bluuebunny/crossref_metadata_2025 huggingface dataset:
Hairy‐Cell Leukaemia: a B‐Lymphocytic Disorder
Authors: Jansen J. , Schuit H. R. E. , Van Zwet Th. L. , Meijer C. J. L. M. , Hijmans W.
Published: March 2008

Fifteen cases of histologically proven hairy‐cell leukaemia (HCL) were studied with immunofluorescence, rosette, and phagocytosis techniques. Unfixed hairy cells (HC) bound all kinds of labelled antiserum; but after fixation with formaldehyde a much more selective binding was observed. In two cases no surface‐bound Ig was detected; four cases showed γ and in nine cases two or three heavy chains were found, α and δ being the most frequent. Few cases were clearly positive for μ. The picture was invariably monoclonal with respect to light chains. Cytoplasmic Ig was present in only 3/15 cases; it was always IgM.

HC did not form E‐rosettes or react with a fluorescent anti‐T cell antiserum. No EAIgMC‐rosettes were formed. All cases showed Fc receptors, which were detected with EAIgG‐rosettes (13/13) or with antigen–antibody complexes (6/6). The density of Fc receptors varied widely. Incubation with latex particles resulted in cell‐associated particles in 16–63% of the HC; with Staphylococcus epidermidis, the percentage was 2–36. After enzyme treatment (lysostaphin), however, no ingested bacteria were found, which suggests that HC are essentially non‐phagocytic. At least 13 cases were therefore classified as B‐cell malignancies.
Relevance of Colony Forming Assays to Bone Marrow Transplantation with Particular Reference to Grafting for Aplastic Anaemia
Authors: Gordon M. Y. ,   for The Royal Marsden Hospital, Bone Marrow Transplantation Team
Published: March 2008

The value of bone marrow colony‐forming assays in monitoring transplanted patients has been assessed by comparing results from two pairs of bone marrow recipients. One pair received marrow from their identical twins for acute lymphoblastic leukaemia; the other pair were grafted with allogeneic marrow from their siblings for aplastic anaemia. One of each pair showed successful engraftment while in the others the grafts failed.

The colony‐forming assay was then used to investigate marrow function in five grafted aplastic patients. Of these, four rejected their first grafts and required further immunosuppression before engraftment could be accomplished. The remaining patient was immunosuppressed at the outset with antithymocyte globulin (ATG) and her first graft was successful. Sera from all five patients inhibited colony formation by normal human marrow and it is suggested that this activity was related to graft rejection as well as to the pathogenesis of the condition.
Haem Synthesis in Sideroblastic Anaemia
Authors: Konopka L. , Hoffbrand A. V.
Published: March 2008

Measurement of delta‐aminolaevulinic acid (ALA) synthetase has been performed in bone marrow of 24 normal subjects and 20 patients with sideroblastic anaemia (five congenital, two pyridoxine‐responsive, 12 primary acquired and one secondary form due to isoniazid therapy). ALA‐synthetase activity without added pyridoxal‐phosphate in vitro was decreased in four of the congenital cases, and in all 12 primary acquired cases. In two of these 16 cases with low ALA‐synthetase activity and in a case with sideroblastic change secondary to isoniazid therapy, the enzyme activity was corrected to normal by pyridoxal‐phosphate in vitro.

ALA‐synthetase activity was normal in both cases of pyridoxine‐responsive anaemia while they were receiving pyridoxine therapy but in one of these cases, studied at repeated intervals after withdrawal of pyridoxine therapy, the bone marrow ALA‐synthetase activity fell to very low levels unless pyridoxal‐phosphate was added in vitro. In this case the Km of the enzyme for pyridoxal‐phosphate was substantially greater than that of a control sample. Haem synthetase activity was reduced in the bone marrow of two of six cases of primary acquired sideroblastic anaemia and in one congenital case tested but was normal in the pyridoxine‐responsive patient studied on pyridoxine therapy. Nevertheless, this patient showed reduced incorporation of 14C‐labelled delta‐aminolaevulinic acid into bone marrow haem.

It is concluded that both congenital and primary acquired sideroblastic anaemia are a heterogenous group of disorders, with different defects in haem synthesis, reduced activity of delta‐aminolaevulinic acid synthetase being common but not invariable in both types. Pyridoxine‐responsive (congenital) sideroblastic anaemia may be due to the presence of an abnormal ALA‐synthetase apoenzyme which requires excessive amounts of the coenzyme, pyridoxal‐phosphate, to achieve normal activity; other abnormalities of haem synthesis may also be present.
Blocking and Binding Type Antibodies against All Major Vitamin B12‐Binders in a Pernicious Anaemia Serum
Authors: Marcoullis George , Parmentier Yves , Nicolas Jean‐Pierre
Published: March 2008

