bluuebunny/crossref_metadata_2025 huggingface dataset:
A New Method for the Simultaneous Quantitation of Platelet‐bound Immunoglobuin (IgG) and Complement (C3) Employing an Enzyme‐linked Immunosorbent Assay (ELISA) Procedure
Authors: Nel J. D. , Stevens K.
Published: March 2008

Summary.

An ELISA technique for the quantitation of platelet‐bound IgG and C3 is described. This is an antiglobulin consumption assay using commercial horseradish peroxidase conjugated anti‐IgG and anti‐C3 antisera. The greater the amount of antiglobulin consumed by the reaction with platelets the less is available to bind to an immune adsorbent in the form of human serum‐coated polystyrene balls. This test can be calibrated by adding known quantities of IgG or C3 to the specific enzymelinked antibody and the unbound fraction of antiserum is quantitated by allowing it to react with a chromogenic substrate for the enzyme and measuring the intensity spectrophotometrically, thus establishing an inverse relationship with the amount of antigen. Platelets from normal donors and those with non‐immunological throm‐bocytopenia gave values of 1·3–15·5 ng IgG/106 platelets and 0·22–0·96 ng C3/106 platelets which are in accordance with the normal ratio of these proteins in normal serum. Fifteen patients in whom immune destruction of platelets was suspected had excess platelet‐bound IgG ranging from 30 to 450 ng IgG/106 platelets. In none of these patients could excess platelet‐bound C3 be demonstrated. Compared to the antiglobulin consumption test we have found this test to be superior both technically and in terms of sensitivity and reproducibility.
Rapid Separation of Cytosol and Particle Fraction of Human Platelets by Digitonin‐induced Cell Damage
Authors: Akkerman J. W. N. , Ebberink R. H. M. , Lips J. P. M. , Christiaens G. C. M. L.
Published: March 2008

Summary.

Current cell disruption and fractionation techniques are time consuming and unsuitable for metabolic studies. We have developed a rapid method for platelets in which separation of cytosol and particle fraction is obtained within 50 s. Isolated platelet suspensions were incubated with low concentrations of digitonin followed by separation of soluble and particle fraction by centrifugation through a phthalate layer. Cell disruption was 90·1±4–2% (mean ± SD, n= 18; lactate dehyd‐rogenase leakage). Contamination of granules: acid hydrolase vesicles 16·2±3·6% (n=18, β‐N‐acetylglucosaminidase), dense granules 7–9% (n=3, 14C‐serotonin), mitochondrial matrix 0·6±0·1% (n=18, glutamate dehydrogenase). Low concentrations of digitonin did not affect sialic acid content, nucleoside diphosphate kinase and phosphodiesterase activity in isolated membranes. The method showed that most enzymes of glycolysis and hexose monophosphate shunt were localized in the cytosol except for hexokinase (96% particle bound), phosphoglucose isomerase (10% bound) and glutathion reductase (26% bound). About half the total ATP + ADP and most glycolytic intermediates were found partly particle bound, especially fructose 1,6‐diphosphate (40% bound). The data suggest that in platelets glycolysis occurs in different cell compartments.
Immune Complexes in Aplastic Anaemia
Authors: Cappio F. Caligaris , Novarino A. , Camussi G. , Gavosto F.
Published: March 2008

Summary. Immune complexes (IC) have been detected in nine out of 15 patients presenting with idiopathic aplastic anaemia using the polymorphonuclear neutrophil immunohistochemical technique. Immunosuppressive treatment undertaken in one patient produced a gradual recovery, the bone marrow repopulation being paralleled by the disappearance of IC. The significance of IC in aplastic anaemia and the relationship with possible pathogenesis and therapy of the disease are discussed.
Stability of International Reference Thromboplastins
Authors: van den Besselaar A. M. H. P. , van Halem‐Visser L. P. , Loeliger E. A. , Hermans J.
Published: March 2008

Summary. SUMMARY. Long‐term stability of the WHO International Reference Preparation (IRP) of thromboplastin 67/40 and of two secondary preparations, WHO IRP 68/434 and the Dutch RP 73/528 was assessed by two criteria on the basis of data obtained in the period 1969‐79. The clotting times of normal plasmas determined with these preparations over this 10 year period did not show changes which were of practical importance. Moreover, the slopes of the lines relating clotting‐time ratios of 67/40 to ratios obtained with 68/434 or 73/528 did not give any firm reasons to assume a systematic change of these slopes with time. These lines were determined by linear regression analysis of individual patient ratios. Corrections were applied to the original data for modifications of the assay technique made during the past 10 years.
The Decision to Perform Staging Laparotomy in Symptomatic Hodgkin's Disease
Authors: Rutherford Cynthia J. , Desforges Jane F. , Davies Byron , Barnett Arnold I.
Published: March 2008

Summary

. This study defines patients with symptomatic Hodgkin's disease for whom risks of staging laparotomy (LAP) outweigh benefits conferred by accurate knowledge of stage. From a database of more than 900 pathologically‐staged patients, probabilities of pathological stage are calculated for combinations of basic findings and lymphangiogram results. Decision‐making thresholds are defined at which results of treatment after LAP, taking operative mortality into account, are equivalent to immediate treatment appropriate to clinical stage. These thresholds are substantially altered by varying LAP mortality estimates, by assigning a false negative rate to LAP, and by considering uncertainty in treatment results. Fifty‐four combinations of findings are described for which immediate therapy with MOPP is justified; total nodal irradiation (TNI) is never indicated in B patients without prior LAP staging. Analysing 94 B patients who had LAP showed an appreciable number might have been spared this, particularly when uncertainty in treatment results is considered. With 0.03 (= 3%) uncertainty in treatment results, and 1% LAP mortality, LAP was not indicated in one in seven patients; nearly one third of patients should have immediate treatment with 3% LAP mortality. Threshold analysis can define those patients for whom risks of LAP outweigh benefits.
Impaired Neutrophil Function and Myeloperoxidase Deficiency in Pregnancy
Authors: El‐Maallem Hilal , Fletcher John
Published: March 2008

