bluuebunny/crossref_metadata_2025 huggingface dataset:
The Physiopathological Significance of Benign Monoclonal Gammopathy: a Study of 64 Cases
Authors: Carter Anna , Tatarsky Ilana
Published: March 2008

Summary.

Sixty‐four patients with monoclonal protein in serum but initially without evidence of multiple myeloma, macroglobulinaemia, amyloidosis or lymphoma, were studied. Fifty patients (78%) were observed for a period exceeding 3 years. Based on the follow‐up data the patients were classified into the following four groups: Group 1=patients with transient monoclonal gammopathy: 4.7%; Group 2 = patients without significant increase in monoclonal serum protein: 75%; Group 3 = patients with more than 50% increase in monoclonal serum protein: 14.1%; Group 4=patients in whom multiple myeloma developed: 6.2%. The mean interval from discovery of the serum monoclonal protein to evolution to multiple myeloma was 61 months. Retrospective analysis of age, sex, blood count, bone marrow picture, antigenic type and size of serum monoclonal proteins, presence of small amounts of homogeneous light chain in the urine, serum albumin level, levels of residual immunoglobulins, did not help to distinguish initially the patients in whom the monoclonal gammopathy evolved to multiple myeloma from patients in whom the disease remained benign and stable. The evolution to multiple myeloma had occurred abruptly after long periods of stable condition; and until this progression the follow‐up data were similar to the patients with benign disease. The possible physiopathology of occurrence and evolution of benign monoclonal gammopathy is discussed.
The Synthesis of Folate‐binding Protein in Lymphocytes during Transformation
Authors: Costa Maria da , Sharon Michael
Published: March 2008

Summary.

The blast transformation of human lymphocytes exposed to phytohaemagglutinin was associated with an increase in the intracellular concentration of a folate‐binding protein. The protein was measured by a radioinimunoassay and by immunofluorescent microscopy using an antiserum raised against purified folatebinding protein prepared from chronic myelogenous leukaemia cells. Because it has been previously shown that protein‐bound folate cannot function as a cofactor or substrate, the synthesis of this binder during some phase of the replicative cycle may serve to modulate folate‐dependent RNA and DNA synthesis by controlling the concentration of free folate.
Use of lllIndium‐labelled Platelets to Measure Spleen Function
Authors: Peters A. M. , Klonizakis I. , Lavender J. P. , Lewis S. M.
Published: March 2008

Summary.

The distribution of 111In‐labelled platelets following intravenous bolus injection has been studied using a gamma camera and computer system. Liver uptake, which accounted for about 10% of the dose, was completed between 6 and 10 min after injection. Blood pool and splenic 111In, which accounted for the remainder of the dose, reached constant levels simultaneously about 20 min after injection. The kinetics of splenic uptake are consistent with a two compartmental model in which circulating and splenic platelets are in dynamic equilibrium with each other. From analysis of the kinetics, splenic blood flow and the mean transit time of platelets through the spleen have been calculated in normal subjects and in patients with haematological disorders. Blood flow, which was about 200 ml per min in normals, tended to increase with increasing spleen size. Transit time was not dependent on spleen size; it was about 10 min in all but one of the subjects.
Radionuclide Distribution folowing Injection of 111Indium‐labelled Platelets
Authors: Klonizakis I. , Peters A. M. , Fitzpatrick M. L. , Kensett M. J. , Lewis S. M. , Lavender J. P.
Published: March 2008

Summary.

Platelets labelled with 111In displayed similar survival curves after incubation with 111In‐oxine in plasma, plasma‐saline and dextrose saline media. The use of autologous red cells to cushion platelets during high‐speed centrifugation facilitated platelet resuspension without greatly affecting the duration of the labelling procedure. Quantitative scanning after reinjection of labelled platelets in haematologically normal subjects showed that, initially, splenic indium amounted to about 35% of the injected dose and hepatic indium about 12%; these levels rose only slightly over the subsequent duration of the platelet life span. Subtraction of the signal from indium in platelets thought to be normally pooled within the spleen from the total indium signal gave splenic indium uptake curves which reflected splenic platelet destruction. Initially, the sum of indium levels in spleen, liver and blood equalled 100% of the dose. Thereafter, the sum fell progressively at a rate thought to be approximately equal to the rate of bone marrow uptake.
Biochemical and Immunological Characterization of Ferritin from Leukaemic Cells
Authors: Wagstaff M. , Worwood M. , Jacobs A.
Published: March 2008

Summary Ferritin was prepared from the leucocytes of four patients with acute myeloblastic leukaemia (AML) and eight patients with chronic myeloid leukaemia (CML). Isoelectric focusing demonstrated a prominent relatively basic band in all the CML cell ferritins. The iron content of all the ferritins was very low (max 180 atom/molecule). Thus there was little evidence for the role of leucocyte ferritin as an iron store or for iron as the major factor responsible for the initiation of ferritin synthesis. The iron uptake properties, subunit analyses, immunological reactivities and other immunological studies indicated that leukaemic cell ferritins had similar properties to tissue ferritins.
Quantitative Assessment of Erythropoiesis in Haemolytic Disease
Authors: Stefanelli M. , Barosi G. , Cazzola M. , Orlandi Ester
Published: March 2008

