bluuebunny/crossref_metadata_2025 huggingface dataset:
Interferon Production in Leukaemia
Authors: Chisholm Morag , Cartwrigih T.
Published: March 2008

SUMMARY Leucocyte interferon production in vitro and circulating interferon levels were studied in healthy subjects and in 80 patients with acute or chronic leukaemia. Circulating interferon was not found in either group. Interferon synthesis in response to a virus was normal in patients with acute leukaemia and appeared to be enhanced in some. In chronic leukaemia reduced levels were common particularly in CLL, in which condition normal results were rarely found; lymphocyte transformation to PHA was also depressd in this group. No clinical or haematological correlation with the interferon levels was found and no consistent effect of treatment was shown. The possible factors which could account for these findings arid their significance in relation to pathogenesis and treatment of leukaemia are discussed.
Erythrokinetic Studies in Myelofibrosis: Their Significance for Prognosis
Authors: Najean Yves , Cacchione Roberto , Castro‐Malaspina Hugo , Dresch Catherine
Published: March 2008

Summary. Eighty‐three patients with myelofibrosis have been studied by erythrokinetics and have been followed up until death or for at least 12 months. Because of a large plasma volume the venous haematocrit gives only a poor idea of the red blood cell volume. The red cell survival was reduced in the majority of cases but significant haemolysis was rare. The amount of haemolysis of autologous and isologous red cells was similar, suggesting an extra‐corpuscular origin for the haemolysis. Plasma iron turnover was always increased, sometimes markedly, but red cell iron incorporation was reduced in 70% of cases, indicating ineffective erythropoiesis. Surface counting showed an absence on diminution of sacral iron fixation and a rapid and marked splenic uptake in more than 90% of the cases; uptake of iron by the liver occurred in half the cases, usually not very high; iron release from the spleen was absent or reduced in 67% of the cases.

The degree of ineffective erythropoiesis as measured by radio‐iron incorporation and release by the spleen, the amount of haemolysis, and the red cell volume were strongly correlated with prognosis. These factors enabled a more precise prediction to be made of the clinical outcome in the 2 years following the study, than the clincal data alone. A prospective study might show whether erythrokinetic studies are also useful in determining the choice of treatment.
Constitutional Aplastic Anaemia: Defective Haematopoietic Stem Cell Growth in Vitro
Authors: Saunders E. F. , Freedman M. H.
Published: March 2008

Summary. . Seven children with constitutional aplastic anaemia (one dyskeratosis congenita, one constitutional type without anomalies, five classical Fanconi's anaemia) were studied to define the pathogenesis of the marrow failure. Haematopoiesis was assessed by assay of granulocytic (CFU‐C) and erythrocytic (CFU‐E) colonies in vitro.

Numbers of CFU‐C were profoundly reduced or absent in marrow cultures from all patients (range 0‐5 CFU‐C/105, control range 15‐106/105). Numbers of CFU‐E were also strikingly reduced or absent (range 0‐2 CFU‐E/105, control range 31‐317/105) with one exception. No CFU‐C could be grown from peripheral blood. Incubation of the patients' sera with control marrow failed to demonstrate inhibition of CFU‐C, colony stimulating activity (CSA), CFU‐E or erythropoietin (EPO). The peripheral blood lymphocytes from these patients in co‐culture experiments failed to inhibit CFU‐C or CFU‐E growth from control marrow. Aplastic marrow co‐cultured with control marrow also failed to demonstrate cellular inhibition. CSA production from patients’peripheral blood leucocytes averaged 93% of the activity of a standard control CSA, suggesting normal granulopoietic stimulation.

The data indicate a problem early in haematopoiesis. Although a defect in the stromal microenvironment cannot be excluded, it is possible the defect is intrinsic to the haematopoietic stem cells, either an absence or marked reduction in stem cells, or a failure of stem cells to grow colonies in Vitro.
Use of a Simple Visual Assay of Willebrand Factor for Diagnosis and Carrier Identification
Authors: Reisner H. M. , Katz H. J. , Goldin L. R. , Barrow E. S. , Graham J. B.
Published: March 2008

Summary. A visual assay of factor VIII‐related Willebrand factor (VIIIR:WF) is described which utilizes formaldehyde‐fixed platelets, end points being read in microflocculation tiles. Four dilutions of a sample can be assessed simultaneously, and the correlation with aggregometric assays is high (r=0.91). Measurement error is 8.0% for a single assay in triplicate and less than 5% if an assay is repeated three times. The method has been used for 2 years by the coagulation genetics group at Chapel Hill for diagnosing subjects with von Willebrand's disease and assigning genotypes to members of families transmitting this disorder. Its utility in classifying known carriers of haemophilia A has also been examined, both in conjunction with assays of VIII:C and in a three‐way test with assays of VIII:C and VIIIR:Ag. As predicted by the Lyon hypothesis, the rate of false negative diagnosis was higher than false positive diagnosis, but the overall rate of misclassification on single plasma samples was 7/51=13.7%. The error rate was the same whether discrimination was based upon assays of VIII:C vs. VIIIR:Ag, VIII:C vs. VIIIR:WF, or VIII:C vs. VIIIR:Ag vs. VIIIR:WF, the same individuals being misclassified by each method. The observed rate of misclassification was well within the rates reported by others and very similar to our previous experience. We have concluded that this method of assaying VIIIR:WF is highly useful for diagnosing vWd, detecting inhibitors to VIIIR:WF, and examining large numbers of column fractions. It is a useful supplement, although it cannot yet substitute for, assays of VIIIR:Ag in detecting carriers of haemophilia A.
Changes of Granulopoiesis during and after Adjuvant Chemotherapy of Breast Cancer
Authors: Lohrmann Hans‐Peter , Schreml Wolfgang , Lang Margarete , Betzler Michael , Fliedner Theodor M. , Heimpel Hermann
Published: March 2008

