bluuebunny/crossref_metadata_2025 huggingface dataset:
The fate of excess β‐globin chains within erythropoietic cells in α‐thalassaemia 2 trait, α‐thalassaemia 1 trait, haemoglobin H disease and haemoglobin Q‐H disease: an electron microscope study
Authors: Wickramasinghe S. N. , Hughes Madeleine , And S. Fucharoen , Wasi P.
Published: March 2008

Summary. Electron microscope studies have been performed on bone marrow cells from individuals with various α‐thalassaemia syndromes. Globin chain precipitates were rarely found in the erythropoietic cells of a subject with α‐thalassaemia 2 trait. By contrast, such precipitates were found in 0.6–1.3% of the erythroblast and marrow reticulocyte profiles in two cases of α‐thalassaemia 1 trait, 2.1–13.7% of profiles in five patients with haemoglobin H (HbH) disease and 6.2% of profiles in one patient with haemoglobin Q‐H (HbQ‐H) disease. In the patient with HbQ‐H disease, but not in the others, the nuclei of some erythroblasts displayed the ‘Swiss cheese’appearance which has been reported in some forms of congenital dyserythropoietic anaemia. It is proposed that in α‐thalassaemia 2 trait, where the degree of excess of β‐chains is slight, most of the excess chains are degraded by proteolysis. In α‐thalassaemia 1 trait, where there is a greater excess of β‐chains, the excess chains can undergo either proteolysis or precipitation, but there is little or no tetramerization to form soluble HbH. Finally, in HbH disease, where the degree of excess is considerable, the excess β‐chains may be subject to proteolysis, precipitation and tetramerization.
Chromosome analysis of isolated colony erythroblasts in chronic myelogenous leukaemia
Authors: Dainiak Nicholas , Liu Aileen , Dewey Maureen C. , Kulkarni Vivekanand
Published: March 2008

Summary. To isolate and karyotype the progeny of erythroid progenitors, we applied colony erythroblasts derived from plasma clot marrow cultures from two healthy adults and two patients with newly diagnosed Ph1+ chronic myelogenous leukaemia (CML) to discontinuous Stractan density gradients. Erythroid colony proliferation by patient cells was increased relative to that of normal donor cells (P<0.01). ‘Endogenous’colonies appeared in patient but not in normal donor marrow cultures. Greater than 95% of nucleated cells equilibrating at 1.071 were basophilic proerythroblasts. While analysis of chromosome spreads of normal donor cells in this fraction showed normal karyotypes, cells from patient marrow cultures were Ph1+, whether cultured in the presence or absence of added erythropoietin. These findings suggest that chromosome abnormalities of erythroid progenitors may be expressed by their progeny in tissue culture, and that Stractan may be a useful supporting medium for separating colony erythroblasts for chromosome analysis.
Ph1‐positive and Ph1‐negative abnormal cell lines in a child with lymphoblastic lymphoma
Authors: Shikano Takaaki , Kaneko Yasuhiko , Ishikawa Yorikazu , Niikawa Norio , Tonooka Tatsuto , Takeda Takeo , Kikuchi Masahiro
Published: March 2008

Summary. A 7‐year‐old Japanese boy with Ph1‐positive‐lymphoblastic lymphoma is described. The diagnosis was based on biopsied tonsils which were enlarged at the time of admission. On the eighth day after admission an enlarged mediastimal mass was detected on a chest X‐ray film. The lymphoblasts which appeared in the peripheral blood and bone marrow proved to be T‐cells. Chromosome studies on the bone marrow cells revealed two abnormal cell lines; one had a 7; 11 translocation and the other a 7;11 translocation and a 9;22 translocation, forming the Ph1‐chromosome. The latter line with the Ph1 chromosome was considered to have been derived from the former line without the Ph1. Our findings show that the Prochromosome may be a secondary change in the course of karyotypic evolution.
Megakaryoblast precursors in rodent bone marrow: specificity of acetylcholinesterase staining
Authors: Lepore D. A. , Harris R. A. , Penington D. G.
Published: March 2008

Summary. Small cells staining diffusely for acetylcholinesterase, corresponding closely to the description of Jackson (19 73,19 74), have been quantitated in the bone marrow of mouse and rat. These cells fulfil the requirements of megakaryoblast precursors, showing elevation in numbers prior to megakaryocytes in response to thrombocytopenia, and carrying platelet‐specific antigens demonstrated by immunofluorescence. Some show a clearly defined nucleus whilst in others the nucleus is obscured; the femoral content of both types follows the same time course after acute thrombocytopenia and the latter can be distinguished from megakaryocyte cytoplasmic fragments.

Other small cells present in rodent bone marrow contain granular positivity after acetylcholinesterase staining but also granules staining with Luxol‐Fast‐Blue, suggesting that these cells are eosinophils. These cells are greatly increased in number with stimulation of eosinophilia by parasite infestation but show no response to acute thrombocytopenia. Peripheral blood eosinophils exhibit both acetylcholinesterase and Luxol‐Fast‐Blue staining.

A number of properties ascribed to granular‐staining small acetylcholinesterase‐positive cells previously presumed to be megakaryoblast precursors will need to be reassessed.
Myelokathexis: a rare form of chronic benign granulocytopenia
Authors: Bassan Renato , Viero Piera , Minetti Bruno , Comotti Benedetto , Barbui Tiziano
Published: March 2008

A case of childhood chronic neutropenia due to myelokathexis is reported. This extremely rare condition (the fourth case in the literature) has been diagnosed from the characteristic bone marrow abnormalities in the neutrophils.

