bluuebunny/crossref_metadata_2025 huggingface dataset:
Plasma cell leukaemia of non‐producer type with missing light chain gene rearrangement
Authors: Tominaga Nobuhiko , Katagiri Shuichi , Hamaguchi Yasushi , Nishiura Tetsuo , Kanakura Yuzuru , Kanayama Yoshio , Nagao Kazuhiro , Kakiuchi Yoshiyuki , Nishida Kazuhiro , Abe Tatsuo , Honjo Tasuku , Yonezawa Takeshi , Tarui Seiichiro
Published: March 2008

Summary. A case of plasma cell leukaemia of non‐producer type is described. The patient presented with typical clinical features of plasma cell myeloma, including multiple osteolytic lesions, hypercalcaemia, renal failure and reduced polyclonal immunoglobulins, except that M‐component was not detected in either the serum or urine. Morphological examinations showed a plasmacytoid appearance of the neoplastic cells, while immunological studies failed to detect cytoplasmic immunoglobulin or secretory capacity. The surface phenotype of CD38+, PCA‐1+, DR‐, CD20‐, CD24‐, CD9‐, CD10‐ and surface immunoglobulin‐ was compatible with mature plasma cells. Chromosomal analysis showed the 14q+ marker due to translocation (6; 14) and deletion of the short arm of chromosome 1. Analysis of immunoglobulin genes revealed the presence of heavy chain gene rearrangement, but the light chain genes, both γ and λ remained in germline configuration. Such defective immunoglobulin gene rearrangement may be responsible for the failure of immunoglobulin biosynthesis and secretion by the neoplastic plasma cells. Furthermore, it is suggested that the morphological and phenotypic development of B cells may not necessarily depend on immunoglobulin light chain gene rearrangement, and that the oncogenic event in myeloma may occur at an earlier stage of B cell differentiation.
Bone marrow transplantation for chronic myeloid leukaemia in first chronic phase: importance of a graft‐versus‐leukaemia effect
Authors: Apperley J. F. , Mauro F. R. , Goldman J. M. , Gregory W. , Arthur C. K. , Hows J. , Arcese W. , Papa G. , Mandelli F. , Wardle D. , Gravett P. , Franklin I. M. , Bandini G. , Ricci P. , Tura S. , Iacone A. , Torlontano G. , Heit W. , Champlin R. , Gale R. P.
Published: March 2008

Summary.We analysed the incidence of graft failure, graft‐versus‐host disease (GVHD) and relapse of leukaemia in 208 patients undergoing allogeneic bone marrow transplantation (BMT) for chronic myeloid leukaemia in chronic phase in eight transplant centres in Europe and the United States. 106 patients received unmanipulated donor bone marrow (Group 1) and 102 patients received marrow depleted of T‐cells by incubation with the monoclonal antibodies Campath‐1 or CT‐2 and complement (Group 2). The incidence of graft failure was higher and of GVHD was lower in Group 2 than in Group 1. Relapse of leukaemia occurred more frequently in patients in Group 2 than in Group 1 (17v. 2, P<0.001). Multivariate analysis showed that the following factors were associated with an increased risk of relapse: the use of T‐cell depletion, the absence of GVHD and a high platelet count at the time of admission for transplant. The findings support the concept that a graft‐versus‐leukaemia effect mediated by T‐lymphocytes is important for cure of leukaemia after BMT.
Diagnostic and prognostic factors in acute monocytic leukaemia: an analysis of 51 cases
Authors: Scott Colin Stephen , Stark Allistair N. , Limbert Howard J. , Master Peter S. , Head Christine , Roberts Bryon E.
Published: March 2008

Summary. Diagnostic features (cytochemistry, immunophenotyping and serum biochemistry) were examined in 51 cases of acute monocytic leukaemia (AMoL). Peroxidase, Sudan black B and alpha naphthyl acetate esterase (ANAE) cytochemical reactions were unrelated to morphological (FAB groups M5a and M5b) or immunological subtype. ANAE cytochemistry, however, indicated that AMoL cases could be subdivided into those with typical (M‐type) reactions and those with insignificant staining or monocytic ANAE isoenzymes (defined by IEF). All cases were phenotypically CD13/CD33 positive and, with one exception, had > 30% HLA‐DR positive cells. Membrane CD14 expression was insignificant or variable in 33% of M5a cases in contrast to 23/24 M5b cases which showed high proportions of CD14‐staining cells with at least two monoclonal antibodies. Serum lysozyme, LDH and beta‐2 microglobulin (β2m) were increased in 88%, 68% and 81% of cases respectively but, with the exception of statistically higher lysozyme levels in CD14+ cases, were unrelated to the morphological, cytochemical or immunological diagnostic subgroups. Clinical and diagnostic features were also examined as possible prognostic indicators. The morphological, cytochemical and immunological subgroups of AMoL were not found to be of prognostic relevance but age (P=0.004), renal failure (P=0.005) and serum β2m levels (P=0.002) were related to patient survival. Moreover, renal failure and serum β2m remained significant (P=0.012 respectively) when age was taken into account and were shown to be independent prognostic variables.
Platelet surface charge heterogeneity: characterization of human platelet subpopulations separated by high voltage continuous flow electrophoresis
Authors: Crook M. , Crawford N.
Published: March 2008

