bluuebunny/crossref_metadata_2025 huggingface dataset:
Spouses of demented patients with low cobalamin levels: a new risk group for cobalamin deficiency
Authors: Carmel R. , Cairo K. , Bondareff W. , Gott P. S. , Cummings J. L. , Henderson V. W.
Published: July 2010

Abstract: Low serum cobalamin levels are common in conditions such as dementia and often represent mild deficiency. We surveyed serum cobalamin levels prospectively in spouses and blood relatives of demented patients to determine if any familial predisposition exists for the low levels. Cobalamin status in most of the relatives found to have low levels was assessed further by means of blood counts, metabolic tests, neurologic evaluation, absorption studies and response to cobalamin therapy. Serum cobalamin levels in 36 spouses correlated with those of the 36 demented patients related to them (r = 0.46, p = 0.004). A significant association was not seen in 34 blood relatives of 34 demented patients (r = 0.27). Most importantly, 67% of the spouses of demented patients with low serum cobalamin had low values themselves, compared with only 3% of the spouses of patients with normal levels (p = 0.001). Detailed study of 4 of the 5 spouses (and 3 blood relatives) with low cobalamin levels showed no anemia in any case. Nevertheless, 4 of the subjects had metabolic evidence of deficiency and one had electrophysiological abnormalities; all these defects improved with cobalamin therapy. These observations identify a hitherto unsuspected group of people at high risk for cobalamin deficiency and suggest that spouses of demented patients with low cobalamin levels should also have their cobalamin levels measured. The increased frequency of low serum cobalamin levels in spouses of demented patients with low levels represents in most cases a true, mild cobalamin deficiency that responds to treatment.
Serum thrombopoietin and plasma glycocalicin concentrations as useful diagnostic markers in thrombocytopenic disorders
Authors: Kunishima S. , Tahara T. , Kato T. , Kobayashi S. , Saito H. , Naoe T.
Published: July 2010

Abstract: Using enzyme‐linked immunosorbent assays, we measured the concentrations of serum thrombopoietin (TPO) and plasma glycocalicin, a proteolytic fragment of platelet glycoprotein Ibα, in 13 patients with myelodysplastic syndrome (MDS), aplastic anaemia (AA) or idiopathic thrombocytopenic purpura (ITP). In the patients with AA or MDS, the TPO concentrations were remarkably increased, and their glycocalicin concentrations were decreased compared with the normal control individuals. In the patients with ITP, however, the TPO and glycocalicin levels were not changed as much as in the AA/MDS patients in spite of the same degree of thrombocytopenia. During immunosuppressive treatment of ITP patients, there was an inverse relationship between the level of TPO and the platelet count. Thus, measurements of TPO and glycocalicin levels are useful for the diagnosis of thrombocytopenia, and our results from ITP patients did not support the model which suggested the simple feedback regulation of TPO in thrombocytopenia.
Interleukin 1 receptor antagonist (IL1RA) in acute leukaemia: IL1RA is both secreted spontaneously by myelogenous leukaemia blasts and is a part of the acute phase reaction in patients with chemotherapy‐ induced leucopenia
Authors: Bruserud Øystein , Aasen Ingrid , Akselsen Per Espen , Bergheim Jann , Rasmussen Gro , Nesthus Ingrid
Published: July 2010

Abstract: Blast cells derived from peripheral blood of patients with acute myelogenous leukaemia (AML) were cultured in vitro and interleukin 1 receptor antagonist (IL1RA) concentrations determined in culture supernatants. AML blasts derived from patients classified as AML‐M4 and AML‐M5 subtype showed an increased release of IL1RA. IL1α and IL1β caused a similar increase in AML blast release of IL1RA, and addition of anti‐ILl antibodies decreased IL1RA release. IL1RA release from AML blasts was also increased by stem cell factor, tumour necrosis factor α (TNFα), granulocyte‐macrophage colony‐stimulating factor and macrophage colony‐stimulating factor, whereas interleukin 3, interleukin 6, leukaemia inhibitory factor and granulocyte colony‐ stimulating factor did not significantly alter IL1RA release. When investigating IL1RA serum levels, serum concentrations were decreased in acute leukaemia patients with chemotherapy‐induced cytopenia compared with healthy controls. Serum levels of both IL1RA as well as IL1β and soluble TNFα receptors increased when the leucopenic patients developed complicating bacterial infections.
Arterial and venous thrombosis in two Italian families with the factor V Arg506→ Gln mutation
Authors: Montaruli B. , Voorberg J. , Tamponi G. , Borchielli A. , Muleo G. , Pannocchia A. , van Mourik J. A. , Schinco P.
Published: July 2010

Abstract: APC resistance, due to a point mutation in factor V at amino acid position Arg506, has been identified as a major cause of inherited thrombophilia. Here we report the presence of the factor V Arg506→Gln mutation in 2 Italian families. In 1 family 3 subjects heterozygous and 2 subjects homozygous for the factor V Arg506→ Gln mutation were identified. The only subject who developed a thrombotic event was a 20‐yr‐old girl who was found to be homozygous for the factor V Arg506→Gln mutation. In the second family 10 subjects were identified to be heterozygous for the factor V Arg506 →Gln mutation; among them 2 developed a thrombotic event. In the same family 2 individuals were found to be homozygous for the mutation: the first had a myocardial infarction at age 25 yr and the second suffered from multiple episodes of deep venous thrombosis and had a stroke at age 24 yr. These data show that the risk of developing deep venous thrombosis for the carriers of the factor V Arg506→Gln mutation is high in the families investigated. Furthermore our data imply that the factor V Arg506 →Gln mutation in its homozygous form may relate to myocardial infarction and stroke.
Antibiotic proteins of polymorphonuclear leukocytes
Authors: Levy Ofer
Published: July 2010

