bluuebunny/crossref_metadata_2025 huggingface dataset:
Leukaemia Classification: a Study of the Accuracy of Diagnosis in 456 Patients
Authors: Whittaker J. A. , Withey J. , Powell D.E.B. , Parry T.E. , Khurshid M.
Published: March 2008

Summary. A panel of five haematologists has examined, without consultation or prior knowledge of the diagnosis, blood films and bone marrow smears from 456 patients with a diagnosis of leukaemia. A diagnostic classification which recognized various subtypes of acute myelogenous leukaemia was used but no attempt was made to subdivide acute lymphoblastic leukaemia.

Complete agreement with the initial diagnosis was low (56.4%) and was particularly poor (45.7%) when the patient had one of the forms of acute leukaemia. However, disagreements which would have involved the patient in a change of treatment were unusual (2.0%).

We conclude that a high degree of diagnostic agreement for patients with leukaemia is unlikely from morphological classifications alone.
Stem Cell Migration Induced by Erythropoietin or Haemolytic Anaemia: the Effects of Actinomycin and Endotoxin Contamination of Erythropoietin Preparations
Authors: Quesenberry P. , Levin J. , Zuckerman K. , Rencricca N. , Sullivan R. , Tyler W.
Published: March 2008

Summary. The injection of erythropoietin or the induction of anaemia with phenylhydrazine leads to changes in murine pluripotent and granulocyte‐macrophage stem cells indicating migration from marrow to spleen. In order to evaluate the interrelationship between erythroid differentiation and stem cell migration we have selectively suppressed erythroid differentiation with actinomycin D. Anaemia or EP injection resulted in stem cell changes consistent with migration; actinomycin blocked these changes in anaemic but not EP injected mice while blocking erythropoiesis in both groups.

The erythropoietin contained from 0.01 to 1000 μg/ml of endotoxin as defined by the limulus test; it decreased marrow erythropoiesis and stimulated marrow granulopoiesis. Adsorption of the erythropoietin preparation with limulus lysate removed endotoxin without decreasing erythropoietin activity. Adsorbed erythropoietin stimulated erythropoiesis and not granulopoiesis, and stem cell changes induced by its administration were largely blocked by actinomycin, suggesting that endotoxin in the non‐adsorbed erythropoietin caused the actinomycin resistant stem cell changes.

The observation that actinomycin blocks both erythroid differentiation and stem cell migration suggests that these two physiologic events are closely linked. The effects of injected erythropoietin on murine haemopoietic stem cells may, to a significant extent, be secondary to the presence of endotoxin in the erythropoietin preparations.
The Incidence of Antibodies to Low Frequency Antigens (LFA) in Plasmapheresis Donors with Hyperimmune Rh Antisera
Authors: Contreras M. , Barbolla L. , Lubenko A. , Armitage S. E.
Published: March 2008

Summary. Over 50% of plasmas from plasmapheresis donors hyperimmunized for Rh antibodies were found to contain antibodies to low frequency red cell antigens (LFA). The majority of these plasmas contained antibodies to more than one LFA. On the other hand, the incidence of these antibodies in immune plasma from control plasmapheresis donors was markedly lower. The significance of the high incidence of these antibodies in grouping reagents is discussed.
Specificity in the Factor VIII Response to Vasopressin Infusion in Man
Authors: Prowse C. V. , Sas G. , Gader A. M. A. , Cort J. H. , Cash J. D.
Published: March 2008

Summary. Structural requirements of the systemic factor VIII response to intravenous vasopressin in man has been investigated using vasopressin analogues. With the analogues available the receptor specificity of this phenomenon could not be distinguished from those associated with the previously described plasminogen activator release or antidiuretic effects of this hormone. Further studies using I‐desamino‐[8‐d‐arginine]vasopressin showed a dose‐related release of both procoagulant and antigenic components of the factor VIII complex. The newly released factor VIII could not be distinguished from circulating factor VIII on the basis of molecular size, electrophoretic mobility or in vitro stability despite apparent differences in the duration of response of the procoagulant and antigenic components in vivo.
Some Aspects of the Biology of Multinucleate and Giant Mononucleate Erythroblasts in a Patient with CDA, Type III
Authors: Wickramasinghe S. N. , Goudsmit R.
Published: March 2008

Summary. Marrow cells of a patient with CDA, type III, were (I) incubated with 3H‐thymidine for 0.5 h and studied using a combination of Feulgen microspectro‐photometry and light microscope autoradiography and (2) incubated with 3H‐thymidine, 3H‐uridine or 3H‐leucine for 1 h and studied using the technique of electron microscope autoradiography. The data revealed multiple abnormalities in the proliferation of erythroblasts but no abnormality in the distribution of neutrophil promyelocytes and myelocytes in the different stages of interphase. Some mononucleat erythroblasts had DNA contents of 4–20c and the multinucleate erythroblasts had total DNA contents of 2–40c. In about 40% of the multinucleate erythroblasts, only some of the nuclei within the same cell incorporated 3H‐thymidine and the electron microscope autoradiographic studies revealed that the nuclei which failed to synthesize DNA virtually always showed abnormalities in the electron‐density of the heterochromatin or euchromatin or a Swiss‐cheese appearance of the heterochromatin. Furthermore, in some multincleate cells but not in others, the ultrastructurally abnormal nuclei showed a marked depression of RNA synthesis when compared with the normal‐looking nuclei within the same cell. An occasional binucleate, multinucleate and giant mononucleate erythroblast showed ultrastructural changes suggestive of advanced degeneration and such cells, which showed a marked depression of RNA and protein synthesis, appeared to be phagocytosed by macrophages.
Abnormal Red‐Cell Metabolism of Pyridoxine Associated with β‐Thalassaemia
Authors: Anderson B. B. , Perry G. M. , Modell C. B. , Child J. A. , Mollin D. L.
Published: March 2008

