bluuebunny/crossref_metadata_2025 huggingface dataset:
The role of haem biosynthetic and degradative enzymes in erythroid colony development: the effect of haemin
Authors: Ibrahim Nader G. , Lutton John D. , Levere Richard D.
Published: March 2008

Summary. In vitro culture of murine bone marrow has proved to be a useful system for defining the haem biosynthetic and degradative enzymatic pathways during erythroid colony (CFU‐E) growth and development. Previous attempts to elucidate these pathways during erythropoiesis have been limited by the amount of biological material available as well as the sensitivity of specific enzyme assays. These conditions were overcome in the present study. In order to obtain an enrichment of CFU‐E, nonadherent mouse bone‐marrow cells were cultured in special culture plates using a modified methyl‐cellulose medium with erythropoietin. CFU‐E yields were increased approximately threefold. Utilizing the sensitive radiochemical assay, direct measurement of ALAS activity was determined in very early developing CFU‐E cultures as well as mature cultures. ALAS activity was found to reach a peak after 60 h of culture growth and then started to decline in activity. Cellular synthesis of haem was determined (with 14C ALA) and minor modifications of standard assays were also made in order to determine spectrophotometrically δ‐aminolaevulinic acid dehydratase (ALAD) and haem oxygenase activity in developing CFU‐E cultures. These assays were reproducible with as few as four or five culture plates. It was found that ALAD activity rose progressively after 36 h of culture growth and reached a plateau at about 60 h of growth. A continuous increase in 14C ALA incorporation into haem was seen at later hours of culture growth, suggesting that the induction of other haem enzymes beyond ALAS is still rate limiting in haem synthesis. Conversely, haem oxygenase activity declined up to 60 h of growth and was elevated at later culture periods without a subsequent increase in ALASA and ALAD. The observed increase in haem enzymes brought about by haemin was completely suppressed by addition of cycloheximide to the cultures. These results suggest that
Transient dyserythropoiesis in repopulated human bone marrow following transplantation: an ultrastructural study
Authors: Rozman C. , Feliu E. , Granẽna A. , Brugues R. , Woessner S. , Corrons J. L. Vives
Published: March 2008

Summary. Transmission electron microscopy was used to examine marrow samples from 15 patients with aplastic anaemia or acute leukaemia who had been treated with bone marrow transplantation. There were 11 allogeneic, three syngeneic and one autologous graft. The purpose was to estimate the frequency, type and extent of dyserythropoietic change. Transient dyserythropoietic features were substantiated in all cases. Nuclear changes were present in 12 cases, iron laden mitochondria (sideroachrestic phenomena) in 10 and cytoplasmic contacts and/or connections between red cell precursors in 10. Dyserythropoiesis was most conspicuous in the majority of cases between 14 and 28 d after transplantation but it may persist for over 100 d. No deficit in red cell production was noted and it is proposed that dyserythropoiesis in this circumstance is a physiological rather than a pathological phenomenon.
Immuno‐electron microscopic tracing of lactoferrin, a regulator of myelopoiesis, into a subpopulation of human peripheral blood monocytes
Authors: Steinmann G. , Broxmeyer H. E. , de Harven E. , Moore M. A. S.
Published: March 2008

Summary. Lactoferrin (LF), an iron binding glycoprotein present in the specific granules of mature granulocytes, is capable of suppressing the production of granulocyte‐macrophage colony stimulatory factors (GM‐CSF) by monocytes and macrophages in vitro and rebound myelopoiesis in vivo. By an immuno‐electron microscopic technique (peroxidase‐antiperoxidase procedure) in combination with a three‐step fixation and detergent treatment of mononuclear blood cells pulsed with LF, intracellular localization of LF in a sub‐population of human monocytes with a well‐preserved subcellular morphology was demonstrated. LF was found in the monocytes after a 30 min pulse with concentrations of LF as low as 10−14m. Under the electron microscope, deposits of approximately 110 nm diameter appeared scattered in the area of the endoplasmic reticulum. Large numbers of smaller deposits (20–70 nm) were found in the euchromatin. The heterochromatin and other cytoplasmic organelles were free of LF. These results suggest that the euchromatin might be the functional site for LF inhibition of the production of GM‐CSF by a subpopulation of human monocytes.
Unusual intracytoplasmic immunoglobulin inclusions in chronic lymphocytic leukaemia
Authors: Guglielmi P. , Preud'homme J. L. , Gourdin Marie‐Françoise , Reyes F. , Daniel Marie‐Thérèse
Published: March 2008

Summary. Unusual intracytoplasmic immunoglobulin inclusions were found by immunofluorescence in three patients with chronic lymphocytic leukaemia. The inclusions contained the same immunoglobulin chains as those detected on the plasma membrane, except for δ chains which were expressed on the cell surface and not in the cytoplasmic inclusions. The cytoplasmic staining persisted throughout culture for 8 or more days. An initial study of patients l's cells showed that the inclusions contained only μ chains, and κ chains gradually became apparent after in vitro culture. In a second study, the fresh lymphocytes contained both μ and κ chains. Initially, biosynthetic experiments showed production of μ chains which polymerized in the cytoplasm and were not secreted. Subsequently there was synthesis of heavy and light chains which assembled into monomeric subunits that were retained and secretion of free light chains. The apparent molecular weight of these immunoglobulin chains was larger than that of their secretory counterparts. Immunoelectronmicroscopy revealed cytoplasmic μ chains in strands of endoplasmic reticulum. In the two other patients, immunofluorescence displayed unusual staining patterns of bright networks in perinuclear areas.
Effect of ADP‐utilizing enzymes on the arterial bleeding time in rats and rabbits
Authors: Zawilska Krystyna M. , Born G. V. R. , Begent Nicola A.
Published: March 2008

