bluuebunny/crossref_metadata_2025 huggingface dataset:
RNAi‐mediated resistance to diverse isolates belonging to two virus species involved in Cassava brown streak disease
Authors: PATIL BASAVAPRABHU L. , OGWOK EMMANUEL , WAGABA HENRY , MOHAMMED IBRAHIM U. , YADAV JITENDER S. , BAGEWADI BASAVARAJ , TAYLOR NIGEL J. , KREUZE JAN F. , MARUTHI M. N. , ALICAI TITUS , FAUQUET CLAUDE M.
Published: July 2010

Cassava brown streak disease (CBSD) is emerging as one of the most important viral diseases of cassava (Manihot esculenta) and is considered today as the biggest threat to cassava cultivation in East Africa. The disease is caused by isolates of at least two phylogenetically distinct species of single‐stranded RNA viruses belonging to the family Potyviridae, genus Ipomovirus. The two species are present predominantly in the coastal lowland [Cassava brown streak virus (CBSV); Tanzania and Mozambique] and highland [Cassava brown streak Uganda virus (CBSUV); Lake Victoria Basin, Uganda, Kenya and Malawi] in East Africa. In this study, we demonstrate that CBSD can be efficiently controlled using RNA interference (RNAi). Three RNAi constructs targeting the highland species were generated, consisting of the full‐length (FL; 894 nucleotides), 397‐nucleotide N‐terminal and 491‐nucleotide C‐terminal portions of the coat protein (CP) gene of a Ugandan isolate of CBSUV (CBSUV‐[UG:Nam:04]), and expressed constitutively in Nicotiana benthamiana. After challenge with CBSUV‐[UG:Nam:04], plants homozygous for FL‐CP showed the highest resistance, followed by the N‐terminal and C‐terminal lines with similar resistance. In the case of FL, approximately 85% of the transgenic plant lines produced were completely resistant. Some transgenic lines were also challenged with six distinct isolates representing both species: CBSV and CBSUV. In addition to nearly complete resistance to the homologous virus, two FL plant lines showed 100% resistance and two C‐terminal lines expressed 50–100% resistance, whereas the N‐terminal lines succumbed to the nonhomologous CBSV isolates. Northern blotting revealed a positive correlation between the level of transgene‐specific small interfering RNAs detected in transgenic plants and the level of virus resistance. This is the first demonstration of RNAi‐mediated resistance to CBSD and protection across very distant isolates (more than 25% in nucleotide sequence) belonging to two different species: Cassava brown streak virus and Cassava brown streak Uganda virus.
AN ADDENDUM ON MATHEMATICAL TREATMENT OF THE DATA
Authors: Hartmann N. R. , Scheufens E. E.
Published: July 2010

Summary.

A case of acute lymphoblastic leukaemia has been studied by in vivo labelling with [3H]thymidine at two stages in the disease, first at the time of diagnosis before treatment and again at the first relapse. The study procedure was identical on both occasions.

At relapse, 4 days after discontinuation of 6‐mercaptopurine maintenance treatment, remarkable differences were observed as compared to the findings at diagnosis with respect to the curve of labelled mitoses, the mitotic index, and the frequency of cells in G2. These results suggest a larger spread of individual cell cycle phase times in relapse than at diagnosis. It cannot be decided whether this depends on the preceding chemotherapy or the evolution of the disease. Whatever the mechanism, the increased cytokinetic heterogeneity in relapse represents an obstacle when cell cycle phase dependent chemotherapy is attempted.

In addition, the transition of cells between pools of blast cells of different cell size was studied by measuring the size of labelled and unlabelled cells as a function of time after flash‐labelling. These data show that the small blast cells become labelled concomitantly with the division of the large, primarily labelled cells, that at least some small blast cells can replenish the group of large cells, and that a fraction of very small blast cells do not divide for a very long period of time.
Quantitative Aspects of the Destruction of Red Cells Sensitized with IgG1 Autoantibodies: An Application of Flow Cytofluorometry
Authors: Van Der Meulen F. W. , De Bruin H. G. , Goosen P. C. M. , Bruynes E. C. E. , Joustra‐Maas C.J. , Telkamp H. G. , Kr. von dem Borne A. E. G. , Engelfriet C. P.
Published: July 2010

Summary. The purpose of this study was to determine whether quantitative or qualitative factors are of major importance in the destruction of red cells sensitized with incomplete warm autoantibodies of subclass IgG1.

To that end, the relative amount of IgG1 antibody present on the red cells of patients with autoantibodies of this subclass only, was measured by means of continuous flow cytofluorometry. This method appeared to give an idea of the amount of antibody on red cells and was reproducible. The intensity of the fluorescence of patient's red cells, measured after incubation with a FITC‐labelled anti‐IgG1, was compared with the presence or absence of signs of increased haemolysis in vivo and the cytotoxic activity of normal monocytes towards these red cells in vitro.