Summary Erythrokinetic parameters were estimated in 21 patients suffering from haemolytic disease using a mathematical model of iron kinetics. A proper definition of radioiron reflux from red cells to the plasma proved to be essential for an accurate estimate of effective and ineffective erythropoiesis in severe haemolytic conditions. Erythropoietic activity was increased in all the patients, with the marrow capable of sustaining erythropoiesis at about 6 times normal in hereditary spherocytosis and 3 times normal in autoimmune haemolytic anaemia. This proliferative effort was countered by an increase in ineffective erythropoiesis which was especially marked in hereditary spherocytosis.
Acidified Glycerol Lysis Test: a Screening Test for Spherocytosis
Authors: Zanella A. , Izzo C. , Rebulla P. , Zanuso F. , Perroni L. , Sirchia G.
Published: March 2008

Summary Acidified glycerol lysis test (AGLT) is a screening procedure which has been developed for spherocytosis. AGLT was found positive in 100% of 48 patients suffering from hereditary spherocytosis, 100% of nine couples of affected parents, and 86% of 14 couples of clinically healthy parents. The test was positive in acquired spherocytosis and negative in normal controls. AGLT appears to have a predictive value higher than the entire battery of conventional tests. It is simple, rapid, inexpensive, gives clear‐cut results and requires minute amounts of blood. It can be performed on blood stored for up to 24 h.
Isolation and Characterization of Normal Human Megakaryocytes
Authors: Levine R. F.
Published: March 2008

Summary Human megakaryocytes have been isolated from marrow obtained from ribs removed at thoracotomy. All but one of the patients had normal pre‐operative platelet and leucocyte counts. Megakaryocytes averaged 0·37% of all cells in marrow cell suspensions from nine consecutive subjects. A 283‐fold purification (to 10·3%) was achieved by a density gradient centrifugation followed by two successive velocity sedimentations at unit gravity. The net yield, 12 800 megakaryocytes per specimen, was sufficient for many kinds of morphological study. Bright‐field, phase contrast, and electron microscopy were used to characterize the younger and smaller megakaryocytes. Ploidy analyses were carried out on 100–235 megakaryocytes per specimen; 8N was the predominant ploidy class in isolated megakaryocyte populations from three individuals. The mean megakaryocyte diameter was 24 μm in three other specimens and the range was 10–48 μm. This data had a normal distribution and overlapped minimally with the size range of all other marrow cells. The presence of a distinct size threshold (at 11·5 μm) implied that size alone may be a sufficient objective criterion for identification of human megakaryocytes.
Factor VIII and Fibrinolytic Response to Deamino‐8‐d‐Argenine Vasopressin in Normal Subjects and Dissociate Response in Some Patients with Haemophilia and von Willebrand's Disease
Authors: Ludlam C. A. , Peake I. R. , Allen N. , Davies B. L. , Furlong R. A. , Bloom A. L.
Published: March 2008

Summary Deamino‐8‐d‐argenine vasopressin (DDAVP) was given by intravenous infusion to normal subjects, haemophiliacs and patients with von Willebrand's disease (vWd) and the factor VIII and plasminogen activator response was studied. In normal subjects and most patients with mild haemophilia and mild (intermediate) von Willebrand's disease there was an increase in plasminogen activator and all factor VIII related activities. In patients with mild vWd the prolonged bleeding time was shortened by DDAVP despite only a modest rise in factor VIII related Ristocetin cofactor activity (VIIIR:RiCoF). Sub‐groups of patients have been characterized in whom atypical responses were observed. In two brothers with clinically severe haemophilia, but with 5–6 u/dl procoagulant factor VIII (VIIIC), there was an increase in VIIIC but no rise of the corresponding antigen, suggesting increased release of an antigenically abnormal poorly functioning molecule. A patient with intermediate vWd was studied in whom neither DDAVP, adrenaline infusion, nor venous occlusion resulted in an increase in either plasminogen activator or factor VIII related antigen (VIIIRAg), although there was a significant increase in VIIIC. In a further patient with severe vWd, DDAVP failed to elicit any plasminogen activator or VIII response. The results obtained from these two patients suggested that in some individuals the presumed endothelial cell abnormality in vWd may be more extensive than a defect in VIIIRAg synthesis. Sub‐groups of patients have been identified for whom treatment with factor VIII concentrates would be more appropriate than DDAVP prior to minor surgery.
Globin Chain Electrophoresis: a New Approach to the Determination of the Gγ/Aγ Ratio in Fetal Haemoglobin and to Studies of Globin Synthesis
Authors: Alter Blanche P. , Goff Sabra C. , Efremov Georgi D. , Gravely Marsha E. , Huisman Trrus H. J.
Published: March 2008

Summary. Separation of globin chains by electrophoresis provides a simple and rapid method for the determination of the Gγ/Aγ ratio in human fetal haemoglobin, and of biosynthetic rates of the globin chains. Whole haemolysates were analysed by electrophoresis on polyacrylamide gels in urea, acetic acid and Triton X‐100. Electrophoresis of haemolysates from newborn infants led to four bands: Aγ, Gγ, β and α. The identity of these bands was indicated by examination of haemoglobins of known globin chain composition. In 15 samples, the %Gγ was similar by Triton gels and by amino acid analysis of the γCB‐3 peptide. Some mutant globin chains were also separable with the electrophoretic technique. Triton gel electrophoresis provides rapid analysis of very small amounts of haemoglobin, and permits examination of globin chain composition as well as globin synthetic ratios.
Fetal Hb Production during Acute Erythroid Expansion: I. OBSERVATIONS IN PATIENTS WITH TRANSIENT ERYTHROBLASTOPENIA AND POST‐PHLEBOTOMY
Authors: Papayannopoulou Th. , Vichinsky E. , Stamatoyannopoulos G.
Published: March 2008

