bluuebunny/crossref_metadata_2025 huggingface dataset:
Regulation of Human Bone Marrow Leucopoiesis
Authors: Golde D. W. , Cline M. J.
Published: March 2008

Summary. The clonal growth of bone marrow in soft‐gel culture is largely dependent on the presence of colony‐stimulating factors (CSF). However, when human marrow is cultured in liquid medium with the in vitro diffusion chamber technique, proliferation and differentiation of granulocytes and mononuclear cells proceeds without a requirement for exogenously supplied CSF. The addition of highly active monocyte‐conditioned medium to the liquid marrow cultures did not result in a measurable change in viable or differential cell counts. Similarly, when mono‐cytes were co‐cultivated with bone marrow in a double‐chamber apparatus, no stimulation of marrow cell growth was observed. Colony‐stimulating activity was identified in conditioned medium derived from the liquid marrow cultures, and the cultured marrow cells themselves stimulated colony formation when incorporated as feeder layers. Retention of marrow cell responsiveness to CSF in liquid culture was demonstrated by cloning the cultured marrow cells in agar at intervals up to 2 weeks. Evidence was obtained that bone marrow cells of the monocyte‐macrophage line were primarily responsible for the elaboration of CSF in liquid culture. These data suggest a role for humoral interactions between haematopoietic cells within the marrow cavity.
Liver Iron: Changes Induced by Cooking and Acid‐Peptic Digestion
Authors: Naish R. , Kimber C. L. , Deller D. J.
Published: March 2008

Summary. Guinea‐pig liver was used as a model for studies on the handling of dietary liver iron. Cooking and acid‐peptic digestion of the liver produced an increase in the low molecular weight liver iron complexes as demonstrated by dialysis and Sephadex gel filtration. Absorption studies in rats suggested that the reduction in the size of the iron complexes, following cooking and enzymatic digestion, increased the absorbability of the liver iron.
Haematopoiesis Measured by Spleen Colony and Diffusion Chamber Techniques in Mice Treated with One or Two Injections of Cyclophosphamide
Authors: Bøyum A. , Carsten A. L. , Laerum O. D.
Published: March 2008

Summary. The effect of cyclophosphamide (CY) on haematopoiesis in mice was observed by the techniques of the spleen colony and the diffusion chamber. Two hours after injection of CY, the number of spleen colony‐forming units (CFU) was reduced to 11% of control, whereas diffusion‐chamber progenitor cells (DCPC) were less affected and 30% survived initially. Histological examination showed that erythroid CFU were significantly more reduced than granuloid CFU, most likely because CY selectively impairs the ability of surviving multipotent stem cells to erythroid differentiation. This defect was repaired within a few days.

A second injection of CY 4 days after the first had similar effects, indicating that the selective effect on erythroid CFU was independent of proliferative rate. DCPC were less affected by the second CY injection than by the first, possibly because a higher proliferative rate increased their resistance towards CY.

The pattern of depletion and regeneration after one or two injections of CY indicates that the two techniques do not assay identical cell populations. Whereas the spleen colony technique predominantly measures multipotent stem cells, the diffusion chamber technique most probably to a larger extent measures unipotent (committed) stem cells.
Effect of Heterologous Anti‐lymphocyte and Anti‐granulocyte Sera on Haematopoietic Progenitor Cells in the Rat
Authors: Benestad H. B. , Iversen J.‐G. , Warhuus K.
Published: March 2008

Summary.

The effect on haematopoietic progenitor cells of specific antisera against mature haemic cells has been investigated.

Heterologous anti‐lymphocyte and anti‐granulocyte sera were prepared by injecting suspensions of rat target cells into rabbits. Recirculating lymphocytes were obtained from thoracic duct lymph, non‐recirculating ones from blood of rats in which thoracic duct lymph had been drained away for 3 days, and mature granulocytes were isolated from blood of normal rats. Preliminary assaying of the cytotoxic activity of the sera was done by measuring isotope release from target cells proper, labelled with 51 Cr. When absorption procedures had been carried out the antisera were apparently specific for recirculating lymphocytes, nonrecirculating lymphocytes, or granulocytes.

The effect of these antisera on progenitor cells for granulocytes and on mature cell types were then evaluated with the diffusion chamber culture technique. Mononuclear cells from rat blood, bone marrow cells, or a mixture of both were incubated with different dilutions of antisera before culturing for I or 4 days. The yield of the cell types used for immunization was reduced with all employed dilutions of antisera. The number of proliferative granulocytes harvested from 4‐day cultures was taken to reflect the number of progenitors inoculated. Anti‐recirculating lymphocyte sera never affected progenitor cells, and they could be spared with some dilutions of antisera against nonrecirculating lymphocytes and granulocytes.

It is thus possible to prepare specific antisera against lymphocytes with no harmful effect on progenitor cells.
Renal Fibrin Deposition Associated with Isoimmune Haemolysis in Monkeys: Observation by Light, Electron and Immunofluorescence Microscropy
Authors: Roth G. J. , Poskitt T. R. , Bobrow J. C. , Fortwengler H. P.
Published: March 2008

Summary.