Myelokathexis (kathexis‐retention) is a form of childhood granulocytopenia characterized by marked degenerative changes in mature neutrophils and considerable hyperplasia of the bone marrow myeloid cell series. The disease is rare and, since the first description by Zuelzer in 1964, only two additional cases have been reported (Krill et ah 1964; O'Regan et al 1977). We wish to describe the first case observed in Italy which is also the first reported outside U.S.A.
Preferential phagocytosis of in vivo aged murine red blood cells by a macrophage‐like cell line
Authors: Walker William S. , Singer Judith A. , Morrison Martin , Jackson Carl W.
Published: March 2008

The ability of an established line of mouse macrophages (IC‐21) to ingest red blood cells (RBC) aged in vivo was assessed. RBC populations of increasing age were prepared in mice by serial hypertransfusion, a procedure that inhibits erythropoiesis. Mouse RBC with a mean age of about 58 d (normal RBC life span, 60 d) had a circulating half‐life of less than 1 d when transfused into normal mice. IC‐21 macrophages ingested the in vivo aged RBC in preference to RBC from normal mice (mean RBC age, 30 d). RBC isolated from mice 10 d after being released from one red blood cell lifespan (60 d) of inhibited erythropoiesis (mean RBC age 5 d) were ingested significantly less than RBC from normal mice. The IC‐21 macrophage line used with in vivo aged RBC affords a highly defined model system for identifying the mechanism(s) of macrophage‐mediated homeostasis.
Plasmodium falciparum takes up iron from transferrin
Authors: Pollack Simeon , Fleming John
Published: March 2008

Plasmodium falciparum, cultured in vitro, is inhibited by desferrioxamine, a specific iron chelating agent, despite the abundant haem iron which surrounds the parasite. In this report we show that the intraerythrocytic parasite obtains iron from transferrin. The parasite may therefore be freed of a requirement to catabolize haem to obtain iron thus minimizing haem toxicity.
The effects of therapeutic dialysis and renal transplantation on uraemic serum inhibitors of erythropoiesis in vitro
Authors: Summerfield Geoffrey P. , Bellingham Alastair J.
Published: March 2008

The methyl cellulose culture system using human bone marrow and erythropoietin (Ep) at different concentrations was used to examine the effects of uraemic sera on erythropoiesis in vitro.

Sera from undialysed patients with advanced uraemia (plasma creatinine ± 900 μmol/l) when added to cultures at 10% with Ep at 2‐0 u/ml were consistently inhibitory to the growth of erythroid burst‐forming units (BFU‐E). No inhibition of erythroid colony‐forming units (CFU‐E) was observed at this Ep concentration but inhibition was consistently demonstrated with Ep at 0‐2 u/ml. Sera from undialysed patients with less severe uraemia (plasma creatinine < 900 μmiol/1) were not inhibitory to BFU‐E or CFU‐E at Ep 2‐0 u/ml.

Sera from patients with stable, functioning renal transplants were stimulatory to erythropoiesis in vitro with Ep at 2 ‐0 u/ml. This finding is consistent with the normal or increased haematocrits often found following renal transplantation. Sera from patients on maintenance haemodialysis and continuous ambulatory peritoneal dialysis (CAPD) were not significantly different from normal in their effect on BFU‐E growth in vitro and were slightly but significantly stimulatory to the growth of CFU‐E. This suggests that these two forms of dialysis are equally effective in reducing the activity of uraemic inhibitors in serum and that inhibition of the marrow response to Ep and/or burst‐promoting activity (BPA) is unlikely to be a major factor in the continued anaemia of dialysis patients.
Characterization of an Indian (δβ)° thalassaemia
Authors: Wainscoat J. S. , Old J. M. , Wood W. G. , Trent R. J. , Weatherall D. J.
Published: March 2008

The molecular basis of δβ thalassaemia in an Indian family is shown here to be due to a previously undescribed deletion within the β globin gene complex. Starting 3 kilobases from the 3’ end of the Aγ gene, the deletion removes the δ and β globin genes and continues to an unknown extent in the 3’ direction. Heterozygotes for this deletion have about 25% Hb F with a Gγ: Aγ ratio of 70:30 while interaction with β+ thalassaemia results in the clinical picture of thalassaemia intermedia.
Prognostic significance of chromosome abnormalities in chronic lymphocytic leukaemia
Authors: Pittman Sally , Catovsky D.
Published: March 2008

Lymphocytes from 33 out of 63 patients with B‐cell chronic lymphocytic leukaemia (B‐CLL) were successfully stimulated for cytogenetic analysis by means of two B‐cell mitogens: pokeweed mitogen and lipopolysaccharide‐B, used after pretreatment of the cells with neuraminidase and galactose oxidase. All patients had abnormal clones in 30‐100% of the cells analysed. Chromosomes more frequently involved were Nos. 1,3, 6,11,12,13 and 14. The most common abnormality was a marker 14q+ (breakpoint 14q32) seen in 17 cases; trisomy 12 was observed in seven cases. A clinical scoring system was used to investigate the correlation of chromosome abnormalities with prognosis. The group with 14q+ was often associated with features of progressive disease, namely; prolymphocytoid or Richter transformation, refractoriness to therapy, high WBC and advanced staging. A significant difference in survival was observed between patients with 14q+ and the rest: median survival from diagnosis being 45 months and over 64 months, respectively (P<0‐05); when survival was calculated from the time of chromosome analysis the values were 8 months and more than 41 months, respectively (P < 0 01). It is suggested that 14q + is acquired during the evolution of CLL and that this development may be a key event in the clinical progression of B‐CLL. Other abnormalities, including trisomy 12, were not found to be associated with a worse prognosis.
Chemotherapy and autografting for chronic granulocytic leukaemia in transformation: probable prolongation of survival for some patients
Authors: Haines M. E. , Goldman J. M. , Worsley A. M. , McCarthy D. M. , Wyatt S. E. , Dowding C , Kearney L. , Th'ng K. H. , Wareham N. J. , Pollock A. , Galvin M. C. , Samson D. , Geary C G. , Catovsky D. , Galton D. A. G.
Published: March 2008

