bluuebunny/crossref_metadata_2025 huggingface dataset:
Persistent Epstein‐Barr virus infection associated with monosomy 7 or chromosome 3 abnormality in childhood myeloproliferative disorders
Authors: Stollmann B. , Fonatsch Ch. , Havers W.
Published: March 2008

Summary

This report deals with myeloproliferative disorders associated with chronic, persistent Epstein‐Barr virus (EBV) infection and with monosomy 7 and aberrations concerning chromosomes 3 and 5. Altogether five children were affected, their age ranging from 1 to 4 years at time of clinical diagnosis. Principal symptoms were: hepatomegaly, splenomegaly, recurring upper respiratory tract infection and anaemia. The serum IgG level remained persistently increased.

Anti EBV antibody concentrations were measured over a period of 9 months to 6 years, demonstrating persistently increased concentrations of IgG antibodies to viral capsid antigen (VCA) and against early antigen (EA). In three patients IgA antibodies were also studied and were found to be elevated.

Within 2‐5 years two children developed chronic myelomonocytic leukaemia from the chronic myeloproliferative syndrome. A third patient who initially was diagnosed as chronic myelomonocytic leukaemia developed acute leukaemia within a period of 12 months. A fourth patient with myeloproliferative syndrome went into spontaneous remission after an observation period of 2 years. A fifth patient, the only one with translocation t(3;5)(q27;q33), displayed symptoms and a clinical course that can best be characterized as juvenile chronic myelocytic leukaemia. The clinical, haematological, serological and cytogenetic findings may be related.
Book Reviews
Authors: Galton D. A. G. , Marcus Robert
Published: March 2008

Book Reviews in This Article:

Human Leukemias: Cytochemical and Ultrastructural Techniques in Diagnosis and Research. Developments in Oncology Series. Edited by A. Polliack.

Leukaemia Reviews International. Edited by Marvin A. Rich.
The gene triplet Rw W Ph controls murine haematopoiesis
Authors: Loutit J. F. , Cattanach B. M. , Lyon Mary F.
Published: March 2008

Summary

Having shown a haematopoietic role for Patch (Ph), especially when doubly heterozygous with the mutant Wct (Loutit & Cattanach, 1983) we have similarly examined the Rw locus, the third member of the triplet. Mature Rw/+ mice have a just detectable macrocytic anaemia. When doubly heterozygous with Wct and Wv the mild anaemia of these W mutants is exaggerated but with W19H (not anaemic as single heterozygote) red cell factors are as for Rw/+. Rw/+ mice are strikingly more sensitive to the lethal effects of X‐irradiation (MLD 6.66 ± 0.10 Gy) on haemotopoiesis than comparable +/+ mice (MLD 8 Gy). Those mice that do recover after X‐irradiation do not exhibit the delay in recovery of erythropoiesis that is evident with characteristic W mutants in both single and double dose. Furthermore the double heterozygote Wct+/+Rw has a significantly lower MLD (5.19 ± 0.17 Gy) for X‐radiation than Wct/+ (MLD 6.48 ± 0.24 Gy). We argue that all three loci W, Ph and Rw, influence haematopoietic stem cells, leading to increased radiosensitivity when deletions or mutant genes are present.
Cytogenetic studies on recipients of allogeneic bone marrow transplants after fractionated total body irradiation
Authors: Schmitz N. , Gödde‐Salz Elisabeth , Löffler H.
Published: March 2008

Summary

Cytogenetic findings from the bone marrow (BM) and the peripheral blood (PB) of nine consecutive patients after allogeneic bone marrow transplantation (BMT) for acute or chronic myelogenous leukaemia are reported. After a conditioning regimen consisting of cyclophosphamide and fractionated total body irradiation (TBI) given in five or six fractions of 2 Gy, persistence of host cells was detected in four out of seven cases with permanent engraftment. While one of these patients relapsed 4 months after host cells had been found in BM and PB, the other patients stayed relapse‐free 124, 257 and 347 d after grafting. Before transplantation, the leukaemic cells in all three cases carried unique cytogenetic abnormalities giving us the opportunity to distinguish the leukaemic population from chromosomally non‐aberrant cells thought to represent residual normal host cells. As the persisting host cells after BMT lacked any cytogenetic abnormalities, it is suggested that they were members of residual normal clones not involved in the leukaemic process.
Lack of correlation between nucleated bone marrow cell dose, marrow CFU‐GM dose or marrow CFU‐E dose and the rate of HLA‐identical sibling marrow engraftment
Authors: Atkinson Kerry , Norrie Susan , Chan Pik , Downs Kate , Biggs James
Published: March 2008

Summary

There was no correlation between the rate of marrow engraftment and the number of nucleated bone marrow cells infused into 50 HLA‐identical sibling marrow graft recipients with haematological malignancy conditioned with cyclo‐phosphamide and fractionated total body irradiation, and immunosuppressed with either cyclosporin (42 patients) or methotrexate (eight patients). Similarly, there was no correlation between the number of marrow CFU‐GM or CFU‐e infused into recipients immunosuppressed with cyclosporin. The data show that recipients of HLA‐identical sibling marrow allografts conditioned with cyclophosphamide and total body irradiation require less than 3 × 108 nucleated bone marrow cells/kg recipient weight to ensure engraftment, and throw doubt on the relevance of measuring committed progenitor cells in the donor marrow to assess the likelihood of subsequent haemopoietic reconstitution after matched sibling transplantation.
Kinetics, in vivo redistribution and sites of sequestration of indium‐111‐labelled platelets in giant platelet syndromes
Authors: Heyns Anthon du P. , Badenhorst Philip N. , Wessels Paula , Pieters Henry , Lötter Mattheus G.
Published: March 2008