Following control renal biopsies, five cynomolgus monkeys received an infusion of incompatible isoimmune plasma. Renal biopsies were obtained at 2 or 4 hr, 24 hr, and 7 days after infusion and were examined by light and electron microscopy, as well as immunofluorescence microscopy for platelet antigens and fibrinogen related antigen(s) (FRA). Test animals receiving incompatible plasma showed evidence of haemolysis and disseminated intravascular coagulation. Renal tissue changes varied according to the severity of the transfusion reaction and the time of biopsy. Light microscopy revealed mild glomerular and tubular changes. In early biopsies, immunofluorescence studies showed deposition of FRA in glomerular capillaries and tubule cells. By day 7, glomerular FRA was no longer present, and tubule cell FRA had been displaced into the renal interstitium. Platelet antigen was not seen in any biopsy. Electron microscopy demonstrated proximal tubule cell damage with phosphotungstic acid positive staining granules, intravascular clot formation, and foot process fusion. Two control monkeys received compatible plasma and showed no evidence of renal injury. It appears that renal fibrin deposition and tubule cell damage occur even during mild haemolytic transfusion reactions. This study provides evidence for local fibrinolysis of glomerular fibrin and the transfer of FRA from tubular lumens to the interstitium. In addition, tubule cell FRA appears to be associated with structural damage.
Studies in Lipogeiiesis by Red Cells of Mice
Authors: Pushpendran C. K. , Eapen Jacob
Published: March 2008

Summary. The incorporation of[1‐14C]acetate,[U‐14C]glucose and [1‐14C]palmitate into lipids of red cells from 5‐, 30‐ and 180‐day‐old mice has been studied. The results show that the cells from 5‐ and 180‐day‐old mice have greater capacity for lipid synthesis than those from 30‐day‐old animals. Ghost preparations exhibit a similar pattern of incorporation but the levels of incorporation are higher than those of intact cells. There arc age‐dependent differences in the incorporation of the precursors into various types of lipids. The results show that there are changes in the levels of free fatty acids and triglycerides of red cells as a function of animal age. No quantitative changes were observed in phospholipids and cholesterol.
Procarbazine Induced Oxidative Haemolysis: Relationship to in Vivo Red Cell Survival
Authors: Sponzo R. W. , Arseneau J. C. , Canellos G. P.
Published: March 2008

Summary. A new hydrazinc derivative, N‐isopropyl‐α‐(2‐methyl‐hydrazino)‐p‐toluarnide hydrochloride (MIH, Procarbazine, Natulan), has been used widely in the treatment of neoplastic disease. When in vitro incubations of normal and C6PD‐deficient red cells with procarbazine were performed, oxidative changes in red cells were detected similar to those found with other hydrazine derivatives. These changes consisted of an increased osmotic fragility, decreased levels of reduced glutathione (GSH), increased percentages of methaemoglobin, and the generation of intracellular hydrogen peroxide. G6PD‐deficient red cells appeared more susceptible to these effects. When the known degradation products of procarbazinc metabolism were studied, the hydrazone was found to be the most potent oxidant while the other metabolites exhibited only minimal activity. Since the potential for in vivo haemolysis in patients receiving large doses of parenteral procarbazinc was suggested, the 51Cr survival of G6PD deficient red cells was performed in two patients receiving procarbazinc (1.5 g/m2, intravenously). No decrease in the red‐cell survival was detected.
Experimental Hepatic Porphyria Induced by Hexachlorobenzene as a Model for Human Symptomatic Porphyria
Authors: Stonard M. D.
Published: March 2008

Summary. Chronic feeding of hexachlorobenzene to rats increases the levels of certain hepatic haemoproteins prior to the onset of hepatic porphyria. The onset of hepatic porphyria is slow and only water soluble porphyrins accumulate in the livers. The accumulation of hexachlorobenzene by the liver is progressive but no direct correlation exists between the amount of HCB in the liver, the extent of hepatic porphyria and the changes in haemoprotein concentration. Similarities between experimental hepatic porphyria induced by hexachlorobenzene and those seen in human symptomatic porphyria are presented and emphasize the usefulness of the hexachlorobenzene model with which to study the human disease.
Ferritin in Blood Cells from Normal Subjects and Patients with Leukaemia
Authors: Worwood M. , Summers M. , Miller F. , Jacobs A. , Whittaker J. A.
Published: March 2008

Summary. Normal human leucocytes contain about 7 fg ferritin protein per cell. In 16 patients with acute myeloblastic leukaemia the mean concentration was 46 fg per cell, in acute myelomonocytic leukaemia 175 fg per cell, and in chronic granulocytic leukaemia 16 fg per cell.

It is suggested that the high serum ferritin concentration in leukaemic patients is derived from the leukaemic cells themselves and that it may be due to increased synthesis.
The Effects of Inhibitors of Microtubule and Microfilament Function on Transferrin and Iron Uptake by Rabbit Reticulocytes and Bone Marrow
Authors: Hemmaplardh Deeporn , Kailis S. G. , Morgan E. H.
Published: March 2008

Summary. The possible involvement of microtubules and microfilaments in the uptake of transferrin and iron by rabbit reticulocytes and bone marrow cells was investigated by using agents which interfere with the functions of these cellular components. Colchicine, vinblastine, vincristine, strychnine, and heavy water, which inhibit microtubule function, all diminished transferrin and iron uptake by the cells. Vinblastine was also shown to inhibit transferrin release fiom reticulocytes and the uptake of both citrate‐bound iron and leucine. Cellular ATP levels were not affected. Cytochalasin B which is believed to disrupt microfilaments did not affect transferrin or iron uptake by the cells. It is concluded that the inhibitory effects observed were due to alteration of structural components of the cell membranes, probably in the microtubules.
Storage Iron Kinetics: VI. THE EFFECT OF INFLAMMATION ON IRON EXCHANGE IN THE RAT
Authors: Hershko Chaim , Cook James D. , Finch Clement A.
Published: March 2008