Between June 1977 and July 1983 51 patients with Ph1‐positive chronic granulocytic leukaemia (CGL) in transformation were treated either by chemotherapy or by chemoradiotherapy followed by autografting with haemopoietic stem cells collected from their peripheral blood at the time of diagnosis. Forty‐eight patients were restored to a second chronic phase. The median duration of survival after autografting was 26 weeks (range 2‐152 weeks). Twenty‐one patients with relatively long durations of second chronic phase were treated again by autografting as consolidation or when transformation recurred; this selected group of patients survived longer than the 30 patients treated by autografting only once (medians 52 v. 13 weeks respectively, P<0‐01). There was no significant influence of the patients’ age, splenectomy status, type of transformation, treatment pre‐autograft or number of nucleated cells autografted on the duration of survival. Three patients treated in myeloid blastic transformation were restored to partially Phnegative haemopoiesis. We conclude that this approach to the management of CGL in transformation can offer benefit for a minority of patients and that further chemotherapy and autografting for patients still in second chronic phase may be valuable.
Evaluation of serum deoxythymidine kinase as a marker in multiple myeloma
Authors: Simonsson B. , Källander C. F. R. , Brenning G. , Killander A. , Åhre A. , Gronowitz J. S.
Published: March 2008

Summary. A recently developed deoxythymidine kinase assay utilizing 125I‐iododeoxyuridine as substrate was used in an investigation of sera from 122 untreated patients with multiple myeloma. Most patients had slightly elevated or normal serum deoxythymidine kinase activity (S‐TK), although in some patients values increased by more than forty‐fold were found. S‐TK correlated with the haemoglobin level but did not correlate with sex, age, erythrocyte sedimentation rate, nor with the serum concentrations of creatinine, β2‐microglobulin, Ca or M‐component. The distribution of S‐TK values in IgG, IgA and pure Bence‐Jones myeloma did not differ significantly.

Patients with IgG and IgA myeloma excreting light‐chain immunoglobulin in the urine had significantly higher S‐TK than non‐excreters. There was a significant correlation between S‐TK values and tumour cell mass as determined by clinical staging. A high pretreatment S‐TK (> 5·1 units) also distinguished a group of patients with a significantly shorter survival time. Patients with no response to initial therapy had significantly higher S‐TK values than those who did respond. In longitudinal studies of 11 patients, S‐TK was found to increase when the disease became more aggressive.

The possibility of diagnosing disease progression at an early stage by an elevation of S‐TK is discussed.
Daunomycin administered by continuous intravenous infusion is effective in the treatment of acute nonlymphocytic leukaemia
Authors: Lewis Jerry P. , Meyers Frederick J. , Tanaka Leora
Published: March 2008

Summary. Anthracyclines given by continuous infusion, as opposed to bolus administration, are associated with a reduced incidence of cardiac and gastrointestinal toxicity. Our study was developed to test the antileukaemic effect of anthracyclines given as a continuous infusion. Nineteen sequential patients admitted for treatment of acute nonlymphocytic leukaemia (ANLL) were managed with a regimen utilizing a 3 d continuous intravenous infusion of daunomycin (DNM), and the complete response rate was similar to this institution's past experience with antileukaemic regimens employing 3 d bolus DNM. In our studies, infusion DNM was at least as myelotoxic and antileukaemic as bolus DNM; thus this method of administration should be explored further in remission‐induction regimens for ANLL.
Endogenous origin of microbiologically‐inactive cobalamins (cobalamin analogues) in the human fetus
Authors: Muir M. , Landon M.
Published: March 2008

Summary. The cobalamin content of placenta, maternal and fetal sera was assayed by methods which measure microbiologically‐active and microbiologically‐inactive cobalamins. Only microbiologically‐active cobalamins were present in the placenta but both types were present in both maternal and fetal circulations.

We conclude that only microbiologically‐active cobalamins cross the placenta and that the microbiologically inactive cobalamins in the fetus must arise by metabolism in situ.
Regeneration of phosphorylated metabolites in stored erythrocytes in an open perfusion system: studies using 31P NMR spectroscopy
Authors: Chapman B. E. , Kuchel P. W. , Lovric V. A. , Raftos J. E. , Stewart I. M.
Published: March 2008

Summary. Human erythrocytes were maintained at high haematocrit in a metabolically functional state for several hours in a thermodynamically open perfusion apparatus. The concentrations of ATP and 2,3‐bisphosphoglycerate (2,3‐DPG) and pH were continuously monitored before and after metabolic perturbations by using 31P NMR; the monitoring was achieved with a 31P flow‐through probe. Methylphosphonate was added to plasma perfusion medium as a phosphorus concentration standard and as a 31P NMR pH probe molecule. The rates of decline of ATP and 2,3‐DPG levels in fresh cells in a glucose‐free medium were measured as were the rates of reformation in response to a ‘rejuvenation’ medium. Also, rates of ATP and 2,3‐DPG synthesis during perfusion with Krebs bicarbonate–0·5 mmol/l glucose and perfusion with pooled plasma were measured in cells that had been previously stored at 4°C for 5 weeks.
Correction of anaemia following renal transplantation: serial changes in serum immunoreactive erythropoietin, absolute reticulocyte count and red‐cell creatine levels
Authors: Rejman A. S. M. , Grimes A. J. , Cotes P. Mary , Mansell M. A. , Joekes A. M.
Published: March 2008