Summary. The simultaneous occurrence of blocking and binding antibodies to intrinsic factor (IF), transcobalamin (TC‐ II, TC I) and other R type vitamin B12 proteins in the serum of a patient with treated pernicious anaemia (PA) is reported here for the first time. The dialysed and purified immunoglobulin‐G (IgG), but not the immunoglobulin‐M (IgM), from a PA patient neutralized the total unsaturated vitamin B12 binding capacity (UB12BC) of human gastric juice, saliva and serum and also of rabbit serum, suggesting that the PA IgG contained blocking antibodies against, IF, TC II, TC I and other R‐binders. In addition, the PA IgG and IgM preparations contained binding antibodies since they could form macromolecular complexes with 57Co‐B12 bound to IF, TC I or TC II so that each one of the latter was totally excluded from Sephadex G‐200. The presence of the binding antibodies was further confirmed by the formation of radioactive precipitation lines on agarose with each one of the vitamin B12‐binders bound to 58Co‐B12. The PA serum did not exhibit any measurable UB12BC after dialysis against 7.5 m guanidine‐HCl followed by renaturation with phosphate buffer (pH 7.4). But it did form TC I and TC II complexes with 57Co‐B12 when the latter was added during the renaturation step indicating that the serum contained circulating immunoglobulin‐TC complexes. The blocking antibodies should be distinguished from the previously described binding antibodies. The blocking of the binding of vitamin B12 to TCs resulted in relatively lower serum vitamin B12 levels in the present case in contrast to the presence of binding antibodies where high serum vitamin B12 levels have been reported. In addition, the binding antibodies form immunocomplexes with TCs which can easily be detected because they can bind radioactive vitamin B12 while the corresponding immunocomplexes of blocking antibodies are hidden because they prevent the binding of the vitamin to TCs.
β‐Thromboglobulin, Platelet Production Time and Platelet Function in Vascular Disease
Authors: Cella G. , Zahavi J. , De Haas H. A. , Kakkar V. V.
Published: March 2008

Summary. Plasma β‐thromboglobulin (βTG) levels were measured in 103 healthy controls and 112 patients suffering from either peripheral vascular disease (PVD), or cerebrovascular disease (CVD) or deep vein thrombosis (DVT). Plasma βTG was significantly elevated in 46 PVD patients and 24 recent DVT patients compared to controls, but did not differ significantly in 18 chronic DVT and 24 old CVD patients. In addition, heparin neutralizing activity (HNA) and platelet aggregation induced by adenosine diphosphate, 1‐epinephrine and thrombin were compared in 33 out of the 46 PVD patients to 33 controls. The mean HNA was significantly shorter in the PVD patients than in controls. The rate and extent of platelet aggregation were increased in PVD patients compared to controls, but the difference was not statistically significant. Platelet production time (PPT) was measured in 20 controls, 35 PVD patients, nine chronic DVT and 12 chronic CVD patients; significantly shorter PPT was only observed in 14 patients with advanced PVD compared to controls, suggesting increased platelet consumption in these patients. All four assays (plasma βTG, HNA, platelet aggregation and PPT) were performed in 25 patients; no correlation between the four tests was found in these patients suggesting that the tests were measuring various aspects of platelet function. These results suggest that in vivo platelet consumption as well as platelet aggregation and ‘release reaction’are presumably enhanced in PVD and recent DVT patients and that plasma βTG and PPT assays may be better and more specific indicators of in vivo platelet activation than in vitro platelet aggregation test.
Impaired Microtubule Assembly and Polymorphonuclear Leucocyte Function in the Chediak‐Higashi Syndrome Correctable by Ascorbic Acid
Authors: Boxer L. A. , Albertini D. F. , Baehner R. L. , Oliver J. M.
Published: March 2008

Summary. It was previously shown that the abnormal surface characteristics and defective bactericidal function of polymorphonuclear leucocytes (PMN) in the Chediak‐Higashi syndrome (CHS) are correlated with impaired microtubule assembly, and in one patient direct electron microscopic evidence for an anomaly in microtubule assembly following surface membrane activation by concanavalin A (Con A). We show here that very few microtubules are visible in CHS leucocytes from two additional patients under conditions where normal PMNs contain abundant microtubules, and that both in vivo and in vitro exposure of the CHS leucocytes to ascorbic acid promotes the assembly of microtubules. This agent, which normalizes chemotaxis and degranulation in CHS leucocytes, is shown also to correct granulocyte adherence in these leucocytes. It is suggested that the improved clinical course of patients with CHS following treatment with ascorbic acid is related at least in part to improvement of microtubule assembly and PMN function by the ascorbic acid.
Reduced Leucocyte Alkaline Phosphatase Activity and Decreased NBT Reduction Test in Induced Iron Deficiency Anaemia in Rabbits
Authors: Celada Antonio , Herreros Vincente , Pugin Paul , Rudolf Hans
Published: March 2008

Summary. Iron deficiency anaemia was induced in rabbits by repeated bleeding. The leucocyte alkaline phosphatase (LAP) of 26 ± 28 units was significantly reduced compared with control values of 233 ± 36 units (P < 0.001). Leucocyte NBT reduction was also diminished, both in Hanks solution (P < 0.01) and in autologous serum (P < 0.001). After administration of iron, these values returned to normal.

The results suggest that reduced LAP may reflect a deficiency of iron dependent constituents which are necessary for the integrity of normal granulocyte metabolism.
Occurrence of Gγ Hb F in Greek HPFH: Analysis of Heterozygotes and Compound Heterozygotes with β Thalassaemia
Authors: Clegg J. B. , Metaxatou‐Mavromati A. , Kattamis C. , Sofroniadou K. , Wood W. G. , Weatherall D. J.
Published: March 2008

Summary. Haemoglobin F has been isolated from the red cells of individuals with the Greek form of hereditary persistence of fetal haemoglobin (HPFH), and the glycine/alanine composition of the γCB3 peptides determined. In contrast to previous reports we have shown that the Hb F of the Greek HPFH heterozygotes contains significant amounts of Gγ chains and circumstantial evidence indicates that these are the products of the same chromosome that carries the Greek HPFH determinant. Hence this chromosome must be directing the synthesis of Gγ, Aγ and (probably) β and δ chains, thus implying that the Greek form of HPFH does not result from a deletion involving the globin chain structural genes. Analysis of the levels and structure of Hb F from the Greek HPFH heterozygotes and from separated cell populations from the Greek HPFH/β thalassaemia compound heterozygotes indicate that the Greek HPFH determinant, while allowing an overall increase in γ chain synthesis, is not the sole factor determining the absolute amount of Hb F production on a cellular basis.
Non‐excretory Multiple Myeloma
Authors: Joyner Miles V. , Cassuto Jill‐Patrice , Dujardin Pierre , Schneider Maurice , Ziegler Gerard , Euller Liana , Masseyeff Rene
Published: March 2008

