bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sickle cell‐β+ thalassaemia in Orissa State, India
Authors: Kulozik Andreas E. , Bail Sigrid , Kar Bimal C. , Serjeant Beryl E. , Serjeant Graham E. 
Published: March 2008

SummaryThe clinical, haematological, and some molecular genetic features of 17 Orissan Indian patients with sickle cell‐β+ thalassaemia (Sβ+ that) are described and compared with those in 131 Indian patients with homozygous sickle cell (SS) disease. Patients with Sβ+ that had higher Hb A2 levels, and lower mean cell volume (MCV) and mean cell haemoglobin (MCH) compared to SS disease but no other haematological difference of statistical significance. High levels of Hb F occurred in both genotypes and the α+ thalassaemia gene frequency reached 0 · 47 in Sβ+ thai and 0 · 32 in SS disease. Clinically there were no significant differences between the genotypes indicating that the low levels of HbA (3‐5%) in this condition were insufficient to modify the clinical features.The thalassaemic β globin gene is inactivated by a G → C mutation at position 5 of the first intron of the β globin gene (IVS1–5 G → C) in all cases. This finding should facilitate the introduction of a prenatal diagnosis programme aimed at the prevention of β thalassaemia or Sβ+ thalassaemia in that population.
Productive infection of in vitro generated haemopoietic progenitor cells from normal human adult peripheral blood with parvovirus B19: studies by morphology, immunocytochemistry, flow‐cytometry and DNA‐hybridization
Authors: Serke Stefan , Schwarz Tino Francis , Baurmann Herrad , Kirsch Andreas , Hottenträger Barbara , Brunn Albrecht Von , Roggendorf Michael , Huhn Dieter , Deinhardt Friedrich 
Published: March 2008

Parvovirus B19 exerts a highly selective cytopathic effect on erythroid progenitor cells. Studies so far on the pathogenesis of B19‐infection have been performed using bone marrow samples providing large amounts of erythroid progenitor cells. Extensive study, however, has been hampered by the limited access to bone marrow samples. We have designed a liquid culture method allowing the generation of large numbers of erythroid progenitor cells, initiating cultures with CD3‐ and CD14‐poor peripheral blood mononuclear cells. Following a 12 d preincubation in liquid cultures containing recombinant human interleukin 3 (rhll‐3) and recombinant human erythropoietin (rhEpo), cells harvested from the liquid cultures were exposed to B19‐containing plasma, followed by a further cultivation in liquid culture for up to 96 h. Cells expressing the CD13 and the glycophorin A (GlyA) antigens, respectively, were monitored sequentially by flow‐cytometry, demonstrating a selective inhibition of GlyA‐positive cells following B19‐inoculation. Typical morphological changes were observed on cytocentrifuge‐spots, and typical giant‐cells were identified as staining for GlyA. Productive infection by B19 was demonstrable, as B19‐DNA increased by about x 100 after 72 h of culture.The liquid culture method generating erythroid target cells for effective infection by B19 virus promises to be a useful and easily accessible tool for further research on B19 infection of haemopoietic cells.
A common Vδ2—Dδ2—Dδ3 T cell receptor gene rearrangement in precursor B acute lymphoblastic leukaemia
Authors: Yano Takahiro , Pullman Amy , Andrade Rafael , Uppenkamp Michael , de Villartay Jean Pierre , Reaman Gregory , Crush‐Stanton Sylvia , Cohen David I. , Raffeld Mark , Cossman Jeffrey 
Published: March 2008

Despite their apparent commitment to the B lymphocytic lineage, human precursor B cell acute lymphoblastic leukaemias (ALL) frequently rearrange their T cell antigen receptor (TCR) α, β and γ chain genes. Since these three genes are active sites of rearrangement in precursor B cell neoplasms, it seemed that the recently discovered fourth TCR gene, δ, might be similarly rearranged. To investigate this possibility, a series of precursor B cell leukaemias was analysed for rearrangements at the δ chain gene locus, using probes of the variable, joining, and constant regions of the δ chain gene.The majority of precursor B cell ALLs in this series (25/32, 78%) showed rearrangement or deletion of one or more TCR δ genes. This contrasts sharply with a series of 16 mature B cell neoplasms (chronic lymphocytic leukaemia) in which no TCR δ gene rearrangements were detected.An unusual TCR δ rearrangement, rarely observed in normal or neoplastic T cells, was seen in the majority (14/18) of precursor B cell ALLs with TCR δ rearrangements. In contrast to the utilization ov Vδ1 in T cell ALL, detailed restriction mapping of precursor B ALL revealed an incomplete rearrangement without involvement of Jδ segments. Direct genomic sequencing was performed on one example and demonstrated a nonproductive Vδ2‐Dδ2‐Dδ1 recombination in this precursor B ALL.We conclude that the TCR δ chain gene is an active locus in precursor B cell neoplasia, involves an unusual type of rearrangement and provides a clonal tumour marker for diagnosis of precursor B ALL.
Antithrombin Glasgow II: alanine 382 to threonine mutation in the serpin P12 position, resulting in a substrate reaction with thrombin
Authors: Ireland H. , Lane D. A. , Thompson E. , Walker I. D. , Blench I. , Morris H. R. , Freyssinet J. M. , Grunebaum L. , Olds R. , Thein S. L. 
Published: March 2008