Summary. Red‐cell conversion of pyridoxine to pyridoxal phosphate was studied in control subjects, and patients with heterozygous and homozygous β‐thalassaemia. In 7% of control subjects the rate of pyridoxine conversion was well below the range found in the other control subjects (5.0–8.6%, mean 6.5%/g Hb × 10−2) but in heterozygous β‐thalassaemia was below that range in 63% of the patients. The conversion rate was also slow or borderline in the majority of patients with severe transfusion‐dependent homozygous β‐thalassaemia, in spite of the presence of some donor cells; but was normal, or fast as in other anaemias, in all but one patient with mild homozygous thalassaemia. There was a much higher incidence of a slow conversion rate in the parents of the severe homozygotes than in parents of the mild homozygotes, illustrating the familial pattern. This supports our view that the red‐cell conversion rate of pyridoxine is an inherited characteristic, independent of thalassaemia.

The cause of a reduced rate of pyridoxine conversion was investigated. The increase to a normal rate following riboflavin ingestion suggests a defect in the activity of the flavin mononucleotide (FMN)‐dependent pyridoxine phosphate oxidase.
Studies on the Subcellular Organelles of Neutrophils in Chronic Granulocytic Leukaemia with Special Reference to Alkaline Phosphatase
Authors: Rustin G.J.S. , Peters T.J.
Published: March 2008

Summary. The organelle pathology of neutrophils in chronic granulocytic leukaemia (CGL) was investigated by analytical subcellular fractionation. There were minor reductions in activity of some granule enzymes with an abnormal distribution in sucrose density gradients of the specific granules. There was a marked reduction of 5′‐nucleotidase activity but this is probably related to the relative reduction of the mononuclear cell contamination of the neutrophils isolated from leukaemic patients compared with controls. Another plasma membrane enzyme, NADH‐nitroblue tetrazolium reductase, which has a microbicidal role, had increased activity.

Neutrophils from patients with CGL had 13% the alkaline phosphatase activity of controls and were compared with neutrophils from women in the third trimester of pregnancy when the activity was increased to 8 times the control level. The latent activity, per cent inhibition by Levamisole, kinetic constants and subcellular distribution of alkaline phosphatase were similar in the three groups. It is suggested that the properties and intracellular localization of alkaline phosphatase are normal in CGL and that there is a quantitative lack of enzyme.
Synthesis of Procoagulant Factor VIII, Factor VIII Related Antigen and other Coagulation Factors by the Isolated Perfused Rat Liver
Authors: Shaw E. , Giddings J. C. , Peake I. R. , Bloom A. L.
Published: March 2008

Summary. The synthesis of factor VIII and other coagulation factors has been studied using an isolated, perfused rat liver. Synthetic function of the liver was validated by adding [35S]l‐methionine to the perfusion medium and performing two‐dimensional crossed immunoelectrophoresis and autoradiography on samples obtained during perfusion. Progressive incorporation of radioactivity into plasma proteins was demonstrated. This was inhibited by cycloheximide. Coagulation factor assays demonstrated synthesis of factors II, IX and X and of factor V and procoagulant factor VIII (VIIIC). Synthesis of factor VIII related antigen (VIIIRAg), measured in an immunoradiometric assay, was not significantly demonstrated. Addition of warfarin to the perfusion medium inhibited the synthesis of factors II, IX and X but not of factors V and VIII. Cycloheximide completely inhibited synthesis of all coagulation factors but actinomycin acted only after a latent period. Reticuloendothelial cell blockade was attempted by adding ethionine to the perfusion medium or by administration of Indian ink to the donor animals prior to removal of the livers. In these instances synthesis of factor V and factor VIIIC was inhibited but not that of factors II, IX and X. The results confirmed the functional capacity of the isolated liver for synthesizing proteins and the vitamin K dependent coagulation factors, and suggested similar kinetic features for the synthesis of factors V and VIIIC. Failure to detect significant VIIIRAg synthesis in these experiments is consistent with the hypothesis that this protein is released by vascular endothelial cells throughout the body and is activated or joined to VIIIC or stimulates its production in the liver.
Serum Inhibitors of Myelopoiesis
Authors: Fitchen John H. , Cline Martin J.
Published: March 2008

Summary.

A modification of the microcytotoxicity assay of Terasaki & McClelland was used to screen for serum inhibitors of myeloid progenitor cells (CFU‐C). Sera from 104 patients with neutropenia or bone marrow failure and from 104 controls, including 30 normal subjects and 74 patients with various disorders or multiple transfusions, were studied. Inhibitors of CFU‐C were found in 19 of the 104 patients with neutropenia or marrow failure: three with acquired neutropenia and 16 with aplastic anaemia. In 17 of the 19 patients the inhibitor was either complement dependent or characterized as an immunoglobulin. Manipulations aimed at reducing the titre of inhibitor were convincingly associated with clinical remission of disease in one patient. In another patient, the inhibitor was an IgG antibody directed against non‐HLA antigens present on CFU‐C of all 50 normal target marrows tested; however, it was not clearly autoreactive. Inhibitors were also found in one of nine patients with lupus erythematosus, one with breast carcinoma, one patient with renal transplant rejection, and six of 46 multiply transfused patients without marrow failure. Our findings indicate that most serum inhibitors are associated with multiple transfusion: convincing evidence of pathogenetically important autoreactive antibody was present in only one of 19 patients with inhibitors.
Structure‐Function of the Factor VIII Complex Studied with an Immobilized Heteroantisera to VIII:C
Authors: Cooper Herbert A. , Lee Dorothy , Lamb Mary Ann , Wagner Robert H.
Published: March 2008

Summary.