Summary. In order to study fetal haemoglobin production during acute erythroid expansion we did sequential measurements of Hb F‐containing erythrocytes (F‐cells) and of relevant haematological parameters in 10 subjects recovering from erythroid aplasia, iron deficiency anaemia or following phlebotomy. An increased production of F‐cells was consistently observed during the acute marrow expansion, but there were significant differences in the maximum F‐cell response among individuals. These differences could not be explained by differences in the degree of anaemia alone, nor could they be correlated with the level of peak reticulocytosis. Two patients who reached the highest F‐cell numbers were probably carriers of heterocellular hereditary persistence of Hb F, suggesting that this gene may play a role in determining the magnitude of F‐cell production in anaemic patients.

It is speculated that the main reason for the consistently observed increase in F cell production during acute marrow expansion is the premature terminal differentiation of earlier erythroid precursors (burst forming units; BFUe). This proposition is in accord with observations in vitro which suggest that Hb F is expressed in erythroid clones derived from BFUe's. It is further proposed that differences in the degree of BFUe recruitment may underly the disparities in Hb F response among individuals subjected to similar anaemic stimuli.
An Identical Translocation between Chromosome 1 and 7 in Three Patients with Myelofibrosis and Myeloid Metaplasia
Authors: Geraedts J. P. M. , Ottolander G. J. den , Ploem J. E. , Muntinghe O. G.
Published: March 2008

Summary. An identical chromosome abnormality was observed in three unrelated patients with myelofibrosis and myeloid metaplasia, two of the patients showing a history of polycythaemia vera (PV) before development of the myelofibrosis. Unstimulated peripheral blood cultures showed a translocation between chromosomes 1 and 7 replacing a homologue of pair 7. It was identified by G‐ and C‐banding as t(1;7)(7pter→7p11::1p1?→1qter).

While the first patient also showed trisomy 21 and the third patient had some extra material on the short arm of chromosome 17, all three had trisomy 1q and monosomy 7q. Although each of these abnormalities is frequently observed separately in various haematological disorders, the combination of the two in the form of an identical translocation in three patients is an example of induced non‐random cytogenetic change in myelofibrosis.
Studies on the Concentration and Intracellular Localization of Iron Proteins in Liver Biopsy Specimens from Patients with Iron Overload with Special Reference to their Role in Lysosomal Disruption
Authors: Selden Clare , Owen Marie , Hopkins J. M. P. , Peters T. J.
Published: March 2008

Summary. Liver biopsies were collected from control subjects and patients with iron overload due to either primary or secondary haemochromatosis. They were analysed for iron proteins by cation exchange chromatography and flameless atomic absorption spectrophotometry.

In control tissue the transferrin fraction contains 25%, ferritin 50% and haemprotein and haemosiderin 10–15% each, of the total iron. In iron overloaded tissue the ferritin and haemosiderin iron increases approximately 10‐ and 100‐fold, respectively, compared with control tissue.

There was a close positive correlation between enhanced lysosomal fragility as determined by measurements of latent N‐acetyl‐β‐glucosaminidase and haemosiderin content of the tissue; it is suggested that the haemosiderin is responsible for the lysosomal disruption and hence the tissue damage in iron overload.

Studies were performed on the intracellular localization of ferritin and of total iron in biopsy extracts from control subjects and from patients with iron overload. In control tissue, ferritin contains most of the iron and is apparently free in the cytosol. In iron overload, ferritin is the major iron protein in the post‐nuclear supernatant sedimenting into the gradient as the free protein. There are, however, significant amounts of immunoreactive ferritin deeper in the gradients but this cannot be assigned to any particular subcellular organelle. The extreme fragility of lysosomes in iron overloaded human tissue makes isolation of these organelles for detailed biochemical analysis extremely difficult.
Frozen Red Cells in Rhesus Immunization
Authors: Cook I. A. , Robb Anne L. , Mitchell R. , McLaren Elizabeth A. , Urbaniak S. , Robertson Anne E.
Published: March 2008

Summary. The use of frozen washed cells in varying doses in primary Rh immunization is compared in two groups of men with the use of fresh washed cells in a third group. In the first two groups, using frozen cells, doses ranging from 0.5 to 20 ml of whole blood (Group I) are compared with a 200.0 ml dose of red cell concentrate (Group II), while Group III served as a control using a 20 ml dose of fresh washed red cell suspension (9.0 ml concentrated red cell equivalent). The response rate was 93% in Group II compared with only 43% in Group I, suggesting the desirability of using relatively large doses of Rh‐positive red cells for primary Rh immunization. The use of frozen washed cells from a special panel for ‘booster’ injections is also recommended.
Effect of Different Resuspension Media on the Post‐thaw Characteristics of Frozen Blood
Authors: Amer Kamal A. , Pepper Duncan S. , Prowse Christopher V.
Published: March 2008

Summary. Red blood cells frozen by the low‐glycerol fast‐freezing technique were thawed, deglycerolized and resuspended in various media. The use of ACD‐saline for resuspension markedly reduced in vitro haemolysis such that the red cells could be transfused up to 5 d after thawing. At this time the cells contained satisfactory levels of potassium and organic phosphates, while bacterial contamination was negligible. For the past 4 years we have operated a small bank of previously frozen red blood cells which were thawed and resuspended in this way. Over 3500 units have been transfused after 1–5 d post‐thaw storage without untoward effects.
Identifying Carriers of Mild Haemophilia
Authors: Graham John B. , Barrow Emily S. , Flyer Paul , Dawson Deborah V. , Elston R. C.
Published: March 2008