Abstract: The polymorphonuclear leukocyte (PMN) plays an essential role in the innate defense of the mammalian host against bacterial invaders. Responding chemotactically, the PMN delivers a complex antibiotic arsenal to sites of infection. Among these cytotoxic systems is an array of antimicrobial proteins and peptides that the PMN directs at microorganisms both before (i.e. extracellularly) and after sequestration into a phagocytic vacuole. In addition to their microbicidal capacity, several of these proteins bind to and neutralize the endotoxic activity of Gram‐negative bacterial lipopolysaccharides (LPS). In this review the principle features of these antibiotic proteins are briefly summarized with emphasis on their possible actions in biological settings. In many instances, additional functions independent of cytotoxicity have been described raising the possibility that some of these proteins subserve multiple roles in inflammation.
Simplification of the blood stem cell transplantation (BSCT) procedure: large volume apheresis and uncontrolled rate cryopreservation at –80°C
Authors: Feremans W. W. , Bastin G. , Moine F. Le , Ravoet C. , Delville J. P. , Pradier O. , Wallef G. , Dupont E. , Capel P. , Lambermont M.
Published: July 2010

Abstract: Very high‐dose chemotherapy with autologous blood stem cell (BSC) rescue becomes more and more widely performed. In order to simplify the technique, a large volume apheresis programme combined with an uncontrolled rate cryopreservation at –80°C was developed. Twenty‐six patients suffering from multiple myeloma (n = 8), non‐Hodgkin's lymphoma (n = 7), dysgerminoma (n = 4), breast cancer (n = 3), Hodgkin's disease (n = 2), acute lymphoblastic leukaemia (n = 1) and acute myelocytic leukaemia (n = 1) were autografted after a classical high‐dose chemotherapy regimen. A single large volume apheresis was sufficient to obtain the threshold value of CD34+ BSC in 24/26 transplantations. The haematological recovery was favourably comparable with the previously published data obtained with controlled rate frozen BSC: median time to granulocytes > 1000/μL and to a self‐supporting platelet count > 20,000/μL, respectively, 10.5 and 12 d. The treatment‐related mortality was confined to 1/26 BSCT. These results indicate that this easy and cost‐saving policy of BSCT is efficacious and safe: sustained long‐term haematopoiesis, reduced morbidity and mortality were observed.
The effect of desmopressin on platelet aggregation defect in systemic amyloidosis: a preliminary report
Authors: Demiroǧlu Haluk , Barişta Ibrahim , Gürsoy Murat , Oymak Oktay , Diindar Semra
Published: July 2010

Abstract: Systemic amyloidosis may often be complicated with haemorrhagic tendency. The causes of this manifestation are factor deficiencies, hyperfibrinolysis and vasculopathy. In order to investigate the role of platelets, if any, we performed platelet aggregation tests with different aggregants in 10 patients with systemic amyloidosis due to familial Mediterranean fever and 10 healthy controls. Platelet aggregation was defective with different aggregants (ADP, epinephrine, collagen) in patients compared with controls. Platelet aggregation tests repeated after desmopressin (DDAVP) administration were normalized. These findings may suggest a role of a platelet aggregation defect in haemorrhagic diathesis complicating systemic amyloidosis. DDAVP may benefit patients with this disease in case of bleeding and before surgical interventions.
Flow cytometric analysis of megakaryocyte ploidy in chronic myeloproliferative disorders and reactive thrombocytosis
Authors: Jacobsson Stefan , Carneskog Jan , Ridell Börje , Wadenvik Hans , Swolin Birgitta , Kutti Jack
Published: July 2010

Abstract: Megakaryocyte (MK) ploidy patterns were analysed by flow cytometry in 29 newly diagnosed and previously untreated patients with chronic myeloproliferative disorders (MPD) and concomitant thrombocytosis, in 9 patients with reactive thrombocytosis (RT) and in 12 healthy individuals. Unfractionated bone marrow from routine aspirates was used. MKs were identified with a fluorescein labelled monoclonal antibody specific for glycoprotein IIIa (GPIIIa) and DNA was stained with propidium iodide. For the 12 healthy volunteers the mean modal ploidy number was 16 N; the 9 patients with RT displayed an identical MK ploidy pattern. The frequency of MKs with a ploidy ≥32 N was 45% among the patients with essential thrombocythaemia (ET) compared to 32% among the healthy volunteers (p < 0.001). MKs with ploidy number ≥ 64 N, comprising approximately 13% of the total number of MKs, was a characteristic finding in the patients with ET. Similar findings were present in 8 patients with polycythaemia vera (PV). In patients with PV 34% and 6% of the MKs displayed ploidies ≥32 N and ≥64 N, respectively. In contrast, a distinct shift towards lower ploidy number, with 63% of MKs ≤ 8 N, was found among the 4 patients with chronic myeloid leukaemia (CML). The present results indicate that by using flow cytometric analysis of MK ploidy distribution in patients with thrombocytosis, those with a reactive cause are likely to be discriminated from patients with myeloproliferative thrombocytosis, i.e. PV and ET on one hand and CML on the other hand. The distinction between ET and PV, however, has to be made on other grounds.
Investigation of megakaryopoiesis in myelosuppressed bone marrow using immunogold‐silver staining (IGSS)
Authors: Selig Christoph , Kreja Ludwika , Nothdurft Wilhelm
Published: July 2010