Summary

Six patients with giant platelet syndrome were examined: four with Bernard‐Soulier syndrome (two were asplenic); one with hereditary thrombopathic thrombocytopenia; and one with May‐Hegglin anomaly. Autologous platelets were labelled with In‐111‐oxine and in vivo redistribution and sites of sequestration measured with quantitative imaging. In Bernard‐Soulier syndrome platelet survival was normal or moderately shortened; platelet turnover was decreased only in the two patients with thrombocytopenia. In the patients with thrombopathia or May‐Hegglin anomaly, platelet survival and turnover was moderately decreased. In those patients with normal‐sized spleens, the mean splenic platelet pool consisted of 35.5% of the platelet mass, i.e. normal. The intrasplenic transmit time of the megathrombocytes was prolonged. Splenic blood flow was within normal limits. There was a marked accumulation of platelets in the liver at equilibrium: 15.5‐58.8% of whole body radioactivity (normal 9.6±1.2%). This finding is unexplained. The final sites of sequestration of platelets were mainly in the liver and spleen, similar to that seen in normal subjects. We conclude that there is no inverse relationship between cell size and splenic platelet transit time. Platelet size therefore does not determine the size of the splenic platelet pool. The size of the platelets also does not seem to affect the sites of sequestration at the end of their life span.
Platelet aggregation occurs in congenital afibrinogenaemia despite the absence of fibrinogen or its fragments in plasma and platelets, as demonstrated by immunoenzymology
Authors: Soria Jeannette , Soria Claudine , Borg Jeanne Yvonne , Mirshahi Massoud , Piguet Hubert , Tron Philippe , Fessard Claude , Caen Jacques Philipe
Published: March 2008

Summary

Platelet aggregation of an afibrinogenaemic patient's platelet rich plasma (PRP) was greatly decreased when ADP was used for stimulation. In the presence of collagen or arachidonic acid the changes in light transmission recorded during platelet aggregation of patient's PRP were similar to those observed with normal PRP but the size of aggregates appeared to be smaller in comparison with those observed with normal platelets. In addition, thrombin‐induced aggregation of washed platelets was similar to normal platelets. The interpretation was made possible because the fibrinogen level in plasma and in platelets was found to be almost nil as demonstrated by both an Elisa procedure described here and the determination of fibrinopeptide A (fpA). Furthermore, fibrinogen fragments, which could result from abnormal synthesis and therefore replace fibrinogen in platelet aggregation, were undetectable by immunological analysis using specific antibodies against Aα, Bß and γ chains and 10 different monoclonal antibodies against fibrin degradation products.
Antibodies to human adult T cell leukaemia virus type I associated antigens in Swedish leukaemia patients and blood donors
Authors: Blomberg Jonas , Fäldt Roger
Published: March 2008

Summary

Antibodies to antigens associated with human T cell leukaemia virus type I (HTLV I) in Swedish adult leukaemia patients and blood donors were sought with a sensitive screening test using membrane antigen prepared from virus producing cells (MA‐ELISA). Four persons (one ALL, one AML and two healthy blood donors) out of 483 persons tested reacted in the test. However, they were negative in the more specific anti‐p19 and anti‐whole virion ELISA tests. The prevalence of sera with definite anti‐HTLV I activity seems to be very low in Sweden. The finding of four MA‐ELISA positive persons needs further investigation.
Cyclosporine and graft failure following bone marrow transplantation for severe aplastic anaemia
Authors: Hows J. , P almer S. , Gordon‐Smith E. C.
Published: March 2008

Summary

Thirty‐nine patients with severe aplastic anaemia were transplanted from HLA identical sibling donors. Irradiation was not used in the pre‐transplant immunosuppressive protocol and cyclosporine was used as the post graft immuno‐suppressive agent. The incidence of primary graft failure (no take) was low occurring in 3/38 evaluable patients. Late graft failure 4–7.5 months post BMT occurred in five patients and was associated with withdrawal of cyclosporine therapy. Mixed lymphocyte chimaerism was demonstrated in 3/5 cases at the time of late graft failure and subsequent marrow recovery was autologous in four patients. Mortality from graft failure was low with 2/38 evaluable patients (5%) dying from this complication.
Cumulative antiplatelet effect of low‐dose enteric coated aspirin
Authors: Jakubowski Joseph A. , Stampfer Meir J. , V aillancourt Raymond , Deykin Daniel
Published: March 2008

Summary

Enteric coated aspirin (ECA) at doses of 325–1300 mg is an effective alternative to regular aspirin for inhibition of platelet activity while avoiding gastric irritation. The objectives of this study were to determine: (1) the lowest chronic dose of ECA providing effective inhibition of platelet activities, (2) the time course of the inhibition, and (3) the reappearance of platelet cyclo‐oxygenase activity. Seven subjects were studied before and after seven daily doses of 40–325 mg ECA. Serum thromboxane (TX) B2 levels indicated that the lowest dose of ECA resulting in greater than 90% inhibition of platelet cyclo‐oxygenase was 80 mg/d. Platelet aggregation and ATP release in response to collagen (1 μg/ml) and arachidonic acid (1 mM) were abolished and bleeding times were prolonged from 6.1 ± 1.5 min to 9.7 ± 2.8 min (mean ± SD, P<0.01). Examination of platelet cyclo‐oxygenase activity on a daily basis revealed that 24 h after the first 80 mg dose serum TXB2 had decreased by approximately 60% and was suppressed by more than 90% after four doses. Recovery of platelet cyclo‐oxygenase activity after a single 80 mg dose of ECA was delayed for 48–72 h indicating that aspirin reached the systemic circulation. We conclude that chronic inhibition of platelet activity may be achieved in a cumulative manner with 80 mg ECA/d.
Regulation of human peripheral blood BFU‐E growth in vitro by leukaemic B‐lymphocytes
Authors: Najman Albert , Baillou Claude , Drouet Xavier , Leblanc Ginette , Douay Luc , Claude Gorin Norbert , Duhamel Gearard
Published: March 2008