An antibody was raised in rabbits to the small active fragment of human factor VIII, obtained by Ca2+ dissociation of a human factor VIII preparation made from a multidonor plasma pool. After absorption, the antibody neutralized the factor VIII coagulant activity of normal human plasma, but did not precipitate with any plasma or plasma fractions or neutralize von Willebrand factor (vWF) activity as measured by ristocetin aggregation of fixed washed platelets. Immune beads were prepared by CNBr binding of the partially purified rabbit antibody to 1% agarose beads. Non‐immune beads were prepared with IgG fractions obtained from the rabbits before immunization and used throughout as a control. The amount of factor VIII coagulant activity (VIII:C) removed from plasma by immune beads was time‐dependent and proportional to the amount of beads used, but all of the VIII:C could not be readily removed. Removal of VIII:C by immune beads parallelled removal of factor VIII:antigen, but less vWF activity was removed. Immune beads could be blocked or saturated by treatment with large amounts of normal plasma, but not by von Willebrand disease plasma and only by some haemophilic plasmas.
Heparin and Partial Thromboplastin Time: an International Survey
Authors: Poller L. , Thomson J. M. , Yee K. F.
Published: March 2008

Summary.

The reliability of routine partial thromboplastin time (PTT) methods in the measurement of the anticoagulant effect of heparin has been assessed in a study involving over 300 hospitals in the U.K. and overseas. Commercial PTT methods were relatively insensitive to heparin, added in vitro, compared with the standardized PTT method tested by the same laboratories. Non‐commercial, locally‐prepared reagents compared well with the standardized method particularly in the detection of a low concentration of heparin. The value of a sensitive reference preparation for the calibration of routine PTT reagents used in heparin control is demonstrated.
Globin Chain Biosynthesis in Iron Deficiency
Authors: Walford Diana M. , Deacon Rosemary
Published: March 2008

Summary.

Globin chain synthesis was studied in seven severely iron‐deficient patients before and after treatment with iron. There was no appreciable difference between the individual pre‐ and post‐treatment α/β specific activity ratios and the mean α/β ratio for each group was 1·00±SD 0·04. In a further six untreated iron‐deficient patients the mean α/β ratio was 1·00±SD 0·04. There was therefore no evidence that iron deficiency caused a reduction in α/β ratio.

Three patients with β thalassaemia trait and coexistent iron deficiency had lower α/β ratios before treatment than after treatment with iron. It appeared that iron deficiency had caused reduced α chain synthesis in this group.

Preliminary experiments have shown that the α/β specific activity ratio of purified haemoglobin A is decreased in iron deficiency, indicating an increase in the size of the free α chain pool. It is suggested that iron deficiency may interfere with the proteolytic mechanism normally responsible for the destruction of excess α chains. In combined iron deficiency and β thalassaemia trait, the resulting increase in free α chains might act by negative feedback to inhibit further α chain synthesis (Blum et al, 1970) thereby reducing the pre‐treatment α/β ratio.
Characterization of Null Cells in Chronic Lymphocytic Leukaemia with B‐Cell Allo‐ and Hetero‐antisera
Authors: Kirov Sylvia M. , Kwant W. O. , Fernandez L. A. , MacSween J. M. , Langley G. R.
Published: March 2008

Summary.

Chronic lymphocytic leukaemia peripheral blood mononuclear cells (CLL‐PBMN) were separated into B, T and Null‐enriched lymphocyte sub‐populations using sequential mouse and sheep red blood cell rosetting depletions on Hypaque–Ficoll gradients. The procedure produced viable cell populations with mean percentage purities of 90, 87 and 75 for B, T and non‐rosetting (Null‐enriched) sub‐populations, respectively. More than 80% of PBMN cells were generally accounted for by mouse and sheep rosetting.

The purified lymphocyte sub‐populations were examined with a panel of B‐cell specific alloantisera obtained from kidney transplant recipients and a rabbit antiserum to B cell antigen isolated from a human B‐lymphoblastoid line. The results illustrated that the antigens detected by these sera also have potential as a marker for characterizing the CLL population. Where conventional markers were weak or absent, B cell antigens were readily detected in both fluorescent and cytotoxic tests.

The majority of the non‐rosetting cells (> 90%) in CLL followed similar patterns of reactivity to the purified B cells, suggesting they are a subset of B cells. A small residual population (0–5% of PBMN) did not react with the antisera, the significance of which is unknown.
Kinetics, Distribution and Sites of Destruction of 111Indium‐labelled Human Platelets
Authors: Heyns A. duP. , Lötter M. G. , Badenhorst P. N. , van Reenen O. R. , Pieters H. , Minnaar P. C. , Retief F. P.
Published: March 2008

Summary.

