bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic thrombocytopenia with autosomal dominant transmission: a review of 54 cases
Authors: Najean Yves , Lecompte Thomas
Published: March 2008

On the basis of a retrospective study of 3600 platelet kinetic studies, we have isolated 54 cases with chronic thrombocytopenia, a normal autologous and homologous platelet lifespan, and increased mean platelet volume without Döhle bodies, the absence of any functional platelet abnormalities, and a normal megakaryocyte count. These cases were either discovered during the first few years of life (i.e. constitutional) and/or were proven to be familial (autosomal dominant transmission). Previous treatments (corti‐costeroids, immunoglobulins, androgens, immunosuppres‐sor agents, splenectomy) were not effective in any of these cases or in their relatives.

A new syndrome can therefore be proposed which can easily be suspected on the basis of platelet kinetic studies performed in cases of early onset, increased platelet volume, failure of corticosteroids or evidence of a familial blood disorder. It can be proved when the autologous platelet life span is demonstrated to be normal in spite of a chronic thrombocytopenia and a normal megakaryocytic count. The recognition of this syndrome will avoid neonatal complications (cephal‐haematomas), surgical complications, and the use of expensive and possibly harmful ineffective treatments, both in the propositus and in other abnormal family members.

The syndrome is certainly frequent (54 cases are presented here), but the diagnosis is often missed or delayed due to the low risk of haemorrhage. However, it is associated with a certain risk of leukaemia (four cases in three families).
Detection of immunoglobulin gene rearrangement in B lymphoid malignancies by polymerase chain reaction gene amplification
Authors: Deane Miriam , Norton John D.
Published: March 2008

Immunoglobulin heavy chain gene rearrangement serves as a marker of cell lineage and clonality in B lymphoproliferative disorders. We have used polymerase chain reaction (PCR) gene amplification to detect immunoglobulin gene rearrangements involving VH251. a heavy chain variable region preferentially utilized in B lymphoproliferative disorders. Using synthetic amplimers derived from VH251 and the heavy chain joining region, under conditions of high stringency, a homogeneous VH251‐specific fragment of approximately 350 bp could be amplified from leukaemic DNA. Of 53 cases of B lineage acute lymphoblastic leukaemia screened for VH2 51 rearrangement by PCR, 10 were positive. A background level of VH251 rearrangement could also be amplified from normal peripheral blood and bone marrow DNA. but a VH251 rearranged leukaemic clone representing 0‐01% of bone marrow mononuclear cells could be readily detected. The application of PCR to detect immunoglobulin gene rearrangement involving VH251 and potentially other preferentially utilized V regions provides a sensitive method both for tracking malignant B cells and for the study of normal B cell developmental pathways through which B lineage malignancies arise.
Elevated white blood cell synthesis of leukotriene C4 in chronic myelogenous leukaemia but not in polycythaemia vera
Authors: Stenke Leif , Samuelsson Jan , Palmblad Jan , Dabrowski Lech , Reizenstein Peter , Lindgren JanÅke
Published: March 2008

Leukotriene (LT) formation was studied in ionophore A23187‐stimulated white blood cell (WBC) preparations from patients with chronic myelogenous leukaemia (CML; n=14), polycythaemia vera (PV; n=10) and two control groups consisting of patients with non‐malignant inflammatory disease (n= 4) and normal healthy donors (n= 25). The synthesized products were identified and quantitated using high‐performance liquid chromatography combined with computerized UV‐spectroscopy.

White blood cell preparations from the CML patients produced more LTC4 (40.2±7.9 pmol/106 WBC, mean±SEM) than WBC from the healthy donors (9.0±1.8), P<0.0005. In contrast, the formation of LTB4 was normal and there was no increase in the total leukotriene synthesis (the sum of LTC4, LTB4, 20‐OH‐LTB4 and the Δ6‐trans‐isomers of LTB4). The ratio between leukotrienes C4 and B4 was strongly elevated in the CML group; 1.67 ± 0.25 v. 0.37±0.07 in the controls, P<0.0005. No significant correlation was observed between the levels of LTC4 and the number of known LTC4 producing cells (such as monocytes, eosinophils and basophils) in the CML WBC preparations. In contrast, a correlation was found between the sum of neutrophilic granulocytes and metamyelocytes in these suspensions and the amount of LTB4 formed; r=0.600. P<0.05 A number of other laboratory or clinical variables of the CML patients (including total white blood cell and platelet counts, differential counts, previous cytotoxic treatment, time from diagnosis, time from last treatment, post study survival and age) did not significantly correlate with the formation of leukotrienes. No abnormality in the production of LTB4 or LTC4 was observed in granulocyte and WBC preparations from the patients with polycythaemia vera and non‐malignant inflammatory disease, respectively. The results indicate a selectively increased LTC4 producing capacity in CML.
Use of prostaglandin E1 for prevention of liver veno‐occlusive disease in leukaemic patients treated by allogeneic bone marrow transplantation
Authors: Gluckman E. , Jolivet I. , Scrobohaci M. L. , Devergie A. , Traineau R. , Bourdeau‐Esperou H. , Lehn P. , Faure† P. , Drouet L.
Published: March 2008