Summary. Improvement of erythropoiesis following successful human renal transplantation in eight patients was monitored by sequential measurements of haemoglobin, red‐cell creatine, absolute reticulocyte count and estimates of serum immunoreactive erythropoietin (siEp). SiEp increased in five patients after transplant, in three cases almost immediately after a return to normal of plasma creatinine. The increase in siEp was followed by a rise in the absolute reticulocyte count and red‐cell creatine and finally by an increase in the haemoglobin level. As the haemoglobin approached normal levels a decline in the absolute reticulocyte count preceded a fall in siEp levels. Red‐cell creatine also fell, though more gradually than the reticulocyte count. Acute graft rejection (in two patients) was associated with a fall in siEp. Chronic rejection (in one patient) was associated with persistent increases in siEp, reticulocyte count and red‐cell creatine; this patient subsequently developed erythrocytosis.
Degradation of factor VIII coagulant antigen by proteolytic enzymes
Authors: Rick Margaret E. , Popovsky Mark A. , Krizek Dennis M.
Published: March 2008

Summary.The factors responsible for the lability of factor VIII coagulant activity (VIII:C) and factor VIII coagulant antigen (VIII:CAg) are poorly understood. In this study the VIII:C and VIII:CAg are studied after incubation with plasmin, trypsin or α‐chymotrypsin. Both isolated human VIII:CAg and VIII:CAg associated with factor VIII‐related antigen (VIII R:Ag) are evaluated. The antigenic sites of the VIII:CAg are somewhat more stable to the action of these enzymes than the functional activity, although both follow a generally parallel degradation. A biphasic decay curve is seen in the initial time points. No stabilization of the functional or antigenic reactivity is observed in the presence of the VIII R:Ag.

Lower concentrations of each enzyme cause an initial rise in the factor VIII:C in the presence of VIII R:Ag, but not in the isolated VIII:CAg. Higher concentrations of α‐chymotrypsin cause activation of VIII:C and a slight decrease in the VIII:CAg values in both preparations. These enzymes may play a modulating role in the coagulation cascade through the activation and degradation of VIII:C and VIII:CAg.
Inhibitory effect of PWM‐stimulated OKT4+ subsets on erythro‐, granulo‐ and megakaryocytopoiesis in vitro
Authors: Sakamaki Hisashi , Hamaguchi Hiroyuki , Furusawa Shimpei , Shishido Hideo
Published: March 2008

Summary. Normal human peripheral blood T cells and T‐cell subsets defined by monoclonal antibodies of the OKT series were pretreated with pokeweed mitogen (PWM). Their effects on the haematopoietic precursors, erythroid (BFU‐E, CFU‐E), granulocyte‐macrophage (CFU‐GM) and megakaryocyte (CFU‐M) colony forming cells were evaluated by coculture. While unstimulated T cells and T‐cell subsets enhanced growth of autologous blood BFU‐E, PWM‐stimulated T and OKT4+ cells suppressed it, also inhibiting proliferation of both autologous and allogeneic bone marrow BFU‐E, CFU‐E, CFU‐GM and CFU‐M. PWM‐stimulated OKT8+ cells had little effect on the growth of any of the precursors at the cell concentration at which growth was completely inhibited by PWM‐stimulated OKT4+ cells. Irradiation of T or OKT4+ cells with 3000 rad before PWM stimulation completely abrogated the inhibition. These observations might be related to the mechanism of pancytopenia in some cases of immune‐mediated aplastic anaemia.
Cephalosporin‐induced immune neutropenia
Authors: Murphy M. F. , Metcalfe P. , Grint P. C. A. , Green A. R. , Knowles S. , Amess J. A. L. , Waters A. H.
Published: March 2008

Summary. Neutropenia is an occasional complication of treatment with cephalosporin antibiotics. This report describes two patients who had neutropenia while receiving high doses of cephalosporins. The neutrophil counts returned to normal after stopping the drug, and cephalosporin‐dependent neutrophil antibodies were demonstrated in both cases, using the granulocyte immunofluorescence test. In one patient, the immune neutropenia appeared to be due to a drug adsorption mechanism similar to penicillin‐induced haemolytic anaemia, while an immune complex mechanism may have been involved in the second patient.
Plasma cell acid phosphatase score in multiple myeloma and related disorders
Authors: Hoffmann J. J. M. L. , Breed W. P. M. , Swaak‐Lammers N.
Published: March 2008

Summary. A detailed classification of plasma cells stained for acid phosphatase activity is introduced. With this method, patients with multiple myeloma, non‐myeloma gammopathies, reactive plasmacytosis and other diseases in which plasma cells are involved, were investigated. The results show that our method can discriminate between multiple myeloma and reactive plasmacytosis. The overlap between multiple myeloma and other monoclonal gammopathies is much smaller than observed in other studies. Surprisingly low levels of acid phosphatase activity were found in the cells from patients with lymphoplasmacytoid immunocytoma. It is concluded that the acid phosphatase score can be of value for studying disorders in which plasma cells are involved.
Placental transport of low molecular weight heparin in the pregnant sheep
Authors: Andrew M. , Boneu B. , Cade J. , Cerskus A. L. , Hirsh J. , Jefferies A. , Towell M. E. , Buchanan M. R.
Published: March 2008

