bluuebunny/crossref_metadata_2025 huggingface dataset:
Decreased Erythropoiesis: The Origin of the BCG Induced Anaemia in Mice
Authors: Marchal Gilles , Milon Geneviève
Published: March 2008

Summary.

Transient anaemia follows a high dose of viable BCG given intravenously to mice. The anaemia was dependent upon the dose, viability and the route of injection of the BCG. The evidence of an absolute anaemia was based on red cell mass decrease.

Serum iron concentration and red cell life span were unchanged in the first 14 d after BCG infection. The erythropoietin level increased 20‐fold. Despite the high level of this hormone and the appearance of increased erythropoiesis in the spleen, the production of erythrocytes was decreased presumably because of a decrease in numbers of erythropoietin‐responsive cells.
Acid α‐naphthyl Acetate Esterase Activity in Nucleated Red Cells of Human Fetal Liver
Authors: Lassila Olli
Published: March 2008

Summary.

Acid α‐naphthyl acetate esterase (ANAE) activity was investigated in nucleated red cells obtained from nine human fetal livers at 9‐24 weeks of gestation. A strong ANAE activity was demonstrated in fetal liver erythroblasts with a staining pattern of a perinuclear ring. Occasionally some polychromato‐philic normoblasts expressed a focal ANAE activity usually observed in human T‐lymphocytes. Some 50‐70% of erythroblasts in human fetal livers showed a ring‐like staining pattern. A slight decrease of ANAE positivity was observed in cells of fetal livers after 17 weeks of gestation.
A Clinical Trial on the Early Detection and Correction of Haemostatic Defects in Selected High‐risk Neonates
Authors: Turner T. , Prowse C. V. , Prescott R. J. , Cash J. D.
Published: March 2008

Summary. Low birth weight infants, preterm infants with respiratory distress syndrome and mature infants with asphyxia were screened for coagulation abnormalities as soon as possible after birth. Those with abnormal haemostasis were assigned to control or treatment groups and, in the latter case, were given prothrombin complex concentrate, cryoprecipitate or platelet concentrate as appropriate. This treatment was largely successful in correcting haemostatic defects but did not result in any change in mortality. There was no evidence of adverse or beneficial side‐effects of treatment.
EA Rosette Formation: a Simple Means to Increase Sensitivity of the Antiglobulin Test in Patients with Anti Red Cell Antibodies
Authors: Galili U. , Manny N. , Izak G.
Published: March 2008

Summary. Erythrocyte antibody (EA) rosette formation with the Fc‐receptor on the K‐562 erythro‐myeloid cell line was employed for the detection of subagglutinating amounts of Ig molecules bound to red cells. The sensitivity of this method exceeds that of the conventional direct and indirect antiglobulin tests without any alteration of the incubation media or pretreatment of red cells. The increased sensitivity did not diminish the specificity of the test, which can detect IgG, IgM and complement as well. This method may demonstrate the presence of antibodies on red cells in patients with suspected autoimmune haemolytic anaemia and negative antiglobulin test.
Microvesicles from Sickle Erythrocytes and their Relation to Irreversible Sickling
Authors: Allan David , Limbrick Anthony R. , Thomas Paul , Westerman Maxwell P.
Published: March 2008

Summary. Incubation of sickle (HbS) erythrocytes for periods up to 96 h leads to the formation of irreversibly sickled cells (ISCs) and to the release of spectrin‐free microvesicles similar to those derived from aged or Ca2+‐ionophore‐treated normal erythrocytes. The sickle microvesicles were somewhat larger than those from normal cells and showed minor differences in their membrane polypeptide composition. Sickle microvesicles were no different from their parent cells in their content of fetal haemoglobin. Neither microvesiculation nor formation of irreversibly sickled cells required the presence of Ca2+ in the medium but Ca2+ did accelerate both processes. Although in these prolonged incubations microvesiculation appeared to occur concomitantly with the formation of ISCs, it is not clear whether or not microvesiculation is a necessary prelude to irreversible sickling.
The Effect of Clinical Prostacyclin Infusions in Advanced Arterial Disease on Platelet Function and Plasma 6‐keto PGF1α Levels
Authors: Machin S. J. , Chamone D. A. F. , Defreyn G. , Vermylen J.
Published: March 2008

Summary. We have infused synthetic prostacyclin (PGI2) continuously for approximately 72 h at the maximum tolerated dose (ranging from 5 to 60 ng/kg/min) into nine patients with advanced arterial disease. Prior to the infusion seven out of nine patients had spontaneous platelet aggregation and five out of six patients tested had an abnormal circulating platelet aggregate ratio. During the infusion only one patient still had spontaneous aggregation and all the abnormal circulating platelet aggregate ratios returned to the normal range. However, none of the patients showed any suppression of ADP induced aggregation. The level of exogenous PGI2 required in vitro prior to the infusion to completely inhibit ADP induced aggregation was 5–10 ng/ml in three of the four patients tested. Ten healthy adults showed complete inhibition with 1 ng/ml of PGI2. It appears that the platelets of some patients with arterial disease are more resistant to the anti‐aggregating properties of PGI2.
Origin of Anti‐Thomsen‐Friedenreich (T) and Tn Agglutinins in Man and in White Leghorn Chicks
Authors: Springer Georg F. , Tegtmeyer Herta
Published: March 2008

Summary. Interest in anti‐Thomsen‐Friedenreich (T) antibodies has increased because of their significance in detection of and their possible interaction with human adenocarcinoma. The origin of anti‐T, which all humans possess, has not been ascertained. We determined here that anti‐T and ‐Tn agglutinins could readily be induced via the physiological intestinal route by an enteric bacterium, E. coli O86, which possesses T and Tn activities.