Summary

The ability of neutrophils to phagocytose and kill Candida guilliermondii was investigated in 28 pregnant women and compared with 29 healthy controls. Phagocytosis was normal, but the ability to kill and digest Candida was reduced to 65% of normal. Quantitative myeloperoxidase measurement revealed a considerable reduction of this enzyme (55% of normal) while neutrophil alkaline phosphatase levels were nearly twice as high as in normals. It is suggested that both lack of myeloperoxidase and impaired candidacidal ability result from the discharge of the neutrophil's granules during previous phagocytosis of antibody—antigen complexes in the circulation.
Abolished Production of Interferon by Leucocytcs of Patients with the Acquired Cytogcnetic Abnormalities 5q — or — 5 in Secondary and de‐Novo Acute Non‐lymphocytic Leukaemia
Authors: Pedersen‐Bjergaard J. , Haahr S. , Philip P. , Thomsen M. , Jensen G. , Ersboll J. , Nissen N. I.
Published: March 2008

Summary.

Interferon production by leucocytes on stimulation with inactivated antigens from herpes simplex virus, varicella zoster virus and cytomegalovirus was determined in 19 patients with preleukaemia or acute non‐lymphocytic leukaemia, in 15 following treatment for other tumours. Production of interferon was abolished in 10 patients and prcscrved in nine cases after stimulation with herpes simplex antigen. Cytogenetic studies of bone marrow demonstrated an abnormal karyotype in 13/15 patients with secondary preleukaemia or leukaemia. Characteristics were (a) hypodiploid cell lines demonstrated in eight cases, (b) a B‐chromosome defect found in five cases and verified as 5q— in four, and (c) defects in C‐group chromosomes in 10, verified as monosomy 7 in nine. All four patients with de‐novo preleukaemia or leukaemia had abnormal cytogenetic findings, two B‐chromosome abnormalities verified as monosomy 5. A relationship between abnormalities in chromosome no. 5 and abolished production of interferon was demonstrated, as only one of seven patients with 5q—, —5 or — B produced interferon by the leucocytes, compared with eight of a total of 12 patients with other cytogenetic defects or a normal karyotype (P=0.04). The results are remarkable, as chromosome no. 5 has recently been discovered to contain a gene for interferon production. Furthermore abolished production of interferon by leucocytes seemed correlated to previous irradiation and to disease‐related fever.
ADP Induced Surface Charge Changes of Adult and Newborn Platelets
Authors: Kosztoiányi G. , Jobst K. , Kellermayer M. , Ludány A.
Published: March 2008

Summary.

In experiments reported here electrophoretic mobility of washed platelets and platelets suspended in diluted plasma obtained from adults and newborns was practically the same. No significant difference was observed in the pH—mobility relationship of the two types of platelets. These comparative studies indicate that the actual charge density, i.e. the number and sign of the charged groups at the newborn and adult platelet surfaces are essentially identical.

However, a significant difference was found between the two platelet populations in the mobility changes induced by ADP. On the basis of cross over experiments between the platelets and plasma of adults and newborns it seems likely that the different behaviour of neonatal platelets arises from a difference between adult and newborn plasma. Preliminary results indicate that this may be due to the presence in adult plasma of a component with molecular weight about 10 000.
Red Cell Mass, Plasma Volume and Blood Volume before and after Venesection in Relative Polycythaemia
Authors: Humphrey P. R. D. , Michael J. , Pearson T. C.
Published: March 2008

Summary.

Seven male patients with relative polycythaemia have had red cell mass, plasma volume and blood volume measurements made before and after venesection. The mean PCV prior to venesection was 0.522. Using a small volume (250 ml) serial venesection technique, in each the PCV was reduced to or below 0.45, the overall mean value being 0.431. Venesection resulted in a rise in plasma volume with no significant reduction in blood volume in six of the patients. Thus hypovolaemia does not occur in the majority of the patients. One patient did not increase his plasma volume on packed cell volume reduction. His blood volume fell by 15%. None of the patients developed any symptoms of hypovolaemia. To maintain the PCV at values less than 0.45, venesection was required on average once every 2 months. It is suggested that venesection of small volumes is an acceptable long‐term method of lowering the PCV in patients with relative polycythaemia.
Marrow Cells from Patients with Untreated Pernicious Anaemia Cannot Use Tetrahydrofolate Normally
Authors: Deacon Rosemary , Chanarin I. , Perry Janet , Lumb Michael
Published: March 2008

Summary.

Folate analogues were added to human bone marrow cells to determine their effect on deoxyuridine utilization in the deoxyuridine suppression test. Forniyltetrahydrofolates fully corrected the impairment of dU utilization in pernicious anaemia marrows but tetrahydrofolate was relatively ineffective. All these analogues were effective in megaloblastic marrows from folate deficient patients.

Formyltetrahydrofolates also enhanced dU utilization by normal human marrows whereas methyltetrahydrofolate reduced its use.

In terms of the methylfolate trap hypothesis, the expectation that cobalamindeficient marrows would be able to use tetrahydrofolate normally was not realized.
Isoelectric Focusing of Hexokinase and Glucose‐6‐Phosphate Dehydrogenase Isoenzymes in Erythrocytes of Newborn Infants and Adults
Authors: Gahr M.
Published: March 2008

Summary.