Summary. The effect of turpentine‐induced inflammation on iron exchange in the rat has been studied. Within a few hours the basal level of plasma iron turnover fell from 1539 to 430 μg/kg/day, and iron absorption from the gut was reduced from 910 to 235 μg/kg/day. Early release of iron from nonviable red cells by reticulo‐endothelial cells was reduced from 72% to 38%, whereas early release of haemoglobin iron by hepatocytes was reduced from 29% to 17% with inflammation. These observations provide quantitative definition of the effects of inflammation on iron nietabolism and further demonstrate its general effect in impairing iron exchange by a number of cellular transport systems.
Hepatocellular Uptake of Ferritin in the Rat
Authors: Unger Arie , Hershko Chaim
Published: March 2008

Summary. Over 85% of ferritin 59Fe is recovered in the liver of rats within 24 hr of its intravenous injection. Autoradiographic studies have shown that this hepatic uptake is restricted entirely to the parenchymal cells. Following its hepatocellular uptake ferritin is catabolized and its iron moiety is recombined with locally produced apoferritin. The rate of hepatic uptake is reduced by bleeding and enhanced by inflammation. The electrophoretic properties and hepatic handling of ferritin preparations derived from the liver (parenchymal) or spleen (reticuloendothelial) are identical. These findings suggest that the increased amounts of circulating ferritin found in pathologic conditions such as transfusion haeniosiderosis and ineffective erythropoiesis, might play a significant role in the production of parenchymal iroll overload.
Granulocyte Colony Stimulating Activity and Vitamin B12 Binding Proteins in Human Urine
Authors: Gibson Emma L. , Herbert Victor , Robinson William A.
Published: March 2008

Summary. The levels of granulocyte colony stimulating factor (CSF) in the urine of normal humans and patients with various diseases have been compared with levels of urinary B12 binding proteins (BBP). A correlation has been shown between the levels of these parameters in some urines collected at random from patients with a wide variety of diseases. Serial urine samples from two patients who received injections of vitamin B12 also showed a correlation between rises and falls of CSF and BBP over a period of several days. Biochemical similarities exist between these factors and both appear to be, at least in part, secreted by mature white blood cells as the result of an active metabolic process. These findings suggest that CSF and BBP may be closely related and perhaps the same.
Neutrophil Function in Chronic Granulocytic Leukaemia before and after Busulphan Treatment
Authors: Whittaker J. A. , Khurshid M. , Hughes H. R.
Published: March 2008

Summary. Neutrophil mobilization, phagocytic index (PI) and nitroblue tetrazolium (NBT) dye reduction were reduced in 12 untreated patients with chronic granulocytic leukaemia whilst the serum lysozyme (SL) was increased. After nine patients had been treated with a standard dose of busulphan neutrophil mobilization was normal in six patients and NBT dye reduction normal in all nine patients but although the PI rose towards normal a slight defect which was possibly due to Busulphan therapy remained. The SL fell towards normal in nine patients after treatment but remained significantly different from a control group.
The Effect of Platelet Membrane Antibodies on Aggregation and Release
Authors: Kaplan K. L. , Nachman R. L.
Published: March 2008

Summary. Human platelets were preincubated with Fab fiagments derived from two types of platelet antibodies: (1) antibody to whole platelet membranes, and (2) antibody to the major platelet membrane glycoprotein. Subsequently platelet aggregation and release of [14C]serotonin in response to adenosine diphosphate (ADP), adrenaline, thrombin, and collagen were evaluated. Following incubation with the anti‐membrane antibody, significant inhibition of both aggregation and serotonin release was observed. The pattern of inhibition of ADP‐ and thrombin‐induced aggregation and release differed from the inhibition noted after the addition of adrenaline or collagen. Treatment of platelets with the anti‐membrane glycoprotein antibody had no effect on subsequent aggregation induced by ADP, adrenaline, thrombin or collagen.
Factor VIII of Small Molecular Weight and its Aggregation
Authors: Austen D. E. G.
Published: March 2008

Summary. Work on the disaggregation of factor VIII into sub‐units has been extended and it is shown that factor VIII of small molecular weight may be aggregated to a high molecular weight species probably identical to the nod unreacted factor. In addition some practical modifications were introduced to improve yield and reproducibility. The reaction of dithiothreitol and high concentrations of saline were compared using both human and bovine factor VIII. Small factor VIII which was produced had a molecular weight of 230 000 and the act of disaggregation illustrated a clear difference in behaviour between active factor VIII and factor VIII related antigen. These results may be considered to oppose the hypothesis that factor VIII related antigen is the protein carrier of factor VIII.
The Anti‐Rho(D) Responses of Immunized Volunteers following Spaced Antigenic Stimuli
Authors: Gunson H. H. , Stratton F. , Phillips P. K.
Published: March 2008

Summary. Twenty‐six Rho(D) negative male volunteers previously immunized with Rho(D) positive red cells have received spaced antigenic stimuli in an attempt to produce high serum anti‐D activity during the secondary responses.

Although there is a considerable individual variation in magnitude, there is, in general, either a rapid or slow fall from the maximum anti‐D concentration achieved to some steady state of anti‐D concentration which persists with little change for several months.