One dose of live E. coli O86 given in the drinking water to germfree chicks, who had no anti‐T and ‐Tn antibodies, resulted, in all birds, in formation of saline agglutinating anti‐T and ‐Tn antibodies as well as those detectable only by indirect agglutination. Antibody specificity was confirmed by adsorption on and elution from homologous human erythrocytes and for anti‐T also by haemagglutination inhibition. In contrast, control chicks raised under ordinary conditions did have anti‐T and ‐Tn prior to feeding E. coli O86.

In humans, six diarrhoeic and five healthy infants and the majority of 13 adults investigated were fed killed rather than live E. coli O86. All infants, but one, suffering from diarrhoea showed a significant increase (4‐fold) in anti‐T and/or anti‐Tn antibodies; in some, these antibodies were elicited de novo. All four adults with intestinal lesions had a significant increase of anti‐T and/or ‐Tn subsequent to ingestion of E. coli O86, as did five of nine healthy adults, but to a lesser extent. These findings support the immune nature of demonstrable levels of anti‐T and ‐Tn.
Thymidylate Synthesis in a Folate Deprived Cell Line
Authors: Marquet J. , Zittoun J. , Weynants C. , Zittoun R.
Published: March 2008

Summary. Abnormalities of the de novo and salvage pathways of thymidylate synthesis have been investigated in a folate deprived lymphoblastoid cell line. Impaired DNA synthesis was observed, with an increased percentage of cells in S and G2 phase, whereas the mitotic index was decreased.

Thymidylate synthesis along the salvage pathway was markedly increased, with a higher activity of thymidine kinase and higher uptake of 3H‐thymidine (3H‐TdR). The same abnormalities were observed when cells were treated with 5‐fluorodeoxyuridine or methotrexate.

The de novo pathway was slightly modified with a nearly normal incorporation of 3H‐6 deoxyuridine (3H‐UdR) and a moderate decrease of thymidylate synthetase activity; in contrast, the uptake of 3H‐dU was markedly inhibited in drug‐treated cells.

Preincubation with cold deoxyuridine (10−4 M) did not suppress the uptake of 3H‐TdR as efficiently as in control cells; with increasing concentration of dU to 10−2 M, this suppressive effect became almost complete. This high concentration of cold dU exerted a competitive inhibition on thymidine kinase.

The deoxythymidine triphosphate (dTTP) pool was increased in deficient cells and it was only slightly increased by addition of cold dU (10−4 M) in the culture medium whereas a substantial expansion of this pool was observed in control cells treated under the same conditions.

These data do not necessarily exclude a defect of thymidylate synthesis along the de novo pathway in the folate deficient cells. The normal incorporation of 3H‐dU could be explained by a decreased isotope dilution due to a reduced deoxyuridine monophosphate (dUMP) pool. This pool could be decreased by feedback inhibition by dTTP on some enzymatic activities, mainly deoxycytidylate deaminase. The enlarged dTTP pool which probably derives mainly from the salvage pathway could be poorly functional for DNA replication according to the model of compartmentation of DNA precursors.
A Proliferative Defect of Human Bone Marrow after Transplantation
Authors: Barrett A. J. , Adams J.
Published: March 2008

Summary. A technique which permits proliferation of CFU‐C or their precursors is described. Bone marrow cells from 10 normal donors showed an increase over a 2 week culture period of total nucleated cell counts, and of granulocyte colony and cluster forming cells. During culture an adherent layer of fibroblasts and monocyte‐macrophage cells formed. Bone marrow from eight patients taken between 2 weeks and 2 years after marrow transplantation did not show CFU‐C proliferation or increase in total cell counts in culture. This difference did not appear to be due to changes in the adherent cell layer formation, and suggests that grafted human bone marrow shows a proliferative defect similar to that described in mouse transplantation models.
Long‐term Abnormalities in T and B Lymphocyte Function in Children following Treatment for Acute Lymphoblastic Leukaemia
Authors: Layward Lorna , Levinsky Roland J. , Butler Margaret
Published: March 2008

Summary. T and B lymphocyte function as assessed by pokeweed mitogen stimulation and concanavalin A suppression was studied in 40 children with acute lymphoblastic leukaemia who were in remission and off all treatment. Both plasma cell and immunoglobulin production was reduced in off‐treatment patients, particularly in those who had received cyclophosphamide as part of their treatment. The ability to suppress immunoglobulin production was also significantly reduced in these patients. These abnormalities were long‐standing and were apparent up to 5 years after stopping treatment with no indication of a return to normality. No clinical correlations have been observed.
Ferritin Synthesis in Inflammation: II. MECHANISM OF INCREASED FERRITIN SYNTHESIS
Authors: Konijn A. M. , Carmel N. , Levy R. , Hershko C.
Published: March 2008

Summary. The mechanism of increased ferritin synthesis in inflammation was studied in rat livers 0—48 h after turpentine injection. A subcellular protein synthesizing system was employed in which the respective roles of cell sap factors and polysomes from normal and treated animals could be studied. Two waves of increased ferritin synthesis were found, an early wave with peak activity at 6 h of inflammation, and a second wave starting at about 24 h. The early wave of enhanced ferritin synthesis was associated with increased activity of cell sap factors. In contrast, the late enhancement of ferritin synthesis was characterized by increased polysomal activity as well as increased cell sap activity. These observations suggest a post‐transcriptional control mechanism for the early phase of enhanced ferritin synthesis in inflammation, and a transcriptional as well as post‐transcriptional control for the late phase of enhanced ferritin synthesis.
B Lymphocyte Function in B Cell Chronic Lymphocytic Leukaemia
Authors: Wolos Jeffrey A. , Davey Frederick R.
Published: March 2008