Summary. Washed formol‐fixed normal human platelets have been separated into surface charge‐dependent subpopulations using high voltage continuous flow electrophoresis. The procedure is highly reproducible and the heterogeneity profile extends over 20–25 fraction tubes on the anodal side of the entry port to the separation chamber. Fractions have been subdivided into subpopulation pools A, B and C which have mean mobilities by analytical cytopherometry extending over the range 0.81–0.91 μm/s/V/cm from the least (C) to the most (A) electronegative cells. Coulter volume differences across the profile from 5.0 to 12.8 fl correlated well with electrophoretic mobilities whereas buoyant density appeared to be an independent parameter. Analysis of surface neuraminidase‐labile sialic acid of the platelets in pools A and C correlated well with differences in electrophoretic mobility, whereas a similar relationship for the alkaline phosphataselabile phosphate moieties (also believed to be contributory to cell surface electrokinetic properties) could not be established even though in both cases the profiles of the enzyme‐treated platelets showed significant shifts towards the cathode when compared with untreated cells. Titration of surface DTNB‐reactive sulphydryl (‐SH) groups revealed an inverse relationship between electronegativity and membrane ‐SH group status. This electrophoretic expression of subpopulation heterogeneity within the circulating platelet pool may have advantages in studying clinical conditions where the profiles may reflect cell surface interactions ‘in vivo’.
A human macrophage‐associated antigen (CD68) detected by six different monoclonal antibodies
Authors: Micklem Kingsley , Rigney Elizabeth , Cordell Jacqueline , Simmons David , Stross Paul , Turley Helen , Seed Brian , Mason David
Published: March 2008

Summary. Antibodies grouped together by the Third Workshop on Leucocyte Differentiation Antigens on the basis of pan‐macrophage reactivity on tissue sections were analysed in immunoprecipitation experiments. Antibodies Y2/131. EBM11, Ki‐M6 and Ki‐M7 all precipitated antigens of Mr 110000 which were shown to be identical by preclearing experiments. In addition a recently produced antibody, KP1, which identifies macrophages in paraffin‐embedded tissue, was shown to recognize the same antigen. The antibodies were tested on murine cells transfected with two clones, which had been isolated by screening a cDNA library with antibodies Y1/82A and EBM11. Cells transfected with the longer cDNA clone, coding for a molecule of Mr 110000, reacted with antibodies Y2/131. EBM11. Y1/82A and Ki‐M6, whilst the shorter clone, encoding a molecule of Mr 70000 gave the same result except that it did not induce expression of the Ki‐M6 epitope. KP1 antibody did not recognize any transfectants, possibly because of differences in glycosylation by the transfected cell line compared with human tissue. Five of the six antibodies appear to recognize different epitopes (the sixth, Ki‐M7, not having been evaluated in this way). It was concluded that these six antibodies react with a macrophage‐associated antigen for which the gene has been cloned. This group of antibodies has recently been designated CD68 by the Fourth Workshop on Human Leucocyte Differentiation Antigens.
Acute febrile neutrophilic dermatosis (Sweet's syndrome) in association with myelodysplastic syndromes: a report of three cases and a review of the literature
Authors: Soppi E. , Nousiainen T. , Seppa A. , Lahtinen R.
Published: March 2008

Summary. Three patients with acute febrile neurophilic dermatosis (AFND, Sweet's syndrome) in association with myelodysplastic syndromes (MDS) are described and, in addition, another 10 published cases are reviewed. In the reviewed patients the male/female ratio is 2:1. The great majority of the patients had decreased or normal leucocyte counts at the time the syndrome developed. Chromosome analysis was available in eight cases; in three the karyotype was normal while five patients had a clonal abnormality. No clinical or laboratory parameters seemed to predict the response to steroids; all but one of the treated patients responded promptly. Moreover, three patients recovered spontaneously. Shortly after the appearance of Sweet's syndrome nine patients developed acute leukaemia and one patient died with hypoplasia. Development of Sweet's syndrome in association with MDS may not be an uncommon clinical finding. The occasional MDS patient developing AFND will often progress into acute leukaemia and thus the occurrence of AFND warrants a bone marrow examination and close follow‐up of the patient.
Characterization of neoplastic and reactive cells in T‐cell lymphomas with cytogenetic, surface marker, and DNA methods
Authors: Lindholm Carita , Franssila Kaarle O. , Teerenhovi Lasse , Elonen Erkki , Peltomäki Päivi , Rapola Juhani , Ruutu Tapani , Saarinen Ulla , Knuutila Sakari
Published: March 2008

Summary. Three patients with T‐cell lymphoma were studied with a combined cytogenetic‐immunological analysis (MAC) as well as by Southern blotting analysis. The MAC method, which allows simultaneous study of both chromosomes and immunophenotype of a mitotic cell, revealed that in each case the karyotypically abnormal and, therefore, neoplastic cells originated from different stages of T‐cell maturation. In one case the abnormality was seen in cells expressing CD3 and CD10, in the second in cells expressing CD3 and CD8, and in the third case in cells expressing CD4 but not CD3. Mitoses were frequent also in non‐neoplastic cells, but these mitoses were normal. Southern blotting analysis revealed TCR beta rearrangements in all patients. In addition, one patient displayed TCR gamma rearrangement, and one patient a rearrangement in the IgH joining region.
Chimaerism following allogeneic bone marrow transplantation: detection of residual host cells using the polymerase chain reaction
Authors: Lawler M. , McCann S. R. , Conneally E. , Humphries P.
Published: March 2008