A case is made for the acceptance of an anti‐D concentration of 20 International Units (i.u.) per cm3 (about 7μ g/cm3) as the minimum requirement for accepting plasma donations for immunoglobulin preparations. On this basis 19 of the 26 volunteers could contribute regularly, although in only 10 volunteers was a concentration of 100 i.u./cm3 or more maintained for periods of several months following an antigenic stimulus.

No correlation was observed between the maximum concentration achieved in the primary response and that at the steady state following further antigenic stimulation. The potential of a volunteer cannot, therefore, be predicted from the early immune response. Indeed with certain volunteers after regular spaced antigenic stimuli, markedly reduced antibody responses have been observed.

The implications of these findings are discussed with respect to the collection of suitable donations of anti‐D plasma.
The Effect of Splencetomy on the Haematological Response to Radiotherapy in Hodgkin's Disease
Authors: Begent R. H. J. , Wiltshaw E.
Published: March 2008

Summary. Splenectomy performed as part of the initial treatment of Hodgkin's disease resulted in less depression of platelet count during radiotherapy than that seen in patients who had radiotherapy followed by splenectomy. The difference was Significant whether the removed spleen was grossly enlarged or not. There was no significant difference in white cell count in the two groups, nor was the duration of radiotherapy longer in the patients who had splenectomy after radiotherapy. We concluded that splencetomy as part of the initial treatment does not confer any worthwhile protection to the bone marrow during mantle radiotherapy.
Oxygen Binding to Haemoglobin in Subjects with Hypoproliferative Anaemia, with and without Chronic Renal Disease: Role of pH
Authors: Lichtman M. A. , Murphy M. S. , Whitbeck A. A. , Kearney E. A.
Published: March 2008

Summary. The relationships of haenioglobin concentration and blood pH to red cell 2,3‐dipliosphoglycerate and oxygen binding by haenioglobin have been studicd in healthy subjects and subjects with hypoproliferative anaemia with or without severe chronic renal disease. Red cell 2,3‐DPG was inversely correlated with haemoglobin deficit and directly and equally strongly associated with blood pH in anaemic subjects without chronic renal disease. In subjects with chronic renal disease receiving regular haemodialysis, predialysis pH was not increased despite severe anaemia, and red cell 2,3‐DPG was not significantly elevated, except in subjects who had a sustained alkalosis due to the use of sodium bicarbonate.

In hypoproliferative anaemia, the incrcmcnt in pH was associated with the decrease in haenioglobin concentration such that 80% of the increase in p50 measured at standard conditions which occurred with anaemia was explicable by the relationship of (a) pH with haemoglobin concentration, (b) red cell 2,3‐DPG with pH, and (c) pso std with red cell 2,3‐DPG. However, p50 at the pH and base excess present in vivo was similar in all anaemic subjects whether an increase in red cell 2,3‐DPG occurred or not. Blood alkalosis and the accumulation of 2,3‐DPG cancelled each other's effect on oxygen binding by haemoglobin. Hence, increased red cell 2,3‐DPG and p50 compensated for the alkalosis of hypoproliferative anaemia, not for the deficit in haemoglobin concentration.
Human Isoferritins: Organ Specific Iron and Apoferritin Distribution
Authors: Powell L. W. , Alpert E. , Isselbacher K. J. , Drysdale J. W.
Published: March 2008

Summary. Ferritins from human liver, spleen, heart, pancreas and kidney were compared by electrophoresis and isoelectric focusing in polyacrylamide gels, by immunodiffusion against antisera to homologous and heterologous ferritin, and in some cases by their cyanogen bromide peptides. All ferritins appeared to consist of a single species on gel electrophoresis with the exception of heart ferritin which separated into two major components. Small differences in electrophoretic mobility were found in all tissue ferritins. By contrast, all tissue ferritins were found to consist of multiple forms when analysed by gel electrofocusing. At least five isoferritins were found in most tissues, several of which were common to most tissues. At least two were common to all tissues. Those ferritins which were most easily distinguishable electrophoretically, e.g. spleen and heart ferritin, showed the greatest differences on gel electrofocusing. The ferritin profile was characteristic of each organ and was reproducible both within individuals and between individual tissues. There were striking differences in the iron content of the various isoferritins within a tissue. Further, the iron content of isoferritins common to more than one tissue varied with the tissue of origin. Some isoferritins in several organs and all of the isoferritins in pancreas appeared to contain little, if any, iron. All five tissue ferritins contained antigenic determinants in common with liver ferritin. However, an additional antigenic determinant was found in liver ferritin which was not detectable in the ferritins from the other organs.
Ultrastructure of Lymphocyte‐Mediated Cytotoxicity
Authors: Tonietti G. , Pecci G. , Liberatore C. , Salsano F. , Fontana L.
Published: March 2008

Summary. The ultrastructural appearances of the interaction between normal phytohaemagglutinin (PHA)‐stimulated human lymphocytes and chicken red cells (ChRBC) were studied. Close contact between ChRBC and control non‐stimulated lymphocytes was often observed indicating that this is a non‐specific phenomenon. In mixtures of PHA‐stimulated lymphocytes and ChRBC, time‐dependent development of interdigitating pseudopods between the two cell types was observed which was not seen in controls. Their development followed the same time‐course as release of 51Cr when the ChRBC were pre‐labelled with this marker. The findings suggest a relationship between the morphological changes observed and the destruction of the target cells (ChRBC).
A New Assay for the Measurement of Total Progressive Antithrombin
Authors: Lane Janet L. , Bird Prudence , Rizza C. R.
Published: March 2008