Prostaglandin E1 (PGE1) was used to prevent veno‐occlusive disease (VOD) of the liver after allogeneic bone marrow transplantation (BMT) for leukaemia. It was given in continuous i.v. infusion from day – 8 to day 30 after BMT at a dose of 0.3 μg/kg/h. The patients were studied according to the risk factors for VOD: diagnosis, intensification of the conditioning and previous liver abnormalities. The diagnosis of VOD was made on at least two of the following factors: weight gain, hepatomegaly, jaundice, ascitis, pain of the right upper quadrant, increased platelet consumption. 109 patients were studied, 50 were treated by PGE1 and 59 did not receive it. The actuarial incidence of VOD was 12.2% in the PGE1 group and 25.5% in the non PGE1 group (P=0.05). In acute leukaemia, the incidence was 39.1% in the non‐treated group and 12.8% in the PGE1 treated group (P=0.02). Patients with previous hepatitis had an incidence of 62.5% in the non treated group and 15.5% in the treated group (P=0.05). A positive cytomegalovirus (CMV) serology seemed to increase the risk of VOD: the incidence of VOD was 31.4% in non‐treated patients and 22% in PGE1 treated patients. The multivariate analysis of the risk factors for VOD shows that unfavourable factors were: recipient positive CMV serology (P=0.06), hepatic disease prior to transplant (P=0.02) and the absence of PGE1 treatment (P=0.02). This study suggests that prophylactic PGE1 treatment may decrease the incidence of VOD in patients treated for leukaemia by allogeneic bone marrow transplantation.
The effect of molecular weight on heparin binding to platelets
Authors: Horne McDonald K. , Chad Elizabeth S.
Published: March 2008

Low molecular weight heparin is reported to be less reactive with platelets than larger heparins. We probed the molecular basis for this pattern of reactivity by characterizing the saturable platelet binding of [3H]heparin in plasma using heparins of different molecular weights (Mr˜ 3000, ˜ 5000, ˜ 10 000, ˜ 15000). Binding affinity increased with increasing molecular weight, as expressed by decreasing apparent dissociation constants (Kdapp˜ 1.3 μM for Mr˜ 3000, to Kdapp˜ 0.31 μM for Mr˜ 15000). After adjusting for the effect of antithrombin III in the plasma, true dissociation constants (Kd) could be calculated and these showed the same trend with molecular weight (Kd˜ 1.1 μM for Mr ˜ 3000 to Kd˜ 0.096 μM for Mr˜ 15 000). Platelet binding capacity for the different heparin fractions also increased with molecular weight, although this correlation appeared to lessen with the largest species. Heparin antithrombin III affinity was shown not to affect heparin binding to platelets. We propose a model in which heparin binding to platelets is mediated by charge interaction. Larger molecules with more charge bind with greater affinity and to sites with a broader range of electronegativity than do smaller, less
Platelet aggregating IgG antibody to platelet surface glycoproteins associated with thrombosis and thrombocytopenia
Authors: Pfueller Sharron L. , David Rosemary , Firkin Barry G. , Bilston Robyn A. , Cortizo W. Fernando , Raines Geoffrey
Published: March 2008

Previously described platelet‐aggregating antibodies associated with thrombosis and thrombocytopenia required heparin for their in vivo and in vitro expression. We have observed a patient with thrombosis who became thrombocytopenic during heparin treatment, but who suffered further thrombotic events and continued thrombocytopenia for 3 months after heparin withdrawal. The patient's plasma contained a potent platelet aggregating factor reactive with both his own and normal platelets in the absence of heparin. It also caused [14C]serotonin secretion from labelled platelets from normal donors and patients with either Glanzmann's thrombasthenia or Bernard‐Soulier syndrome. This factor was an IgG and was neutralized by antibody specific for IgG lambda light chains. While the patient was thrombocytopenic an IgG paraprotein with lambda light chains was detected by isoelectrofocussing. After corticosteroid treatment it disappeared and the patient recovered. The active, but not the recovery serum contained IgG which immunoprecipitated a glycoprotein with characteristics of Glycoprotein IV from platelets labelled with Na[3H]BH4/periodate. Thus platelet‐aggregating IgG antibodies with direct specificity for platelet surface glycoproteins may be associated with thrombosis/thrombocytopenia.
Myelofibrosis with myeloid metaplasia: clinical and haematological parameters predicting survival in a series of 133 patients
Authors: Visani G. , Finelli C. , Castelli U. , Petti M. C. , Ricci P. , Vianelli N. , Gianni L. , Zuffa E. , Aloe Spiriti M. A. , Latagliata R. , Pileri S. , Magrini U. , Gugliotta L. , Morra E. , Bernasconi C. , Mandelli F. , Tura S.
Published: March 2008

The prognostic value of 12 clinical and haematological parameters, recorded at diagnosis, in myelofibrosis with myeloid metaplasia (MMM) was retrospectively analysed in a consecutive series of 133 patients followed for a minimum of 60 months. Multivariate analysis showed that the following features were associated with a significantly shorter survival: (1) short period of time (<13 months) between first symptoms and diagnosis; (2) anaemia (haemoglobin <10 g/dl); (3) leucocyte count >12 × 109/l; (4) peripheral blood granulocyte precursors >10%. Age, splenectomy and percentage of peripheral blood metamyelocytes were found significantly to affect survival only from univariate analysis, whereas sex, size of spleen, thrombocytopenia and thrombocytosis were of no prognostic significance. These data suggest that a more intensive chemotherapy might be useful for younger patients with bad prognostic factors at diagnosis.
Acute monocytic leukaemia in adults: treatment and prognosis in 99 cases
Authors: Fenaux Pierre , Vanhaesbroucke Colette , Estienne Marie Hélégne , Preud'homme Claude , Pagniez Dominique , Facon Thierry , Millot Frédéric , Bauters Francis
Published: March 2008

Acute monocytic leukaemia (AMoL) was diagnosed in 99 adults, aged 18–85 years (median 56) over a period of 10 years. Sixty‐five patients had extramedullary leukaemia, 13 had clinical signs of leucostasis, and 19 had disseminated intravascular coagulation. Four patients died before receiving any treatment, 12 received supportive care only and seven received low dose AraC, but only one of them responded. Seventy‐six patients received intensive chemotherapy, 72 of them with an anthracycline‐AraC based regimen, with or without an epipodophyllotoxin.