SUMMARY. Platelets are activated in vitro by various chemical agents. It is not certain which if any of these is responsible for the haemostatic aggregation of platelets in living blood vessels. This report provides evidence that ADP is involved in activating platelets in vivo.
A novel semi‐synthetic sulphated polysaccharide with potent antithrombin activity
Authors: Nilsson I. M. , Rothman U. , Stenberg P. , Frohm B. , Persson N. H.
Published: March 2008

A mucopolysaccharide was isolated from mussel broth. After sulphation, an electrophoretically homogenous product was obtained with a molecular weight of approximately 40 000 daltons and a sulphur content of about 12%. The sulphated polysaccharide (S‐Lim) displayed an anticoagulant activity in a thrombin test system with human plasma. Unlike heparin, the anticoagulant effect of S‐Lim was observed also in a thrombin‐fibrinogen clotting system in the absence of AT III. Complete inhibition of the effect of 1·0 mg S‐Lim was achieved with 1·5 mg protamine. In an activated partial thromboplastin time test system the anticoagulant activity of 1.0 mg S‐Lim corresponded to about 40 iu of heparin. S‐Lim was also found to inhibit thrombin‐induced platelet aggregation. Furthermore, S‐Lim inhibited the thrombin‐dependent activation of factor XIII in human plasma. S‐Lim did not affect various test systems measuring factor Xa activity. It is concluded that this new sulphated mucopolysaccharide acts as a pure antithrombin with a potency corresponding to 40‐50 iu heparin/mg S‐Lim. In contrast to heparin and other heparinoids it does not require the presence of AT III.
Greek (Aγ) variant of hereditary persistence of fetal haemoglobin: globin gene organization and studies of expression of fetal haemoglobins in clonal erythroid cultures
Authors: Papayannopoulou Th. , Lawn R. M. , Stamatoyannopoulos G. , Maniatis T.
Published: March 2008

Individuals heterozygous for the Greek (Aγ) variant of hereditary persistence of fetal haemoglobin (HPFH) synthesize Hb F whose γ‐globin chains are predominantly of the Aγ type. DNA obtained from Greek HPFH heterozygotes was used to test for abnormalities in the organization of non α‐globin genes. In addition, γ‐ and β‐globin expression was studied in BFUe cultures. Restriction endonuclease mapping showed that the γGγ, δ and β genes in cis to the Greek HPFH determinant are intact. Overproduction of γ‐globin chains synthesis was observed in the BFUe cultures. A significant portion of the γ chain synthesis was of the Gγ type, suggesting that the Gγ genes cis and trans to the HPFH chromosome are active in culture.
Size dependent platelet subpopulations: relationship of platelet volume to ultrastructure, enzymatic activity, and function
Authors: Thompson Craig B. , Eaton Katherine A. , Princiotta Steven M. , Rushin Carol A. , Valeri C. Robert
Published: March 2008

A method for the separation of platelets on the basis of their size has been developed using counterflow centrifugation. Platelets were separated, free of plasma proteins and other cells, into seven subpopulations. The smallest‐sized platelets, designated as Fraction 1, had a mean platelet volume (MPV) of 3.94 ± 0.60 μm3 (SD). Each successive fraction had a progressively larger MPV. The MPV for the largest‐sized platelets, designated Fraction 7, was 8.19 ± 0.64 μm3. The MPV for the original platelets prior to fractionation was 6.57 ± 0.61 μm3. The mean density of Fraction 1 platelets was 1.067 ± 0.002 g/cm3, while Fraction 7 had a mean density of 1.072 ± 0.001 g/cm3. Transmission electron microscopy demonstrated that Fraction 1 had 4.3 ± 0.9 dense bodies per platelet, and Fraction 7 had 12.6 ± 2.4 dense bodies per platelet. Platelet LDH activity showed that the Fraction 1 platelets had 4.77 ± 0.92 iu per 1010 platelets; Fraction 7 platelets had 14.88 ± 1.23 iu per 1010 platelets. The LDH activity in the platelets before separation into subpopulations was 9.47 ± 1.45 iu per 1010 platelets.
An acquired coagulation inhibitor to factor II
Authors: Scully M. F. , Ellis V. , Kakkar V. V. , Savidge G. F. , Williams Y. F. , Sterndale H.
Published: March 2008

An elderly woman with a bleeding problem associated with prolonged PTT, PT and thrombin time but with normal snake venom time was found to have very high levels of plasma antithrombin. The antithrombin activity was isolated and characterized and found to be an IgG which directly inhibited thrombin and also precipitated prothrombin. The concentration in plasma of the γ‐globulin inhibitory activity was equivalent per ml to 80 NIH units of thrombin. Corticosteroid and immunosuppressive treatment reduced the level of the inhibitor with remission of bleeding symptoms.
Busulphan aplasia in rabbits: a model for human aplastic anaemia
Authors: den Ottolander Gerard J. , te Velde Jan , Veenhof Wil , Kleiverda Karin , Haak Hans L. , Spaander Piet Jan
Published: March 2008

Summary. Bone marrow histology plays a crucial role in the clinical diagnosis of aplastic anaemia. The nature of the disease means that few studies are available on the histological changes which occur in the early stages of the development of aplasia. We describe here an animal model which may have some relevance in this respect. Rabbits were chronically exposed to busulphan (BU) to induce aplasia. Sequential histological monitoring of the bone marrow was performed to obtain information about the events preceding full‐blown aplasia. There was an early decrease and ultimate disappearance of granulo‐ and megakaryopoiesis with relative sparing of erythropoiesis which showed severe dysplasia. Increasing lymphoplasma‐cytoid infiltrate resembling that seen in human aplasia could be observed in the majority of the animals, together with a decrease of the peripheral lymphocyte number. Lymph nodes and spleen did not show lymphocyte depletion and serum gamma‐globulin remained stable. Fibrosis was observed in 50%, which is in contrast with human aplasia at diagnosis. In half of the animals there was a rise in MCV, which was not correlated with reticulocytosis or degree of dyserythropoiesis.