Summary

We report a stimulating effect of leukaemic B‐lymphocytes from anaemic and non‐anaemic patients with CLL on the proliferation of normal peripheral blood BFU‐E. Coculture of leukaemic B‐cells at various concentrations (2.5 x 103‐106) with 2.5 x 105 mononuclear cells from normal peripheral blood increased the number of BFU‐E derived erythroid colonies. The same effect was observed when the number of target cells was varied in the presence of a fixed number of B‐lymphocytes, with a clear linear relationship. B‐cell conditioned medium gave a similar increase when added to the culture instead of B‐cells. At high concentration of B‐cells from anaemic patients, the size of the colonies was increased and a large number of macroscopic colonies was seen. The place of the B‐cells in the regulation of erythroid progenitors in relation to monocytes and T‐lymphocytes has still to be established.
Carrier detection in factor VII congenital deficiency
Authors: Mariani G. , Hermans J. , Orlando M. , Mazzucconi M. G. , Ciavarella N. , Faiella A. , Berrettini M. , Chistolini A. , Di Nucci G. D. , Mannucci P. M. , Mandelli F.
Published: March 2008

Summary

Thirty obligate and 28 possible carriers of factor VII congenital deficiency, belonging to 16 families, were studied in relation to the immunological variants to which the kindreds belonged, namely, VII+, VIIR and VII‐. Factor VII activity and antigen determinations in these subjects formed two phenotypical patterns: a discrepant pattern characterized by a low ratio activity/antigen present in VII+ heterozygotes, and a non‐discrepant pattern (normal ratio activity/antigen) which is present in the VII‐ and VIIR variants.

In the first genetic variant the detection of carriers can be performed using the ratio VII:C/VII:Ag. In the other variant, which accounts for the vast majority of heterozygotes, the distribution of the carriers’factor VII is so widespread that a large overlap results between these subjects and the normals. The application of a probabilistic calculation performed by combining the actual values of factor VII: C and the genetic probability of carriership using Fisher's linear discriminant analysis, makes discrimination between carriers and normals easier.
Ex vivo T‐cell depletion with the monoclonal antibody Campath‐1 plus human complement effectively prevents acute graft‐versus‐host disease in allogeneic bone marrow transplantation
Authors: Heit W. , Bunjes D. , Wiesneth M. , Schmeiser T. , Arnold R. , Hale G. , Waldmann H. , Heimpel H.
Published: March 2008

Summary. We have developed a rapid and simple procedure for the elimination of mature T‐cells from the donor marrow using a single incubation with the monoclonal antibody Campath‐1 and donor complement. This resulted in a reduction of T‐cell contamination to a mean of 1%. This regimen reduced the incidence of acute graft‐versus‐host disease significantly in 21 consecutive bone marrow grafts in 18 patients with leukaemia and non‐Hodgkin's lymphoma. Purging was responsible for an increased incidence of graft rejection in HLA‐identical transplants (13%).
Cytogenetic evidence for involvement of erythroid progenitors in a child with therapy linked myelodysplasia
Authors: Grier H. E. , Weinstein H. J. , Révész T. , Houlihan P. W. , Greenhalgh C. L. , Nathan D. G. , Tantravahi R.
Published: March 2008

Summary. A 6‐year‐old male with prior metastatic retinoblastoma developed a therapy linked myelodysplastic syndrome. Whole bone marrow cytogenetics showed monosomy 7 and a marker chromosome. To determine the progenitor level of origin of the malignant clone, we studied the karyotypes of marrow erythroid and granulocyte/macrophage colonies grown in methyl cellulose. All erythroid and granulocyte/macrophage colonies had an abnormal karyotype with 45 chromosomes (monosomy 7) and several colonies contained the marker chromosome. These findings give direct evidence that this patient's myelodysplastic syndrome involved an early stem cell which was capable of both erythroid and granulocyte differentiation.
Surface marker analysis in acute myeloid leukaemia and correlation with FAB classification
Authors: Miguel J. F. San , Gonzalez M. , Cañizo M. C. , Anta J. P. , Zola H. , Borrasca A. Lopez
Published: March 2008

Summary. The immunological phenotype of blast cells in 102 patients with acute myeloid leukaemia (AML) was analysed with a panel of 20 monoclonal antibodies and the enzyme terminal transferase, and correlated with the FAB classification. Although a partial correlation between these two approaches could be observed, almost every morphological group contained patients from more than one immunological phenotype. The M1 and M5a leukaemias showed the most undifferentiated phenotype, often lacking in specific myelomo‐nocytic antigens. The M3 formed a uniform group defined as My7+, Ia‐, FMC8+, a phenotype which was also observed in two cases of the microgranu‐lar variant. The granulocytic (CDwl5) and monocytic (CDwl4) antibodies crossreacted with some M5b and M2 leukaemias, respectively. Compared with M5a, the M5b leukaemias showed a large increase in the expression of CDw l4 antigen, confirming the validity of the morphological differentiation. Giyco‐phorin‐A was present in four out of five M6 leukaemias. TdT activity was demonstrated in 10% of AML cases, with a higher incidence among the monocytic variants: M4 and M5‐. Eleven AML were considered as unclassifi‐able according to the FAB criteria and in seven of them a megakaryoblastic cell population (GP IIb/IIIa+, GPIb+) was demonstrated; this confirms the need to include the subgroup of megakaryoblastic leukaemias within the AML. Finally, a possible immunological classification for AML is proposed.
Variant translocation t(8;22) and abnormalities of chromosome 15(q22) and 17(q 12–21) in a Burkitt's lymphoma/leukaemia with disseminated intravascular coagulation
Authors: Daly Peter , Brito‐Babapulle Vasantha , Lawlor Emer , Blaney Claire , Parreira Antonio , Catovsky Daniel
Published: March 2008