Summary. Standard heparin, an effective treatment for antepartum thromboembolic disease, is thought to be safe for the fetus since it does not cross the placenta. Recently, a number of low molecular weight heparins have been prepared which have been shown to produce less bleeding than standard heparin for an equivalent antithrombotic effect in experimental animals. These observations suggest that the low molecular weight heparins may also provide superior antithrombotic therapy in antepartum thromboembolic disease. However, it is not known whether the low molecular weight heparins cross the placenta. To determine this, we examined the pharmacokinetics of 125I‐labelled standard heparin and a low molecular weight heparin, and their anticoagulant effects in mother and fetus, using a pregnant sheep model. Catheters were inserted into maternal and fetal femoral arteries at 108–119 d gestation (term: 147 d). 1–3 days later the mothers were given a bolus i.v. injection of 5000 anti‐Xa units of 125I‐labelled standard heparin or low molecular weight heparin, CY 222. Nine serial blood samples were collected over 4 h from both mother and fetus for measurements of radioactivity, anti‐Xa activity (chromogenic) and activated partial thromboplastin times. When therapeutic levels of standard and CY 222 heparins were achieved in the mother, there was no detectable radioactivity or anticoagulant effect in the fetus. We conclude that standard heparin and the low molecular weight CY 222 do not cross the placenta in the pregnant sheep.
Increased degranulation of human myeloperoxidase‐deficient polymorphonuclear leucocytes
Authors: Dri Pietro , Cramer Rita , Menegazzi Renzo , Patriarca Pierluigi
Published: March 2008

Summary. Myeloperoxidase (MPO)‐deficient neutrophils (PMN) released considerably more β‐glucuronidase, lysozyme and vitamin B12‐binding activities, when exposed to opsonized zymosan (STZ), than the normal counterpart. Release of the soluble enzyme lactate dehydrogenase was not appreciably changed over the incubation time with particles in either cell type. MPO‐deficient PMN and normal PMN ingested STZ particles at a similar rate at early times, but thereafter phagocytosis by MPO‐deficient PMN was significantly higher than that by normal PMN. The difference in degranulation between the two cell types greatly exceeded the difference in ingestion and was evident already at early phagocytosis times when no difference in phagocytosis was observed; this suggested that the higher degranulation in MPO‐deficient PMN was at least in part independent of the increased ingestion. This was confirmed by experiments with the soluble stimulant N‐formyl‐L‐norleucyl‐L‐leucyl‐phenylalanine (FNLLP). MPO‐deficient PMN and normal PMN exhibited a comparable respiratory burst when exposed to FNLLP plus cytochalasin B, but the defective cells released more azurophilic and specific granule markers than normal PMN. These results indicate that MPO‐deficient PMN degranulate more than normal PMN and suggest a role for MPO in the regulation of degranulation.
The effect of 1‐deamino 8 D‐arginine vasopressin (DDAVP) in a nonhaemophilic patient with an acquired type II factor VIII inhibitor
Authors: de la Fuente Beatriz , Panek Sophia , Hoyer Leon W.
Published: March 2008

Summary. DDAVP (1‐deamino 8 D‐arginine vasopressin) was administered on two occasions to a patient with an acquired factor VIII inhibitor. Because the patient's autoantibody was low titre (1.8 Bethesda units) and had type II kinetic properties, it was expected that endogenous factor VIII/VWF released by DDAVP would be inactivated at a slow enough rate to permit satisfactory haemostasis during dental procedures. In both instances there was a 7–9‐fold increase in factor VIII above the baseline level (13 units/dl), and factor VIII activity remained above 35 units/dl for 5 h after the infusion. The inhibitor could not be detected in his plasma during this period and the activated PTT was shortened into the normal range. Our experience with this patient suggests that DDAVP has a role in the management of some patients with low titre autoantibodies that inactivate factor VIII.
IgA inhibitor to factor VIII/von Willebrand factor
Authors: Gralnick Harvey R. , Flaum Morris A. , Kessler Craig M. , Zimbler Harvey , Coller Barry S.
Published: March 2008

Summary. A 60‐year‐old Black female presented with a haemorrhagic diathesis and an acquired factor VIII/von Willebrand factor (VIII/vWf) inhibitor. This inhibitor was classified as an IgA immunoglobulin and was active not only against factor VIII coagulant (VIII:C) activity but also against plasma von Willebrand factor (vWf). The purified IgA also interacted with normal platelets to inhibit ristocetin‐induced platelet aggregation (RIPA). In contrast, studies with haemophilia A plasma and platelets revealed that the inhibitor did not react significantly with these plasmas or platelets. The significant differences in the inhibition of vWf assay both of the plasma and the platelets of the haemophilia A patients suggests that part of the haemorrhagic diathesis may be related not only to the inhibition of VIII: C but also to interference with platelet function. In addition, these studies suggest that there may be significant differences in the factor VIII‐related antigen (VIII R: Ag) on platelets in haemophilia A patients compared to normal.
Distribution of glycoprotein IIb‐IIIa complexes in the surface membranes of human platelets and megakaryocytes
Authors: Hourdille Paquita , Benabdallah Schéhérazade , Belloc Francis , Nurden Alan T.
Published: March 2008