Summary. Adjuvant chemotherapy allows a study of the effects of cytotoxic drugs on natural human haematopoiesis. We describe serial studies of granulopoiesis performed during and after intermittent adjuvant chemotherapy for breast cancer (adriamycin plus cyclophosphamide, given for six courses at monthly intervals). After drug administration, a sequential wave of depletion and regeneration through successive granulopoietic compartments was observed. With repeated chemotherapy, moderate neutropenia developed, and the blood CFU‐C pool size became progressively reduced. After the sixth chemotherapeutic course, granulopoietic regeneration was delayed. Following discontinuation of chemotherapy, a long‐lasting (>200 d) reduction of the blood CFU‐C pool size, together with neutropenia and reduction of marrow segmented neutrophils, was observed, suggesting a defect of granulopoiesis with persistent granulopoietic hypoplasia. In patients with expected long survival, residual bone marrow damage should be added to the list of potential late side effects of cytotoxic drug therapy.
Formation of Transcobalamin II‐Vitamin B12 Complex by Guinea‐Pig Ileal Mucosa in Organ Culture After in Vivo Incubation with Intrinsic Factor‐ Vitamin B12
Authors: Rothenberg Sheldon P. , Weiss James P. , Cotter Richard
Published: March 2008

Summary. The in vivo incubation of intrinsic factor‐[57Co]vitamin B12 in an ileal loop of a guinea‐pig followed of in vitro culturing of segments of the ileum for 180 min has been used to study the transepithelial transport vitamin B12. Analysis of the solubilized supernate of mucosa following the in vivo phase demonstrated that 44% of the [57Co]vitamin B12 was bound to intrinsic factor (IF), 26% was free, and 16% was bound to transcobalamin II (TCII). Following culture, similar analysis demonstrated that 18% of the vitamin was now bound to IF, 49% was free, and 35% was bound to TCII. In the culture medium, 54% of the [57Co]vitamin B12 was free and 37% was bound ot TCII. The formation of TCII‐[57]vitamin B12 did not occur if homogenized mucosa was incubated with free [57Co]vitamin B12 but it did form in cultures of ileal segments from animals given an excess of unlabelled vitamin to saturate all circulating TCII, and in the medium containing puromycin. Indirect immunoflourescence using chicken anti‐TCII demonstrated that TCII was associated with the mucosal cells of both the ileum and jejunum.

These studies demonstrate that following transepithelial flux of vitamin B12 through the ileal mucosa, the vitamin becomes coupled to TCII. This coupling requires a structurally intact mucosa and the source of the TCII appears to be the ileal mucosal cell rather than unsaturated TCII circulating in the blood.
Energy Reserve and Cation Composition of Irreversibly Sickled Cell in Vivo
Authors: Glader B. E. , Lux S. E. , Muller‐Soyano A. , Platt O. S. , Propper R. D. , Nathan D. G.
Published: March 2008

Summary. Cation composition, cellular hydration, and adenosinetriphosphate (ATP) content were measured in irreversible sickle cells (ISC's) separated from the blood of patients with sickle cell anaemia. Total monovalent cation (Na++K+) content was markedly reduced in ISC's and this largely was due to cell K+ depletion. Corresponding to the reduced cation content, cells were dehydrated as indicated by a reduced mean cell volume. ISC's also appeared to be grossly depleted of ATP. These biochemical characteristics allow us to expand the definition of ISC's beyond morphologic characteristics. In addition, these chemical alterations provide a means for elucidating the mechanism of ISC production in vitro.
Characterization of Serum Fibrinogen and Fibrin Fragments Produced during Disseminated Intravascular Coagulation
Authors: Lane D. A. , Preston F. E. , Van Ross M. E. , Kakkar V. V.
Published: March 2008

Summary. The fibrinogen and fibrin degradation products (FDP) in serum samples taken from nine patients with suspected disseminated intravascular coagulation have been characterized using a method of immunoprecipitation followed by sodium dodecyl sulphate polyacrylamide gel electrophoresis. All of the serum samples contained a fragment with the same electrophoretic mobility as fibrinogen fragment X, while the majority also had evidence of fragments with similar mobility to fibrinogen fragments Y and D. In eight of the nine serum samples there was strong evidence of the D‐dimer fragment that is released by plasmin lysis of crosslinked fibrin. Also present in all but one of the samples were fragments of higher molecular weight than fibrinogen which were probably soluble, non‐clottable, factor XIIIa induced crosslinked derivatives of fibrinogen. These results suggest that during disseminated intravascular coagulation thrombin and activated factor XIII act upon fibrin(ogen) to form complexes that are subsequently lysed by plasmin to produce soluble crosslinked derivatives of fibrin.
Immunological Heterogeneity of Haemophilia B: a Multicentre Study of 98 Kindreds
Authors: Parekh V. R. , Mannucci P. M. , Ruggeri Z. M.
Published: March 2008