Summary. Severe disseminated intravascular coagulation was observed in a patient with Burkitt's lymphoma/leukaemia. Immunological studies on leukae‐mic blasts from relapsed bone marrow revealed a B‐cell phenotype (B4+. B1+, HLA‐ Dr+, J5+) with membrane bound IgMλ Cytogenetic investigation revealed a variant Burkitt's translocation t(8;22)(q24;q11) involving the λ light chain gene region and abnormalities of chromosomes 15 and 17 with breakpoints at q22 and q 12 respectively, similar to those observed in the t(15:17) in acute promyelocytic leukaemia. Transmission electron microscopy of the leukaemic blasts showed crystalline cytoplasmic inclusions which may have had a role in precipitating the disseminated intravascular coagulation.
Binding of thrombin‐activated human factor VIII to platelets
Authors: Soberano Mercedes E. , Clarke Denise , Zucker Marjorie B.
Published: March 2008

Summary. To study association of platelets with factor VIII, the purified protein was 125I‐labelled with Bolton‐Hunter reagent to a specific activity of 243 000–360 000 cpm/U. Autoradiographs of SDS polyacrylamide gels revealed polypep‐tides of VIII at Mr about 240 kDa, 90 kDa and intermediate values, as well as some radioactive contaminants, but the light chain (Mr 78/76) seen with silver stain was not labelled. After 2.5–5‐fold activation with thrombin, the higher radioactive Mr band disappeared, the band at 90 kDa became more intense, and a band appeared at about 45 kDa. The radioactivity associated with platelets, studied in the presence of haemophilic BaS04‐treated plasma, was maximal after 6–8 min and increased 3–15‐fold on activation with thrombin. With activated VIII, autoradiographs of platelet pellets showed only Villa but results are expressed as units of unactivated VIII bound. At 0.3–0.7 U/ml, 108 platelets bound 0.0008–0.004 U Villa. The amount bound was not affected by the ratio of unlabelled VIII to VIII labelled in the presence of a 50‐fold molar excess of unlabelled Bolton‐Hunter reagent. Binding increased to 1.5 U Villa/108 platelets (about 13 600 molecules per platelet) at 140 U/ml, with no evidence of saturation. Binding was not affected by monoclonal antibodies to platelet glycoproteins IIb/IIIa or Ib, quenching the thrombin before adding platelets, or aggregating the platelets with A2318 7 in the presence of thrombin. Qualitatively, binding of labelled Villa and factor Va studied by others are similar. Binding of 125I‐Villa to aggregated and unaggregated platelets was normal in patient M.S. whose platelets were shown by others to be deficient in their ability to bind radiolabelled factor Xa and generate coagulant activity. This difference, and the fact that platelet coagulant activity is increased by platelet activation and/or aggregation, suggest that binding of Va or Villa alone does not determine the assembly of active proteolytic complexes on the platelet surface.
Erythropoiesis in pregnancy
Authors: Howells M. R. , Jones S. E. , Napier J. A. F. , Saunders K. , Cavill I.
Published: March 2008

Summary. The reticulocyte count, serum ferritin and serum erythropoietin concentrations were measured during the course of pregnancy in 41 women. A decrease in iron stores early in pregnancy was accompanied by a significant rise in the reticulocyte count until the 28th week. Erythropoietin levels did not rise significantly until the 28th week of pregnancy and had fallen to the post‐natal level by 40 weeks. The results are consistent with an increase in erythropoietic activity early in pregnancy which did not appear to depend upon increased plasma erythropoietin levels.
Induction of colony‐stimulating factor response in myeloid leukaemia cell lines
Authors: Rhyner Kaspar , Taetle Raymond
Published: March 2008

Summary. Untreated, late passage HL60 promyelocytic and KG1 myeloblastic leukaemia cells did not increase proliferation with placenta or Mo T cell conditioned medium containing colony‐stimulating factor (CSF) nor with partially purified, recombinant granulocyte/macrophage (GM)‐CSF. However, after induction with DMSO or 1,25‐dihydroxy‐vitamin D3, HL60 cells showed dose‐dependent increases in proliferation with crude and purified CSFs. CSF responses and macrophage differentiation were induced in KG1 cells by treatment with tetradecanoylphorbol‐acetate (TPA). When cells were exposed to inducing agents for varying periods, washed and exposed to CSF, proliferative responses were related to time of exposure. Cells exposed for 1–4 d showed post‐induction CSF‐induced proliferation, but cells induced for 5–6 d were inhibited by CSF. Induction of CSF response appeared linked to differentiation, since KG1 cells differentiated with TPA and developed CSF‐induced proliferative responses, but showed no differentiation or CSF induced proliferation after treatment with vitamin D3. When HL60 cells were continuously exposed to DMSO or vitamin D3, overall cell production was increased by placenta conditioned medium, but cultures still became senescent and died after several weeks. Cells continuously cultured with DMSO were predominantly macrophages, indicating lineages of DMSO‐induced differentiation were modified by continuous culture or the presence of CSF. After treatment with chemical inducers, proliferation of myeloid leukaemia lines is stimulated by CSF, providing a model for post‐deterministic regulation of normal and malignant myeloid cell production.
Myeloid and erythroid lineage expression of haemopoietic progenitors derived from an abnormal clone in erythroleukaemia
Authors: Suda Toshio , Sato Yuko , Furukawa Yusuke , Suda Junko , Eguchi Mituoki , Saito Masaki , Miura Yasusada
Published: March 2008

