bluuebunny/crossref_metadata_2025 huggingface dataset:
Ferrokinetics and Erythropoiesis in Man: Red‐Cell Production and Destruction in Normal and Anaemic Subjects
Authors: Cavill I. , Ricketts C. , Napier J. A. F. , Jacobs A.
Published: March 2008

Recent advances in the analysis of plasma 59Fe clearance have produced a unified method for measuring effective and ineffective erythropoiesis (Ricketts et al, 1975). We have used this method to investigate the balance between red‐cell production and destruction in normal subjects and in patients with megaloblastic anaemia, iron deficiency anaemia and refractory hypoplastic anaemia. The results show that the normal marrow can maintain an appropriate red‐cell mass by altering red‐cell production to match destruction. In the anaemias we have studied there is an increased rate of either intra‐ or extra‐medullary red‐cell destruction. The response of the marrow may be limited by iron supply, by defective nuclear maturation or by some intrinsic marrow defect.
HISTOLOGICAL INVESTIGATION OF METHACRYLATE EMBEDDED BONE MARROW BIOPSY SPECIMENS; CORRELATION WITH SURVIVAL AFTER CONVENTIONAL TREATMENT IN 15 ADULT PATIENTS
Authors: te Velde J. , Haak H. L.
Published: March 2008

Aplastic anaemia is a rare disease with a poor prognosis: 40% of the patients die within 6 months after diagnosis. The clinical condition and laboratory data show only a poor correlation with the chance of survival for longer than 6 months. Conventional bone marrow biopsy investigations did not have any predictive value, but methyl‐methacrylate embedded biopsy material from 15 patients given only supportive care showed histological features considered important for the assessment of the prognosis. In 3 μm methacrylate sections non‐myeloid cells, like lymphocytes, could be readily distinguished from erythropoietic cells. Three types of inflammatory infiltrate were easily distinguished according to amount and distribution. Seven of the eight patients with marked infiltration (Grade III) died within 6 months after biopsy, but none of the patients with a sparse infiltrate (Grade I), including five patients in remission. In six of 10 patients, who were not in remission, histological data permitted correction of the clinically estimated prognosis.
Spectrum of von Willebrand's Disease: a Study of 100 Cases
Authors: Mannucci P. M.
Published: March 2008

The clinical picture, family history and laboratory findings of 100 patients with von Willebrand's disease (VWD) have been studied in Italy in a multicentre survey from two rural areas with a known high incidence of the disease and from two large cities (Rome and Milan). Bleeding time, procoagulant factor‐VIII (VIIIAHF) assay, platelet retention and PRP‐ristocetin aggregation were measured in each centre, and plasma samples were frozen and subsequently assayed for factor VIII‐related antigen (VIIIAGN) and ristocetin co‐factor (VIIIVWF) in one laboratory (Milan). On the basis of the inheritance pattern, clinical severity of the disease and laboratory findings, patients with VWD were separated into two groups. In 17 patients the absence of a family history of bleeding, a high incidence of parental consanguinity and involvement of both sexes suggested an autosomal recessive mode of inheritance; the unusual severity of the disease with markedly abnormal results of all six laboratory measurements (and frequent reduced levels of VIIIAGN and VIIIVWF in the unaffected parents) were consistent with the homozygous state. In 83 patients, the disease was familial (being transmitted as an autosomal dominant trait) and of moderate clinical severity. In these, VIIIVWF was usually much lower than VIIIAHF and VIIIAGN. The findings suggest that although some patients may present with simultaneous abnormalities of all six studied laboratory measurements, the majority show a variation in the spectrum of the abnormal results, without any obvious simple pattern.
Congenital Dyserythropoietic Anaemia, Types I and II: Aberrant Pattern of Erythrocyte Membrane Proteins in CDA II, as Revealed by Two‐dimensional Polyacrylamide Gel Electrophoresis
Authors: Anselstetter Volker , Horstmann Hans‐Joachim , Heimpel Hermann
Published: March 2008

The protein composition of the erythrocyte membrane of two patients with CDAI and four patients with CDA II has been investigated by two‐dimensional polyacrylamide gel electrophoresis. The two‐dimensional patterns of erythrocyte membrane proteins of the patients with CDA were compared with the well‐established pattern of normal individuals. No alterations could be detected in the patients with CDA I, but a striking deviation from normal was observed in all patients with CDA II. The major aberrations of the erythrocyte membrane proteins in CDA II are the lack of protein B1.1, the drastically increased concentration of protein B1.4 and the presence of a new component, C2′. These results are discussed in relation to the known morphological and immunological abnormalities of the erythrocyte membrane in CDA II.
Biphasic Thrombopoietic Response to Severe Hypobaric Hypoxia
Authors: Jackson Carl W. , Edwards Carol C.
Published: March 2008

Thrombopoiesis has been studied during and after an 11 d exposure to discontinuous hypobaric hypoxia. Exposure of rats to 0.4 atmospheres for 16‐17 h daily initially caused an increase in platelet count which reached a peak of 1.5 times baseline on days 4 and 5. This thrombocytosis was followed by a decrease in platelets to a nadir of 50‐60% of baseline on days 12 and 13. That thrombocytosis results from increased platelet production is supported by increased [35 S]sulphate incorporation into platelets and increased megakaryocyte size and turnover. The thrombocytopenia with continued hypoxia seems to result from decreased platelet production since 51Cr‐platelet survival was normal while megakaryocyte concentration was decreased to one‐half that of untreated controls. These observations suggest that differentiation of precursors into megakaryocytes was decreased during the thrombocytopenic period, although the fewer remaining megakaryocytes appeared stimulated because of their larger size and increased [3H]thymidine labelling. Thus, hypobaric hypoxia had a biphasic effect on thrombopoiesis with increased platelet production in the first few days of exposure followed by subnormal production.
The Immunotherapy of Acute Myelogenous Leukaemia using Intravenous BCG
Authors: Whittaker J. A. , Slater A. J.
Published: March 2008