Summary. An electroimmunoassay with a precipitating rabbit anti‐human factor IX antiserum and an inhibitor neutralization assay with a non‐precipitating homologous antibody were used to measure factor IX antigen (IX:Ag) in 117 patients from 98 kindreds with haemophilia B; and to investigate in a mixed population the incidence of different immunological types of the disease. Although the two assays showed an excellent correlation, the electroimmunoassay was selected for its simplicity as a criterion for classification. 52 kindreds, referred to as haemophilia B‐, were characterized by severe deficiency of factor IX coagulant activity (<0.01‐0.03 u/ml) and unmeasurable IX:Ag (< 0.12 u/ml): this genetic variant of the disease appears to be related to a complete or marked suppression of factor IX synthesis. In 16 kindreds, a severe or moderately severe IX:C deficiency was associated with normal or increased levels of IX:Ag (haemophilia B+): among them, a subgroup of five kindreds could be identified by the additional abnormality of a prolonged Thrombotest clotting time (haemophilia BM). These patients are likely to be the expression of normal or increased synthesis of a factor IX molecule markedly defective in the site(s) responsible for coagulant activity. Reduced levels of IX:Ag (0.12‐0.65 u/ml) characterized the remaining 30 kindreds, presenting with IX: C levels ranging from < 0.01 to 0.21 u/ml. In 28 there was a significant excess of IX:Ag over IX:C, suggesting a reduced capacity to synthesize the factor IX molecule accompanied by a variable defect in the coagulant site; the remaining two kindreds, which showed a concomitant reduction of IX:C and IX:Ag, are likely to be examples of a true reduction of factor IX synthesis.
Critical Re‐examination of the Specificity of Auto‐anti‐Rh Antibodies in Patients with a Positive Direct Antiglobulin Test
Authors: Issitt Peter D. , Pavone Beverly G.
Published: March 2008

Summary

Forty‐eight autoantibodies with apparent 'simple’anti‐Rh specificity (anti‐e, ‐E, ‐c, ‐D, ‐C, ‐Ce, ‐G), have been studied by means of multiple absorption tests. The finding that 34 (70.8%) of these antibodies could bind to red blood cells lacking the antigens that the antibodies appeared to define, indicated that the antibodies had different specificities than seemed to be the case in initial antibody identification tests. Those autoantibodies that at first appeared to be directed against the Rh antigens e, E or c, most often had anti‐Hr or anti‐Hro specificity.

These data explain why some apparent anti‐Rh autoantibodies can be eluted from the red blood cells of patients negative for the antigens that the antibodies appear to define. However, they also illustrate that the phenomenon of autoantibodies mimicking specificities that they do not possess is common in patients positive for the antigens against which their autoantibodies appear to be directed. An explanation for the mode of action of these autoantibodies in complexing with the Rh agglutinogen is proposed, and the significance of the antibodies in transfusion therapy is considered.
Characteristic Surface Morphology of Human and Murine Myeloma Cells: a Scanning and Transmission Electron Microscopic Study
Authors: Polliack Aaron , Nilsson Kenneth , Laskov Reuven , Biberfeld Peter
Published: March 2008

Summary. Cells from cultured human and murine myeloma cell lines and circulating leukacmic plasma cells from four patients with generalized myeloma were studied by transmission and scanning electron microscopy and time‐lapse cinematography. Both circulating and cultured cells exhibited consistent surface architectures and varying numbers of prominent blebs of different sizes were seen, in addition to microvilli. The presence of surface blebs appears to be a characteristic feature for secreting and non‐secreting myeloma cells.
The Inhibition of Plasmin by Antithrombin–Heparin Complex
Authors: Semeraro N. , Colucci M. , Telesforo P. , Collen D.
Published: March 2008

Summary. Human plasma, containing increasing amounts of heparin (0‐125 iu/ml) and highly purified biologically intact131 I‐labelled antithrombin was activated with 2000 CTA units of urokinase per ml, which resulted in complete activation of the fibrinolytic system. The plasmin‐antithrombin‐(heparin) complex formed was isolated and quantitated. In the absence of heparin only 0.4 ± 0.1% of the plasma antithrombin was bound to plasmin. In the presence of heparin it was: 0.9±0.1% with 1 iu/ml, 1.5±0.7% with 2 iu/ml, 2.5±0.8% with 5 iu/ml, 3.8 ± 0.5% with 25 iu/ml and 4.7±1.0% with 125 iu/ml. Thus antithrombin–heparin complex seems to play a very limited role in the inactivation of plasmin under physiological conditions and therapeutic levels of heparin. In plasma depleted of antiplasmin, the fast‐reacting plasmin inhibitor, 1.6–2.1 % of antithrombin was bound to plasmin in the absence of heparin, and 8.2‐10% in the presence of 25 iu heparin/ml. In plasma depleted both of antiplasmin and α2‐macroglobulin, 8.1–8.6% of antithrombin bound to plasmin in the absence of heparin and 16–21 % in the presence of 25 iu of heparin/ml. However, since the molar concentration of plasminogen in plasma is approximately half that of antithrombin, the relative amount of plasmin neutralized by antithrombin is twice as high.
Microheterogeneity of Human Antithrombin III
Authors: Borsodi A. , Narasimhan T. R.
Published: March 2008

Summary. Antithrombin purified from normal human plasma has been separated into two fractions by isoelectric focusing in a pH 4‐6 gradient. These fractions were homogeneous by polyacrylamide gel electrophoresis, had similar amino acid composition and the same specific activity. Both of them cross reacted with antiserum against antithrombin. They were found to contain different amounts of sialic acid and aminosugars. After neuraminidase treatment only a single, homogeneous peak was found by isoelectric focusing–with unchanged antithrombin activity–suggesting that the microheterogeneity is due to a difference in glycosylation.
An Immunoradiometric Assay for Procoagulant Factor VIII Antigen: Results in Haemophilia, von Willebrand's Disease and Fetal Plasma and Serum
Authors: Peake I. R. , Bloom A. L. , Giddings J. C. , Ludlam C. A.
Published: March 2008