Summary. B‐enriched lymphocyte populations from patients with chronic lymphocytic leukaemia (CLL) were compared to B‐enriched lymphocyte populations from normal age‐matched controls for their ability to stimulate a proliferative response and to generate cytotoxic cells in allogeneic mixed lymphocyte cultures (MLC). The proliferative responses of MLCs were less than normal when the stimulating cells originated from B cells of patients with CLL. The degree of stimulation provided by the B cells from the patients with CLL inversely correlated with the peripheral white cell count. Lymphocytes from patients providing a poor stimulatory signal in the MLC tended to have less‘Ia‐like’antigen than lymphocytes giving a moderate level of stimulation. B lymphocytes from patients with CLL which stimulated poorly in the MLC also failed to generate specific cytotoxic cells even when provided with a normal proliferative trigger. These data suggest that B lymphocytes from cases of CLL with markedly elevated leucocyte counts may have a diminished concentration of both‘Ia‐like’and serum defined antigens.
Release of Iron by Resident and Stimulated Mouse Peritoneal Macrophages following Ingestion and Degradation of Transferrin—Antitransferrin Immune Complexes
Authors: Esparza Inmaculada , Brock J. H.
Published: March 2008

Summary. A method of loading macrophages from normal and inflammatory mouse peritoneal exudates with59Fe using59Fe,125I‐transferrin—antitransferrin immune complexes is described and the subsequent release of iron and degraded transferrin to the incubation medium has been studied. Release of iron occurred more rapidly from resident macrophages than from thioglycollate broth‐induced (stimulated) macrophages, but degradation of the125I‐transferrin in the immune complexes was faster in stimulated cells. A small percentage of the iron released was in the form of ferritin. Desferrioxamine (1 mM) increased the release of iron from both stimulated and resident macrophages, the effect being proportionally greater in the stimulated cells. Ascorbic acid (1 mM) had no effect on the release of iron, nor did the addition of apotransferrin (1 mg/ml) to the culture medium. These results support the concept of a blockade of iron release by reticuloendothelial cells in states of inflammation, and suggest that it may be a primary cause of the anaemia of chronic disease.
T‐cell function in chronic lymphocytic leukaemia
Authors: Whelan C. A. , Willoughby R. , McCann S. R.
Published: March 2008

Summary. Fifteen patients with chronic lymphocytic leukaemia (CLL) were studied. The diagnosis was made by a combination of clinical findings, peripheral blood morphology and cell surface markers. The Rai clinical staging (Rai et al, 1975) was used to classify all patients. Serum immunoglobulins, B and T cell lymphocytes, TG and TM cell populations were evaluated in all patients before commencing treatment.

No correlation was found between clinical staging and the presence or absence of hypogammaglobulinaemia. The ratios of TG to TM cells were abnormal in all patients and this abnormality paralleled the stage of the disease.

While there was a significant difference in the proliferative responses of highly purified T cells from patients and controls at 1 μg and 2 μg/ml of Concanavalin A (Con A) (P values =P < 0.005 and P < 0.01 respectively). There was no significant difference at dose 5 μg/ml.

The ability of T cells to suppress allogeneic PBMC responses to Con A was dependent on T cell purification procedures. The significance of the results and the possible role of T cells in the pathogenesis of CLL is discussed.
Modulation of leukaemia blast colony growth by steroid hormones
Authors: Taetle Raymond , Guittard Jean‐Pierre
Published: March 2008

The effect of steroid hormones on in vitro colony growth of human myelogenous leukaemia cells was examined. β‐Oestradiol, testosterone and 5‐β‐dihydrotesterone had little effect on HL60 promyelocytic leukaemia cells or blast colony forming cells (CFC) from eight patients with acute non‐lymphocytic leukaemia (ANLL). These compounds inhibited blast colony growth at very high concentrations (10−6‐10−4 M). In contrast, hydrocortisone (HC) had highly variable effects on blast colony growth. HL60 cells were resistant to HC at concentrations up to 10−4 M but blast CFC showed a wide range of response. Some patients were resistant to HC at all concentrations tested, while others were inhibited by concentrations as low as 10−8 M. The inhibitory effect of HC was also observed on 3H‐TdR incorporation by stimulated peripheral blood blasts. Inhibition by HC was not blocked by progesterone, suggesting this effect was not mediated by specific hormone receptors. These data indicate differences in the response of blast cells in ANLL and normal progenitors to steroid hormones. Cells from patients with ANLL display variable inhibition by HC which is probably not mediated by specific receptors.
Haemoglobin synthesis in CFU‐E‐derived erythroid colonies during sheep ontogeny
Authors: Darbre Philippa D.
Published: March 2008

The patterns of haemoglobin synthesis in erythroid colonies grown from sheep marrow CFU‐E of different ontogenic stages have been reported previously (Darbre et al, 1979) but only at a single fixed time point in culture. Culture of adult and late fetal progenitors enhanced γ chain synthesis whereas culture of earlier progenitors during the‘fetal switch’seemed to increase β‐chain synthesis. In order to try to explain these findings, patterns of growth and of haemoglobin synthesis in the colonies were examined over a time‐course of 6 d for all developmental stages. Colonies assessed later in vitro made proportionately less γ chain at every age. It was concluded that because fetal colonies mature more quickly in vitro than later and adult counterparts, and also because γ and β chain synthesis appeared to be asynchronous during sheep colony maturation, the results of assay at a single time point are not strictly comparable. The relevance of these findings to the study of the switch from fetal to adult haemoglobin synthesis is discussed.
GM‐CFC growth in chronic granulocytic leukaemia is not affected by a soluble inhibitor released by aplastic anaemia T‐cells or mitogen‐primed normal T‐lymphocytes
Authors: Frassoni Francesco , Podesta Marina , Van Lint Maria T. , Marmont ALlberto , Roosi Edoardo , Vimercati Renato , Bacigalupo Andrea
Published: March 2008

