bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cryopreservation disturbs stimulus‐response coupling in a platelet subpopulation
Authors: Dullemond‐Westland A. C. , Van Prooijen H. C. , Rlemens M. I. , Akkerman J. W. N. 
Published: July 2010

SummaryCryopreservation of platelet suspensions leads to a severe reduction in the functional responses of these cells. This detect primanly resides in a subpopulation which consists of about half the total platelet mass and shows an increased susceptibility to the Cryopreservation procedure. Following freeze‐thawing this subpopulation failed to aggregate upon stimulation with collagen despite a normal appearance of membrane glycoproteins. Also the contents of α granules was similar to the whole suspension and since changes in α and dense granule contents often occur in parallel, the poor aggregability could not be attributed to a defect in secretion granules. In contrast, the generation of phosphatidyl‐inositol metabolites, the increase in cytosolic Ca2+ content and the phosphorylation of proteins of mol wt 47 000 and 20 000 seen when normal platelets are activated, were completely absent in the subpopulation. These data reveal a severe defect in stimulus‐response coupling mechanisms in about half of the platelets after freeze‐thawing. The other half of the suspension endures this treatment relatively unharmed and may be used for transfusion.
Glycolipids and glycopeptides of red cell membranes in congenital dyserythropoietic anaemia type II (CDA II)
Authors: Zdebska E. , Anselstetter V. , Pacuszka T. , Krauze R. , Chelstowska A. , Heimpel H. , Kosacielak J. 
Published: July 2010

SummaryThe composition and structure of neutral and acidic oligoglycosylceramides, polyglycosylceramides and polyglycosylpeptides were determined in erythrocyte membranes of two patients with congenital dyserythropoietic anaemia type II. In keeping with previous studies we found an elevated accumulation in CDA II erythrocytes of LacCer, Lc3Cer and nLc4Cer. Gb4Cer was elevated in erythrocytes of only one of the two patients tested. In addition we found a significant increase of 6IVNeuAcnLc4Cer ganglioside. Polyglycosylceramides were elevated 6‐fold but they resembled those of cord erythrocytes with respect to complexity and the number of side chains. Polyglycosylpeptides of CDA II erythrocytes were decreased 7‐fold. These glycopeptides were, however, heterogeneous with respect to branching pattern; the minor fraction was highly branched whereas the major one was more linear in structure. Both polyglycosylceramides and polyglycosylpeptides exhibited high I and i antigenicity. We postulate that the accumulation of glycolipids and underglycosylation of glycoproteins in CDA II membranes results from the prolongation of G1 and possibly M phases of the mitotic cycle of the erythroid cells in which glycolipids are preferentially synthesized.
Platelet storage pool deficiency: diagnosis in patients with prolonged bleeding times and normal platelet aggregation
Authors: Israels S. J. , McNicol A. , Robertson C. , Gerrard J. M. 
Published: July 2010

SummaryWe evaluated 46 patients with prolonged bleeding times, and no demonstrable abnormalities of either von Willebrand factor or platelet aggregation, for possible deficiency of platelet storage pool. Studies of ATP release from thrombin‐stimulated platelets and enumeration of dense granules in platelet whole mounts were performed in these patients. Seventeen patients (35%) had both decreased ATP release and decreased numbers of dense granules, suggesting the presence of a storage pool defect. Thus, storage pool deficiency may be present in the absence of the classical aggregation abnormalities. Evidence of storage pool deficiency should be considered in all patients with an isolated unexplained prolongation of the bleeding time. The methods used in this study are readily applicable to most clinical laboratories.
Arachidonic acid metabolism in platelets stored for 5 days
Authors: Cesar J. M. , Navarro J. L. 
Published: July 2010