Chimaerism was assessed in five recipients following sex mismatched allogeneic bone marrow transplantation. Techniques included karyotyping of bone marrow cells, dot blot DNA analysis of blood and bone marrow suspensions, and in vitro amplification of DNA by the polymerase chain reaction (PCR) using blood and bone marrow suspensions and stored bone marrow slides. Results of karyotypic analysis suggested complete chimaerism in four patients, while in one patient mixed chimaerism was detected. Mixed chimaerism was also detected, however, in a second patient using PCR and confirmed by dot blot analysis on all tissues examined. PCR is a sensitive tool for investigation of chimaerism following bone marrow transplantation. Since this technique does not require radioactivity, it is an attractive method for use in a clinical laboratory. This technique represents a further development in the use of DNA methodologies in the assessment of haematological disease.
The effect of total body irradiation dose and chronic graft‐versus‐host disease on leukaemic relapse after allogeneic bone marrow transplantation
Authors: Frassoni Francesco , Scarpati Daniele , Bacigalupo Andrea , Vitale Vito , Corvo Renzo , Miceli Salvatore , Gualandi Francesca , Occhini Domenico , Clavio Marino , Van Lint Maria Teresa , Marmont Alberto
Published: March 2008

One‐hundred and five patients undergoing allogeneic bone marrow transplantation (BMT) for acute myeloid leukaemia (AML) (n = 61) and chronic myeloid leukaemia (n= 44) were analysed for risk factors associated with relapse. All patients received marrow from an HLA identical sibling after preparation with cyclophosphamide 120 mg/kg and total body irradiation (TBI) 330 cGy on each of the three days prior to transplantation. There was a difference of ±18% between the nominal total dose of 990 cGy and the actual dose received as indicated by dosimetric recordings. While interstitial pneumonitis had minimal impact on survival (4%) there was a considerable difference in the incidence of relapses. The incidence of relapse was 55% versus 11% in patients receiving less or more than 990 cGy respectively and this had a major impact on survival (38% v. 74% at 7 years) since transplant‐related mortality was comparable in the two groups. A multivariate Cox analysis indicated that a lower TBI dose (less than 990 cGy) was the most significant factor associated with relapse and the second most important factor associated with recurrence of leukaemia was the absence of chronic graft‐versus‐host‐disease (cGvHD). Actuarial relapse incidence was 62%, 28% and 18% for patients with no, limited or extensive chronic GvHD respectively. However, chronic GvHD had no significant impact on survival. Combined stratification for TBI dose and cGvHD showed that the dose effect of TBI on relapse was evident both in patients with and without cGvHD. Chronic GvHD influenced the risk of relapse only in patients receiving less than 990 cGy. These results suggest that a higher dose of TBI, within this schedule, produced long‐term disease‐free survival in the majority of AMLs and CMLs. Minor radiobiological side effects were experienced, but a small reduction of the dose may significantly increase the risk of relapse.
Bleeding time, blood groups and von Willebrand factor
Authors: Caekebeke‐Peerlinck Kathelijne M. J. , Koster Ted , Briët Ernest
Published: March 2008

The bleeding time in healthy volunteers was determined according to both the Ivy and the Simplate II techniques. A significantly longer bleeding time in people with blood group O than in people with non‐O blood groups was demonstrated with both techniques. This difference could not be attributed to a difference in sex ratio, platelet count or haematocrit. The mean level of von Willebrand factor in blood group O is lower than in non‐O blood groups, but we found no association between the level of von Willebrand factor and the bleeding time, despite the very broad range of von Willebrand factor levels in the subjects examined.
The natural history of human immunodeficiency virus infection in a haemophilic cohort
Authors: Lee Christine A. , Phillips Andrew , Elford Jonathan , Miller Elizabeth J. , Bofill Margarita , Griffiths Paul D. , Kernoff Peter B. A.
Published: March 2008

112 haemophilic patients infected with HIV were followed up with clinical and laboratory assessment between 1 December 1979 and 30 November 1988. Sixty‐six (59%) of the patients developed HIV‐related clinical symptoms and 22(20%) developed AIDS. Twenty (18%) of the patients developed p24 antigenaemia. Amongst the 59 patients whose date of seroconversion could be estimated the calculated 8‐year cumulative incidence of AIDS was 40% (symptoms 73%). For the whole cohort of 112 patients, the median slope of linear regression of the absolute T4 lymphocyte count was steeper for those with AIDS (‐0.113 × 109/1 per year) than for those without AIDS (‐0.054 × 109/1 per year) (P < 0.02). While 15 cases of AIDS developed during 58 patient‐years of follow up after falling below a T4 lymphocyte count of 0.2 × 109/1, only two cases occurred during 450 patient‐years before reaching this count. Thus the decline of the T4 lymphocyte count to 0.2 × 109/1 may be an appropriate additional end‐point for the assessment of new treatments for asymptomatic patients infected with HIV.
Heparin‐induced thrombocytopenia: mechanism of interaction of the heparin‐dependent antibody with platelets
Authors: Chong Beng H. , Fawaz Ismail , Chesterman Colin N. , Berndt Michael C.
Published: March 2008

The interaction of the heparin‐dependent antibody with heparin and platelets has been studied using the sera and purified IgG of four patients with heparin‐induced thrombocytopenia. Both normal platelets and Bernard‐Soulier syndrome (BSS) platelets which lack glycoprotein (GP) Ib, GPV and GPIX, aggregated in response to patient serum or IgG, but only in the presence of heparin. A monoclonal antibody (Mab) against platelet Fc II receptor (IV. 3) strongly inhibited the heparin‐dependent aggregation of both normal and BSS platelets induced by patient sera/IgG. Inhibition by the anti‐GPIb Mab (AK2) was variable and occurred only with normal platelets. Anti‐GPIX Mab (FMC 25) was not inhibitory with either normal or BSS platelets. Similar results were obtained using 14C‐serotonin release instead of platelet aggregation as a measure of platelet activation. These findings suggest that (1) the reaction of the heparin‐dependent antibody with platelets and heparin is mediated by a Fc‐dependent mechanism, (2) GPIb, GPV and GPIX are not involved in this reaction, and (3) the inhibitory effect of anti‐GPIb Mab on normal platelets is due to steric interference consistent with the platelet Fc receptor being in close proximity to GPIb.
CD 7 positive acute myeloid leukaemia: a subtype associated with cell immaturity
Authors: Coco Francesco Lo , Rossi Giulio De , Pasqualetti Daniela , Lopez Manuela , Diverio Daniela , Latagliata Roberto , Fenu Susanna , Mandelli Franco
Published: March 2008