A female with recurrent thrombosis was found to have a functional abnormality of antithrombin, with a ratio of functional to immunological activity in plasma of ∼ 50%. Crossed immunoelectrophoresis in the presence of heparin was normal, indicating an abnormality of the reactive site, rather than the heparin binding domain. Accordingly, the antithrombin was isolated by heparin‐Sepharose chromatography: this produced a mixture of normal and variant antithrombin, as the patient was heterozygous for the abnormality. To remove the normal component, the antithrombin was passed through a column of thrombin‐Sepharose. On sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE), prior to its application to thrombin‐Sepharose, the antithrombin migrated as a single band with identical mobility to that of normal antithrombin. After thrombin‐Sepharose, the purified variant component was proteolysed, and migrated as two components, one with a reduced and one with enhanced mobility under non‐reducing conditions. This demonstrated that the variant was unable to form stable inhibitor‐thrombin complexes and was cleaved in a substrate reaction with thrombin. One site of cleavage was unambiguously ascertained to be the Arg 393‐ Ser 394 reactive site bond, by NH2 terminal sequencing of the cleaved variant antithrombin: 10 steps beginning at the P 1’position, Ser‐Leu‐Asn‐Pro‐Asn‐Arg…., were clearly identified.The mutation responsible for this defect was studied by polymerase chain reaction (PCR) amplification of exon 6 of the antithrombin gene and direct sequencing of the amplified product. The presence of both a G and A in the first position of codon 382, identified the mutation GCA to ACA, which results in the substitution of Ala 382 to Thr. This is identical to that reported for antithrombin Hamilton (Devraj‐Kizuk et al, 1988), although antithrombin gene polymorphism analysis suggests that the antithrombin Glasgow II mutation has arisen independently.We have recently shown (Caso et al, 1991) that mutation at a nearby position, Ala 384 to Pro, also transforms another variant, antithrombin Vicenza/Charleville, into a substrate for thrombin. The present results with antithrombin Glasgow II suggest that all the alanine residues at the base of the reactive site loop in positions P12–10 may be important for the formation of a stabilized inhibitor‐thrombin complex.
Role of spleen in hereditary spherocytosis: evidence for increased in vitro proteolysis of red cell membrane
Authors: De Matteis M. C. , Angelis V. De , Sorrentino F. , Bonollo E. , Vettore L. 
Published: March 2008

In vitro proteolysis of red cell membranes has been studied by means of electrophoretic separation on SDS‐polyacrylamide gel of solubilized ghost proteins and subsequent densitometry of separated, stained bands; the amounts of major membrane proteins were measured in ghosts either with inhibited or with allowed proteolysis in the following cases: 15 patients suffering from hereditary spherocytosis (HS) with variable degree of spleen enlargement, eight cirrhotic patients with spleen enlargement and 12 healthy blood donors as control group.Proteolysis was present to a greater extent in HS patients with larger splenomegaly, lesser in HS with smaller splenomegaly, and was comparable to healthy controls both in splenectomized HS and in patients with spleen enlargement due to liver cirrhosis.The results suggest the involvement of splenomegaly in the enhancement of in vitro proteolysis in HS red cell membrane; it is probably attributable to joint effects of the damage induced in red cells by prolonged retention within haemolysing spleen together with the abnormalities genetically affecting the structure of HS red cell membrane.
The treatment of patients with newly diagnosed poor prognosis acute myelogenous leukaemia: response to treatment and treatment failure
Authors: Preisler Harvey D. , Larson Richard A. , Raza Azra , Browman George , Goldberg Jack , Vogler Ralph , Day Roger , Gottlieb Arlan , Vardiman James W. , Bennett John , Kukla Cathi , Grunwald Hans 
Published: March 2008