Summary. The distribution of glycoprotein (GP) IIb–IIIa complexes in the surface membranes of human platelets and megakaryocytes was investigated by transmission electron microscopy of cells that had been incubated with Fab fragments of a human alloantibody (IgG L) specific for the complex. Binding was visualized by a second antibody conjugated to peroxidase or adsorbed onto gold particles. Initial studies showed that the peroxidase reaction product and the gold particles were to be found at the outer surface of unactivated platelets. The occasional small cluster of particles was present. A positive reaction, more apparent with peroxidase labelling, was also seen in the channels of the platelet open canalicular system. Gold particles were abundant on the outer surface of mature megakaryocytes, and their distribution resembled that on unactivated platelets. As with platelets, peroxidaselabelled antibodies penetrated better, and revealed GP IIb–IIIa in the demarcation membrane system. A double immunofluorescence study, involving Fab fragments of IgG L and rhodamine‐conjugated antibodies to factor VIII R:Ag, demonstrated the presence of GP IIb–IIIa in megakaryocyte precursor cells. Our results show that the GPIIb–IIIa complex is present in megakaryocyte membranes and that it appears at the same time as the other platelet antigens.
Platelet associated immunoglobulins (PAIgG and PAIgM) in autoimmune thrombocytopenia
Authors: Hegde U. M. , Ball S. , Zuiable A. , Roter B. L. T.
Published: March 2008

Summary

An enzyme linked assay system was used to quantitate platelet associated IgM (PAIgM) in addition to platelet associated IgG (PAIgG) in normal subjects and in 145 patients with autoimmune thrombocytopenia (AITP). The mean PAIgM level in normals was 1.17 ng/106 platelets with a range of 0.01–2.45 ng (mean ± 2 SD). The corresponding PAIgG values as follows: mean 6.0 ng, range 2.0–10 ng/106 platelets. Elevated PAIgG was seen in 67.6% and abnormally raised PAIgM in 79.3% of patients. Both values were raised together in 57.2% and either elevated PAIgG or PAIgM in 89.7%. All patients with PAIgG values greater than 4 times upper limit of normality were found to have abnormal PAIgM. The relevance of elevated PAIgM, the possible interaction between PAIgG and PAIgM and the implication of our results in patients with autoimmune thrombocytopenia are discussed.
Assay of factor VII activity by two techniques: evidence for increased conversion of VII to αVIIain hyperlipidaemia, with possible implications for ischaemic heart disease
Authors: Miller G. J. , Walter S. J. , Stirling Y. , Thompson S. G. , Esnouf M. P. , Meade T. W.
Published: March 2008

Summary

Factor VII was assayed in healthy adults and pregnant women by a coagulation method (VIIc) and a procedure (VIIt) based upon activation of factor X. Although VIIcand VIItwere highly correlated (r0.8) they apparently measured different aspects of VII activity. This difference was related to plasma lipid concentrations. Plasma VIIcshowed independent positive associationsin vivowith VIIt, cholesterol and triglyceride concentrations, but was unaffected byin vitroadjustment of plasma lipoprotein concentrations. The difference between assays might be due to differing reactivities of VII. The VIIcassay measures VII in itsin vivoproportions as the single‐chain protein and fully active double‐chain form (αVIIa). In VIIt, all VII is converted to αVIIabefore measurement. Thus an increase in VIIcbut not VIItwith increasing lipid concentrations reflects an increased proportion of VII as αVIIa, possibly secondary to activation of the contact system. This effect may explain at least part of the increased VIIcand normal VIItin pregnancy, and the increased VIIcof hyperlipidaemias in general. The relative values of VIIcand VIItare proposed as a measure of flux within the coagulation system, and as a measure of coagulability in hyperlipidaemia and other states.
Five haplotypes in Black β‐thalassaemia heterozygotes: three are associated with high and two with low Gγ values in fetal haemoglobin
Authors: Harano T. , Reese A. L. , Ryan R. , Abraham B. L. , Huisman T. H. J.
Published: March 2008

Summary

Genotypes at seven different polymorphic restriction sites (5’to the ° gene, at the Gγ, at the Aγ, at the Ψβ, 3’to the Ψβ, at the β, and 3’to the β genes) were analysed by restriction endonuclease mapping of the DNA from 66 Black β‐thalassaemia heterozygotes from Georgia and several of their normal relatives. Five different haplotypes were observed. Three of these were associated with high Gγ values in the small amount of Hb F (0.8‐8.3%) present in the blood of these patients and two with low Gγ values. One haplotype [‐ + ‐ ++++] that occurred on two of every three β thalassaemia chromosomes was associated with high Gγ levels, and is the same as that found in some Black SS patients also having high Gγ values (Gilman & Huisman, 1984). Two others [‐ ++ ‐ + ‐ +] and [−+−−+++] were also associated with high Gγ, while two [−−−−+++] and [+−−−−++] were associated with low Gγ. Variation in haematological data, mainly MCV and MCH values, was found to be caused in part by the type of β‐thalassaemia (defined by its haplotype) and by the presence of an additional α‐thalassaemia‐2 heterozygosity or homozygosity.
Fluctuating deformability of oxygenated sickle erythrocytes in the asymptomatic state and in painful crisis
Authors: Lucas G. S. , Caldwell N. M. , Stuart J.
Published: March 2008

Summary

A new gravity filtration technique for the study of sickle cell deformability has shown a significant loss of filterability of oxygenated erythrocytes on day 2 of painful crisis in eight patients with homozygous sickle cell anaemia; there was no increase in irreversibly sickled cell (ISC) count. Serial study (mean of 5.2 occasions) in six out‐patients when asymptomatic also showed considerable fluctuation in sickle cell filterability with time; this was again independent of ISC count. Erythrocyte filtration methods demonstrate fluctuations in the rheology of sickle cells that are not detected by ISC counts and which are probably secondary to clinically occult, as well as overt, vaso‐occlusive events in the microcirculation.
Detection of metastatic tumour cells in routine bone marrow smears by immuno‐alkaline phosphatase labelling with monoclonal antibodies
Authors: Ghosh A. K. , Erber W. N. , Hatton C. S. R. , O'Connor N. T. J. , Falini B. , Osborn M. , Mason D. Y.
Published: March 2008