Fifteen patients died within 7 d of diagnosis, due to leucostasis in nine cases. Predictive factors for early death were advanced age, leucostasis, fever, leucocytes above 100 x 109/l, and renal failure. Fifty (66%) of the patients treated intensively reached complete remission (CR). Advanced age, fever and complex cytogenetic abnormalities were significantly associated with a lower CR rate. Median actuarial disease‐free survival was 20.5 months, and was not significantly influenced by any pretreatment parameter. Five patients relapsed in the central nervous system (CNS), in spite of systematic CNS prophylaxis. No differences in CR rates were seen with the three anthracycline–AraC based regimens used in our patients. Significant differences in disease‐free survival were seen between them, however, suggesting that early consolidation chemotherapy and, more hypothetically, epipodophyllotoxin agents could prolong remission duration in AMoL.
Increased red blood cell aggregation in retinal vein occlusion
Authors: Chabanel Anne , Glacet‐Bernard Agnégs , Lelong Françloise , Taccoen Alain , Coscas Gabriel , Samama Meyer M.
Published: March 2008

Reversible aggregation of red blood cells (RBC) plays an important role in determining the flow properties of blood, and is the cause of the increase in blood viscosity at low shear rates. Retinal venous circulation is characterized by the combination of a low flow state and a high vascular resistance which might severely limit its capacity to adjust to high blood viscosity. These characteristics make the venous circulation in the retina particularly dependent on haemorheological factors. To test the possibility that high RBC aggregation could predispose to the onset and development of retinal vein occlusion (RVO), RBC aggregation and disaggregation (SEFAM erythroaggregameter, France) were measured in 64 patients with RVO. Results were compared to those of a group of 64 controls, similar in age, sex, smoking habit and associated pathologies. Increased RBC aggregation was observed in 52% of the patients, and the mean values showed a highly significant elevation of RBC aggregation parameters in RVO patients (+ 14%) when compared with controls (P<0.001). Subgroups were compared to study the influence of site (central versus branch), form (ischaemic versus non‐ischaemic), duration and severity of the occlusion on the aggregation parameters. No significant differences were found between these various subgroups. An increase in RBC aggregability and in the shear resistance of RBC aggregates, by predisposing to circulatory stasis, is likely to contribute to the onset of RVO.
Hb Icaria–Hb H disease: identification of the Hb Icaria mutation through analysis of amplified DNA
Authors: Efremov G. D. , Josifovska O. , Nikolov N. , Codrington J. F. , Oner C. , Gonzalez‐Redondo J. M. , Huisman T. H. J.
Published: March 2008

Hb Icaria–Hb H disease was observed in a Yugoslavian teenager who exhibited moderate anaemia with severe microcytosis and hypochromia and 16% Hb H. Four of his relatives were Hb Icaria heterozygotes; their haematological data were comparable to those with a deletional type of α‐thalassaemia‐2. The patient also had an additional α‐thalassaemia‐1 deletion, an ∼20.5 kb deletion, common among Mediterranean populations. The Hb Icaria mutation, i.e. the TAAÁA mutation at codon 142, was identified by hybridization of amplified DNA with specific probes. The mutation is located on the α2‐globin gene; the one remaining α1‐globin gene is apparently able to compensate sufficiently for the loss of the three α‐globin genes to maintain a haemoglobin level of 8–9 g/dl.
Murine monoclonal antibodies against a unique determinant of erythrocytes, related to Rh and U antigens: expression on normal and malignant erythrocyte precursors and Rhnull red cells
Authors: Kr. vondem Borne Albert E. G. , Bos Marjolein J. E. , Lomas Christine , Tippett Patricia , Bloy Christian , Hermand Patricia , Cartron Jean Pierre , Admiraal Lucia G. , Van de Graaf Jettie , Overbeeke Marijke A. M.
Published: March 2008

Three murine monoclonal antibodies (Mabs) MB‐2D10, LA‐18.18 and LA‐23.40 were prepared. They reacted with red cells of all common and most rare blood‐group phenotypes, with the exception of those of the RhnullU negative and RhmodU negative phenotypes. So far, only a single example of an alloantibody (Duclos or anti‐Rh38) of a similar specificity has been found.

Serological studies indicated that the Mabs were probably not directed against an antigenic determinant of Rh polypeptides, the LWab glycoprotein or glycophorin B, all structures absent from or aberrantly expressed on Rhnull red cells. The antigen was found to be erythrocyte‐specific, and was also present on pro‐erythroblasts, erythroblasts and malignant erythroblastoid cells but not on erythroid progenitors in the bone marrow.

The Mabs were found to block each other in an immune rosette method and are thus probably directed against the same epitope or against neighbouring epitopes on the same structure.

In immunochemical studies, MB‐2D10 precipitated the 30–32 kDa Rh polypeptides from red cell membranes and a protein or proteins which formed diffuse and overlapping bands in SDS–polyacrylamide gel electrophoresis, with Mrs of 40–200 kDa (probably the Rh‐related glycoproteins). Under certain experimental conditions glycophorin B appeared to be coprecipitated.