Summary. The problems of carrier identification in mild haemophilia were examined by studying a large kindred transmitting VIII:C levels which averaged 17 u/dl in affected males. Fifteen obligatory carriers and 13 normal women from this kindred were used as reference groups to produce a set of linear discriminants. The utility of these discriminants in identifying carriers of mild haemophilia was compared with that of a similar set of discriminants which had been prepared for use with carriers of severe haemophilia, each set of discriminants being tested against both data bases. It was found that both sets of discriminants had large error rates when applied to the mild data base, and that both had much smaller error rates when applied to the severe data base. This outcome resulted from the greater overlap between the factor VIII‐related activities of mild carriers and normal women than the overlap of these activities between severe carriers and normal women.

Our conclusions were that (i) correct classification of many but not all women from families of mild haemophilia would be difficult, (ii) discriminants prepared using obligatory carriers of severe haemophilia were not more inefficient in mild haemophilia than those prepared using obligatory carriers of mild haemophilia, (iii) a single set of discriminants prepared from reference groups of normal women and obligatory carriers of severe haemophilia may be used for identifying potential carriers of all grades of haemophilia, and (iv), statistically speaking, VIIIR:Ag assays provide a limited amount of information for this purpose.
Quantitation of Binding of Factor VIII Antigen to Concanavalin A
Authors: Howard Margaret A. , Perkin Jean , Koutts Jerry , Firkin Barry G.
Published: March 2008

Summary. This study establishes a convenient method for screening plasma samples for abnormalities of the carbohydrate content of the factor VIII (FVIII) molecule. A radioimmuno‐electrophoretic technique has been developed to quantitate the percentage binding of FVIII‐related antigen (VIII‐Ag) to the lectin concanavalin A (Con A). Plasma samples were electrophoresed through a strip of agarose containing Con A into agarose containing a mixture of unlabelled anti‐FVIII and 125I‐anti‐FVIII where precipitant lines formed, the height of which was dependent upon the degree of VIII‐Ag binding to Con A in the first gel. Using this system reduced binding of VIII‐Ag to Con A was found in the plasma of 12 patients with moderate classical von Willebrand's disease (vWd), while the Con A binding of six haemophilia A patients fell within the normal range. The VIII‐Ag in normal cryoprecipitate showed increased % binding to Con A while the VIII‐Ag remaining in the cryo‐supernate demonstrated reduced Con A % binding.
Synthesis and Release of Factor VIII by Cultured Human Endothelial Cells
Authors: Tuddenham Edward G. D. , Lazarchick John , Hoyer Leon W.
Published: March 2008

Summary. Endothelial cells (ECs) derived from human umbilical veins were cultured in order to study the physiological control of factor VIII synthesis and release. The culture media were studied from multiple replicate cultures at confluence. Factor VIII related antigen (VIIIR:Ag) and factor VIII coagulant antigen (VIII:CAg) were measured by sensitive immunoradiometric assays. De novo synthesis of factor VIII related protein (VIII:R) was quantitated by incorporation of labelled amino acids into specific protein subunits. The following agents were added to the culture medium in a range of concentrations from physiological to pharmacological: adrenaline, 5 hydroxytryptamine, 2,3‐DPG, cyclic AMP, thyroxine, hydrocortisone, and human growth hormone. None of them had any effect at any concentration on the rate of accumulation of VIIIR:Ag in the culture medium. Addition of exogenous factor VIII had no effect on do novo synthesis of VIII:R. VIII:CAg was found to be stable under the conditions of culture but none was released from the ECs. Long‐term monocyte cultures also failed to release VIII:CAg. It appears that VIII:R is a constitutive gene product of umbilical vein endothelial cells and that VIII:CAg is not made by these cells.
Haemoglobin Synthesis in Human Erythroid Bursts during Ontogeny: Reproducibility and Sensitivity to Culture Conditions
Authors: Darbre Philippa D. , Lauckner S. M. , Adamson J. W. , Wood W. G. , Weatherall D. J.
Published: March 2008

Summary. Patterns of haemoglobin (Hb) synthesis were analysed in erythroid bursts grown in vitro from human fetal, umbilical cord and adult blood to determine the reproducibility of the system and its sensitivity to various alterations in culture conditions. The proportion of Hb F synthesis in duplicate plates from the same sample showed little variation but when an individual was studied under the same conditions but at different times, far more variability was observed.

We have confirmed that the synthesis of Hb F and Hb A in the cultures is asynchronous, relatively more Hb F being synthesized earlier in the maturation of the colony. Hence factors affecting colony maturation are likely to change the apparent pattern of Hb synthesis. Removal of adherent cells prior to plating resulted in the presence of more mature colonies and lower levels of Hb F synthesis at any fixed time in culture. Furthermore, the overall ratio of γ:β chain synthesis measured in the whole plate was dependent on the proportion of large, immature multicluster bursts present. The significance of measurements of Hb synthesis in erythroid bursts can be assessed only if all of these factors are recognized and taken into account.
Ichthyosis and Marrow Involvement in Malignant Histiocytosis of the Intestine
Authors: McCann S. R. , Barry D. P. , Temperley I. J. , Weir D. G.
Published: March 2008