Summary. The present study describes 11 cases (10 carcinomas, one rhabdo‐myosarcoma) in which immuno‐alkaline phosphatase labelling with monoclonal antibodies was used to demonstrate metastatic cells in routine smears of aspirated bone marrow. Carcinoma cells were detected using antibodies against epithelial cytokeratins, milk fat globule membrane antigen and carcinoem‐bryonic antigen, and rhabdomyosarcoma cells with monoclonal anti‐desmin. In four of the carcinoma cases it had not been possible to identify malignant cells in routinely stained marrow smears, whilst the case of disseminated rhabdomyosarcoma had initially been diagnosed (and treated) as a case of acute lymphoblastic leukaemia. The anti‐cytokeratin antibody was found to be the most valuable of the anti‐epithelial reagents used, since it labelled malignant cells in all of the 10 cases of carcinoma and gave the strongest reactions. These results suggest that immunocytochemical labelling should be used in cases of suspected carcinoma whenever conventional examination of marrow smears yields negative results, and furthermore (as illustrated by the case of rhabdomyosarcoma) that the technique is of value for identifying the true nature of poorly differentiated neoplasms in bone marrow.
Age dependent behaviour of red cell glycolytic enzymes in haematological disorders
Authors: Jansen G. , Koenderman L. , Rijksen G. , Punt K. , Dekker A. W. , Staal G. E. J.
Published: March 2008

Summary. The age dependent behaviour of 11, mainly glycolytic, red blood cell enzymes in 26 patients with various haematological disorders has been investigated after separation of red blood cells by discontinuous density gradient centrifugation. The frequency of enzyme deficiencies in the old cells of these patients was significantly increased in comparison with the unseparated cells, 29 and 13 deficiencies, respectively. Particularly hexokinase activity, although normal or even increased in unseparated cells, was found deficient in old cells in seven cases. In addition, an increased number of phosphofructokinase deficiencies was observed in the patients’old cells (eight cases) as compared to the unseparated cells (three cases). However, the red blood cells of the majority of these patients were found to contain increased enzyme activities, irrespective of cell age. Enzyme activities in the youngest cell population did not correlate with the reticulocyte count. Cases of high pyruvate kinase and hexokinase activities were studied for kinetical, electrophoretical and immunological properties of the respective enzymes, but no abnormalities could be demonstrated, indicating an increased synthesis of these enzymes.
Sequential development of distinct clonal chromosome abnormalities in a patient with preleukaemia
Authors: Miller Barbara A. , Weinstein Howard J. , Nell Marilyn , Henkle Carol T. , Dillon Patricia L. , Tantravahi Ramana
Published: March 2008

Summary Preleukaemia has been identified as a clonal haemopathy in which progression to acute leukaemia involves conservation of the preleukaemic karyotype in the blast cells or the development of new abnormalities superimposed on the original clone. In this report, a case of childhood preleukaemia is presented in which two cytogenetically distinct clones developed over 2 years in a dysplastic marrow that was initially karyotypically normal. One clone with 47 chromosomes (47, XY, + 21), disappeared without therapy. Predominance of the cytogenetically abnormal clone, 45,XY,−12,−17,t(12;17)(p11;q11) was associated with the development of acute myelogenous leukaemia and myelofibrosis. The development of independent clonal abnormalities in the unstable preleukaemic marrow may occur more commonly than has been previously recognized. Implications of the progression of the karyotypic abnormalities are discussed.
Differential effect of isolated placental isoferritins on in vitro T‐lymphocyte function
Authors: Matzner Y. , Konijn A. M. , Shlomai Z. , Ben‐Bassat H.
Published: March 2008

Summary The effect of isoferritins isolated from human term placenta on certain T‐lymphocyte parameters was studied in vitro using normal human lymphocytes. These isoferritins differed in ion exchange affinity, isoelectric point, and subunit composition. Only the acidic isoferritins caused a marked suppression of phytohaem‐agglutinin (PHA) blastogenesis and the most acidic isoferritin (‘Acid I’) was suppressive at a concentration as low as 0.25 μg/ml. All four isoferritins suppressed concanavalin A (Con A) blastogenesis in a similar concentration dependent manner, with maximum effect at an isoferritin concentration of 1 μg/ml.

Both basic and acidic isoferritins reduced the Con‐A‐capping phenomenon in normal lymphocytes at concentrations higher than 0.5 μg/ml, but at 0.25 μg/ml only the acidic isoferritin was effective. The above findings support our previous report concerning the suppressive effect of splenic ferritin on T‐lymphocyte function in vitro and indicate that acidic isoferritins, which often predominate in malignancy, demonstrate a higher degree of immunosuppressive activity. Thus, acidic isoferritins may play a role in the development of abnormal lymphocyte function encountered in certain proliferative disorders.
The flow of blood cell suspensions through 3 μm and 5μm Nuclepore membranes: a comparison of kinetic analysis with scanning electron microscopic examinations
Authors: Jones J. G. , Holland B. M. , Humphrys J. , Wardrop C. A.
Published: March 2008

Summary This study was designed to investigate the effect of red and white cells on the flow of dilute suspensions of blood cells through 3 μm and 5 μm Nuclepore membrane filters.