Abstract: To determine the frequencies and differential counts of megakaryocytes after cytoreductive treatment in nucleated low‐density (1.060 g/ml) bone marrow cells (BMNC) of dogs an immunogold‐silver staining (IGSS) technique with the lineage specific monoclonal antibody 2F9 was established. This antibody recognizes the glycoprotein IIb/IIIa complex expressed on the surface of canine megakaryocytes and platelets. The IGSS technique enables not only the detection of megakaryocytes occurring at a low frequency (0.1–0.2%), but also the discrimination between the different maturation stages of megakaryocytes due to cell size, nuclear morphology and cytoplasmic staining. By the use of this technique, small lymphoid megakaryocytic cells were identified. Comparable numbers of megakaryocyte colony‐forming cells in 2F9‐depleted and nondepleted BMNC suspensions (25.7 + 5.0 vs. 25.3 ± 5.1 Meg‐CFC/105 BMNC) indicate that these small 2F9 positive cells are nonclonogenic precursors of megakaryoblasts. To prove the applicability of IGSS, serial examinations of bone marrow samples from dogs treated with recombinant human interleukin‐6 (IL‐6) after exposure to 2.4 Gy total body irradiation (TBI) were performed. The results of the microscopic evaluation indicate that, in the recovery phase after TBI, IL‐6 induced an earlier and stronger increase in megakaryocyte frequency in comparison to the control. Interestingly, all maturation stages of the megakaryocytic lineage took part in this IL‐6 induced improvement of megakaryocyte recovery.
Immune recovery after autologous or rhG‐CSF primed PBSC transplantation
Authors: Rosillo M. C. , Ortuño F. , Moraleda J. M. , Lozano M. L. , Heras I. , De Arriba F. , Vicente V.
Published: July 2010

Abstract: We performed a prospective study in 17 consecutive patients following autologous bone marrow (BM) or rhG‐CSF primed peripheral blood stem cell (PBSC) transplantation, with the objective of comparing immune recovery between both procedures and to evaluate results in rhG‐CSF mobilized peripheral blood stem cell transplantation (PBSCT). Kinetics of immune reconstitution showed differences, with a faster recovery of CD3+ and CD8+ T cells, and a more rapid and sustained recovery of CD8±/CD56+ natural killer (NK) cells in the PBCSCT group. Autologous bone marrow transplantation (ABMT) was associated with a improved reconstitution of the CD19+/CD5± subpopulation. Moreover, rhG‐CSF mobilized PBSCT generated a greater recovery of CD8±/CD56+ cells than previous data concerning transplantation with peripheral blood (PB) progenitors collected after myelosuppressive chemotherapy or myelosuppressive therapy plus rhG‐CSF. Our results show differences in the rate and pattern of B and T lymphocytes reconstitution after ABMT and PBSCT. Additionally, we state an enhancement of CD56+ cells in patients undergoing PBSCT mobilized solely using rhG‐CSF.
FLAG (Fludarabine, Cytarabine, G‐CSF) as a second line therapy for acute lymphoblastic leukemia with myeloid antigen expression: in vitro and in vivo effects
Authors: Visani Giuseppe , Tosi Patrizia , Zinzani Pier Luigi , Manfroi Silvia , Ottaviani Emanuela , Cenacchi Annarita , Carrara Paola , Clavio Marino , Gobbi Marco , Tura Sante
Published: July 2010

Abstract: Thirteen consecutive adult patients with primary refractory (n = 5) or relapsed (n = 8) acute lymphoblastic leukemia (ALL) were treated by an induction schedule (FLAG) consisting of Fludarabine (30 mg/sqm/d) plus high dose Cytarabine (HD‐ara‐C: 2 g/sqm/d) (d 1–5) and G‐CSF (from d 0 to polymorphonuclear recovery). Patients achieving complete remission (CR) were administered a second FLAG course as consolidation and were then submitted to an individualized program of post‐remission therapy, depending on the patient's age and performance status. CR was achieved in 8/12 evaluable cases (67%). The median CR duration was 22.5 w. CR attainment was significantly related to the co‐expression of lymphoid and myeloid antigens. ALL/My+ patients achieved CR in 6/6 evaluable cases vs. 2/6 for ALL/My‐. In vitro 3H ara‐C incorporation into cellular DNA resulted significantly increased by Fludarabine (in 7/9 tested cases) and, furthermore, by the association of Fludarabine‐G‐CSF in 5 evaluable ALL/My+ cases; in contrast, no effect of G‐CSF addition to Fludarabine was observed in 4 ALL/My–. Myelosuppression was observed in all patients: the median time to neutrophils >0.5 × 109/l was 16.3 d (range 13–22) and 16.2 d (range 9–29) to platelets>20 × 109/l. Nonhematological toxicity was minimal. In conclusion, FLAG is an active and tolerable combination in refractory ALL, particularly in cases with myeloid antigen expression where G‐CSF appears to improve efficacy, probably increasing ara‐C incorporation into the DNA of leukemic cells.
p16INK4/p15INK4B gene inactivation is a frequent event in malignant T‐cell lines
Authors: Brandter Laura Borgonovo , Heyman Mats , Rasool Omid , Liu Yie , Grandér Dan , Einhorn Stefan
Published: July 2010

Abstract: The cell cycle regulators p16INK4 and p15INK4B have been mapped to the minimal region of overlap for chromosome 9p21 deletions, observed in a number of malignancies, suggesting that they could be tumor suppressor genes (TSGs). In the case of pl6INK4 this has been further substantiated by the finding of small intragenic mutations. In this study we have investigated the p16INK4 and p15INK4B genes in 16 malignant T‐cell lines by means of Southern blot, PCR and sequence analysis. p16INK4 allelic deletions occurred in 15 of 16 cell lines; 12 of which were homozygous and 3 hemizygous. In 1 cell line (DND 41) the remaining p16INK4 allele carried a microdeletion of 29 bp of exon 2, supporting the concept that p16INK4 is a target TSG for deletions on 9p21. Most p16INK4 deletions also included the p15INK4B gene. However, 4 of the cell lines deleted for p16INK4 showed no evidence of p15INK4B loss, indicating that p15INK4B is not the target in these cell lines.
Strong increase in the percentage of the CD8bright+CD28‐ T‐cells and delayed engraftment associated with cyclosporine‐induced autologous GVHD
Authors: Garin Laure , Rigal Dominique , Souillet Gerard , Bernaud Janine , Mérieux Yves , Philippe Noel
Published: July 2010