The survival, tissue distribution and fate of 111In‐oxine labelled autologous platelets in six normal humans were studied with serial blood sampling, scintillation camera and computer‐assisted imaging, whole body profile scanning, and rectilinear scanning. 111In‐platelets recovery in the circulation was 72±16% and survival was 216±17 h. Platelet survival curves fitted a linear function best. Initially platelets pooled rapidly in the spleen as a single exponential function, and at 90 min 26% of the injected 111In was located in this organ. Early hepatic uptake was also significant and at 90 min constituted 16% of total body 111In‐activity. As labelled platelets disappeared from the circulation there was a threefold increase of radioactivity in the liver to reach 39% of whole body activity at 216 h. Radioactivity also increased significantly in the spleen (33±3% at 216 h). There was significant residual radioactivity in the thoracic and lower abdominal regions at 216 h, suggesting that platelets are also sequestered in the bone marrow. Radioactivity in the lower limbs almost disappeared with time (0±7% at 216 h), indicating that utilization of platelets in the peripheral vasculature is not marked in normal subjects.
Increase in Coagulation Factor VIII Activity in Man following Acute Hypoglycaemia: Mediation via an Adrenergic Mechanism
Authors: Corrall R. J. M. , Webber R. G. , Frier B. M.
Published: March 2008

Summary.

Coagulation factor VIII activity has been assayed in nine normal volunteers before and after acute hypoglycaemia. In all subjects there was a rise in activity with a mean increment of 96%. Control studies using saline failed to demonstrate any change. In four subjects with pre‐ganglionic sympathectomy no increase in activity was observed. In the normal subjects the increase was blocked by propranolol but not by metoprolol, indicating mediation via beta2 receptors.
Urinary β‐Thromboglobulin in Essential Hypertension
Authors: Anderton J. L. , Fananapazir L. , Dawes Joan
Published: March 2008

Summary.

Urinary concentrations of the platelet‐specific protein β‐thromboglobulin (β‐TG) were measured in patients with essential hypertension. Values obtained for patients with normal renal function fell within the same population as those for a control group of normal individuals, but nevertheless 21% of these patients had urinary β‐TG values above the upper limit of the normal range. Values for patients with renal insufficiency were significantly different from the other groups, and 44% of these individuals had abnormally elevated urinary β‐TG concentrations. These elevated values may reflect an increased plasma concentration of β‐TG in patients with renal impairment or they could be an indicator of renal damage.
Chronic Myeloid Leukaemia: Evidence for Basophil Differentiation and Histamine Synthesis from Cultured Peripheral Blood Cells
Authors: Denburg J. A. , Wilson W. E. C. , Goodacre R. , Bienenstock J.
Published: March 2008

Summary. We have attempted to assess basophil differentiation in vitro in 15 patients with chronic myeloid leukaemia (CML). Using a sensitive radioassay, whole blood histamine values were found to be elevated in 10 of 14 patients tested, and correlated well with peripheral blood basophil counts (r=+0.70). In seven of nine CML patients, but not in controls, total histamine content of separated peripheral blood cells suspended in a modified Marbrook system was shown to rise after 7 d in vitro. Further study showed that both histamine (mean, five‐fold increase) and basophils (mean, three‐fold increase) were significantly elevated over control values at 1,2 and 3 weeks in vitro. Total cell‐associated histamine content (in pg per 100 viable cells) was greater in CML cultures than in controls at 3 weeks (P<0.01). Serial cultures of cells from one patient revealed substantial in vitro rises in basophils and histamine at an accelerated, but not at a chronic, phase of disease. Cells from this patient and two others studied at the time of blast crisis demonstrated higher indices of basophil and histamine increases when compared to the group of CML patients in chronic phases of the disease (P<0.05). We conclude that basophil precursors exist in increased numbers in the peripheral blood of CML patients. Assays for basophil differentiation may prove useful in following disease activity in this and other myeloproliferative disorders.
Enhancement of the Growth of Blood Monocytes in Agar Cultures by Haemolysates
Authors: Chen Dien‐ming , Lin Hsiu‐san
Published: March 2008

Summary. Mouse blood monocytes grow well in liquid culture but poorly in agar medium. In this study the effect of haemolysate on the clonal growth of blood monocytes in agar cultures was investigated. The results suggested that haemolysate from mouse or rat red blood cells contains a factor(s) that can render the murine blood monocytes able to grow well in agar culture (anchorage‐independent growth). The mechanism of action of the enhancing factor(s) is discussed.
Immunosuppression and Serious Infections in Children with Acute Lymphoblastic Leukaemia:a Comparison of Three Chemotherapy Regimes
Authors: Rapson N. T. , Cornbleet M. A. , Chessells J. M. , Bennett A.J. , Hardisty R. M.
Published: March 2008

Summary. Eighty‐four children presenting with acute lymphoblastic leukaemia were entered into a trial designed to test the effect on host toxicity of regular drug‐free periods during chemotherapy. Patients received the same total dose of drugs either continuously (daily), intermittently (a 5 d course every 3 weeks) or in an intermediate way between these two (a 14 d course followed by a 7 d gap).