In a 2‐year period, 37 of 81 adults with acute myelogenous leukaemia achieved complete remission after repeated courses of Daunorubicin (DNR) and Cytosine Arabinoside (ARAC). They were randomized to maintenance treatment with monthly DNR/ARAC, or to identical chemotherapy plus intravenous BCG. Eighteen BCG treated patients had significantly longer survival times than 19 patients treated with chemotherapy only although no statistically significant difference can be seen in the remission duration of the two groups. Eleven patients in the BCG treated group who have relapsed, have received DNR/ARAC reinduction and five second and two third remissions have been obtained. Twelve control group patients have relapsed and 10 have received further reinduction treatment with DNR/ARAC but only one patient has entered a complete remission. Seven patients in the BCG treated group who survived for 75 weeks or more (76, 76, 96, 124, 125, 138 and 145 weeks) were either PPD positive before treatment or converted to PPD positivity after BCG treatment. Using a battery of skin tests it may be possible to define a good prognostic group of patients and design future treatment accordingly.

The BCG group had a total of 198 intravenous treatments. All patients had pyrexia 6‐12 h after injection lasting 12‐72 h and occasionally headaches and muscle pains. Two patients had non‐fatal anaphylactic reactions which did not recur when BCG was subsequently re‐administered. Other complications of BCG therapy were not a problem and we have not needed to withdraw treatment for any patient.
Inherited Sialoglycoprotein Deficiencies in Human Erythrocytes of Type En(a ‐)
Authors: Anstee D. J. , Barker Diana M. , Judson P. A. , Tanner M. J. A.
Published: March 2008

We have investigated the membranes of erythrocytes from a family in which there is a genetic defect (previously described as the En(a ‐) condition) resulting in the loss of the major erythrocyte sialoglycoprotein (PAS‐1). The results show that two different types of sialoglycoprotein deficiency can be distinguished within this family. We suggest that the En(a ‐) group of variants is more appropriately described as a class of sialoglycoprotein deficient erythrocytes.

Using a new technique it is shown that the blood group M antigen of normal erythrocytes is found only on the erythrocyte sialoglycoprotein while in this family the M antigen is found on membrane components other than the sialoglycoprotein. Our results suggest that the amino acid sequence of the sialoglycoprotein is important in defining the difference between the blood group M and N antigens in normal erythrocytes.
Immunoglobulin Synthesis in Hairy Cell Leukaemia
Authors: Golde David W. , Stevens Ronald H. , Quan Shirley G. , Saxon Andrew
Published: March 2008

In vitro studies were performed with leukaemic cells from two patients with hairy cell leukaemia in order to define the nature and kinetics of immunoglobulin synthesis by the neoplastic cells. Both patients had clinically and morphologically well‐defined disease and their cells contained abundant tartrate‐resistant acid phosphatase. One patient had associated macroglobulinaemia. The hairy cells had B‐lymphocyte characteristics as determined by fluorescent immunoglobulin staining and surface receptor properties. They synthesized monoclonal IgM and IgG respectively in vitro. The kinetics of immunoglobulin synthesis were different in cells from the two patients as measured by equilibration time, intracellular degradation, and secretion. Permanent cell lines were established with cells from these patients. The lines grow as typical B‐lymphoblastoid cultures and continue to produce tartrate‐resistant acid phosphatase and immunoglobulin. These studies unequivocally demonstrate the B‐lymphocyte nature of the hairy cells in these patients and provide evidence for their clonal origin both in terms of immunoglobulin and enzyme synthesis.
Abnormal Folate Polyglutamate Ratios in Untreated Pernicious Anaemia Corrected by Therapy
Authors: Perry Janet , Chanarin I.
Published: March 2008

The relative amounts of folate compounds having 1–3‐, 4–, 5– and 6–glutamic acid residues in the molecule were measured in red blood cells. The results were expressed in relation to methyltetrahydrofolate pentaglutamate (5‐glutamic acid residues) which is the analogue present in highest concentration. When the concentration of pentaglutamate is given a value of 100 the relative concentration of folates with 1–3‐, 4‐, 5‐ and 6‐glutamic acid residues in control subjects are 48, 48, 100 and 58 respectively.

In folate deficiency there is a fall in the relative amount of the short chain glutamic acid chain compounds in addition to having an overall fall in the amount of folate present, the values for 1–3, 4, 5 and 6 chain folates being 16, 14, 100 and 39. By contrast, in untreated pernicious anaemia there is an accumulation of the short chain folates as well as an overall fall in polyglutamate concentration and the relative concentrations were 72, 44, 100 and 30.