Summary. A new rapid method for assaying total antithrombin activity has been developed based on the inactivation of thrombin incorporated into an agarose gel, during the radial diffusion of plasma in the gel. The area of thrombin inactivation is subsequently observed by the coagulation of fibrinogen in a separate agarose gel layer poured over the thrombin gel. The method is described in detail and its accuracy assessed with respect to other antithrombin assays. Using specific antisera to α2‐globulin (antithrombin III), α2‐macroglobulin and α1‐antitrypsin, total antithrombin activity measured by this assay consisted of 47%α2‐globulin, 29%α2‐macroglobulin and 26%α1‐antitrypsin.
Surface Features of Human Eosinophils: a Scanning and Transmission Electron Microscopic Study of a Case of Eosinophilia
Authors: Polliack Aaron , Douglas Steven D.
Published: March 2008

Summary. Cells were obtained from the peripheral blood of a patient with marked non‐leukaemic eosinophilia. Transmission electron microscopy (TEM) showed typical mature eosinophils. The surface architecture of eosinophils by scanning electron microscopy (SEM) shows that most were spherical with varying numbers of microvilli; a smaller proportion of cells had ridge‐like profiles, small ruffles and occasionally blebs. The surface features of the eosinophil thus resemble most lymphocytes and this finding must be considered when leucocyte populations are examined by SEM.
The Endotoxin‐induced Coagulant Activity of Human Monocytes
Authors: Rivers R. P. A. , Hathaway W. E. , Weston W. L.
Published: March 2008

Summary. Leucocyte suspensions, exposed to endotoxin in vitro, develop coagulant activity which has been identified as tissue factor. Pure suspensions of polymorphonuclear (PMN) neutrophils, lymphocytes and monocytes were exposed to endotoxin and tested for tissue factor activity after 4 h incubation at 37°C. The results indicate that the monocyte is the cell primarily responsible for the endotoxininduced coagulant activity of mixed leucocyte suspensions, the small amount of activity demonstrated in PMN neutrophil and in lymphocyte suspensions at high cell concentrations being accounted for by monocyte contamination of less than 1%.
Colony Formation by Human Haemopoietic Precursor Cells Cultured in Semi‐solid Agar in Diffusion Chambers
Authors: Gordon M. Y. , Blackett N. M. , Douglas I. D. C.
Published: March 2008

Summary.

A technique which allows colony growth of haematologically normal human bone marrow cells is described. The cells are supported by semi‐solid agar‐medium inside modified Millipore diffusion chambers implanted in the peritoneal cavity of irradiated mice. After 9 days incubation colonies containing up to 1000 cells are found in these Agar Diffusion Chambers. All haematologically normal patients studied so far produced colonies, the majority with between 10 and 40 colonies per 2 × 105 bone marrow cells inoculated.

This culture system therefore provides a convenient and reliable clonal assay for human bone marrow cells which, in contrast to the agar colony assay in vitro, does not require a source of Colony Stimulating Factor (CSF).
Serum Inhibitors of Fibrinolysis
Authors: Gallimore M. J.
Published: March 2008

Summary.

Four protease inhibitors have been identified in human serum and methods for their isolation are described. After removal of the euglobulin fraction, serum was submitted to ion exchange chromatography and gel filtration, and fractions were tested for inhibition of the lysis of plasminogen‐deficient fibrin clots by plasmin, trypsin and elastase. In addition inhibitors of plasminogen activation were sought by studying the effects of separated fractions on the lysis of plasminogenrich fibrin clots by urokinase. Examination by immunophoresis showed that three of the separated inhibitors were α2‐macroglobulin, α1‐antitrypsin and inter‐α‐trypsin inhibitor. The fourth antiprotease was a powerful inhibitor of both urokinase‐induced and plasmin‐induced clot lysis, and was identified as an inter‐α‐globulin from its electrophoretic mobility in agarose gels.
Portal Hypertension in Chronic Leukaemia
Authors: Datta D. V. , Grover S. L. , Saini V. K. , Datta B. N. , Aikat B. K. , Chhuttani P. N.
Published: March 2008

Summary.

Portal haemodynamic studies were carried out in 11 subjects, seven with chronic myeloid leukaemia and four with chronic lymphatic leukaemia, and results compared with those obtained in five patients with ‘idiopathic’splenomegaly and with control subjects.

All 11 patients with chronic leukaemia had intrasplenic pressures above 11 mmHg and of these four had pressures above 20 mmHg. Portosystemic collaterals were seen on splenovenography in four of these patients. Hepatic vein wedge pressure was above 7 mmHg in nine patients and these high levels were a result of increased free hepatic vein pressure. The corrected sinusoidal pressure and post sinusoidal resistance were essentially normal in all patients. Evidence of increased pre‐sinusoidal resistance was seen in six patients. Estimated hepatic blood flow above 1500 ml/min was seen in six patients and all had evidence of histological portal or sinusoidal infiltration.

Patients with ‘idiopathic’splenomegaly regardless of increased liver blood flow did not have a significant increase in intrasplenic pressure and no alteration in other haemodynamic parameters. It appears that increased liver blood flow alone in a normal liver can only minimally elevate intrasplenic pressure but can play a significant part in the pathogenesis of portal hypertension when liver structure is altered.
Ultrastructural Studies of the Bone Marrow in Sickle Cell Anaemia
Authors: Grasso J. A. , Sullivan A. L. , Sullivan L. W.
Published: March 2008

Summary.