Summary. The clinical and laboratory features in five patients with non‐excretory myeloma are described including plasma cell immunofluorescence and, in two cases, ultrastructure. Findings are compared with those in similar patients previously reported, and those in excretory disease. Clinical, haematological and biochemical features were similar to those found in excretory myeloma, showing differences only in relation to the absence of serum or urinary monoclonal immunoglobin. Cellular cytological and ultrastructural features allowed no differentiation from excretory cells. Within this non‐excretory group distinction between secretory and non‐secretory myeloma cells is possible on the basis of immunofluorescence. Differing patterns of intermittent excretion occurred in three of these patients.
Identification of a Pure Splenic Form of Chronic Lymphocytic Leukaemia
Authors: Dighiero G. , Charron D. , Debre P. , Porrier M. Le , Vaugier G. , Follezou J. Y. , Degos L. , Jacquillat Cl. , Binet J.‐L.
Published: March 2008

Summary. We have recently proposed a new staging system for chronic lymphocytic leukaemia (CLL) in which patients with isolated splenomegaly are classified into a distinct stage (stage II). Twenty‐three such patients (from two institutions) have been studied without recorded death in a follow‐up of 18 months to 30 years. This favourable prognosis justifies separation of these ‘pure splenic forms’ (SCLL) which must be distinguished from what Galton has termed prolymphocytic leukaemia (PL). This distinction can be made on the basis of three criteria: (i) Clinically, SCLL has a slow uneventful course and neither anaemia and/or thrombocytopenia; (ii) cytologically PL can be distinguished from other forms of CLL though atypical forms of CLL may be confused with the former; and (iii) the study of surface membrane immunoglobulins (SmIg) showed that while lymphocytes from most patients with both PL and SCLL bore uniform SmIg, suggesting a monoclonal B‐cell proliferation, there was a major quantitative difference in that whereas PL lymphocytes had a number of antigenic sites close to that of normal lymphocytes (mean: 82 000 sites per cell), SCLL lymphocytes had a drastically reduced number of sites. It is our opinion that this is an important criterion for the differential diagnosis between PL and SCLL.
Protection by Ascorbate against Acetylphenylhydrazine‐Induced Heinz Body Formation in Glucose‐6‐Phosphate Dehydrogenase Deficient Erythrocytes
Authors: Winterbourn C. C.
Published: March 2008

Summary. Ascorbate was found to inhibit oxidation of oxyhaemoglobin and Heinz body formation in glucose‐6‐phosphate dehydrogenase deficient red cells incubated with acetylphenylhydrazine. It is proposed that ascorbate can substitute for the glutathione which is depleted and act as a scavenger for drug free radicals generated during the reaction. Ascorbate was protective at concentrations only a little higher than that in normal blood, and the possibility that administration of ascorbate could protect GSH deficient cells against the action of oxidative drugs such as APH is considered. A simple method of quantitatively assessing Heinz body formation by measuring the turbidity of the lysed cells is described.
Clinical Course in 28 Unselected Patients with Aplastic Anaemia Treated with Anabolic Steroids
Authors: Van Hengstum M. , Steenbergen J. , Haanen C.
Published: March 2008

Summary. Twenty‐eight unselected patients with histologically proven aplastic anaemia were electively treated with anabolic steroids (75–150 mg orally q.d.) Additional supportive treatment with blood cell components and antibiotics was given if indicated. Response to therapy was defined as favourable if after 3 months of anabolic therapy overt bleeding tendency had disappeared, there was no need for transfusion therapy, a spontaneous increase of haemoglobin and occurred of > 3 g/dl above the initial level, and a platelet rise of twofold the initial count (up to at least > 30 × 109/1) had occurred. Of 22 patients evaluable for the results of long‐term (>3 months) anabolic treatment, six showed a partial response and eleven responded favourably. These 11 are all alive at the end of the study. Five of these patients proved to be anabolic steroid‐dependent. The 50% actuarial survival is approximately 4 years after diagnosis, which compares favourably with the best published results from bone marrow transplantation for aplastic anaemia. It is concluded that anabolic therapy in aplastic anaemia should be tried for 2‐3 months before the bone marrow transplantation or immunosuppressive therapy is taken into consideration.
Vitamin B6 Metabolism in Anaemic and Alcoholic Man
Authors: Solomon L. R. , Hillman R. S.
Published: March 2008

Summary. Physiological and pathological factors affecting intracellular red cell vitamin B6 metabolism in normal, anaemic and alcoholic man were studied using a new assay for pyridoxine kinase (PnK) together with saturated and total aspartate aminotransferase (EGOT) activities as indirect indices of intracellular pyridoxal 5‐phosphate (PLP) availability. In studies of anaemic states, subjects with iron deficiency anaemia demonstrated elevated levels of both PnK and saturated EGOT, while seven out of 17 subjects with inflammatory anaemia had subnormal PnK but variable saturated EGOT activities. Despite a high incidence of complicating inflammatory disease, alcoholic subjects with or without ring sideroblastic anaemia had elevated levels of both PnK and saturated EGOT. As judged from the saturated EGOT and the ratio unsaturated EGOT/saturated EGOT, intracellular PLP availability was always appropriate to the higher levels of PnK activity.
Liposome Entrapped Desferrioxamine and Iron Transporting Ionophores: a New Approach to Iron Chelation Therapy
Authors: Young S. P. , Baker E. , Huehns E. R.
Published: March 2008