Summary. To clarify the lineage involvement of haemopoietic progenitor cells in erythroleukaemia, the morphology and chromosomes of single colonies from a patient with erythroleukaemia were analysed simultaneously. The cytogenetic analysis of bone marrow cells revealed two clones; 44.XY, −7, −12, −17, del (5)(q31),+ Mar and 43,XY, −7, −12, −17, −19,del(5)(q31),+ Mar. Of 40 metaphases examined, there were 34 and six of these clones, respectively. Bone marrow mononuclear cells were plated at 5 × 104/ml in methylcellulose medium containing phytohaemagglutinin‐stimulated leucocyte conditioned medium and erythropoietin. Seventeen colonies, i.e. nine blast cell colonies, four myeloid (Sudan black B‐positive) colonies, and four erythroid (benzidine‐positive) colonies contained analysable metaphases, yielding 102 metaphases in total. Except for chromosome random loss, the karyotype within a colony remained constant. All three types of colonies showed an abnormal clone; 44.XY, − 7, −12, − 17,del(5)(q31),+Mar. From these findings, it is concluded that myeloid and erythroid lineages in erythroleukaemia were derived from the same abnormal clone.
Characterization of platelet‐specific alloantigens by immimoblotting: localization of Zw and Bak antigens
Authors: van der Schoot C. Ellen , Wester Mieke , von dem Borne Albert E. G. Kr. , Huisman Han G.
Published: March 2008

Summary. The glycoprotein localization of the platelet‐specific antigens Zwa, Zwb and Baka and their presence on tryptic fragments of glycoproteins was studied by immunoblotting. Human platelets were solubilized and pre‐cleared from platelet‐associated IgG. The glycoproteins were separated on SDS polyacrylamide gels, transferred to nitrocellulose and incubated with platelet antibodies, followed by 125I‐radiolabelled anti‐human Ig antibodies. Glycoprotein Ilb/IIIa were isolated from platelet lysates by immuno‐affinity chromato‐graphy. These proteins were subjected to trypsin digestion, and then used for the immunoblot procedure with platelet antibodies. A glycoprotein specifically reacting with either anti‐Zwa or anti‐Zwb was found, with an apparent molecular weight of 88 kDa. This protein co‐migrated, and was probably identical with, glycoprotein IIIa. After trypsin digestion the smallest fragment, reactive with IgG anti‐Zwa or IgM anti‐Zwb, had a molecular weight of approximately 23 kDa. IgG anti‐Baka and anti‐Leka antibodies reacted with a protein of 130 kDa from platelets of Bak(a+) donors. This protein was identified as glycoprotein IIb.
The incidence, clonal origin and secretory nature of serum paraproteins in chronic lymphocytic leukaemia
Authors: Sinclair D. , Dagg J. H. , Dewar A. E. , Mowat A. Md. , Parrott D. M. V. , Stockdill G. , Stott D. I.
Published: March 2008

Summary. Immuno‐isoelectric focusing (IIEF) showed a 61% incidence of serum paraproteinaemia in 56 patients with chronic lymphocytic leukaemia (CLL). A strong correlation between the serum paraprotein heavy chain isotypes and those of the cytoplasmic heavy chain immunoglobulins was observed with no discrepancy noted in light chain expression. Density gradient ultracentrifuga‐tion analysis of selected sera containing monoclonal IgM showed that the IgM paraproteins were mostly 19S, secretory IgM but one patient was found to have both 19S and 8S monoclonal IgM. When the cellular origin of the IgM and IgD paraproteins found in one patient was investigated, both paraproteins were found to share the same idiotype and originate from the neoplastic clone.

These findings confirm the view that there is an incomplete maturation block in chronic lymphocytic leukaemia and that in vivo secretion of paraproteins by the neoplastic cells is a relatively common occurrence.
The incidence of monoclonal gammopathy in a population over 45 years old determined by isoelectric focusing
Authors: Sinclair D. , Sheehan T. , Parrott D. M. V. , Stott D. I.
Published: March 2008

Summary. Immuno‐isoelectric focusing (IIEF), a technique previously shown to be sensitive for the detection of paraproteinaemia, was used to test 200 individuals over the age of 45, without history of B cell neoplasm, for the presence of serum paraproteinaemia. 11% of these individuals had evidence of paraproteinaemia detectable by IIEF compared with only 2% by zonal and immunoelectrophoresis. A further 12% had oligoclonal immunoglobulins and the remainder had no qualitative abnormality of the immunoglobulin profile. These results are discussed with particular reference to the aetiology, diagnosis and monitoring of potential B cell neoplasm in high risk individuals or groups.
In vitro growth of sheep erythroid progenitors
Authors: Kelly Susan J. , Wood W. G.
Published: March 2008

Summary. We have attempted to define culture conditions suitable to support the in vitro growth of early erythroid progenitors (BFU‐E), derived from the liver or bone marrow and blood of fetal and adult sheep. Testing a wide variety of conditions showed that only some batches of fetal lamb sera made a notable difference to growth. BFU‐E from normal fetal blood were demonstrated sporadically; they appeared regularly in low numbers in blood taken during erythroid (anaemic) stress. Adult blood only contained sporadic BFU‐E during severe anaemic stress. This difficulty in demonstrating circulating BFU‐E in sheep may be due to a genuine paucity of such cells or to an undefined requirement for in vitro growth.
Certain myeloid cells possess receptors for interleukin‐2
Authors: Armitage R. J. , Lai A. P. , Roberts P. J. , Cawley J. C.
Published: March 2008

Summary. A proportion of blasts from five of 10 cases of AML expressed receptors for IL‐2 (IL‐2R) when tested directly ex‐vivo with monoclonal antibodies against the receptor. After in‐vitro stimulation with various agents including TPA, γ interferon and colony stimulating factor, the purified blast cells of all cases of AML tested (10 of 10) showed high levels (50–90% cells positive) of IL‐2R expression.