Folate polyglutamate appears to be the active coenzyme and the relative amounts of glutamic acid residues may serve to regulate the rate of enzyme activity. In pernicious anaemia the amount of enzyme is reduced and on this hypothesis the regulatory function impaired.
The Conformation of Fibrinogen and Fibrin: An Electron Microscope Study
Authors: Blakey P. R. , Groom M. J. , Turner R. L.
Published: March 2008

A mica adsorption technique was used to prepare fibrinogen and early fibrin polymers for examination in the transmission electron microscope. Globular particles 10.3 nm in diameter were observed and a model for the process of aggregation of these globules into fibrous units is proposed. A comparison is made between the results obtained by the mica adsorption technique and those obtained by conventional preparation techniques involving fixation and staining.
Non‐invasive Screening for Venous Thromboembolic Disease
Authors: Gordon Y. B. , Cooke E. D. , Bowcock Sally A. , Ratky Susan M. , Pilcher M. F. , Chard T.
Published: March 2008

Venous thromboembolism is an important cause of morbidity and mortality, and the clinical signs are notoriously unreliable. Two safe and reliable non‐invasive techniques have therefore been developed for the diagnosis of DVT namely thermography, and the measurement of serum levels of fibrinogen degradation fragment E (FgE). A total of 93 medical patients were studied using these techniques and the results were compared with the phlebographic appearances of the lower limbs. In 91 patients, the thermographic findings were identical to the radiological findings; there was one false‐positive thermograph, and one false‐negative thermograph, which occurred in a case of isolated calf vein thrombosis. There were 59 subjects without evidence of DVT only one of whom had an FgE level greater than 400 μg/l., whereas 17 of the 19 subjects with extensive DVT proximal to the knee, had levels greater than 400 μg/l. In eight subjects with clinical and perfusion lung scan evidence of pulmonary emboli, the FgE levels were all greater than 800 μg/l. These findings demonstrate the reliability of a combination of these two non‐invasive diagnostic techniques in the detection of DVT. The local lesion may be identified using thermography, whereas elevated levels of FgE indicate the likelihood of extensive thrombosis with embolization.
Thromboxane Synthesis and the Platelet Release Reaction in Bernard‐Soulier Syndrome, Thrombasthenia Glanzmann and Hermansky‐Pudlak Syndrome
Authors: Malmsten C. , Kindahl H. , Samuelsson B. , Levy‐toledano S. , Tobelem G. , Caen J. P.
Published: March 2008

Thromboxane (TX) synthesis was investigated in some characterized platelet disorders and correlated with release of ADP or [14C]serotonin and platelet aggregation. In a case with Bernard‐Soulier syndrome TX‐synthesis as well as ADP‐[14C]serotonin release and platelet aggregation were found to be essentially normal when platelets were incubated with collagen, ADP, arachidonic acid or prostaglandin G2 (PGG2). In a case with Hermansky‐Pudlak syndrome aggregation was normal with ADP, whereas aggregation with collagen and PGG2 was markedly decreased. Release of ADP and [14C]serotonin was found to be decreased with all investigated inducers in this platelet disorder, whereas TX‐synthesis was normal with arachidonic acid and PGG2. In three cases with Glanzmann's thrombasthenia almost no aggregation was observed with any of these compounds. However, in this disorder platelet TX‐formation from arachidonic acid or PGG2 was also normal whereas ADP and collagen both induced less TX‐formation than in normal platelets.

These results indicate that none of the investigated platelet disorders was associated with abnormal TX‐synthesis from arachidonic acid as found in platelet cyclo‐oxygenase deficiency. These observations provide further evidence for the hypothesis that the Bernard‐Soulier syndrome and Glanzmann's thrombasthenia are primarily associated with glycoprotein abnormalities at receptor level (Nurden & Caen, 1974, 1975, 1976). A hypothetical model for the interrelationship of two specific glycoproteins and thromboxane‐synthesis is discussed.
Cobalamin Dependent Methionine Synthesis and Methyl‐Folate‐Trap in Human Vitamin B12 Deficiency
Authors: Sauer Hansjörg , Wilmanns Wolfgang
Published: March 2008

The activity of methionine synthetase (MS) is important for the rapid growth of human haematopoietic cells and cultured lymphoblastoid cells. The MS reaction is the only known metabolic step in which both vitamin B12 and folate are essential in a single enzyme reaction. In vitamin B12 deficiency the MS activity in bone marrow cells is significantly lower than that in normal bone marrow.

Free tetrahydrofolic acid (H4PteGlu) is normally liberated from its metabolically inactive storage form, 5‐methyl‐H4PteGlu (CH3H4PteGlu), in the cobalamindependent MS reaction. Thus, in vitamin B12 deficiency H4PteGlu is not available in sufficient concentration to maintain the de novo synthesis of thymidylate and purines, and accords with the methyl‐folate‐trap hypothesis.

After treatment with amethopterin (MethotrexateTM), the incorporation of 3Hdeoxyuridine into cellular DNA is reduced. In proliferating normal cells this effect of methotrexate can be prevented (and the cells rescued) with CH3‐H4PteGlu or with CHO‐H4PteGlu (5‐formyl‐H4PteGlu; LeucovorinTM). On the other hand, in vitamin B12 deficient bone marrow cells this so‐called rescue‐effect could only be achieved with CHO‐H4PteGlu and not with CH3‐H4PteGlu. These observations also support the hypothesis of the methyl‐folate‐trap in vitamin B12 deficiency.