It appeared that it was predominantly the amount of IgGl autoantibody that determined whether or not these antibodies induced haemolysis in vivo or cytotoxicity of monocytec in vitro. This was also true with methyldopa‐induced IgG1 autoantibodies.
Relationship between Fibrinogen Binding and the Platelet Glycoprotein Deficiencies in Glanzmann's Thrombasthenia Type I and Type II
Authors: Lee Helen , Nurden Alan T. , Thomaidis Annick , Caen Jacques P.
Published: July 2010

Summary. We have studied the fibrinogen binding to the platelets of four type I thrombasthenic, two type II thrombasthenic patients and 15 normal subjects. The amount of [125I]fibrinogen bound to washed platelets in the presence or absence of 10 μM ADP was measured and the results expressed as the percentage of the total radioactivity added. The mean value for specific binding to the platelets of normal subjects was 3–7 (range 1–7·25) at 15 min after the addition of ADP. Type I thrombasthenic patients lack glycoproteins (GP) IIb and IIIa in the platelet membrane and intra‐platelet fibrinogen. In all four patients studied, little or no specific fibrinogen binding was detected. Platelets of type II thrombasthenic patients contain detectable, although markedly reduced amounts of GP IIb and IIIa, and subnormal or normal levels of fibrinogen. In these platelets, significant specific fibrinogen binding was observed, the values being slightly below or at the lower end of the normal range. Fibrinogen binding to normal platelets was also measured after the addition of an IgG purified from the serum of a multi‐transfused thrombasthenic type I patient. This antibody was previously shown to be directed against the GP IIb/IIIa complex as located by crossed immunoelectrophoresis. Specific fibrinogen binding was blocked and the extent of the inhibition was directly proportional to the concentration of the antibody added.
Cytogenetics and Granulopoietic Effects of Bone Marrow Fibroblastic Cells in Fanconi's Anaemia
Authors: Greenberg Bernard R. , Wilson Floyd D. , Woo Linda , Knox Susan , Jenks Helen , Taplett Janet
Published: July 2010

Summary. To determine if abnormalities exist in the bone marrow stroma in Fanconi's anaemia, we studied the cytogenetic composition of in vitro bone marrow fibroblastic cells (FC) from a patient with this disorder and compared it to those obtained from skin fibroblasts, peripheral blood lymphocytes, and direct bone marrow preparations. The presence of granulocytic progenitors in bone marrow and T lymphocyte colonies in peripheral blood was also determined in addition to the ability of this patient's FC to stimulate normal granulocytic progenitors. We found that the FC had far fewer chromosomal abnormalities and stimulated normally the growth of granulocyte colonies. Granulocyte progenitors were not found, but T lymphocyte colonies were abundant. These results support the concept that a defect in haematopoietic stromal elements is not responsible for the aplasia developing in the disorder.
Effect of Low‐dose Irradiation on Pregnant Mouse Haemopoiesis
Authors: Weinberg Sheila R. , McCarthy Emmeline G. , MacVittie Thomas J. , Baum Siegmund J.
Published: July 2010

Summary. The effects of low‐dose gamma radiation to haemopoietic progenitor cell compartments of the marrow and spleen of virgin female mice and pregnant mice were studied. Microplasma clot cultures were used to assess burst‐forming uniterythroid (BFU‐E) and colony‐forming unit‐erythroid (CFU‐E) activity, and double‐layer agar cultures were established to evaluate granulocyte‐macrophage colony‐forming cell (GM‐CFC) and macrophage colony‐forming cell (M‐CFC). The apparent shift in maternal erythropoiesis from the bone marrow to the enlarged spleen was reflected by an increase in the numbers of CFU‐E and BFU‐E per spleen and a concomitant decrease in CFU‐E and BFU‐E per femur. Whereas maternal GM‐CFC values per femur increased 36%, maternal GM‐CFC per spleen increased by 172% compared to virgin values. There was a greater decrease in M‐CFC per spleen than per femur in the pregnant animal when values were compared to the virgin animal. Total‐body irradiation to the day‐10·5 pregnant mouse caused a further suppression of day‐14·5 medullary erythropoiesis (i.e. decreased CFU‐E values) compared to the response of the virgin female mouse. An ability of the maternal spleen to support further compensatory erythropoiesis following increasing doses of radiation was demonstrated. 4 d after 1·0 Gy exposure, maternal values for GM‐CFC per femur or spleen decreased to nonirradiated virgin mice values. M‐CFC per maternal femur decreased following 1·5 Gy, but M‐CFC per spleen appeared to be unaffected with doses from 0·5 to 2·0 Gy.
Abnormalities of T‐Cell Subsets in Patients with Neutropenia and an Excess of Lymphocytes in the Bone Marrow
Authors: Linch D. C. , Cawley J. C. , Worman C.P. , Galvin M. C. , Roberts B. E. , Callard R. E. , Beverley P. C. L.
Published: July 2010

Summary. Morphological, surface‐marker and culture studies were performed in three patients who had in common neutropenia and an excess of lymphocytes in the bone marrow, but who differed from each other in certain regards such as age and peripheral lymphocyte numbers. All three patients had an increased proportion of E‐rosetting cells bearing Fc receptors for IgG. These cells were shown to be T lymphocytes by staining with monoclonal anti‐T‐cell sera and their subset phenotype was established as OKT4− OKT8+; in addition, a variable proportion of the cells was Ia+. The functional significance of this expanded subpopulation of T cells was not clear, but it was shown that they did not inhibit colony formation in vitro.
Two Platelet Aggregation Inhibitors in Tsetse (Glossina) Saliva with Studies of Roles of Thrombin and Citrate inin VitroPlatelet Aggregation
Authors: Mant Michael J. , Parker Kenneth R.
Published: July 2010

Summary.

Two inhibitors of platelet aggregation have been identified in saline extracts ofGlossina morsitans(tsetse) salivary glands. A protein fraction (MW > 30000) inhibited primary and secondary aggregation to ADP, secondary aggregation to adrenalin, and aggregation to collagen. It also caused disaggregation of platelets stimulated by ADP and adrenalin. These properties could be explained by ADP hydrolytic activity. A previously identified antithrombin fraction (MW 11 000‐13 000) abolished thrombin‐induced aggregation. It did not affect platelet aggregation to ADP, adrenalin or collagen, as compared to aggregation in citrate, when used as sole anticoagulant for platelet rich plasma or when added to citrated platelet rich plasma. These results fail to support hypotheses (i) that thrombin plays an important role in platelet aggregation by other agonists and (ii) that secondary platelet aggregation is an artefact induced by citrate. It is proposed that these inhibitors may be important in maintaining mouthpart and crop patency during feeding. Their discovery suggests that other arthropods may also have antiplatelet agents in their saliva which might be entomologically important, provide useful tools for platelet studies, and be of potential therapeutic interest.
Carrier Detection in Haemophilia A by Immunological Measurement of Factor VIII Related Antigen (VIIIRAg) and Factor VIII Clotting Antigen (VIIICAg)
Authors: Peake I. R. , Newcombe R. G. , Davies B. L. , Furlong R. A. , Ludlam C. A. , Bloom A. L.
Published: July 2010

Summary.