Abstract:  Four children with acute lymphoblastic leukaemia had autologous bone marrow (BM) or peripheral stem cell (PSC) transplantation with low dose of cyclosporine (CsA, 1 mg/kg/d i.v. during the first 28 d) to induce an autologous GVHD (auto‐GVHD). Two children did not have clinical auto‐GVHD and they relapsed 3 and 4 months after treatment. The 2 other children had clinical signs of auto‐GVHD (grade I and grade II): they both are in complete remission but after a first normal haematological recovery they had a prolonged period of aplasia until month 9 for 1 patient and still persistent at month 7 in the other case. We studied lymphocyte subsets reconstitution after transplantation in these patients. All patients had an important decrease in the CD4/CD8 ratio related both to a strong decrease in the CD4+ cells and a strong increase in the CD8+ cells. Most of the CD8+ cells were of the CD8bright+CD28‐ phenotype. These CD8bright+CD28‐ T‐cells represented from 33% to 68% of the total lymphocytes. We discuss the role of these cells after autologous transplantation with CsA, and wonder if these cells could mediate cytotoxicity. In conclusion, among 4 children who received autologous BM or PBC transplantation with low dose of CsA, we observed a complete remission after an auto‐GVHD and a prolonged period of aplasia in 2 patients and a relapse of leukaemia in 2 other patients. All these 4 patients had an increase in the CD8bright+CD28‐ T lymphocytes.
Analysis of CD34‐positive cells in bone marrow from patients with myelodysplastic syndromes and acute myeloid leukemia and in normal individuals: a comparison between FACS analysis and immunohistochemistry
Authors: Kanter‐Lewensohn Lena , Hellström‐Lindberg Eva , Kock Yvonne , Elmhorn‐Rosenborg Annika , öst Åke
Published: July 2010

Abstract:  In patients with myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML), expression of the hematopoietic stem cell marker CD34 has been associated with a poorer prognosis. CD34 is usually analyzed by flow cytometry (FC), but may also be analyzed using immunohistochemistry (IH). The present study was undertaken to compare these 2 methods. Bone marrow from 16 patients with MDS and 12 with AML and from 12 healthy young volunteers was studied. The expression of CD34 was analyzed with FC on fresh bone marrow cells and with IH on sections of paraffin‐embedded bone marrow. The correlation between FC and IH was good both for patients with MDS (p<0.0001) and AML (p<0.01). However, in patients with a high number of CD34‐positive cells, the FC method seemed to result in a higher percentage of positive cells compared to the IH method. In normal bone marrow, the ratio between the percentage of CD34‐positive cells and the percentage of bone marrow blasts was approximately 0.8. In the whole group of MDS patients, this ratio was 1:1, while in patients with refractory anemia (RA) and ring sideroblastic anemia (RAS) it was 1.6. Patients with MDS differed significantly from patients with de novo AML, who showed a ratio of only 0.23 (p<0.01). We conclude that the FC and IH methods for measuring expression of CD34 are well‐correlated in MDS and reasonably well correlated in AML. A stem cell phenotype is more commonly expressed on precursor cells from patients with MDS than from patients with AML.
Activated human platelets express β2 integrin
Authors: Philippeaux Marie Marthe , Vesin Christian , Tacchini‐Cottier Fabienne , Piguet Pierre F.
Published: July 2010

Abstract:  The expression of molecules of the β2 integrin family (CD11a, CD11b, CD11c and CD18) was explored on 2 human megakaryocytic cell lines and on platelets from different donors by immunofluorescence and flow cytometry using a large panel of mAb. CD11a, CD11b, CD11c and CD18 were detected on the megakaryocytic cell lines DAMI and HEL. A low and variable expression of CD11a, CD11b and CD18 determinants was also detected on resting platelets: this expression was markedly increased when platelets were activated by thrombin. Expression of CD18 was closely correlated to that of CD11a or CD11b when comparing the fluorescence intensity observed in different experiments. In presence of Ca++, platelets did bind to a RAJI cell line which exhibits a high expression level of CD54. This binding was increased when platelets were activated by thrombin and was decreased by an anti CD11a, CD18 and anti CD54 mAb. This study indicates that human platelets express molecules of the β2 integrin family, when activated, which allows them to bind to CD54 bearing cells.
Flow cytometric measurement of DNA S‐phase in human bone marrow cells: correcting for peripheral blood contamination
Authors: Abrahamsen Jenny Foss , Smaaland Rune , Skjærven Rolv , Laerum Ole Didrik
Published: July 2010

Abstract:  The relationship between bone marrow (BM) cells with S‐phase DNA content and the amount of peripheral blood contamination estimated as percentage lymphocytes+monocytes (L+MO) present in BM samples has been investigated in a total of 136 BM aspirates and biopsy expellates from 35 hematologically healthy individuals. A significant negative correlation was demonstrated between total, erythroid and myeloid BM cells in S‐phase and the percentage of L+MO in the aspirates (r = 0.84, 0.57 and 0.49, respectively; p<0.0001). Based on the equation of the slope of the regression line, a correction formula adjusting the measured value of BM cells in S‐phase to varying amounts of L+MO percentage has been worked out for the total and erythroid BM cells. In contrast, highly proliferating myelomonocytic cells and CD34+ cells did not show any significant correlation between cells in S‐phase and percentage L+MO, indicating that peripheral blood contamination of BM aspirates is not a problem regarding kinetic investigations of these cells. In conclusion, the described flow cytometric method of analysing BM aspirates estimates the degree of peripheral blood contamination, as well as make possible a correct estimation of the DNA synthesis of several BM populations. The method is especially applicable when frequent BM sampling is required.
Production of tumor necrosis factor and granulocyte colony stimulating factor by bone marrow accessory cells in myelodysplastic patients
Authors: Mencoboni M. , Castello G. , Lerza R. , Haupt E. , Ballarino P. , Cerruti A. , Bogliolo G. , Pannacciulli I.
Published: July 2010