Summary(C‐14)‐Arachidonic acid [(C‐14)‐AA] metabolism was studied in platelet concentrates (PCs) stored for 5 d. There was a gradual decrease in uptake of radioactivity from day 0 to 3 (P < 0.01). On day 0, distribution of radioactivity in platelet phospholipids (PLs), and formation of phosphatidic acid, HETE and cyclooxygenase products, when platelets were exposed to thrombin (5 U/ml), were similar to that reported for fresh platelets. On day 3 there was a change in the distribution of (C‐14)‐AA in platelet PLs which consisted of an increase in the percentage of radioactivity bound to phosphatidylserine, from 5.3±0.9% on day 0 to 8.8±1.5% on day 3 (P<0.001), and a decrease in (C‐14)‐AA in phosphatidylinositol (PI), from 12.4±1.5% on day 0, to 7.9±0.9% on day 3 (P<0.001). Phosphatidic acid generated by thrombin‐stimulated platelets on day 0, comprised 2.6±0.5% of total radioactivity, but dropped to 1.4±0.3% on day 3 (P<0.001), and 0.9±0.2% on day 5 (P<0.01). These values showed a good correlation with the percentage of (C‐14)‐AA released from PI on the same days (r=0‐9). On day 0, 13.4 ± 4.4% of platelet radioactivity was released from phosphatidylcholine by thrombin, but this amount was reduced to 6.8±3.4% on day 5 (P<0.05). Generation of radioactive 12‐hydroxy‐5,8,10,14‐eicosatetraynoic acid (HETE) also dropped from 7.2±2.9 on day 0. to 2.1 ± 1 on day 5 (P< 0.01). We could not detect changes in cyclooxygenase metabolites. In conclusion, we suggest that various enzymatic pathways implicated in AA metabolism by platelets are impaired by storage.
Cytogenetic studies in splenic lymphoma with villous lymphocytes
Authors: Oscier D. G. , Matutes E. , Gardiner A. , Glide S. , Mould S. , Brito‐Babapulle V. , Ellis J. , Catovsky D. 
Published: July 2010

Summary. We report the cytogenetic findings on 31 cases of splenic lymphoma with villous lymphocytes (SLVL). TPA stimulated cells from peripheral blood (28 cases), spleen (two cases) and lymph node (one case) with SLVL have been analysed. A clonal chromosome abnormality was found in 27/31 patients (87%): this was identified as a simple abnormality in 12 cases and a complex one in 15. Four recurring abnormalities were seen: t(11:14) (q13:q32)in five patients, deletions or translocations involving 7q in seven patients. iso 17q in four patients and translocations involving 2p11 in four patients. The high frequency of clonal chromosome abnormalities in SLVL contrasts with the usually benign clinical course of this disease.Abnormalities found frequently in patients with chronic lymphocytic leukaemia (CLL) such as trisosmy 12 and deletions or translocations involving 13q14 were each seen in only one patient. No case had the t(14:18) characteristic of follicular lymphoma.Our findings demonstrate the high frequency of clonal and often complex chromosome abnormalities in SLVL. Although a unique chromosome rearrangement has not been identified, a pattern of four recurrent abnormalities has emerged. Our results suggest that SLVL is distinct on cytogenetic grounds from B‐CLL and follicular lymphoma but shows similarity with mantle cell lymphoma, lymphoplasmacytic lymphoma and B‐PLL.
Cytokine gene expression in human multiple myeloma
Authors: Portier Marielle , Zhang Xue‐Guang , Ursule Elisabeth , Lees Delphine , Jourdan Michel , Bataille Regis , Klein Bernard 
Published: July 2010

Summary. In the present study the gene expression of cytokines promoting in vitro myeloma‐cell growth was investigated by Northern blot analysis using total RNA of 36 tumour samples of patients with multiple myeloma (MM) or plasma cell leukaemia and poly(A)+ RNA of 10 human myeloma cell lines (HMCL). These cytokines included interleukin (IL)‐1α, IL‐1β, IL‐3, IL‐6, granulocyte‐macrophage (GM)‐colony‐stimulating factor (CSF) and granulocyte (G)‐CSF. IL‐1β, IL‐6 and G‐CSF genes were coexpressed in most patients, although at variable levels. IL‐1α transcripts were detected in 32% of patients in whom coexpression of IL‐1β gene was found. IL‐3 gene was not expressed in patients' cells and GM‐CSF mRNA was detected in only 1/32 patients. No detectable transcripts for the above cytokines were present in HMCL, whereas IL‐6 gene was expressed in 2/10 HMCL.We also looked for the presence of transcripts for IL‐2, leukaemia inhibitory factor (LIF) and transforming growth factor (TGF)β in cells of tumour samples from the same patients and in HMCL. IL‐2 gene was not expressed in MM patients and HMCL. Weak expression of LIF gene was detected in three patients (9%), and transforming growth factor β (TGFβ) mRNA was observed in 12/12 tumour samples analysed and all HMCL.These results suggest that, among cytokines shown to control myeloma‐cell growth in vitro, IL‐1, IL‐6 and G‐CSF could play a role in the development of myeloma disease in vivo.
In vitro inhibition of tumour necrosis factor‐α and interleukin‐6 production by intravenous immunoglobulins
Authors: Toungouz Michel , Denys Chantal H. , Groote Donat De , Dupont Etienne 
Published: July 2010