Summary. An immunoradiometric assay (IRMA) has been developed based on the inhibitor which arose in a polytransfused severe haemophiliac. The two‐site IRMA measures antigens closely associated with the procoagulant parts of the factor VIII complex, which are termed FVIIIC antigens or FVIIICAG. FVIIICAG was present in normal plasma and also, at a slightly lower concentration, in normal serum. In 37 patients with haemophilia A, 36 had FVIIICAG levels of less than 10% of the normal plasma pool. In patients with von Willebrand's disease the levels of FVIIIC and FVIIICAG were in good agreement, both before and after treatment with cryoprecipitate or DDAVP. FVIIICAG was relatively stable in plasma at 37°C and could also be detected in cord and fetal serum. The assay is of potential value for detecting reduced levels of factor VIII, for carrier detection and for the prenatal diagnosis of haemophilia.
The Inhibitory Effect of Aspirin on Platelet and Vascular Prostaglandins in Rats cannot be Completely Dissociated
Authors: Villa Silvia , Livio Manuela , de Gaetano Giovanni
Published: March 2008

Summary. Blood platelets and vascular endothelium generate prostaglandin derivatives with opposing effects: cyclic endoperoxides and thromboxane A2 (TXA2) aggregate platelets, whereas prostaglandin I2 (PGI2, prostacyclin) inhibits aggregation. Since aspirin inhibits the synthesis and effects of all these metabolites of arachidonic acid, the rationale for clinical use of this drug in thrombosis prevention trials is questionable. Treatment schedules would have to be calibrated to inhibit proaggregating platelet prostaglandins and TXA2 without affecting protective vascular prostaglandins (PGI2). This important clinical information cannot easily be obtained from studies in man.

The present study indicates that normal rat platelets are more sensitive to aspirin (ID50=3.6 mg/kg) than arterial tissues (ID50=25.0 mg/kg) and are almost as sensitive as venous tissues (ID50=2.3 mg/kg). Platelet prostaglandin production was monitored through the measurement of malondialdehyde (MDA) output under standard stimulation with thrombin. PGI2‐like activity released from the abdominal aorta and inferior vena cava was determined as platelet aggregation inhibitory potency and characterized by standard criteria.

The inhibitory effect of aspirin (at doses of 10 and 50 mg/kg) on platelet MDA production lasted much longer in platelets (96–120 h) than in venous (24–72 h) or arterial tissue (less than 24 h). The findings suggest that in rats the (potentially antithrombotic) effect of aspirin on platelet prostaglandin and TXA2 generation may be distinguished, though not completely, from the (potentially prothrombotic) effect on vascular PGI2 on the basis of duration of inhibition more than on the sensitivity of the target cells to the drug. The different capacities of anucleated platelets and of nucleated vascular endothelial cells to overcome aspirin inhibition could be exploited for more rational clinical use of this drug.
Detection of Haemophilia A Carriers by Replicate Factor VIII Activity and Factor VIII Antigenicity Determinations
Authors: Seligsohn Uri , Zivelin Ariela , Perez Chava , Modan Michaela
Published: March 2008

Summary. Factor VIII (fVIII) activity and antigenicity were determined on replicate fresh blood samples drawn on three separate occasions in 48 normal females and 37 obligatory haemophilia A carriers, in an attempt to improve the detection rate. fVIII activity was assayed by a one‐stage method and fVIII antigenicity by the Laurell's technique employing an anti‐fVIII antiserum prepared in rabbits. The mean results for the individual subjects were analysed after establishing a discriminant function. The probability of being a carrier was determined on the basis of laboratory data and family history. The probability data were translated into more simple terms applicable for genetic counselling by defining three categories: Definite carriers, doubtful carriers and definite normals. Of the obligatory carriers, 83·8% were classified definite carriers, 13·5% doubtful carriers and 2·7% as definite normals. Of the normal females, 4·2% were classified as definite carriers, 18·7% doubtful carriers and 77·1% as definite normals. Adjustment of the results according to age was found unnecessary.

The precision of the determinations of fVIII antigenicity and fVIII activity in individual subjects was calculated for different numbers of replicate tests performed in the same individual. It was found that if only one test is performed, a substantial error might occur. Three replicate tests considerably diminish this error, although it is still in the range of ±16–29%. For suspected carriers who fall in the doubtful category it is suggested that six or even more tests are performed.
Coagulation Failure in Babies with Rhesus Isoimmunization
Authors: Hey E. , Jones P.
Published: March 2008

Summary. Serial prospective studies of coagulation status have been undertaken on 73 babies with a positive Coombs test. No abnormalities were detected in the babies with mild haemolytic disease, but seven of the 36 babies with severe haemolytic disease (cord Hb < 11 g/dl or cord bilirubin > 85 μmol/l) showed evidence of transient defibrination 1 d after birth and another six had evidence of coagulation failure at birth with a platelet count of less than 150 × 109/l and a severe deficiency of multiple coagulation factors. The level of factor II and factor X was less than a fifth of the normal cord blood level in these six babies and the level of I, VII and IX was severely reduced; the factor VIII level was normal or high. Exchange transfusion started within 1 h of birth corrected the immediate factor deficiency in these six babies, but evidence of defibrination then became apparent with afibrinogenaemia, a marked fall in factors II and V, less constant falls in factors VII, IX and X, and a raised fibrin: fibrinogen degradation product level. One of these six babies died with severe pulmonary hypoplasia within an hour of birth; the other five died from haemorrhage into the lung or brain 1 1/2–6 d after birth. The very low vitamin‐K dependent factor levels in the cord blood of the babies who died are presumably the result of liver damage in utero, but the subsequent changes are those of a comsumption coagulopathy. Simple screening tests at birth served to indicate which babies were at risk and it is concluded that death due to haemorrhage might be reduced by more intensive factor replacement before there is overt evidence of haemorrhage in these babies.
The Separation of Willebrand Factor from Factor VIII‐Related Antigen
Authors: Barrow E. S. , Reisner H. M. , Graham J. B.
Published: March 2008