Decreased MS activity in vitamin B12 deficiency seems to be the essential metabolic fault, which is responsible for secondary alterations of folate metabolism. Thus, measurement of MS activity may allow direct functional assessment of vitamin B12 deficiency, at least with regard to DNA metabolism.
Changes in Electrophoretic Mobility of Human Factor VIII‐Related Antigen: Evidence for Subunit Structure
Authors: Ekert Henry , Chavin S. I.
Published: March 2008

Therapeutic factor‐VIII concentrates were found to have factor VIII‐related antigen (FVIIIRAg) with an electrophoretic mobility faster than that of plasma using two‐dimensional crossed immunoelectrophoresis. Immediately after infusion of factor‐VIII concentrates into patients, the electrophoretic mobility of FVIIIRAg in the patients’plasmas was increased to that of the antigen in the infused concentrates. Two hours after infusion, a proportion of the antigen had an electrophoretic mobility intermediate between that of the pre‐infusion antigen and that of the concentrate antigen, and by 24 h after infusion the reversion of electrophoretic mobility to pre‐infusion values was complete. The return of electrophoretic mobility to normal did not occur in vitro after 24 h. In vitro mixing experiments between preinfusion plasma and concentrate resulted in antigen with a range of intermediate mobilities which were related to the relative proportions of slow and fast antigen in the mixture. In vitro mixing experiments with slow and fast antigen separated from intermediate purity factor VIII concentrate by agarose gel filtration resulted in the formation of a relatively large proportion of antigen with intermediate electrophoretic mobility. The most reasonable interpretation of the results is the formation of hybrids between the two electrophoretically different populations of antigen. This implies that VIIIRAg normally exists in a polymeric form which can spontaneously dissociate into and exist in equilibrium with a pool of partially and/or completely dissociated subunits.
Basophilic Chronic Granulocytic Leukaemia with Hyperhistaminaemia
Authors: Rosenthal Susan , Schwartz J. H. , Canellos G. P.
Published: March 2008

Two patients with typical Philadelphia chromosome positive chronic granulocytic leukaemia (CGL) developed an accelerated phase of the disease characterized by an increased white blood cell count and marked basophilia in the bone marrow and peripheral blood. Histamine levels were extremely high in both patients. Hyperhistaminaemia was manifested as wheezing, urticaria, diarrhoea, and pruritus in one patient and as peptic ulcer disease and peripheral oedema in both patients. In one case, gastric acid studies revealed a very high basal to stimulated ratio (BAO/MAO). Treatment with the investigational agent metiamide, an H2 receptor histamine antagonist, resulted in marked improvement in symptoms and reduction in gastric acid output. Extreme basophilia in CGL may be associated with hyper‐histaminaemia, and manifestations of both the H1 and H2 type may occur.
Incorporation of 32PO4 into Phospholipids of Blood Platelets
Authors: Leung N. L. , Kinlough‐rathbone R. L. , Mustard J. F.
Published: March 2008

Human or rabbit platelets in an artificial medium without phosphate were incubated with carrier‐free [32P]orthophosphate for 1 h, washed and re‐suspended in Tyrode‐albumin solution containing unlabelled phosphate. Aliquots of platelet suspension were subjected to lipid extraction at various times and the extracted phospholipids were separated by thin layer chromatography. The specific radioactivities of triphosphoinositide (TPI) and diphosphoinositide (DPI) were highest during the labelling period and then declined rapidly after the platelets were resuspended in the medium containing unlabelled phosphate. The percentage of total phospholipid 32P in TPI and DPI decreased from over 95% at 1 h to less than 50% at 12 h with either rabbit or human platelets. The specific radioactivity of monophosphoinositide (MPI) reached its greatest value 6–8 h after labelling. In the in vitro incubation studies, phosphatidylcholine (PC), phosphatidylserine (PS) and phosphatidylethanolamine (PE) were negligibly labelled in the initial hours but at 12 h (rabbit or human platelets) PC contained more than 30% of the radioactivity. PE and PS contained less than 10% of the total phospholipid‐bound radioactivity at the end of the incubation period. The pattern of incorporation of [32P]orthophosphate into the inositol phospholipids, PC, PS and PE that had been found in the in vitro studies was confirmed by an in vivo study in which rabbit platelets labelled with 32P in vitro were infused into rabbits and harvested after 35 h. These findings indicate that the phosphates of all the phospholipids studied in these experiments turn over in vitro and in vivo but the phosphates of the inositol phospholipids turn over most rapidly.
Impairment by Heparin of Primary Haemostasis and Platelet [14C]5‐Hydroxytryptamine Release
Authors: Heiden D. , Mielke C. H. , Rodvien R.
Published: March 2008

Heparin was administered to 34 normal subjects by intravenous injection (100 u/kg) and the template bleeding time was significantly increased both 10 min and 120 min following injection. Before heparin the bleeding time was 5.3±1.0 min (mean±1 SD); 10 min after injection it was 9.8±5.6 min (P<0.001); and 120 min after injection it was 7.2±3.9 min (P<0.001). Increases in the bleeding time were unrelated to changes in platelet count, and independent of heparin's effect on plasma coagulation. In blood drawn 10 min and 120 min following heparin injection, there was significantly less [14C]5‐HT released from platelet‐rich plasma (PRP) in response to collagen, 0.41 mM epinephrine and 8 μM ADP, although in vitro addition of heparin (0.1 u/ml, 0.5 u/ml and 2.5 u/ml) to baseline PRP of three subjects did not depress [14C]5‐HT release. Our experiments suggest that intravenous administration of a therapeutic dose of heparin can cause a significant reversible impairment of platelet haemostatic properties, possibly by an indirect mechanism.
A to O Bone Marrow Transplantation in Severe Aplastic Anaemia: Dynamics of Blood Group Conversion and Demonstration of Early Dyserythropoiesis in the Engrafted Marrow
Authors: Marmont A. M. , Damasio E. E. , Bacigalupo A. , Giordano D. , Rossi E. , Reali G. , Gay A. , Dagna‐Bricarelli F. , Brema F. , Carella A. M. , Santini G.
Published: March 2008

