bluuebunny/crossref_metadata_2025 huggingface dataset:
Adenosine Diphosphate Induced Platelet Aggregation and Release Reaction in Heparinized Platelet Rich Plasma and the Influence of Added Citrate
Authors: Heptinstall Stanley , Mulley Graham P.
Published: March 2008

Summary. Platelets in heparinized platelet‐rich plasma (PRP) from 54 volunteers were examined for their ability to aggregate and to release tritiated 5‐hydroxy tryptamine (3H‐5HT) in response to adenosine diphosphate (ADP). Release of 3H‐5HT was considerable in 26 of the preparations. When release was high, lower concentrations of ADP were required to induce irreversible aggregation. Second phase aggregation reflected extensive release reaction. Citrate always augmented the release induced by ADP.
In Vitro Thrombogenicity Tests of Factor IX Concentrates
Authors: Pepper D. S. , Banhegyi D. , Howie Ann , Cash J. D.
Published: March 2008

Summary. Three different types of test were compared in characterizing the properties of thrombogenic material in factor IX concentrates. The thrombin generation time (TGt50) of Sas et al (1975), the non‐activated partial thromboplastin time (NAPTT) of Kingdon et al (1975) and the factor VIII inhibitor bypassing activity (FEIBA) of Elsinger (1975) were used. When concentrates were incubated with purified antithrombin III (At III), the NAPTT lengthened but no effect was seen with the TGt50 and FEIBA. The TGt50 showed thermal sensitivity both in the presence and absence of antithrombin III. When concentrates were treated with the serine protease inhibitor diisopropylfluorophosphate (DFP), the thrombogenicity appeared to decrease slightly in all three tests, but was not completely eliminated. Heparinagarose was found to adsorb the material(s) responsible for shortening the NAPTT and TGt50, and it also adsorbed factor IX. Gel filtration studies clearly separated the three activities into different sizes; the TGt50 activity was the largest with an apparent molecular weight of 1 × 106, FEIBA activity was intermediate with an apparent molecular weight of 1.30 × 105 and NAPTT activity was the smallest with an apparent molecular weight of 7 × 104. Our results show that the thrombogenicity measured by these three tests may be related to the presence of physically distinct materials.
Haemolytic Anaemia Associated with Glucosephosphate Isomerase (GPI) Deficiency in a Black South African Child
Authors: Cayanis Eftihia , Penfold Grace K. , Freiman Ida , Mac Dougall Lorna G.
Published: March 2008

Summary. Haemolytic anaemia in a Black South African child was found to be associated with reduced glucosephosphate isomerase activity in the red cells. Apart from the haemolytic anaemia, there was no other clinical evidence of dysfunction. Family studies pointed to an autosomal recessive mode of inheritance, with the symptomatic homozygous propositus having a marked enzyme deficiency and the asymptomatic heterozygotes showing intermediate levels of activity. Biochemical characterization showed that, apart from being thermolabile, the electrophoretic mobility and the kinetic properties of the variant enzyme were similar to those of the normal wild type.
Differentiation Linked Expression of p28,33 (Ia‐like) Structures on Human Leukaemic Cells
Authors: Janossy George , Goldstone Antony H. , Capellaro David , Greaves Melvyn F. , Kulenkampff Jörg , Pippard Martin , Welsh Ken
Published: March 2008

Summary. A differentiation linked expression of ‘Ia‐like’ (B cell associated) structures was detected by heterologous antisera against p28,33 antigen on human leukaemic cells in immunofluorescence test. A double fluorochrome immunofluorescence test was used to distinguish Ia+ (p28,33, +) leukaemic cells from B lymphocytes. Leukaemic cells with varying intensities of anti‐p28,33 staining were separated using the Fluorescence Activated Cell Sorter. The results indicate that p28,33 is very strongly expressed on undifferentiated blast cells from Philadelphia chromosome (Ph1) positive ‘lymphoid’ blast crisis and non‐T,non‐B acute lymphoid leukaemia. In Ph1 positive myeloid blast crisis and in acute myeloid leukaemia the p28,33 antigen was only moderately strongly expressed on myeloblasts and virtually absent on promyelocytes. The p28,33 antigen was also absent on the relatively more mature cells during the chronic phase of chronic myeloid leukaemia. Non‐leukaemic samples such as bone marrow samples from fetus, and from patients with severe megaloblastic anaemia or myeloid hyperplasia (due to hypersplenism in Felty syndrome) were also found to contain p28,33 positive (and membrane immunoglobulin negative) immature blast cells.

On the basis of these observations the hypothesis is proposed that ‘Ia‐like’ antigens on leukaemic cells reflect the origin of these leukaemias from haemopoietic stem cells which are themselves Ia positive. The corollary of this view is that ‘Ia‐like’ membrane structures may play an important role not only in immunological responses of B cell, but also in the regulation of early haemopoietic cell differentiation.
The Significance of the Philadelphia Chromosome in Acute Lymphoblastic Leukaemia: a Report of Two Cases
Authors: Gibbs T. J. , Wheeler M. V. , Bellingham A. J. , Walker S.
Published: March 2008

Summary. Two cases of Philadelphia chromosome (Ph1) positive acute lymphoblastic leukaemia are reported, both of which lost the Philadelphia chromosome during remission. In one patient remission of the acute lymphoblastic leukaemia continued but classical Ph1 positive chronic granulocytic leukaemia developed. In the other patient relapse of the acute lymphoblastic leukaemia occurred associated with the return of the Ph1 chromosome. The evidence suggests that the chromosome aberration occurred in a pluripotential stem cell, which in one case proliferated along both a lymphoid cell line and a myeloid cell line. Both cases responded well to conventional therapy for acute lymphoblastic leukaemia.
Hairy Cell Leukaemia: Seven Cases with Probable B‐Lymphocytic Origin
Authors: Zidar B. L. , Winkelstein A. , Whiteside T. L. , Shadduck R. K. , Zeigler Z. , Smith W. I. , Rabin B. S. , Krause J. R. , Lee R. E.
Published: March 2008