Summary. Liposome‐entrapped desferrioxamine was administered to iron‐overloaded 59Fe labelled mice. When given orally or intraperitoneally entrapment did not enhance the effect of the chelator, but given intravenously liposomal desferrioxamine doubled the 59Fe excretion for a given dose of the drug, and excretion after a single dose continued for up to 3 d. In addition, liposomes containing ionophore A23187 administered concurrently with DTPA caused an excretion of 59Fe, whereas DTPA alone had no effect.
Work Capacity, Heart Rate and Blood Lactate Responses to Iron Treatment
Authors: Ohira Y. , Edgerton V. R. , Gardner G. W. , Senewiratne B. , Barnard R. J. , Simpson D. R.
Published: March 2008

Summary. Changes in haemoglobin (Hb), work performance, heart rate and postexercise blood lactate were studied in iron deficient, anaemic subjects for 16 d following total dosage infusion of iron dextran, i.v. (30–50 ml). Six adult men and 14 women were subjects with initial Hb levels of 6.6±0.6 g/dl (mean±SEM) for the iron treatment group (n=10) and 8.0±0.7 for the placebo group (saline infusion, n=10). Serum iron levels were 0.51±0.15 and 0.67±0.12 mg/l for the two groups, respectively. Haemoglobin and maximal work time increased significantly within 4 d after iron treatment and continued to increase up to 16 d. No changes were found in the placebo subjects. Heart rates at a given exercise intensity were lower in the iron treatment group than in control subjects who had the same Hb levels but had not been treated with iron. Post‐exercise venous blood lactate was similar on succeeding days after iron treatement even though the subjects reached higher work loads.

These results demonstrate that treatment of iron deficient, anaemic subjects with iron dextran results in improved work capacity within 4d and a lower heart rate at a given work load after treatment which cannot be accounted for totally by the elevation of Hb concentration.
Relation between Low Erythrocyte Acetylcholinesterase Activity and Membrane Lipids in Paroxysmal Noctural Haemoglobinuria*
Authors: Siriwittayakorn Junjaree , Yuthavong Yongyuth
Published: March 2008

Summary. Acetylcholinesterase of intact erythrocytes, their ghosts and salt soluble extracts obtained from patients with paroxysmal nocturnal haemoglobinuria (PNH) does not differ from normal with respect to Km values for acetylthiocholine, and Ki values for phenyltrimethylammonium iodide. However, the enzyme from PNH sources has lower Vmax values than normal, has different thermal stability from normal, has less distinctive transition temperature in the Arrhenius plots, and is less subject to inhibition by stearic acid. These results and that from comparison of activation of deoxycholate‐extracted enzyme by lipids from normal erythrocytes suggest that the low acetylcholinesterase activity in PNH erythrocytes is due, at least in part, to alteration in the lipid environment of the enzyme.
In Vitro Culture of Erythroid Colonies from Human Fetal Liver and Umbilical Cord Blood*
Authors: Hassan M. W. , Lutton J. D. , Levere R. D. , Rieder R. F. , Cederqvist L. L.
Published: March 2008

Summary. Colony formation by erythroid precursors from human fetal liver, umbilical cord blood and adult peripheral blood has been studied in a plasma clot culture system. Fetal liver (FL) was obtained at post‐mortem examination from 13–22 week abortuses. After mincing in Hanks' solution, cells in suspension were harvested by Ficoll‐Hypaque centrifugation. Mononuclear cells were obtained by centrifugation of umbilical cord blood (CB) and normal adult peripheral blood (PB). All three types of preparations were incubated up to 14 d in 0.1 ml plasma clot cultures containing 0–4 u/ml erythropoietin (Epo) and 106 cells/ml. No colonies formed in the absence of Epo. Normal adult PB produced late‐appearing colonies; there were no colonies at day 7 and up to 100 colonies/0.1 ml at day 14. CB produced early and late colonies with up to 200 colonies/0.1 ml at day 7 and 125 at day 14. Cells from FL produced many early colonies; over 1500 colonies/0.1 ml were seen at day 7 and there was a subsequent decline in colony count with longer incubation. In cultures of both CB and FL, colonies composed of either mature or immature cells were noted during both early and late stages of incubation suggesting that these cell sources contain a heterogeneous population of erythroid colony progenitors. Measurement of differential β and γ globin chain synthesis by erythroid colonies grown from fetal liver and umbilical cord blood gave results similar to those obtained by direct pulse‐labelling of the original source of the cultured cells.
Congenital Erythrocytosis: a New Form Associated with an Erythropoietin‐dependent Mechanism
Authors: Whitcomb W. H. , Peschle C. , Moore M. , Nitschke R. , Adamson J. W.
Published: March 2008

Summary.

Three patients with probable congenital erythrocytosis were studied to determine the role of erythropoietin (ESF) in their disease. In addition, haemoglobin function was measured and ESF excretion determined in response to reduction in the haemoglobin concentration. In two cases ESF excretion was clearly elevated above normal, and in the third excretion was normal even at an elevated PCV. In all, phlebotomy of 15–20% of the blood volume led to more than doubling in ESF excretion. This occurred in the presence of normal molecular haemoglobin function. Studies of the renal vasculature were normal. These results suggest a new form of congenital erythrocytosis associated with increased ESF production which is reciprocally related to the oxygen carrying capacity of the blood, but in the absence of any demonstrable abnormality in oxygen transport. These findings suggest an inherited defect, likely residing in the renal sensor responsible for the production of ESF. This defect is possibly due to impaired recognition of available oxygen or else a decrease in oxygen supply at the subcellular level.
Myeloid and Erythroid Colony Growth in Non‐anaemic Patients with Fanconi's Anaemia
Authors: Daneshbod‐Skibba Ghodsi , Martin Jean , Shahidi Nasrollah T.
Published: March 2008

Summary.