Seventeen patients with acute myeloid leukaemia (AML) whose blasts co‐expressed the T‐cell associated CD7 antibody were identified among 160 consecutive AML cases. Fourteen had FAB defined AML according to morphocyto‐chemical criteria, whereas three patients were classified as ‘MO’ on the basis of immunophenotype. The incidence of CD7 positivity was particularly significant in the less differentiated subtypes MO and M1 compared with other FAB groups (P < 0.001). In all cases the myeloid determinants CD13 and/or CD33 were associated with CD7 expression. Other B‐lymphoid (CD10, CD19) or T‐lymphoid (CD2, surface and cytoplasmic CD3) markers were analysed and found to be negative. Five out of 15 cases examined were TdT+. Clonal rearrangements of the immunoglobulin heavy chain (IgH) and/or T‐cell receptor (TcR) β chain genes were identified in only three out of 13 cases. Among these, one out of five coexpressing TdT showed IgH rearrangement when analysed at the DNA level.

Clinical features at presentation and response to induction therapy did not allow us to consider CD7 + AML patients as a distinct subgroup with prognostic significance. Our data indicate that CD7 expression is a common finding in immature AML, being generally found in the absence of other T‐cell features. Rather than suggesting the occurrence of ‘mixed leukaemia’, such cases confirm a broader spectrum of CD7 reactivity and its possible identification of a particular subset of myeloid progenitors.
Multiple Xbal polymorphisms for carrier detection and prenatal diagnosis of haemophilia A
Authors: Chan Vivian , Tong T. M. F. , Chan T. P. T. , Tang Mary , Wan C. W. , Chan F. Y. , Chu Y. C. , Chan T. K.
Published: March 2008

Three Xbal restriction fragment length polymorphisms (RFLPs) can be detected using the factor VIII‐intron 22 probe (p482.6) in a Xbal‐KpnI double digest of genomic DNA. The Xbal (A) site had been reported by Wion et al (1986) to be in intron 22. while the two additional sites. Xbal (B) and Xbal (C), are shown here to be X‐linked and close to the Xbal (A) site. The frequencies of heterozygosity for these three sites are 0.49. 0.18 and 0.30 respectively. In 75 females the observed heterozygosity rate for the Xbal (A) site is 0.41 and this increased to 0.57 with the two additional sites. Care should be exercised when interpreting the Xbal RFLPs, since the 1.4 kb Xbal/Kpnl fragment and the 4.8 kb Xbal fragment are associated with both positive Xbal (A) and Xbal (B) sites. By the combined use of the multiple Xbal polymorphisms with the Bell site in intron 18, the carrier detection rate would increase to 67%. Four prenatal diagnoses had been performed using the multiple Xbal polymorphisms.
Bepridil protects sickle cells against the adverse rheological effects of cyclical deoxygenation
Authors: Johnston M. N. , Ellory J. C. , Stuart J.
Published: March 2008

Calcium influx into sickle cells, with consequential activation of the Ca2+‐activated K+ efflux (Gardos) channel, is a potential cause of cellular dehydration and loss of deformability. Bepridil, a recently described inhibitor of the Gardos channel, was found at pharmacological concentration (1 μmol/1) to inhibit significantly (P<0.01) the loss of deformability when sickle cells were subjected to cycles of oxygenation‐deoxygenation for 15 h at 37°C. Bepridil also inhibited significantly (P < 0.005) the formation of irreversibly sickled cells. Drugs that preserve the K+ and therefore water content of erythrocytes are of potential value for hydrotherapy of sickle cell disease.
Decreased amount of the Rh antigen D in hereditary spherocytosis (HS)
Authors: Szymanski I. O. , Araszkiewicz P. , Odgren P. , Snyder L. M.
Published: March 2008

This study was done to determine whether hereditary spherocytosis (HS) red blood cells (RBC) have decreased amounts of Rh antigens. Initially we studied the RBC of five members of one family, two of whom had HS. Using automated quantitative haemagglutination tests, we demonstrated that HS RBC agglutinated less with Rh antisera of four specificities than did normal RBC, indicating that Rh antigens are decreased on HS RBC. In this family, the strength of other blood group antigens on HS RBC was estimated by manual titres and agglutination scores. No appreciable differences in the agglutination of HS and normal RBC were observed with non‐Rh antisera. To assess the strength of the D antigen more accurately, the number of D sites was quantitated on the RBC of 19 individuals with HS and 11 of their healthy relatives. HS RBC had 9209±4084 (mean±SD) D sites, whereas the normal RBC had 15 394±5763 D sites. These two means were significantly different (P < 0.01). HS RBC were also compared to normal RBC of unrelated individuals who had the same Rh pheno‐type. These analyses showed that HS RBC had about half of the normal number of D sites. Our data indicate that HS red cells have decreased amount of the Rh antigen D and probably also of other Rh antigens.
Pregnancy associated aplastic anaemia: a report of five cases and review of current management
Authors: Aitchison R. G. M. , Marsh J. C. W. , Hows J. M. , Russell N. H. , Gordon‐Smith E. C.
Published: March 2008