The 2D10 structure, detected by the Mabs, seems to be part of a complex of proteins and/or glycoproteins, which includes Rh polypeptides, the LWab glycoprotein and glycoproteins recognized by various Mabs with Rh‐related specificities. In the red cell membrane, the complex may be associated with glycophorin B.
Detection of residual BCR/ABL transcripts in chronic myeloid leukaemia patients in complete remission using the polymerase chain reaction and nested primers
Authors: Martiat P. , Maisin D. , Philippe M. , Ferrant A. , Michaux J. L. , Cassiman J. J. , Van den Berghe H. , Sokal G.
Published: March 2008

Summary. We sought evidence of BCR/ABL transcripts in the peripheral blood of nine CML patients in complete clinical and cytogenetic remission after treatment by bone marrow transplantation (BMT) or interferon and in one patient who entered spontaneous remission. Six patients were investigated at different times during their follow‐up. We compared results obtained with the polymerase chain reaction (PCR) using (a) a single‐stage PCR comprising 30 cycles of amplification with selected oligomers, and (b) a two‐stage procedure in which the reaction product from the first stage was subjected to a further 30 cycles with nested amplimers. Special care was taken to assess contamination, including for each patient simultaneous co‐extraction of a negative control. Blood cells from all patients showed no evidence of BCR/ABL transcripts in the one‐stage PCR but 9/17 specimens were positive in the two‐stage procedure. Patients in complete remission for a long time (greater than 2 years) appeared negative. These results serve in part to explain the discordant findings reported in other studies and emphasize the importance of carefully selecting the technical conditions most likely to give results that are prognostically relevant for individual patients.
A single nucleotide deletion in codon 123 of the β‐globin gene causes an inclusion body β‐thalassaemia trait: a novel elongated globin chain βMakabe
Authors: Fucharoen Supan , Kobayashi Yukio , Fucharoen Goonnapa , Ohba Yuzo , Miyazono Kohei , Fukumaki Yasuyuki , Takaku Fumimaro
Published: March 2008

Summary. The β‐globin gene from a Japanese individual with an inclusion body β‐thalassaemia trait has been characterized by gene cloning and DNA sequencing. An adenine deletion was detected at the first position of codon 123 (ACC‐CC) of one allele whereas the other allele had a normal sequence. Heterozygosity for this mutation in the patient was confirmed by Southern blots of the genomic DNA digested with HphI, the recognition site of which is eliminated by this deletion. This one base deletion results in the shift of a reading frame in such a manner that the normal termination codon is out of phase. This frameshift mutation results in the synthesis of an elongated β‐globin chain with 10 extra amino acid residues and with an altered C‐terminus. Analysis of labelled globin chains using CM‐cellulose column chromatography failed to demonstrate any abnormal protein, thereby suggesting that the β‐globin chain variant is highly unstable and probably degrades rapidly after synthesis. This event will lead to an accumulation of free α‐chains precipitating in the red blood cells and an inclusion body β‐thalassaemia phenotype would ensue.
Characterization of the erythropoietic progenitor cells (BFU‐E) in paroxysmal nocturnal haemoglobinuria (PNH)
Authors: Terasawa Takashi , Shichishima Tsutomu , Matsuda Minoru , Uchida Tatsumi , Kariyone Shigeo
Published: March 2008

Summary. In order to investigate erythropoiesis in paroxysmal nocturnal haemoglobinuria (PNH), the relative proportions of complement‐mediated lysed erythroblasts in individual bursts of two cases of PNH were analysed by a modified complement lysis sensitivity test using the trypan blue dye exclusion method. All bursts contained both complement‐sensitive and complement‐insensitive erythroblasts and the ratio between them varied from burst to burst. The means ±SD of the haemolytic percentages of individual bursts in cases 1 and 2 were 30.9 ± 10.8% (n= 20, ranging from 13.2% to 52.8%) and 54.8 ± 16.4% (n= 31, ranging from 26.4% to 82.5%), respectively. In addition, the cumulative frequency distributions of the proportions of complement‐mediated lysed erythroblasts in individual bursts approached normal frequency distributions. These results suggest that two red cell populations with different susceptibilities to complement in PNH are derived from a single population of erythropoietic progenitor cells rather than a dual population of normal and PNH erythropoietic progenitor cells.
Acute ‘bilineal‐biphenotypic’ leukaemia
Authors: Akashi Koichi , Shibuya Tsunefumi , Harada Mine , Morioka Eiji , Oshima Koichi , Kimura Nobuhiro , Takeshita Morishige , Kurokawa Mamoru , Kikuchi Masahiro , Niho Yoshiyuki
Published: March 2008

We describe a unique case of hybrid leukaemia with bilineal and biphenotypic features. The coexistence of lymphoblasts and monoblasts was determined morphologically and cytochemically. Immunofluorescence and immunohistochemical analysis revealed that each blast population had both T lymphoid (CD2, cytoplasmic CD3) and myeloid (CD11, CD13, CD15) markers. Southern blot analysis of DNA extracted from the lymph node biopsy demonstrated the presence of monoclonal rearrangement of the TcR‐Cβ gene. Cytogenetic analysis of the bone marrow cells showed a karyotype of 48, XY, 7q + in all of the metaphases examined. These observations are suggestive of a monoclonal origin for these two distinct blast populations.
Light chain restriction analysis of bone marrow plasma cells in patients with MGUS or ‘solitary’ plasmacytoma: diagnostic value and correlation with clinical course
Authors: Bezwoda W. R. , Gordon V. , Bagg A. , Mendelow B.
Published: March 2008