23 obligate carriers of mild and severe haemophilia A and 26 normal females were bled on three occasions, and their plasmas assayed for procoagulant factor VIII (VIIIC), factor VIII related antigen (VIIIRAg) and factor VIII clotting antigen (VIIICAg). A comparison of the ratios VIIIC/VIIIRAg and VIIICAg/VIIIRAg indicated that, although the two ratios gave the same proportional misclassification of carriers as normals (four of 23), the latter ratio showed greater discriminatory power when an unequal variances predictive method was used to calculate likelihood ratios (for carrier status). This greater power was shown to be due to a greater reproducibility between visits for the VIIICAg/VIIIRAg ratio. Discrimination was considerably better when the median of the three median values for each variable was analysed, compared to the median value obtained at the first visit. There was also no statistical difference between VIIICAg/VIIIRAg (or VIIIC/VIIIRAg) ratios obtained from carriers of severe compared to mild haemophilia.
EFFECT OF HYDROCORTISONE ON LONG‐TERM HUMAN MARROW CULTURES
Authors: Suda T. , Dexter T. M.
Published: July 2010

Summary.

In human long‐term marrow cultures a relatively high concentration of hydrocortisone (10−6 M or more) is necessary for the development and subsequent maintenance of some cellular components in the adherent cell layer. However, such concentrations of hydrocortisone seem to be inhibitory for the production of granulocyte/macrophage progenitor cells (GM‐CFC). In further attempts to improve these cultures, therefore, it is important to maintain a balance between a concentration of hydrocortisone which is necessary for the development of a competent adherent layer and the concentration which is inhibitory to the production of GM‐CFC.
Bone marrow histology in myeloma: its importance in diagnosis, prognosis, classification and staging
Authors: Bartl R. , Frisch B. , Burkhardt R. , Moghadam A. Fateh‐ , Mahl G. , Gierster P. , Sund M. , Kettner G.
Published: July 2010

Summary.

A study has been made of 420 bone marrow biopsies from patients with multiple myeloma (220), idiopathic monoclonal gammapathy (50), reactive plasmacytosis (42) and solitary plasmacytoma (22). Histology and immunohistological parameters were more reliable than cytology in distinguishing a reactive from a neoplastic plasmacytosis. Histological variables were correlated with the clinical features of the patients to determine the factors which were of value in predicting prognosis. Plasma cell maturity and the extent of infiltration in the biopsy by myeloma cells proved to be highly significant in predicting the duration of survival. On the basis of these criteria multiple myeloma was classified into two types: plasmacytic of low‐grade malignancy and plasmablastic of high‐grade malignancy; and into three stages which accurately reflected the progression of the disease. We conclude that a bone biopsy provides useful information for the diagnosis, classification and staging of patients with multiple myeloma.
The differential ultrastructural localization of immunoglobulin heavy and light chains in human haematopoietic cell lines
Authors: Newell D. G. , Hannam‐Harris A. , Karpas A. , Smith J. L.
Published: July 2010

The ultrastructural localization of immunoglobulin heavy and light chains has been investigated in nine haematopoietic cell lines, using a technique which involves the treatment of lightly prefixed cells with saponin to allow penetration of the antibody‐peroxidase conjugate. The synthesis and secretion of immunoglobulin was also studied in these cell lines.

Immunoglobulin was found to be localized in the cisternae and on the membranes of the rough endoplasmic reticulum, perinuclear space and/or Golgi apparatus. In each case staining for heavy chains was weak or absent in the perinuclear space while staining for light chains was usually strong. Additionally in three cell lines immunoperoxidase staining indicated that heavy chains were absent from the Golgi apparatus despite the observed presence of light chains in the Golgi apparatus and the secretion of combined immunoglobulin into the supernatant. The results obtained suggest compartmentalization of the synthesis of light and heavy chains and indicate that the technique of immunoelectron microscopy may significantly contribute to an understanding of the mechanisms involved in immunoglobulin synthesis, intracellular transport and secretion.
Treatment of congenital antithrombin III deficiency with concentrates
Authors: Mannucci P. M. , Boyer C. , Wolf M. , Tripodi A. , Larrieu M. J.
Published: July 2010

Antithrombin III concentrates were administered to a patient with hereditary AT III deficiency undergoing orthopaedic surgery. The plasma AT III level (heparin cofactor activity) was maintained at values in excess of 100% postoperatively. A mean dose of 0.74 u/kg (range 0.64—0.83) of AT III resulted in a 1% increase in AT III level after concentrate infusion. The half‐life of AT III (mean 66 h; range 58‐76) was shorter during surgery and in the early postoperative period suggesting an increased consumption of the inhibitor. At III antigen was consistently higher than heparin cofactor activity during the first 24 h after concentrate infusions. This discrepancy was associated with the presence in the patient's plasma of an AT III fraction showing a slower mobility than the main AT III peak on two‐dimensional immunoelectrophoresis in the presence of heparin. A biological assay is preferred to an immunological assay in monitoring replacement therapy with AT III concentrates.
Rh immunization facilitated by passively‐administered anti‐Rh?
Authors: Contreras Marcela , Mollison P. L.
Published: July 2010