BU‐induced aplasia in rabbits, which resembles long‐standing grade II human aplasia in many respects, might be a suitable model for the study of aplastic anaemia due to stem cell defects.
Assessment of bone marrow histology in the malignant lymphomas (non‐Hodgkin's): correlation with clinical factors for diagnosis, prognosis, classification and staging
Authors: Bartl R. , Frisch B. , Burkhardt R. , Kettner G. , Mahl G. , Fateh‐Moghadam A. , Sund M.
Published: March 2008

Summary.

Bone marrow biopsies of 678 untreated patients with established malignant non‐Hodgkin's lymphomas (ML) were investigated. The bone marrow was involved in 468 cases, an overall frequency of 69%. The Kiel classification of the ML (based on lymph node histology) was applied and the biopsies were classified: ML lymphocytic 36%, ML ‘hairy cell’24%, ML lymphoplasmacytic/cytoid 24%, ML centrocytic 6%, ML centroblastic/centrocytic 4%, ML lymphoblastic (without ALL) 3%, ML centroblastic 2% and ML immunoblastic 1%. The life tables of the patients were similar whether classified according to the histology of the lymph node or the bone marrow. A multivariate computer based analysis of both clinical and histological data was performed to test their prognostic relevance. The cell type, the proliferation pattern and the extent of infiltration in the bone marrow all proved to be factors of prognostic significance. The results indicate that classification of the ML based on lymph node histology is applicable to the bone marrow, is reproducible and has prognostic significance. Consequently, a bone marrow biopsy is a useful clinical tool for staging and for histological classification of patients with ML.
Studies on quinine‐ and quinidine‐dependent antibodies against platelets and their reaction with platelets in the Bernard‐Soulier syndrome
Authors: van Leeuwen E. F. , Engelfriet C. P. , Kr. vondem Borne A. E. G.
Published: March 2008

Summary.

The sera of 14 patients with quinine/quinidine‐dependent thrombocytopenia were studied in the platelet suspension immunofluorescence test (PSIFT), the 51Cr‐lysis assay and the complement fixation test (CFT).

When anti‐Ig or anti‐IgG reagents were used, the PSIFT proved to be a little more sensitive than the 51Cr‐lysis assay and far more sensitive than the CFT. With the PSIFT, it was possible to determine the immunoglobulin class of the antibodies, which in all sera was IgG of the subclass IgGl. In two sera there were also IgG3 antibodies, and in the sera of three patients additional IgM drug‐dependent platelet antibodies were detected.

With the anti‐C3 reagent, however, lower titres were observed in the PSIFT than in the 51Cr‐lysis assay, in contrast to the results with other complement‐binding IgG platelet antibodies. The F(ab′), fragments of the antibodies did not react in the PSIFT. This indicates that the Fc part of the antibody is essential for the fixation on the platelet membrane. These findings support the theory that in this syndrome drug‐antibody complexes are formed in the absence of platelets that selectively adhere to the Fc receptor on platelets.

The platelets of patients with the Bernard‐Soulier syndrome (BSS) reacted normally with the quinine/quinidine‐dependent antibodies in some of the sera, but not with those in other sera in the PSIFT. However, lysis of BSS platelets was never observed. The results of absorption studies showed a reflection of the results in the PSIFT, indicating that the receptor for the quinine/quinidine‐dependent antibodies is not absent in this disease but altered. Whether a reaction with BSS platelets will take place depends on characteristics of the antibodies.
Changes in the life expectancy of patients with severe haemophilia A in Finland in 1930‐79
Authors: Ikkala E. , Helske T. , Myllylä G. , Nevanlinna H. R. , And P. Pitkänen , Rasi V.
Published: March 2008

Summary. Important advances have been made in the treatment of haemophilia during the past 30 years. We have analysed the data of all the known 163 patients with severe haemophilia A living in Finland in 1930–79 in order to study changes in the prognosis of severe haemophilia A. During the period of 50 years the mean age at death of the patients has increased from 7.8 years in 1930‐39 to 25.5 years in 1970–79 and the annual death rate has markedly decreased in all age groups. The decline has been greatest in patients under 10 years of age. In this age group the annual death rate decreased from over 50 per thousand in 1930‐39 and 1940‐49 to 4.8 per thousand in 1970–79. The prognosis of patients with inhibitors has remained poor, however. Five of the six deaths during the last decade occurred in patients with inhibitors. The overall annual death rate of patients without inhibitors was only 1.2 per thousand in 1970–79, suggesting that at the present time the life expectancy of patients who do not develop inhibitors does not markedly differ from that of the general male population.
Reversal of severe bone marrow fibrosis and osteosclerosis following allogeneic bone marrow transplantation for chronic granulocytic leukaemia
Authors: McGlave P. B. , Brunning R. D. , Hurd D. D. , Kim T. H.
Published: March 2008