The occurrence of aplastic anaemia in pregnancy has been long recognized but its rarity has made it difficult to establish the relationship between the two conditions and the optimal management. We now report five cases of aplastic anaemia in pregnancy and offer some recommendations for treatment. In two patients the pregnancy was allowed to continue to term and the disease persisted post‐partum leading to death in one case. The other three patients had their pregnancies terminated; one subsequently deteriorated and died, two had spontaneous remissions of their aplasia. We suggest that patients presenting with severe aplastic anaemia in early pregnancy should be offered termination because this may be followed by haematological improvement. If haematological improvement does not occur allogeneic bone marrow transplantation (BMT) may be considered. Aplastic anaemia presenting in late pregnancy should be treated with supportive care until delivery. On the basis of our experience, antilymphocyte globulin may safely be given during pregnancy.
Red blood cell aggregation and sedimentation: the role of the cell shape
Authors: Reinhart Walter H. , Singh Ajit , Straub P. Werner
Published: March 2008

The influence of erythrocyte shape changes on the sedimentation rate was studied in vitro and in vivo. In vitro the highest sedimentation rate was observed with a slight degree of stomatocytosis (morphological index – 0.3; i.e. one red cell out of three being a stomatocyte I). With increasing degrees of stomatocytosis the sedimentation rate gradually decreased. Echinocytosis reduced the aggregation and sedimentation very drastically; the sedimentation rate was virtually zero when echinocytosis I or higher degrees were present. The influence of abnormal cell shapes occurring in vivo was studied in patients with an abnormal blood smear. It was found that a severely abnormal red cell morphology reduced the sedimentation rate in a standardized, fibrinogen‐rich plasma to about half. These results indicate that the shape plays a crucial role in the aggregation and sedimentation of red cells and they may contribute to the understanding of the interaction of red cells with other cells such as endothelium.
The frequency of long‐term remission in patients with acute myelogenous leukaemia treated with conventional maintenance chemotherapy: a study of 760 patients with a minimal follow‐up time of 6 years
Authors: Preisler H. D. , Anderson K. , Rai K. , Cuttner J. , Yates J. , DuPre E. , Holland J. F.
Published: March 2008

Summary Remission duration associated with the administration of conventional maintenance chemotherapy to patients with acute myelogenous leukaemia was evaluated. The records of 760 patients who entered remission between 1974 and 1979 were reviewed. The median duration of remission was 1.1 years with 16% of patients remaining in remission at 8 years. The relapse curve was biphasic with a high rate of relapse during the first 2½ years of remission followed by a much lower relapse rate thereafter. Leukaemic relapses were noted through 8 years of remission. A plateau phase indicating freedom from the risk of leukaemic recurrence is not clearly apparent yet but may exist after the eighth year of remission.
Increase in the suppressor‐inducer T cell subset in multiple myeloma and monoclonal gammopathy of undetermined significance
Authors: Shapira Reuma , Froom Paul , Kinarthy Amalia , Aghai Esther , Lahat Nitza
Published: March 2008

Summary The expression of CD4 (helper‐inducer). CD8 (suppressor‐cytotoxic) and CD4 subpopulations (2H4 and 4B4) were studied in patients with multiple myeloma, monoclonal gammopathy of undetermined significance (MGUS) and in healthy controls. The percentages of CD4 + cells and CD8 + cells among total T cells were not different between the three groups studied. However the percentage of CD4 + cells of the suppressor‐inducer type (CD4 + 2H4 +) was 53 ± 9% in patients with MGUS, and 51 ± 9% in those with MM. comrpared to 46 ± 5 in the controls (P=0.033 and P= 0.07 respectively). A significant negative correlation between serum polyclonal IgM and the percentage of CD4 + 2H4 + cells was found in patients with MM but not in those with MGUS. No difference was found in the percentage of CD4+4B4+ (helper CD4+ cells) between the various groups. These findings suggest that the elevation of the suppressor‐inducer subset occurs prior to clinical manifestations of MM, perhaps as an immune response to the malignant clone. The existence of elevated proportions of CD4 suppressor‐inducer cells was associated with the hypogammaglobulinaemia observed in patients with MM. Since no hypogammaglobulinaemia was present in those with MGUS, additional factors are needed to explain the influence of the CD4 + 2H4 + cells on the production of immunoglobulins.
The application of X‐chromosome gene probes to the diagnosis of myeloproliferative disease
Authors: Lucas G. S. , Padua R. A. , Masters G. S. , Oscier D. G. , Jacobs A.
Published: March 2008

Summary X‐chromosome DNA probes for the phosphoglycerate kinase (PGK) and hypoxanthine phosphoribosyl transferase (HPRT) genes were used to study clonality in haemopoietic cells from 63 women with myeloproliferative disease, idiopathic erythrocytosis. secondary erythrocytosis or normal red cell volumes. A total of 25 women (39%) were heterozygous for one of the polymorphisms associated with these genes. Clonality was demonstrated in five out of six patients with polycythaemia vera (PV) and in three other patients with myeloproliferative disease. In all cases of PV, including the patient in whom clonality was not demonstrated, cultures of peripheral blood showed growth of endogenous erythroid colonies.
A plasma inhibitor of platelet aggregation in patients with Lassa fever
Authors: Cummins David , Fisher‐Hoch Susan P. , Walshe Karen J. , Mackie Ian J. , McCormick Joseph B. , Bennett Diane , Perez Gilda , Farrar Bertha , Machin Samuel J.
Published: March 2008