The failure of poor prognosis patients with newly diagnosed AML to enter remission is usually due to two phenomena: a high mortality rate and resistance of the leukaemia to chemotherapy. We conducted a pilot study of a regimen designed to overcome these two types of treatment failure. Patients were carefully selected for therapy on the basis of their likelihood of surviving. Chemotherapy consisted of high dose cytosine arabinoside (HDaraC) with the doses modified on the basis of patient age so as to reduce the risk of toxicity. Finally, daunorubicin was administered only to those patients for whom HDaraC was not likely to produce sufficient antileukaemia effects to produce a remission.The median patient age was 67 years and 69/88 (78%) patients had a history of preleukaemia and/or toxic exposure. Only 11/84 (13%) patients died during remission induction therapy and 40/84 (48%) entered CR. Patients with only one risk factor had a higher CR rate than those with more than one risk factor (56%v. 30%, P= 0·02) and also had longer durations (222 d v. 113·5 d, P= 0·035). Two types of resistance to chemotherapy were observed: ‘classical resistance’(the failure of chemotherapy to produce substantial killing of leukaemia cells) and the rapid regrowth of leukaemia cells subsequent to a level of cytoreduction which otherwise would have been sufficient to produce a CR.
Abnormal low and high density lipoproteins in homozygous beta‐thalassaemia
Authors: Goldfarb Ada W. , Rachmilewitz Eliezer A. , Eisenberg Shlomo 
Published: March 2008

The levels, structure and composition of plasma lipoproteins were determined in 67 patients with homozygous beta‐thalassaemia and compared to healthy or heterozygous members of the same families and to patients with either sickle cell or iron deficiency anaemia. Plasma total and LDL and HDL cholesterol levels were low in patients with homozygous beta‐thalassaemia and with sickle cell anaemia. Plasma triglycerides did not differ between subjects. The low plasma and lipoprotein cholesterol was independent of age, transfusion requirements and splenectomy.Abnormal structure and composition of lipoproteins was found in homozygous beta‐thalassaemia. The LDL was of higher density and was triglyceride‐rich and cholesterol ester‐poor. HDL separated to three populations. HDL2was prominent (in spite of low plasma HDL cholesterol). HDL2–3was of normal density and an intermediate HDL population, not found in normal subjects, was identified and designated HDL2–3. All three HDL populations were enriched with triglycerides and poor in cholesterol ester content. The modified LDL and HDL particles may then be possibly cleared rapidly from the plasma by activated monocytes and macro‐phages.
Clonality of cell populations in refractory anaemia using combined approach of gene loss and X‐linked restriction fragment length polymorphism‐methylation analyses
Authors: Abrahamson Gail , Boultwood Jacqueline , Madden Jacqueline , Kelly Susan , Oscier D. G. , Rack Katrina , Buckle Veronica J. , Wainscoat J. S. 
Published: March 2008

We have used X‐linked restriction fragment length polymorphism (RFLP)‐methylation and gene deletion analyses to investigate the nature of the progenitor cell of origin in the myelodysplastic syndromes (MDS). Gene deletion studies were performed on the granulocyte and T‐lymphocyte fractions of six women with refractory anaemia (RA) and either a partial deletion of the long arm of chromosome 5 (5q‐) or monosomy 7. All six showed gene loss in the granulocyte but not the T‐lymphocyte fractions, indicating monoclonality of the granulocytes but not the T‐lymphocytes. In order to further investigate this finding, we subsequently performed X‐RFLP‐methylation studies using the probe M27β, and also a probe for the phosphoglycerate kinase (PGK) gene. These studies have confirmed the monoclonality of the granulocytes and the polyclonality of the T‐lymphocytes in these cases. Our findings suggest that in this group of patients with MDS the T‐lymphocytes were not involved in the disorder, and furthermore, in the one case where B‐lymphocytes were also available, that the progenitor cell of origin was restricted to the myeloid lineage.This work was supported by the Leukaemia Research Fund of the United Kingdom. We thank Dr I. Craig and Professor A. Jeffreys for the probes M27β and pγ3, respectively. The probe for the FMS gene was kindly donated by Dr N. Spurr. The probes for the HPRT and PGK gene regions were used with the permission of Dr B. Vogelstein. We thank Dr D. Y. Mason for use of the monoclonal antibodies CD22 and CD68.
Second transplants for leukaemic relapse after bone marrow transplantation: high early mortality but favourable effect of chronic GVHD on continued remission. A REPORT BY THE EBMT LEUKAEMIA WORKING PARTY
Authors: Barrett A. J. , Locatelli F. , Treleaven J. G. , Gratwohl A. , Szydlo R. , Zwaan F. E. 
Published: March 2008