Isoenzymes of glucose‐6‐phosphate dehydrogenase and hexokinase from red cells of newborn infants were analysed by isoelectric focusing in polyacrylamide gel after partial purification. The pattern of isoenzyme distribution was compared with that of erythrocytes from adults. Glucose‐6‐phosphate dehydrogenase isoenzyme distribution was identical between erythrocytes from newborn infants and adults. Erythrocytic hexokinase isoenzymes patterns were different between newborn infants and adults. Erythrocytes from adults contain a hexokinase isoenzyme which has the same isoelectric point as rat liver hexokinase isoenzyme I (pH 6.01). This isoenzyme is lacking in red cells from newborn infants. Isoenzymes with the isoelectric properties of rat liver isoenzyme II (pH 5.48) were not detectable in red cells from adults, nor from newborn infants. The occurrence of an isoenzyme III in red cells remained unclear, because only a faint staining was observed in the corresponding region of erythrocytic gels. Preliminary results revealed a fetal type of hexokinase isoenzyme distribution in infants with an age of 10 months. This indicates that regulation of the synthesis of Hb F and of fetal hexokinase isoenzynies is not correlated.
Sinus Histiocytosis with Massive Lymphadenopathy: Virological, Immunological and Morphological Studies
Authors: Karpas Abraham , Worman Colin , Arno Jeanne , Nagington Jack
Published: March 2008

Summary The histiocytes from a patient with sinus histiocytosis were tested for the presence of Epstein‐Barr viral nuclear antigen (EBNA) before and after in vitro growth. On both occasions the histiocytes as well as the lymphocytes were EBNA negative. Likewise the profile of EBV viral capsid antibody (VCA) suggests that this virus is unlikely to be directly involved in the development of the disease. The maintenance of an unusually high level of antibodies to EBV and measles, both viruses which produce persistent infections, indicates that the humoral immune response is active. That a normal humoral response occurred in response to mumps virus during the same period suggests that a deficiency in the cell mediated immune response to the persistent viruses might be the reason for the abnormally high antibody levels. The fresh as well as the cultured histiocytes had only Fc receptors. The affected lymph node lymphocytes developed unusual large inclusion bodies following in vitro culture.
Peripheral Blood Cells from Patients with Aplastic Anaemia in Partial Remission Suppress Growth of their Own Bone Marrow Precursors in Culture
Authors: Nissen Catherine , Cornu Pierre , Gratwohl Alois , Speck Bruno
Published: March 2008

Summary In 12 patients with severe aplastic anaemia who had achieved self‐sustaining autologous bone marrow function after treatment with antilymphocyte globulin, or with cyclophosphamide given for attempted bone marrow transplantation, colony formation by all haemopoietic precursors remained far below normal. Precursors from peripheral blood, erythroid precursors in particular, failed to form a normal number of colonies. This paucity of colony formation does not reflect a true lack of precursor cells but is due to circulating cells which impair maturation. Addition of low density peripheral blood cells to autologous bone marrow cultures diminished colony formation by granulocyte‐macrophage precursors (GM‐CFC) and abolished ‘burst’ formation by BFU‐E. Strong auto‐inhibition preceded relapse in four of eight patients. The phenomenon was not observed in five normals, in five aplastic anaemia patients with stable haemopoietic grafts and in three polytransfused control patients.

The T‐cell poor subpopulation of peripheral blood cells, containing mainly B‐cells and macrophages, was especially inhibitory. Accordingly, removal of plastic adherent cells from bone marrow cell suspensions improved plating efficiency in aplastic anaemia patients, but not in normals. Isolated E‐rosette positive cells had no negative effect.

Inhibition could only be demonstrated in the autologous situation. Colony formation by normal allogeneic peripheral blood precursors was not impaired by patient cells. The phenomenon is likely to reflect residual disease activity which is compensated in vivo but can be demonstrated in vitro. It may be of help in early recognition of patients who are at risk of relapse after autologous bone marrow reconstitution.
Biliary Iron Excretion in Rats following Pyridoxal Isonicotinoyl Hydrazone
Authors: Cikrt M. , Poňka P. , Nečas E. , Neuwirt J.
Published: March 2008

Summary Biliary excretion of iron after administration of pyridoxal isonicotinoyl hydrazone (PIH), a recently identified effective iron‐chelating agent, was investigated in rats. PIH administered both intraperitoneally and orally was shown to increase significantly 59Fe excretion into bile of rats which had previously been injected with 59Fe‐transferrin to label hepatic parenchymal cells. 59Fe‐PIH appears in bile as early as 15 min after chelator administration and the peak of 59Fe‐radioactivity in bile is seen 1–5 h following intraperitoneal PIH injection. PIH, administered intraperitoneally, 125–250 mg/kg, increased 24 h biliary radioiron excretion about 35 times and in addition increased urinary and faecal iron excretion. When PIH was given immediately before 59Fe‐transferrin, 24 h cumulative biliary 59Fe excretion was even higher. PIH was also demonstrated to increase biliary excretion of radioiron released from 59Fe‐haemoglobin catabolysed in reticuloendothelial cells. The effect of PIH was confirmed by estimation of biliary iron concentration using the method of atomic absorption spectrophotometry. Repeated PIH administration to rats decreased 59Fe radioactivity in liver and kidney and increased urinary and faecal iron excretion.
In Vitro Inhibition of Granulopoiesis by 4‐Amino‐4′‐Hydroxylaminodiphenyl Sulfone
Authors: Weetman Robert M. , Boxer Laurence A. , Brown Mary P. , Mantich Nancy M. , Baehner Robert L.
Published: March 2008