Summary. Severe marrow fibrosis and osteosclerosis gradually disappeared after a 33‐year‐old woman received an allogeneic bone marrow transplantation (BMT) as experimental treatment for chronic granulocytic leukaemia. Serial biopsies demonstrate gradual resolution of dense reticulin fibrosis, collagen fibrosis and osteosclerosis, and restoration of normal marrow architecture after transplantation. These changes correspond with histological and cytogenetic evidence of normal marrow engraftment and sustained complete remission from chronic granulocytic leukaemia. In this case severe marrow infiltration with reticulin and collagen fibrosis as well as severe derangement of marrow architecture and obliteration of the medullary cavity by osteosclerosis was an entirely reversible process after allogeneic bone marrow transplantation, and did not prevent successful engraftment, haemopoietic and cytogenetic reconstitution and complete remission from chronic granulocytic leukaemia.
The 5q‐ chromosome abnormality in haematological disorders:|a collaborative study of 34 cases from the Netheralands
Authors: Kerkhofs H. , Hagemeijer A. , Leeksma C. H. W. , Abels J. , Den Ottolaner G. J. , Somers R. , Gerrilts W. B. J. , Lqangenhuiyuen M. M. A. C. , Van Dem Borne A. E. G. Kr. , Van Hemel J. O. , Geraedts J. P. M.
Published: March 2008

Summary. Clinical, haematological and cytogenetic data of all patients with an acquired 5q ‐ abnormality, diagnosed in four cytogenetic laboratories, were studied. The 5q‐ arose de novo in 24 patients, while 10 patients exhibited it after radiation and/or chemotherapy. Sixteen cases with a dysmyelopoietic syndrome (DMPS) and one case with acute nonlymphocytic leukaemia (ANLL) had no chromosome abnormalities other than 5q‐ . All nine patients with refractory anaemia (RA) belonging to the de novo group had a normal cell line. So far none of them has transformed to acute leukaemia, while three patients with refractory anaemia with excess of blasts (RAEB), pure red cell aplasia (PRCA) and acquired idiopathic sideroblastic anaemia (AISA), having the 5q‐ in 100% of the cells showed such a transformation. The second group consisted of nine patients with a 5q‐ and additional chromosome abnormalities: three of them died of progression of the disease, while four died of unrelated causes. Eight out of nine patients with a 5q‐ and acute leukaemia, either de novo (five) or secondary (four) to radiation and/or chemotherapy, had additional chromosome abnormalities. Among the changes (partial) monosomy of chromosome 7 was most frequently found.

The median survival time of the group of patients with an isolated 5q‐ in the presence of a normal cell line was significantly longer than that of patients with additional chromosomal changes with or without acute leukaemia.

All patients with an isolated 5q‐ abnormality showed the previously reported megakaryocytic abnormalities (small mono‐ or bilobulated). In most of the other patients these megakaryocytic abnormalities were also found, whereas others were definitely normal in this respect. A striking and unexplained preponderance of females (1 2 out of 16) was found in the group with an isolated 5q‐ anomaly.

All deletions studied with banding techniques appeared to be interstitial but the breakpoints were variable. The shortest region of overlap is near band 5q22.
Functional compartmentation of DNA precursors in human leukaemoblastoid cell lines
Authors: Taheri M. R. , Wickremasinghe R. G. , Hoffbrand A. V.
Published: March 2008

Summary. Human leukaemoblastoid cell lines were incubated with [3H]t hymidine and used to study the utilization for DNA synthesis and degradation of thymine nucleotides labelled via the salvage pathway. Three types of experiments were performed.

(1) Cells were continuously labelled with exogenous [3H]thymidine at 3 7°C to compare the incorporation of label into DNA with the specific radioactivities of thymine nucleotides separated by chromatography on polyethyleneimine‐cellulose. These studies demonstrated whether one or more functional pool of thymine nucleotides was present.

(2) Intracellular thymine nucleotide pools were labelled with [3H]thymidine at 13°C, and the cells were subsequently incubated in unlabelled medium at 37°C in order to quantify the percentage of thymine nucleotides incorporated into DNA. The results showed that 18%, 75%, 75% and 91% of intracellular thymine nucleotides were incorporated into DNA during the chase, in Raji (B lymphoid), Nalm‐6 (pre‐B cell), HL60 (myeloid) and KM3 (c‐ALL) cells respectively, whereas Molt4 (Thy‐ALL) cells incorporated 100%.

(3) In order to quantify the activity of the degradative pathway for thymine nucleotides in these cell lines, cells were labelled as described in (2) in the presence of cytosine arabinoside (an inhibitor of DNA synthesis). 100%, 90%, 65%, 38% and 20% of labelled thymine nucleotides were degraded in Raji, HL60, Nalm‐6, KM3 and Molt‐4 respectively.

These results confirm our previous conclusion (Taheri et al, 1981a, b) that thymine nucleotides are functionally compartmentalized in human cells. If these results with cell lines can be extrapolated to apply to de novo leukaemia cases, they suggest that the degree of compartmentation differs widely between different leukaemia cell types. Thy‐ALL cells utilized all their labelled thymine nucleotides for DNA synthesis, whereas B‐ALL, cells utilized only 18% of their labelled thymine nucleotide nucleotides. Thy‐ALL cells have the smallest degradative thymine pool and degradative capacity whereas B‐ALL cells possess the largest degradative pool and capacity. AML, pre‐B‐ALL, c‐ALL cell lines show intermediate results. If these results reflect those of the normal bone marrow cells from which each of the leukaemias arise, they suggest that with progressive B cell development in the bone marrow, degradative ability for DNA precursors increases and the efficiency of utilization of DNA precursors for DNA synthesis decreases.
Plasma levels of immunoreactive erythropoietin after acute blood loss in man
Authors: Miller Marilyn E. , Cronkite Eugene P. , Garcia Joseph F.
Published: March 2008