Mean neutrophil counts were lower in the intermittent group and fell significantly at 6 week intervals, after courses which included prednisolone and vincristine in addition to methotrexate and 6‐mercaptopurine. Mean lymphocyte counts, mitotic response to phytohaemagglutinin and plasma immunoglobulin levels were significantly lower in the continuous group. The results in the intermediate group fell between those of the other two groups. All six remission deaths occurred in the 42 patients in the continuous group, who had a much higher incidence of infections (mostly viral and protozoal) than the other two groups. It is concluded that the intermittent chemotherapy schedule permits the maintenance of a lymphocyte population size and function which provides a satisfactory level of defence against infection without prejudice to its anti‐leukaemic effect.
Glucose‐6‐Phosphate Dehydrogenase Deficiency and Homozygous Sickle Cell Disease in Jamaica
Authors: Gibbs W. N. , Wardle Jane , Serjeant G. R.
Published: March 2008

Summary. The relationship between D‐glucose‐6‐phosphate: NADP oxido‐reduc‐tase (E.C.I.1.1.49; glucose‐6‐phosphate dehydrogenase; G6PD) deficiency and homozygous sickle cell (SS) disease was examined in 120 patients. The proportion of hemizygotes (22‐6%) was slightly more than that observed, and the combined proportions of heterozygotes and homozygotes (28‐3%) were slightly less than would be expected, in the general population, but the differences were not significant. However, the proportion of patients of abnormal G6PD status in the 10‐19 years age group was 41‐7%, significantly more than that found in the 20‐29 years age group (0‐02<P<0‐05), or expected in the general population (P=0‐05). Possible reasons for this are discussed. Difference in G6PD status did not affect the total haemoglobin concentration, reticulocyte count, unconjugated serum bilirubin or Hb F concentration, irreversibly sickled cell counts or plasma haemoglobin concentration, and there was no demonstrable correlation between clinical severity or leg ulceration and abnormal G6PD status.
A Factor VII Concentrate for Therapeutic Use
Authors: Dike G. W. R. , Griffiths D. , Bidwell E. , Snape T.J. , Rizza C. R.
Published: March 2008

Summary. A concentrate of factor VII suitable for therapeutic use has been prepared from human plasma by a method forming part of a comprehensive scheme of large‐scale plasma fractionation. Factor VIII was separated as cryoprecipitate and factors II, IX and X were adsorbed on DEAE‐cellulose. Most of the factor VII remained in the supernatant. By batch adsorption on DEAE‐Sepharose, followed by elution on a chromatographic column, factor VII was concentrated about 25‐fold, and purified about 50‐fold compared with original plasma, without the need for further dialysis or concentration steps. Data are presented from 10 batches, each from 80‐120 kg plasma. Following doses of factor VII to six congenitally deficient patients, the mean rise in plasma factor VII was 95–100% of theoretical; the half‐disappearance time was about 4 h. The treatment of four patients with acquired deficiency of factor VII is also described. No untoward side effects were observed.
Contribution of the Platelet Factor V Content to Platelet Factor 3 Activity
Authors: van Zutphen H. , Bevers E. M. , Hemker H. C. , Zwaal R. F. A.
Published: March 2008

Summary. The procoagulant activity obtained from bovine thrombocytes has been compared to that of lipids isolated from platelets, with and without the addition of purified bovine factor V. A one‐stage assay, which consisted of delipidated bovine plasma containing RVV‐activated factor X, was used to assess the activity. At low lipid concentrations no difference in coagulant activity was found between sonicated vesicles of extracted platelet lipid and lysed platelets. At higher lipid concentrations, however, the extracted lipids were found to be less active than lysed platelets. Determination of factor V in suspensions of gel‐filtered platelets demonstrated that suspensions containing 2 × 109 platelets per ml possessed about 1% of the factor V activity present in a normal bovine plasma pool. Platelet lysis by sonication produced a five‐fold increase in factor V activity. Addition of factor V to sonicated vesicles of extracted platelet lipid, so as to produce an identical factor V activity per amount of lipid as found in lysed platelets, decreased the clotting time only in the higher lipid concentration range. A further three‐fold increase in the amount of factor V added to the lipid vesicles made the coagulant properties of the lipid vesicles indistinguishable from those of lysed platelets over the whole range of phospholipid concentrations tested. When the conditions of the test were changed by diminishing the concentration of factor Xa in the substrate plasma, the difference between lysed platelets and extracted platelet lipid disappeared completely. It is concluded that the higher coagulant activity of lysed platelets, as compared to that of extracted platelet lipid, can be ascribed to platelet factor V activity. Therefore there is no compelling necessity to postulate the existence of a specific procoagulant factor in the platelet other than factor V or phospholipids.
The Role of Colony Stimulating Activity in Modulating Murine Diffusion Chamber Granulopoiesis
Authors: Sullivan Richard , Zuckerman Kenneth S. , Quesenberry Peter J.
Published: March 2008

Summary

. We studied the effects of high circulating Colony Stimulating Activity (CSA) levels and irradiation induced marrow hypoplasia in CF1 and C57B1/6J host mice upon granulopoiesis in intraperitoneal diffusion chamber (DC) cultures. Serial endotoxin injections resulted in marked elevation of circulating CSA for the first half of an 8 d culture period, and CSA was shown to diffuse into the chamber environment; yet this manipulation alone did not significantly accelerate DC cell growth. Pre‐irradiation of the host mice produced no elevation of circulating CSA during the early phase of culture, but resulted in significant stimulation of DC granulopoiesis. Fluctuations in circulating inhibitors of in vitro granulopoiesis did not correlate well with DC cellularity. We conclude that endogenous CSA elevation does not provide an effective stimulus per se for granulocyte‐monocyte proliferation within DC culture and cannot be solely responsible for mediating the exuberant DC granulopoietic response seen in the pre‐irradiated host.
Prolymphocytic Leukaemia: an Ultrastructural Study of 22 Cases
Authors: Costello Christine , Catovsky D. , O'Brien Maureen , Galton D. A. G.
Published: March 2008

Summary

. Twenty‐two cases of prolymphocytic leukaemia (PLL) have been studied by transmission electron microscopy (TEM); 17 had B‐cell surface markers (B‐PLL) and five had T‐cell characteristics (T‐PLL). The predominant cell, the prolymphocyte, has distinct features which were common to all cases: it is a relatively large lymphoid cell with a prominent nucleolus, well condensed peripheral nuclear chromatin and a variable amount of heterochromatin in intranuclear clumps.