Summary The antimalarial drug, 4,4′‐diaminophenylsulfone (dapsone) has been associated clinically with the production of agranulocytosis. A metabolite of dapsone, 4′‐amino‐4′‐hydroxylaminodiphenyl sulfone (DDS‐NOH) generates superoxide anion and hydrogen peroxide in vitro. To link oxidant injury of committed myeloid stem cells to agranulocytosis, the effects of DDS‐NOH on altering human bone marrow CFUc proliferation and in vitro reduced glutathione levels were examined in the presence and absence of superoxide dismutase and catalase. Bone marrow cells were preincubated for 60 min with either 0·01, 0·1 or 1·0 mm DDS‐NOH, washed and then 2×105 cells/plate were cultured for 10–14 d. Leucocyte colony forming unit cell (CFUc) proliferation was markedly suppressed by 0·1 mm and 1·0 mm DDS‐NOH. With 0·1 mm DDS‐NOH, addition of superoxide dismutase to the preincubation mixture further decreased colony numbers, while addition of catalase partially restored colony formation indicating involvement of hydrogen peroxide. After exposure to 0·1 mm DDS‐NOH, trypan blue exclusion by nucleated marrow cells was 98%. At the same DDS‐NOH concentration, addition of lactoperoxidase, sodium iodide and superoxide dismutase to the preincubation mixture resulted in a further decrease in CFUc proliferation. After incubation with 1·0 or 2·0 mm DDS‐NOH, bone marrow reduced glutathione levels were markedly decreased from 325 nmol/108 cells to 116·3 nmol/108 cells. Bone marrow reduced glutathione levels in guinea‐pig cells also fell after exposure to 1·0 and 2·0 mm DDS‐NOH. In turn catalase but no superoxide dismutase partially prevented the fall. Thus, these studies demonstrate that oxidant‐induced suppression of human marrow CFUc proliferation in vitro can be mediated by a metabolite of dapsone, DDS‐NOH.
Active Immunotherapy for the Treatment of Acute Myelogenous Leukaemia: Report of Two Controlled Trials
Authors: Whittaker J. A. , Bailey‐Wood R. , Hutchins S.
Published: March 2008

Summary Over 61/2 years, 182 patients with acute myelogenous leukaemia were treated with one of two combinations of chemotherapy containing cytosine arabinoside and an anthracycline (daunorubicin or doxorubicin). Eighty‐one patients achieved remission and 79 of them were entered into one of two trials of active immunotherapy. The first trial compared maintenance chemotherapy and i.v. BCG immunotherapy with chemotherapy alone. The results have shown that the group given i.v. BCG survived for a significantly longer time (P=0·035) than the group treated only with chemotherapy. The i.v. BCG treated group also had a significantly longer survival (P=0·042) after their first relapse and a higher incidence of subsequent remissions. However, there was no difference in the length of first remissions for the two groups.

The second trial compared two different types of active immunotherapy. The results show no significant difference in remission duration or survival after first relapse for 34 patients randomly allocated to treatment with i.v. BCG or irradiated leukaemic blast cells. Sixteen patients relapsed and subsequently 10 patients entered a second remission after reinduction chemotherapy. These patients were distributed evenly between the two immunotherapy groups and this high rate of second remissions is similar to that for the immunotherapy group in the first Cardiff Trial. In the two trials, 21 (62%) of 34 patients receiving immunotherapy entered a second remission after reinduction chemotherapy and six patients achieved third remissions.
Genetic and Biochemical Heterogeneity of β‐Thalassaemia in Naples
Authors: Pepe G. , Lupi L. , Luzzatto L.
Published: March 2008

Summary We have investigated 32 children with Cooley's anaemia from Naples, Italy. Criteria for inclusion in the study were: (a) typical clinical and haematological findings; (b) absolute transfusion requirement; and (c) elevated Hb A2 in both parents. From biosynthetic studies we have established that five children (including two sets of sibs) had β° thalassaemia, while the rest had β+ thalassaemia. Thus, the frequency of β+ thalassaemia among unrelated patients was about 90%. The distribution of β/α ratios among β+ patients ranged from 0·01 to 0·16 and it was bimodal, consistent with some of them having a β°/β+ genotype and others a β+/β+ genotype. The distribution of β/α ratios of the patients' parents (obligate heterozygotes) ranged from 0·24 to 0·73, and it was plurimodal, consistent with the coexistence in this population of multiple β thalassaemia alleles, of which one must be β° and at least one is β+. A systematic analysis of 20 families indicates that the β/α ratio is to some extent quantitatively inherited, and its suggests non‐randomness in the assortment of β thalassaemia alleles that can give rise to a Cooley's phenotype.
Thrombopoiesis and Megakaryocyte Colony Stimulating Factor in the Urine of Patients with Aplastic Anaemia
Authors: Enomoto Katsuto , Kawakita Makoto , Kishimoto Susumu , Katayama Noritaka , Miyake Takaji
Published: March 2008

Summary The urinary extracts from patients with aplastic anaemia and from healthy donors were investigated in vivo and in vitro for their ability to stimulate megakaryopoiesis and platelet production. There was a significantly higher concentration of thrombopoiesis stimulating factor (TSF) and megakaryocyte colony stimulating factor (MEG‐CSF) in the urine from patients with aplastic anaemia than in that from healthy donors. Neuraminidase treatment did not affect the thrombopoietic activity of TSF, whereas coexisting erythropoictin (EPO) in the extract lost its activity in vivo. These findings suggest that TSF and/or MEG‐CSF seems to be different from EPO and that the urine from aplastic anaemia patients would be a good source of TSF and MEG‐CSF for purification and characterization.
Seven Pyruvate Kinase Variants Characterized by the ICSH Recommended Methods
Authors: Miwa S. , Fujii H. , Takegawa S. , Nakatsuji T. , Yamato K. , Ishida Y. , Ninomiya N.
Published: March 2008