Twenty‐eight patients with a diagnosis of ‘solitary’ plasmacytoma or with gammopathy but with nondiagnostic morphologic examination of the bone marrow were investigated using a short‐term bone marrow culture technique which enriched for the plasma cell fraction. The percentage of monotypic plasma cells in these plasma cell enriched cultures was correlated with the subsequent clinical course. The majority of patients with plasmacytoma and a significant number of those with gammopathy but with otherwise non‐diagnostic investigations were found to have a monoclonal plasma cell component of >20%. There was a significant correlation between the percentage of monoclonal plasma cells as detected by bone marrow culture and subsequent progression to disseminated myeloma. These results indicate that early bone marrow involvement can be detected by means of plasma cell culture prior to morphologic identification of marrow plasmacytosis and that short‐term plasma cell culture distinguishes patients with early, low bulk myeloma from those with monoclonal gammopathy of uncertain significance (MGUS).
Production of granulopoiesis‐stimulating and ‐inhibiting activities by T cells associated with malignant cells in lymphomas
Authors: Piccinni Marie‐Pierre , Jacob Marie‐Christine , Bonnefoix Thierry , Sotto Marie‐France , Pegourie Brigitte , Couderc Pierre , Sotto Jean‐Jacques
Published: March 2008

The possible role of T lymphocytes in the formation of granulomatous reactions seen in certain malignant lymphoid tumours was investigated by measuring the granulopoietic colony‐stimulating activity (CSA) and granulopoietic‐inhibiting activity (IA) produced by stimulated T‐lymphocytes isolated from peripheral blood, spleen and lymph nodes of patients and normal subjects.

Lymph‐node T‐cells from patients with benign lymphoid hyperplasia, B‐cell non‐Hodgkin's lymphoma (B‐NHL), and non‐granulomatous Hodgkin's disease (HD) showed no CSA, but the cells produced [A of 40 ± 23%, 40 ± 24% and 50·5 ± 22·5% respectively. The corresponding cells from patients with HD accompanied by granulomatous reactions produced CSA of 6·85 ± 6·5 u/μl and IA of 23·5 ± 21%.

The presence of a granulomatous reaction in malignant lymphoma was correlated with the stimulation of granulopoiesis in vitro by T lymphocytes associated with malignant cells. A correlation was demonstrated between neutrophilic and eosinophilic colonies obtained in vitro under the influence of CSA‐producing T cells isolated from malignant lymphomas and the neutrophils and eosinophils present in the granuloma. These results showed that tumour‐infiltrating T cells play a role in the presence of granulomatous reactions seen in lymphomas.

Peripheral‐blood T cells from healthy subjects, and from patients with B‐NHL, or with HD unaccompanied by granulocytic reactions produced CSAs of, respectively, 5 ± 0·5 u/μl, 4·8 ± 2·2 u/μl and 5·3 ± 0·4 u/μl, and IAs of 45 ± 18%, 50 ± 5·5% and 50·5 ± 7% respectively. The corresponding values for HD patients with granulocytic reactions were CSA, 17 ± 15·5 u/μl, and IA, 9·5 ± 9%. No correlation was demonstrated between neutrophilic colonies obtained in vitro under the influence of HD blood T cells and neutrophils present in blood. Only one correlation was found: between the percentage of eosinophilic colonies and the number of blood eosinophils. HD blood T cells did not seem to explain completely granulocytic reactions seen in blood.
The C–T substitution in the distal CACCC box of the β‐globin gene promoter is a common cause of silent β thalassaemia in the Italian population
Authors: Ristaldi M. S. , Murru S. , Loudianos G. , Casula L. , Porcu S. , Pigheddu D. , Fanni B. , Sciarratta G. V. , Agosti S. , Parodi M. I. , Leone D. , Camaschella C. , Serra A. , Pirastu M. , Cao A.
Published: March 2008

This paper describes four families of Italian descent in each of which the propositus had the clinical phenotype of thalassaemia intermedia, resulting from the compound heterozygous state for high HbA2β thalassaemia and type I silent β thalassaemia. Direct sequencing on amplified DNA and/or oligonucleotide analysis detected, in all families but one, the compound heterozygous state for codon 39 nonsense mutation and the C‐T substitution at position – 101 in the distal CACCC box of the β‐globin gene promoter (βth–101). Members of these families who are heterozygous for high HbA2β thalassaemia showed the codon 39 nonsense mutation, while those with the clinical phenotype of silent β thalassaemia had the βth–101 mutation. In the remaining family, the propositus and one of his siblings had the compound heterozygous state for a molecularly undefined high HbA2β thalassaemia and the βth–101 mutation in combination with the triple α globin gene arrangement. These patients showed a more severe thalassaemia intermedia like clinical phenotype as compared to those with the same β‐globin genotype and a normal α‐globin gene arrangement. In the families investigated the βth–101 was always associated with haplotype I. A group of patients with thalassaemia intermedia from Southern Italy, either homozygous or heterozygous for haplotype I and in whom previous studies had failed to define the mutation in one of the β thalassaemia globin genes, were screened by oligonucleotide analysis for the presence of the βth–101. Three out of nine were positive. These results indicate that the βth–101 mutation is a common cause of the type I silent β thalassaemia phenotype in the Southern Italian population.
Prevalence of haemochromatosis amongst asymptomatic Australians
Authors: Leggett Barbara A. , Halliday June W. , Brown Nigel N. , Bryant Stewart , Powell Lawrie W.
Published: March 2008

We determined the prevalence of iron overload due to homozygous haemochromatosis in an asymptomatic Australian (predominantly Caucasian) population by surveying 1968 employees of two large corporations. Subjects were screened by measurement of transferrin saturation and serum ferritin concentration and, in all subjects with elevation of both indices, percutaneous liver biopsy was performed to establish whether significant iron overload was present. The prevalence of iron overload due to haemochromatosis in this population was 0·36%. The prevalence rate was not significantly different between males and females, suggesting that this autosomal recessive disease is expressed equally in females given an adequate dietary iron supply. The positive predictive value of a transferrin saturation consistently >45% together with an elevated serum ferritin concentration was 64%. It is concluded that the prevalence of significant iron overload due to homozygous haemochromatosis warranting treatment is approximately 1:300 and that transferrin saturation should be included in existing adult health screening programmes.
Correlation of chromosome abnormalities with laboratory features and clinical course in B‐cell chronic lymphocytic leukaemia
Authors: Oscier D. G. , Stevens J. , Hamblin T. J. , Pickering R. M. , Lambert R. , Fitchett M.
Published: March 2008