Summary. The response to 0·8 ml of D‐positive (ccDEE) red cells injected i.v., together with 1 μg anti‐D i.m., was studied in 13 D‐negative male volunteers (test subjects); the red cells were labelled with 51Cr and were found to have a mean survival time of 24 d, after correction for Cr elution. Within the following 5 months, nine of the 13 subjects made anti‐D, compared with four out of 12 control subjects in a previous series who received 1 ml of red cells from the same donor but without passively‐administered antibody. The time at which anti‐D was first detected and the maximum concentration of anti‐D attained were similar in test and control subjects. After a second injection of D‐positive red cells, given 5 months after the first, the total number of subjects making anti‐D became 11 out of 13 in the test series, compared with six out of 12 in the previous control series. Antibody concentrations were similar in test and control subjects. Evidently, Rh immunization was not augmented if this term implies accelerated formation of antibody or an obvious increase in antibody concentration. On the other hand, the suggestive increase in the proportion of responders, though not statistically significant, is consistent with the earlier observations of others and suggests that the effect of a small dose of passively‐administered antibody is to facilitate primary immunization.
T‐cell functional abnormality in B‐chronic lymphocytic leukaemia: evidence of a defect of the T‐helper subset
Authors: Lauria F. , Foa R. , Mantovani V. , Fierro M. T. , Catovsky D. , Tura S.
Published: July 2010

Summary. The helper and suppressor capacity of T, Tμ (T nonγ) and Tγ cells was assessed in a group of patients with B‐cell chronic lymphocytic leukaemia (B‐CLL) in a pokeweed mitogen (PWM) stimulated system. The enriched T‐cells (E‐rosette positive) from all B‐CLL cases showed a reduced capacity to induce the differentiation of normal B‐lymphocytes compared with normal T‐cells (P<0·005). After enrichment of the Tμ cells, the helper/inducer capacity was still significantly depressed compared with the same fraction from normal controls (P<0·01). On the other hand, enriched Tγ cells from B‐CLL were effective in suppressing the differentiation of normal B‐lymphocytes to a similar degree as normal Tγ cells. These findings are indicative of a deficient T‐cell helper function in B‐CLL, which appears to be unrelated to the clinical stage of the disease. The fractionation experiments suggest that this functional impairment is not only due to the abnormal T‐cell subset distribution seen in the majority of cases, but point to a possible intrinsic defect within the Tμ cell population.
Thalassaemia intermedia in Cyprus: the interaction of α and β thalassaemia
Authors: Wainscoat J. S. , Kanavakis E. , Wood W. G. , Letsky E. A. , Huehns E. R. , Marsh G. W. , Higgs D. R. , Clegg J. B. , Weatherall D. J.
Published: July 2010

Summary. Restriction endonuclease analysis has been performed on the α and β globin gene clusters of 57 Cypriots homozygous for β thalassaemia, 30 with the transfusion dependent form of the condition (thalassaemia major) and 27 who are less severely affected (thalassaemia intermedia). There was a significant difference in the incidence of α thalassaemia between the two groups: 14/27 of the patients with thalassaemia intermedia also had deletion forms of a thalassaemia, while only 4/30 of the patients with thalassaemia major were similarly affected. Thus in Cypriot patients who are homozygous for β thalassaemia the co‐inheritance of a thalassaemia is an important factor in determining the clinical course.
T cell regeneration after allogeneic and autologous bone marrow transplantation
Authors: Linch D. C. , Knott L. J. , Thomas R.M. , Harper P. , Goldstone A. H. , Davis E. G. , Levinski R.J.
Published: July 2010

Summary. Venous blood T cell phenotypes were analysed with monoclonal antibodies after 11 allogeneic and 17 autologous bone marrow transplants. In seven cases studied in the early regenerative period, cells with a thymocyte phenotype were present in the blood. In the large majority of patients treated with both allografts and autografts there was an imbalance of phenotypic ‘helper’ and ‘suppressor’ T cell subsets with initially a relative and later an absolute increase of ‘suppressor’ T cells. This imbalance was still present at over 250 d in eight out nine cases. Suppressor T cells bearing HLA‐Dr antigens were abundant in one case of fatal GVHD but not in another, and were also increased following two autografts. It is concluded that T cell phenotyping is not of diagnostic value in sick patients following bone marrow transplantation when graft‐versus‐host disease is suspected.
Liver disease in haemophiliacs: an overstated problem?
Authors: Stevens Richard F. , Cuthbert Ann C. , Perera Piyaseeli R. , Whitwell Helen L. , Haboubi Najib Y. , Warnes Thomas W. , Smith Alexander , Craske John , Longson Maurice , Wensley Richard T. , Delamore Irvine W.
Published: July 2010

Summary. Successful percutaneous liver biopsy was carried out on 12 multi‐transfused haemophiliacs from the Manchester area with persistently abnormal liver function tests. Only one patient showed evidence of chronic active hepatitis with progression to active micronodular cirrhosis although a further four patients showed some evidence of mild chronic active hepatitis. This represents a much lower incidence of severe histological liver damage than many previous reports and implies that liver biopsy in asymptomatic haemophiliacs may not be indicated as a routine procedure, particularly in the absence of proven therapy. Dynamic liver function tests may prove to be a useful indicator of deteriorating liver function in the otherwise asymptomatic haemophiliac.
Megakaryocyte colony formation by bone marrow progenitors in myelodysplasia syndromes
Authors: Juvonen E. , Partanen S. , Knuutila S. , Ruutu T.
Published: July 2010