Summary. Plasma levels of immunoreactive erythropoietin (Ep) were measured after acute blood loss (50,75,200 and 450 ml) in man in order to determine the volume of blood loss required to trigger an Ep response as well as to define the reticulocyte response. There was a highly significant (P <0.01) linear increase in reticulocyte count after 200 and 450 ml of blood loss. Analysis of trends also showed a highly significant (P < 0.01) linear response of haematocrit and Ep after a 450 ml blood loss. The reticulocyte count increases were not dependent on a prior increase in Ep level indicating that at least two mechanisms are operative in restoring the red cell mass to normal after blood loss.
Quantitative histology of myeloma‐induced bone changes
Authors: Valentin‐Opran Alexandre , Charhon Sam A. , Meunier Pierre J. , Edouard Claude M. , Arlot Monique E.
Published: March 2008

Summary. In order to quantify bone changes which occur in multiple myeloma, undecalcified transiliac bone biopsies from 118 myelomatous patients were analysed by histomorphometric methods. Osteoclastic resorption surfaces were increased compared with controls, and the number of osteoclasts/mm2 of bone section was significantly greater in the areas massively invaded by plasma cells than in less invaded areas. The osteoid surfaces were also increased and the percentage of trabeculae that exhibited tetracycline labelling was also greater, indicating increased formation surfaces. However, reduced thickness of the osteoid seams and a low calcification rate, measured after tetracycline double labelling, suggests a reduced activity for each osteoblast. The mean trabecular bone volume was not reduced as compared with controls, but the biopsies showed a heterogeneous distribution of osteolytic and osteosclerotic areas. In the invaded areas, no major histomorphometric difference was found between patients receiving chemotherapy and untreated patients, demonstrating that if usual chemotherapies reduce the tumour mass, they do not improve histological bone lesions in areas still invaded by plasma cells.
Philadelphia chromosome‐positive blastic leukaemia: ultrastructural and ultracytochemical evidence of basophil and mast cell differentiation
Authors: Parkin Janet L. , McKenna Robert W. , Brunning Richard D.
Published: March 2008

Summary. Ultrastructural, ultracytochemical, immunologic and biochemical studies were performed on leukaemic cells from 41 patients with Philadelphia chromosome‐positive blastic leukaemia; 28 patients were in blast transformation of chronic myelogenous leukaemia and 13 patients presented with ‘acute’leukaemia. The patients were divided into two morphologic groups, lymphoid (16 cases) and myeloid (25 cases), on the basis of light microscopy and cytochemistry. All lymphoid cases studied for the presence of CALLA (10 patients) and TdT (11 patients) were positive. Two of 13 myeloid cases studied were TdT positive. The blasts from 10 of 16 lymphoid cases contained immature basophil/mast cell granules on ultrastructural examination. Peroxidase‐positive ‘lymphoid’blasts were noted in three of seven patients studied by ultracytochemical techniques. The reactivity was primarily confined to granular structures. Of the 25 cases in the myeloid group, blasts from 14 cases showed basophil/mast cell differentiation, nine cases showed neutrophil/monocyte features, and two cases were megakaryoblastic. Distinct patterns of ultrastructural peroxidase positivity were seen in the seven myeloid cases studied. In basophil/mast cell precursors the reactivity was primarily confined to granules; neutrophil precursors showed reactivity in the nuclear envelope, rough endoplasmic reticulum (RER), golgi and granules; in megakaryoblasts, only the nuclear envelope and RER were positive while the granules were consistently negative.
Assessment of marrow trephine in relation to staging in chronic lymphocytic leukaemia
Authors: Bartl R. , Frisch B. , Burkhardt R. , Hoffmann‐Fezer G. , Demmler K. , Sund M.
Published: March 2008

The prognostic value of the Rai clinical staging system and of a series of bone marrow features was examined in 167 untreated patients with CLL. The patients fell into three prognostic groups: those with limited disease, Rai stages 0 and I, median survival 107 months; those with intermediate disease, Rai stage II, median survival 44 months; and those with extensive disease, Rai stages III and IV, median survival 27 months. Two types of infiltration patterns, diffuse and nodular, were found in the bone marrow, and these were associated with significantly different median survival times: 34 and 115 months respectively.

It is suggested that diffuse and nodular patterns in the bone marrow relate to the rapidity of progress of the disease rather than to different stages in CLL.

The data suggest that a simplified clinical staging system together with an histological classification reflects both the extent as well as the rate of progression of CLL. The course of CLL may be monitored by sequential bone marrow biopsies to assess changes in the proliferation pattern, in the proliferative cell system and in the tumour mass, which give early warning of shifts in the rate of progression and therefore in the prognosis of the disease.
Immune‐mediated platelet destruction and thrombocytopenia in patients with solid tumours
Authors: Schwartz Kenneth A. , Slichter Sherrill J. , Harker Laurence A.
Published: March 2008

Autoimmune mediated platelet destruction with severe thrombocytopenia was documented in eight patients with solid tumours. The patients had reduced platelet lifespan, positive platelet antibody tests, increased numbers of megakaryocytes, and a rise in platelet count following treatment with steroids, splenectomy or immunosuppressive therapy. Intravascular coagulation was excluded as the predominant cause of thrombocytopenia by near normal 125I‐fibrinogen survival; thrombocytopenia secondary to marrow suppression was ruled out by increased platelet turnover. Thus, like patients with lymphoproliferative disorders, patients with solid tumours may be thrombocytopenic because of immune mediated platelet destruction.
Simplified immunoradiometric assay for factor VIII coagulant antigen
Authors: Thomas K. B. , Howard M. A. , Koutts J. , Firkin B. G.
Published: March 2008

A simplified, non‐competitive, solid phase immunoradiometric assay has been developed for the quantitation of factor VIII coagulant antigen (VIII:CAg)—the antigenic counterpart of FVIII coagulant activity (VIII:C). Both homologous and heterologous antibodies to human factor VIII (FVIII) were used in this assay. Initially, FVIII in a test sample was attached to immobilized, human IgG obtained from a polytransfused haemophilia A patient with a high titre antibody to VIII:C. The bound FVIII was then detected using rabbit 125I‐IgG specific for human FVIII. The concentration of VIII:CAg correlated well with VIII:C levels in the plasma from normal donors (r= 0.84, n= 15).