Abstract:  This paper reports on the production of tumor necrosis factor (TNF) and granulocyte macrophage colony‐stimulating factor (GM‐CSF) by cultured mononuclear adherent cells derived from bone marrow of 25 patients affected by myelodysplastic syndrome (MDS) of different FAB subtypes. Mean production of GM‐CSF was much lower than in controls, without significant differences among different subtypes. Mean production of TNF was similar in MDS patients and in controls, but noteworthy differences were observed between patients with RA, RAEB and RAEB‐t and patients with RARS and CMML. Growth of bone marrow granulocyte macrophage and erythroid progenitors did not correlate with TNF and GM‐CSF production, although in MDS subtypes with higher GM‐CSF levels, colony growth was slightly higher than in subtypes with lower GM‐CSF production.
Lymphocytes in β‐thalassemia/HbE: subpopulations and mitogen responses
Authors: Wanachiwanawin W. , Phucharoen J. , Pattanapanyasat K. , Fucharoen S. , Webster H. K.
Published: July 2010

Abstract:  Lymphocyte subpopulations and proliferative responses to mitogens of 24 β‐thalassemia/HbE patients were studied and compared with those of 23 healthy controls. Results of the study were analyzed in correlation with clinical aspects i.e. severity of disease (anemia), frequency of infections and iron status. T(CD3+) lymphocytes were found to increase in thalassemic patients compared to normal controls. The CD4‐ or CD8‐positive lymphocytes and CD4/CD8 ratio were not statistically different from normals. Without mitogen, lymphocytes from thalassemic patients incorporated more [3H]Tdr than those from normal controls. Stimulation index (SI) of these cells after various mitogens were lower than in normal subjects. The observations were more obvious in patients with severe disease (severe anemia) and those who had frequent infections. These findings suggest that lymphocytes from thalassemic patients are activated in vivo. Whether these cells are less efficient in response to new or previously unexposed antigens remains to be proven.
Assessment of spleen size using gamma camera scintigraphy in newly diagnosed patients with essential thrombocythaemia and polycythaemia vera
Authors: Carneskog Jan , Wadenvik Hans , Fjälling Martha , Kutti Jack
Published: July 2010

Abstract:  By using gamma camera imaging the spleen size was assessed in 18 consecutive patients with essential thrombocythaemia (ET) and in 18 consecutive patients with polycythaemia vera (PV). All ET and PV patients were newly diagnosed and had not received any myelosuppressive therapy prior to study. The spleen areas in both posterior and left lateral projections were determined. Eighteen consecutive patients with idiopathic thrombocytopenic purpura (ITP) served as a control group since by definition they do not present with splenic enlargement; in these latter subjects the mean posterior and left lateral splenic areas were almost identical (48 ± 15 and 47 ± 17 cm2, respectively). In comparison with this control group patients with ET and PV had significantly larger spleens. In both ET and in PV patients the left lateral spleen scan area exceeded the posterior one. Patients with PV had larger splenic areas in both projections than did patients with ET, but the differences were not statistically significant. Compared to the ITP patients it was found that at least 50% of the ET patients and at least 61% of the PV patients at diagnosis presented with splenomegaly.
Influence of intravascular molecular weight of hydroxyethyl starch on platelets
Authors: Treib J. , Haass A. , Pindur G. , Treib W. , Wenzel E. , Schimrigk K.
Published: July 2010

Abstract:  Complications concerning the blood coagulation have been observed repeatedly after administration of highly substituted, high molecular weight hydroxyethyl starch (HES), but it has not been examined as to how intravascular molecular weight and degree of substitution of HES influence platelet number and volume after repeated administration. Thirty patients with cerebrovascular diseases were treated for 10 days with hemodilution. 500 to 1500 ml of HES 200/0.62 (n = 10), HES 200/0.5 (n = 10) or HES 40/0.5 (n = 10) were infused daily. During the first days, the number of platelets was not lowered beyond the dilution effect, but at the end of the therapy the number of platelets had increased in all 3 groups beyond the initial value. Platelet volume was lowered significantly in the 3 groups. HES 200/0.62 caused the largest drop in platelet volume (‐10%, p<0.01). A possible explanation could be that HES macromolecules are attached to platelets or are phagocytized by them. The larger platelets are then broken down and, to compensate the loss, more thrombocytes are released. A correlation between the molecular weight of HES and the breakdown rate of the platelets can be suspected, because HES 200/0.62 had the highest intravascular mean molecular weight (121 kD) and the largest effect on platelet volume.
Increased but highly dispersed levels of plasma glycocalicin in patients with disseminated intravascular coagulation
Authors: Kunishima Shinji , Kobayashi Sentaro , Naoe Tomoki
Published: July 2010