Granulocytic cells from the promyelocyte stage onwards lacked IL‐2R both before and after in‐vitro stimulation. In contrast, leukaemic promonocytes and normal peripheral monocytes expressed IL‐2R both before and after stimulation.
Proposed serum standard for human serum vitamin B12 assay
Authors:   International Committee for Standardization in Haematology (ICSH)
Published: March 2008

The International Committee for Standardization in Haematology (ICSH) expert panel on vitamin B12 assay has undertaken a multi‐laboratory study to develop a vitamin B12 serum standard.
Red cell associated IgG in patients suffering from Plasmodium falciparum malaria
Authors: Abdalla S. H.
Published: March 2008

Summary. Quantitation of red cell associated IgG in 62 Gambian patients with P. falciparum malaria and 23 normal adult controls was carried out, using a purified 125I‐labelled anti IgG. The number of IgG molecules per red cell was found to be between 90–897 molecules for patients with malaria and 100–233 for controls. 12 patients with malaria had raised levels of RBC associated IgG. There was no correlation between severity of anaemia and RBC associated IgG levels in patients with malaria nor was there a correlation between reticulocytosis and RBC associated IgG levels. It is concluded that although immune haemolysis may occur in some patients with malaria who have high levels of IgG or activated complement components on their red cells, other factors such as marrow suppression or ineffective erythropoiesis play an important role in the pathogenesis of the post‐malaria anaemia.
Iron uptake by MOLT 3 cells from transferrin/monoclonal antitransferrin antibody complexes
Authors: Hradilek Antonín , Neuwirt Jan
Published: March 2008

Summary. The ability of seven monoclonal antibodies (MAbs) reactive with human transferrin (Tf) to inhibit iron uptake from Tf by cells of the human lymphoid line MOLT 3 was compared with the effect of these MAbs on Tf binding to MOLT 3 surface Tf receptors. MAbs HTF‐01, HTF‐06, HTF‐07, HTF‐11 and HTF‐14 inhibited both iron uptake and Tf binding. Complexing Tf with MAbs HTF‐04 or HTF‐05 resulted in an increased association of Tf with the cells but iron uptake was diminished. Following the intracellular kinetics of Tf/HTF‐04 complex has shown that the whole complex is endocytosed and Tf iron is retained inside the cell, but Tf release from both the cell surface and the intracellular compartment is slower when compared to Tf alone. Iron uptake inhibition was therefore attributed to lengthening of the Tf cell cycle after complexing of Tf with HTF‐04 and it is suggested that the rate of cellular Tf turnover might, together with the number of functioning Tf receptors, determine the rate of iron uptake by cells.
Acute megakaryoblastic leukaemia: a prospective study of its identification and treatment
Authors: Ruiz‐Argüelles G. J. , Marín‐Lopez A. , Lobato‐Mendizábal E. , Ruiz‐Argüelles A. , Nichols W. L. , Katzman J. A.
Published: March 2008

Summary. Acute megakaryoblastic leukaemia remains an uncommonly recognized disorder, usually identified by electron microscopy or with monoclonal antibodies. Using two monoclonal antibodies (HP1‐1D identifying glycoprotein IIB/IIIA and W1‐23 identifying von Willebrand factor) we have studied during a 12 month period all cases of acute leukaemia with FAB L2, M1 or undifferentiated morphology presenting in our cooperative group ‘Grupo de Hematólogos de Puebla’ in México. Six of 21 FAB L2, M1 or undifferentiated leukaemias were classified immunologically as megakaryoblastic. Five of these patients had myelofibrosis and two had normal platelet counts but abnormal platelet aggregation function. All six patients were treated with low‐dose cytosine arabinoside (10 mg/m2) administered subcutaneously, twice daily in 21 d courses). Haematologic response was achieved in five cases while the sixth died before completing one course. Three patients relapsed within the first month after completing the chemotherapy and died. Two patients remain in remission 11 and 15 months after initial treatment. It is our impression that the prevalence of acute megakaryoblastic leukaemia has been under‐estimated, and that low‐dose cytosine arabinoside treatment may be of value in its management.
Ferritins in malignant and non‐malignant lymphoid cells
Authors: Vezzoni Paolo , Levi Sonia , Gabri Elena , Pozzi Maria Rosa , Spinazze Silvia , Arosio Paolo
Published: March 2008

Summary. Lymphoid cells from peripheral blood, thymus, malignant and non‐malignant lymph nodes were analysed for ferritin content using radioimmunoassays specific for the ‘acidic’ H‐subunit‐rich and for ‘basic’ L‐subunit‐rich isoferritins, and the data were compared with the immunological characteristics of the cells. All tissues with high proportion of T or ‘null’ cells contained the lowest concentration of L‐subunit‐rich isoferritins, while the H‐subunit‐rich forms increased from low levels in the quiescent peripheral blood lymphocytes (PBL), to higher values in the immature and proliferating thymocytes and lymphoblasts, malignant or not. B‐cell lymphomas contained concentrations of both ferritin types higher than those found in PBL. No significant difference was found in the isoferritin concentrations between non‐malignant lymph nodes and tissues involved in Hodgkin's disease. These findings indicate that maturation stage, proliferative status and anatomical localization affect isoferritin expression in lymphoid cells.
Evidence against a platelet cyclooxygenase defect in uraemic subjects on chronic haemodialysis
Authors: Bloom A. , Greaves M. , Preston F. E. , Brown C. B.
Published: March 2008

Summary. Defective platelet thromboxane synthesis has been described in uraemia and attributed to a ‘functional cyclooxygenase defect’. We have studied platelet aggregation and generation of immunoreactive thromboxane B2 (TXB2) in 11 subjects on a chronic haemodialysis programme. The platelet function abnormality of uraemia was confirmed, maximal aggregation in response to collagen (2 and 4 μg/ml) and sodium arachidonate (1·5 and 3·0 mm) being significantly depressed. However, increased platelet aggregation in response to sodium arachidonate 0·75 mM was noted. Due to the reduced haematocrit, the platelet concentration in platelet‐rich plasma (PRP) of uraemic subjects was significantly lower than that of controls; when TXB2 generation in PRP adjusted to 200 × 109 platelets/l was assessed, no evidence for a defect of cyclooxygenase was found, although reduced synthesis of TXB2 in response to thrombin was noted. Furthermore, increased thromboxane generation by uraemic PRP in response to sodium arachidonate 0·75 mM was detected.