141 patients with B‐cell chronic lymphocytic leukaemia (B‐CLL) have been studied for a minimum of 12 months and a maximum of 25 years. 30 of 133 patients (32·5%) had greater than 10% FMC7 positive peripheral blood lymphocytes and 19 of 131 patients (14·5%) had a serum or urinary paraprotein. At presentation 88 patients were stage A0, 18 A1, 18A2, 11B and six C. 44 (31%) had progressive disease and 42 (30%) died during the study period. 63 patients had a normal karyotype, 75 a clonal abnormality and in three no metaphases were obtained. The finding of a complex karyotypic abnormality was significantly associated with λ surface phenotype (P<0·01), and the presence of > 10% FMC7 positive cells (P<0·025), and the presence of a paraprotein (P<0·025). Patients whose leuk‐aemic cells had a complex karyotype and those with stuuctural abnormalities of chromosomes 14 and 6 required treatment earlier than those with a normal karyotype.
Clinicopathological diagnosis and treatment of malignant histiocytosis
Authors: Sonneveld Pieter , Lom Kirsten van , Kappers‐Klunne Mies , Prins M. Eric F. , Abels Johannes
Published: March 2008

Summary. The diagnostic findings of malignant histiocytosis (MH) were analysed in 12 consecutive patients in a single institution. Most patients presented with systemic symptoms and lymphadenopathy (92%), splenomegaly (100%) and hepatomegaly (67%). Neurologic symptoms were present in three patients, while involvement of other organs was present in five patients. The incidence of severe thrombocytopenia was 92%, of anaemia 92% and of leucocytopenia 67%. Serum angiotensin converting enzyme, α1‐antitrypsin and lysozyme were independently increased in 6/9, 3/10 and 1/9 patients respectively. High serum levels of tumour necrosis factor (TNF) were present in 3/10 patients, while serum levels of interleukin‐1 were normal in 10/10 patients. Histologic evidence of MH was obtained in all patients by repeated biopsies of involved tissues. Four patients died prior to treatment. Seven patients were treated with combination chemotherapy, consisting of CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone) or MOPP (chloromethine, vincristine, procarbazine, prednisone), in some cases followed by non‐cross‐resistant second line chemotherapy, if no complete response was attained. The response rate of treated patients was 5 7%, and progression was observed in two patients. The median duration of response was 38 months. Three patients are alive without evidence of disease and off therapy (30+, 83+, 85+ months).

Although MH is a potentially lethal disease, combination chemotherapy may offer a chance for cure in some patients.
Blood transfusion in the para‐Bombay phenotype
Authors: Lin‐Chu M. , Broadberry R. E.
Published: March 2008

Summary. The H‐deficient phenotypes found in Chinese so far, have all been secretors of soluble blood group substances in saliva. The corresponding isoagglutinin activity (e.g. anti‐B in OBHm persons) has been found to be weak in all cases. To determine the clinical significance of these weak isoagglutinins 51Cr red cell survival tests were performed on three OBHm individuals transfused with small volumes (4 ml) of groups B and O RBC. Rapid destruction of most of the RBC occurred whether or not the isoagglutinins of the OBHm individuals were indirect antiglobulin test (IAGT) reactive. When a larger volume (54 ml packed RBC) of group B cells (weakly incompatible by IAGT) was transfused to another OBHm individual with IAGT active anti‐HI, the survival of the transfused RBC was 93% at 24 h, with 30% of the RBC remaining in the circulation at 28 d in contrast to 76% as would be expected if the survival was normal. Therefore when whole units of blood of normal ABO blood groups, compatible by IAGT, are transfused, the survival is expected to be almost normal. These weak isoagglutinins may not be very clinically significant and we suggest that when para‐Bombay blood is not available, the compatibility testing for OAHm persons should be performed with group A and group OBHm packed RBC; OBHm with group B and group O packed RBC; OABHm with groups A, B, AB and O packed RBC. For cross matching, the indirect antiglobulin test by a prewarmed technique should be used.
Interleukin‐4 regulates mRNA accumulation of macrophage‐colony stimulating factor by fibroblasts: synergism with interleukin‐1β
Authors: Henschler Reinhard , Mantovani Luisa , Oster Wolfgang , Lübbert Michael , Lindemann Albrecht , Mertelsmann Roland , Herrmann Friedhelm
Published: March 2008

Summary. We demonstrate that macrophage‐colony stimulating factor (M‐CSF) is expressed in human fibroblasts at the mRNA and protein level. Following activation with both interleukin (IL)‐1β and IL‐4. fibroblasts synthesized M‐CSF transcripts detectable by Northern blot analysis with peak expression occurring at 8 h and 12 h, respectively. Exposure of fibroblasts to both cytokines resulted in M‐CSF protein release at 60 h of c. 500 U/ml (for IL‐1β) and 1000 U/ml (for IL‐4), relative to a control preparation of recombinant human M‐CSF in a murine bone marrow colony assay. Both interleukins synergized to enhance M‐CSF mRNA accumulation and their ability to induce M‐CSF transcripts could be abolished by treatment with specific neutralizing antibodies. These observations provide support for the idea that fibroblasts may control monocyte/macrophage development and function, and that IL‐1β and IL‐4 are involved in the regulation of this process.
Translocation of bcl‐2 gene in non‐Hodgkin's lymphomas in Hong Kong Chinese
Authors: Loke S. L. , Pittaluga S. , Srivastava G. , Raffeld M. , Ho F. C. S.
Published: March 2008

Summary. The close association between translocation of the proto‐oncogene bcl‐2 and follicular lymphomas has been well established in Caucasian patients and the de‐regulation of bcl‐2 has been implicated in follicular lymphomagenesis. Similar molecular structural alterations have also been detected in diffuse lymphomas with a previous history of a follicular pattern as well as in a smaller proportion of de novo diffuse lymphomas.