Summary Previous studies have shown that haemorrhage in Lassa fever is associated with abnormal in vitro platelet aggregation and a high mortality. In Sierra Leone we studied platelet aggregation in healthy local subjects, patients with laboratory‐confirmed Lassa fever and febrile patients in whom Lassa virus infection was excluded. There were no significant differences in the mean platelet counts of these groups. Patients with fulminant Lassa virus infection showed a gross depression of in‐vitro platelet responsiveness to 1 and 5 μm ADP and 4 μg/ml collagen compared to other groups (P=0·0004–0·0008 when compared to healthy controls, P=0·002–0·0008 when compared to mild Lassa fever patients). When plasma samples from five of these patients were mixed 1:1 with control platelet‐rich plasma, a marked inhibition of ADP‐induced aggregation was observed. No inhibitory activity was detected in plasma obtained from healthy subjects or febrile control patients. The presence of inhibitor was strongly associated with the occurrence of haemorrhage (P=0·03), depression of platelet aggregation (P=0·004) and severity of Lassa fever (P=0·007).
Antithrombin Sheffield: amino acid substitution at the reactive site (Arg393 to His) causing thrombosis
Authors: Lane D. A. , Erdjument H. , Flynn A. , Di Marzo V. , Panico M. , Morris H. R. , Greaves M. , Dolan G. , Preston F. E.
Published: March 2008

A Sheffield family with a predisposition towards thrombosis has been shown to have a functional abnormality of antithrombin. The abnormality was detected as reduced heparin cofactor activity, with normal antigenic levels of antithrombin. Crossed immunoelectrophoresis performed in the absence and presence of heparin was normal. The antithrombin was isolated by heparin Sepharose affinity chromatography. It had normal mobility on SDS polyacrylamide gel electrophoresis. However, the second order rate constant of inhibition of thrombin was about half that of normal, and this was compatible with a heterozygous abnormality involving the reactive site. The antithrombin was further purified by chromatography on thrombin‐Sepharose (to remove the normal component), reduced, S‐carboxymethylated and fragmented with cyanogen bromide. A pool containing the reactive site region was digested with trypsin and the molecular size of peptides generated determined by fast atom bombardment mass spectrometry. The two peptides adjacent to the Arg393‐Ser394 bond of mass 2290 and 700 were almost absent from the mass spectrum, but an additional peptide of mass 2952 was present. Subdigestion with V8 protease reduced the mass of this peptide to 1748. These peptides generated by trypsin and V8 protease were almost identical to those obtained when another variant, antithrombin Glasgow, was treated in the same way (Erdjument et al, 1988). It is concluded that the molecular abnormality of antithrombin Sheffield is identical to that of antithrombin Glasgow, Arg393 to His.
Human monoclonal anti‐D antibodies. I. THEIR PRODUCTION, SEROLOGY, QUANTITATION AND POTENTIAL USE AS BLOOD GROUPING REAGENTS
Authors: Kumpel B. M. , Poole G. D. , Bradley B. A.
Published: March 2008

Eight monoclonal IgG antibodies to the Rh antigen D, produced by Epstein‐Barr virus transformed B‐lymphoblastoid cell lines from two individuals, have been assessed for their suitability as blood grouping reagents. All showed similar specificity and agglutinated all red cells with partial D antigens tested except category DVI cells. They gave strong reactions with Du red cells in indirect antiglobulin tests, and they all gave good reactions with R1R1, R2R2 and R0r cells in manual tests using antiglobulin, enzyme or albumin methods. Initial studies showed that some of the monoclonal antibodies worked well at high dilutions on a Technicon Autogrouper 16C when used for routine D‐typing of blood donors. Antibody production by the cell lines was stable for many months in continuous culture. These monoclonal antibodies may be useful diagnostic reagents.
Pituitary dependence of bone marrow function
Authors: Nagy E. , Berczi I.
Published: March 2008

Summary. The anaemia, leucopenia, thrombocytopenia and impaired DNA and RNA synthesis in the bone marrow of hypophysectomized rats could be restored by syngeneic pituitary grafts placed under the kidney capsule, or by treatments with ovine or bovine prolactin or growth hormone. Treatment with ACTH, FSH, LH and TSH had no effect in this respect. These results indicate that bone marrow function is regulated by the pituitary gland.
Circulating monoclonal B lymphocytes in multiple myeloma
Authors: Chiu E. K. W. , Ganeshaguru K. , Hoffbrand A. V. , Mehta A. B.
Published: March 2008

Summary. Peripheral blood mononuclear cells from 28 patients with multiple myeloma (MM) and nine patients with monoclonal gammopathy of unknown significance (MGUS) were studied by immunoglobulin gene analysis. Clonal immunoglobulin gene rearrangements in peripheral blood mononuclear cells (PBIGRA) were demonstrated in 10 of the 28 MM patients (36%). Bone marrow and peripheral blood mononuclear cells were studied simultaneously in five of these 10 patients, and identical gene rearrangements were demonstrated in both. The incidence of such gene arrangements was higher in patients with active disease (cases at presentation or relapsed = 10/19 [47%]) compared to remission status (0/9) and higher in untreated (47%) compared to treated patients (11%) (P<0.05). Patients with this phenomenon had higher serum calcium levels (P<0.001), and higher bone marrow plasma cell counts (P<0.05). Serum creatinine and β2‐microglobulin were also higher but did not reach statistical significance. None of the patients with monoclonal gammopathy of uncertain significance had gene arrangements. Our findings confirm that circulating B lymphocytes are part of the malignant clone in MM and their presence correlates with high tumour volume.
The effect of fresh versus stored blood on post‐operative bleeding after coronary bypass surgery: a prospective randomized study
Authors: Wasser M. N. J. M. , Houbiers J. G. A. , D'Amaro J. , Hermans J. , Huysmans H. A. , Kinijnenburg G. C. van , Brand A.
Published: March 2008

Summary. In a prospective study, 237 patients undergoing a primary coronary bypass operation were randomized to receive 2 units of fresh whole blood (study group) or stored (2–5 d) blood (control group) at the end of the extracorporeal circulation.