Summary Seven new red‐cell pyruvate kinase (PK) variants were characterized by the methods recently recommended by the International Committee for Standardization in Haematology. The cases were all true homozygote as evidenced by consanguineous marriages of the parents; all are Japanese. These variants were designated as PK Tokyo, PK Nagasaki, PK Sapporo, PK Maebashi, PK Itabashi, PK Fukushima and PK Aizu, respectively. Low substrate affinity (high K0.5S for phosphoenolpyruvate) and thermal instability appear to play major roles in causing defective enzyme function, resulting in chronic haemolytic anaemia. Product inhibition of PK by ATP may also play an additional role in causing haemolysis in more than half the cases.
Intensive Plasma Exchange in the Management of Severe Rh Disease
Authors: Angela E. , Robinson E. , Tovey L. A. D.
Published: March 2008

Summary Fourteen high‐risk cases of Rh alloimmunized women were treated by intensive plasma exchange on the cell separator throughout their pregnancies. The mean volume of plasma exchanged per week was 3·21 with a total volume of 10—123 l. The replacement fluid was mainly plasma protein fraction (PPF) supplemented with fresh frozen plasma (FFP). The mean duration of treatment was 13 1/2 weeks, ranging from 2 to 22 weeks, commencing at 12—30 weeks gestation. The complications encountered and the selection of the optimal time for delivery are discussed. The expected stillbirth rate in this series as determined by their past obstetric histories and anti‐D levels was 62%. Intrauterine transfusion was given to only two of the infants and both were later stillborn. It was possible to reduce and maintain a lower level of anti‐D in the serum of most of the patients and a successful outcome was achieved in nine of the 12 cases included for analysis (75%).

Plasma exchange commenced early in pregnancy is recommended as a non‐hazardous form of treatment in the management of severe Rh haemolytic disease. However, if the mean level of anti‐D cannot be maintained at less than 35 iu/ml then the outcome is more likely to be fatal. Amniocentesis should be delayed where possible until 28 weeks gestation and intrauterine transfusion reserved for those cases where the anti‐D level becomes uncontrollably high and fetal death is imminent.
Hyperviscosity and Thrombotic Changes in Idiopathic and Secondary Raynaud's Syndrome
Authors: Blunt R. J. , George A. J. , Hurlow R. A. , Strachan C. J. L. , Stuart J.
Published: March 2008

Summary Whole‐blood viscosity (measured at 128, 23 and 0·2 s‐1) was significantly increased in 29 females with Raynaud's syndrome, 13 males with vibration‐induced Raynaud's syndrome, and 18 males with calf claudication secondary to atherosclerosis, as compared with 50 healthy controls matched for sex, age and smoking habit. Viscosity was higher at low temperature (27 and 22°C) in all three types of vascular disease and, despite symptoms of cold hypersensitivity, patients with Raynaud's syndrome did not show selective hyperviscosity at low temperature.

Patients with vascular disease, irrespective of aetiology, also showed an increase in the acute‐phase reactants haptoglobin, fibrinogen, and factor VIII antigen, together with reduced fibrinolytic activity and minor activation of platelets. These alterations in viscosity and haemostatic factors in vascular disease are probably related to the degree, rather than the aetiology, of endothelial damage and their cumulative effect may contribute to local stasis and thrombosis, particularly in cold extremities.
The Role of Intensive Plasma Exchange in the Prevention and Management of Haemorrhage in Patients with Inhibitors to Factor VIII
Authors: Slocombe G. W. , Newland A. C. , Colvin M. P. , Colvin B. T.
Published: March 2008

Summary. A strategy for the prevention and management of haemorrhage in patients with inhibitors to factor VIII by intensive plasma exchange and human factor VIII infusion is described. The advantages and disadvantages of this approach are discussed in the light of the authors’ experience with the technique and the alternative methods of treatment described in the literature.
Constitutional Thrombocytopathy with Subnormal Response to Thromboxane A2
Authors: Samama M. , Lecrubier C. , Conard J. , Hotchen M. , Breton‐Gorius J. , Vargaftig B. , Chignard M. , Lagarde M. , Dechavanne M.
Published: March 2008

Summary. A new type of congenital platelet dysfunction was found in a young woman presenting a life‐long bleeding disorder. The known types of thrombopathia and von Willebrand's disease were excluded by appropriate investigations. The platelets were morphologically normal, underwent normal shape change and contraction and synthesized thromboxane A2 (TXA2) normally. The release reaction was abnormal and the aggregation response to ADP, adrenalin, collagen, thrombin, sodium arachidonate and vasopressin was depressed due to decreased sensitivity of the platelets to prostaglandin endoperoxides and TXA2. Platelet cAMP content was increased.
Treatment of Refractory Adult Acute Lymphocytic Leukaemia and Acute Undifferentiated Leukaemia with an Anthracycline Antibiotic and Cytosine Arabinoside
Authors: Early Amy Papalia , Preisler H. D. , Gottlieb Arlin J. , Lanchant Neil A.
Published: March 2008