Peripheral blood (PB) and bone marrow (BM) cells were obtained from 12 patients with chronic granulocytic leukaemia (CGL) and cultured in agar for granulocyte macrophage colony formation (GM‐CFC). In addition PB and BM CGL cells were co‐cultured with T‐lymphocytes from patients with immune severe aplastic anaemia (SAA), or with T‐cells from healthy donors primed with pokeweed mitogen (PWM). The supernatants of SAA and PWM primed T‐cells were also added to CGL cells grown in agar. Normal marrow cells obtained from eight healthy donors were used to set up control cultures and co‐cultures. The results of this study indicate that, firstly, T‐cells derived from SAA patients and their supernatants suppress GM‐CFC growth of normal marrow cells but not of PB nor BM CGL cells, and, secondly, normal T‐cells primed with PWM and their supernatants suppress normal marrow GM‐CFC but not colony formation of BM nor PB CGL cells. These results provide further evidence that mediators which are effective in regulating normal myeloid progenitor cells fail to inhibit the in vitro growth of GM‐CFC from CGL patients.
Polyvinylpyrrolidone (PVP): a new precipitating agent for human and bovine factor VIII and fibrinogen
Authors: Casillas Gerardo , Simonetti Celia
Published: March 2008

The PVP precipitating properties for human and bovine factor VIII and fibrinogen were studied and a new technique for the fractionation of factor VIII and fibrinogen was developed. The precipitation was performed at different temperatures and different PVP concentrations and the best conditions for the technique were chosen. The technique consists of two steps: (a) precipitation of factor VIII from undiluted plasma with PVP and (b) washing of the precipitate with a glycine‐saline solution. The final concentrate contains 90% of the factor VIII and 20% of the fibrinogen of the original plasma.
‘Silent’β‐thalassaemia caused by a ‘silent’β‐chain mutant: the pathogenesis of a syndrome of thalassaemia intermedia
Authors: Fessas Phaedon , Loukopoulos Dimitris , Loutradi‐Anagnostou Aphrodite , Komis George
Published: March 2008

Summary.

In a Greek family three cases of β‐thalassaemia intermedia were diagnosed as resulting from the interaction of a typical high HbA2‐β‐thalassaemia with an atypical (silent) β‐thalassaemia gene. Following electrophoresis of globins on an acid‐urea‐Triton‐acrylamide system, an otherwise silent β‐like variant was revealed in the carriers of the atypical thalassaemia gene and in the intermediates; it amounted to 33% of the non‐α chains in the former and to c. 75% in the latter. The provisional name Hb Knossos is suggested for this abnormality.
Bone marrow reconstitution of lethally irradiated canines using autologous bone marrow fractions obtained by counterflow centrifugation‐elutriation
Authors: Jemionek J. F. , Monroy R. L. , MacVittie T. J. , Contreras T. J. , Espy S. B.
Published: March 2008

Summary.

Canine bone marrow fractionated by counterflow centrifugation‐elutriation results in three areas of nucleated cell recovery. Fraction 1 accounts for 50% of the total nucleated cells and 25‐40% of the total recovered CFU‐GM activity. Fraction 2 contains less than 2% of the total nucleated cells and less than 0‐2% of the CFU‐GM activity. Fraction 3 accounts for approximately 50% of the total nucleated cell recovery and 60‐75% of the total recovered CFU‐GM activity. Animal survival was not directly correlated with the levels of CFU‐GM activity in the transfused fractions. Autologous infusion of these fractions into irradiated canines (9 Gy, 0.1 Gy/min) resulted in distinct survival profiles. Canines receiving autologous fraction‐2 cells showed no haematological reconstitution, with death occurring on days 10‐11 post‐irradiation. Canines receiving autologous fraction‐3 cells showed limited myeloid repopulation of both the bone marrow and peripheral blood with a mean survival time for 24 d. Canines receiving autologous fraction‐1 cells showed complete haematological reconstitution after 48 d and long‐term survival. The data may indicate a separation or enrichment of pluripotential stem cells (fraction 1) from committed myeloid progenitor cells (fraction 3).
Cell lines derived from a human myelomonocytic leukaemia
Authors: Bradley T. R. , Pilkington G. , Garson M. , Hodgson G. S. , Kraft N.
Published: March 2008

Summary.

Two cell lines have been isolated from separate samples, taken at the same time, from the peripheral blood of a case of myelomonocytic leukaemia. One of these (CESS‐B) is a typical B cell line obtained after treatment of the peripheral blood white cells with Epstein‐Barr virus (EBV). The second line (RC‐2) is a monocyte‐macro‐phage type, originally isolated by 5 months continuous culture in the presence of colony stimulating factor (CSF). Subsequently, after further culture, this line has become autonomous (RC‐2A), no longer requiring CSF for maintenance of proliferation, whilst retaining the characteristics of the original line.
Sudan Black B positivity in acute lymphoblastic leukaemia
Authors: Tricot G. , Orshoven A. Broeckaert‐Van , Hoof A. Van , Verwilghen R. L.
Published: March 2008

Summary.