In vitro data about the action of pooled immunoglobulins (Ig) on cytokine (CK) production are controversial. The recent finding of natural antibodies against staphylococcal toxins neutralizing superantigen‐induced activation prompted us to design an assay determining their ability to modulate staphylococcal enterotoxin B (SEB) induced CK production (IL‐6 and TNF‐α). Presence of anti‐SEB antibodies was demonstrated by a dot‐blot assay in the three preparations tested. Preincubation of SEB with pooled Ig prior to addition into the test tube containing PBMCs (neutralizing condition) resulted in a strong inhibition of both TNF‐α and IL‐6 release (TNFα: 59 ± 5% inhibition, P<0.0001; IL‐6: 71 ± 7% inhibition. P<0.0001, n= 15). Anti‐CD3 MoAb‐induced CK production was not modified. During our study it was found that experimental conditions were critical to observe this inhibitory effect. Reversing the previous procedure by adding PBMCs into the test tube containing pooled Ig mixed with SEB resulted in a marked induction of TNF‐α and IL‐6 production. The same observation was made when pooled Ig solely was added (coating condition). F(ab')2 fragments of pooled Ig displayed similar inhibitory capacity when added in neutralizing condition, indicating that the mechanism involved was not Fc dependent. The fragments lost the activating properties of intact Ig when incubated in coating condition, showing that Fc receptor activation occurs in this setting.The present work demonstrates that inhibition of SEB‐induced CKs release by pooled Ig can be achieved by SEB neutralization, provided that the experimental conditions avoid activation through the Fc receptor. It can be assumed that similar mechanisms take place in some clinical conditions during which pooled Ig are infused.
Establishment of a myeloid leukaemic cell line (SKNO‐1) from a patient with t(8;21) who acquired monosomy 17 during disease progression
Authors: Matozaki Sachiko , Nakagawa Toshitaro , Kawaguchi Ryuji , Aozaki Ryoji , Tsutsumi Masayoshi , Murayama Tohru , Koizumi Tamio , Nishimura Ryuichiro , Isobe Takashi , Chihara Kazuo 
Published: July 2010

A novel cell line SKNO‐1 was established from the bone marrow cells of a 22‐year‐old male suffering from acute myeloblastic leukaemia (AML) M2 with t(8;21) whose disease became resistant to chemotherapy after acquisition of 17 monosomy. SKNO‐1 has been maintained for more than 36 months as a granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) dependent line.Morphologically, SKNO‐1 cells were myeloblasts somewhat maturated. The cells grow in suspension with a doubling time of 48‐72 h. The survival and growth of SKNO‐1 cells was absolutely dependent on granulocyte‐macrophage colony stimulating factor (GM‐CSF). SKNO‐1 cells possessed t(8;21) and monosomy 17 which were observed in original leukaemic cells. We confirmed that the AML1 gene, located on chromosome 21, was rearranged and the AML1‐MTG8 fusion transcript was expressed in SKNO‐1 cells. Over‐expression and mutation of the p53 gene were also detected in SKNO‐1. It is likely that alterations of AML1 or MTG8 gene and p53 gene contribute to a disease progression in this case.Since t(8;21) translocation is a common chromosome abnormality in AML, and inactivation of the p53 gene may play a crucial role in disease progression in AML, SKNO‐1 would be a useful tool for analysing the molecular mechanisms in myeloid leukaemogenesis.
Primitive haematopoietic progenitors in the blood of patients with sickle cell disease appear to be endogenously mobilized
Authors: Croizat H. , Ponchio L. , Nicolini F. E. , Nagel R. L. , Eaves C. J. 
Published: July 2010

To investigate whether haematopoietic stem cells in patients with sickle cell (SS) disease might be altered, we examined the number and cycling status of 5‐week long‐term culture‐initiating cells (LTC‐ICs) and in vitro multilineage colony‐forming cells (CFCs) present in the blood of a large and clinically diverse group of SS patients. The concentrations of both of these cell types per ml of blood varied over a wide range in individual patients, but, on average, were significantly elevated above normal values (∼sevenfold and 15‐fold respectively) and to an even greater extent than the lineage‐restricted CFCs in the same samples. Wide variations in the concentration of circulating progenitors, particularly the LTC‐ICs, were also seen over time (in concert with changes in the white blood cell count) in SS patients. [3H]‐Thymidine suicide assays showed most of the CFCs and LTC‐ICs in SS blood to be quiescent like their counterparts in normal blood. However, by comparison with historical data, the SS progenitors could be recruited into the cycle more quickly (i.e. within 2 vs. 3 d), thus showing the same kinetics of activation exhibited by ‘mobilized’ progenitors from patients given chemotherapy and exogenous growth factors. Taken together, these findings implicate previously documented increases in endogenous Steel factor, interleukin 3 and granulocyte–macrophage colony‐stimulating factor levels in SS patients in the establishment of a chronically mobilized progenitor phenotype.
Quality of harvested autologous platelets compared with stored donor platelets for use after cardiopulmonary bypass procedures
Authors: Crowther M. , Ford I. , Jeffrey R. R. , Urbaniak S. J. , Greaves M. 
Published: July 2010