Summary. Megakaryocytic colony formation by precursor cells from the bone marrow was investigated in 10 patients with a myelodysplastic syndrome. All but one exhibited abnormal colony formation: four showed no colony formation at all, while a decreased number of colonies was noticed in five. All of the patients showed defective colony formation by erythroid progenitors, but only four showed clearly abnormal granulocyte‐macrophage colony formation. The defect in megakaryocytic progenitors seems to be more akin to the defects occurring in erythroid progenitors than to the defects seen in the granulocyte‐macrophage lineage.
The relationship between chronic lymphocytic leukaemia and prolymphocytic leukaemia
Authors: Melo Junia V. , Catovsky Daniel , Galton David A. G.
Published: July 2010

Summary. The clinical and laboratory features of 300 patients with chronic lymphocytic leukaemia (CLL) and prolymphocytic leukaemia (PLL) of B‐cell type were studied in order to investigate the relationship between these two diseases. Statistical analysis demonstrated that more than 55% circulating prolymphocytes (PROL) was a defining criterion for PLL, disorder characterized by marked splenomegaly without lymph‐node enlargement, cells with high density of membrane‐immunoglobulin (Smlg), low mouse‐rosettes (M‐rosettes) and strong reactivity with the monoclonal antibody FMC7. Patients with typical CLL, defined as having less than 10% PROL, were on average 10 years younger than those with PLL and showed preferential lymph‐node to spleen involvement. Characteristic markers of CLL were weak Smlg, high M‐rosettes and low reactivity with FMC7. Patients with 11‐55% PROL, group designated as CLL/PL, were found to have intermediate features between CLL and PLL: the degree of splenomegaly was disproportionate to the lymph‐node enlargement, the number of cases with strong Smlg was closer to that found in PLL, but the other markers were not significantly different from CLL. The CLL/PL group appeared to be heterogeneous and includes at least two types of CLL, one with increased proportions of PROL but otherwise typical disease, and another in ‘prolymphocytoid’ transformation. Our study suggests that although PLL cannot be considered as the extreme end of a continuous spectrum from typical CLL, the spleen may be the source of PROL both in PLL and in CLL/PL.
No evidence for gamma‐interferon mediated haematopoietic inhibition by T cells in aplastic anaemia: an observation in the course of immunosuppressive therapy
Authors: Hanada Takashi , Yamamura Hideshi , Ehara Takao , Iwasaki Nobuaki , Shin Reiko , Nakahara Satoko , Takita Hitoshi
Published: July 2010

Two patients with aplastic anaemia were treated with immunosuppressive agents and peripheral blood T cells were cryopreserved serially. Inhibitory activity of T cells to autologous CFU‐E, γ‐IFN production by T cells and T cell subpopulations were assayed after remission. Inhibitory activity to autologous CFU‐E was not correlated with the numbers or ratios of T cell subpopulations. γ‐IFN production by T cells were within the normal range when inhibitory activity was found. In addition, γ‐IFN production increased after haematopoietic recovery. These findings suggest that γ‐IFN is not a soluble mediator of T cell‐mediated haematopoietic inhibition in aplastic anaemia.
Cloning of human erythroid progenitors (BFU‐E) in the absence of fetal bovine serum
Authors: Migliaccio Giovanni , Migliaccio Anna Rita
Published: July 2010

We describe culture conditions which enabled us to clone early human erythroid progenitors (BFU‐E) in the absence of fetal bovine serum (FBS). Our medium, which is chemically fully defined, supports proliferation and differentiation of erythroid progenitors comparable to that in FBS‐supplemented cultures. Furthermore, it allows cloning of BFU‐E from all human haemopoietic tissues (adult marrow, embryonic liver and yolk sac) and adult or perinatal blood. This system should facilitate the investigation of human erythroid differentiation in vitro (e.g. cell‐cell interaction. mechanism of action of haemopoietins and inhibitors, regulation of Hb synthesis) as well as the mechanisms underlying myeloproliferative disorders. As an example, we have found that recombinant Ep shows, in this culture system, the same dose response as Ep from conventional sources.
Androstane therapy to treat aplastic anaemia in adults: an uncontrolled pilot study
Authors: Gardner Frank H. , Juneja Harinder S.
Published: July 2010

During the past 9 years, 43 adult patients with severe aplastic anaemia have been treated with a combination of 3a etiocholanolone and nandrolone decanoate in an uncontrolled pilot study. Eleven patients were considered acute and the remainder as chronic severe aplasia. 50% (22 patients) had a haematopoietic recovery and 40% have had a sustained remission varying from 1‐5 to 8 years. Three patients who did not respond to 3a etiocholanolone had a haematologic reponse when treated with the isomer 3β etiocholanolone. The recovery did not appear to be associated with age or duration of marrow aplasia. In view of the reproducible chemical structure of these androstanes, we believe this group of steroids should be evaluated further in the treatment of severe aplastic anaemia.
α, β and γ T‐cell receptor genes: rearrangements correlate with haematological phenotype in T cell leukaemias
Authors: Foroni L. , Foldi J. , Matutes E. , Catovsky D. , O'Connor N. J. , Baer R. , Forster A. , Rabbitts T. H. , Luzzatto L.
Published: July 2010

We have studied the arrangement of the α, β and γT cell receptor (TCR) genes in 27 patients with T cell lymphoproliferative disorders. Nine patients had acute lymphoblastic leukaemia (T‐ALL), nine patients had prolympho‐cytic leukaemia (PLL), six patients presented with a T‐CLL/T‐lymphocytosis syndrome, two patients had Sezary syndrome (SS) and one patient had HTLV‐I positive T‐cell leukaemia/ lymphoma (ATLL). α TCR gene rearrangement could be demonstrated by the use of three available probes in only one case. By contrast, both β and γ TCR gene rearrangement could be demonstrated by Southern blot analysis of DNA samples digested with appropriate restriction enzymes in the majority of cases. In general, when rearrangements were present they involved both alleles. The proportion of rearranged chromosomes was lower in T‐ALL than in other forms of T‐cell leukaemia and it was lower in cases with the CD4 —/ CD8+ phenotype than in those with a CD4 +/CD8—phenotype. In three out of 34 cases of B‐cell leukaemia the TCR β‐gene but not the TCR γ‐gene was rearranged, just as in two out of 26 cases of T‐cell leukaemia the immunoglobulin (Ig) heavy chain but not the light chain genes were rearranged.