Eight cases of PLL (seven B‐PLL, one T‐PLL) showed minor morphological deviations from typical PLL which may present diagnostic difficulties when studied only at light microscopy: the cells in four of these had some features in common with those of chronic lymphocytic leukaemia (CLL), namely round regular nuclei and heavy chromatin condensation, but the nucleoli were larger in the PLL cases. In four other cases nuclear clefting was a prominent feature at TEM.

B‐PLL can be distinguished by ultrastructural features from other B‐cell malignancies, e.g. follicular lymphoma, B‐CLL and B‐acute lymphoblastic leukaemia.
Cytogenetic Follow‐up Study of Acute Non‐lymphocytic Leukaemia
Authors: Lawler Sylvia D. , Summersgill Brenda , MacD. Clink H. , McElwain T. J.
Published: March 2008

Summary

. A cytogenetic study was made at diagnosis in 68 patients with acute non‐lymphocytic leukaemia. The median survival time of the group of patients was 47 weeks. Patients with totally abnormal karyotypes in the bone marrow had a median survival of 14 weeks, whereas for those with only chromosomally normal cells the median survival was 52 weeks, and 47 weeks for those with normal and abnormal cells. Survival was not influenced by the presence or absence of clonal abnormalities. A cytogenetic follow‐up study was made on 41 of these patients. One of the three patients, still in first remission after 3 years, originally had an 8;21 translocation. In general, the abnormalities present at diagnosis disappeared during remission and there was no evidence that clones of abnormal cells were produced by treatment. The chromosomal findings in relapse were not necessarily the same as those seen at diagnosis and the abnormal karyotypes found were usually not clonal in type. A simple kinetic technique, in which the yield of mitoses after 2 and 24 h is compared, may be used as an adjunct to cytogenetic studies in predicting relapse.
Characterization of a Variant Prothrombin in a Patient Congenitally Deficient in Factors II, VII, IX and X
Authors: Johnson C. A. , Chung K. S. , McGrath K. M. , Bean P. E. , Roberts H. R.
Published: March 2008

Summary. An 18‐month‐old child, who had no evidence of liver disease, malabsorption, or chronic ingestion of coumarin compounds, was found to have plasma deficiencies of factors II, VII, IX and X. Assays for factor II and X by immunological techniques (antibody neutralization and immunoelectrophoresis) revealed normal or elevated antigenic activity of these factors, suggesting the presence of abnormal protein variants in the patient's plasma. On two‐dimensional immunoelectrophoresis of the patient's plasma in calcium, a normal and an abnormal population of prothrombin were seen. The abnormal prothrombin had a mobility more anodal than that of normal prothrombin, but less anodal than that of acarboxyprothrombin. The abnormal prothrombin, in contrast to acarboxyprothrombin, adsorbed readily to both aluminum hydroxide and barium citrate, and could be identified by two‐dimensional immunoelectrophoresis of a barium citrate eluate. We suspect that the abnormal variant represents a partially carboxylated prothrombin.
Sternberg's Lymphoma: Effect of Treatment Regimen upon Prognosis in 38 Cases
Authors: Starke I. D. , Wiltshaw Eve
Published: March 2008

Summary.

Thirty‐eight cases of diffuse poorly differentiated lymphoma with an anterior mediastinal mass were reviewed. Nine patients had stage I disease, 11 stage 11, one stage 111 and 17 stage IV. Survival was prolonged in those stage III and IV cases whose treatment had commenced with chemotherapy rather than with radiotherapy. Survival and relapse‐free interval were prolonged in all cases when chemotherapy had begun within a week of diagnosis and had continued with intervals of not more than 21 d between courses.

Although patients whose chemotherapy had been delayed while radiotherapy was given fared worse overall, the incidence of mediastinal relapse was significantly increased when Mediastinal irradiation had been omitted altogether or had been given in fractions of less than 100 rads. Prophylactic cranial irradiation and intrathecal Chemotherapy reduced the incidence of CNS involvement in all stages of the disease. Testicular involvement occurred in three patients, two of them detected only at post‐mortem, and occult ovarian involvement was also found in two post‐mortems.

Therapeutic principles are proposed including immediate systemic chemotherapy followed by mediastinal irradiation and CNS prophylaxis as soon as practicable after chemotherapy is begun and, later, testicular biopsy, or CT scan or ultrasound assessment of pelvic organs.
Cimetidine and Granulopoiesis: Bone Marrow Culture Studies in Normal Man and Patients with Cimetidine‐associated Neutropenia
Authors: Fitchen J. H. , Koeefler H. P.
Published: March 2008

Summary We studied the effect of the H2‐receptor antagonists, cimetidine and metiamide, on in vitro myeloid colony formation by bone marrow cells from seven normal volunteers and two patients with a cimetidine‐associated neutropenia. A cimetidine concentration of 500 μg/ml produced 50% inhibition of normal granulocyte—macrophage colony formation, and 1000 μg/ml of cimetidine completely suppressed proliferation. The inhibitory effect of metiamide occurred at lower concentrations: 50% inhibition at 250 μg/ml and 95% inhibition at 350 μg/ml. Cimetidine had a similar inhibitory effect on colony formation by recovery marrow from the two patients with cimetidine‐associated neutropenia. Treatment of autologous and allogeneic marrows with patients’ acute phase sera and cimetidine failed to show evidence of antibody‐mediated suppression of granulopoiesis. Our results indicate that at sufficiently high concentrations, cimetidine and metiamide inhibit human bone marrow myeloid colony formation in vitro. These findings are consistent with the hypothesis that H2‐receptor antagonists may produce neutropenia in a dose‐related fashion by injury to granulocytic progenitor cells in vivo.
Evidcnce for the Functional Heterogeneity of the Two Sites of Transferrin in Vitro
Authors: Baarlen Joop van , Brouwer Johannes T. , Leibman Adela , Aisen Philip
Published: March 2008

Summary.