Summary. Ten patients with acute lymphocytic leukaemia (ALL) and four patients with acute undifferentiated leukaemia (AUL) in relapse or refractory to conventional therapy were treated with remission induction therapy consisting of an anthracycline antibiotic and cytosine arabinoside. Twelve patients had previously demonstrated resistance to vincristine‐prednisone and nine patients had prior anthracycline therapy. Nine patients achieved complete remission after one course of therapy with a median time to remission of 30 d. Of five nonresponders, three died of sepsis with marrow hypocellularity and no evidence of residual leukaemia. Only two patients had unequivocal evidence for resistance to an anthracycline—cytosine arabinoside regimen. Myelosuppression and infection were the most significant complications of therapy. The data presented indicate that marrow ablative chemotherapy with an anthracycline antibiotic and cytosine arabinoside is an effective regimen for remission induction in adults with ALL and AUL refractory to vincristine—prednisone. The use of these agents in remission consolidation therapy may offer the possibility of providing a reduction in residual resistant cells that are present after successful remission induction therapy with conventional agents.
Erythropoietin Measurements in the Differential Diagnosis of Polycythaemia
Authors: Napier J. A. F. , Janowska‐Wieczorek A.
Published: March 2008

Summary. An in vitro bioassay has been used to measure erythropoietin levels in extracts of serum from 74 patients with polycythaemia. Patients with polycythaemia vera in relapse had subnormal or undetectable levels which did not increase after challenge by venesection. In polycythaemia secondary to hypoxaemia serum erythropoietin concentrations were generally elevated with a further elevation occurring after venesection. Where polycythaemia was secondary to renal polycystic disease or tumours increased levels of erythropoietin could not be consistently demonstrated. In these patients the serum erythropoietin was unaffected by venesection. In other patients in whom the cause for polycythaemia was unknown examples of each of the above patterns were seen. Erythropoietin investigations in such patients may help in the elucidation of the underlying pathology.
Autologous Neutrophils Inhibit Production of Colony Stimulating Activity by Normal Human Lymphocytes
Authors: Lutynski A. , Th'ng K. H. , Goldman J. M.
Published: March 2008

Summary. Experiments were designed to study the way in which normal human polymorphs (PMNs) inhibit the production or release of colony stimulating activity (CSA) by normal lymphocytes incubated alone or in conjunction with normal monocytes. PMNs were first incubated with lymphocytes for varous periods at various concentrations. The PMNs were then removed and the ‘conditioned’ lymphocytes were used alone or after addition to adherent mononuclear cells (monocytes) for the production of conditioned medium. The samples of conditioned medium were then assayed for CSA in a standard system for culturing granulocyte/macrophage progenitor cells (CFU‐C) in agar. We found that the capacity of PMNs to inhibit CSA production by lymphocytes or by lymphocytes plus monocytes was directly proportional to the number of PMNs originally incubated and maximal at relatively short incubation times (i.e. 2–4 h). Such inhibition could be counteracted by the introduction of known stimulators of CSA production by monocytes, e.g. phytohaemagglutinin or bacterial toxin.

We conclude that normal lymphocytes ‘conditioned’ or ‘programmed’ by contact with PMNs may themselves have a reduced capacity to produce CSA and may also act to reduce CSA production by monocytes. Such programmed lymphocytes could thus be a component of the mechanism by which PMNs exert a physiological inhibition on CSA‐dependent granulopoiesis in vivo.
Differentiation of Cell Surface Receptors on Normal Human Bone Marrow Myeloid Precursors
Authors: Barrett Sondra G. , Hansen Keith S. , Bainton Dorothy F.
Published: March 2008

Summary. Receptors for IgG and complement are present on the plasma membrane of functionally mature blood neutrophils. Neutrophilic precursors lack these membrane receptors, which may account for their diminished phagocytic capability. In this study, myeloid precursors from human bone marrow were examined for the presence and activity of membrane receptors for IgG and activated complement. Morphologic identification by light microscopy of neutrophilic cells was made by rosetting techniques and chloroacetate esterase cytochemical staining and at the ultrastructural level by peroxidase reactivity.

This study showed that the Fc receptor was the only membrane marker on the myeloblast and that the number of cells with this receptor increased with maturation. Functional investigation of the Fc receptor indicated that its presence on early myeloid cells was insufficient to induce IgG‐mediated ingestion. Phagocytosis via the Fc receptor was not acquired until the metamyelocyte stage. Complement receptors, C3d and C3b, appeared later than the Fc receptor. The C3d receptor, detected first on the promyelocyte, remained relatively constant throughout maturation, whereas the C3b receptor acquired at the myelocyte stage increased with maturation.

These data provide evidence that distinct membrane changes occur during normal myeloid cell maturation and that the presence of these membrane receptors is not sufficient for Fc‐mediated ingestion to occur.
The Development of Haematological Changes in Homozygous Sickle Cell Disease: a Cohort Study from Birth to 6 Years
Authors: Serjeant Graham R. , Grandison Yvonne , Lowrie Yvonne , Mason Karlene , Phillips Jennifer , Serjeant Beryl E. , Vaidya Shuba
Published: March 2008

Summary.

A cohort study of sickle cell disease from birth has allowed observations on the disease without the symptomatic selection inherent in previous series. The development of haematological indices from birth to 6 years in male and female infants with homozygous sickle cell (SS) disease is presented and compared with values in age and sex matched controls with a normal haemoglobin (AA) genotype previously presented elsewhere. In SS disease total haemoglobin levels fell rapidly from birth to a plateau at 3‐6 months before falling again to 15 months after which no age related change occurred. Mean cell haemoglobin concentration fell from birth to lowest values at 15‐18 months before increasing to reach the level present at birth by the age of 5 years. Red cell counts fell rapidly after birth to a plateau at 2 months, increased slightly to 6 months and then fell steadily throughout the remaining period of the study. The mean cell volume and mean cell haemoglobin also fell rapidly after birth reaching the lowest values by 6 months and then increased progressively. Female patients showed significantly higher MCV from 4 to 8 months and significantly higher haemoglobin levels from 15 months to 41/2 years. Compared to AA controls, SS patients manifested significantly lower levels of haemoglobin from 2 weeks, and red cell counts from 1 month, and significantly higher levels of MCHC from 4 months to 3 years, MCV from 8 months to 5 years, and serum iron levels from 1 to 4 years. Children with SS disease were partially protected from iron deficiency in early childhood, perhaps by increased intestinal absorption of iron, and the associated increase in intracellular haemoglobin concentration might be disadvantageous during this high risk period.
Deficiency of Plasma Prostacyclin or PGI2 Regenerating Ability in Sickle Cell Anaemia
Authors: Stuart Marie J. , Sills Richard H.
Published: March 2008

Summary.