Platelet dysfunction has a major contribution in bleeding after cardiopulmonary bypass (CPB) and transfusion of platelets is frequently used to secure haemostasis. Allogeneic platelets prepared for transfusion are functionally impaired. Autologous platelets harvested preoperatively require a shorter storage time before transfusion and their use also avoids the risks associated with transfusion of allogeneic blood products. For the first time, we have compared the functional quality of autologous platelets with allogeneic platelets prepared by two methods, immediately before infusion. Platelet activation was assessed by P‐selectin expression and fibrinogen binding using flow cytometry. We also monitored the effects of CPB surgery and re‐infusion of autologous platelets on platelet function. Autologous platelet‐rich plasma (PRP) contained a significantly lower (P < 0·05) percentage of P‐selectin‐positive and fibrinogen‐positive platelets compared with allogeneic platelet preparations, and also contained a significantly higher (P < 0·05) percentage of responsive platelets. Allogeneic platelets prepared by donor apheresis were more activated and less responsive than those produced by centrifugation of whole blood. In patients' blood, the percentage of platelets expressing P‐selectin or binding fibrinogen increased significantly after CPB (P < 0·05), while the percentage of platelets responsive to in vitro agonists was decreased (P < 0·05 in autologous transfusion patients), consistent with platelet activation during the procedure. The percentage of activated platelets decreased (statistically not significant) after re‐infusion of autologous PRP. P‐selectin expression had returned to pre‐CPB levels 24 h post‐operatively. Autologous platelet preparations display minimal activation, but remain responsive. Conservation of platelet function may contribute to the potential clinical benefits of autologous transfusion in cardiopulmonary bypass.
Extended routine polymerase chain reaction surveillance and pre‐emptive antiviral therapy for cytomegalovirus after allogeneic transplantation
Authors: Peggs Karl S. , Preiser Wolfgang , Kottaridis Panagiotis D. , McKeag Nikki , Brink Nicola S. , Tedder Richard S. , Goldstone Anthony H. , Linch David C. , Mackinnon Stephen 
Published: July 2010

Pre‐emptive treatment strategies based on sensitive screening for cytomegalovirus (CMV) infection up to day +100 after allogeneic transplantation have been shown to reduce the incidence of CMV disease during the period of surveillance. However, the use of ganciclovir has been associated with delays in immune reconstitution and an increased incidence of late CMV disease after day +100. In the present study, 81 patients undergoing allogeneic transplantation received polymerase chain reaction (PCR)‐guided pre‐emptive therapy based on detection of CMV DNA by PCR on 2 consecutive weeks up to day +180. Thirty‐three of the 52 high‐risk patients (CMV‐seropositive donor or recipient) received a total of 45 treatment episodes up to day +100. Three of these patients (5·7%) developed CMV disease, with one fatality. Twelve of the surviving 44 high‐risk patients (27%) required pre‐emptive treatment between days +101 and +192, but none of these patients developed late CMV disease with a median follow‐up of 402 d (range 117–952 d). Antiviral therapy was stopped after a single negative PCR result with no subsequent episodes of CMV disease while patients remained off antiviral treatment. As all initial episodes of CMV DNA detection occurred within 60 d of transplantation, it may be possible to discontinue monitoring beyond day +100 in patients who have remained CMV PCR negative before this. Thus, we have confirmed that PCR‐guided pre‐emptive therapy results in a low incidence of CMV disease before day +100 and that discontinuing treatment on the basis of viral clearance as determined by CMV PCR appears to be safe practice. In addition, we have observed no episodes of late CMV disease with an extension of surveillance to 26 weeks.
Phenotypic and molecular diversity of haemoglobin H disease: a Greek experience
Authors: Kanavakis Emmanuel , Papassotiriou Ioannis , Karagiorga Markissia , Vrettou Christina , Metaxotou‐Mavrommati Anna , Stamoulakatou Alexandra , Kattamis Christos , Traeger‐Synodinos Joanne 
Published: July 2010