Summary. A to O bone marrow transplantation was performed in a 25‐year‐old male affected with severe aplastic anaemia, the donor being an HLA compatible brother. Three plasma exchanges had to be performed with an Aminco separator to remove the original and recurring anti‐A isohaemagglutinins. The dynamics of O to A blood group conversion were followed by means of differential agglutination. An early wave of marked dyserythropoiesis was observed in the engrafted marrow. Mild to moderate GvHD was treated successfully with MTX, bolus high dosage 6‐methyprednisolone and, at relapse, with intravenous ALG.
Impaired Neutrophil Function and Myeloperoxidase Deficiency in Myeloid Metaplasia
Authors: El‐Maallem Hilal , Fletcher John
Published: March 2008

Summary. The ability of neutrophils to phagocytose and kill Candida guilliermondii was investigated in 12 patients with myeloid metaplasia (MM). Following ingestion there was a considerable impairment in the ability of MM neutrophils to kill and digest Candida which was not explained by the very mild impairment in phagocytosis. Quantitative myeloperoxidase measurement revealed an overall deficiency of this enzyme in MM neutrophils and a highly significant correlation between low myeloperoxidase levels and impaired candidacidal activity. Neutrophils from patients with myeloid metaplasia show a pattern of defective microbial killing, high alkaline phosphatase activity and low myeloperoxidase activity which is similar to that seen in severe infections and distinct from chronic granulocytic leukaemia.

The cells of one patient with particularly low myeloperoxidase and defective microbial killing were further studied both cytochemically and by electron microscopy. The azurophilic granules of his neutrophils were present in normal numbers and contained normal amounts of acid phosphatase but they lacked myeloperoxidase.
Myelotoxicity of Methotrexate in Animals with Pyogenic Infection
Authors: Harding B. , Mac Lennan I. C. M.
Published: March 2008

Summary. Rats stimulated into prolonged increased neutrophil production by the induction of a unilateral pyo‐hydronephrosis showed consistent profound neutropenia when methotrexate (MTX) was given during the first 2 d of infection. Thereafter greater neutropenia than that observed in non‐infected rats was only observed in occasional animals. There was a significant correlation between the degree of neutropenia induced by MTX and the day after MTX upon which this occurred. The main target for myelotoxicity by MTX appears to be the myelocyte. Its precursors seem relatively insensitive.
Thymoma, Pure Red Cell Aplasia, Pernicious Anaemia and Candidiasis: a Defect in Immunohomeostasis
Authors: Robins‐Browne R. M. , Green R. , And J. Katz , Becker D.
Published: March 2008

Summary. A patient is reported who presented with anaemia and oral candidiasis. Investigation revealed a spindle cell thymoma, and a serum inhibitor of haem synthesis associated with pure red cell aplasia. He was also shown to have the Addisonian pernicious anaemia defect, diagnosed on the basis of a depressed serum level of vitamin B12, 57Co‐cyanocobalamin absorption studies, the presence of intrinsic factor blocking antibodies and atrophic gastritis with an elevated serum gastrin level. A serum inhibitor of candida‐induced lymphocyte transformation, which may have accounted for the chronicity of the candidiasis, was also demonstrated. Following thyniectomy, whereas the antibodies to intrinsic factor and gastric mucosa, and the serum inhibitor of candida‐induced lymphocyte blastogenesis persisted, the inhibitor of haem synthesis disappeared. Moreover, red cell autoagglutinins, associated with a mildly shortened red cell life span, were detected for the first time. Although the patient's anaemia improved following thymectorny, mild megaloblastic changes were noted in the bone marrow. Parenteral vitamin B12 therapy reversed these changes and a normal level of haemoglobin has subsequently been maintained. The candidiasis, however, has persisted. It is suggested that the defect in immunohomeostasis in the patient may have resulted from an abnormality or deficiency of specific suppressor T‐lymphocytes.
The Effect of Endotoxin on Circulating Lymphocytes in Normal Man
Authors: Gale R. P. , And G. Opelz , Golde D. W.
Published: March 2008

Summary. The effect of endotoxin administration on peripheral blood lymphocytes was studied in normal man. A partially purified bacterial endotoxin from Pseudomonas aeruginosa was given intravenously to 10 healthy volunteers. Endotoxin administration resulted in a substantial lymphocytopenia with reduction of both thymus‐dependent T and bone marrow‐derived B lymphocytes. Absolute numbers of circulating B lymphocytes decreased to a greater extent than T lymphocytes. T‐lymphocyte functional characteristics, including mitogenic response to phyto‐haernagglutinin and responder cell activity in mixed lymphocyte culture, were unaffected by endotoxin administration. Response to concanavalin A was consistently reduced. Stimulation of allogeneic lymphocytes in mixed lymphocyte culture, a function of B lymphocytes and monocytes, was also decreased. When endotoxin was used as an in vitro mitogen, a significant increase in DNA synthesis was observed only in lymphocytes from individuals previously given endotoxin and not in normal control lymphocytes. These studies indicate that experimental endotoxaemia is associated with a reduction in absolute numbers of circulating T and B lymphocytes, a decrease in B‐lymphocyte bur not T‐lymphocyte functional activity, and redistribution of a concanavalin A‐responsive subpopulation of T lymphocytes. The data suggest that endotoxin has important effects on lymphocyte distribution and function in man.
A CRM‐Positive Variant of Factor‐VII Deficiency and the Detection of Heterozygotes with the Assay of Factor‐like Antigen
Authors: Mazzucconi M. G. , Mandelli F. , Mariani G. , Briët E. , Veltkamp J. J.
Published: March 2008