Electron microscopic studies of bone marrow aspirates obtained from patients with homozygous sickle cell anaemia (HbSS) were fixed immediately without attempts to deoxygenate the samples. Erythroblasts and normoblasts in these preparations were devoid of haemoglobin polymers or other indications of sickling. Furthermore, the nucleated erythroid cells from sickle‐cell patients presented an ultrastructural morphology indistinguishable from that of identically‐processed erythroid cells in marrow samples from normal human volunteers. This report presents a description of the ultrastructural features of pronormoblasts and normoblasts in normal and sickle‐cell marrows and stresses the essentially normal appearance of nucleated erythroid elements in sickle cell anaemia. Exposure of sickle‐cell marrow aspirates to nitrogen at 37°C for 30 min resulted in haemoglobin polymerization in most erythrocytes and reticulocytes but only in 10–20% of the nucleated erythroid cells. Haemoglobin polymers in the form of intertwining tibre mesh works were observed in reticulocytes, orthochromatic and polychromatophilic normoblasts, but were absent in basophilic normoblasts and pronormoblasts. The results suggest that the concentration of haemoglobin in intramedullary normoblasts may be the limiting factor determining the predisposition of these cells to undergo sickling as well as the pattern of haemoglobin aggregation. Under the physiological conditions prevailing in the marrow, haemoglobin concentration in normoblasts may be insufficient to result in aggregation and polymerization.
Splenectomy in Homozygous Beta Thalassaemia: A Retrospective Study of 30 Patients
Authors: Engelhard Dan , Cividalli Gabriel , Rachmilewitz Eliezer A.
Published: March 2008

Summary.

In order to clarify the indications for splenectomy in patients with homozygous β thalassaemia we studied, retrospectively, the basal pre‐transfusion haemoglobin levels and blood transfusion requirements before and after splenectomy, in a series of patients with this disorder.

Thirty‐six patients, of whom 20 underwent splenectomy, were included in this study. Three groups of patients with homozygous β thalassaemia were identified on the basis of clinical and laboratory findings. Two of the three groups consisted of 24 patients with β thalassaemia major, subdivided retrospectively according to their response to splenectomy. The third group consisted of six patients with β thalassaemia intermedia who had a comparably mild clinical course.

In the first group of 16 patients with β thalassaemia major, monthly transfusions were required from the age of 6 months to 24 months and splenectomy resulted in temporary improvement for a period of 1–2 years. Thereafter, the transfusion requirements returned to pre‐operative levels with no sustained improvement in base‐line pretransfusion haemoglobin levels. Despite the fact that haematological improvement was temporary, it is felt that splenectomy is indicated in this group since the development and the general well‐being of the children was significantly improved.

In the second group of eight patients with β thalassaemia major, transfusion requirements were relatively infrequent before 1–2 years, but after several years the transfusion requirement increased simultaneoulsy with the development of‘hypersplenism’. In these cases splenectomy was found to be beneficial for longer periods of time as manifested by a decrease in the transfusion requirement and an increase in the pretransfusion haemoglobin levels. Patients who developed hypersplenism relatively late in the course of their disease were usually the ones who derived long‐term benefit from splenectomy.

In the third group of six patients with β thalassaemia intermedia and a benign clinical course, with higher basal haemoglobin levels and lower blood transfusion requirements, most of the splenectomies were performed because of mechanical pressure due to splenomegaly. After splenectomy, these patients showed continued improvement in their haemoglobin levels and decreased transfusion requirements.
Partial Characterization of the Factor VIII‐Stimulating Material of Rabbit Liver Perfusates: Relationships to Rabbit Plasma Factor VIII
Authors: Benson R. E. , Jean Dodds W.
Published: March 2008

Summary.

Previous studies in our laboratory showed that effluent from perfused rabbit livers stimulates the production of factor‐VIII activity in perfused rabbit spleens. When concentrated supernatants of the hepatic perfusates were chromatographed on Sephadex G‐200, BioGel A‐15M and Sepharose 4B, the fractions with the greatest factor VIII‐stimulating capacity eluted in the void volume (V°). Sodium dodecyl sulphate (SDS) polyacrylamide gel electrophoresis of the Sephadex G‐200 V° fraction revealed components with a wide range of molecular weights. Rabbit factor VIII was isolated from ethanol‐concentrated, citrated rabbit plasma by Sepharose 4B filtration. The factor‐VIII activity appeared in the V° when eluted with physiological saline but was retarded on columns eluted in 0.8 M sodium chloride. Further analysis with SDS gels demonstrated that the Sepharose‐purified factor‐VIII preparation entered the polyacrylamide gels only after reduction with 2‐mercaptoethanol. Goat antirabbit factor VIII antiserum prepared against the gel‐purified antigen contained factor VIII‐neutralizing activity, was apparently monospecific by double‐diffusion and produced faint, single peaks by electro‐immunodiffusion. Antibody neutralization studies with the antiserum showed that the concentrated supernatants of hepatic perfusates contain fourfold greater factor VIII‐neutralizing antigen than procoagulant activity, whereas control perfusates, lacking factor VIII‐stimulating capacity, had negligible antibody‐blocking effect. The presence in hepatic perfusates of both high‐molecular‐weight factor VIII‐stimulating material and large amounts of factor VIII‐neutralizing antigen lacking procoagulant activity suggests that they are interrelated.
Microthrombocytosis and Platelet Fragmentation Associated with Idiopathic/Autoimmune Thrombocytopenic Purpura
Authors: Khan Iqbal , Zucker‐Franklin Dorothea , Karpatkin Simon
Published: March 2008

Summary.