We have demonstrated that patients with sickle cell disease > 2 years of age have a marked decrease in their plasma PGI2 or prostacyclin regenerating ability (0.05 ± 0.08 ng/mg of vascular tissue) when compared to normal controls (0–44 ± 0.12). This abnormality was not present in other chronic haemolytic states, and was not related to concomitant hyposplenism. Normal activity was observed in patients with sickle cell disease following a period of prophylactic red cell transfusions (0.32 ± 0.13). Since vascular prostacyclin is antiaggregatory and antithrombotic, the deficiency in the prostacyclin regenerating ability of patients with sickle cell disease may play a pathogenic role in the development of the microvascular and thrombotic complications seen in this disorder.
Synthesis and Metabolism of Bence Jones Protein and Calculation of Tumour Burden in Patients with Bence Jones Myeloma
Authors: Durie Brian G. M. , Cole Perry W. , Chen H.‐S. George , Himmelstein Kenneth J. , Salmon Sydney E.
Published: March 2008

Summary. Synthesis and metabolism of Bence Jones protein was measured in seven patients with Bence Jones myeloma. Using the plasma and urine levels of 125I‐labelled Bence Jones protein, it was possible to calculate the fractional catabolic rate (FCR), fractional proteinuric rate (FPR), and fractional metabolic rate (FMR) for the individual patients. There was a high degree of correlation between decreased metabolism and reduced creatinine clearance (P < 0.001). Ideal regression equations (linear and nonlinear) relating FCR, FPR and FMR with fractional creatinine clearance (FCC) were calculated. Using these regression equations plus measured values for the synthetic rates of light chain for bone marrow plasma cells, the myeloma cell mass or tumour burden was measured in each patient. Correlations between these measured cell mass values and those derived from clinical staging were reasonably good. However, for the lambda (γ) Bence Jones myeloma patients, the measured cell mass values were significantly higher than predicted from clinical staging. Further studies will be necessary to validate this latter observation. It is hoped that this study will serve as a basis for a comprehensive schema allowing accurate cell mass measurement and staging of Bence Jones myeloma.
The Switch from Haemoglobin F to A: the Time Course of Qualitative and Quantitative Variations of Haemoglobins after Birth
Authors: Terrenato L. , Bertilaccio C. , Spinelli P. , Colombo B.
Published: March 2008

Summary. The time course of the transition from HbF to HbA production in the postnatal period has been studied. The description is based on the time‐dependent pattern of total haemoglobin and of HbF observed in an 8 months follow‐up of 25 premature newborns. The absolute amount of HbF decreases exponentially from birth to approximately 25 weeks (from 15.56 0.48 g/dl to 0.66 0.08 g/dl), at a weekly rate of approximately 16%. On the other hand, HbA shows a biphasic pattern: first it decreases slightly from birth to 6 weeks (from 4.09 0.32 g/dl to 2.63 0.33 g/dl); afterwards it increases exponentially at a weekly rate of 5% up to a plateau value, until HbF is almost completely replaced by HbA. Therefore the total haemoglobin decreases exponentially from birth to approximately 6 weeks (from 19.65 0.46 to 9.92 0.31); it remains approximately constant for the next 6 weeks, and thereafter it increases very slowly.

From these data, two distinct phases of postnatal erythropoiesis can be identified: the first (from birth to 6 weeks) is characterized by a decrease in the total amount of haemoglobin produced whereas in the second there is an alteration in the relative proportions of HbF and HbA being synthesized.
The Genetics and Molecular Basis of Alpha Thalassaemia in Association with Hb S in Jamaican Negroes
Authors: Higgs D. R. , Pressley L. , Serjeant G. R. , Clegg J. B. , Weatherall D. J.
Published: March 2008

Summary. We have studied seven Jamaican Negro families in whom the genes for α thalassaemia and the sickle cell mutation (βs) were independently segregated. Using a combination of techniques we identified two α thalassaemia phenotypes which resemble the severe (α thalassaemia 1) and mild (α thalassaemia 2) determinants previously described in Orientals. This study has enabled us to clearly correlate the phenotype of α thalassaemia with the genotype in this population. Furthermore, since in each family α thalassaemia was present in association with the gene for the sickle cell mutation we have determined the proportion of Hb S in the peripheral blood of individuals with the αα/αα, ‐α/αα and ‐α/‐α genotype who are also heterozygous for the βs mutation. Genetic analysis in these families shows that in each case subjects with the α thalassaemia 1 phenotype are homozygous for the α thalassaemia 2 defect (‐α/ ‐α). We have found no instances of the genotype ‐/αα in this population which may explain the rarity of the severe α thalassaemia syndromes in Jamaica. Restriction mapping data in the α thalassaemia 2 homozygotes from this population shows that the (‐α/) haplotype results from a deletion of one of the linked pair of α globin genes and that this has probably arisen by an unequal crossover between non‐homologous α genes.
Immunofluorescent and Biochemical Studies of Terminal Deoxynucleotidyl Transferase in Treated Acute Leukaemia
Authors: Bradstock K. F. , Janossy G. , Hoffbrand A.V. , Ganeshaguru K. , Llewellin P. , Prentice H. G. , Bollum F. J.
Published: March 2008