Summary. The three activities associated with factor VIII—coagulant (VIII:C), antigenic (VIIIR:Ag), and platelet agglutinating or Willebrand factor (VIIIR:WF)— have been separated by sequential antibody affinity chromatography, utilizing a rabbit antibody to factor VIII and a spontaneous human antibody to VIII:C. Normal plasma differentially lost its factor VIII‐related antigen following passage over the rabbit antibody column. Subsequent passage of the VIIIR:Ag‐depleted plasma over the human antibody column resulted in the loss of VIII:C activity, with retention of the Willebrand factor activity, antigen being partially recovered from the heterologous antibody column. These experiments demonstrate that it is possible to separate two of the factor VIII activities, VIIIR:Ag and VIIIR:WF, which are usually regarded as properties of a single molecule.
Association of IgA Multiple Myeloma with Pre‐existing Disease
Authors: Schafer A. I. , Miller J. B.
Published: March 2008

Summary. A retrospective analysis of 153 patients with multiple myeloma was performed for evaluation of the possible significance of pre‐existing disease. 37% of the group had no significant antecedent disorder. The most common prior illnesses were peptic ulcer disease and gallbladder disease. Of 12 patients in the group who had prior biliary tract disease and for whom immunoelectrophoretic studies were available, eight (66.7%) had IgA paraproteins. This figure is statistically higher (P<0.01) than the 14.1% prevalence of IgA paraproteins in those myeloma patients without biliary disease. Of 20 individuals with IgA myeloma, only two had no significant antecedent disease. The rest had primarily chronic biliary, peptic ulcer, other gastrointestinal or respiratory tract inflammatory diseases. We conclude that prior inflammatory gastrointestinal, pulmonary, and, particularly, biliary disease may be implicated in the pathogenesis of the IgA subset of multiple myeloma.
Defective Hydrogen Peroxide Production in Chronic Granulocytic Leukaemia Neutrophils
Authors: El‐Maallem Hilal , Fletcher John
Published: March 2008

Summary. Hydrogen peroxide (H2O2) production by polymorphonuclear neutrophils (PMN) during phagocytosis was fluorometrically estimated over an incubation period of 80 min during which dead Candida guilliermondii were ingested. Ten patients with chronic granulocytic leukaemia (CGL) were tested and compared with 15 normal subjects. The production of H2O2 by CGL neutrophils was about 29% of normal. There was no correlation between H2O2 production and the impaired killing of Candida guilliermondii by the neutrophils of CGL patients.
Comparison of Sickle Cell‐β° Thalassaemia with Homozygous Sickle Cell Disease
Authors: Serjeant G. R. , Sommereux AnneMarie , Stevenson Marianne , Mason Karlene , Serjeant Beryl E.
Published: March 2008

Summary. Clinical and haematological features in 41 patients with sickle cell‐β° thalassaemia (Sβ° thalassaemia) and in 123 age‐sex matched controls with homozygous sickle cell (SS) disease were compared. Persistence of splenomegaly was more common and fetal loss less common in Sβ° thalassaemia but other clinical features were similar in the two genotypes. Total haemoglobin, Hb A2, PCV, CCV, and red cell count were significantly higher and MCV, MCH, MCHC, and ISC counts significantly lower in Sβ° thalassaemia. Proportional reticulocyte counts were significantly lower in Sβ° thalassaemia but there was no difference in absolute reticulocyte counts. Persistence of splenomegaly and low ISC counts are compatible with decreased intravascular sickling which may result from the lower mean cell haemoglobin S concentration in Sβ° thalassaemia. If beneficial effects of a low MCHC can be confirmed then a carefully monitored trial of iron deficiency in SS disease may be a logical experimental procedure.
Congenital Non‐spherocytic Haemolytic Anaemia Variants with Primary and Secondary Pyruvate Kinase Deficiency
Authors: Ważewska‐Czyżewska Maria , Gumińska Maria
Published: March 2008

Summary. The patterns of survival of isotope‐labelled erythrocytes were examined in patients suffering from two variants of congenital non‐spherocytic haemolytic anaemia with decreased erythrocyte pyruvate kinase (PK) activity.

In one variant, with primary PK defect (PPKD) randon destruction of erythrocytes was predominant in the process of haemolysis. In the second variant, with primary magnesium activated adenosine triphosphatase (ATP‐ase) (Mg++) deficiency and a secondary decrease in PK activity, erythrocytes were destroyed by senescence. Two subpopulations of labelled erythrocytes with different destruction rates were observed in all patients examined, except one with the second variant, with very mild haemolysis.

Splenectomy, performed on two patients, was successful only in the variant with PPKD.
Congenital Non‐spherocytic Haemolytic Anaemia Variants with Primary and Secondary Pyruvate Kinase Deficiency II. ENZYMATIC STUDIES
Authors: Gumińska Maria , Ważewska‐Czyżewska MARIA
Published: March 2008

Summary. Some metabolic effects associated with defective pyruvate kinase (PK) in two variants of congenital non‐spherocytic haemolytic anaemia with primary PK and primary adenosine triphosphatase (ATP‐ase) (Mg++) deficiency respectively were compared. In one patient with a low erythrocyte ATP level, decreased PK activity appeared together with the irreversible loss of its sensitivity to fructose‐1,6‐diphosphate (FDP), independently of the experimental conditons. In the second patient, the decrease in PK activity associated with an elevated erythrocyte ATP level was a secondary effect, due to primary ATP‐ase (Mg++) deficiency. Removal of excessive amounts of ATP, by dialysis of haemolysates or their in‐vitro treatment with ATP‐ase, increased PK activity to the normal range and restored its sensitivity to the stimulatory effect of FDP. Similar effects could be obtained after i.v. administration of magnesium laevulinate. Under these in vivo conditions the ATP level was normalized after a transient rise in ATP‐ase activity, the PK activity increased and its sensitivity to FDP reappeared.
Altered Erythrocyte Pyrimidine Activity in Vitamin B12 or Folate Deficiency
Authors: Van der Weyden Martin B. , Cooper Mary , Firkin Barry G.
Published: March 2008