An adult patient with Sudan Black B (SB B) positive leukaemic lymphoblasts is described. Peroxidase and naphtol AS D chloroacetate esterase stains were negative. The diagnosis of acute lymphoblastic leukaemia (ALL) was based on morphology (FAB classification: L1), on immunological marker studies (cALL+, Tdt+, Ia+) and on electron microscopy, revealing blasts, compatible with lymphoblasts. Additional proof of the diagnosis of ALL were the diffuse lymphadenopathy and the rapid response to ALL chemotherapy.

Scattered azurophilic granules were present in some lymphoblasts; ultrastructurally multiple lysosomal inclusions were detected, containing small vesicles, sometimes in association with entrapped cytoplasmic organelles. The large amounts of phospholipids in these inclusions explain the Sudan Black B positivity. This type of ALL can easily be misdiagnosed as acute myeloid leukaemia.
Influence of high molecular weight factor VIII on the measurement of low molecular weight factor VIII procoagulant in different assay systems
Authors: Muntean Wolfgang , Hathaway Wm. E. , Montgomery Robert R.
Published: March 2008

Summary.

Factor VIII procoagulant activity (VIIIC) is exerted by a low molecular weight (LMW) moiety of the factor VIII molecule that can be separated from a high molecular weight (HMW) moiety by high ionic strength buffers. In this investigation the procoagulant activity of the LWM moiety of factor VIII prepared by immuno‐adsorbent chromatography and its relationship to the HMW moiety of haemophilic plasma was studied by means of different VIIIC assay systems and using different substrates in regard to their content of the HMW VIII moiety. LMW VIIIC was prepared by immunoadsorbent chromatography; HMW VIII without VIIIC was prepared by chromatographing cryoprecipitate from a coagulant antigen negative severe haemophiliac on 4% agarose. The LMW VIIIC obtained by immunoadsorbent chromatography gave higher VIIIC values when tested in the one‐stage partial thromboplastin time (PTT) system using von Willebrand's disease plasma as substrate than using haemophilic plasma as substrate. This finding was shown to be related to the HMW VIII measured as VIII related antigen (VIII: Ag) in the substrate plasmas. When the VIIIR: Ag was removed from the haemophilic substrate plasma by immunoadsorption, the VIIIC values obtained for the LMW VIIIC were higher. Also, adding HMW VIII purified from haemophilic plasma to the von Willebrand's disease substrate plasma resulted in lower VIIIC values for the LMW VIIIC in the PTT system.
Fibrinogen Chapel Hill II: Defective in reactions with thrombin, factor XIIIa and plasmin
Authors: Carrell Nadia , McDonagh Jan
Published: March 2008

Summary. Fibrinogen Chapel Hill II is a hereditary, abnormal fibrinogen which is characterized by poor substrate reactivity toward thrombin, factor XIIIa and plasmin. The patient has a low plasma level of clottable protein with normal antigen concentration, high amounts of fibrinogen related material in serum, and prolonged thrombin and reptilase clotting times. Fibrinopeptide release was decreased with both thrombin and ancrod, indicating that release of fibrinopeptide A from the abnormal fibrinogen was impaired. Sequence analysis indicated that the A peptide was normal. Light scattering indicated that the fibrils formed by thrombin were unusually short and thick. When clotted under crosslinking conditions γ dimers formed normally but α polymer formation was defective. Under conditions which yielded complete plasmin digestion of normal fibrinogen only half of the patient fibrinogen was degraded beyond the fragment X stage. The rate of fibrinopeptide release from patient fragment X and NH2‐terminal disulphide knot (N‐DSK) was similar to that from the fibrinogen, indicating that the defect was contained within the N‐DSK. A simple amino acid substitution could result in a conformational defect in the N‐DSK sufficient to perturb the reactions involving thrombin, factor XIIIa and plasmin and also polymerization.
Molecular aspects of the binding of absorbed iron to transferrin
Authors: Marx J. J. M. , Klein Gebbink J. A. G. , Nishiasto Takuji , Aisen P.
Published: March 2008

Summary. To study the molecular aspects of the binding of absorbed iron to plasma transferrin, 59Fe with high specific activity was administered via intragastric tube to iron‐deficient rabbits. The distribution of the absorbed 59Fe among the molecular forms of iron‐transferrin was analysed using urea‐polyacrylamide gel electrophoresis. Absorbed iron was bound to circulating transferrin one atom at a time. In four out of five animals, absorbed iron was predominantly bound to the site in the N‐terminal domain of the protein. Thus, the two sites of transferrin may differ in their ability to load absorbed iron.
A SINGLE‐STEP BACTERIAL BINDING ASSAY FOR THE CLASSIFICATION OF CELL TYPES WITH SURFACE ANTIGEN‐DIRECTED MONOCLONAL ANTIBODIES
Authors: Uchanaska‐Ziegler Barbara , Wernet Peter , Ziegler Andereas
Published: March 2008

Summary. A rapid and cheap single‐step bacterial‐binding assay for the detection of cell‐membrane molecules is described. It offers the opportunity to correlate antigen expression with cell morphology and cytochemistry and gives an indicagtion of the quantitiy of a cell surface molecule. The value of this procedure is demonstrated by the analysis of spontaneously grwoing and differentiating cultures of the human myeliod leukaemia line HL‐60 with monoclonal antibodies.
Haemostasis in heat stroke
Authors: Beard Michael E. J. , Hickton Christine M.
Published: March 2008