We conclude that the mild platelet abnormality in uraemic subjects treated by haemodialysis is not explained by a ‘functional cyclooxygenase defect’, although an abnormality of thrombin‐induced thromboxane synthesis may be present. Furthermore, the tendency to increased aggregation and thromboxane synthesis in response to a low concentration of arachidonic acid may contribute to the thrombotic tendency which is also described in such subjects.
Transferrin modulation of colony stimulating factor elaboration by adherent blood and bone marrow cells
Authors: Taetle Raymond , Honeysett J. Michael
Published: March 2008

Summary. When added to cultured normal, adherent blood and marrow cells at concentrations of 25–100μg/ml (3 × 10−7to 1·3 × 10−6m), human transferrin enhanced colony‐stimulating factor (CSF) elaboration. Fe saturated and relatively unsaturated Tf were equally effective in increasing CSF release, but soluble ferric nitriloacetatc was ineffective. Dose‐dependent increases in CSF release by adherent blood and marrow cells occurred in serum‐free media and the continuous presence of Tf was necessary for this effect. Using a monoclonal anti‐Tf receptor antibody, normal blood mononuclear cells contained no detectable Tf receptor positive cells. However, after 5 d culture in serum‐free medium, Tf receptor positive cells were identified among normal adherent blood cells, and the per cent receptor positive cells increased with Tf concentration. We conclude Tf modulates CSF production from normal, adherent blood and marrow cells in vitro. These findings indicate a possible role for Tf in intramedullary regulation of normal granulopoiesis.
Primary thrombocythaemia treated with busulphan
Authors: Van de Pette J. E. W. , Prochazka A. V. , Pearson T. C. , Singh A. K. , Dickson E. R. , Wetherley‐Mein G.
Published: March 2008

Summary.Thirty‐seven patients with primary thrombocythaemia (PT) treated with busulphan have been followed for periods up to 25 years. Reduction of the platelet count to less than 400 × 109/l resolved vascular occlusive symptoms, but haemorrhagic symptoms often remained unaltered. Cox regression analysis indicated that there were only two prognostically important presenting features; age had a strong inverse correlation with survival and vascular occlusive symptoms correlated with a better survival. Median duration ofsurvival on treatment was 9·8 years. The number of deaths was 2·1 times that of a comparable control group, with deaths from myelofibrosis markedly increased. Deaths from thrombosis and malignant diseases, including leukaemia, were not significantly different from the number expected. which emphasizes the efficacy and the relative safety of busulphan for the long‐term treatment of PT. Progression of PT into myelofibrosis occurred in 24% of cases and 9% became polycythaemic. Two additional cases of ‘thrombocythaemia'with a Philadelphia chromosome (and no overt evidence of chronic granulocytic leukaemia) are also presented.
DNA content and cell cycle analysis of bone marrow cells in myelodysplastic syndromes (MDS)
Authors: Peters S. W. , Clark R. E. , Hoy T. G. , Jacobs A.
Published: March 2008

Summary. DNA distribution in bone marrow cells from 10 normal subjects and 34 patients with MDS (24 with refractory anaemia (RA) and 10 with excess blasts (RAEB)) was determined by flow cytometry using a FACS III. DNA histograms were resolved into GO/1, S and G2/M compartments by fitting Gaussian distributions and the DNA content of GO/1 cells, expressed as DNA index (DI), was determined. In 19 MDS patients the DI was outside the normal range, those with RA tending to be hyperdiploid. Two patients with RAEB had a second GO/1 peak. In five cases of RA the number of cells in GO/1 was below the normal range and the mean value was significantly lower than normal for this group as a whole (P= 0·018), Eleven (RA) patients had a higher percentage of cells in G2/M than normal (P= 0·001). In RAEB patients there is a suggestion of increased numbers in GO/1 and a decrease in S phase and G2/M cells. All three of the deaths amongst RA patients and four out of the five deaths in RAEB patients since the beginning of this study were associated with an abnormal DI at the initial investigation.
Evaluation of a new enzyme‐linked immunoassay for terminal deoxynucleotidyl transferase in haematologic malignancies
Authors: Sue Coleman Mary , Hui Ahn Yong , Fairbanks Tod , Manderino† George , Cibull Michael
Published: March 2008

Summary. We have evaluated a new solid phase enzyme immunoassay (EIA) for detection of terminal deoxynucleotidyl transferase (TdT). The EIA is greater than 100 times more sensitive than previously used tests for enzyme activity. In 284 clinical specimens of human peripheral blood and bone marrow, the EIA detected TdT antigen in 97% of peripheral blood and 100% of bone marrow samples that were positive for enzymatic activity. The excellent sensitivity and specificity of this new test suggests that it can be used in clinical situations where quantitative TdT measurements are desired.
The natural anti‐α‐galactosyl IgG on human normal senescent red blood cells
Authors: Galili Uri , Flechner Iancu , Knyszynski Ahuva , Danon David , Rachmilewitz Eliezer A.
Published: March 2008