The net activity of orotidylic pyrophosphorylase and decarboxylase, sequential enzymes which catalyse the formation of uridine monophosphate from orotic acid in de novo pyrimidine biosynthesis, has been evaluated in erythrocytes of patients with folate or cobalamin deficiency. In patients with normoblastic haemopoiesis and normal cobalamin and folate status a direct relationship exists between the maturity of the peripheral blood erythrocyte population, as indicated by G6PD activity, and net orotidylic activity. In contrast in cobalamin or folate deficiency this co‐ordinate relationship is not observed and net orotidylic activity is relatively reduced. Fractionation of erythrocytes by centrifugation demonstrates that this inordinately low orotidylic activity consistently occurs in the young erythrocyte population and is reversed by specific replacement therapy.
Mechanism of Desferrioxamine‐Induced Iron Excretion in Thalassaemia
Authors: Hershko C. , Rachmilewitz E. A.
Published: March 2008

The mechanism of desferrioxamine (DF)‐induced iron excretion was investigated in 16 thalassaemic patients. Transferrin‐59Fe has been injected intravenously and the fraction of radioiron located to iron stores was determined as the difference between total injected59Fe and % RBC utilization. Following the intravenous infusion of 4 g DF, the specific acitivity of excreted urinary iron has been measured in 11 patients and the total chelatable pool was calculated as (100–% RBC utilization)/specific activity of urinary iron. Despite the wide variation in DF induced urinary iron excretion, ranging from 3 to 86 mg/d, an unexpected uniformity in the specific activity of chelated urinary iron was found in all patients, and the calculated chelatable pool was 4.5±1.2 (mean±SD) g of iron. These findings suggested that iron chelated by DF was probably not obtained directly from the total storage iron pool but rather from a smaller high specific activity iron compartment derived from the catabolism of labelled non‐viable erythrocytes. This postulate was tested by the direct measurement of specific activities in the liver and spleen in five patients undergoing surgery. In all patients, the specific activity of splenic non‐haem iron representing RE iron stores (31±8%59Fe/gFe), was much higher than that of hepatic iron (2±1%), and the specific activity of iron excreted in the urine (30p8%) was identical with the specific activity of splenic iron. These findings indicate that DF‐induced urinary iron excretion is derived from a chelatable pool located in the RE system.
Favism: Erythrocyte Metabolism during Haemolysis and Reticulocytosis
Authors: Gaetani G. F. , Mareni C. , Salvidio E. , Galiano S. , Meloni T. , Arese P.
Published: March 2008

Summary. The reduced activity of glucose‐6‐phosphate dehydrogenase (d‐glucose‐6‐phosphate; NADP+ i‐oxidoreductase; G6PD) in Mediterranean erythrocytes explains the precarious equilibrium of the hexose monophosphate pathway (HMP) and the susceptibility of these cells to haemolytic agents. G6PD‐deficient erythrocytes, in steady‐state conditions, have a low NADPH/NADP+ ratio, thus allowing the HMP to operate at its maximal intracellular rate and to compensate the intrinsic erythrocyte enzyme deficiency. Studies started soon after accidental intake of fava beans by sensitive G6PD‐deficient subjects demonstrate a decrease of both NADPH/NADP+ ratio and reduced glutathione. The metabolic effects induced by fava beans may be attributed to oxidative stress probably associated with an inhibitor effect of some unknown metabolite on the HMP. The availability of erythrocytes from subjects recovering from haemolysis with high reticulocyte counts and increased G6PD activity, provides new information on the rate of synthesis as well as on the in vivo decay of the mutant enzyme. Correlation of G6PD activity to reticulocyte count and extrapolation to an ideally homogenous population of reticulocytes reveal that the mutant enzyme is synthesized at a nearly normal rate. Furthermore, the present correlation allows an estimate of the in vivo half‐life of Mediterranean G6PD. The rate of decline of about 8 d observed in this study well correlates to the intracellular metabolic aspects of G6PD Mediterranean erythrocytes.
Platelets and Surface‐Mediated Clotting Activity
Authors: Vicic William J. , Ratnoff Oscar D. , Saito Hidehiko , Goldsmith George H.
Published: March 2008

Summary. In contrast to previously reported studies, no evidence could be adduced for the activation of Hageman factor (factor XII) by platelets, whether or not these cells had been incubated with adenosine diphosphate (ADP).
Cycles of Agglutination‐Disagglutination Induced by Ristocetin in Thrombasthenic Platelets
Authors: Chediak J. , Telfer M. C. , Laan B. Vander , Maxey B. , Cohen I.
Published: March 2008

Summary. An oscillatory pattern of platelet agglutination‐disagglutination in response to Ristocetin (R) at narrow concentration ranges was observed in citrated platelet rich plasma (PRP) of 10 patients with Glanzmann's thrombasthenia. The cyclic pattern decreased in intensity over time, was reproducible, and was not pH dependent. Formalin‐fixed thrombasthenic platelets agglutinated with R but did not show a cyclic pattern.