Summary. It has recently been suggested that a number of small bowel lymphomas arise from histiocytic cells in the intestine. This has been referred to as malignant histiocytosis of the intestine (MHI) (Isaacson et al, 1979). Three such patients with small bowel malabsorption had the following features: subtotal villous atrophy of the jejunum, generalized ichthyosis, fever and lymphopenia are described. In all three, the bone marrow appearances were similar to those seen in histiocytic medullary reticulosis (HMR) (Scott & Robb‐Smith, 1939). A tumour was present in the small bowel and/or mesentery of all patients and the histological lesion was similar to that described as MHI. Two patients had a response to combination chemotherapy. One patient had a complete remission of his disease but this relapsed after 1 year and proved refractory to chemotherapy. A second patient died following chemotherapy with gastro‐intestinal perforation and septicaemia and the third patient died shortly after diagnosis and before chemotherapy could be commenced. It is suggested that the combination of signs, symptoms and pathological features described may reflect a specific clinical entity which has not previously been described.
Thrombin‐induced Proteolysis of Human Antithrombin III: an Outstanding Contribution of Heparin
Authors: Marciniak Ewa
Published: March 2008

Summary. Products obtained in reaction of excess antithrombin III (AT III) with human α‐thrombin and heparin were found to contain markedly less of residual AT III than products of similar reactions without heparin. The increased utilization of AT III was primarily due to limited proteolysis of a portion of unbound inhibitor, associated with the release of a 50 000‐dalton protein fragment. Thrombin‐induced release of this fragment was promoted by polydispersed heparin preparation and, to a variable degree, by all heparin fractions obtained in gel filtration. The optimum amount of heparin required to facilitate AT III proteolysis was of 5 μg/ml (0.8 u/ml). Excessive reduction of the residual inhibitory activity and changes in two‐dimensional immunoelectrophoresis suggested that AT III in plasma is also subjected to nonproductive proteolysis and formation of a modified AT III derivative following addition of thrombin and heparin. These data indicate that the effect of heparin on AT III is more complex than generally recognized. On the one hand, heparin interacting with AT III and thrombin contributes to a rapid binding and neutralization of the enzyme; on the other, heparin facilitates proteolytic degradation of unbound inhibitor even in the presence of small quantities of thrombin accounting for excessive reduction of the overall inhibitory potential of AT III.
CALCULATING THE DOSE OF FACTOR VIII IN THE MANAGEMENT OF HAEMOPHILIA
Authors: Ingram G. I. C.
Published: March 2008

Summary. Three methods of calculating the dose of factor VIII have been compared. A simple formula based on body weight has been found satisfactory for cryoprecipitate and freeze‐dried concentrate; but where body build departs markedly from average, height should probably also be taken into account. Modifications for children are noted.
Transient Donath‐Landsteiner Haemolytic Anaemia
Authors: Wolach Baruch , Heddle Nancy , Barr Ronald D. , Zipursky Alvin , Pai K. R. Mohan , Blajchman Morris A.
Published: March 2008

Summary. Five children with ‘paroxysmal cold haemoglobinuria’ are described. In all of them an upper respiratory tract infection preceded the episode of acute intravascular haemolysis and potentially pathogenic organisms were recovered from four. The haematological features are presented and the importance of technique in the detection of the pathognomonic antibody is stressed and discussed in detail. Although the clinical manifestations are usually self‐limiting and of short duration, it may be necessary, because of the rapidity and severity of anaemia, to provide interim support with appropriate blood transfusion. Since the disease is not usually recurrent and not clearly related to exposure to cold, it is suggested that it be re‐named Donath‐Landsteiner haemolytic anaemia.
Bernard‐Soulier Disease: a Study of Four Patients and their Parents
Authors: George James N. , Reimann Terry A. , Moake Joel L. , Morgan Rebecca K. , Cimo Phillip L. , Sears David A.
Published: March 2008

Summary. Two families with Bernard‐Soulier disease, including four patients and three of their parents, were studied and detailed clinical summaries are presented. One patient in each family has suffered severe bleeding problems while the other affected sibling is less severely affected. There has been no excessive bleeding in any of the parents or other family members. The patients demonstrated the abnormalities characteristic for Bernard‐Soulier disease: thrombocytopenia, giant platelets, prolonged bleeding time, abnormal platelet aggregation to human FVIIIvWF and ristocetin or bovine FVIIvWF alone, defective ristocetin‐induced binding of human 125I–FVIIIvWF multimers, decreased platelet lysis by a drug‐dependent antibody and complement, and a decreased concentration of membrane glycoprotein I. The parents had normal platelet counts, bleeding times, and FVIII‐mediated aggregation. However, the parents had anormally large platelets, decreased sensitivity to lysis by a drug‐dependent antibody and complement, and a decreased concentration of membrane glycoprotein I. Therefore the heterozygous state for Bernard‐Soulier disease is recognizable by platelet membrane abnormalities although there is no defect of platelet function and no excessive bleeding. Red cell membrane proteins of one patient were normal, suggesting that phenotypic expression of the Bernard‐Soulier disease defect is restricted to platelets.
Interaction between Homozygous β0 Thalassaemia and the Swiss Type of Hereditary Persistence of Fetal Haemoglobin
Authors: Cappellini M. D. , Fiorelli G. , Bernini L. F.
Published: March 2008

Summary.