Myeloid and erythroid colony growth of the bone marrow and peripheral blood cells from one anaemic and four non‐anaemic patients with Fanconi's anaemia were studied. Markedly decreased myeloid and erythroid colony growth was found in all five patients regardless of their haematological status. The decreased colony formation in non‐anaemic patients with Fanconi's anaemia is thought to be due to an intrinsic cellular defect.
Superoxide Dismutase, Glutathione Peroxidase, Catalase and Lipid Peroxidation of Normal and Sickled Erythrocytes
Authors: Das Salil K. , Nair Rajagopalan C.
Published: March 2008

Summary.

Sickled erythrocytes showed reduced glutathione peroxidase and catalase activities in comparison to normal erythrocytes. In addition, increased levels of superoxide dismutase and ‘peroxidation potential’as well as fluorescent lipid pigments and malonaldehyde suggestive of membrane lipid peroxidation were found in sickled erythrocytes. Finally, sickled erythrocytes showed membrane ‘bound’Heinz bodies as well as reduced membrane lipids and unsaturated fatty acids. From these observations, it is suggested that membrane lipid peroxidation occurs in sickled cells.
Shear Rate Dependent Inhibition of Platelet Adhesion and Aggregation on Collagenous Surfaces by Antibodies to Human Factor VIII/von Willebrand Factor
Authors: Baumgartner H. R. , Tschopp T. B. , Meyer Dominique
Published: March 2008

Summary.

The effect of heterologous and homologous antibodies to factor VIII/von Willebrand factor (F.VIII/VWF) and purified VWF on the interaction of human platelets with subendothelium and with a surface consisting of collagen fibrils was investigated using annular flow chambers. Wall shear rates of 200–5200 s−1 were produced by varying the flow rate of the citrated blood which was circulated through these chambers by a pump. Surface coverage with platelets and with platelet aggregates was measured morphometrically. The antibodies had no effect on platelet adhesion at low shear rates. However, an increasing inhibition of adhesion was observed with increasing shear rates. The antibodies to F.VIII/VWF and to purified VWF virtually abolished adhesion at shear rates which are present in the microvasculature (1300–5200 s−1). By contrast, antibodies to factor VIII:C which were isolated from two patients with haemophilia A had no effect on adhesion. In addition antibodies to F.VIII/VWF and to purified VWF inhibited adhesion‐induced aggregation. We conclude that F.VIII/VWF plays an essential role as cofactor for platelet adhesion to subendothelium and collagen fibrils at high blood shear rates, i.e. when the time available for the establishment of a stable bond between platelets and collagenous surfaces is short.
Renal and Extrarenal Erythropoietin Production in Anaemic Rats
Authors: Erslev A. J. , Caro J. , Kansu E. , Silver R.
Published: March 2008

Summary. Rats were rendered anaemic by a single bleeding or by a single injection of phenylhydrazine. At various times after the onset of anaemia they were nephrec‐tomized and challenged with a 6 h exposure to hypoxia. The erythropoietin titre observed at the end of this hypoxic period was corrected for renal erythropoietin induced by the anaemia alone, and the resulting extrarenal component was compared to total erythropoietin production of nephric rats in response to anaemia plus 6 h hypoxia. Extrarenal erythropoietin production was found to increase from 10.3% in normal rats to 12.5% in moderately anaemic rats to 15.1% in rats with severe bleeding anaemia. In phenylhydrazine‐treated rats this extrarenal component was found to be 18.3% possibly due to stimulation of extrarenal erythropoietin by haemolysed red cells. Chronic phenylhydrazine administration resulted in splenomegaly and Kupffer cell hyperactivity but not in any further stimulation of extrarenal erythropoietin production.
Familial Association of the Lupus Anticoagulant
Authors: Exner T. , Barber S. , Kronenberg H. , Rickard K. A.
Published: March 2008

Summary. A lupus anticoagulant was detected in plasma from two pairs of siblings using the kaolin clotting time mixing test. Strong evidence for this inhibitor in one further sibling pair is presented. Although coagulation abnormalities are usually classified as either acquired or of genetic origin it is apparent that the lupus anticoagulant might often be an acquired coagulation defect requiring genetic predisposition.
Red Cell Pyrimidine 5′‐Nucleotidase and Glutathione in Myeloproliferative and Lymphoproliferative Disorders
Authors: Lieberman J. E. , Gordon‐Smith E. C.
Published: March 2008

Summary

. Red cell reduced glutatione (GSH) and pyrimidine 5′‐nucleotidase (Pyr 5′‐NT) were measured in a variety of myeloproliferative and lymphoproliferative disorders. Raised levels of GSH were found in chronic lymphocytic leukaemia, Hodgkin's disease, non‐Hodgkin's lymphoma, Waldenstrom's macroglobulinaemia and myeloma. Decreased activity of Pyr 5′‐NT was found in acute myeloblastic leukaemia, acute lymphoblastic leukaemia, chronic granulocytic leukaemia, chronic lymphocytic leukaemia, Hodgkin's disease and non‐Hodgkin's lymphoma. There was no correlation between the raised GSH levels and decreased Pyr 5′‐NT levels.
Prenatal Diagnosis of β Thalassaemia by Fetal Red Cell Enrichment with NH4Cl‐NH4HCO3 Differential Lysis of Maternal Cells
Authors: Furbetta M. , Angius A. , Ximenes A. , Tuveri T. , Rosatelli C. , Scalas M. T. , Fais R. , Cao A. , And G. Angioni , Caminiti F.
Published: March 2008