There is a lower incidence of follicular lymphomas in Chinese. In order to investigate further this phenomenon, we used bcl‐2 translocation as a genetic marker of follicular lymphomas, to study 31 cases of B cell non‐Hodgkin's lymphomas in Chinese patients by Southern blot analysis.

Eight out of 16 cases of follicular lymphomas showed bcl‐2 translocation with involvement of the major breakpoint region (MBR). Six of these cases utilized breakpoints within the 4.3 kb HindIII fragment, while in two cases the breakpoints were more dispersed, but still within the BamHI fragment. An additional case of follicular lymphoma showed translocation of bcl‐2 gene with involvement of the minor cluster region (mcr), making a total of nine out of 16. None of the 15 cases of diffuse lymphomas showed similar molecular structural alterations.

These data show that bcl‐2 translocation is present in 57% of follicular lymphomas in Chinese patients, and support the notion that bcl‐2 translocation is a consistent marker for follicular lymphomas irrespective of ethnic differences. As the translocation is not detected in the diffuse lymphomas, there is no evidence to suggest that the low incidence of follicular lymphomas in Chinese patients is due to a greater tendency for follicular tumours to progress rapidly and present as diffuse lymphomas.
Capacity to produce interleukin 2 is impaired in haemophilia in the absence and presence of HIV 1 infection
Authors: Madhok R. , Gracie J. A. , Smith J. , Lowe G. D. O. , Forbes C. D.
Published: March 2008

Summary. Capacity to produce interleukin‐2 (IL‐2) was measured in haemophiliacs from a well‐defined treated cohort. Patients were selected on the basis of HIV‐1 antibody status, mean annual dose of clotting factor and liver disease severity. T‐cell subsets and peripheral blood mononuclear cell proliferation to Mycobacterium tuberculosis purified protein derivative (PPD) were also measured.

Haemophiliacs had reduced IL‐2 production, independent of HIV‐1 antibody status, mean annual dose of clotting factor concentrate used and liver disease severity. In HIV‐1 antibody positive patients reduced levels correlated with PPD proliferative responses (r=0.6, P=0.04) and CD8+ve (r=05, P=0.05) but not CD4 + ve cell numbers (r= 0.3, P=0.2). No such correlations were seen in HIV‐1 antibody negative patients.

Reduced IL‐2 production in HIV‐1 antibody negative haemophiliacs was due to a qualitative defect. In HTV‐1 positive patients a qualitative defect in T lymphocytes that selectively proliferate in response to PPD was observed. CD4 + ve cell numbers were reduced in HIV‐1 positive patients.
Inhibition of activated protein C and its cofactor protein S by antiphospholipid antibodies
Authors: Malia R. G. , Kitchen S. , Greaves M. , Preston F. E.
Published: March 2008

Summary. We have investigated the effects of purified IgG fractions from plasma containing the lupus anticoagulant (LAC) and/or IgG anticardiolipin antibody (ACA) on the degradation of factor Va by an activated protein C‐protein S complex. Plasma samples from 10 patients were studied. LAC was detected by a Russell's Viper venom technique. ACA was determined by ELISA. IgG fractions were obtained from each plasma sample by protein A‐Sepharose fractionation. This fraction was shown to exhibit ACAILAC activity. Using purified activated protein C (APC), protein S and phosphati‐dylserine/phosphatidylcholine, factor Va degradation was assessed in the presence and absence of IgG fractions from LAC/ACA containing plasmas. After 2 min incubation the mean factor Va degradation by APC and protein S in the presence of IgG LAC/ACA fractions was 14% compared with 52% with normal IgG. A similar effect was seen when phospholipid was substituted by washed freeze‐thawed platelets. Experiments employing varying concentrations of protein S and phospholipid revealed marked differences in respect of the inhibitory specificity of the different antiphospholipid antibodies. These results indicate that antiphospholipid antibodies have an inhibitory effect on the activated protein C/protein S complex and provide some explanation for a relationship between antiphospholipid antibodies and thrombosis.
Clonal karyotype abnormalities and clinical progress in the myelodysplastic syndrome
Authors: Geddes A. D. , Bowen D. T. , Jacobs A.
Published: March 2008

Summary Summary Clonal karyotype abnormalities in 124 patients with myelodysplastic syndrome are reviewed. 36% of patients had abnormalities at referral, the most common being 5q ‐, +8 and lesions of chromosomes 7 and 20. Reduced survival was associated with the presence of either single or multiple clonal abnormalities at referral, abnormalities of chromosome 7 or 8 (either alone or with other lesions) and exclusively abnormal metaphases. The presence of 5q ‐alone did not appear to affect survival.