Platelet volume distribution curves were obtained in 20 control subjects and in 21 patients with idiopathic/autoimmune thrombocytopenic purpura. A striking increase in microthrombocytes as well as megathrombocytes was noted in 86% of patients on one or more occasions, particularly in the presence of severe thrombocytopenia. The entire spectrum of platelet volume distribution curves noted in patients could be reproduced experimentally in rabbits following intravenous injection of anti‐platelet antibody. Differential centrifugation studies with control subjects revealed that microthrombocytes were light platelets and megathrombocytes were heavy platelets. Electron microscopy in patients with thrombocytopenia revealed that microthrombocytes were composed of intact small platelets as well as platelet fragments. It is concluded that severe peripheral destruction of platelets is associated with an increase in microthrombocytes as well as megathrombocytes.
Interactions between Genetic Red‐Cell Defects and the Environment
Authors: Luzzatto Lucio
Published: March 2008

Summary.

Environmental factors can affect the course of genetically determined disorders of red cells. On the other hand, genetically‘abnormal’red cells can respond to environmental agents differently from normal red cells. Some of the best characterized interactions of this sort, e.g. the haemoglobin S system and the G6PD system, are briefly reviewed. Additional established or postulated interactions are mentioned, and some suggestions are made as to how pertinent evidence could be obtained and exploited with a view to optimizing the interactions between genetic characters of red cells and the environment.
The Cellular Basis of Autoimmunity
Authors: Playfair J. H. L.
Published: March 2008

Summary.

The cellular basis of normal antibody formation is reviewed, and the stages are indicated at which the process may be interrupted in order to prevent the formation of autoantibodies. Evidence is presented for several types of interruption, including clonal elimination of lymphocytes, blocking of receptors, active suppression, and target blocking.
Autoimmunity to DNA
Authors: Fournié Gilbert J. , Lambert Paul H.
Published: March 2008

Summary.

The development of an immune response to DNA appears to be under a genetic control similar to that regulating the immune response to complex antigens. The predisposition of high responders is particularly marked at the B cell level, while a major role of suppressor T cells is not evident in this response. Various triggering factors may induce the formation of anti‐DNA antibodies: viral, bacterial or parasitic infections; tissue destruction or some drugs. A mechanism which may play an important role is the‘non‐specific’triggering of anti‐DNA antibodies by agents or substances which, like bacterial lipopolysaccharides, exert a potent mito‐genic effect on B cells and simultaneously induce a release of DNA in extracellular fluids.

Pathogenic effects of anti‐DNA antibodies are mainly due to the formation of DNA‐anti‐DNA complexes and to their subsequent localization in tissues.
Bone Marrow Transplantation for Aplastic Anaemia
Authors: Storb Rainer , Thomas E. Donnall
Published: March 2008

Summary.

Indications for treatment of aplastic anaemia (AA) by marrow transplantation are discussed. Selection of donor‐recipient pairs by histocompatibility testing, conditioning of patients for transplantation, technique of grafting, post‐grafting care and immunosuppressive drug regimens are reviewed. Data are presented on 28 patients with advanced AA treated by marrow grafts from HL‐A matched siblings following conditioning with cyclophosphamide (50 mg/kg) on each of 4 successive days. Survival and haematological reconstitution in more than half the patients indicates that marrow grafting in patients with advanced AA should be undertaken early, before major infections and refractoriness to blood transfusions occur provided that an HL‐A matched sibling can be identified as a donor. The results have shown that survival in marrow grafted patients with severe AA compares favourably to that of patients treated with conventional therapy.
Immunotherapy in the Management of Leukaemia
Authors: Fairley G. Hamilton
Published: March 2008

Summary.

Now that it has been clearly established that tumour associated antigens exist in acute leukaemia in man, as in animals, the possibility of stimulating the patient's immune system to react against them arises. In animal experiments the most effective method of influencing the progress of leukaemias and lymphomas after the implantation of living malignant cells has been a combination of nonspecific stimulation of the reticulo‐endothelial system, with agents such as BCG or Corynebacterium parvum, either with chemotherapy or with specific immunization with irradiated leukaemic cells. However, such treatment is only effective if the number of living malignant cells is small as it takes a powerful immune response to overcome even a small number of malignant cells. It is for these reasons that most of the studies in man have been on patients with acute leukaemia in remission. Mathé, in 1969, produced evidence that BCG and irradiated allogeneic leukaemia cells could lengthen the duration of remission in ALL in children, but the subsequent results of intensive combination chemotherapy together with prophylactic treatment of the central nervous system by Pinkel and his colleagues are so encouraging that immunotherapy is not being extensively used in this form of leukaemia at the moment. The situation in AML, particularly in adults, is completely different. The maintenance of remission with chemotherapy in this type of leukaemia is difficult and relapses occur quite rapidly. Various centres have now shown that both remission lengths and overall survival are significantly prolonged by using BCG with or without irradiated allogeneic leukaemia cells. The significance of these are reviewed and the problems for the future are discussed.
The Use of an Image Analysing Computer for the Quantitation of Red Cell Morphological Characteristics
Authors: Bentley S. A. , Lewis S. M.
Published: March 2008

Summary. Measurements of integrated optical density and projected area of red cells have been made on Romanowsky‐stained blood films from 25 subjects using an image analysing computer. Close correlation was observed between mean cell projected area and MCV and between mean cell integrated optical density and MCH. Mean values for the quotient of cell integrated density/cell projected area (mean cell colour density) correlated with MCHC. Variances of cell area and colour density correlated subjectively with anisocytosis and anisochromasia, and histograms of these parameters demonstrated dual populations. The clinical application of these findings is discussed.
A Serum Haemagglutinating Property Dependent upon Polycarboxyl Groups
Authors: Beck M. L. , Freihaut B. , Henry R. , Pierce S. , Bayer W. L.
Published: March 2008

Summary. A serum agglutinin reactive with red cells in the presence of polycarboxyl groups is reported. It is likely that this represents an additional example of the type of agglutinin previously described as agglutinating red cells in the absence of ionized calcium.