Abstract:  Thrombocytopenia is one of the most common laboratory manifestations of disseminated intravascular coagulation (DIC). To investigate whether thrombocytopenia in DIC is indeed due to platelet consumption, we measured the plasma levels of glycocalicin, a proteolytic fragment of the platelet membrane glycoprotein (GP) Ibα, a component of the GPIb/IX complex, in patients with solid tumors either with DIC (n = 18) or without DIC (n = 18). Patients with DIC had significantly elevated levels of glycocalicin (2.53 ± 1.59 mg/l; n = 18, p<0.05) compared to those without DIC (1.50 ± 0.41 mg/l; n = 18), indicating enhanced platelet consumption, or destruction, in patients with DIC. However, glycocalicin concentrations had a wide range (0.18–5.74 mg/l), and 3 DIC patients had levels lower than the normal lower limit. These findings suggest that, in patients with DIC, thrombocytopenia is not always due to increased platelet consumption, but it may be due to decreased production of platelets. Determination of plasma glycocalicin concentrations is an easy way to identify thrombocytopenia, due to bone marrow insufficiency in these patients.
Meeting the challenge of systemic fungal infections in cancer: nursing implications
Authors: Antrum J.
Published: July 2010

Abstract:  Several factors have contributed to the increasing incidence of fungal infections over the last 2 decades, including the emergence of Acquired Immune Deficiency Syndrome, increased use of myelotoxic chemotherapy and organ transplantations, prolonged use of broad spectrum antibiotics and aggressive intensive care procedures. The two most common opportunistic fungal infections seen today are caused by Candida spp. and Aspergillus spp. Systemic fungal infections in immunocompromised patients have an extremely high mortality and require aggressive therapy. Skill in identifying early clinical features is therefore crucial and the multifaceted role of the nurse is of major importance in the management of at‐risk patients. Nurses are the key resource in the prevention, early detection and treatment of fungal infection. All neutropenic patients should be thoroughly assessed at least twice a day, with special attention given to the most frequent sites of infection – the oral mucosa, lungs, skin, venepuncture sites and perineal area. Although fever is the hallmark of infection, it may be absent in the neutropenic patient who is unable to mount an adequate inflammatory response. It may also be masked by the use of certain drugs, such as steroids or analgesics. When systemic fungal infection is suspected, seriously ill patients require immediate antifungal therapy. Amphotericin B is currently the only agent with a sufficiently broad spectrum of activity to cover all the most common pathogens. Although conventional amphotericin B is effective, however, the required doses often carry significant toxicity, particularly nephrotoxicity. The new, lipid‐based forms of amphotericin B, such as Abelcet®, are indicated for and have been shown to be effective in patients with severe systemic and/or deep mycoses in whom conventional amphotericin B has proven ineffective or is contraindicated because of renal impairment, and in patients who have failed to respond to other antifungal agents.
The changing epidemiology of fungal infections: are the lipid‐based forms of amphotericin B an advance?
Authors: Herbrecht R.
Published: July 2010

Abstract:  The incidence of invasive fungal infections is increasing and new fungal species are emerging as important pathogens. In cancer patients, the main risk factor for the development of systemic fungal disease is severe, prolonged neutropenia. Other factors, such as mucosal damage, presence of a central venous line, immunosuppressive therapy and treatment with broad spectrum antibiotics are contributory. Candida spp. are the fungi most commonly isolated in neutropenic patients. There has been a dramatic increase in non‐C. albicans species, such as C. glabrata and C. krusei, largely as a result of extensive prophylactic and therapeutic use of fluconazole, to which these species are largely resistant. In neutropenic patients with candidaemia, amphotericin B is the drug of choice although the conventional formulation may be poorly tolerated. Lipid‐based forms of amphotericin B, such as Abelcet®, are better tolerated and can be given at a much higher dose and should therefore be considered in patients who fail on or are intolerant to the conventional agent. Aspergillosis is the second most frequent fungal infection in neutropenic patients. Primary invasive aspergillosis usually presents on chest X‐ray with lung lesions and the brain is a frequent site of secondary infection. Fluconazole is inactive against Aspergillus spp. and amphotericin B is the standard treatment. Again, lipid‐based forms are better tolerated than the conventional formulation in this setting, and have been shown to achieve response rates of 60% or more in a number of trials. Other potentially life‐threatening fungal infections in which lipid‐based amphotericin B may play an important therapeutic role in the future include cryptococcosis (increasingly problematic in AIDS patients), trichosporonosis, fusariosis and mucormycosis. Further randomized studies should be performed in a range of fungal infections to compare Abelcet®with conventional amphotericin B and other lipid‐based antifungal agents.
Amphotericin B Lipid Complex (Abelcet®) in the treatment of invasive mycoses: the North American experience
Authors: Lister J.
Published: July 2010

Abstract:  Abelcet®, or Amphotericin B Lipid Complex, is a unique formulation, comprising an equimolar mixture of amphotericin B complexed with two lipids. In preclinical studies, Abelcet® was clearly demonstrated to be less toxic than amphotericin B desoxycholate and to be effective in models where amphotericin B was ineffective at its maximum tolerated dose. Pharmacokinetic studies in animals also showed that the concentration of Abelcet® in blood is similar or reduced compared to levels seen with conventional amphotericin B, with accumulation in the liver, lungs and spleen. Phase I clinical trials determined the optimum tolerated dose of Abelcet® to be 5 mg/kg d‐1. Data are now available for 228 cases (including 51 paediatric cases) of invasive fungal infection treated with Abelcet® in an open‐label emergency‐release protocol. All patients had to have failed on previous amphotericin B or other conventional antifungals, or to have unacceptable toxicity on amphotericin B, or underlying renal disease, or nephrotoxicity due to other drugs. Abelcet® was administered at a dose of 5 mg/kg d‐1 for 4 wk. Approximately one‐third of patients had candidiasis, one‐third aspergillosis and one‐third other infections, including fusariosis. Of 183 cases evaluable for response, 126 (69%) had a clinical response (cure or improvement) which was mycologically confirmed in 55% (61/110 tested). Results in paediatric cases were similar to or better than those seen in the group as a whole. When comparisons were made between cases with different types of infection, underlying disease/immunosuppressive disorder, and degree of neutropenia, the response rates were very consistent from group to group. Treatment with Abelcet® was well tolerated and mean serum creatinine levels actually declined during therapy, particularly in patients with pre‐existing renal dysfunction.
Developmental change of megakaryocyte maturation and DNA ploidy in human fetus
Authors: Ma D. C. , Sun Y. H. , Chang K. Z. , Zuo W.
Published: July 2010