Incubation with 2.5 μm ADP inhibited R oscillation response, but small increases in R dose overcame this inhibition. The addition of ATP or creatine phosphate/creatine phosphokinase to thrombasthenic platelets inhibited by ADP restored the R oscillation response. In the platelets of a single patient, intracellular levels of ADP and ATP were shown to diminish during an oscillation response to R. There was an increase in AMP levels during the same period of time. The changes in these three intracellular nucleotides were gradual over time and did not vary with phases of the oscillation. Acetyl salicylic acid (ASA), at concentrations shown to block cyclooxy‐genase activity in control platelets, enabled thrombasthenic platelets to respond to R with full agglutination without oscillations. Lower concentrations of ASA in the PRP gave a return of the oscillation response.

Our data suggest that the disagglutination phase of the R response of thrombasthenic platelets is not a function of the known glycoprotein membrane defect, but depends on materials originating in the platelet whose release is blocked by ASA.
Storage Pool Disease: Comparative Fluorescence Microscopical, Cytochemical and Biochemical Studies on Amine‐Storing Organelles of Human Blood Platelets
Authors: Lorez H. P. , Richards J. G. , Prada M. Da , Picotti G. B. , Pareti F. I. , Capitanio A. , Mannucci P. M.
Published: March 2008

Summary. The platelet content of ATP, ADP, serotonin (5‐HT), dopamine, noradrenaline and adrenaline as well as the net uptake of radiolabelled 5‐HT and mepacrine were subnormal in six patients with Storage Pool Disease (SPD). Fewer amine‐storing organelles were found by fluorescence microscopy with the fluorescent probe mepacrine and by electron microscopy with a cytochemical (uranaffin) reaction specific for 5′‐phosphonucleotides. Both methods showed that SPD platelets have atypical organelles with a reduced capacity to store amines, 5′‐phosphonucleotides and mepacrine. The changes were most marked in platelets of the two patients who also had oculocutaneous albinism.
The Value of the Standardized Partial Thromboplastin Time in Detecting Heparin during Low‐Dose Prophylaxis
Authors: Taberner D. A. , Norman C. , Poller L. , Burslem R. W. , Jones J. B.
Published: March 2008

Summary. The sensitivity of the standardized partial thromboplastin time technique in detecting circulating heparin during low‐dose calcium heparin prophylaxis in gynaecological surgery has been evaluated and compared with an anti Xa assay and a chomogenic substrate method. The PTT test and anti Xa test were performed daily on plasmas from eight control and seven heparin treated patients. Both the PTT and anti Xa test showed significant prolongation in heparin‐treated patients receiving the 5000 units regime 12‐hourly. In control patients the PTT test showed a significant shortening, maximal on the fifth day after operation. This interferes with the detection of heparin by the PTT during this period. Nevertheless, the effect of small concentrations of heparin is observable during low‐dose prophylaxis by its prevention of the post‐operative acceleration of the PTT test. By contrast, the anti Xa test is relatively unaffected by the post‐operative acceleration and is more prolonged by the low‐dose heparin therapy.

The heparin levels in 17 plasmas from gynaecological surgical patients receiving low‐dose therapy were determined by PTT, anti‐Xa and chromogenic substrate. The chromogenic substrate and anti Xa assays showed reasonably good intra‐class correlation. Heparin levels determined by the PTT test were invalidated by the post‐operative acceleration of the test. If, however, this post‐operative acceleration is of clinical significance and relates to a prethrombotic state, its detection by the PTT may be important and may be a valuable indicator for the need for an increase of the heparin dosage. Unless it is shown that the anti Xa effect of heparin is of more importance in the prevention of thrombosis than the prolongation of the PTT, the standardized PTT appears to offer a simple method of control of low‐dose heparin administration.
Reduced Granulocyte‐Macrophage Colony‐Stimulating Activity by Mitogen‐Stimulated Lymphocytes from Patients with Aplastic Anaemia
Authors: Weatherly Thomas L. , Fleisher Thomas A. , Strong Douglas M.
Published: March 2008

Summary. The ability of peripheral blood mononuclear cells from patients with idiopathic aplastic anaemia to provide colony‐stimulating activity (CSA) was compared to that of normal controls. CSA was prepared by incubating peripheral blood mononuclear cells with phytohaemagglutinin. The supernatant derived from the latter is known as activated lymphocyte‐conditioned medium (ALCM). The CSA of ALCM in eight patients was compared to that of normals in 12 experiments. In all but one instance there was decreased CSA by patient ALCM. Possible implications of these findings are discussed.
Variability of Intracellular pH within Individual Populations of SS and AA Erythrocytes
Authors: Kaperonis A. A. , Bertles J. F. , Chien S.
Published: March 2008

Summary. Individual populations of AA and SS erythrocytes were fractionated according to cell density by centrifugation, and the fractions analysed for intracellular pH (pHi), the mole ratio of 2,3‐diphosphoglycerate to haemoglobin (DPG:Hb), and cell concentration of haemoglobin (MCHC). The pHi of SS erythrocytes was consistently lower than that of AA erythrocytes throughout the density range, and the lowest pHi of both cell types (AA and SS) was found in cells with the highest density. As the highest density AA and SS erythrocytes are characterized by the lowest DPG:Hb values, their relatively low pHi cannot be ascribed to intracellular organic phosphate. Instead we propose that a redistribution of hydrogen ions across the membranes of both AA and SS erythrocytes is the ultimate result of progressive alterations in these membranes in vivo.
Alpha Thalassaemia in Sicily: Haematological and Biosynthetic Studies
Authors: Musumeci Salvatore , Schiliro Gino , Pizzarelli Giuseppe , D'Agata Alfonsina , Fischer Alberto , Russo Giuseppe
Published: March 2008