These data suggest that development of the machinery required for gene rearrangement may precede commitment to B or T cell lineage. The use of this technique is especially useful for the classification of cases of ALL in which the cells are negative with respect to most current phenotypic markers and in cases of T cell lymphocytosis in which the finding of a gene rearrangement identifies a monoclonal cell population.
A multivariate analysis of prognostic factors in chronic myelomonocytic leukaemia according to the FAB criteria
Authors: Ribera Josep‐Marǐa , Cervantes Francisco , Rozman Ciril
Published: July 2010

In an attempt to characterize the prognostic significance of the main initial clinical, haematological and biochemical features of chronic myelomonocytic leukaemia according to the FAB criteria, 29 such patients were analysed by means of univariate and multivariate statistical methods. At the time of the analysis 21 patients had died, with a median survival for the overall series of 8‐2 months. The univariate analysis identified three parameters associated with poor prognosis: high monocyte counts, low platelet counts and splenomegaly. When all the initial features were

included in a multiple regression model, only high monocyte counts and spleen enlargement retained their unfavourable prognostic influence (P=0‐002 and P = 002, respectively). Based on the presence or not of these prognostic factors, two populations of patients with different survival (median survival 5‐6and 16‐5months,respectively,P< 0‐005)could be identified. It seems therefore that monocytic proliferation serves not only as the diagnostic marker for chronic myelomonocytic leukaemia but also as the most important feature in the assessment of the patient's prognosis.
Room temperature ADP induced first stage hyperaggregation of human blood platelets: a previously undescribed phenomenon and its relationship to spontaneous cold induced platelet aggregation
Authors: Cohen Ian J. , Fuchs Jacob , Kaplinski Chaim , Krugliak Judith , Stark Batia , Vogel Ruth , Weinberger Itzhak , Rotenberg Zvi , Agmon Jacob , Jerushalmyi Zohara , Zaizov Rina
Published: July 2010

ADP induced human platelet aggregation was shown to be accentuated when tested at 20‐30°C as increased sensitivity and as a greater change of optical density although second stage aggregation and the release reaction did not occur. This previously undescribed phenomenon is defined as room temperature ADP induced first stage hyperaggregation. Aggregation, which occurs under the above mentioned conditions with a quantity of ADP insufficient to maintain the aggregation (usually less than 1 5 μm), is reversible when the temperature is raised to 3 7°C. After rewarming to these temperatures, second stage aggregation appeared in the presence of larger quantities of ADP (usually more than 2 μm) and could be blocked by aspirin. The absence of the release reaction was demonstrated with a lumi‐aggregometer. Spontaneous cold induced platelet aggregation seen after chilling platelets to 0‐4°C is shown to be a distinct phenomenon.
Phenotypic and genotypic analysis of chronic myelogenous leukaemia with T lymphoblastic and megakaryoblastic mixed crisis
Authors: Yasukawa Masaki , Iwamasa Kikue , Kawamura Shinichi , Murakami Satoshi , Takada Kiyonori , Hato Takaaki , Shiosaka Takahiko , Tamai Tomonori , Fukuoka Tamotsu , Fujita Shigeru , Kobayashi Yuzuru
Published: July 2010

A case of blast crisis in chronic myelogeneous leukaemia (CML) in which two distinct cell lineages were involved is presented. The phenotype of blasts in lymph nodes was T11 (CD2)+, Ia+, TdT+, suggesting T cell lineage. On the other hand, blasts in bone marrow and peripheral blood expressed platelet glycoprotein IIb/IIIa complex on their surface, suggesting megakaryocyte lineage. Cytogenetic analysis of lymph node and bone marrow cells revealed the abnormalities. inv(7) (p15q34) and t(1;3) (q23;q21), respectively, as well as the presence of the Ph1 chromosome in both cell types. Rearrangement of the T cell receptor β‐chain gene was detected in lymph node blasts, although blast cells in peripheral blood showed a germ line configuration. The involvement of T cell and megakaryocyte lineages in the blast crisis phase of CML was confirmed in our phenotypic and genotypic analysis, and the pathogenic association between blast crisis lineages and the additional chromosome abnormalities present is discussed.
Fibrinogen Aarhus and factor XIII induced polymerization and gel formation
Authors: Hessel Birgit , Adamson Lars , Procyk Roman , Therkildsen Lisbeth , Stenbjerg Stenar , Blombauck Birger
Published: July 2010

Fibrinogen Aarhus is an abnormal fibrinogen for which the clotting time with thrombin is greatly prolonged both in plasma and in the isolated fibrinogen. The whole blood clotting time is only slightly prolonged. The patient with this fibrinogen has no bleeding tendency. In this report we have investigated fibrinogen Aarhus in two alternative, thrombin independent polymerization and gelation pathways. These pathways are the factor XIII dependent oligomerization and gelation of fibrinogen, and heteropolymer (fibrinogen‐fibronectin) formation which also is catalysed by factor XIII. Both of these reactions are qualitatively the same in fibrinogen Aarhus as in normal fibrinogen, but the rate of oligomerization is somewhat slower in fibrinogen Aarhus. This may depend on impaired association between factor XIII and fibrinogen Aarhus.
Cefotetan‐induced immunologic thrombocytopenia
Authors: Christie Douglas J. , Lennon Shari S. , Drew Raymond L. , Swinehart Cheryl D.
Published: July 2010