The rate of flow of blood cell suspensions through 3 μm or 5 μm membranes declines continually due to occlusion of pores by slow cells. The cells which occlude 3 μm pores exceed the number of white cells about seven‐fold. Electron microscopic examination of a used membrane confirms that these slow cells are not white cells but are red cells which are visibly damaged. With 5 μm membranes, the number of slow cells is entirely consistent with them being white cells. Again, electron microscopic examination confirms that 5 μm pores are occluded by white cells.

With both types of membrane, the same kinetic analysis is valid and yields information about the behaviour of red cells during the filtration procedure.
Life expectancy of Swedish haemophiliacs, 1831–1980
Authors: Larsson S. Anders
Published: March 2008

Summary Life expectancy of Swedish haemophiliacs has been estimated for the period 1831–1980. The data were derived from 948 haemophiliacs of whom 580 survived throughout 1980. Applying standard demographic techniques, median life expectancy of patients with severe haemophilia was found to have increased fivefold, from a mere 11 years during the period 1831–1920 to 56.8 years during 1961–80. The corresponding estimates for patients with moderate haemophilia were 27.5 and 71.5 years, respectively. The limited data on patients with mild haemophilia did not indicate any significant improvements in mortality. From the beginning of this century to 1980 median life expectancy for Swedish males increased from 61.7 years to 75.6 years, an increase of 23%. Analysing the last 12 years of the study (1969–80), death rates for patients with severe haemophilia below the age of 45 were not much different from those of Swedish males in the population as a whole. The investigation implies that the mortality of haemophiliacs in Sweden is approaching that of the total male population.
Studies of platelets from patients with the grey platelet syndrome
Authors: Greenberg‐Sepersky Sheryl M. , Simons Elizabeth R. , White James G.
Published: March 2008

Summary The grey platelet syndrome is a rare inherited disorder characterized by a marked decrease or absence of α‐granules and of platelet‐specific α‐granule proteins. By utilizing platelets from two patients with this syndrome, we here demonstrate that the initial response of human platelets to α‐thrombin does not require the presence of α‐granules nor the effective release of their constituents. Furthermore, these platelets respond to thrombin with a normal, dose‐dependent membrane potential change, and a normal secondary release of diS‐C3‐(5) thought to be released in parallel with β‐glucuronidase from the lysosomal granules. These results give new insight into the initial steps in the thrombin response of normal platelets.
Differences of immunoglobulin secreting cells in bone marrow from those in circulation: relevance to their development
Authors: Gyotoku Yuichi , Mori Mayumi , Nonaka Yasunobu , Nagata Yumiko , Saito Yoshimi
Published: March 2008

Summary The surface markers of immunoglobulin secreting cells (ISC) in bone marrow and peripheral blood were analysed. Circulating ISC bear surface Ig and Ia‐like antigens. However, these markers were not detectable on ISC in bone marrow. Fc and complement receptors were not present on circulating ISC. The areas of plaques corresponding to Ig secretion by bone marrow cells were always larger than those of peripheral blood cells. Although the majority of ISC were typical plasma cells, plasmacytoid lymphocytes were observed in peripheral blood. These findings seem to indicate that ISC in the peripheral blood are less advanced in their differentiation and maturation pathway from B lymphocytes to plasma cells than those in bone marrow.

ISC in mesenteric lymph nodes exhibited nearly the same phenotype as peripheral cells.
Repopulating potential of canine bone marrow cells: differences between large and small cells separated by velocity sedimentation
Authors: Raghavachar A. , Prümmer O. , Calvo W. , Nothdurft W. , Steinbach K. H. , Fliedner T. M.
Published: March 2008

Summary

This study compares the pattern of haemopoietic recovery in dogs after total‐body irradiation and transfusion of different populations of cryopreserved autologous bone marrow cells. Dogs in group 1 received unseparated marrow cells. Group‐II dogs were transfused with small (<5.1 mmm/h) and group‐III dogs with large (>7.1 mm/h) bone marrow cells, separated by velocity sedimentation. Myeloid progenitor cells (CFU‐GM) present in slowly sedimenting cell fractions were characterized by a higher radiosensitivity in vitro and a lower proportion of cells in S‐phase as compared to rapidly sedimenting CFU‐GM. Although autografts in all groups contained comparable numbers of CFU‐GM, transfusions resulted in different patterns of recovery. Fractions of small bone marrow cells contained most of the pluripotent stem cells, leading to speedy haematological reconstitution and long‐term survival. The pattern of early recovery was similar in dogs of group I and of group II. In group III, the recovery in all cell lineages was delayed, going along with marked extramedullary haemopoiesis. The data may indicate the limits of committed progenitors in reflecting pluripotential stem cells, in particular, if bone marrow grafts have been modified in vitro. Differences in the repopulating potential of the graft might be reflected by distinct physical and biological properties of the CFU‐GM.
A prospective trial of young red cells in 48 patients with transfusion‐dependent thalassaemia
Authors: Marcus R. E. , Wonke B. , Bantock H. M. , Thomas M. J. G. , Parry E. S. , Taite H. , Huehns E. R.
Published: March 2008

Summary

Young red cells (YRBC) prepared on a cell washer were compared to whole blood in a randomized prospective trial in 48 transfusion dependent patients with thalassaemia major. A minor but statistically significant decrease in blood consumption was observed in the group receiving YRBC. However, no difference was seen in transfusion interval, mean haemoglobin and rate of haemoglobin fall between the two groups. Filtered blood was shown to be as effective as frozen blood in eliminating non‐haemolytic febrile transfusion reactions in all the trial patients. We conclude that any reduction in the rate of iron loading brought about by the use of YRBC is clinically insignificant and does not justify the expense, time and work required to produce young red cells for use in a large transfusion dependent thalassaemic population.