Summary. Nine patients with severe factor‐VII deficiency, belonging to seven pedigrees were studied for the presence of factor‐VII‐CRM with an inhibitor neutralization assay. The antibody, raised in rabbits, did not precipitate the antigen and could only be used in a fluid phase assay to measure the capacity of plasma to neutralize inhibitory activity directed against factor‐VII activity. In one of these nine patients normal amounts of factor‐VII‐CRM could be demonstrated. The CRM+ patient did not show a clinical picture at variance with that of the CRM — patients. The investigation into this CRM+ pedigree revealed heterozygosity in nine out of 12 persons when using the ratio between biological factor‐VII activity and factor‐VII‐CRM as the criterion.
Blood Lymphocyte Surface Markers and Clinical Findings in Chronic Lymphoproliferative Disorders
Authors: Jayaswal U. , Roath S. , Hyde R. D. , Chisholm D. M. , Smith J. L.
Published: March 2008

Surface receptors on peripheral blood lymphocytes have been studied in 43 cases with chronic lymphocytic leukaemia (CLL), and five cases with lymphoma and overspill. One quarter of the cases with CLL had lymphocytes with no detectable surface immunoglobulin (SIg) by direct fluorescent antibody staining. The remainder had SIgM, which was associated with SIgD in one third of the cases. 70% of the SIg negative group had extensive extramedullary involvement (EMI) compared with 3 5 % of the SIg positive group at presentation; correspondingly more of the SIg negative group were treated. 46% of the SIg negative group were CLL patients of more than 3 years standing compared with 9% of the SlgM + D group and 32% of the SIgM group. Some explanations for this pattern are discussed.
Drug Uptake and Ribonucleotide Profiles of Blast‐enriched and Blast‐depleted Human Bone Marrow Cell Populations
Authors: Preisler H. D. , Rustum Y. , Walczak IRENE
Published: March 2008

A density cut method was used to prepare two subpopulations of bone marrow cells which were enriched (BE) and depleted (BD) respectively of blast cells. Marrow aspirates were obtained from 32 patients, 26 of whom had acute leukaemia. The uptake of a variety of chemotherapeutic agents by these two subpopulations and the acid‐soluble ribonucleotide profiles of the populations were compared and significant differences were found in drug uptake by BE and BD subpopulations. For some drugs such as cytosine arabinoside, uptake was greatest by the BE cells, while for 5‐azacytidine the BD subpopulation took up the greatest amount of the drug. The preparation of the BE subpopulation also permitted the recognition of several patients with acute leukaemia whose blast cells possess nondetectable levels of ATP, UTP, and GTP components, in their acid‐soluble fractions. The studies presented demonstrate the necessity of using purified cell populations when characterizing the drug uptake patterns and the soluble ribonucleotide profiles of the leukaemic cells. A simple method for enriching bone marrow aspirates for leukaemic cells is also presented.
Comparison of Heparin and a Semi‐synthetic Heparin Analogue, A73025
Authors: Lane D. A. , Michalski R. , Van Ross Maureen E. , Kakkar V. V.
Published: March 2008

A study has been made of a low molecular weight semi‐synthetic heparin analogue, A73025, that may be clinically useful as an antithrombotic agent because of its reported high specificity for antithrombin III. The clearance from the circulation of both heparin and the analogue has been studied in man following intravenous injection. Heparin obeyed almost first order kinetics when assayed using a specific anti‐Xa assay and first order kinetics when measured with KCCT. At high concentrations the heparin analogue was cleared with first order kinetics when assayed both with the anti‐Xa assay and with KCCT. At low concentrations the analogue produced between one half and two‐thirds of the anti‐Xa activity of an equal dose of heparin, producing only a small prolongation of KCCT. With increasing dose, the more specific anti‐Xa potentiating effect of A73025 decreased because of a flattening of its anti‐Xa dose‐response curve. These results suggest that the analogue might be useful as an antithrombotic agent when it is used as a prophylactic agent.
A Sex Difference in the Platelet Count
Authors: Stevens R. F. , Alexander M. K.
Published: March 2008

A study of the platelet counts of 868 blood donors aged 18‐65 showed a higher mean count in women than in men, the difference in the premenopausal age groups being highly significant (P<o.ooI) and of the order of 40‐70 × 109/l. The mean counts of women taking a contraceptive pill did not differ significantly from those who were not.

Values for the normal platelet count in man quoted in most standard haematological texts are derived from studies made on relatively small numbers of individuals using visual counting techniques. The advent of automatic counting and data processing has made practicable the examination and evaluation of larger bodies of data with a relatively higher degree of precision than previously. This communication reports the results of such a study made on blood donors. The work was suggested by the finding of a significantly higher mean platelet count for women in the routine counts made selectively on a hospital population.
High Resolution Autoradiographic Studies of RNA, Protein and DNA Synthesis during Human Eosinophil Granulocytopoiesis: Evidence for the Presence of RNA on or within Eosinophil Granules
Authors: Wickramasinghe S. N. , Hughes Madeleine
Published: March 2008

Human bone marrow cells which had been incubated with [3H]uridine or [3H]leucine for 1 h were studied using the technique of electron microscope‐autoradiography. The autoradiographs revealed the presence of newly‐synthesized RNA and protein molecules within or on a proportion of (1) the primary and secondary granules in all classes of eosinophil precursors and (2) the secondary granules in eosinophil granulocytes. It is suggested that the granule‐associated RNA molecules may be concerned with the synthesis of at least some of the new protein molecules which were incorporated into the limiting membrane or substance of eosinophil granules long after the immature primary granule stage. Studies of eosinophil precursors which had been incubated with [3H]thymidine for 1 h showed that the eosinophil granules do not label with this DNA precursor.
The Measurement of Total and Unsaturated Iron‐Binding Capacity in Serum
Authors:   International Committee for Standardization in Haematology
Published: March 2008