Summary. A patient with moderately severe heat stroke was studied with particular attention to changes in haemostasis. Activation of fibrinolysis, a probable failure of hepatic synthesis of coagulation factors and severe thrombocytopenia were observed, but there was no definite evidence for disseminated intravascular coagulation (DIC). It is suggested, however, that in heat stroke of greater severity the more extensive tissue breakdown may lead to DIC. If DIC is superimposed on the above changes in haemostasis it is likely to be both sudden and overwhelming.
Effect of hydrocortisone on the growth of human bone marrow fibroblasts
Authors: Minguell José J. , Martianez Jorge , Walter Tomas
Published: March 2008

Summary. Fibroblast‐like cells may be grown from human bone marrow. Adherent cultures show a logarithmic phase of growth lasting 6 d. Hydrocortisone (1 x 10‐8 M) decreases cell growth in 40‐60% when measured as cell number and DNA synthesis. Fibroblasts bind the hormone by a high affinity, saturable process showing a Kd of 2.2 x 10‐8 M and an average binding capacity of 92 fmol/mg protein. These findings suggest the involvement of glucocorticoids in the regulation and differentiation processes in bone marrow stromal cells.
Generation of CFU‐C suppressor T cells in vitro: V. A. MULTISTEP PROCESS
Authors: Bacigalupo Andrea , Podesta' Marina , Frassoni Francesco , Piaggio Giovanna , Van Lint Maria T. , Raffo Maria R. , And Mario Repetto , Marmont Alberto
Published: March 2008

Summary. Different cell fractions obtained from five patients with immune severe aplastic anaemia (SAA) in complete autologous haematologic reconstitution were tested for CFU‐c suppression. Bone marrow mononuclear cells (BMMC), but not peripheral blood mononuclear cells (PBMC), showed definite CFU‐c inhibitory activity. On the contrary, both peripheral blood and marrow E rosetting cells (E+) suppressed CFU‐c growth. The suppressor activity of PBE+ cells could not be rescued by adding back PBE‐ cells and/or PB adherent cells (AC). In addition, unfractionated PBMC exposed to sheep red blood cells (SRBC) suppressed CFU‐c growth. PBMC from normal donors exposed to SRBC had no suppressor activity.

This study suggests that CFU‐c suppressor T cells in the peripheral blood of SAA patients are in different activation state as compared to BM cells from the same patients, and also differ from normal PB cells. The identification of T cells with different requirements for in vitro activation in order to exhibit a suppressor activity, suggests that generation of suppressor cells is a multistep process, and this may have practical implications for in vitro assays designed to test for immune suppression of haematopoiesis.
Natural killer (NK) cell sensitivity of phorbol ester‐differentiated tumour cells correlates with disease activity in chronic B‐lymphocytic leukaemia
Authors: Tötterman Thomas H. , Gidlund Magnus , Nilsson Kenneth , Wigzell Hans
Published: March 2008

Summary. In a prospective study, 16 patients with chronic B‐lymphocytic leukaemia (CLL) were repeatedly investigated for clinical and laboratory signs of disease activity during a period of 18 months. The leukaemic cells were induced to differentiate in vitro using a phorbol ester, TPA. Two differentiation‐associated properties of the tumour cells, i.e. the amount of cytoplasmic immunoglobulin (C‐IgM) and the increased sensitivity to lysis by natural killer (NK) cells, were expressed in parallel and both parameters were positively correlated with the clinical activity of leukaemia. Phorbol ester‐induced phenotypic changes of the leukaemic cells may therefore be used for the in vitro monitoring of disease activity in CLL. The present markers may, in addition, be useful in molecular analysis of B cell differentiation.
Globin gene mapping in normal Hb A2 types of β‐thalassaemia
Authors: Kanavakis E. , Metaxotou‐Mavromati A. , Kattamis C. , Aksoy M. , Weatherall D. J. , Wood W. G.
Published: March 2008

Globin‐gene mapping of DNA from 13 families with normal Hb A2β‐thalassaemia (both type 1 and type 2) failed to detect any difference from normal in their globin‐gene arrangement. We conclude that deletions such as those responsible for γβ‐thalassaemia or a ‘silent’ Hb Lepore are not responsible for this type of β‐thalassaemia in Greece.
Small doses of ARA‐C in the treatment of acute myeloid leukaemia: differentiation of myeloid leukaemia cells?
Authors: Housset M. , Daniel M. T. , Degos L.
Published: March 2008

Three patients with acute myeloid leukaemia were treated with small doses of ARA‐C (10 mg/m2/12 h, subcutaneous injections) and complete remission was obtained. The small doses of ARA‐C, the progressive evolution, the absence of aplasia before remission, the simultaneous presence of normal islets of promyelocytes and leukaemic myeloblasts, favour a differentiating role for the drug rather than an antimitotic effect.
Megakaryocyte cultures in the chronic phase and in the blast crisis of chronic myeloid leukaemia: studies on the differentiation of the megakaryocyte progenitors and on the maturation of megakaryocytes in vitro
Authors: Vainchenker W. , Guichard J. , Deschamps J. F. , Bouguet J. , Titeux M. , Chapman J. , McMichael A. J. , Breton‐Gorius J.
Published: March 2008