Abstract:  Megakaryocytes in fetal livers obtained from 30 water‐balloon aborted normal fetuses of 3 to 6 months' gestation, in the bone marrow from the same 30 fetuses, and another 9 fetuses of 7 to 8 months' gestation and in the normal bone marrow of adults were analyzed by immunocytochemical staining for size and maturation stage distribution and by flow cytometry for ploidy distribution simultaneously. In human fetuses, megakaryocytes showed a shift during ontogenesis from smaller towards larger size and from less mature towards a more mature stage with advancement of gestation. This was accompanied by a significant progressive shift to higher ploidy. However, the proportions (78.64%) of hypoploidy (≤8N) megakaryocytes in bone marrow of 7–8 months' gestation fetuses was still much higher than that (33.32%) in human adults (p<0.05), with the proportion of hyperploidy (≤16N) megakaryocytes lower than that (67.86%) in human adults. This result indicated that megakaryocyte polyploidization may be retarded or inhibited during development.
Effects of combinations of stem cell factor and hemin on erythropoiesis after azidothymidine treatment of immunosuppressed mice
Authors: Hamburger Anne W.
Published: July 2010

Abstract:  Recombinant cytokines such as stem cell factor (SCF) are currently being tested for the ability to ameliorate 3′azido‐3′deoxythymidine (AZT) induced anemia in AIDS patients. Recently, we demonstrated that SCF and hemin in vitro greatly increased the resistance of burst‐forming units erythroid (BFU‐E) to AZT. We therefore attempted to ameliorate AZT‐induced anemia in vivo using SCF and hemin in immunodeficient LP‐BM5 infected (MAIDS) mice. SCF and hemin were administered with oral AZT for 3 wk and the effects on erythropoiesis examined. Hemin significantly increased hematocrits of AZT‐treated mice and control mice. However, SCF and SCF–hemin combinations failed to raise hematocrits. Reticulocyte numbers were significantly consistently increased only in hemin‐treated mice receiving AZT. The numbers of CFU‐E were increased in bone marrow of AZT‐treated mice receiving hemin. Therefore, SCF did not enhance the erythropoietic effect of hemin in AZT‐treated immunodeficient mice.
Maintenance therapy with interferon‐α (IFN‐α) versus IFN‐α plus chemotherapy in multiple myeloma (MM)
Authors: Zervas K. , Pouli A. , Perifanis V. , Papanastasiou K. , Chatziyianni M. , Mitsouli C. , Maniatis A. ,   Greek Myeloma Study Group
Published: July 2010

Abstract:  Results of studies using IFN‐α treatment for maintaining remission and prolonging survival in multiple myeloma (MM) are in conflict and trials seeking optimum use for this biological response modifier are continuing. Between 1989 and 1993 a prospective randomized multicentre trial was undertaken to evaluate the role of the combination of IFN‐α with chemotherapy (CT) in maintenance treatment of MM. For remission induction, in patients 65 yr or younger, we used VAD (group A) and for the remaining Melphalan and Prednisone (MP) (group B). For maintenance, patients were randomized to receive IFN‐α 3times106 i.u. s.c. t.i.w. (group I) or alternating monthly cycles of IFN‐α and CT. The CT cycles were also alternated (VAD, MP, CP) in an effort to prevent the development of multidrug resistance. Median survival of the two maintenance groups from randomization (36 months for group I and 31 months for group II, p=0.3) as well as response duration (13 months in group I and 15 months in group II, p=0.95) were similar. Toxicities were more pronounced both with VAD induction and in the combination maintenance arm. The addition of chemotherapy to the IFN maintenance regimen in MM did not have an advantage over IFN alone.
Serum procollagen III peptide concentration in iron overload
Authors: Jensen P. D. , Heickendorff L. , Helweg‐Larsen H. M. , Jensen F. T. , Christensen T. , Ellegaard J.
Published: July 2010

Abstract:  Patients with severe iron overload may develop hepatic fibrosis due to iron toxicity. Unfortunately, the follow‐up of the fibrogenic activity during treatment by histological examination of tissue biopsies carries potential side effects, and may therefore not be justified ethically. Recently, the serum concentration of procollagen type III peptide (S‐PIIINP) has been shown to be a valid serum marker of the activity of collagen metabolism in conditions with hepatic fibrosis unrelated to iron overload. In order to evaluate the potential usefulness of this test in patients with fibrosis due to iron overload, we investigated the relationship between the PIIINP serum concentration and the size of iron overload in 18 patients with hereditary haemochromatosis (HH) and in 14 patients with transfusional iron overload. A close correlation was found between S‐ferritin and S‐PIIINP (r=0.73, p<0.0001). Follow‐up of 6 patients during iron depletion treatment revealed a normalization of the serum aminotransferase concentration before normalization of S‐PIIINP was found. This may indicate that excess iron directly induces an increase in fibrogenesis rather than the increased fibrogenesis is secondary to hepatocellular injury caused by iron excess. Thus, serial measurements S‐PIIINP may be useful in follow‐up of the fibrogenic process due to iron overload.
A long‐term follow‐up study of interferon treatment for chronic hepatitis C in Japanese patients with congenital bleeding disorders
Authors: Yamada M. , Fukuda Y. , Koyama Y. , Nakano I. , Urano F. , Isobe K. , Takamatsu J. , Imoto M. , Hayakawa T.
Published: July 2010