Homozygote von Willebrand's disease patients had undetectable levels of VIII:CAg in their plasma. Patients with severe haemophilia A (VIII:C < 0.01 u/ml) could be divided into groups on the basis of the VIII:CAg levels, i.e. those having undetectable VIII:CAg and others with measurable VIII:CAg. VIII:CAg detected in normal serum was < 0.002 u/ml.

In this assay the use of human antibody to FVIII is considerably decreased compared to other methods for VIII:CAg, and the time‐consuming steps to immunopurify human anti‐FVIII antibody are eliminated.
Uptake and release of transferrin and iron by mitogen‐stimulated human lymphocytes
Authors: Bomford Adrian , Young Stephen P. , Nouri‐Aria Kayhan , Williams Roger
Published: March 2008

Summary.

Phytohaemagglutinin stimulation of human peripheral blood lymphocytes resulted in the expression of transferrin receptors and the uptake of iron into the cells. As assessed from the resistance of the 125I label to pronase, transferrin was rapidly bound and internalized at 37°C, while at 4°C 85% of the 125I label remained on the cell surface and was degraded by the pronase. Over 94% of the 125I label associated with and subsequently released from the cells was acid precipitable, indicating that transferrin was not degraded during its uptake and release. After reincubation for 1 h in fresh medium 70% of the cell associated 125I‐transferrin was released. In contrast, less than 30% of the 59Fe was released, showing that iron was removed from transferrin and retained by the cells.

Concentration dependent binding of 125I‐transferrin estimated at 37°C occurred with an apparent Ka of 5·7±1·1 × 107l mol−1 (mean ± SD, n= 4) indicating little variation between cells from different individuals, although the number of transferrin molecules associated with the cells varied greatly from 6·2 × 104/cell to 1·4 × 105/cell. The rate of iron uptake from 59Fe and 125I labelled transferrin at 37°C by the cells from different subjects was also very variable, with a range between 0·46 and 2·27 pg Fe/min/106 cells (n= 6). However, iron uptake did not correlate with the amount of transferrin bound. This suggests that transferrin uptake and the release of iron from the transferrin to the interior of the cell are controlled independently.
Detection of glycoprotein IIb and IIIa by monoclonal antibodies
Authors: Thurlow P. J. , Barlow B. , Connellan J. M. , McKenzie I. F. C.
Published: March 2008

Summary.

Murine monoclonal antibodies were produced against human platelet membranes and screened on platelets by a 125I protein A radioimmunoassay. Several clones produced platelet specific antibodies as they showed no reaction with peripheral blood lymphocytes, neutrophils, bone marrow (excluding megakaryocytes) or several cell lines. Two antibodies (designated anti‐HuPl‐mla and anti‐HuPl‐mlb) were of particular interest in that although platelet specific they were non‐reactive with platelets from a thrombasthenic patient. In functional assays these two antibodies could specifically inhibit ADP and collagen induced aggregation of platelets and release of ATP, retard platelet aggregation and ATP release induced by epinephrine, and inhibit ADP induced platelet fibrinogen binding. These two antibodies appear to recognize glycoproteins IIb and IIIa as analysis by SDS‐PAGE using radiolabelled membranes revealed a two chain structure of molecular weight 112 000 and 122 000 daltons when run after reduction and 87 000 and 140 000 daltons non‐reduced.
Mitogen‐induced stimulation and suppression of erythroid burst promoting activity production by human mononuclear cells
Authors: Zuckerman Kenneth S. , Haak Mary
Published: March 2008

Summary.

Exposure of human peripheral blood mononuclear cells or highly enriched monocytes to various plant lectins substantially alters their production of erythroid burst promoting activity (BPA). Neither unstimulated, nor mitogen stimulated, enriched T lymphocytes produced demonstrable BPA. Each of the lectins tested resulted in a different pattern of alteration of BPA production by mononuclear cells. Increasing concentrations of phytohaemagglutinin (PHA) caused a progressive increase in BPA production up to a plateau level at concentrations above 0·25–0·5 μ1/ml. Concanavalin A (Con A) at concentrations of 0·05–0·1 μg/ml stimulated BPA production, but Con A concentrations > 1 μg/ml never augmented BPA production by mononuclear cells. Pokeweed mitogen inhibited BPA production by mononuclear cells in a concentration‐dependent manner. Since PHA and Con A can bind to and stimulate both monocytes/macrophages and T lymphocytes, some production of BPA by stimulated T cells in the presence of monocytes cannot be ruled out. Earlier studies demonstrated that T cells augment monocyte production of BPA. Thus, monocyte–T cell interactions, as well as activation of monocytes and perhaps lymphocytes, play an important role in regulation of BPA production in vitro.
Plasma fibronectin in myeloproliferative disorders and chronic granulocytic leukaemia
Authors: Norfolk D. R. , Bowen M. , Roberts B. E. , Child J. A.
Published: March 2008

Summary.