Summary. Mononuclear cells from seven patients with hairy cell leukaemia were examined for features suggestive of either a lymphocytic or monocytic origin. Immunofluorescent staining of both methanol fixed and incubated cells, using monospecific antisera, revealed a predominant cell‐associated immunoglobulin in each case. Three were positive for mu and kappa chains, two for gamma and kappa chains, one for delta and kappa chain determinants and one reacted only with antigamma chain serum. Formation of EAC rosettes, a feature of both B lymphocytes and monocytes, was variable. T cells, as judged by E rosettes, were not elevated in any patient. Phytohaemagglutinin reactivity was normal in six and depressed in one case. With the exception of minimal activity in assays for glass adherence and latex particle phagocytosis, none of the cells showed features typical of monocytes. Hairy cells were negative by peroxidase stain and lacked the electron microscopic characteristics of monocytes. They did not react in either rosette or phagocytic assays with anti‐A or anti‐D coated erythrocytes nor did they elaborate granulocyte colony stimulating factor, a monocyte‐derived in vitro granulopoietin. Although unequivocal classification of these abnormal cells is not possible, the data strongly suggests that this represents a variant of a B lymphocytic neoplasm.
Nuclear Labelling of Leukaemic Blast Cells with Tritiated Thymidine Triphosphate in 35 Patients with Acute Leukaemia
Authors: Wantzin Gunhild Lange
Published: March 2008

Summary. A new technique which detects the presence of DNA polymerase and primer‐template DNA by measuring the in vitro incorporation of [3H]thymidine‐5‐triphosphate (3H‐TTP) into nuclei of leukaemic blast cells (LBC) was used in 35 patients with acute leukaemia. The 3H‐TTP labelling index (3H‐TTP LI) exceeded the fraction in DNA synthesis by a factor 1.4–24.3. The values of 3H‐TTP labelling in the bone marrow always exceeded those obtained in the blood. In addition 10 normal bone marrows were studied; here, the 3H‐TTP LI either exceeded or equalled the fraction of the proliferative pool in DNA synthesis.
Ferritin in Human Liver Cells of Homozygous Beta‐Thalassaemia: Ultrastructural Observations
Authors: Iancu T. C. , Neustein H. B.
Published: March 2008

Summary. Homozygous beta‐thalassaemia is a disease in which there is a progressive iron overload from infancy to death in early adulthood. Liver biopsies from 10 patients in various stages of this disease were examined by electron microscopy. A number of round or oval lysosomal structures, containing lamellae different from myelin figures, were seen in all patients, including those with minimal iron overload. Ferritin molecules were seen either in relationship with the lamellae forming arrays, or in paracrystalline arrangement, or with no organized form. There were practically no ferritin molecules in sub‐cellular compartments other than cell sap and lysosomes. The density of cell sap ferritin was constant beyond infancy, but the number of iron‐laden lysosomes increased with age.

The stages in the process of iron seclusion, seen even in advanced phases of iron overload, are described. Ferritin is thought to accumulate in lysosomes by a transmembraneous movement, but other explanations are considered.
Identification of Sources of Inter‐Laboratory Variation in Factor VIII Assay
Authors: Kirkwood T. B. L. , Rizza C. R. , Snape T. J. , Rhymes I. L. , Austen D. E. G.
Published: March 2008

Summary. A repeated finding of national and international collaborative studies of standard factor VIII preparations has been that systematic differences exist between laboratories in their measurement of the relative activities of the same pairs of factor VIII preparations.

A workshop meeting was held at the Oxford Haemophilia Centre during 23–26 November 1976 to investigate which of the possible sources of variation between laboratories were responsible. Participants from 16 British laboratories (nine using one‐stage assays and seven using two‐stage assays) performed a total of 273 valid assays using three freeze‐dried preparations of differing purity (a plasma, an intermediate and a high purity concentrate). The results of assay with each participant using their normal system established that, if the participants were a representative cross‐section, approximately one‐third of one‐stage laboratories would show a systematic difference from the overall mean of at least 16% with a corresponding figure for the two‐stage laboratories of 9%. Various features of the assay systems were then modified in a controlled series of experiments. The results showed conclusively that (i) differences between reagents accounted for most of the variation between laboratories, and (ii) the two‐stage assays were, on average, detecting relatively more activity in the more purified preparations than the one‐stage assays. The results also suggested that the use of buffer or citrate‐saline as opposed to haemophilic plasma for the initial dilution of concentrates did not affect the assay results.
HLA Antigens in Addisonian Pernicious Anaemia: Absence of an HLA and Disease Association
Authors: Wright J. P. , Callender S. T. E. , Grumet F. C. , Payne R. O. , Taylor K. B.
Published: March 2008

Summary. An examination of HLA antigens in 72 unrelated Caucasian subjects with pernicious anaemia (PA) has revealed no significant association of any HLA‐A or B genes with the disease. These data do not confirm the previous reports in the literature which had suggested an increased frequency of the B7 and/or A3 antigen among patients. In addition, the study of four families, each with two or more PA patients, does not support close linkage between disease susceptibility or autoantibody formation and the HLA locus. These data suggest that genes in or near the HLA region may not significantly affect susceptibility to PA.
The Role of Endocytosis in Transferrin Uptake by Reticulocytes and Bone Marrow Cells
Authors: Hemmaplardh Deeporn , Morgan E. H.
Published: March 2008