Experimental evidence is presented indicating that it is free polycarboxyl groups that potentiate agglutination and that any metal ion, such as calcium, capable of chelating with these groups will prove to be inhibitory.
Four New Pyruvate Kinase (PK) Variants and a Classical PK Deficiency
Authors: Miwa S. , Nakashima K , Ariyoshi K. , Shinohara K. , Oda E. , Tanaka T.
Published: March 2008

Summary. Four new red‐cell pyruvate kinase (PK) variants are presented along with one case of so‐called classical type PK deficiency. PK ‘Tokyo II’ had a low activity, Km (PEP) and Vmax, but a normal urea stability and only slight deviation from normal in neutralization tests by antiserum. It had a normal nucleotide specificity, abnormal electrophoretic mobility (fast moving) and the variant was associated with a mild haemolytic anaemia. PK ‘Maebashi’ had a low activity, high Km (PEP), low Vmax urea instability, decreased reactivity to antiserum, normal electrophoretic mobility, normal nucleotide specificity and was associated with a moderate haemolytic anaemia. PK ‘Tsukiji’ had low activity, high Km (PEP), markedly high Vmax, urea instability, decreased reactivity to antiserum, abnormal electrophoretic mobility (fast moving) and grossly abnormal nucleotide specificity especially abnormal behaviour to ADP. The haemolytic process in this case was moderate to severe. PK ‘Ube’ was electrophoretically abnormal (fast moving) but otherwise had normal characteristics and the propositus was healthy and not anaemic. PK ‘Ube’ was found by electrophoretic screening for genetic PK polymorphism. In the classical type PK deficiency, the usual red‐cell PK (PK‐R1 and PK‐R2) was not demonstrable by electrophoresis but instead M2‐type PK was present, presumably by compensatory process. Kinetic studies confirmed that the patient's red‐cell PK consisted of M2‐type PK. This patient had a severe haemolytic anaemia.
The Localization of Factor V within Normal Human Platelets and the Demonstration of a Platelet‐Factor V Antigen in Congenital Factor V Deficiency
Authors: Breederveld K. , Giddings J. C. , ten Cate J. W. , Bloom A. L.
Published: March 2008

Summary. Separation of human platelets from plasma by a modified gel‐filtration technique reveals very low levels of factor‐V activity of the platelet suspension. Repeated freezing and thawing increases the factor‐V activity in various factor‐V assays. This activity neutralized the inactivating effect of a rabbit‐antihuman factor V antibody to plasma factor V, while intact platelets had almost no such capacity. Washed and normal platelets and gel filtered platelets showed marked positive fluorescence after treatment with antifactor V serum and FITC labelled sheep anti‐rabbit immunoglobulin. Fluorescence was inhibited by previous incubation of the antifactor V serum and platelet lysates. Platelets of a factor V deficient patient showed the same fluorescence pattern as normal platelets indicating that they contained a factor V antigen. These platelets showed after lysis no effect in various factor V assays. From these studies it is concluded that the localization of factor V is within the platelets.
Continuous Flow Centrifuge Leucapheresis in the Management of Chronic Myelogenous Leukaemia
Authors: Hadlock D. C. , Fortuny I. E. , McCullough J. , Kennedy B. J.
Published: March 2008

Summary. Six patients with chronic myelogenous leukaemia (CML) were treated by repeated continuous flow centrifuge (CFC) leucapheresis. In six of seven courses of leucapheresis there was no improvement in splenomegaly. In no instance was the leucocyte count reduced to the normal range. The immediate changes in peripheral leucocytosis produced by each CFC leucapheresis procedure were quite variable. There was no correlation between the intensity of a series of CFC leucapheresis procedures and the total clinical effectiveness of the therapy. Balancing potential benefits against definite costs to the patient, CFC leucapheresis seems justified in only a minority of CML patients. Untreated CML patients who have cyclic leucocytosis may be the best candidates for this therapy. Currently this form of therapy for CML remains experimental.
Differences between Human Fe1‐Transferrin Molecules
Authors: Lane R. S.
Published: March 2008

Summary. TWO molecular forms of Fe1‐transferrin molecule can be demonstrated by anion‐exchange chromatography on columns of DEAE‐cellulose, pH 7.90. It is believed that co‐ordination of each metal‐binding site by ferric ions induces dissimilar changes in molecular conformation in the region of the binding sites which, in the case of one site, results in the molecule behaving as a weaker anion during anion‐exchange chromatography. If so, this could be entirely in accord with current views that the two binding sites of transferrin do not share equal properties of iron exchange with cells. At physiological levels of transferrin iron‐saturation, it is likely that two populations of Fe!‐transferrin molecules form the bulk of the iron‐transferrin complex.