The interaction between β0 thalassaemia and an heterocellular form of hereditary persistence of fetal haemoglobin (HPFH), presumably of the Swiss type, has been studied in three generations of a family in which both traits occur. The haematological parameters and the segregation of the two characters in the family suggest that the propositus, a 52‐year‐old male from southern Sardinia, is homozygous for β0 thalassaemia and carrier of the HPFH.

In spite of the complete suppression of adult haemoglobin synthesis, the patient is not anaemic and shows only morphological abnormalities of the red cells associated with a moderate decrease of the erythrocyte life span. Studies of the synthesis of haemoglobin chains in vitro have revealed only a mild degree of unbalance in the propositus, with a γ/α ratio of 0–67, and a very slight unbalance in a 3‐year‐old child heterozygous for β thalassaemia and HPFH.

Preliminary analysis of the linkage between this kind of heterocellular HPFH and the β Hb locus has been performed, utilizing all the suitable families reported in the literature. Although positive lod scores (1.535) have been obtained at a recombination fraction of 0.20, the data available are not sufficient to conclude in favour or against the linkage between the β Hb locus and the heterocellular type of HPFH.
Globin Biosynthesis in Erythroid Bursts of Heterozygous α or β Thalassaemia
Authors: Steinberg Martin H. , Coleman Mary B. , Pressley Annette , Adams Junius G.
Published: March 2008

Summary. We examined globin chain synthesis in erythroid bursts (BFU‐E) of patients with heterozygous α or β thalassaemia. BFU‐E were cloned from circulating mononuclear cells, labelled with [H]leucine and globin chains purified by gel filtration and column chromatography. In six patients heterozygous for β thalassaemia, globin synthesis in BFU‐E was nearly balanced, with an α/non α ratio of 1.05 0α12. These BFU‐E produced 33.8 12.7%γ globin chain, an amount similar (P >1 0.05) to that found in 10 controls with sickle cell anaemia (25.6 6.7) but greater (P <0.05) than that of five normal controls (17.2 2.2). The balanced globin synthesis appeared due to the large amounts of γ chain made by BFU‐E. In two α thalassaemia carriers, who also had sickle cell trait, the BFU‐E α/non‐α ratio was 0.67 and 0.79. These BFU‐E produced 15% and 20%γ chain and 39% and 45%βS globin. The synthesis of βS globin in BFU‐E exceeded the erythrocyte levels of 20% and 29% HbS and indicated nearly equal expression of βA and βB globin genes in these proliferating erythroid precursors. This provides further evidence that the low levels of HbS in sickle cell carriers with α thalassaemia are due to post‐translational events resulting from the differing affinity of βS and βA globin for α chain and the destruction of excessive βS chain.
Staging of Chronic Myeloid Leukaemia
Authors: Tura Sante , Baccarani Michele , Corbelli Giovanna
Published: March 2008

Summary. The prognostic value of nine clinical and haematological features, recorded at diagnosis in chronic myeloid leukaemia (CML), was analysed in two distinct series of patients. One series (116 cases) was collected at a single hospital over a 12‐year period. The second series (139 cases) was collected from a multicentre trial over a 20‐month period. Six features were associated with a poor prognosis: splenomegaly (more than 15 cm below the costal margin), hepatomegaly (more than 6 cm below the costal margin), thrombocytopenia (< 150 109/l) or thrombo‐cytosis (>500 109/l), a leucocyte count above 100 109/1, peripheral blood non‐granulated precursors (blast cells) above 1%, and peripheral blood granulated precursors (promyelocytes and myclocytcs) above 20%. Depending on the number of negative prognostic factors, patients were divided into three categories: group I (0 or 1 factor), group II (2 or 3 factors) and group III (4, 5 or 6 factors). Survival was significantly different in the three groups (P < 0.0005), and this was independent of age (below and above 50). The prognostic value of the classification was confirmed in a third scries of 153 patients. We suggest that this classification provides a useful tool to identify prognostic categories in CML, and thus allows a proper allocation of patients to different therapies.
Membrane Proteins Synthesized by Human Reticulocytes and their Precursors
Authors: Krasnow S. H. , Pielichowski H. J. , Caro J. , Burka E. R. , Ballas S. K.
Published: March 2008

Summary. Membrane protein synthesis in human immature erythroid cells was studied by incubating the cells with 5S‐methionine in vitro. The radioactive precursor amino acid was incorporated into membrane protein in a linear fashion for approximately 60 min, after which there was only a slight increase in incorporation. Intracellular protein synthesis, in contrast, was linear for periods up to 2 h. Analysis of isolated membranes by polyacrylamide gel electrophoresis in sodium dodecyl sulphate showed that peripheral blood reticulocytes synthesized two proteins in the 4.5 region (MW = 50–60 000 D) and a third protein coinciding with band 6 (glycer‐aldehyde‐3‐phosphate dehydrogenase). Separation of reticulocytes into different age groups on stractan II gradients showed more immature reticulocytes synthesize a wider range of membrane proteins, extending from bands 4.1 to 8. When nucleated red cells were present in the incubations, synthesis of band 3 was also observed. Earlier erythroid precursor present in erythropoietic BFU‐E cultures synthesized spectrin in addition to the other membrane proteins. The data indicate that human red cell membrane protein synthesis follows a programmed pattern and that as the erythroid elements mature they lose the capacity to synthesize certain membrane proteins.
Hereditary Bleeding Disorder due to a Primary Defect in Platelet Release Reaction
Authors: Wu Kenneth K. , Minkoff I. M. , Rossi Ennio C. , Chen Yao‐Chang
Published: March 2008