Profound thrombocytopenia accompanied by a severe coagulopathy developed in an elderly female patient being treated with cefotetan while undergoing surgery for closure of a perforated gastric ulcer. During the acute phase of the bleeding diathesis, the patient had a platelet count of 12 × 109/1, a prothrombin time of 88 s (normal 10.0–11.8 s) and a PTT of 105 s (normal 23.0–37.0 s). Potent IgG cefotetan‐dependent anti‐platelet antibodies, which also were weakly reactive with ampicillin, were detected in the patient's serum using immunofluorescence and a recently developed protein A–agarose rosette forming assay. Unlike typical cephalosporin‐ and penicillin‐induced antibodies that react with cells pretreated with drug, this antibody only reacted with platelets in the presence of exogenous drug. Failure of the antibody to react with drug‐coated platelets suggests the possibility that, in this patient, sensitization to cefotetan involved mechanisms other than formation of typical hapten–carrier complexes normally described for members of the cephalosporin family of antibiotics. This appears to be the first definitive report that cefotetan, or any other cephalosporin derivative, can induce immunologic thrombocytopenia.
Megakaryocytic colony formation in polycythaemia vera and secondary erythrocytosis
Authors: Juvonen E. , Partanen S. , Ikkala E. , Ruutu T.
Published: July 2010

Megakaryocytic colony formation by progenitor cells of 18 patients with polycythaemia vera, seven with secondary erythrocytosis and four with erythrocytosis of unexplained origin was studied in vitro by the methyl cellulose culture assay. Fourteen of the 18 patients with polycythaemia vera showed spontaneous megakaryocytic colony formation, i.e. colony growth with normal human plasma as the only source of colony stimulation. None of the patients with secondary erythrocytosis or erythrocytosis of unknown origin or of the normal controls grew colonies in the presence of normal human plasma only. When the plasma of a patient with aplastic anaemia was used instead of normal human plasma and phytohaemagglutinin stimulated leucocyte conditioned medium (PHA‐LCM) was added to the culture medium, two of the patients with polycythaemia vera and one with secondary erythrocytosis formed slightly increased numbers of megakaryocytic colonies, while the rest of the patients showed normal colony formation. All of the patients with polycythaemia vera but none of those with secondary erythrocytosis or erythrocytosis of unknown origin showed spontaneous erythroid colony growth. The present study shows that most patients with polycythaemia vera form spontaneous megakaryocytic colonies in vitro. This phenomenon has recently also been demonstrated in essential thrombocythaemia and it is apparently analogous to spontaneous erythroid colony growth seen in all myeloproliferative disorders.
Determination of the growth fraction in monoclonal gammopathy with the monoclonal antibody Ki‐67
Authors: Lokhorst Henk M. , Boom Saskia E. , Terpstra Wim , Roholl Paul , Gerdes Johannes , Bast Bert J. E. G.
Published: July 2010

The monoclonal antibody Ki‐67 reacts with a nuclear antigen that is present only in proliferating cells. The proportion of Ki‐67 positive cells may therefore serve as a reliable measurement for the growth fraction in normal and neoplasmic cell populations. We have tested the significance of the MoAb Ki‐67 in the classification of monoclonal gammopathy and compared the results with the plasma cell labelling index. In benign monoclonal gammopathy the percentage of Ki‐67 positive plasma cells (median 1.6%) was significantly lower than in untreated multiple myeloma (median 9.6). Among the patients with more than 10% Ki‐67 positive plasma cells there were some very short survivors.

The largest growth fractions (median 41.8%) were found in patients with relapsing multiple myeloma indicating here a different growth pattern more resembling the high‐grade lymphomas. A linear correlation between the proportion of Ki‐67 positive plasma cells and the labelling index was not found. Determination of the plasma cell growth fraction with the monoclonal antibody Ki‐67 in monoclonal gammopathy may help to discriminate benign monoclonal gammopathy from multiple myeloma and will probably identify a subgroup of multiple myeloma patients with a poor prognosis, including those with relapsing multiple myeloma.
A reversible defect of platelet PDGF content in myeloproliferative disorders
Authors: Baglin T. P. , Price S. M. , Boughton B. J.
Published: July 2010

Platelet PDGF (platelet derived growth factor), platelet associated IgG and plasma levels of circulating immune complexes were measured in patients with chronic myeloproliferative disorders (primary myelofibrosis, primary proliferative polycythaemia and essential thrombocythaemia). Platelet PDGF was low in 11/12 patients, immune complexes were elevated in 11/16, and PlAIgG was elevated in all 14 patients in whom it was measured. There was no significant correlation between platelet PDGF and plasma levels of immune complexes (r= ‐0.5, P>0.1).