Megakaryocyte (MK) colony formation has been studied in the chronic phase and in the blast crisis of chronic myeloid leukaemia (CML). Blood cells were grown in plasma clot for 13 d. MKs were subsequently identified by immunofluorescent techniques using two monoclonal antiplatelet antibodies (AN51 and J15). The maturation process was studied by ultrastructural methods. A marked increase in the number of circulating CFU‐MK was observed in all the 10 cases studied prior to chemotherapy (70‐fold increase per ml of blood). No significant modification in the regulation of MK colony formation as compared to that of normal subjects was observed. The predominant abnormality in maturation in culture was the occurrence of many hypoploid MKs (microMKs). However, the cytoplasmic maturation of the MKs was identical to that of normal subjects with occasional platelet shedding. Since microMKs predominated in some patients, scoring of MK colonies in CML necessitated immunofluorescent labelling to permit identification of MKs. During the blast crisis, MK colony formation occurred in four out of five patients with an extremely high plating efficiency in the case of promegakaryoblastic transformation. In contrast, MK colonies could not be grown from blood samples of patients with acute leukaemia, including two cases of promegakaryoblastic leukaemia. Maturation of MKs in blast crisis was identical to that of the chronic phase. Furthermore, after short periods of culture in liquid medium, circulating promegakaryoblasts from spontaneously to become large MKs exhibiting demarcation membranes and α‐granules, while those from two cases of megakaryoblastic leukaemias did not mature. In consequence, the blast crisis of CML exhibits a different culture pattern from acute leukaemia. These results suggest, therefore, that the acute transformation of CML cannot be simply explained by clonal changes and that environmental and regulatory factors could be also involved.
Antithymocyte globulin treatment of severe aplastic anaemia
Authors: Miller Wesley J. , Branda Richard F. , Flynn Patrick J. , Howe Robert B. , Ramsay Norma K. C. , Condie Richard M. , Jacob Harry S.
Published: March 2008

Summary.

Nineteen patients with severe aplastic anaemia were treated with antithymocyte globulin. Ten patients obtained remissions (transfusion independent, at least 45000 platelets and 2000 PMN/mm3) within 2–3 months and continue in remission 5–35 months after antithymocyte globulin. Ages of responders ranged from 17 to 71. Complications of antithymocyte globulin included arthralgias, rash, serum sickness, angioedema and fever. Two patients died during, two shortly after, and one 10 months after therapy. One patient with a previous remission following antithymocyte globulin relapsed and achieved a second remission with retreatment. Previous androgen therapy did not affect outcome since two of four patients with and eight of 15 patients without previous androgen therapy achieved remission with ATG. Treatment with antithymocyte globulin is a promising alternative to bone marrow transplantation in the treatment of severe aplastic anaemia.
The effect of stasis on the venous endothelium: an ultrastructural study
Authors: Thomas D. P. , Merton R. E. , Hockley D. J.
Published: March 2008

Summary.

In carefully dissected neck veins, no evidence was found of platelet adherence to the vessel wall or leucocyte migration. However, 30–60 min of total stasis led to polymorphonuclear leucocytes sticking to the endothelium and their subsequent migration. This migration across the vessel wall resulted from stasis and not the trauma of dissection. Adherence and migration of leucocytes did not cause gross endothelial cell damage or desquamation within the observed period of stasis and there was no associated platelet adherence following restoration of blood flow. Thus leucocyte migration does not impair the non‐thrombogenicity of the endothelium in acute experiments.
Blood neutrophil function in primary myelodysplastic syndromes
Authors: Boogaerts M. A. , Nelissen V. , Roelant C. , Goossens W.
Published: March 2008

Summary.

Ten different tests of blood neutrophil function were studied in 20 patients with primary myelodysplastic syndromes (PMDS). The patients were selected according to the new diagnostic criteria for PMDS of the FAB‐cooperation group. Impairments of granulocyte functions were found in all patients. Moreover, several steps in the mobilization of granulocytes at the site of injury seemed to be affected: decreased adhesion (P < 0–05), deficient chemotaxis (P < 0.05), decreased enzyme content (P < 0.001), ‘slower’chemiluminescence (P < 0005), decreased phagocytosis (P < 0.05) and impaired microbicidal capacity (P < 0.025). No significant correlation between disease category and severity of granulocyte dysfunction was discerned, though an increasing number of blasts was associated with more severe granulocytic disability. Results in seven patients with abnormal karotypes were not significantly different from 13 others with normal karyotypes. Our results indicate that defects in blood neutrophil function are a common feature in PMDS and might account for the increased frequency of infection in these patients.
T‐lymphocytes escape membrane defect in paroxysmal nocturnal haemoglobinuria
Authors: Cooper Michael R. , Currie Mark S. , Rustagi Pradip K. , Logue Gerald L.
Published: March 2008

Summary.

Erythrocytes, granulocytes and platelets from patients with paroxysmal nocturnal haemoglobinuria (PNH) are abnormally sensitive to lysis by complement. We studied T‐lymphocytes from PNH patients for abnormal complement lysis sensitivity. T‐lymphocytes free of other contaminating blood cells were prepared by sedimentation, nylon wool filtration, and density gradient centrifugation. The lymphocytes were then labelled with 51Cr and lysis induced by antithymocyte globulin and rabbit complement. PNH lymphocytes were no more susceptible to complement‐mediated lysis than lymphocytes from normal individuals. The unusual sensitivity of PNH erythrocytes could still be demonstrated when rabbit serum was a source of complement so the lack of any difference in the sensitivity of normal and PNH lymphocytes was probably not attributable to the inability of rabbit serum to elicit the membrane defect. PNH erythrocytes and granulocytes also acquire more membrane‐bound C3 when human complement is activated. Therefore we also searched for increased membrane C3 binding on PNH lymphocytes using anti‐I antibody and human serum as a complement source. C3 binding was measured using 125I labelled monoclonal mouse anti‐human C3. While we verified increased membrane C3 binding on PNH granulocytes during complement activation we were unable to show similar differences between PNH and normal T‐lymphocytes. Thus PNH T‐lymphocytes do not share the membrane abnormalities of PNH erythrocytes and granulocytes.
Normalized natural killer (NK) cell activity in long‐term remission of acute leukaemia
Authors: Yoda Yasuhiro , Abe Tsukasa , Tashiro Akio , Hirosawa Shinsaku , Kawada Kenichi , Onozawa Yasusuke , Adachi Yamao , Shishido Hideo , Nomura Takeo
Published: March 2008

Summary.