Serious post‐operative bleeding necessitating a re‐thoractomy occurred in 4.2% of all patients with an equal distribution over the two groups. Post‐operative haemoglobin content and platelet counts were higher in the study group, but the differences were small and clinically not important. There were no differences in transfusion requirements, post‐operative blood losses and haemostatic parameters between the trial groups.

At low post‐operative platelet counts (below 120.109 platelets/1) however, patients in the control group lost significantly more blood and had increased transfusion requirements compared with patients in the study group (7.1 versus 4.8 units). These differences must be attributed to qualitative platelet defects in the transfused units of stored blood.

The small, clinically insignificant, differences in two laboratory parameters between the study and control groups, and the increased transfusion requirements of a subpopulation of patients with low platelet counts in the control group do not justify giving fresh blood or prophylactic platelet transfusions to coronary bypass patients.
Circadian rhythm of erythropoietin in human serum
Authors: Wide L. , And C. Bengtsson , Birgegárrd G.
Published: March 2008

Summary. When a sensitive radioimmunoassay for erythropoietin in serum (S‐Epo), using labelled recombinant hEpo and antiserum to urinary hEpo, was evaluated for clinical use, a large intra‐individual variation was observed during a 24 h period. The possibility of a circadian rhythm with a variation during day time which has to be considered for clinical evaluation of the S‐Epo level was investigated in 27 hospitalized adult patients. They had various illnesses, but all had a level of S‐Epo at 08.00 or 12.00 hours within the reference range of healthy individuals and normal cortisol levels during the 24 h. A well‐marked circadian rhythm of S‐Epo was found with the lowest mean level at 08.00 hours, a 42% increase at 16.00 hours and a 60% increase to the highest at 20.00 hours. Twelve (44%) of the 27 patients, with S‐Epo levels within the reference range (3.3–13.5 IU/l) at 08.00 or 12.00 hours, had levels above this range at 16.00 hours. For clinical evaluation of the S‐Epo level, the time of day for collection of the blood specimen has to be taken into consideration. It is recommended that blood specimens are taken in the morning between 07.30 and 12.00 hours. The mechanisms behind the observed circadian rhythm of S‐Epo are unknown.
Immunochemical characterization of the new platelet alloantigen system Bra/Brb
Authors: Santoso Sentot , Kiefel Volker , Mueller‐Eckhardt Christian
Published: March 2008

Summary. We report the immunochemical characterization of the new platelet‐specific alloantigens Bra and Brb. Bra antibodies were from mothers of children with neonatal alloimmune thrombocytopenia (NAIT), and anti‐Brb was found in the serum of a polytransfused patient. By radioimmunoprecipitation, anti‐Bra and anti‐Brb precipitated two proteins with apparent relative molecular masses in sodium dodecyl sulphate‐polyacrylamide gel electrophoresis of 155000 and 130000 under non‐reduced conditions, and of 165000 and 148000 under reduced conditions. In two‐ dimensional polyacrylamide gel electrophoresis, the two bands moved with isoelectric points ranging from 5·2 to 5·4 and from 4·5 to 4·7, respectively. These features fulfil previously defined criteria for platelet membrane glycoproteins (GP) Ia and IIa. The results were supported by data obtained by an assay employing monoclonal antibody (mab)‐specific immobilization of platelet antigens (MAIPA). By this technique, Bra and Brb antigens could be immobilized by mabs specific for the GP Ia/IIa complex (mab Gi 14) or a mab specific for the very late antigen‐2 (mab 12 F1), but not by a mab specific for GP IIb/IIIa complex (mab Gi3). Furthermore, platelets from a thrombasthenic patient with complete absence of GP IIb/Ea expressed the Brb antigen normally as shown in MAIPA; this antigen could be immunoprecipitated with anti‐Brb and was identical to that of normal platelets. This confirms that the antigens of the Br system are not associated with the GP IIb/IIIa complex. By direct binding studies using mabs Gi3 and Gi14, we calculated that 51500±3900 molecules of anti‐GP IIb/IIIa and 6470±500 molecules of anti‐GP Ia/IIa were bound per platelet at saturation. Our results provide evidence for the first platelet‐specific alloantigen system residing on the GP Ia. The difficulty in detecting anti‐Bra and anti‐Brb by direct binding assays may be related to the small number of GP Ia/IIa complexes on platelets.
The reappraisal of dilute tissue thromboplastin inhibition test in the diagnosis of lupus anticoagulant
Authors: Liu H. W. , Wong K. L. , Lin C. K. , Wong W. S. , Tse P. W. T. , Chan G. T. C.
Published: March 2008

Summary. The dilute tissue thromboplastin inhibition (DTTI) test (Schleider et al. 1976) is a sensitive but non‐specific test for lupus anticoagulant (LA). False positive results are seen in patients with clotting factor deficiency involving the extrinsic pathway and also in some patients with specific factor inhibitors (Triplett et al. 1983; Rosove et al. 1986). Since the effect of LA is phospholipid dependent but those of factor deficiency and specific inhibitors are not, we analyse the test results by comparing the degree of inhibition using different dilutions of tissue thromboplastin and express it as the DTTI index. This is defined as the clotting time ratio with 0·2% tissue thromboplastin divided by the clotting time ratio with 2% tissue thromboplastin. We also perform a dilute tissue thromboplastin time with platelet substitution to see if this could neutralize the inhibition caused by LA. Both of these modifications can reliably distinguish LA from other conditions associated with prolonged APTT better than the original DTTI test.
Alteration of the mechanical properties of sickle cells by repetitive deoxygenation: role of calcium and the effects of calcium blockers
Authors: Nash G. B. , Boghossian S. , Parmar J. , Dormandy J. A. , Bevan D.
Published: March 2008