Summary. A highly sensitive antiglobulin test based on rosette formation due to the interaction between IgG bearing red blood cells (RBC) and Fc receptors on K562 cells, was used to study the immunoglobulin molecules present on human senescent RBC. Normal human RBC were separated into young and senescent subpopulations on the basis of age‐dependent differences in density by centrifugation on a discontinuous density Percoll gradient, and by flotation on phthalate ester mixtures. The senescent but not the young RBC were found to bear membrane bound IgG. Most of the bound IgG molecules could be specifically eluted by galactose in its α‐anomeric form. Antigalactosyl (anti‐Gal) IgG antibodies with similar reactivity were found to be present in high titres in every one of the 400 normal human sera tested. The natural anti‐Gal antibodies isolated from normal sera by affinity chromatography could bind to IgG depleted senescent RBC but not to young RBC. Erythrophagocytosis experiments indicated that the anti‐Gal bound to the senescent RBC induced their destruction by macrophages. It is suggested that the natural anti‐Gal antibodies interact with cryptic α‐galactosyl residues which are exposed in the course of the RBC senescence and mediate the removal of these RBC from circulation by cells of the reticuloendothelial system.
Transitory hypomegakaryocytic thrombocytopenia: aetiological association with ethanol abuse and implications regarding regulation of human megakaryocytopoiesis
Authors: Gewirtz Alan M. , Hoffman Ronald
Published: March 2008

Summary. We studied a patient with a long history of ethanol abuse who presented to the hospital with profound weakness, anaemia and thrombocytopenia. Evaluation of these problems revealed the patient's bone marrow to be hypercellular but severely iron depleted and almost totally devoid of morphologically recognizable megakaryocytes. However, we were able to detect the presence of non‐morphologically recognizable, immature megakaryocytes in the same sample using an immunochemical detection technique. This circumstance allowed us to study the relative importance of both megakaryocyte maturation and peripheral blood platelet count on the production of megakaryocyte colony stimulating activity (Meg‐CSA), a putative regulator of the megakaryocyte colony forming unit (CFU‐M). The results of our investigations disclosed a rapid decline in serum Meg‐CSA levels which preceded recovery of the platelet count and appeared to coincide with the maturation of megakaryocytes into the morphologically recognizable pool. The effect of ETOH on the patient's CFU‐M cloning efficiency was also studied. ETOH in amounts up to 454 mg/dl did not inhibit cloning of the patient's peripheral blood CFU‐M in plasma clot cultures. Our results suggest that regulation of Meg‐CSA production is a complex function which appears to be dependent on a number of factors including the level of megakaryocyte maturation in the marrow. We also speculate that ethanol associated thrombocytopenia may occasionally be brought about by a disruption in the process of megakaryocyte maturation at the level of a progenitor more mature than the CFU‐M.
The activity state of factor VII in plasma: two pathways for the cold promoted activation of factor VII
Authors: Muller A. D. , van Deijk W. A. , Devilée P. P. , van Dam‐Mieras M. C. E. , Hemker H. C.
Published: March 2008

Summary. The apparent amount of factor VII as determined in a one‐stage test depends on the type of thromboplastin used: bovine thromboplastin only reacts with human factor VIIa whereas human thromboplastin interacts with unactivated human factor VII as well. Therefore the ratio factor VII activity as measured with bovine thromboplastin divided by the factor VII activity as assessed with human thromboplastin reflects the state of activation of factor VII in plasma. This approach was used to study the process of cold promoted factor VII activation and the involvement of different clotting factors therein. It could be shown that cold promoted activation does not occur in the absence of factors II and XII and is reduced for about 50% in factor IX deficient plasma. The other coagulation factors have a minor influence on the process. The results indicate that the cold promoted factor VII activation is the result of activation by both activated contact products and thrombin.
An association between the factor VII coagulant activity and thrombin activity induced by surface/cold exposure of normal human plasma
Authors: Miller G. J. , Seghatchian M. J. , Walter S. J. , Howarth D. J. , Thompson S. G. , Esnouf M. P. , Meade T. W.
Published: March 2008

Summary. To test whether factor VII activation correlated with the generation of thrombin activity when plasma was exposed to a glass surface and reduced temperature, an association was sought between the changes in factor VII clotting activity (VIIc) and fibrinopeptide A concentration (an index of thrombin activity) in platelet‐poor citrated plasma from 42 healthy adults. The Spearman rank correlation (rs) between responses was 0.82 (P < 0.001). The VIIc assay response to surface/cold exposure was unaffected when thrombin was suppressed by hirudin. An assay for factor VII activity based upon its activation of tritiated factor X revealed an association between the increase in fibrinopeptide A concentration and reduction in functional factor VII concentration during activation of a subset of 22 plasma samples (rs= ‐0.62; (P < 0.003). This loss of functional factor VII was probably due to conversion of active factor VII to its non‐functional end‐product by factor Xa. The results suggest that VIIc is an index of flux within the coagulation system and support the hypothesis that a high VIIc is an indicator of a hypercoagulable state.
The relation of exposure time and drug concentration in their effect on cloning efficiency after incubation of human bone marrow with cytosine arabinoside
Authors: Raijmakers R. , de Witte T. , Linssen P. , Wessels J. , Haanen C.
Published: March 2008

Summary. The effect of different cytosine arabinoside (Ara‐C) concentrations and exposure times upon clonogenicity of normal bone marrow cells was studied. One hour incubation, even at the pharmacologically high Ara‐C concentration of 10−5 M, was not cytotoxic. Prolonged incubation resulted in a very great increase in Ara‐C cytotoxicity, provided that inactivation of Ara‐C by deamination was limited. The results show that long‐term exposure to Ara‐C, even at pharmacologically very low concentrations, is deleterious to clonogenic cells and provide an experimental basis for the reported therapeutic efficacy of long‐term low dose Ara‐C administration in leukaemia and preleukaemic syndromes.