Abstract:  Twenty‐one HIV negative Japanese patients with chronic hepatitis C who had congenital bleeding disorders, 15 hemophilia A, 3 hemophilia B, 1 von Willebrand's disease, 1 afibrinogenemia and 1 thrombasthenia, were treated with 9 million units 3 times a week of natural interferon (IFN)‐α for 6 months. They were followed, biochemically and virologically, for at least 18 months after therapy discontinuation to evaluate the long‐term results. Liver biopsy, hepatitis C virus (HCV) genotyping and quantification of viral load by polymerase chain reaction (PCR) were performed to identify the predictors of a favorable response to IFN treatment. One male patient with hemophilia A dropped out because of general fatigue and was excluded from evaluation. Ten (50.0%) patients continued to be HCV RNA negative in serum together with normal ALT levels throughout the study. Subtype 1b and a high level of viremia significantly associated with an unfavorable outcome on the response to IFN although liver histology was not definitive for predicting the response. We concluded that a 6‐month treatment with high doses of natural IFN‐α was effective in inducing a long‐term response without relapse of viremia in 50% of chronic hepatitis C patients with congenital bleeding disorders and that HCV subtype and pretreatment level of viremia were useful predictors of the response to IFN in treating such patients.
Effect of interferon‐α on immunoglobulin production by peripheral blood mononuclear cells in multiple myeloma
Authors: Säily M. , Koistinen P. , Laine S. , Soppi E. , Savolainen E‐R.
Published: July 2010

Abstract:  To test a hypothesis that interferon‐α (IFN) treatment might restore normal immunoglobulin (Ig) production in multiple myeloma (MM), the effect of IFN on Ig isotype (IgG and IgA) production by peripheral blood (PB) and bone marrow (BM) mononuclear cells (MNCs) in MM patients was analyzed by ELISA. IFN at a concentration of 1000 U/ml was found to enhance IgA production by PB MNCs in IgA‐MM and had a trend to stimulate IgG production in IgG‐MM. The effect of IFN on nonparaprotein Ig isotype production was more variable, with mostly neutral or inhibitory effects being seen in both the MM subtypes. In contrast to the influences observed in MM patients, IFN at the same concentration inhibited both IgG and IgA production by PB MNCs in healthy controls. In studying BM cells, IFN was found to reduce IgA production in IgA‐MM, but had a neutral effect on IgG production in IgG‐MM. In the controls, the production of both the IgG and the IgA isotypes by BM MNCs was decreased by IFN.

On the basis of these results it seems that the disease itself somehow affects the Ig‐producing cells in MM, when measured as different responses of the cells to exogenous IFN in vitro. The results do not support the hypothesis that IFN treatment could restore normal Ig production in MM patients.
Decreased IL‐1β and TNFα secretion in long‐term bone marrow culture supernatant from Fanconi's anaemia patients
Authors: de Cremoux P. , Gluckman E. , Podgorniak M.P. , Menier C. , Thierry D. , Calvo F. , Socié G.
Published: July 2010

Abstracts:  Recent studies have shown that abnormalities of cytokine and lymphokine secretion are involved in the pathophysiology of Fanconi's anaemia (FA). In the present study, we quantified IL‐1β, IL‐1 receptor antagonist (IL‐1Ra), IL‐6 and TNFα protein levels in the supernatant of long‐term cultures generated from BM cells of FA patients. Cell‐free conditioned medium from long‐term bone marrow culture was harvested every week at confluence and tested for interleukin secretion. IL‐1β, IL‐1Ra, TNFα and IL‐6 protein secretion was assessed using immunoassays. IL‐6 secretion was similar between controls and FA supernatants from wk 1 to wk 4. TNFα released from FA cells was consistently found at very low levels compared to control cells during the first 3 wk. Furthermore, secretion of IL‐1β by cells from FA was always more than 2 standard deviations below the value of IL‐1β found in normal donor cells from wk 1 to wk 4. In conclusion, in addition to a stem cell defect, a marked decrease in IL‐1β and TNFα secretion may be one of the mechanisms leading to bone marrow failure in individuals with Fanconi's anaemia.
Genotyping of the platelet‐specific alloantigen HPA‐5 (Bra/Brb) using polymerase chain reaction with sequence‐specific primers (PCR‐SSP)
Authors: Hou M. , Rosengren‐Kogan L. , Forsberg B. , Elmgren A. , Rydberg L. , Kutti J. , Wadenvik H.
Published: July 2010

Abstract:  A DNA‐based one‐stage technique, polymerase chain reaction with sequence‐specific primers (PCR‐SSP) was developed for genotyping of the platelet specific alloantigen HPA‐5 (Bra/Brb). Sequence‐specific primers, matching the wild type and the point mutation responsible for the HPA‐5 (Bra/Brb) phenotype, were constructed. Conjointly a fragment of the gene coding for glycoprotein (GP) IIIa was amplified as an internal control of the enzyme reaction. Using these HPA‐5 (Bra/Brb) sequence‐specific primers the correct fragment of the GPIa gene was amplified, as evidenced by the PCR product size, the restriction map and by the nucleotide sequence. This assay was applied on 187 Swedish blood donors; 157 individuals were found to have a homozygous HPA‐5a (Brb/Brb) genotype and 30 individuals a heterozygous HPA‐5a,b (Bra/Brb) genotype. None of the donors was found to display a homozygous HPA‐5b (Bra/Bra) genotype. Thus, the (HPA‐5b) Bra antigen frequency in this population will be approximately 16.0% with a gene frequency of 8.0%. It is concluded that this assay is an attractive technique for genotyping of the HPA‐5 (Bra/Brb) alloantigens on genomic DNA. The technique can replace serological alloantigen typing, especially in cases where platelets and rare human alloantisera are not available.