An inter‐laboratory investigation into the assay of serum iron binding capacity by the magnesium carbonate adsorption method was undertaken. The source of MgCO3, storage temperature of whole serum, lyophilization and storage temperature of lyophilized serum were identified as sources of inter‐laboratory variation. Protocols for the clinical assay of UIBC and TIBC are given which have achieved satisfactory inter‐laboratory precision.
Beta‐Thalassaemia Trait: Imprecision of Diagnosis at Birth
Authors: Alter B. P.
Published: March 2008

Globin chain synthesis was studied at birth in infants who had been tested for haemoglobinopathies while in utero. Subsequent haematological studies showed that five infants were normal, five had β‐thalassaemia trait, and two had α‐thalassaemia trait. Correct assignment of these diagnoses could not be made in individual cases from globin chain studies at birth, although the β/α ratios did cluster into groups. By total radioactivity, the mean β/α ratio was 0.38 in the normal infants, and 0.22 in those with β‐thalassaemia trait. Mean (β+γ)/α was 0.90 in the former, and 0.70 in the latter. Specific activity ratios (counts per minute, per absorbance at 280 nm) failed to distinguish thalassaemia trait from normal, as a group or individually. Accurate diagnosis of β‐thalassaemia trait required classical haematological studies during the first year of life.
Capacity of Ringed Sideroblasts to Synthesize Nucleic Acids and Protein in Patients with Primary Acquired Sideroblastic Anaemia
Authors: Wickramasinghe S. N. , Hughes Madeleine
Published: March 2008

Bone marrow cells from two patients with primary acquired sideroblastic anaemia were studied using the technique of electron microscope autoradiography. Whereas a significant proportion of erythroblasts containing small to moderate quantities of intramitochondrial deposits incorporated [3H]thymidine into DNA, the majority of cells containing large quantities of intramitochondrial deposits did not. These results are consistent with the idea that several of the early polychromatic cells with large quantities of deposits become arrested in their progress through the cell cycle. The data also revealed that the accumulation of increasing quantities of iron‐containing material within the mitochondria of an erythroblast was frequently (but not invariably) associated with and was possibly responsible for (1) a depression of RNA synthesis and (2) a more marked depression of protein synthesis. A small proportion of the erythroblasts showing a depression of protein synthesis also displayed various non‐specific ultrastructural abnormalities indicative of cellular injury such as the presence of intranuclear clefts, nuclear membrane‐associated myelin figures and lipid‐laden intracytoplasmic vacuoles.
Further Evidence for the Role of Thrombin in the Platelet Release Reaction Caused by Various Agents, and the Nature of Biphasic Platelet Aggregation
Authors: Huzoor‐Akbar  , Ardlie Neville G.
Published: March 2008

Studies were undertaken to resolve conflicting reports about the possible involvement of thrombin in the release reaction and second phase aggregation caused by ADP.

Washed human platelets suspended in a buffered solution responded poorly to ADP, with no second phase aggregation or release of [3H]5HT. In contrast, washed platelets suspended in dialysed plasma containing all intrinsic pathway clotting factors responded to ADP with second phase aggregation and release of radioactivity. This response depended on an optimum calcium concentration above which aggregation was impaired. Biphasic aggregation and release of platelet contents by ADP, epinephrine and collagen in dialysed plasma was inhibited by hirudin and heparin. The inhibitory effects of hirudin and heparin were not observed when citrate was present.

These observations provide further evidence for the central role of thrombin in the release reaction and biphasic aggregation caused by various agents and indicate that these platelet reactions are not artifacts due to a citrate. Since the effect of citrate on aggregation and the inhibitory actions of hirudin and heparin were produced by another chelating agent, viz EGTA, it is suggested that the citrate effect is due to reduction of cation concentration.
Automated versus Manual Techniques for the Prothrombin Time Test: Results of Proficiency Assessment Studies
Authors: Poller L. , Thomson Jean M. , Yee K. F.
Published: March 2008

Approximately 70% of hospitals participating in the series of prothrombin time proficiency assessment trials organized from the National (U.K.) Reference Laboratory for Anticoagulant Reagents and Control and continue to use a manual technique routinely. The remainder employ a large variety of different types of automated procedure. A comparison has therefore been made of the performance with the recommended manual technique and automated procedures used in the prothrombin time test. There is no indication that automation eliminated local technique variables or reduces the inter‐laboratory error of prothrombin time estimations. Most of these instruments gave shorter prothrombin times than did the manual technique. The practical effect of this difference was reduced by the use of the prothrombin ratio system of reporting but this does not eliminate problems arising from some makes of instrument. The use of a specific correction factor for each make of instrument to compensate for the difference from the manual technique is not recommended. This is because there is too much variation in the performance of individual instruments of the same make. The only practical solution appears to be for all instruments to be calibrated regularly using the recommended procedure of the British System for Anticoagulant Control. The results may be then expressed as British Corrected Ratios.
A Modified Sedimentation Method for Counting Platelets in Blood
Authors: Archer R. K. , Allen B. V. , Baldwin C.
Published: March 2008

A sedimentation method for counting human blood platelets is proposed which uses a fluid of sp.gr. 1.057 at an osmolarity of 320 milliosmols. The fluid is a mixture of Lymphoprep and Isoton II. After centrifugation at about 200 g platelets are counted on an electronic particle counter.
Cell Membrane Enzymes: L‐γ‐Glutamyl Transpeptidase, Leucine Aminopeptidase, Maltase and Trehalase in Normal and Leukaemic Lymphocytes
Authors: Kramers Mary T. C. , Catovsky D.
Published: March 2008

Several cell‐membrane enzymes, which serve functions in amino acid and sugar transport, were measured in peripheral blood lymphocytes from chronic B and T lymphocytic disorders, blast cells in acute leukaemias, and in normal lymphocytes from cord blood, peripheral blood of adults, tonsils and bone‐marrow plasma cells in myelomatosis.