A significant reduction of plasma fibronectin levels was found in polycythaemia vera and myelofibrosis, the lowest levels being found in patients with marked splenomegaly. Plasma fibronectin concentration was normal in essential thrombocythaemia, and only modest reduction was seen in chronic granulocytic leukaemia in either controlled chronic phase or blast cell crisis. In a patient with myelofibrosis, the plasma fibronectin rose from less than 100 mg/l to 177 mg/l after splenectomy. Possible explanations include increased consumption of plasma fibronectin in the expanded mononuclear phagocyte system present in the liver and spleen, reduced hepatic synthesis, and the clearance of circulating immune complexes. Low plasma fibronectin concentrations may increase susceptibility to infection.
The effects of surgery on the activity of neutrophil granule proteins
Authors: Davies J. M. , Sheppard K. , Fletcher J.
Published: March 2008

Summary. Activities of the neutrophil granule‐associated proteins β‐glucuronidase, lysozyme and vitamin B12 binding protein were measured, serially, in the cells and serum of 10 patients undergoing total abdominal hysterectomy.

The neutrophil leucocytosis which followed total abdominal hysterectomy was accompanied by a fall in the intraneutrophilic activities of all three granule‐associated proteins.

Intraneutrophilic lysozyme activity and intraneutrophilic vitamin B12 binding capacity were maximally reduced within 4 h of surgery and fell to 62 ± 13% (mean ± SEM) and 63 ± 9% of their preoperative levels, respectively. This contrasted with the activity of intraneutrophilic β‐glucuronidase which was not maximally reduced until 24 h post‐surgery when a fall to 80 ± 6% of the preoperative level was observed.

By the fifth postoperative day activities of the three intraneutrophilic granule proteins were increasing and approaching those observed preoperatively.

Serum lysozyme and plasma unsaturated vitamin B12 binding capacity (UBBC) rose steadily following surgery and were significantly elevated by the fifth postoperative day.

It is suggested that activation and in vivo degranulation of circulating neutrophils may be responsible for these changes in activity of neutrophil granule proteins following surgery.
Granulocyte function in untreated pernicious anaemia
Authors: Kätkä K. , Skger R. A. , Matsunaga T. , Toivanen Auli , Hitzig W. H.
Published: March 2008

Summary. Neutrophil granulocyte functions were evaluated in nine patients with pernicious anaemia (PA). Membrane potential, oxygen consumption, nitroblue tetrozolium reduction, and phagocytosis and killing of staphylococci were determined prior to the institution of vitamin B12 therapy. There were no significant alterations in these granulocyte functions as compared with normal controls, although in several patients low values were seen. We conclude that in PA the oxidative metabolism and microbicidal functions of granulocytes in general are normal, even in severely anaemic patients.
Idiopathic autoantibody that inhibits fibrin monomer polymerization
Authors: Ghosh S. , McEvoy P. , McVerry B. A.
Published: March 2008

Summary. A 73‐year‐old female was found to have prolonged thrombin and reptilase times in the immediate post‐operative period. These abnormalities were not corrected by the addition of normal plasma. They were subsequently shown to be due to an IgG immunoglobulin which inhibited fibrin monomer polymerization. The IgG immunoglobulin activity could be neutralized completely by prior incubation with either patient or normal fibrinogen, uncrosslinked fibrin monomers or IgG antisera. No inhibitory effect on thrombin activity, fibrinopeptide A release or on the fibrin cross‐linking reaction of factor XIIIa could be detected. Purified patient fibrinogen was functionally normal as demonstrated by normal fibrinogen–fibrin polymerization and fibrinopeptide A release. No underlying cause for this phenomenon was found. The presence of the inhibitor was associated with excessive blood loss during the post‐operative period.
Red cell nucleotide levels in patients with thalassaemia, iron deficiency, and renal failure
Authors: Crowther P. J. , Cauchi M. N.
Published: March 2008

Summary. Red cell nucleotides, predominantly adenine nucleotides, were measured spectrophotometrically in normal individuals and in patients with iron deficiency, β thalassaemia and renal failure following dialysis or transplantation. There was a significant correlation between the mean red cell haemoglobin (MCH) or the mean cell volume (MCV) and the nucleotide levels in red cells (P < 0·001). Patients with β thalassaemia did not have increased nucleotide levels compared to red cells having the same MCH or MCV. Patients with renal failure had significantly increased levels of nucleotides compared to normal individuals, and this level remained high after renal dialysis but returned to normal following renal transplantation.
The effect of malonyldialdehyde, a product of lipid peroxidation, on the deformability, dehydration and 51Cr‐survival of erythrocytes
Authors: Jain Sushil K. , Mohandas Narla , Clark Margaret R. , Shohet Stephen B.
Published: March 2008

Summary. Erythrocyte membrane lipid peroxidation has been reported to occur in various haemolytic anaemias. In the present study, treatment of human erythrocytes with malonyldialdehyde (MDA). a product of fatty acid peroxidation, induced membrane rigidity, cellular dehydration and reduced whole cell deformability. These effects of MDA were blocked by histamine and fluorescamine, which can act as alternate substrates for MDA. Additionally, reduced deformability of MDA‐treated rabbit cells was associated with shortened 51Cr survival in vivo. These findings suggest a biochemical basis for decreased survival of erythrocytes undergoing peroxidative damage of the membrane.
Monomeric IgG preparations for intravenous use inhibit platelet stimulation by polymeric IgG
Authors: Gross B. , Haessig A. , Luescher E. F. , Nydegger U. E.
Published: March 2008