NK cell activity was studied in 28 cases of acute leukaemia in remission. It was found that patients in long‐term remission for 5 years or more had normal NK cell activity but those in remission for less than 5 years had significantly lowered activity. This suggests that patients with acute leukaemia had not been constitutionally deficient in NK cell activity. Low activity was augmented but not fully restored by interferon pretreatment in vitro. Therefore, the low NK cell activity observed during remission is not solely attributable to inadequate secretion of interferon in the host. It is suggested that the production of NK cells in bone marrow is impaired, or the function of NK cells is suppressed, by the leukaemia itself or by chemotherapeutic agents. Increased consumption of NK cells is also a possibility.
Bone marrow fibroblast function in relation to granulopoiesis in aplastic anaemia
Authors: Gordon M. Y. , Gordon‐Smith E. C.
Published: March 2008

Summary. Fibroblasts grown from the bone marrow of normal individuals and incubated with colony‐stimulating factor (CSF) enhance the stimulating activity of this CSF on granulocyte‐macrophage colony‐forming cell (GM‐CFC) proliferation.

For this study, the ability of fibroblast monolayers grown from the marrows of six patients with severe aplastic anaemia to increase the activity of CSF has been compared with the activity of normal fibroblasts. Tests of this function showed subnormal CSF‐enhancing activity by fibroblasts grown from the marrows of three of the six aplastic patients.

Since cultured bone marrow fibroblasts are thought to represent an important component of the haemopoietic microenvironment, the results suggest that some of the patients had a microenvironmental abnormality at the time of study.

One of the patients whose fibroblasts were abnormal was reinvestigated after he had been given a graft of allogeneic bone marrow cells. His post‐graft fibroblast function was normal, showing that the abnormality was reversible.
Effect of structural analogues of PAF‐acether on platelet desensitization
Authors: Keraly C. Lalau , Coëffier E. , Tencé M. , Borrel M. C. , Benveniste J.
Published: March 2008

Summary. The 1‐O‐alkyl‐2‐O‐acetyl‐sn‐glyceryl‐3‐phosphorylcholine (PAF‐acether) aggregates rabbit platelets and desensitizes them to a second challenge with the same agonist but not to arachidonic acid. The desensitizing activities of 14 analogues of PAF‐acether were explored with particular attention to the dose‐response dependency of the desensitization process. PAF‐acether was 500‐fold more active than its 1‐O‐acyl analogues. The 2‐lyso PAF‐acether was inactive and the PAF‐acether enantiomer 2000 times less effective than the natural isomer, thus confirming the importance of the presence and steric position of the 2‐acetate group. The desensitizing activities of the 2‐propionyl and the 2‐butyryl analogues were close to that of PAF‐acether. Substituting an ether to an ester bond at the 2‐position indicated that the number of carbon of the 2‐substituant seems more determinant than the nature of the linkage for the desensitizing process. Indeed, the 2‐ethoxy and the 2‐methoxy analogues were 87 and 5000 times less active than PAF‐acether respectively. The presence of methyl groups on the nitrogen base is also critical to desensitize platelets. The desensitizing potency of the tested phospholipids was always identical to their aggregating efficiency. It is suggested that these compounds activate cells through a common mechanism.
Cyclosporin A associated nephrotoxicity in the first 100 days after allogeneic bone marrow transplantation: three distinct syndromes
Authors: Atkinson K. , Biggs J. C. , Hayes J. , Ralston M. , Dodds A. J. , Concannon A. J. , Naidoo D.
Published: March 2008

Summary. Thirty‐six patients received allogeneic (34) or syngeneic (two) bone marrow transplants as treatment for severe aplastic anaemia or acute leukaemia. Nineteen of the allogeneic recipients received methotrexate (MTX) and 15 received cyclosporin A (CyA) as the predominant immunosuppressive agent to minimize graft‐versus‐host disease (GVHD) post transplant. In the first 100 d post transplant renal dysfunction was much less frequent in the MTX recipients than in the CyA recipients who exhibited three distinct syndromes of nephrotoxicity: most commonly, CyA recipients developed asymptomatic azotaemia, proteinuria, urinary casts, impaired urinary concentrating ability and hypertension. Secondly, two CyA recipients developed acute reversible renal failure precipitated by systemic bacterial infection which required dialysis and in which the kidney was the sole target organ; thirdly, two recipients of HLA‐genotypically non‐identical grafts developed a rapidly progressive fatal syndrome with multiple organ involvement including lung, brain and kidney which clinically and histologically resembled thrombotic thrombocytopenic purpura.
Anaemia in myelofibrosis: its value in prognosis
Authors: Njoku O. S. , Lewis S. M. , Catovsky D. , Gordon‐Smith E. C.
Published: March 2008

Summary. Forty‐four patients with myelofibrosis were investigated in our hospital in the period 1971–81. Their clinical, laboratory and radioisotope parameters were analysed. The direct correlation between plasma volume and splenic red cell pool has highlighted the role of the spleen in the dilutional anaemia seen in myelofibrosis. 52Fe quantitation enabled us to show that the bone marrow contributes relatively more to effective erythropoiesis than the extramedullary sites. The prognostic value of changes in plasma volume and bone marrow 52Fe activity has been demonstrated. We have shown that the Hb: reticulocyte relationship at diagnosis can be used to recognize probable stages of the disease and provides a useful prognostic determinant.