Ninety second bone marrow transplants (BMT) for relapsed leukaemia were carried out in 30 European BMT centres. At second BMT, after further treatment in 64 cases, 43 patients were in complete remission or in chronic phase of CML, and 47 were in continuing relapse, accelerated phase or blast crisis of CML. Seventy patients died, 37 from early transplant‐related toxicity and relapse or failure to eradicate leukaemia which occurred in 23. There were 20 survivors. The actuarial disease‐free survival was 11% with a relapse probability of 69% at 3 years. Associated with reduced graftversus‐host disease (GVHD) prophylaxis during second BMT, the incidence and severity of acute and chronic GVHD, was increased when compared with the first BMT (P= 0.02, and 0.002 respectively for acute and chronic GVHD).In multivariate analysis survival was shown to be favoured by a prolonged interval between first and second BMT (relative risk 1.3/year,P= 0.02), and no or mild chronic GVHD following first BMT (relative risk 2.3,P= 0.02). Continuing remission was favoured by chronic GVHD occurring after second BMT (relative risk 8.1,P= 0.004).These results confirm the high treatment‐related mortality following second BMT, but identify superior survival in selected patients. Improved results might be achieved by further reduction in preparative regimen intensity, and increasing graft‐versus‐leukaemia reactivity.
In vitro Ig‐synthesis and proliferative activity in multiple myeloma are stimulated by different growth factors
Authors: Sonneveld Pieter , Schoester Martijn , Leeuw Kees de 
Published: March 2008

Plasma cells isolated from bone marrow (BM) aspirates of 15 patients with active multiple myeloma (MM) were cultured and analysed for in vitro proliferative response and Ig‐synthesis upon stimulation with interleukin‐3 (IL‐3), interleukin‐4 (IL‐4) and interleukin‐6 (IL‐6). The proliferative response, determined as Ki‐67 positivity in concentrated plasma cells, was increased by IL‐6 (Stimulation Index, SI=1.77±0.21 (M±SEM)) but not by IL‐3 or IL‐4. This proliferation could be blocked by anti‐IL‐6. In vitro Igsynthesis was stimulated by IL‐4 (SI = 1.62±0.12 (M±SEM) P<0.05) but not by IL‐6 or IL‐3. This effect was not antagonized by anti‐IL‐6. An inverse correlation was found in this group of patients between the IL‐6 induced stimulation of plasma cell proliferative activity and the IL‐4 induced increase of Ig‐synthesis (P=0.027).These data indicate in MM that Ig‐synthesis and the in vitro proliferative activity may be stimulated by different haematopoietic growth factors, which may reflect the involvement of different responding cells.
Treatment of multiple myeloma with interferon alpha: the Scandinavian experience
Authors: Mellstedt H. , ÖSterborg A. , Björkholm M. , Björeman M. , Brenning G. , Gahrton G. , Grimfors G. , Gyllenhammar H. , Hast R. , Juliusson G. , Jaurnmark M. , Killander A. , Kimby E. , Lerner R. , Merk K. , Ohrling M. , Simonsson B. , Smedmyr B. , Stalfelt A. M. , Strander H. , Svedmyr E. , Udean A. M. , Wadman D. , ÖSby E. 
Published: March 2008

IFNα is a biologic therapeutic agent with documented antitumoural effect in multiple myeloma. 15% of previously untreated myeloma patients achieve a clinical response to IFNα alone with a possible dose‐response relationship. A particularly good effect is noted in IgA myelomas treated with natural IFNα.A randomized study was started in April 1986 comparing melphalan/prednisone (MP) therapy with MP plus natural IFNα (MP/IFN) in untreated patients with multiple myeloma stages II and III. 220 patients had entered the study by autumn 1989. An interim report is given here. The response frequency was 48% in the MP group and 66% in the MP/IFN group (P<0·02). Stage II patients responded better to MP/IFN (76%) than to MP alone (48%) (P<0·01). No significant difference was noted for stage III patients. 91% of all IgA myelomas responded to MP/IFN and 52% to MP (P<0·01). The difference in response frequency of IgG and BJ myelomas between the two treatment groups was not statistically significant. The observation period is still too short to draw firm conclusions on survival. However, a statistically significant longer response duration time and survival from response (P<0·01) was noted for stage II patients.
Immunomagnetic bead isolation of megakaryocytes from guinea‐pig bone marrow: effect of recombinant interleukin‐6 on size, ploidy and cytoplasmic fragmentation
Authors: Leven Robert M. , Rodriguez Adrian 
Published: March 2008