Summary. The aims of the work described in this paper were to investigate the process by which transferrin enters immature erythroid cells, to determine whether entry of transferrin into the cells is required for iron utilization by the cells and to see whether substances in the medium surrounding the cells enter the cells along with transferrin. The kxperiments were performed using rabbit transferrin, rabbit reticulocytes and rabbit bone marrow cell suspensions. The process of transferrin uptake was studied by electron microscope techniques using autoradiography of 125I‐labelled transferrin or transferrin conjugated to ferritin or horseradish per‐oxidase. The results confirmed that transferrin enters reticulocytes and showed that the protein also enters nucleated erythroid cells of the bone marrow but very little is taken up by platelets, cells of the white cell series in the bone marrow or peripheral blood or mature erythrocytes. The two stages of transferrin uptake by erythroid cells, initial absorption and progressive uptake, were shown to be represented morphologically by binding of transferrin to the outer cell membrane, probably to specific receptors, and entry into the cells by endocytosis, respectively. Reticulocytes were unable to utilize 59Fe for haem synthesis when the 59Fe was on transferrin which was bound to agarose, Enzacryl AA or Latex beads so that it would not enter the cells. The uptake of transferrin by reticulocytes was not accompanied by uptake of 14C‐polyethylene glycol dissolved in the medium.
Thyroid Hormones Stimulate Erythropoiesis in Vitro
Authors: Golde D. W. , Bersch N. , Chopra I.J. , Cline M.J.
Published: March 2008

Thyroid hormones have important effects on erythropoiesis in man and animals. We performed in vitro culture studies with murine and human bone marrow in order to define the interaction of these hormones with erythroid and granulocyte‐monocyte progenitor cells. The methylcellulose clonogenic assay was used with the appropriate addition of erythropoietin or colony‐stimulating activity. L‐Thyroxine, D‐thyroxine and L‐triiodothyronine potentiated erythropoietin‐stimulated erythroid colony formation in concentrations of 50–100 ng/ml. These hormones had no effect on granulocyte‐monocyte colony formation at concentrations up to 500 ng/ml. Testing of various thyroid analogues showed no clear correlation between potentiation of erythropoiesis and known calorigenic potency. Reverse triiodothyronine also had potentiating activity in this system. The active thyroid hormones stimulated erythroid colony formation at several concentrations of erythropoietin but could not substitute for erythropoietin. These data suggest that thyroid hormones have a direct effect on erythroid precursor proliferative capacity, a finding which may have relevance to the mechanism of erythropoietic dysfunction in human thyroid disease.
The Lymphocyte as a Marker of Past Nutritional Status: Persistence of Abnormal Lymphocyte Deoxyuridine (dU) Suppression Test and Chromosomes in Patients with Past Deficiency of Folate and Vitamin B12*
Authors: Das Kshitish C. , Herbert Victor
Published: March 2008

In short‐term suspension cultures of bone marrow cells or PHA‐stimulated lymphocytes from normal subjects, non‐radioactive deoxyuridine (dU) suppresses the incorporation of radioactive thymidine ([3H]TdR) or its analogue, [125I]deoxyuridine ([125I]Udr), into DNA. This normal suppression by deoxyuridine (dU) is impaired in both of these cell systems from patients with deficiency of folate or vitamin B12, and corrected by the appropriate vitamin. Patients with megaloblastic anaemia due to deficiency of vitamin B12 or folate were studied before and after treatment. When treatment had returned to normal the bone marrow morphology and the serum and red cell vitamin levels, then the dU suppression test and chromosomal changes in the bone marrow were also corrected. However, the dU suppression test and chromosomal changes remained abnormal in lymphocytes as long as 84 d after therapy. These abnormal lymphocyte dU suppression tests were corrected by the appropriate in vitro additions of folic acid, methylfolate and vitamin B12, depending on the vitamin deficiency present before therapy. These studies suggest that an abnormal lymphocyte dU suppression test corrected by the appropriate vitamin in vitro, and characteristic chromosome abnormalities in lymphocytes, when these are absent in the bone marrow, indicate past deficiency of vitamin B12 or folate. These changes can be used for retrospective diagnosis of these deficiencies in patients treated by ‘shotgun’ therapy. They further support the concepts that circulating unstimulated lymphocytes: (1) do not incorporate appreciable amounts of vitamin B12 or folic acid; (2) reflect the vitamin status of the patient at the time the lymphocytes were generated; and (3) cannot replace bone marrow in dU suppression tests aimed at diagnosis of current marrow and other non‐lymphocyte cell line nutrient status. These studies add to the evidence that selective nutrient deficiency may occur in one but not another cell line in the same person, and point to the need for more studies on factors affecting nutrient delivery, uptake, and utilization by various human cell lines. These studies also provide a new approach to evaluation of circulating lymphocyte age.
Subunits A and S Inheritance in Four Families with Congenital Factor XIII Deficiency
Authors: Barbui T. , Rodeghiero F. , Dini E. , Mariani G. , Papa M. L. , De Biasi R. , Murillo R. Cordero , Umana C. Montero
Published: March 2008