Summary. Seven different immunoglobulin G (IgG) preparations for intravenous use were tested for their capacity to inhibit serotonin‐release from washed human platelets induced by IgG coupled with bis‐diazobenzidine. Using a reference standard consisting of the top third fraction of an ultracentrifuged gammaglobulin preparation for intramuscular use, two chemically treated preparations were less potent inhibitors than the standard while two preparations, one pH 4 treated and another albumin protected, were better inhibitors. A pepsin treated preparation was devoid of inhibitory capacity, whereas Fc fragments derived from human IgG were extremely efficient. Evidence is discussed that the inhibitory capacity is inversely correlated to the content of oligo‐ and/or dimeric IgG molecules.
Rapid proteolysis of unstable globins in human bone marrow
Authors: Vissers Margret C.M. , Winterbourn Christine C. , Carrell Robin W.
Published: March 2008

Summary. To determine whether human red cells contain a proteolytic system capable of rapidly degrading unstable proteins, the fate of pulse‐labelled puromycyl polypeptides was investigated. In erythroid bone marrow cells these unstable polypeptides were degraded to TCA‐soluble fragments with a mean half‐life of 4–5 min. However, in peripheral blood reticulotyes no proteolysis was observed, indicating that this activity declines as the red cell matures.
Reversal of marrow fibrosis in acute megakaryoblastic leukaemia after remission‐induction and consolidation chemotherapy followed by bone marrow transplantation
Authors: Mehta A. B. , Baughan A. S. J. , Catovsky D. , Goldman J. M. , Johnson S. A. , Galton D. A. G.
Published: March 2008

Summary. A young man with acute megakaryoblastic leukaemia was successfully treated with combination chemotherapy. A complete remission was obtained and further chemoradiotherapy followed by allogeneic marrow transplantation was administered. He is well and in complete remission of his disease 8 months after transplantation.
Nephrotoxicity in bone marrow transplant recipients treated with cyclpsporin A
Authors: Hows J. M. , Chipping P. M. , Fairhead S. , Smith J. , Baughan A. , Gordon‐Smith E. C.
Published: March 2008

Summary. Cyclosporin A (CyA) is a valuable post graft immunosuppressive agent in allogeneic bone marrow transplantation. The use of CyA is associated with a reduction in severity of graft versus host disease and improved marrow engraftment.

A major side effect of CyA is nephrotoxicity. In 33 patients studied during the first 4 weeks of therapy there is a close correlation between trough (12 h) serum cyclosporin A concentrations and plasma creatinine (r=0·93, P < 0·001) and urea (r=0·88, P < 0·001). Trough CyA serum concentrations of > 500 ng/ml are potentially nephrotoxic. Other risk factors for early nephrotoxicity in cyclosporin therapy are the concurrent use of aminoglycoside antibiotics (P=0·01) and hyperbilirubinaemia (P=0·01). Early nephrotoxicity can be prevented by maintaining trough CyA levels in the range 100–400 ng/ml.

During prolonged CyA therapy, cumulative renal impairment can occur and nephrotoxic episodes associated with microangiopathic peripheral blood changes and hypertension are seen in a minority of patients.
Fast migrating protein, immunochemically related to human factor VIII, studied by crossed immunoelectrophoresis in agarose
Authors: Thomas Kathy B. , Howard Margaret A. , Salem Hatem H. , Firkin Barry G.
Published: March 2008

Summary. A fast migrating protein (FMP) was detected by agarose radio‐crossed immunoelectrophoresis, in addition to factor VIII antigen (VII:RAg), using antiserum to human factor VIII (FVIII). FMP had partial immunochemical identity with FVIII, migrated as an α‐protein, and was distinct from α‐2 macroglobulin, fibronectin or IgM. FMP was precipitated by concanavalin A and was separable from the bulk of VII:RAg by ammonium sulphate fractionation. A significant amount of FMP was seen in normal serum (n= 12), plasma from patients with: (a) disseminated intravascular coagulation (n= 12) and (b) severe haemophilia A (n= 6). Trace amounts of FMP were observed in plasma from normal donors (n= 12), but neither VIII: RAg nor FMP was detectable in the plasma or serum from patients with severe von Willebrand's disease (n= 3). Freshly prepared cryoprecipitate contained trace amounts of FMP, similar to normal plasma, but increased levels were observed in antihaemophilic factor concentrates prepared for patient use. Significant levels of FMP were also seen in cryoprecipitate after storage at 4°C for 7 d and this generation of FMP was diminished by the addition of protease inhibitors. The presence of significant levels of FMP in situations where proteolytic enzymes may be activated and inhibition of its generation by protease inhibitors, suggest that this protein is produced by proteolytic action of enzyme(s) on the FVIII molecule.
Quantitative cytochemistry of blood neutrophils in acute myeloid leukaemia
Authors: Schofield K. P. , Stone P. C. W. , Stuart J.
Published: March 2008

Summary. Blood neutrophils were studied by quantitative cytochemistry in patients with acute myeloid leukaemia at diagnosis (17 patients), during remission (17 patients) and in relapse (seven patients). Scanning and integrating microdensitometry was used to quantify components of azurophilic granules (myeloperoxidase, chloroacetate esterase, β‐glucuronidase, acid phosphatase, acid mucosubstance) and also specific granules (lactoferrin). At diagnosis, neutrophil myeloperoxidase, chloroacetate esterase, and lactoferrin were significantly decreased, compared with normal neutrophils from 25 controls, with 13 of the 17 patients showing a partial or complete deficiency of at least one granule constituent. Five of seven patients, followed serially from remission into relapse, showed a fall in activity of azurophilic or specific granule components before overt blast cell infiltration of the marrow had occurred and this may predict relapse.