Summary. A large family with a hereditary bleeding disorder was investigated. Easy bruising, epistaxis and menorrhagia were noted in seven members of three generations and at least one member in each generation was affected. Platelet function abnormalities were characterized by reduced 14C‐serotonin release, absent second wave aggregation in response to ADP or epinephrine and reduced aggregation in response to collagen. Bleeding time was prolonged in three individuals and platelet factor 3 availability was abnormal in four. Platelet count, morphology, adhesiveness and clot retraction were normal in all. Platelet ADP and ATP as well as ATP to ADP ratio were normal. This family probably represents the first documented instance of hereditary platelet primary release disorder. To elucidate the pathogenetic mechanism, further functional studies were performed. No appreciable shape change, 14C‐serotonin release of aggregation was observed when the propositus’ platelets were stimulated with sodium arachidonate or a PGH2 analogue. By contrast, platelets responded normally to ionophore A23187, thrombin and ristocetin. The findings indicate that the hereditary primary release disorder is probably due to a reduced thromboxane A2 production secondary to thromboxane synthetase deficiency. Alternatively, it may be due to platelet membrane abnormalities which render platelets unresponsive to thromboxane A2.
Three Pyruvate Kinase Variants with Increased Affinity for PEP
Authors: Elder G. Elizabeth , Lappin T. R. J. , Lawson Barbara E. , Bridges J. M.
Published: March 2008

Summary. Three variants of pyruvate kinase are described which have marked reduction of activity associated with severe non‐spherocytic haemolytic anaemia. Each variant shows a reduced K0.5 PEP (the value of the intercept of the abscissa on the Hill plot) and reduced Hill coefficient; FDP activation and ATP inhibition are less than normal and utilization of GDP is increased. The variants are slightly less inhibited by 2,3DPG than controls but require more FDP to relieve this inhibition. Cases 1 and 2 have decreased thermostability but case 3 is normal. The mutant enzymes are further distinguished by their affinity for FDP. Their kinetic and physicochemical properties are compared with other known cases with high affinity for PEP and discussed in terms of a RT model for allosteric enzymes.
Regulation of Plasma Ferritin by the Isolated Perfused Rat Liver
Authors: Mack U. , Cooksley W. G. E. , Ferris R. A. , Powell L. W. , Halliday J. W.
Published: March 2008

Summary.The isolated perfused rat liver has been used to investigate the regulation of plasma ferritin in normal, iron‐deficient and iron‐overloaded states. Both125I‐labelled and non‐labelled rat liver ferritins were rapidly cleared from the perfusion circuit with a half‐life of approximately 30 min. Perfusion of livers from normal, iron‐loaded or iron‐deficient rats with blood obtained from normal, iron‐loaded or iron‐deficient rats showed that the liver takes up plasma ferritin, and releases ferritin into the perfusate to achieve a perfusate ferritin level appropriate to the iron stores of the animal from which the liver was taken. It is concluded that the liver is capable of both uptake and release of ferritin and that within the liver there resides a mechanism which maintains circulating ferritin concentrations at a level appropriate to body iron stores.
Suppression of in Vitro Granulocyte‐Macrophage Colony Formation by the Peripheral Mononuclear Phagocytic Cells of Patients with Idiopathic Aplastic Anaemia
Authors: Suda Toshio , Mizoguchi Hideaki , Miura Yasusada , Kubota Kazuo , Takaku Fumimaro
Published: March 2008

Summary. To investigate whether phagocytic cells play a role in the pathogenesis of aplastic anaemia, we studied the effect of the mononuclear phagocytic cells in the peripheral blood from patients with idiopathic aplastic anaemia on granulocyte—macrophage colony (GM‐colony) formation. There was no apparent infection in the studied patients. In nine out of 20 cases the mononuclear cells suppressed the formation of GM‐colonies by normal bone marrow. Removal of the phagocytic cells by carbonyl iron powder caused a loss of this suppressive effect. These results suggest the requirement for phagocytic cells in this suppressive effect and the possible involvement of phagocyte‐mediated immunological mechanism in the pathogenesis of some cases of idiopathic aplastic anaemia.
Induction by Phorbol Esters of Macrophage Differentiation in Human Leukaemia Cell Lines does not Require Cell Division
Authors: Territo Mary C. , Koeffler H. Phillip
Published: March 2008

Summary. Phorbol esters can induce differentiation into macrophage‐like cells in the two human myeloid leukaemia cell lines HL‐60 and KG‐.1. We investigated the requirement for DNA synthesis in this differentiation process. We found that phorbol ester‐induced differentiation of blast cells into cells with the morphologic, histochemical, and phagocytic characteristics of macrophages did not require H‐thymidine incorporation. Our results indicate that phorbol ester induced macro‐phage differentiation does not require DNA synthesis.
Studies on the Biochemical Sequelae of Therapy in Thy‐Acute Lymphoblastic Leukaemia with the Adenosine Deaminase Inhibitor 2’Deoxycoformycin
Authors: Russell N. H. , Prentice H. G. , Lee N. , Piga A. , Ganeshaguru K. , Smyth J. F. , Hoffbrand A. V.
Published: March 2008