Previous studies of the inheritance of the two molecular subunits of fibrin stabilizing factor (factor XIII) refer to isolated cases. The present work investigates the hereditary mode of transmission of subunits A and S, measured by the Laurell technique, in seven homozygotes and in 29 heterozygotes belonging to four families with factor XIII deficiency. The results indicate that the homozygotes were devoid of immunologically identifiable A subunit, whereas the heterozygotes could be identified by measuring this protein. The subunit S has been found to be decreased both in homozygous and in heterozygous patients, so that it seems that the two subunits, even if their synthesis is controlled by different genes, are genetically related. The mode of transmission of this disorder, supported by quantitative determinations of plasma subunit A, is autosomal recessive.
Folate‐dependent Serine Synthesis in Lymphocytes from Controls and Patients with Megaloblastic Anaemia: the Effect of Therapy
Authors: Tikerpae J. , Chanarin I.
Published: March 2008

The utilization of [14C]formate for serine synthesis by lymphocytes was impaired in all the patients with pernicious anaemia and in 70% of other patients with megaloblastic anaemia. In pernicious anaemia this was corrected by vitamin B12 therapy in 48 h but not by folate therapy although patients given folate showed a satisfactory haematological response.
Neutrophil and Eosinophil Granulocytes in Bacterial Infection: Sequential Studies of Cellular and Serum Levels of Granule Proteins
Authors: Venge P. , Strömberg A. , Braconier J. H. , Roxin L.‐E. , Olsson I.
Published: March 2008

The intraneutrophilic concentrations of lactoferrin, myeloperoxidase, collagenase and chymotrypsin‐like cationic proteins were measured sequentially during acute bacterial infection. The serum levels of lactoferrin and myeloperoxidase were also followed as well as the ‘eosinophil’ cationic protein as a marker for eosinophil leucocytes. During the early course of infection there was a profound but reversible decrease of intraneutrophilic lactoferrin. The levels of cellular collagenase and chymotrypsin‐like cationic proteins also tended to decrease reversibly during day 2–8 in most cases; myeloperoxidase levels were normal except for two cases. Serum myeloperoxidase and lactoferrin correlated with blood neutrophil counts. In spite of the absence of peripheral eosinophils the ‘eosinophil’ cationic proteins of serum were increased on the first day of infection, which may reflect increased eosinophil turnover.
The Relationship between Serum Viscosity, Hypervolaemia and Clinical Manifestations Associated with Circulating Paraprotein
Authors: Russell J. A. , Powles R. L.
Published: March 2008

In 44 of 56 patients with malignant paraproteinaemia the relationship of serum viscosity, plasma volume (PV), estimated blood volume (BV) and clinical findings was investigated and in the remaining 12 patients one or more of these parameters was studied. There was a correlation between increased PV and serum viscosity (r= 0.66, P<0.001). The retinopathy characteristic of the hyperviscosity syndrome (HVS) was always associated with serum viscosity ≥3.8 and with hypervolaemia. The evidence that the retinal changes were related more closely to serum hyperviscosity than to hypervolaemia is twofold. First, there was a considerable overlap in the degree of BV expansion, but not serum hyperviscosity, in patients with and without retinopathy. Second, the retinopathy in a patient with Waldenström's macroglobulinaemia (WM) improved after plasma exchange with little change in BV. While bleeding was often associated with retinopathy seven patients with multiple myeloma (MM) had no retinopathy, viscosities ≥4.3 and BV in the same range as patients with no clinical signs. Haemorrhage in such patients may thus be unrelated to the property of the paraprotein responsible for hyperviscosity. Measurements of PV in a patient with WM having repeated plasma exchanges for bleeding suggested that hypervolaemia may contribute to haemorrhage in some patients. It is suggested that the term ‘HVS’ should be restricted to patients with retinopathy and that hypervolaemia should be considered a characteristic feature of this syndrome.
The Ultrastructural Localization of Factor VIII‐Antigen in Human Platelets, Megakaryocytes and Endothelial Cells Utilizing a Ferritin‐labelled Antibody
Authors: Piovella F. , Nalli G. , Malamani G. D. , Majolino I. , Frassoni F. , Sitar G. M. , Ruggeri A. , Dell'Orbo C. , Ascari E.
Published: March 2008

By means of electron microscopy combined with the use of monospecific anti‐factor VIII‐antigen (VIIIR:AG) antibodies conjugated to ferritin, the subcellular localization of VIIIR:AG in platelets, megakaryocytes and in endothelial cells has been established. The reported results suggest the possibility that the megakaryocyte is able to synthesize VIIIR:AG and also to secrete it by means of a microcanalicular system similar to that present in the endothelial cell. Platelets may derive their VIIIR:AG content partly from the megakaryocyte and partly from the plasma.
Factor VIII Coagulant Activity in an African Population in Relation to a Recognized Standard*
Authors: Essien E. M. , Ayeni O.
Published: March 2008

The normal range of factor VIII coagulant activity (derived from log potency ratio) in some sections of the Nigerian population has been established at 0.65–5.55 iu/ml with a geometric mean of 1.90 iu/ml. This was determined against an acceptable standard (MRC Human 68/413 with activity 0.66 iu/ml). The distribution of the potency ratio was log normal. The level was not affected by age or an abnormal haemoglobin (Hb A + S or A + C). The mean activity in females was significantly higher than the mean value in male subjects. With the use of a stable standard, our results show that the conclusions of some previous studies in respect of some of the parameters such as population distribution but which did not use a recognized standard, were valid. Within the age limits of our subjects, age did not affect the population level of factor VIII coagulant activity.
SOME ASPECTS OF RECENT ADVANCES IN THE USE OF BLOOD CELL COMPONENTS
Authors: Schiffer C. A.
Published: March 2008