Haemoglobin H (Hb H) disease is the severest form of α‐thalassaemia compatible with post‐natal life and occurs when α‐thalassaemia mutations interact to reduce α‐globin synthesis to levels approximately equivalent to the output of a single α‐globin gene. Hb H disease has variable clinical expression, mainly related to underlying genotypes. The spectrum of α‐thalassaemia determinants in Greece appears greater than in any other population studied and, in 75 Greek Hb H disease patients, we found 12 α‐thalassaemia mutations interacting to produce 15 Hb H disease genotypes. Evaluation of haematological, biochemical and clinical findings, and correlation with genotypes, defined genetic predictors of disease severity and factors involved in disease progression. In accordance with previous reports, patients with non‐deletion α‐thalassaemia mutations had more severe clinical expression. Additionally, we found that all patients with the most severe phenotypes had α‐thalassaemic globin variants. Phenotypic severity was not simply related to the degree of α‐globin deficiency: high Hb H levels were found to exacerbate anaemia by negatively influencing tissue oxygenation, and both Hb H and α‐thalassaemic haemoglobin variants appear to reduce red cell survival within the bone marrow and circulation. Together with the long‐term follow‐up in many patients, this report provides comprehensive information for management of Hb H disease and appropriate family counselling.
RUNX1 gene mutation in primary myelodysplastic syndrome – the mutation can be detected early at diagnosis or acquired during disease progression and is associated with poor outcome
Authors: Chen Chien‐Yuan , Lin Liang‐In , Tang Jih‐Luh , Ko Bo‐Sheng , Tsay Woei , Chou Wen‐Chien , Yao Ming , Wu Shang‐Ju , Tseng Mei‐Hsuan , Tien Hwei‐Fang 
Published: July 2010

SummaryMutations of Runt‐related transcription factor 1 (RUNX1) have been detected in patients with myelodysplastic syndrome (MDS). However, the prognostic implication of RUNX1 mutations in primary MDS is limited. The stage of the disease at which the mutations are acquired and whether they persist during the disease course also remain unclear. We analysed mutations of RUNX1 exons 3–8 in 132 patients with primary MDS and correlated the results with clinical features. Serial studies were performed during the follow‐up period. Sixteen patients (12%) had RUNX1 mutations at the time of diagnosis. All RUNX1 mutations that were detected at diagnosis remained unchanged during the clinical course. Two other patients acquired RUNX1 mutations at leukaemic transformation 34 months and 35 months after the diagnosis of MDS. Patients with RUNX1 mutations at diagnosis had higher neutrophil counts and higher frequency of −7/7q deletion than those without. Furthermore, RUNX1 mutation was closely associated with a short overall survival (P = 0·039). This is the first report to demonstrate that RUNX1 mutation can not only be detected early at diagnosis but also acquired during disease progression and is associated with poor prognosis in patients with primary MDS. It may play a role in the development and progression of a subset of primary MDS.
Ciclosporin and plasma exchange in thrombotic thrombocytopenic purpura: long‐term follow‐up with serial analysis of ADAMTS13 activity
Authors: Cataland Spero R. , Jin Ming , Lin Shili , Kennedy Melanie S. , Kraut Eric H. , George James N. , Wu Haifeng M. 
Published: July 2010

SummaryWe hypothesized that ciclosporin (CSA) as adjunct to plasma exchange (PE) improves the efficacy of PE in idiopathic thrombotic thrombocytopenic purpura (TTP) via suppression of the antibody inhibitor of ADAMTS13. Our preliminary findings with CSA and PE as the upfront treatment of TTP suggested that the addition of CSA to PE significantly decreased the exacerbation (disease recurrence within 30 d of the last PE) rates compared to a cohort that received corticosteroids and PE as their upfront therapy of TTP. We present an updated analysis with long‐term follow‐up of 18 patients with idiopathic TTP treated with concurrent CSA and PE with analysis of serial measurements of ADAMTS13 activity, antigen and inhibitor concentration in the context of clinical outcome data. Overall, 16/18 (89%) patients achieved remission, similar to historical remission rates in idiopathic TTP with PE with only one patient suffering an exacerbation. Clinical responses correlated with improvements in ADAMTS13 activity and suppression of the antibody inhibitor of ADAMTS13. These data suggest that the efficacy of CSA is at least in part related to its suppression of the antibody inhibitor of ADAMTS13 and a subsequent improvement in ADAMTS13 activity and antigen.