Summary. Prenatal diagnosis with globin chain synthesis analysis on fetal red blood cells concentrated by NH4Cl‐NH2HCO3 differential lysis of maternal cells (Ørskov lysis) was carried out in 27 pregnancies at risk for β thalassaemia and one at risk for sickle cell β0 thalassaemia. The β/γ globin chain synthesis ratio was also determined after anti‐i differential agglutination (12 cases), in almost pure fetal samples (six cases) and by extrapolation (one case). Differential lysis permitted the study of samples drawn by placental aspiration containing as little as 3.2% fetal red blood cells. There was no consistent difference between the β/γ ratios observed after differential lysis and those determined after the use of the other approaches. A presumptive diagnosis of homozygous β thalassaemia was made in nine cases. All but one of these pregnancies was terminated. The absence of β chain synthesis was confirmed by the study of fetal blood after abortion in four cases with suitable samples. Of the remaining pregnancies, six proceeded to term and non‐homozygous infants were delivered. The others are still in progress. No fetal loss occurred. ørskov lysis seems to be a very reliable method for prenatal diagnosis of β chain abnormalities. Moreover it can minimize the number and duration of placental aspirations required and thus the risk to the fetus.
The Anaemia of P. falciparum Malaria
Authors: Abdalla S. , Weatherall D.J. , Wickramasinghe S. N. , Hughes Madeleine
Published: March 2008

Summary.

The haematological changes in a group of young Gambian children with P. falciparum malaria have been analysed. In children with acute infection anaemia was most marked during the period after treatment. Although many of these patients developed a positive direct Coombs test during this period of the illness it is not clear whether the anaemia which occurs after treatment has an immune basis. A second group of children showed quite different haematological findings. They appear to have a more chronic form of P. falciparum malaria infection, were profoundly anaemic at presentation, showed gross dyserythropoietic changes in their bone marrows, and had a full reticulocyte response and rise in haemoglobin after treatment. A third group of children were encountered whose haematological abnormalities were intermediate to those of the acute and chronic groups. These findings indicate that the patho‐physiological mechanisms responsible for the anaemia ofP. falciparum malaria are different at different stages of the illness.
Iron Binding Proteins in Selected Human Peripheral Blood Cell Sets: Immunofluorescence
Authors: Nishiya Koji , Chiao Jen Wei , Sousa Maria De
Published: March 2008

Summary.

Selected populations of human peripheral blood T lymphocytes, B lymphocytes, monocytes and polymorphonuclear cells were examined for their intracellular content of lactoferrin transferrin and ferritin by an indirect inimunofluorescence technique. Lactoferrin was found in polymorphs but not in lymphoid cells. Two different lactoferrin staining patterns were observed which we designated ‘perinuclear’, characterized by a ring of positive material round the nucleus, and cytoplasmic, in which most positive material was distributed in the cytoplasm. The former staining pattern was found in high density polymorphs the latter was associated with low density polymorphs occasionally found contaminating the peripheral blood mononuclear cell suspension. After incubation in vitro, the peri‐nuclear pattern changed to cytoplasmic staining.

Transferrin was found in T cells and polyniorphs. In contrast, only a few transferrin containing cells were detected in the B cell and monocyte fractions. Following overnight incubation, a halo of positive material was found surrounding T cells stained for transferrin, suggesting that T cells released transferrin during incubation. Ferritin was also found in T cells and adherent cells. Following latex particle ingestion, the intensity of ferritin staining was markedly increased. Only a small proportion of lymphoid cells and monocytes stained for transferrin or ferritin in the total peripheral blood mononuclear cell suspension. The results indicate, therefore, that in response to manipulation procedures used routinely for the selection of human peripheral blood lymphoid cells, detectable amounts of transferrin and ferritin are present in T but not B cells. The fact that T cells are equipped with proteins known to participate in binding and storage of iron may constitute, at least in part, the basis for the contribution of these cells to the regulation of other major biological systems.
The Genetic Basis of Hb Q‐H Disease
Authors: Higgs D. R. , Hunt D. M. , Drysdale H. C. , Clegg J. B. , Pressley L. , Weatherall D. J.
Published: March 2008

Summary.

A Chinese family has been studied in which two siblings have haemoglobin Q‐H disease. Using a combination of haematological and haemoglobin analysis, globin chain synthesis, analysis of α/β globin messenger RNA ratios and restriction endonuclease mapping, it has been shown that each of these siblings has received one chromosome on which both α chain genes have been deleted and another on which there is only a single α chain locus which carries the αQ mutation. Their genotype is thus ——/—αQ. Despite the fact that the haemoglobin Q mutation in this family is carried on a chromosome with a single α chain locus, heterozygous carriers for the variant have only 25% or less haemoglobin Q. Our observations indicate that the molecular basis for haemoglobin Q‐α thalassaemia is similar to that for the common form of haemoglobin H disease in Orientals. Furthermore, they provide clear evidence that the level of an α chain variant in heterozygous carriers is not a reliable reflection of the number of α globin genes.
Another Example of Haemopoietic Chimaerism in Dizygotic Twins
Authors: Bird G. W. G. , Gibson M. , Wingham June , Mackintosh Pauline , Watkins Winifred , Greenwell Pamela
Published: March 2008

Summary.