Summary. Indirect Immunofluorescence (IF) for terminal deoxynucleotidyl trans‐ferase (TdT) was used in conjunction with the biochemical assay of TdT enzymatic activity to study human leukaemias before and during therapy. In addition, non‐leukaemic marrows were analysed to compare the enzyme expression on normal cells. An excellent correlation was observed between the IF and biochemical methods when specimens contained greater than 5% TdT+ cells (by IF); below this level the biochemical assay was less reliable, while the sensitive IF test could detect isolated TdT+ cells among greater than 10 000 TdT negative cells. The IF method also had the advantage of allowing further immunological characterization of TdT+ cells, by simultaneous labelling of membrane antigens with appropriate antiscra. TdT+ cells expressing la‐like antigens (but lacking other antigens associated with B‐ and T‐ lymphoid differentiation) were frequently found in low numbers in remission marrows from acute lymphoblastic leukaemia (ALL) patients. However, similar cells were also observed in remission acute myeloid leukaemia, as well as in non‐leukaemic regenerating marrows, and marrow from normal donors. The presence of these normal TdT+ precursor cells therefore precluded the use of either IF or biochemical TdT tests for estimating the degree of residual disease or predicting early relapse in patients with non‐T, non‐B ALL. In contrast, the detection of TdT+ cells with T lymphoid antigens (HuTLA+) but lacking Ia antigens, in thymic (T‐cell) ‐ALL, but not in normal marrow, allowed the use of this combination of markers to detect minimal residual disease in T‐ALL.
Terminal Deoxynucleotidyl Transferase Expression in Acute Non‐lymphoid Leukaemia: an Analysis by Immunofluorescence
Authors: Bradstock K. F. , Hoffbrand A. V. , Ganeshaguru K. , Llewellin P. , Patterson K. , Wonke B. , Prentice A. G. , Bennett M. , Pizzolo G. , Bollum F. J. , Janossy G.
Published: March 2008

Summary. Indirect immunofluorescence for terminal transferase enzyme (TdT) was used to study the blasts of 64 patients with acute non‐lymphoid leukaemia (ANLL). In 32 patients no TdT positive cells were seen. In 19 cases a small subpopulation of cells expressing TdT was detected; these constituted up to 5% of total nucleated cells, and it was not clear whether these TdT positive cells were part of the leukaemic process or represented residual normal bone marrow lymphoid cells.

The remaining 13 patients had TdT positive cells accounting for 7–90% of the total. In two of these cases TdT was expressed on blasts with myeloid features, representing an aberrant expression of TdT by myeloid cells; in contrast, in three cases mixed populations of TdT positive lymphoid blasts and TdT negative myeloid blasts were observed. In the remaining cases it was not possible to determine whether the TdT positive cells had definite lymphoid or myeloid features. Cytogenetic analysis showed no evidence of the Philadelphia chromosome. Response to treatment was assessed in 11 of the 13 patients. Only one patient remitted with the initial choice of therapy (DAT); four failed to respond to initial regimes of vineristine and prednisone (V & P) while the other five patients did not respond to myelotoxic combinations (DAT). Only one patient subsequently entered complete remission on second line therapy (V & P). This group of patients with TdT+ ANLL had a particularly bad prognosis, and appeared to differ from cases of TdT positive acute undifferentiated leukaemia, which often respond to V & P therapy.
Tumour Promoters Induce Macrophage Differentiation in Human Myeloid Cells from Patients with Acute and Chronic Myelogenous Leukaemia
Authors: Fibach E. , Rachmilewitz E. A.
Published: March 2008

Summary. Twelve‐O‐tetradecanoyl phorbol‐13‐acetate (TPA) and other tumour promoter plant diterpenes transformed myeloid cells from peripheral blood, bone marrow and spleen from patients with chronic and acute myeloid leukaemia and acute myelomonocytic leukaemia into macrophage‐like cells. This transformation resulted in cessation of cell multiplication, adherence of the cells to the surface of the culture dish, and acquisition of phagocytic activity, Fc receptors and enzymatic content characteristic of the monocyte‐macrophage pathway of differentiation. The fact that tumour promoters induce differentiation in human myeloid leukaemia cells and not in similar cells from non‐leukaemic conditions suggest their possible application in diagnosis and chemotherapy.
Effects of Trimethoprim on Leukaemic Cells in Vitro
Authors: Rivard Georges E. , Momparler Louise F. , Momparler Richard L.
Published: March 2008

Summary. Trimethoprim‐Sulfamethoxazole (TMP‐SMZ) is a fixed combination of antibiotics which is widely used for prophylaxis and treatment of infections in patients undergoing cancer chemotherapy. TMP has been reported to inhibit growth of haemopoietic stem cells in vitro. If TMP—SMZ inhibits leukaemic cell growth, it could interfere with antileukaemic treatment, especially with S phase specific agents. TMP—SMZ at a concentration of 10 μg/ml (TMP) produced 50% inhibition of incorporation of H‐deoxyuridine in DNA of L1210 and human lymphoblastic leukaemia cell. TMP‐SMZ (1 g/ml TMP) produced 30% prolongation of doubling time of L1210 in vitro. Pure TMP (10 μg/ml) but not SMZ (50 μg/ml) produced the same effect as TMP‐SMZ. Cell inhibitory effects could be completely reversed by folinic acid. These findings suggest that TMP produces some degree of inhibition of dihydrofolic acid reductase in mammalian cells and can potentially influence the effects of chemotherapy on tumour and/or host cells.