The specific activities of l‐γ‐glutamyl transpeptidase, maltase and trehalase were low, as compared with those measured in normal blood lymphocytes, in the acute leukaemias and in the chronic B‐cell disorders. In myelomatosis and in the chronic T‐cell disorders, the specific activity of these three enzymes was in the normal range or above normal. The specific activity of leucine aminopeptidase was low in all the chronic B‐cell disorders and in some of the lymphoblastic leukaemias. It was elevated in Sézary syndrome cells but low in T‐chronic lymphocytic leukaemia. All four enzymes were lower than normal in cord blood lymphocytes and higher than normal in tonsils. These findings are discussed in relation to the patterns of lymphoid cell differentiation and maturation in normal tissues and in leukaemic states.
Chronic Lymphatic Leukaemia: Cellular Effects of Glucocorticoids in Vitro
Authors: Homo Françoise , Duval Dominique , Meyer Philippe , Belas Fatiha , Debre Patrice , Binet Jacques‐Louis
Published: March 2008

Glucocorticoid receptor levels and steroid induced inhibition of nucleic acid precursors have been examined in lymphocytes from 27 patients at different stages of chronic lymphatic leukaemia. No correlation can be found between the level of glucocorticoid receptors and the stage of the disease. On the other hand, a significant difference (P < 0.02) was found between stage 0 and stage III/IV patients, in terms of the in vitro effect of dexamethasone on [3H]uridine incorporation.
Myelomatosis and the Hyperviscosity Syndrome
Authors: Preston F. E. , Cooke K. B. , Foster Margaret E. , Winfiel D. A. , Lee D.
Published: March 2008

Eleven patients are described in whom myelomatosis was complicated by the laboratory and clinical features of the hyperviscosity syndrome (HVS). The myeloma type was IgA in nine and IgG3 in two. In those patients with IgA myeloma the HVS was related to the presence of high molecular weight complexes in the serum. Remission of clinical features was obtained in all patients by plasma exchange. Clinical improvement coincided with reduction of whole blood viscosity and in those patients with IgA myeloma, with a parallel reduction of the high molecular weight complexes.

The relationship between the IgA complexes and blood viscosity has been examined by physicochemical analysis of purified IgA monomer and polymers and evidence is presented to show that the IgA polymer has a higher intrinsic viscosity and axial ratio than the larger IgM molecule. The significance of these observations is discussed.
Serum Uracil Levels in Acute Childhood Leukaemia
Authors: Parry T. E.
Published: March 2008

Serum ‘uracil + uridine’ expressed as uracil has been measured in 47 consecutive cases of acute leukaemia in children at the time of diagnosis, before treatment, and also both before and after therapy in a girl of 12 with Ph1 positive chronic myeloid leukaemia in blastic transformation, and in an adult male age 31 with acute undifferentiated leukaemia. The results were compared with 97 previously reported normal subjects. The skew distribution of results in acute leukaemia rendered the mean value and standard deviation meaningless but the range was considerably wider than in the normal controls (range 7.2–77.1 μmol/l. in acute leukaemia; mean 15.7 μmol/l., range 5.7–40.5 μmol/l., SD 5.26 in controls). In 10 (21%) of the patients with acute leukaemia the level was more than 3 SDs and in five more than 8 SDs above the normal mean. There was a high degree of correlation between uracil concentration and total white cell count (r= 0.7894, P<0.001) and blast cell count (r= 0.7160, P<0.001) but no correlation was observed between serum uracil and the blood urea or the serum uric acid, vitamin B12 or folate. The grossly elevated uracil levels, white cell and blast cell counts rapidly returned to normal or near normal after therapy.

Retention of uracil by the kidney, increased DNA catabolism and the enzymatic deamination of cytosine are considered, and each in turn excluded, as a possible cause of the observed elevated uracil levels. It is submitted that the raised uracil level is a function of the circulating blast cell and that the results are consistent with the hypothesis that, at least in some cases of acute leukaemia, there may be an impairment in the amination of uracil to form cytosine akin to the impaired methylation of deoxyuridylate in the megaloblastic anaemias. The former would enhance the known mutagenic action of uracil, an action which is absent in the latter. The molecular mechanism, whereby this could give rise to a population of genetically abnormal cells incapable of normal differentiation, is discussed.
Chromosomes in Acute Non‐lymphocytic Leukaemia
Authors:   First International Workshop on Chromosomes in Leukaemia
Published: March 2008

The Workshop analysed clinical‐cytogenetic correlations in adult patients with acute nonlymphocytic leukaemia (ANLL) with special regard to the following questions: (1) Is there an association between the clinical‐haematological diagnosis and a specific chromosome abnormality? (2)Is there a correlation between response to treatment and survival and a particular chromosome abnormality? (3) Is the presence of a chromosomal abnormality or the frequency of abnormal cells (i.e. all metaphases normal (NN), some metaphases normal (AN), no metaphases normal (AA)) predictive of survival or response to therapy? (4) Is there a karyotypic evolution during the course of the disease?