Another example of haemopoietic chimaerism in dizygotic twins is described. Each twin had two distinct blood cell populations. The red cell populations differed in three blood group systems and in other genetic characters, and the white cell population in HLA types and in XX/XY karyotypes. The biological significance of these findings is discussed with special reference to the relative proportions of the two red and white cell populations, the ABH blood group gene‐specified glycosyltransferase levels, and the HLA types.
Deep Vein Thrombosis and Changes in Coagulation and Fibrinolysis after Gynaecological Operations in Chinese: the Effect of Oral Contraceptives and Malignant Disease
Authors: Tso S. C. , Wong V. , Chan Vivian , Chan T. K. , Ma H. K. , Todd D.
Published: March 2008

Summary.

Of 154 Chinese patients who underwent gynaecological operations, four showed a positive 125I fibrinogen leg scan for venous thrombosis, an overall incidence of 2.6%. In those who were on oral contraceptives and had major pelvic surgery for benign conditions, the incidence was 10.5%; in those who had Wertheim hysterectomy for carcinoma of cervix, it was 6.7%. This confirms the rarity of post‐operative thromboembolism in the Chinese. Fragment E showed a biphasic rise after major operation due to tissue injury and venous thrombosis. In patients with malignancy, the postoperative ‘fibrinolytic shutdown’, represented by decreased plasminogen activator together with increased α1, antitrypsin and C inhibitor levels, was more marked. In addition, α2 macroglobulin level was lower and fell significantly after operation. In patients on oral contraceptives, fragment E levels were higher after surgery and there was no decrease in plasminogen activator levels. Antithrombin III levels did not fall except in three of the four patients with venous thrombosis. A marked increase in fragment E level and a decrease in antithrombin III level might be useful diagnostic markers for postoperative venous thrombosis.
Ultrastructural Cytochemistry of Leukaemic Cells: Characterization of the Early Small Granules of Monoblasts
Authors: O'Brien Maureen , Catovsky D. , Costello Christine
Published: March 2008

Summary An ultrastructural study of blast cells showing either monocytic or granulocytic differentiation was carried out with the acid phosphatase (AP) and myeloperoxidase (MPO) reactions. Eight cases of acute myeloid leukaemia (AML) and three of chronic granulocytic leukaemia in blast crisis were studied. A hitherto unrecognized small lysosomal granule characterized by AP activity and lack of MPO was present in the majority of cells of all six monoblastic leukaemias. These granules ranged from 0·05 to 0·2 μm in size and were distributed throughout the cytoplasm, frequently at the periphery of the cells. A small proportion of monoblasts showed AP reactivity in the Golgi cisternae. Both AP and MPO were positive in the granules of promonocytes; however, MPO positive granules were predominant in late promonocytes.

Larger granules (0·2–0·6 μm) with MPO reactivity were characteristic of myeloblasts. In only two out of four cases did these granules show AP positivity, suggesting that, in contrast to monoblasts, AP activity is a late feature of myeloblastic differentiation. This study shows that ultrastructural cytochemistry may be helpful in the recognition and classification of acute leukaemias by demonstrating the early differentiation features of monocytic and granulocytic precursors.
The Increased Susceptibility of Young Red Cells to Invasion by the Malarial ParasitePlasmodium falciparum
Authors: Pasvol G. , Weatherall D. J. , Wilson R. J. M.
Published: March 2008

SummaryThe relationship between red cell age and susceptibility to invasion by the malarial parasitePlasmodium falciparumhas been examined by several different methods including short‐term cultures of parasitized human blood. The results indicate that reticulocytes and young red cells are more susceptible to invasion by this parasite as compared with metabolically older cell populations. This is contrary to the current belief that red cells of all ages are invaded indiscriminately byP. falciparum.

This observation has important theoretical, clinical and practical implications; its mechanism remains as yet unclear.
Autoimmune Thrombocytopenia: Detection of Platelet Autoantibodies with the Suspension Immunofluorescence Test
Authors: von dem Borne A. E. G. Kr. , Helmerhorst F. M. , van Leeuwen E. F. , Pegels H. G. , von Riesz E. , Engelfriet C. P.
Published: March 2008

Summary An immunofluorescence test on paraformaldehyde‐fixed platelets in suspension was used for the detection of antibodies on platelets and in the sera of patients with idiopathic or secondary thrombocytopenia.

In idiopathic thrombocytopenia, a positive test was found on the patients' platelets in 55 out of 80 cases. The indirect immunofluorescence test with the patient's serum was positive in 28 out of 80 cases. In thrombocytopenia accompanied by other autoimmune diseases, the direct test was positive in 35 out of 39; the indirect test in 13 out of 39 patients. These results are similar to those obtained with other techniques recently described in the literature.

By using FITC‐labelled monospecific anti‐immunoglobulin reagents, immunochemical characterization of the autoantibodies was possible. Of the 90 patients with a positive direct test, 80 were analysed in this way. Of these, platelet‐bound IgG was detected in 76. This was accompanied by IgM in 17 and IgA in three. In four, only IgM was found. The IgG subclasses of platelet‐bound IgG were studied in 48 patients. Mainly IgG1 was found (45), often together with IgG3 (22) or IgG4 (14), but rarely with IgG2 (1). Only a few patients had just IgG3 (2) or IgG4 (1) on their platelets.