Treatment with busulphan and prednisolone for 2–3 months restored normal levels of platelet PDGF and suppressed plasma immune complex levels. Plasmapheresis lowered levels of immune complexes but had no effect on platelet PDGF. These results indicate that the low platelet PDGF levels in chronic myloproliferative disorders represent a reversible defect which is not directly related to the presence of immune complexes.
Primary thrombocythaemia: diagnostic criteria and a simple scoring system for positive diagnosis
Authors: Dudley J. M. , Messinezy M. , Eridani S. , Holland L. J. , Lawrie A. , Nunan T. O. , Sawyer B. , Savidge G. F. , Pearson T. C.
Published: July 2010

Summary Forty new patients with elevated platelet counts (>600 × 1O9/1) and without palpable splenomegaly were assigned to diagnostic groups defined by essentially conventional criteria after 3 months follow up: Proliferative (17), reactive (17) or unclassified (six). Mean platelet volume (MPV), platelet distribution width (PDW). platelet nucleotide ratio (ATP:ADP), unstimulated BFU‐E derived colony formation from peripheral blood. spleen scan and clinical ischaemia were assessed at the outset, with a view to defining diagnostic criteria for distinguishing primary thrombocythaemia from reactive thrombocytosis. All except the first variable were significantly associated with diagnostic group (P< 0.05). A simple scoring system was devised: enlarged spleen on scan, or presence of BFU‐E, each scored 2: elevated PDW (> 2 SD from mean), elevated ATP:ADP (>4 SD from mean) or presence of clinical ischaemia, each scored 1. Score totals ± 3 predicted primary thrombocythaemia, and totals < 3 suggested reactive thrombocytosis (predictive value 89%). The system correctly predicted diagnosis in four out of four (probably six out of six) patients whose diagnosis was not apparent initially, and thus whose results were not used in constructing the scoring system. Exclusion of BFU‐E from the system resulted in only one incorrect prediction in this group.
GIMEMA ALL 0183: a multicentric study on adult acute Iymphoblastic lecukaemia in Italy
Authors: Mandelli F.
Published: July 2010

SummaryIn 1983 a multicentre trial for adult patients with ALL (excluding the B‐phenotype) was initi,ated to evaluate: (a) complete remission (CR) rate and length in a protocol with a mild induction and an intensive consolidation therapy; (b) the efficacy of a mild versus an intensive post consolidation treatment in prolonging disease‐free survival; (c) CNS relapse rate on a protocol substituting intrathecal chemotherapy and intermediate dose i.v. methotrexate (MTX) and cytosine arabinoside (ARA‐C) for cranial radiotherapy. 358 patients entered the protocol between January 1983 and April 1987: 284 (79.3%) attained complete remission, 26 (7.3%) died during induction and 48 (13.4%) had resistant disease. Median overall survival was 21.7 months, with a projection that 29.4% of the patients will survive more than 55 months. The median duration of complete remission was 19.2 months: 21.9% of responders are projected to remain in continuing complete response for more than 50.6 months. Among the 284 responding patients, 154 (54.2%) relapsed (95 while on therapy, 59 off‐therapy): major sites of relapse were bone marrow (103). CNS alone (23), or both (11). 14 patients died while in CR. Only 12 patients (2.2%) were lost to follow‐up, while therapy was interrupted in 7 (2%) because of side effects and in 12 (3.3%) because of the patient's refusal. By multivariate analysis increasing age and presence of infection at diagnosis were associated with a lower remission rate: risk of relapse was greater in LL subtypes and in patients with higher WBC at diagnosis; survival was inversely correlated to age. The high response rate was not achieved at expense of an increased risk of induction death. The low incidence of isolated CNS relapse (8.0% of responding patients) suggests that CNS prophylaxis as used in this study is effective. No statistically significant difference in survival and relapse rate emerged between the two arms of post consolidation therapy. Moreover, the protocol therapy was well tolerated, required minimal supportive care, which could be provided even in small centres, and compliance proved acceptance for patients and staff.
Two novel polyadenylation mutations leading to β+‐thalassaemia
Authors: Jankovic L. , Efremov G. D. , Petkov G. , Kattamis C. , George E. , Yang K‐G. , Stoming T. A. , Huisman T. H. J.
Published: July 2010

In an ongoing effort to identify point mutations causing β‐thalassaemia, we have found two previously unreported mutations which are located in the Poly A site of the β‐globin gene. The screening programme used amplified DNA and dot‐blot hybridization with several 32P‐labelled oligonucleotide probes. DNA samples which remained unidentified by this methodology were subjected to sequencing with 32P‐labelled primers and modified T7 DNA polymerase. The newly discovered mutations were confirmed by the dot‐blot hybridization technique. One type concerned an AATAAA→ÁTGAA mutation in the polyadenylation site and was found in one family from Yugoslavia (including one patient with the C→T mutation at codon 29 in trans), one from Bulgaria (the patient had the G→A mutation at IVS‐I‐110 in trans), and one from Greece (this patient had the C→G mutation at IVS‐II‐745 in trans). Haematological data for three simple heterozygotes suggested a rather mild β+‐thalassaemia. The second type involved an AATAAA→ÁTAGA mutation and was found in one family from Malaysia. The propositus had the βE mutation on the other chromosome, was originally diagnosed as mild Hb E‐β+‐thalassaemia, and had Hb A and Hb E percentages which were nearly the same.
Characterization of polymorphic markers in the von Willebrand factor gene and pseudogene
Authors: Bernardi F. , Marchetti G. , Casonato A. , Gemmati D. , Patracchini P. , Legnani C. , DeRosa V. , Girolami A. , Conconi F.
Published: July 2010

Three TaqI restriction fragment length polymorphisms (RFLP) detected by the central portion of von Willebrand factor cDNA, which recognizes the true gene and in addition pseudogenic sequences, were characterized and mapped. Small cDNA fragments which hybridized with DNA from families with von Willebrand disease were used.

Two of the RFLP, recognized by 1.7 and 0.45 kb cDNA fragments, are not in linkage either with von Willebrand disease or with RFLP located in the von Willebrand factor (vWF) gene, which indicates their pseudogenic location. These markers located in 22q11, near to the bcr gene, provide new tools for the study of several somatic and constitutional alterations affecting this chromosomal region.

The third RFLP is recognized by a cDNA fragment corresponding to the N‐terminal portion of mature vWF and is localized in the true gene. Since significant linkage disequilibrium with other informative RFLP is not present, this marker contributes to the definition of family haplotypes associated with von Willebrand disease.