Intensive research and major technical advances in the last decade have resulted in a remarkable improvement in transfusion supportive care which has significantly contributed to the increased survival now achievable in patients with leukaemia and aplastic anaemia. Platelet transfusions are now available in almost all hospitals, while granulocyte transfusions, previously used only in cancer centres, are now being dispensed by many regional blood transfusion centres in the United States. A number of reviews dealing with donor selection and safety, collection methodology and clinical guidelines for cell component use have been published in the last few years
The Blast Crisis of Chronic Granulocytic Leukaemia: Megakaryoblastic Nature of Cells as Revealed by the Presence of Platelet‐Peroxidase—A Cytochemical Ultrastructural Study
Authors: Breton‐Gorius J. , Reyes F. , Vernant J. P. , Tulliez M. , Dreyfus B.
Published: March 2008

The origin of cells in the blast crisis of some cases of chronic granulocytic leukaemia (CGL) remains controversial. Difficulties arise from the lack of cytochemical characteristics of differentiation. This report concerns the nature of cells in the blast crisis of a case of CGL in which blast cells exhibited an undifferentiated or lymphoid appearance by light and electron microscopy. The majority (90%) of such cells contained a peroxidase in the endoplasmic reticulum distinct from myeloperoxidase. In addition, some micromegakaryocytes could be recognized among the peroxidase reactive cells, by the presence of typical granules and demarcation membranes. Since this peroxidase exhibited identical characteristics to that of normal megakaryocytic precursors, these blast cells could be identified as megakaryoblasts.

These data emphasize the possible megakaryoblastic nature of cells occurring in other cases of CGL blast crisis.
Chromosomes in Ph1‐Positive Chronic Granulocytic Leukaemia
Authors:   FIRST INTERNATIONAL WORKSHOP ON CHROMOSOMES IN LEUKAEMIA
Published: March 2008

Information about the type, frequency, and clinical significance of chromosome abnormalities in leukaemia has been reported from a number of laboratories located in various countries. A small international Workshop was organized to examine the combined data of the participating investigators for chronic granulocytic leukaemia (CGL) and acute nonlymphocytic leukaemia (ANLL). In this report the results of the analysis of chromosomes in CGL are presented; the results of a similar analysis in ANLL are presented in the subsequent paper.

The Workshop addressed itself primarily to: (I) the incidence of various karyotypic abnormalities, in the different phases of Ph1‐positive CGL, based on the data of the various members of the group; (2) a comparison of the findings with those in the literature; and (3) the possible utilization of unique cytogenetic phenomena as predictive parameters for the blastic phase and other clinical aspects of CGL.
Myeloid Karyotype and the Malignant Phase of Chronic Granulocytic Leukaemia
Authors: Lilleyman J. S. , Potter A. M. , Watmore A. E. , Cooke P. , Sokol R. J. , Wood J. K. ,   THE TRENT REGION AND ASSOCIATED HOSPITALS WORKING PARTY ON ADULT LEUKAEMIA
Published: March 2008

During the course of a 30 month study period, 26 patients with typical chronic granulocytic leukaemia (CGL) developed karyotype abnormalities in addition to the Philadelphia (Ph1) chromosome. All cases had received at least 14 months continuous low dose busulphan, and the chromosome changes were found before clinical transformation in six patients and at the time this occurred in 20.

Survival following the discovery of these additional abnormalities was short, with a median of 11 weeks for the whole group. Trisomy 8 was the commonest additional chromosome abnormality, but no one karyotype change was clearly associated with shorter survival than another.

23 of the 26 have died as a direct result of their disease, forming part of a total of 40 deaths from typical CGL encountered during the study period where karyotype analyses were performed during the terminal stages of the disease. Of these 40, only two (5%) patients showed no chromosome abnormalities extra to the Ph1 prior to death. (The 17 non‐study patients who died were similar in all respects to the 23 from the study group, but had no prior chromosome studies performed during the chronic phase of the disease.)

It is suggested that an expanding aneuploid or pseudodiploid clone arising from the leukaemic cells during the benign phase of CGL can be used to mark the sometimes ill‐defined onset of the malignant phase in all but a small proportion of cases.
Circulating Inhibitors of Granulopoiesis in Patients with Aplastic Anaemia
Authors: Gordon M. Y.
Published: March 2008

Summary. An inhibitor of granulopoiesis has been detected in the serum of aplastic patients by its ability to reduce colony formation by normal human marrow cells in culture. The inhibitory activity in the serum was abolished when a patient was treated with azathioprine and exposure of aplastic marrow to antithymocyte globulin (ATG) in vitro released the colony forming cells from suppression. The clinical findings in three of the patients studied support the idea that the inhibitory activity detected in vitro is associated with normal marrow suppression in vivo.
Platelet‐Associated Coagulation Factors: Immunological Detection and the Effect of Calcium
Authors: Giddings J. C. , Shearn S. A. M. , Bloom A. L.
Published: March 2008

Normal platelets were examined by conventional coagulation methods and specific immunological techniques for the presence of platelet‐associated coagulation factors. Platelets washed in buffer containing calcium chloride gave different results from those washed in the absence of calcium. Factor XI was not detected in washed platelets by any technique. Factor II and factor X were removed from platelets by washing in calcium‐free buffer but not by calcium‐containing buffer. Procoagulant factor VIII was readily removed or inactivated by washing in the presence or absence of calcium, but factor VIII related antigen was consistently demonstrated after multiple washes in either buffer. Factor V activity and antigen was detected in platelet suspensions after multiple washes although the presence of calcium in the washing buffer gave higher factor V assays and stronger immunological reactions than when calcium‐free buffer was used. The results are consistent with the presence of calcium dependent binding sites in platelets which may have a role in the enhancement of factor V and phospholipid reactions.
A Possible Contributory Role of the Platelet in the Formation of Plasma Factor XIII
Authors: Rider Diane M. , McDonagh R. P. , McDonagh Jan
Published: March 2008