Summary. The formation of dense, poorly deformable sickle cells was studied by subjecting pre‐separated, less dense cells to repeated deoxygenation and reoxygenation for 15 h. In the presence of Ca (2 mmol/l), this process caused the number of irreversible sickled cells to increase five‐fold, mean cell haemoglobin concentration to increase by 13% and cellular potassium to decrease by 22%. Also, red cell filterability through 5 μm filter pores was greatly worsened. These effects decreased but were not totally abolished when the extracellular Ca concentration was lowered to zero or 0·01 mmol/1. If a high K medium was used (135 mmol/l), cell swelling rather than shrinkage occurred. Swelling also occurred if ouabain was added to the incubation. The Ca‐channel blockers nitrendipine and nisoldipine had different effects. Nitrendipine. in the range 10−7‐10−5 mol/l, was partially protective against all the induced changes, but nisoldipine was not protective at 10−8 or 10−6 mol/l. Thus, deterioration in the properties of sickle cells appears to be linked to Ca‐dependent potassium loss during repeated sickling and is inhibited by nitrendipine.
Phenotypic changes on hairy cells exposed in vitro to interferons: a quantitative FACS study
Authors: Till K. J. , Cawley J. C.
Published: March 2008

Summary. In vitro culture of hairy cells (HC) (five patients) with αIFN (100 U/ml) significantly enhanced MHC and CD22 antigen expression and reduced CD25, sIg and FMC7 positivity, together with consistent but not significant reductions in CD9, 19 and HC2. A sixth patient, who was refractory to the effects of αIFN in vivo, was also studied and none of these changes were seen in her HC when cultured with αIFN in vitro. γIFN (100 U/ml) produced a much less marked increase in MHC and reduced sIg. TPA (100 ng/ml) induced changes very similar to those observed with αIFN, α and γIFN had no effect on a range of other antigens in HCL (including CD23 and 38, and transferrin receptors) and also produced no significant antigenic changes in CLL cells.

It is concluded that these findings are compatible with partial activation/maturation of HC by αIFN. It is suggested that the specificity of the phenotypic changes induced on HC by αIFN is a consequence of HC representing a stage of B‐cell development at which αIFN has specific immunomodulatory effects.
Human immunodeficiency virus detection: correlation with clinical progression in the Edinburgh haemophiliac cohort
Authors: Cuthbert R. J. G. , Ludlam C. A. , Rebus Selma , Peutherer J. F. , Aw D. W. J. , Beatson Dianne , Steel C. M. , Reynolds Billie
Published: March 2008

Summary. HIV p24 antigenaemia and virus detection in cultures of peripheral blood lymphocytes were examined in 16 of 18 haemophiliacs infected with HIV by a single batch of Scottish National Blood Transfusion Service factor VIII concentrate. Six (38%) had p24 antigenaemia and 11 (69%) had positive lymphocyte cultures. All seven patients with serious HIV disease (CDC group IV) had positive lymphocyte cultures whereas four (57%) had p24 antigenaemia. Four of nine (44%) patients with asymptomatic HIV disease (CDC groups II and III) had positive cultures and two (22%) had p24 antigenaemia. Twenty‐eight of 36 samples from the symptomatic group were HIV culture positive compared with nine of 30 samples from the asymptomatic group (P < 0·001). None of 14 antibody negative haemophiliacs who also received the implicated batch of factor VIII had p24 antigenaemia or positive HIV cultures.

The ability to detect HIV in cultured lymphocytes correlates with the clinical severity of HIV disease in this cohort.
Heterogeneity of fibrinogen receptor expression on platelets activated in normal plasma with ADP: analysis by flow cytometry
Authors: Jackson Carl W. , Jennings Lisa K.
Published: March 2008

Summary. Fibrinogen receptor expression of platelets activated in normal plasma by ADP was measured by flow cytometry after labelling bound fibrinogen with fluorescein‐conjugated antifibrinogen antibody. The platelet response to ADP was heterogeneous both with respect to number of platelets binding fibrinogen and the amount of fibrinogen bound per platelet. The proportion of platelets showing positive antifibrinogen antibody binding increased with increasing ADP concentration; however, even at 10−3m ADP, usually about one‐fifth of the platelets failed to demonstrate bound fibrinogen. The non‐responsive platelets tended to be the smaller ones. The relative fluorescence intensity of individual platelets also increased as ADP concentration was increased, indicating that the average number of fibrinogen molecules bound was also related to agonist concentration. The amount of fibrinogen bound following platelet activation directly correlated with the quantity of surface glycoprotein IIb detected by Tab antibody and with platelet size. This study demonstrates that platelet response to ADP in native plasma is heterogeneous in both the proportion of platelets activated and in the number of available fibrinogen receptors per platelet. This heterogeneity is related to platelet size and glycoprotein IIb‐IIIa content. These observations indicate that models of ligand interaction with membrane receptors on intact cells requiring an exposure step must take into account the heterogeneity of response within a cell population. In addition to providing new insights into the response of individual platelets to activation, these results suggest that study of platelet bound fibrinogen by flow cytometry may be useful for the detection of platelet activation in vivo.
Prevalence of monoclonal Ig with λ light chains in chronic myelocytic leukaemia
Authors: Guglielmi P. , Davi F. , Brouet J. C.
Published: March 2008

Light chains from seven of eight monoclonal Ig detected in the serum of patients with chronic myelocytic leukaemia (CML) were of the λ isotype. Although the presence of monoclonal Ig in CML is distinctly unusual, this prevalence of λ chains is unlikely to be a mere coincidence. All patients had otherwise typical clinical, chromosomal and molecular (bcr rearrangements) features of classical CML. The λ locus appeared unaltered. Only two patients had an excess of dystrophic plasma cells suggestive of multiple myeloma.