A recently developed crossed immunoelectrophoretic method for displaying and quantitating the four possible molecular species of transferrin has been utilized to assess the relative effectiveness of each site of rabbit and human diferric transferrin in providing iron to rabbit reticulocytes. The site which appears to reside in the N‐terminal half of the rabbit protein was found to be at least 5 times more effective than its counterpart. However, both sites may serve as iron donors in monoferric as well as diferric rabbit transferrins. It is also possible that iron may be removed from rabbit transferrin in pairwise as well as sequential fashion. In human diferric transferrin, the site in the C‐terminal domain functions as the better iron donor for rabbit reticulocytes.
Endothelial Cell Hybrids and the Suppression of Factor VIII Related Antigcn Exprcssion
Authors: Edgell Cora‐Jean S. , Reisner HowardM. , Graham John B.
Published: March 2008

Summary.

Hybrid somatic cell clones have been generated by fusing human vascular endothelial cells in primary culture to cells of four rodent lines. Factor VIII related antigen (VIIIR:Ag) was clearly demonstrable in the cultured endothelial cells, even when they had been co‐cultured with rodent cells. Hut in none of 14 hybrid clones was VII1R; Ag detectable. Isozyme analyses for human chromosome markers show that all the assayed human chromosomes were represented among the hybrids, and that various subsets of human chromosomes have been deleted from individual hybrid clones. It may be concluded, therefore, either that VIIIR:Ag production depends on a particular combination of human chromosomes not represented in any of the hybrids, or that the rodent cells contribute some agent which intracellularly blocks VIIIR:Ag expression.
INCIDENCE OF INFECTIONS IN PATIENTS WITH PELGER‐HUET LEUCOCYTE ANOMALY
Authors: Vaya A. , Aznar J.
Published: March 2008

The Pelger‐Huet (PH) leucocyte anomaly is manifested by the presence in the peripheral blood of granulocytes with a non‐lobulated nucleus, but it is generally assumed that this has no clinical effects. However, by careful observation of a group of 85 subjects (from 32 families) with the PH anomaly the incidence of infections, particularly of local nature, appeared to be higher in these patients than in a control group composed of 85 healthy subjects of the same sex and age, observed for a similar period (Table 1).
Cell Volume Studies in B‐Cell Leukaemia
Authors: Costello Christine , Wardle Jane , Catovsky D. , Lewis S. M.
Published: March 2008

Summary Leucocyte volume histograms were obtained in 12 cases of B‐chronic lymphocytic leukaemia (B‐CLL), two cases of B‐prolymphocytic leukaemia (B‐PLL) and two cases of hairy‐cell leukaemia (HCL) by means of a Coulter model ZB1 linked to a channelyser. Visual examination of the histograms showed differences between the cases which were confirmed by measurement of the modal volume (MV) and the σ value (SD of log‐volume). The MV of the cells in B‐CLL ranged from 153·8 to 244·2 fl (mean 198·24 fl) and was lower than the MV in the cases of HCL (427·5 and 465·6 fl). One case of B‐PLL had cells with an MV within the B‐CLL range while in the other the MV was greater (317·9 fl). The histogram in one of two cases of B‐CLL in ‘prolymphocytoid’ transformation showed a relatively high σ value, thus suggesting heterogeneity in cell size. The volume histogram is a simple method of sizing the predominant cell in the peripheral blood in lymphoproliferative disorders and permits an objective comparison between cell sizes in different diseases.
Separation of Normal Mature Bone Marrow Plasma Cells
Authors: Mendelow B. , Grobicki D. , Katz J. , Metz J.
Published: March 2008

Summary A three‐phase purification process for the separation of normal bone marrow plasma cells is described. Material was obtained from haematologically normal humans and baboons. Known monoclonal and polyclonal B cell activators were avoided both in vivo and in vitro. In Phase I, bone marrow fragments were separated from cell suspensions by buoyant density methods. Marrow fragments were shown to be richer in plasma cells than the corresponding marrow cell suspensions. Phase II consisted of culturing fragments by a simple suspension method in which a selective affinity of plasma cells and marrow stromal cells resulted in further concentration of plasma cells with discharge of haemopoietic elements. Maximum concentration of plasma cells occurred within 7 d. In phase III, fragments were disaggregated with trypsin, and the marrow stromal cells were removed by their adherence properties. The resulting non‐adherent fraction comprised approximately 85% plasma cells. The process allows for direct quantitative evaluation of normal plasma cell physiology in vitro.
Evaluation of a New Chromogenic Assay for Factor VII and its Application in Patients on Oral Anticoagulant Treatment
Authors: Avvisati G. , ten Cate J. W. , van Wijk E. M. , Kahlé L. H. , Mariani G.
Published: March 2008