Summary. Eight Sicilian patients with Hb H disease and their families have been studied. The standard haematological tests and the α/β chain synthesis ratios showed significantly different results in the patients with Hb H disease as compared with α thalassaemia carriers, except for Hb A2 values. There was no significant difference in the mean RBC, MCV, Hb A2, Hb A1 and Hb F of α thalassaemia carriers compared with normal controls. On the contrary significant difference was found between the mean α/β chain synthesis ratio of a thalassaemia carriers and that of the normal controls; however, the extensive overlapping of α/β values between these two conditions make this parameter insufficiently discriminant. No correlation was found between MCV, MCH, RBC and α/β chain synthesis ratio in patients with α thalassaemia trait, suggesting that the ratio cannot be used to distinguish between carriers of a mild gene (‘silent’carrier) and carriers of the more severe α thalassaemia gene. A possible genetic model for α thalassaemia in Sicily is presented.
Influence of Dietary Protein Concentration and Quality on Response to Erythropoietin in the Polycythaemic Rat
Authors: Alippi R. M. , Giglio J. M. , Barceló A. C. , Bozzini C. E. , Farina R. , Rio M. E.
Published: March 2008

Summary. The effects were examined of dietary protein concentration and quality on the response of polycythaemic hypertransfused rats to 6 units of human urinary erythropoietin. Rats were either starved or fed one of 14 different diets. Four protein sources were used, having a quality gradient from 100 to about 24. Two proteins—casein and wheat gluten—were used at five different levels of concentration (5–25%) in the diet. The response of rats maintained on the standard diet (Purina rat chow, 23.4% protein/g) was taken as the normal standard. The response to erythropoietin was 25% of normal in starved rats and 35% of normal in rats put on a protein‐free diet. When 10% protein in the diet was obtained by using high biological value proteins (egg yolk or casein) the response to erythropoietin was normal. When the same concentration was achieved by using low biological value proteins (wheat gluten or corn protein) the response to erythropoietin was undistinguishable from that of rats put on the protein‐free diet. When rats were maintained on diets with different concentrations of casein (5–25%) a normal response was observed when protein concentration was 10% with no further changes at higher concentrations. When rats were fed diets with different wheat gluten concentrations (5–25%) the response to erythropoietin was subnormal. These data suggest that the ability of rats to respond normally to erythropoietin is dependent on a continuous dietary intake of proteins at levels which are dependent on their biological values.
Cell Surface Heterogeneity of Human Blood Neutrophils and Monocytes
Authors: Barrett Sondra , Garratty Eileen , Garratty George
Published: March 2008

Summary. Until recently, human blood neutrophils (PMN) and monocytes have been considered to be homogeneous cell populations. However, much evidence has accumulated on their functional heterogeneity. This functional heterogeneity suggests the existence of different subsets of myeloid cells analogous to T and B subsets of lymphoid cells. The goal of this study was to investigate this question of myeloid subsets by examining myeloid cells for cell surface reactivity for IgG and complement (C).

Normal PMN and monocytes were examined from 60 subjects for the presence of two types of IgG‐Fc receptors and two activated C components, C3b and C3d. Most PMN and monocytes showed Fc receptor reactivity for rabbit IgG (Fc‐R). In addition, the majority of monocytes but very few PMN reacted with human IgG (anti‐Rh0) coated Rh‐positive erythrocytes (Fc‐H). Most PMN and monocytes showed C receptor reactivity for C3b, but only a minor subpopulation of both myeloid cells had C3d receptors. These data provide evidence that human blood myeloid cells may be composed of subsets with different membrane marker reactivities.
Idiopathic Neutropenia with Normocellular Bone Marrow: an Immune‐Complex Disease
Authors: Caligaris‐Cappio F. , Camussi G. , Gavosto F.
Published: March 2008

Summary. The presence of circulating immune‐complexes (IC) and their in vivo interaction with polymorphonuclear neutrophils (PMN) have been detected in two cases of idiopathic neutropenia with normocellular bone marrow. The injection of patients' sera into New Zealand White rabbits caused a striking neutropenia due to sequestration of PMN in the vascular bed of kidneys and lungs. The kinetics of PMN disappearance from peripheral blood and the pattern of sequestration overlapped that induced by the injection of pre‐formed soluble IC. Treatment with plasmapheresis caused an early and lasting increase of PMN; rabbit PMN were almost unaffected by the injection of patient serum after the course of plasmapheresis.

These data are consistent with the possibility that idiopathic neutropenia with normocellular bone marrow may be caused by persistent in vivo interaction between IC and circulating PMN.
The Chromatographic Separation of Factor VIII on Aminohexyl Sepharose
Authors: Austen D. E. G.
Published: March 2008

Summary. A new method of factor VIII purification has been devised which involves chromatographic separation on aminohexyl‐substituted agarose. Relatively large volumes of starting material can be processed compared to the volume of agarose employed and satisfactory yields are obtained. Factor‐VIII‐clotting activity is separated from the other related substances and resultant products appear to be stable. While separation of clotting activity occurs with relative ease, the antigen and ristocetin co‐factor are hardly segregated, if at all. This provides a little more information about the interrelation of these substances. Results suggest that ion‐exchange is involved in the mechanism of separation but additional hydrophobic or steric effects cannot be ruled out.
Properties of Anti‐Hairy Cell Serum
Authors: Stuart A. E. , Dewar A. E.
Published: March 2008

Summary. An antiserum was prepared in rabbits against hairy leukaemic cells. After absorption with human erythrocytes, thymus and liver powder, the serum was passed through immunoabsorbent columns to remove extensive anti‐plasma protein activity. Following these procedures, sensitive techniques failed to reveal residual activity against α‐macroglobulins, β2‐microglobulin and other human plasma proteins. The antiserum reacted with normal B lymphocytes but not with T lymphocytes or monocytes. Positive reactions were seen in B cell lymphomas, chronic lymphocytic leukaemia, hairy‐cell leukaemia and uninucleated Reed‐Sternberg cells.