Summary. In four patients with Thy‐acute lymphoblastic leukaemia changes in blast cell deoxynucleoside triphosphate concentrations and, in three, changes in blast cell S‐adenosyl homocysteine hydrolase activity were measured during treatment with 2’deoxycoformycin, a potent inhibitor of adenosine deaminase. These studies were aimed at identifying the molecular basis of cell killing by this drug. In three patients an increase in blast deoxyadenosine triphosphate (dATP) concentration occurred which was found to be temporally related to cell killing and was accompanied by decreased concentrations of the other three deoxyribonucleoside triphosphates. In the one patient with Thy‐ALL who responded poorly to treatment, the increase in dATP concentration was delayed and was not accompanied by a fall in the concentrations of the other deoxyribonucleoside triphosphates. Progressive inactivation of blast cell S‐adenosyl homocysteine hydrolase was found to occur in the three patients tested but was maximal only after a substantial reduction of peripheral blast cell count. These results show that 2’deoxycoformycin has a potent cytoreductive effect in Thy‐ALL and suggest that the molecular basis of this toxicity is related both to the intracellular accumulation of dATP with inhibition of ribonucleotide reductase. Inactivation of S‐adenosyl homocysteine hydrolase may be of importance as an additional mechanism.
Idiopathic Myelofibrosis: a Possible Role for Immune‐Complexes in the Pathogenesis of Bone Marrow Fibrosis
Authors: Cappio F. Caligaris , Vigliani R. , Novarino A. , Camussi† G. , Campana D. , Gavosto F.
Published: March 2008

Summary. Sixteen patients with idiopathic myelofibrosis (IM) have been investigated with respect to the possibility that immune mechanisms may be of importance in the pathogenesis of bone marrow fibrosis. The following points appear relevant: (1) immune‐complexes (IC) are detectable with different techniques in a high percentage of patients with IM. Their presence is associated with evidence of bone‐marrow histological markers of immune activity. (2) IgG is the main Ig class in the composition of IM IC. The results obtained favour the hypothesis that autoimmune mechanisms are involved in IM patients.
Entry of Malaria Parasites into Resealed Ghosts of Human and Simian Erythrocytes
Authors: Dluzewski A. R. , Rangachari K. , Wilson R. J. M. , Gratzer W. B.
Published: March 2008

Summary. Whereas‘white’ghosts, conventionally prepared by hypotonic lysis and washing are resistant to invasion by malaria parasites, cells lysed at high haematocrit and resealed without separating the membranes from the cytosol are invaded with high efficiency (not less than one‐third the frequency of intact cells). Similar results were obtained with human and monkey cells, exposed respectively to Plasmodium falciparum and P. knowlesi. Microspectrophotometry of individual cells was used to ascertain that the susceptibility to invasion of lysed, resealed cells was not related to its haemoglobin content.
Unstable Haemoglobin Haemolytic Crises: Contributions of Pyrexia and Neutrophil Oxidants
Authors: Winterbourn Christine C. , Williamson David , Vissers Margret C. M. , Carrell Robin W.
Published: March 2008

Summary. Pyrexia and the production of oxidants by phagocytic cells have been examined as two possible causes of haemolytic crises associated with infections in carriers of unstable haemoglobins. Three unstable haemoglobins were examined, both in red cells and after purification. Incubation at 40°C rather than 37°C resulted in only a slight increase in autoxidation rate, but a considerable increase in the rate of precipitation of the haemoglobins as Heinz bodies. Oxidants produced by activated neutrophils were capable of oxidizing haemoglobin both in solution and red cells, though there was no preferential effect on the unstable as opposed to the normal haemoglobin.

It is concluded that haemolytic crises associated with infections in carriers of unstable haemoglobins can be explained by increased intracellular precipitation of the haemoglobin as Heinz bodies caused by the accompanying pyrexia.
Serum Ferritin in Patients with Hereditary Spherocytosis
Authors: Blacklock Hilary A. , Meerkin Matthew
Published: March 2008

Summary. Serum ferritin was measured in 61 patients with autosomal dominant hereditary spherocytosis (HS), 44 splenectomized and 17 with intact spleens. In the majority (78%) the serum ferritin concentration was not elevated. Thirteen (22%) had mildly elevated levels, including five splenectomized females (range 181‐236 μg/l), and eight males, two with intact spleens (range 236‐436 μg/l). The serum ferritin was not raised in 15 of the 17 non‐splenectomized individuals. This group included one female who had been venesected for iron overload and further investigations have shown that both the genes for HS and haemochromatosis are present in her family.

These results demonstrate that iron stores are usually normal in HS, and that prevention of iron overload alone is not an indication for splenectomy. The rare report of a patient with HS and severe iron overload may perhaps be explained by the fact that the gene frequency for haemochromatosis is common in the population.
Viscosity and Iron Deficiency in Treated Polycythaemia
Authors: Pearson T. C. , Grimes A. J. , Slater N. G. P. , Wetherley‐Mein G.
Published: March 2008

Summary. An in vitro study of the effect of red‐cell changes due to iron deficiency on whole‐blood viscosity has been made on samples from patients with polycythaemia treated by venesection. When the PCV of the samples was adjusted to a standard value of 0.45, whole‐blood viscosity was unrelated to decreasing MCH. In contrast, when samples were adjusted to a standard Hb concentration of 14 g/dl, whole‐blood viscosity rose exponentially with decreasing MCH. This increase in whole‐blood viscosity was shown to be a function of the increasing PCV that accompanies the falling MCH at this standard Hb value. It is suggested that in polycythaemia not due to hypoxia, hyperviscosity can be satisfactorily corrected by venesection controlled by monitoring the PCV alone. Other considerations may apply in hypoxic polycythaemia and these are discussed.