Factor XIII is a proenzyme found in plasma and platelets which, when activated, catalyses the formation of ε‐(γ‐glutamyl)lysyl bonds in fibrin. Plasma factor XIII is composed of two different polypeptides, a and b, whereas platelet factor XIII has only a subunit. Factor XIIIa, the active enzyme, from both plasma and platelets is enzymatically the same. Platelet factor XIII is synthesized in megakaryocytes and accounts for 50% of the total factor XIII activity in blood. In this study the role of platelet factor XIII in the formation of plasma factor XIII was investigated. The chemotherapeutic agents—quinidine, vincristine, methotrexate and cyclophosphamide—were used to induce thrombocytopenia in rats. Platelet count, plasma factor XIII activity, and fibrinogen concentration were determined along with other haematological parameters. With all four drugs there were concomitant decreases in plasma factor XIII activity along with decreases in the platelet count. Throughout the experiment, fibrinogen concentrations remained within pretreatment levels. In the saline‐treated control animals, the platelet count and plasma factor XIII activity did not significantly differ from the pre‐treatment levels. These observations suggest that the low levels of factor XIII achieved in the treated groups resulted from the effects of the chemotherapeutic agents on thrombocytopoiesis. Statistical analyses of the data indicated a high correlation between platelet count and plasma factor XIII activity. From this study, it is apparent that platelet factor XIII could be a source of a subunit for plasma factor XIII.
Use of an Antiserum Specific for Human T Lymphocytes as a Diagnostic Reagent: Comparison with E Rosette Formation and Mitogenic Response to PHA
Authors: Zola H. , Valdimarsson H.
Published: March 2008

An antiserum with specificity for human T lymphocytes was evaluated as a diagnostic reagent in the clinical immunology laboratory. The antiserum was used in indirect immunofluorescence to detect T cells in blood samples from normal controls and patients with various disorders involving the immune system. The same samples were also examined using two established tests for T cells, the rosette reaction with sheep red blood cells (E rosette assay) and the proliferative response to phytohaemagglutinin (PHA). The antiserum and the rosette assay detected approximately the same numbers of T cells in controls, but the antiserum indicated T cell deficiency in several patients who appeared normal by resetting or PHA reactivity. PHA stimulation in autologous plasma was depressed in a further group of patients who had normal T cell numbers by the other two tests. In vitro experiments with normal lymphocytes indicated that the E‐rosette receptor was distinct from determinants detected by the T‐cell specific antiserum. The immunofluorescence test with the anti‐T serum provides an additional assay which appears to be more sensitive than the other two tests in detecting certain cases of T lymphocyte deficiency.
Metabolic Disorders in Acute Myeloid Leukaemia
Authors: Mir M. Afzal , Delamore I. W.
Published: March 2008

SUMMARY Metabolic balance studies were carried out in 17 utiselected patients with acute myeloid leukaemia. Widespread metabolic disturbances were observed. Serum Na fell below 135 mmol/l in 14 patients (82%) and 11 patients (64%) developed hypokalaemia. An increased osmolal clearance caused by a release of electrolyte and blast cell waste (i.e. urea, urate, etc.) during chemotherapy appeared to be the principle cause of natriuresis and hyperkaluria. Seven patients had pro‐teinuria before and eight others developed it during antileukaemic therapy. Nine patients (53%) developed proximal renal tubular dysfunction with aniinoaciduria, hyperphosphaturia and incomplete reabsorption of urate. No significant relation was found between this widespread glotnerulo‐tubular dysfunction and lysozymuria. We suggest that antileukaemic drugs release unidentified substances from blast cells which are toxic to the kidney. Metabolic alkalosis in six patients (35%) was probably related to volume depletion and hypokalaemia, while two patients developed acidaemia with the onset of renal failure. Hypocalcaemia in seven patients (41%) had a multifactorial basis: hyperphosphaturia, septicaemia, malnutrition and cytotoxic drugs were among the probable causes.
A Comparative Study of the Buoyant Density Distribution of Normal and Malignant Lymphocytes
Authors: Huber Christoph , Zier K. , Michlmayr G. , Root H. , Nilsson K. , Theml H. , Lutz D. , Braunsteiner H.
Published: March 2008

SUMMARY Density distribution patterns of normal and malignant lymphocytes were compared following centrifugation to equilibrium on linear density gradients.

For normal lymphocytes differences in the distribution patterns were observed between: (1) B and T cells, (2) central and peripheral cells, and (3) resting and activated cells. The findings suggested that cell density is determined by cell lineage, the degree of differentiation and the stage of functional activation.

Marked differences in the density distribution profiles were also observed among certain types of morphologically distinguishable lymphoproliferations. To some extent density analyses enabled the discrimination between CLL, follicular lymphomas and lymphoblastic lymphomas as well as between O‐ALL and T‐ALL. Density profiles of malignant lymphocytes failed to disclose any features specific for malignancies. But they revealed some similarities with distinct subsets of normal lymphocytes, i.e. between: (1) CLL and bone marrow lymphoid cells, (2) follicular lymphomas and follicular centre cells, and (3) lymphoblastic lymphomas and activated lymphocytes. These findings arc further evidence supporting the hypothesis that the malignant trarisformation of phenotypicalIy different lyrnplioproliferations takes place at different levels of lymphocyte differentiation.
Ways of Expressing Results of Human Bone Marrow Progenitor Cell Culture
Authors: Parmentier C. , Droz J. P. , Tubiana M.
Published: March 2008