Sequential studies were carried out in 77 patients of whom 12 showed karyotypic evolution. Reduced survival was observed in patients with an evolving karyotype but appeared to be due almost entirely to evolution in those patients whose initial karyotype was normal. Leukaemic transformation occurred more commonly in patients with an abnormal karyotype, particularly those with multiple abnormalities, and in patients with an evolving karyotype. Although the first appearance of an abnormal karyotype or an apparent evolution are important phenomena, it is probable that in some cases they merely represent expansion of a previously existing clone that has escaped detection. The distinction between true karyotypic evolution or clonal expansion and statistical variations due to small sample size and variability of samples may be difficult but needs to be taken into account in considering clinical significance.
Expression and modulation of IL‐3 and GM‐CSF receptors in human growth factor dependent leukaemic cells
Authors: Brizzi M. F. , Avanzi G. C. , Veglia F. , Clark S. C. , Pegoraro L.
Published: March 2008

Summary From the interleukin‐3 (IL‐3) responsive human myeloid cell line M‐07 we derived a subclone, named M‐07e. which is fully dependent for growth and survival on either granulocyte‐macrophage colony stimulating factor (GM‐CSF) or IL‐3. In this paper the expression, specificity and modulation of GM‐CSF and IL‐3 receptors on M‐07e cells are described. The specificity of the binding was demonstrated by the failure of other cytokines to compete, at 4°C. with GM‐CSF or IL‐3 receptors. In addition, IL‐3 was found to compete as well as GM‐CSF for GM‐CSF receptors while GM‐CSF was a weak competitor for IL‐3 receptors. Quantitative binding studies and Scatchard plot analysis revealed the presence of a single class of high‐affinity GM‐CSF binding sites (405 ± 2 74 sites per cell, dissociation constant at the equilibrium 52 ± 20 PM) and the presence of high and low‐affinity regions for IL‐3 binding sites (2 7 ± 12 and 416 ± 92 sites per cell for the high and low affinity regions respectively, dissociation constant at the equilibrium, 58 ± 22 PM and 5.7 ± 2.0 nM respectively). Finally, in agreement with the hierarchical down‐modulation model, we found that IL‐3 can down‐modulate both IL‐3 and GM‐CSF receptors while GM‐CSF can down‐modulate only its own receptors. The present results suggest that M‐07e cells, in spite of their neoplastic nature, share, with murine bone marrow cells, similar growth factor receptor regulatory mechanisms. This cell line may be regarded as a candidate model to investigate the physiological events triggered by growth factors binding to human haemopoietic cells.
Histocompatibility antigen patterns in haemophilic patients with factor VIII antibodies
Authors: Aly Ashraf M. , Aledort Louis M. , Lee Tsung D. , Hoyer Leon W.
Published: March 2008

Summary A number of studies suggest that there is a genetic basis for the formation by some haemophilia A patients of antibodies that inactivate factor VIII. In our study, human leucocyte antigen (HLA)‐A, B. C, DR and DQ typing was carried out for 44 haemophilia A patients, including 16 who had developed an antibody to factor VIII. In contrast to previous reports, we found no association between HLA‐DR antigens and haemophilia A per se or the formation of a factor VIII inhibitor. However, there was an absence of HLA‐Cw5 in the 16 haemophilic patients who had formed an antibody to factor VIII. This finding, consistent with a previous report, identified a statistically significant difference in HLA‐Cw5 frequency when the inhibitor patient group was compared to multi‐transfused haemophilic patients who had no inhibitor (11/28).
Basal and induced prothrombinase expression in platelets from patients with essential thrombocythaemia (ET)
Authors: Crook M. , Machin S. J. , Crawford N.
Published: March 2008

Summary Both the basal and the collagen plus thrombin‐stimulated prothrombinase activities have been measured in platelets from eight patients with essential thrombocythaemia (ET). On a mean basis these activities were significantly lower than the values recorded for a group of age and sex matched control subjects. When the Ca2+ ionophore A23187 was substituted for the agonist mixture the induced expression of prothrombinase activity was essentially the same for the patient and control groups. It is suggested that the defect in prothrombinase expression seen with ET platelets may reside in either membrane signal transduction processes concerned with Ca2+ mobilization or in Ca2+ sensitive cytoskeletal control of membrane phospholipid topography.
Selection of a panel of monoclonal antibodies for monitoring residual disease in peripheral blood and bone marrow of interferon‐treated hairy cell leukaemia patients
Authors: Palini B. , Pileri S. A. , Flenghi L. , Liberati M. , Stein H. , Gerli R. , Minelli O. , Martelli M. F. , Lauria F. , Poggi S.
Published: March 2008

Summary A panel of monoclonal antibodies (mAbs) directed against B‐cell and hairy cell leukaemia (HCL)‐associated antigens was used to identify residual hairy cells in the peripheral blood and/or bone marrow samples from 20 patients with HCL. following treatment with interferon‐alpha (IFN‐alpha) or interferon‐beta (IFN‐beta). In all cases, hairy cells retained their characteristic phenotype, e.g. positivity for CD22, CD11c, CD25, CD32, and the HCL‐associated trimeric protein (t‐GP) recognized by the mAbs HML‐1, B‐ly7, LF61 and Ber‐Act8. The most specific marker for identifying a small percentage of hairy cells in peripheral blood cytospins. was t‐GP. In alkaline phosphatase/anti alkaline phosphatase (APAAP) stained preparations, t‐GP+ hairy cells (provided with large cytoplasm and hairy surface) could be usually distinguished from t‐GP+ normal lymphocytes (small‐sized cells with smooth surface). In doubtful cases the percentage of residual hairy cells could exactly be estimated by double immunofluorescence staining for CD22 (B‐cell marker) and t‐GP. The rationale of the test is based on the finding that the small percentage (about 1%) of t‐GP + lymphocytes circulating in the peripheral blood of normal individuals are T‐cells of the CD8 subset and not B‐cells. The best markers for identifying residual hairy cells in routine bone marrow biopsies were CD45RA (mAb 4KB5) and CD20 (mAb L26). Immunohistological labelling was superior to morphological examination in picking up scattered hairy cells in bone marrow biopsies showing either severe hypoplasia or exuberant hyperplasia of normal haemopoietic series.