Platelet‐bound autoantibodies could often be eluted (41 out of 86 cases) with ether and were demonstrable in the eluate with the indirect immunofluorescence test. This test was usually positive when the direct test was strongly positive (40 out of 44) and usually negative when it was weakly positive (1 out of 24) or negative (1 out of 18).
Variation in Sensitivity of DNA Synthesis to Ara‐C in Acute Myeloid Leukaemia
Authors: Harris Adrian L. , Grahame‐Smith D. G.
Published: March 2008

Summary The sensitivity of DNA synthesis to ara‐C was measured in vitro in intact myeloblasts from patients with acute myeloid leukaemia (AML). Ara‐C concentrations of 1 nm to 100 μm were used and ara‐CTP production measured at the same concentrations. Maximum inhibition of DNA synthesis was 95% in all patients. At ara‐C concentrations above 1 μm there was little variation in sensitivity of DNA synthesis but at concentrations below 300 μm there was wide variation in inhibition. Ara‐CTP concentrations associated with 50% inhibition of DNA synthesis ranged from 0·034 to 1·0 pmol/106 cells. Variation in sensitivity of DNA synthesis to ara‐C could not be explained by differences in ara‐CTP production alone. Concentrations of 100 nm to 1 μm ara‐C in vivo may be desirable to produce greater than 50% inhibition of DNA synthesis in all patients’myeloblasts.
Lymphoid Cell Sets and Serum Immunoglobulins in Patients with Thalassaemia Intermedia: Relationship to Serum Iron and Splenectomy
Authors: Kapadia Asha , de Sousa Maria , Markenson Alicejane L. , Miller Denis R. , Good Robert A. , Gupta Sudhir
Published: March 2008

Summary Peripheral blood lymphocyte sets and serum immunoglobulins were examined in 15 patients with thalassaemia intermedia, aged between 5 and 30 years. They were divided in three clinical groups: Group I, non‐splenectomized, with serum iron (SI) lower than 200 μg/dl; Group II, splenectomized with serum iron lower than 200 μg/dl; and Group III, splenectomized with serum iron levels greater than 200 μg/dl. High absolute lymphocyte counts were observed in the majority of patients, whether splenectomized or not. Following splenectomy, marked increases were observed in absolute number of lymphocytes and percentages of circulating surface immunoglobulin (SIg) bearing B lymphocytes, mouse rosette forming cells (MRFC) and ‘null’ cells. The increases in circulating B lymphocytes following splenectomy were due exclusively to cells bearing SIgM and/or SIgD. No changes were observed in numbers and proportions of circulating SIgA or SIgG bearing cells. Serum IgA levels were high in the majority of patients studied whether splenectomized or not. Following splenectomy increases in serum IgA and serum IgG were observed which seemed to relate to increasing SI levels. No significant changes occurred in serum IgM levels.

Absolute numbers of circulating T lymphocytes were unaffected by splenectomy or SI levels. Changes were observed, however, in distribution of T lymphocyte subsets with decreases in number of Tμ cells and concomitant increases in numbers of non‐Tμ, non‐Tγ cells. Tγ cells were the least affected by splenectomy and increasing SI. The results are compatible with the proposition that iron participates in the regulation of lymphoid cell migration and in the modulation of expression of cell surface markers. The findings also illustrate the role of the spleen in the control of lymphocyte migration and immunoglobulin production.
Hb F Synthesis in Sickle Cell Anaemia: a Comparison of Saudi Arab Cases with those of African Origin
Authors: Wood W. G. , Pembrey M. E. , Serjeant G. R. , Perrine R. P. , Weatherall D. J.
Published: March 2008

Summary Fetal haemoglobin (Hb F) synthesis has been studied in 22 cases of sickle cell anaemia (SS) from Saudi Arabia and compared with an equal number of cases of African origin. Among the Saudi Arabs γ chain synthesis ranged from 4·0% to 19·9% of the total non‐α chain synthesis (mean 8·1%) while the corresponding range for the Negro cases was <0·3% to 4·6% (mean 1·7%). In both groups the peripheral blood Hb F level was on average 3–4 times higher than the proportion synthesized, indicating that the selective survival of Hb F containing cells (F cells) was an important factor in determining the final Hb F levels. Among the Saudi Arab cases there was a significant negative correlation between the degree of F cell enrichment and either the Hb F level or the percentage γ chain synthesis. No such correlation was observed among the Negro cases. A high proportion of the cases in both groups were carriers of α thalassaemia in addition to SS, but no effect of α thalassaemia on Hb F production was observed.
Histological Study of Bone Marrow Regeneration following Chemotherapy for Acute Myeloid Leukaemia and Chronic Granulocytic Leukaemia in Blast Transformation
Authors: Islam Anwarul , Catovsky Daniel , Galton David A. G.
Published: March 2008

Summary Haemopoietic regeneration following chemotherapy was studied on serial bone‐marrow trephine specimens from 10 patients with acute myeloid leukaemia (AML), and seven with chronic granulocytic leukaemia (CGL) in blast transformation. In AML following the stage of treatment‐induced hypoplasia during which the marrow was extremely hypocellular, oedematous, and contained widely dilated sinuses, areas of large, uniform, unilocular fat cells, designated ‘structured fat’ developed from multilocular precursor fat cells. Early foci of haemopoietic regeneration were present almost exclusively in areas of structured fat: only scattered small erythropoietic foci were seen in the earliest specimens taken 1 week after the completion of treatment; small granulopoietic foci were first seen 2 weeks later, and the first megakaryocytes were not seen until the third week. Transient regeneration was observed in two patients who subsequently failed to enter remission and died.

The pattern of haemopoietic regeneration in CGL in blast transformation treated by intensive therapy and autografting with cryopreserved buffy‐coat cells differed from that seen in AML. Bone marrow regeneration occurred earlier than in AML and appeared to be independent of the presence of structured fat. Regeneration in all three cell lines appeared simultaneously within 2 weeks of autografting. Regeneration tended to be focal, each aggregate consisting almost exclusively of erythroid, granulocytic or megakaryocytic cells.

The proximity of regenerating haemopoietic foci to structured fat in AML suggests that, in some circumstances, haemopoietic stem cells require the presence of fat cells in their immediate environment before they proliferate.