SUMMARY In human bone marrow culture studies the number of granulocyte progenitor cells (CFUc) is expressed as the numbcr of colonies per 105 incubated, nucleated cells. However, this provides little information on cell proliferation kinetics of granulopoiesis. The ratio of progenitors (CFUc) to end cells (metamyelocytcs or granuloc)‐tcs) is related to the effectiveness of granulopoiesis and this allows an estimate to be made of an amplification factor within the maturing and proliferating compartments. We measured an index equal to the number of CFUc per 105 nietamyelocytes incubated in both normal individuals and in patients. The index falls when a large number of granulocytes is produced by each progenitor cell. This was observed in many patients with a neutropenia. An estimation of the total number of mctamyelocytes is based on the ratio of the number of metamyelocytes to the number of blood granulocytes. This allows an estimate of the total number of CFUcs, which is of clinical interest.
Collection, Cryopreservation and Subsequent Viability of Haemopoietic Stem Cells Intended for Treatment of Chronic Granulocytic Leukaemia in Blast‐Cell Transformation
Authors: Goldman J. M. , Th'ng K. H. , Park D. S. , Spiers A. S. D. , Lowenthal R. M. , Ruutu T.
Published: March 2008

Summary. . We have stored at – 196°C peripheral blood buffy coat (BC) and bone marrow (BM) cells collected from 47 patients with chronic granulocytic leukaemia in the chronic phase. Dimethyl sulphoxide (DMSO) 10% was used as cryoprotective agent. As these cells include CFUc and probably pluripotential stem cells they may be transfused as part of the management of patients who enter blast cell transformation. The mean numbers of nucleated cells collected and stored per procedure was about 9 times greater for BC collections than for BM harvests (106 ± 49 (SD) × 109 versus 11.9 ± 6.6 × 109 respectively). Agar CFUc assay showed that stored cells may remain viable for up to 5 years. Since in vitro studies showed that CFUc proliferation is not inhibited by low concentrations of DMSo the removal of all DMSO during cell reconstitution before transfusion may not be necessary. If autologous BC cells are capable of repopulating the BM of patients treated for CGL in blast cell transformation the routine collection and storage of BC rather than BM cells may be desirable for all newly diagnosed patients.
Plasma Clearance and Renal Excretion of Erythrocyte Cryoprotectant Hydroxyethylated Amylopectin
Authors: Milton Mishler John , Parry Ernest S. , Petrie Aviva
Published: March 2008

Summary. The value of low molecular weight‐hyroxyethylated amylopectin (cryo‐HES) as an extracellular cryoprotectant has been demonstrated in vitro. It is important that details of the intravascular persistence and urinary excretion be determined to compare with data already available as other grades of HES and with data on transfusion of cryo‐HES cryoprotected blood. Following a single 400 ml (14% solution) infusion in man, the intravascular clearance of cryo‐HES was well described mathematically by the equation: y=3.30+6.49e‐0.15x. The plasma concentration of cryo‐HES fell to half its peak value in approxiamately 9.6 h. Approximately 20% of the total infused cryo‐HES was excreted in the urine during the first post‐injection hour, and 50% by 72 h. The ESR was not altered significantly by the presence of this material. The present study indicates that cryo‐HES is eliminated rapidly and may thus be safe for transfusion to recipients of frozen blood.
Non‐Specific Serum Iron in Thalassaemia: an Abnormal Serum Iron Fraction of Potential Toxicity
Authors: Hershko Chaim , Graham Gary , Bates George W. , Rachmilewitz Eliezer A.
Published: March 2008

Summary.Iron binding in the sera of 35 patients with β thalassaemia major and intermedia was studied. In patients receiving regular blood transfusions since infancy transferrin was completely saturated and about 2.7‐7.I μmol/1 of the serum iron could be removed by dialysis or ultrafiltration in the presence of a chelating agent or by filtration on DEAE‐Sephadex‐catecholdisulphonic acid columns. In contrast, less than 1.0 μmol/1 of transferrin bound iron was removed when subjected to the same procedures. The non‐specific iron of thalassaemic sera could no longer be demonstrated after incubation with normal serum. These findings indicate that non‐specific iron is a chelatable compound which is readily available for transferrin binding. In view of the known toxicity of unbound iron, its identification in thalassaemic sera might be of relevance to the pathogenesis of tissue damage and the protective effect of iron chelating therapy in this disease.
Different Requirements for Intrinsic Factor‐Xa Forming Activity and Platelet Factor 3 Activity and their Relationship to Platelet Aggregation and Secretion
Authors: Walsh Peter N.
Published: March 2008