SummaryGuinea‐pig bone marrow megakaryocytes were isolated using an antibody to platelet glycoprotein Ib and a second antibody conjugated to magnetic beads. The procedure yielded an average of 644 800 megakaryocytes from two guinea‐pigs with an average viability of 83%. All of the platelet glycoprotein Ib positive cells also expressed the platelet glycoprotein IIb–IIIa complex. The size and ploidy of megakaryocytes isolated by this technique were analysed in the presence of 10 ng/ml of interleukin‐6 (IL‐6). Without IL‐6 megakaryocyte size increased significantly after 24 h. but an even larger increase in size occurred in the presence of IL‐6. The modal ploidy class was 16N with an average of 19% 2N. 2·6% 4N, 16·4% 8N. 50·8% 16N and 11·1% 32N cells as determined by flow cytometry. Measurements made by microspectrophotometry were in close agreement. After 24 h incubation there was a significant rise in the percentage of 2N and 32N cells. The ploidy distribution after 24 h with IL‐6 was the same as the control. Megakaryocytes cultured in the absence of serum on collagen gels did not form pseudopods and fragment, as occurs with serum (Leven et al. 1987). Addition of IL‐6 to the serum‐free cultures caused megakaryocytes to form extensive proplatelet extensions. We conclude that large numbers of pure guinea‐pig bone marrow megakaryocytes can be isolated by immunomagnetic bead selection, including low ploidy immature megakaryocytes. Spontaneous maturation occurred as evidenced by the increase in megakaryocyte size and ploidy. IL‐6 altered megakaryocyte size and morphology but not ploidy, indicating that these different characteristics of megakaryocytes may be regulated separately.
Difference in activity properties and subcellular distribution of neutrophil alkaline phosphatase between normal individuals and patients with trisomy 21
Authors: Grozdea J. , Vergnes H. , Brisson‐Lougarre A. , Bierme R. , Bourrouillou G. , Duchayne E. , Martin J. , Colombies P. 
Published: March 2008

SummaryBiochemical, cytochemical characteristics and electron microscopy subcellular distribution of neutrophil alkaline phosphatase (NAP) were analysed in blood and/or smear samples from 39 trisomy 21 patients (Down's syndrome) aged 11·5‐18 years (mean 15·5 years) and 55 normal subjects aged 12‐20·5 years (mean 17 years). All patients were karyotyped.NAP cytochemical procedures were carried out on all subjects: biochemical NAP determinations were made in 10 patients and 20 controls: ultrastructural electron microscopy of AP was performed in three patients and four normal subjects.Neutrophil alkaline phosphatase from patients with trisomy 21 displayed the following changes: (1) a significant increase of enzyme activity, (2) a high thermal lability of enzyme. Electron microscope morphology exhibited large deposits of NAP reaction product associated with the plasma membrane and intracellular main organelles, like phosphasomes. The NAP biochemical and cytochemical characteristics suggest that trisomy 21 neutrophils contain a nonspecific AP isoenzyme, closely related to the early placental form.
Variable X‐chromosome DNA methylation patterns detected with probe M27β in a series of lymphoid and myeloid malignancies
Authors: Hodges E. , Howell W. M. , Boyd Y. , Smith J. L. 
Published: March 2008

SummaryIn this study the X chromosome probe M27β was used to investigate DNA methylation at the DXS255 locus and hence X inactivation status and determination of tumour clonality in blood, bone marrow and biopsy tissue involved with morphologically and phenotypically defined lymphoid and myeloid disease from 14 female patients along with uninvolved bone marrow from two control individuals. Thirteen out of 16 individuals (81%) were restriction fragment length polymorphism (RFLP) heterozygous for DXSZ 55. DNA methylation status could not be assessed in the three DXS255 homozygous individuals. In eight DXS255 heterozygous individuals clonality was clearly demonstrated using M27β and in six of these cases independent analysis using T cell receptor (TcR) and immunoglobulin (Ig) gene probes confirmed the presence of clonal tumour cell populations. In the two controls, polyclonality was inferred from M27β probe analysis. In the remaining three cases (all acute lymphoblastic leukaemia (ALL)) both DXSZ55 X chromosome sequences appeared to be methylated. Clonality in these cases was demonstrated by TcR or Ig monoclonal gene rearrangements. These data demonstrate the value of the M27β probe for determining tumour clonality in a number of cases with lymphoid and myeloid disease but indicate that there may not always be a complete correlation between DNA methylation, X inactivation status and tumour clonality in certain lymphoid neoplasms, restricting the use of this probe in clonality studies. Correlations between DNA methylation, X inactivation status and stage of normal and neoplastic T and B cell development require further investigation.