Summary A new chromogenic method has been developed and rigorously standardized for the estimation of factor VII in defibrinated diluted plasma. This method employs a mixture of CaCl2‐rabbit brain thromboplastin as activator, diluted factor VII deficient plasma as source of factor X and the chromogenic substrate S2222 for the measurement of factor Xa. The chromogenic method was insensitive to cold‐and kaolin‐induced activation of factor VII, this in contrast to the one‐stage clotting assay. Results obtained with the chromogenic method revealed good correlation with the clotting method in 33 normal subjects, in 42 patients on oral anticoagulant therapy and in five patients with severe congenital factor VII deficiency. A good correlation was also obtained with ‘Thrombotest’. Comparative estimation of factor VII and of factor VII cross‐reacting material in supernatants of BaCl2 adsorbed plasma of coumarin treated patients revealed that the chromogenic method does not measure decarboxy factor VII. Detailed investigations revealed a half life for decarboxy factor VII of 2·1±0·6 h.
Functional Restoration of ACD‐Blood by Pyridoxal 5′‐Phosphate
Authors: Maeda Nobuji , Kon Kazunori , Sekiya Misuzu , Shiga Takeshi
Published: March 2008

Summary Successful application of pyridoxal 5′‐phosphate (PLP) to restore the oxygen transport function of ACD‐stored blood is described. PLP is readily incorporated into ACD‐erythrocytes by both carrier‐mediated transport (in which ATP may participate) and passive diffusion. Plasma proteins (up to 2·5%) and inorganic phosphate (up to 40 mm) do not affect the incorporation of PLP, though more than 25 mm inorganic phosphate is necessary for the maintenance of ATP levels. Increasing the PLP concentration and/or decreasing the packed cell volume in the medium, increases the incorporation of PLP. Incubation of erythrocytes with PLP at pH 7·0 is most suitable for incorporation of PLP and the maintenance of ATP levels. PLP incorporated into erythrocytes restores the oxygen transport function of the ACD‐erythrocytes, though decreased haem‐haem interaction is observed. A procedure for the clinical application of PLP‐loaded erythrocytes is suggested.
Erythrocyte Pyrimidine 5′‐Nucleotidase
Authors: Torrance J. D. , Whittaker D. , Jenkins T.
Published: March 2008

Summary In this study 31 family members of a patient with erythrocyte pyrimidine 5′‐nucleotidase deficiency were studied. The activity of this enzyme in their erythrocytes is compared with levels in normal subjects and the problems surrounding heterozygote detection are discussed. The mean erythrocyte pyrimidine 5′‐nucleotidase activity in 158 normal fresh blood samples was 130·SD 29·8 mU/gHb. There was no significant difference between males and females. The enzyme level in a patient with non‐spherocytic haemolytic anaemia was 6 mU/gHb. Of 31 relatives of the enzyme deficient patient examined five were clearly heterozygous for the enzyme defect. Their enzyme levels were below 50 mU/gHb. The father, who is an obligatory heterozygote, had an enzyme level of 87 mU/gHb which falls within the low values of the normal range, the distribution of enzyme activity in 26 family members having enzyme activities greater than 50 mU/gHb suggests that six of these may be carriers of the defective gene. We were unable to identify carriers by enzyme kinetic studies, electrophoresis, chromatographic examination of acid extractable nucleotides or measurement of enzyme levels in young and old erythrocyte populations. The last‐mentioned technique showed that erythrocyte 5′‐nucleotidase activity in reticulocytes may be as high as 1785 mU/gHb and declines rapidly as the cell ages reaching about 50 mU/gHb in the oldest cells. Blood samples which had been stored frozen were examined to see whether such samples were satisfactory for population studies. The mean enzyme activity in 151 blood samples stored frozen for more than 12 months was 153·SD 44·7 mU/gHb. The increase in enzyme levels in the frozen samples appears to be greatest in samples showing haemolysis. In spite of the increased enzyme level frozen samples could be used to detect subjects with enzyme deficiency and some heterozygotes.
Cell Surface and Enzyme Markers of Cord Blood Lymphocytes
Authors: Foa R. , Catovsky D. , Cherchi Maria , Benavides I. , Ganeshaguru K. , Hoffbrand A. V.
Published: March 2008

Summary. Human cord blood (CB) lymphocytes were studied with several markers for T‐ and B‐cells and the results compared with those of adult peripheral blood (PB) samples. The proportion of E‐rosettes was significantly lower in CB (mean 24.7±13.5 SD) than in PB (67.5 ± 7.3 SD). Treatment with neuramidase produced a marked increase in the proportion of E‐rosettes in CB (mean 47±13.9 SD), still below the PB values. The proportion of CB lymphocytes showing block positivity with α‐naphthyl‐acetate‐esterase correlated closely with the percentage of E‐rosettes in neuraminidase treated cells. The percentage of H‐rosettes (human RBC) was significantly higher in CB (7.2±6.0) than in PB (3.2±1.6 SD). Re‐rosetting experiments showed that in CB about 30% of the E‐positive cells formed H‐rosettes, in contrast to 5% in PB. These findings indicate that in CB the real number of T‐lymphocytes is higher than shown by conventional E‐rosette formation.

The proportion of B‐lymphocytes, tested by surface immunoglobulins and by rosette formation with mouse RBC (M‐rosettes), was similar in CB and in adult PB. A slight increase in cells with IgM on the surface was found in CB. The overall proportion of lymphocytes with negative B and T markers in CB is three times greater than in adult PB. Levels of the enzyme terminal deoxynucleotidyl transferase were marginally increased in CB; in two out of 41 samples the levels were above those found in normal bone marrow. CB may be a suitable model for the study of lymphocyte subsets with negative B and T markers in man.