level was variable.

Factor VIII was visualized in plasma by immunoblotting following its immunoadsorption and elution from agarose‐bound monoclonal antibodies. No Mr 77000/75000 bands were detectable in plasma obtained from 13 unrelated CRM‐ haemophiliacs whilst 11 CRM+ haemophilic plasmas from seven kindred were shown to have a C terminal factor VIII polypeptide of normal molecular size.
The calcium permeability of cold‐stored erythrocytes
Authors: McNamara M. K. , Wiley J. S.
Published: March 2008

The calcium permeability of erythrocytes was studied by a new technique in which cells were loaded with an intracellular calcium chelator, quin 2, followed by measurement of 45Ca influx. The erythrocyte calcium permeability did not change during cold storage of blood in citrate‐phosphate‐dextrose for 6 weeks. Previous reports of an increased Ca2+ permeability in cold‐stored erythrocytes have been based on an ATP‐depletion technique which yields values for Ca2+ influx which are influenced by erythrocyte 2.3DPG levels.
3′‐Azido‐3′‐deoxythymidine (AZT) inhibits proliferation in vitro of human haematopoietic progenitor cells
Authors: Dainiak Nicholas , Worthington Michael , Riordan Mary Alice , Kreczko Sandra , Goldman Lyle
Published: March 2008

Peripheral blood cytopenias are a serious, dose‐limiting toxicity of AZT therapy in patients infected by HIV. To evaluate the mechanism by which cytopenias develop, AZT effects of haematopoietic differentiation and growth were measured in serum‐free, nucleoside‐depleted cultures of normal human bone marrow. In contrast to native thymidine, AZT suppressed the proliferation of erythroid, granulocyte/macrophage and primitive haematopoietic stem cells in a dose‐related and time‐dependent fashion. Relative progenitor sensitivity varied, with half‐maximal concentrations of 1–5 μM and 20–40 μM AZT for inhibition of erythroid and nonerythroid progenitor cell growth, respectively. Inhibition was observed over full ranges of concentrations of haematopoietic tissue‐specific regulators (human erythropoietin, colony‐stimulating activity, interleukin‐3 and lymphocyte conditioned medium) and of platelet‐derived growth factor (PDGF), an agent that enhances erythropoiesis in vitro via accessory marrow stromal elements. Furthermore, suppression was similar in cultures of marrow cells that were depleted of accessory populations, suggesting that its action is directed at progenitors. Finally, when deoxythymidine was added in increasing amounts to cultures with a half‐maximal concentration of AZT, inhibition was abrogated. We conclude that AZT is a potent inhibitor of haematopoiesis in vitro, and that erythroid progenitors are particularly sensitive to its action. These results may explain the marrow hypoplasia that occurs during AZT administration in vivo.
Immunocytochemical evaluation of the percentage of proliferating cells in pathological bone marrow and peripheral blood samples with the Ki‐67 and anti‐bromo‐deoxyuridine monoclonal antibodies
Authors: Falini B. , Canino S. , Sacchi S. , Ciani C. , Martelli M. F. , Gerdes J. , Stein H. , Pileri S. , Gobbi M. , Fagioli M. , Minelli O. , Flenghi L.
Published: March 2008

The monoclonal antibody Ki‐67, directed against a nuclear antigen expressed by dividing cells in all the phases of cell cycle except G0 and early G1, was used in combination with an anti‐BrdU monoclonal antibody, reacting selectively with cells in S‐phase, for assessing the percentage of proliferating cells in bone marrow and peripheral blood samples from patients with lymphoma, leukaemia and multiple myeloma. Immunocytochemical labelling of proliferating cells was performed on marrow frozen sections and/or cytospins using an immunoalkaline phosphatase (APAAP) technique that made it possible to obtain proliferative index measurements in a few hours in contrast to the 3–7 d needed with tritiated thymidine. In the 54 marrow lymphoma cases studied a highly significant correlation was observed between the proportion of Ki‐67 (+) cells and the separation into low‐ and high‐grade malignant lymphomas according to the Kiel classification. In patients with multiple myeloma at the first diagnosis, the percentage of Ki‐67 (+) cells was low (6–10%). In contrast, a high percentage of Ki‐67 (+) cells (40–50%) was observed in a young adult with multiple myeloma, in a patient who first presented at the clinical observation with an extradural mass and in three patients who developed extramedullary masses several years after the initial diagnosis of myeloma. In acute lymphoblastic leukaemias of common type the mean value of Ki‐67 labelling was 31.3%. Because of their simplicity and rapidity, immunocytochemical techniques may be expected to replace autoradiography and flow cytometry for the detection of proliferating cells in haematological samples.