Summary. Platelets provide coagulant activity in part by promoting two essential reactions of intrinsic coagulation: (I) factor‐X activation by a complex of factor IXa, factor VIII, calcium and platelets (intrinsic factor‐Xa forming activity or XaFA); and (2) prothrombin activation by a complex of factor Xa, factor V, calcium and platelets (platelet factor 3 activity or PF3A). We found that a chloroform extract of acetone‐dried brain had about half the XaFA of a petroleum ether extract of brain and over twice the PF3A of the petroleum ether extract; and that when purified phospholipids other than phosphatidylserine (PS) were added to PS, the resultant activity, compared with PS alone, was reduced in the assay for XaFA and enhanced in the assay for PF3A. In experiments with washed platelets stirred with collagen XaFA developed rapidly (100% activity at 30 s), well before maximum [14C]5HT release (2‐4 min) and decayed to 15% at 10 min. In contrast, PF3A developed slowly (100% activity after 20 min) well after maximal aggregation and release. Maximal XaFA developed in unstirred platelet suspensions incubated with collagen, whereas PF3A did not become available unless platelet suspensions were stirred and underwent secretion. Platelets stirred with ADP released [14C]5HT and developed PF3A but not XaFA. Both [14C]5HT release and PF3A by collagen were inhibited by p‐chloromercuriphenylsulphonate, indomethacin and a combination of prostaglandin E1 and RA 233, but XaFA was only minimally affected by these inhibitors. In contrast concanavalin A inhibited collagen‐induced PF3A and XaFA in dose‐dependent fashion but had no effect on [14C]5HT release. Both platelet coagulant activities and [14C]5HT release were inhibited by aspirin, EDTA and a combination of antimycin A and 2‐deoxy‐D‐glucose. These results indicate that the biochemical determinants of XaFA and PF3A are different. PF3A developed only when aggregation and release occurred whereas XaFA was independent of aggregation and release.
Discrepancy between One‐stage and Two‐stage Assay of Factor VIII:C
Authors: Kirkwood T. B. L. , Barrowcliffe T. W.
Published: March 2008

Summary. Two methods (one‐stage and two‐stage) are commonly used for the assay of factor VIII clotting activity (VIII:C). We present collected data from seven separate studies of VIII:C assay which show that these methods do not give the same result when comparing concentrates and plasma. On average, two‐stage assays detect 20% more VIII:C activity in concentrates as compared to plasmas than do one‐stage assays.
Splenic Red Cell Pooling: A Diagnostic Feature in Polycythaemia
Authors: Bateman S. , Lewis S. M. , Nicholas A. , Zaafran A.
Published: March 2008

Summary. Total red cell volumes and splenic red cell pools were measured in 31 patients with polycythaemia. 22 had polycythaemia vera (PV), 12 of whom had clinically detectable splenomegaly, and nine patients had secondary polycythaemia (PS). The mean red cell pool was 192.8ml (SD 126.6) in PV (all cases), and 130.9 ml (SD 28.4ml) in PV without splenomegaly; it was 61.1 ml (SD 8.3 ml) in PS. When expressed relative to spleen size (in cm), differences were even more striking: PV (all cases)—mean 13.7 ml/cm (SD 4.3); PV without splenomegaly—mean 12.7 ml/cm (SD 2.2); PS—mean 6.6ml/cm (SD 1.2). Measurement of splenic red cell pool thus appears to be valuable diagnostic tool for distinguishing between PV and PS. The findings point to the presence in PV of a splenic structural abnormality which is not simply an effect of the inceased circulating red cell mass.
Fetal Haemoglobin Production and the Sickle Gene in the Oases of Eastern Saudi Arabia
Authors: Pembrey M. E. , Wood W. G. , Weatherall D. J. , Perrine R. P.
Published: March 2008

Summary. Fetal haeamoglobnin(hb F) levels have been measured in 137 normal (AA) subjects, 109 with the sickle‐cell trait (AS) and 237 with sickle‐cell anaemia (SS) from the oasis population of Eastern Saudi Arabia. In addition the proportion of F‐cells has been estimated in 71 AA, 51 AS and 34 SS subjects. The mean Hb F% (and the range of F‐cells %) were: AA 0.77 (0.3–18), AS 1.38 (2.3–43) and SS 25.56 (33–98). The distribution of Hb F was always heterocellular. The sickle‐cell trait showing rasised Hb F and F‐cells. Whilst the normal Arabs and those with sickle‐cell trait did not differ from comparablel groups of American blacks, both the % Hb F and % F‐cells in Saudi Arabian patients with sickle‐cell anaemia were much higher than in Black. The high Hb F levels in individuals wilth sickle‐cell anaemia are not due to coexistent glucose‐6‐phosphate dehydrogenase defilcency or α‐thalassaemia trait, and the Hb F level showed an inverse correlation with the degree of haemolysis. These findings indicate that the unusually elevated levels of Hb F are not due to an associated high frequency of a gene for hetero‐cellular herditary persistence of fetal haemoglobin in the oasis population, but rather from a genteically determind absolute increase in Hb F production related in some way to the SS genotype
Plasma Levels of Platelet Factor 4 Measured by Radioimmunoassay
Authors: Chesterman C. N. , McGready Jennifer R. , Doyle D. J. , Morgan F. J.
Published: March 2008

Summary. A radioimmunoassay has been developed to measure platelet factor 4 (PF4) in biological fluids both in vitro and in vivo. The assay has been shown to be highly specific for PF4 and has a sensitivity of 0.08 ng/assay tube and 1.6 ng/ml of plasma. The preparation of plasma for the measurement of in vivo levels of PF4 requires the use of an anticoagulant containing EDTA, theophylline and prostaglandin E1, the immediate cooling of the blood and high speed or prolonged centrifugation to reduce platelet contamination. Plasma levels of PF4 are normally between 4 and 24 ng/ml with a median of 7.4 ng/ml.

Plasma PF4 levels are markedly increased during cardiopulmonary bypass surgery with shortened 51Cr‐labelled platelet survival times and during arterial thrombosis. However, despite similarly shortened platelet survival times, the level of PF4 is normal in immune thrombocytopenia. Elevations of plasma PF4 levels are found following surgery, acute myocardial infarction and frequently during acute infections and in inflammatory states. On the contrary, normal levels are usual in disseminated malignancy, in severe hepatic and renal disease and in chronic arterial disease.

The measurement of PF4 is a useful addition to the study of platelet pathophysiology. It is apparent, however, that raised levels are by no means specific for thromboembolic disease and similarly platelet destruction is not invariably associated with abnormally increased plasma PF4 levels.