bluuebunny/crossref_metadata_2025 huggingface dataset:
Hb Trollhättan [β20(B2)Val& RR; Glu] — a new haemoglobin variant with increased oxygen affinity causing erythrocytosis
Authors: Landin Britta , Berglund Stig , Lindoff Bertil
Published: July 2010

Abstract: Erythrocytosis is sometimes caused by the presence of haemoglobin (Hb) variants with increased oxygen affinity. Here we describe a β‐globin variant found by isoelectric focusing (IEF) of Hb from a 23‐year‐old Swedish male with moderate erythrocytosis. Amplification of DNA corresponding to the β‐globin gene and subsequent nucleotide sequencing revealed heterozygosity for a GTG& RR; GAG mutation at codon 20, thus suggesting a Val& RR; Glu substitution, which was confirmed at the protein level. This mutation occurs at the same position as Hb Olympia [β20(B2)Val& RR; Met], another variant associated with erythrocytosis due to increased oxygen affinity. The novel variant, Hb Trollhättan [β20(B2)Val& RR; Glu], which was also associated with increased oxygen affinity, was shown to be present in three generations of the patient's family.
Peripheral blood mononuclear cells from patients with rheumatoid arthritis suppress erythropoiesis in vitro via the production of tumor necrosis factor alpha
Authors: Katevas Prokopis , Andonopoulos Andrew P. , Kourakli‐Symeonidis Alexandra , Manopoulou Efi , Lafi Theoni , Makri Maria , Zoumbos Nicholas C.
Published: July 2010

Abstract: Anemia is a frequent manifestation of rheumatoid arthritis, with a probably multifactorial etiology. We investigated the effect of peripheral blood mononuclear cell supernatants (PBMCS) from rheumatoid arthritis (RA) patients on hematopoietic colony formation in vitro, by using a methylcellulose assay. PBMCS from patients suppress in vitro erythroid (BFU‐E), mixed‐lineage (CFU‐GEMM) and to a lesser degree granulocyte‐macrophage (CFU‐GM) progenitors. PBMCS from anemic RA patients were more suppressive for BFU‐E than those from non‐anemic patients. Addition of antibodies to tumor‐necrosis factor alpha (TNFα) almost completely reversed the inhibition of BFU‐E and CFU‐GEMM, but had little effect on the CFU‐GM colony formation. Antibodies to interferon‐gamma (IFNγ) and interleukin‐1 (IL‐1) were not effective. The above data suggest that PBMCS from RA patients suppress in vitro erythropoiesis via the production of TNFα; a pathogenetic role for TNFα in the anemia of RA can be implied.
TNF and IL‐6 are potent growth factors for OH‐2, a novel human myeloma cell line
Authors: Börset Magne , Waage Anders , Brekke Ole Lars , Helseth Eirik
Published: July 2010

Abstract: A novel human myeloma cell line, OH‐2, was established from pleural fluid of a myeloma patient in end stage of the disease. Effects of cytokines on proliferation were analyzed by measuring uptake of 3H‐thymidine. Cell surface antigens were detected by flow cytometry. The cell line is dependent on IL‐6 for growth and proliferates in response to TNF. There is synergy between the stimulatory response of TNF and IL‐6. The cells express both the p55 and p75 TNF receptors. Neutralizing anti‐IL‐6 did not inhibit TNF‐mediated proliferation, showing that TNF acts through a pathway that is independent of IL‐6. TNF was more potent than IL‐6 in stimulating the growth of primary myeloma cultures (>99% pure) from the same patient (OH‐2‐PC), indicating that TNF in selected myeloma patients has a growth‐promoting effect equal to IL‐6. OH‐2 cells produce and secrete monoclonal IgG‐kappa.
Effects of soluble interleukin‐1 receptor and tumor‐necrosis factor receptor, respectively, on the IL‐1‐ and TNF‐α‐induced DNA synthesis of acute myeloblastic leukemia blasts in vitro
Authors: Carter Anna , Haddad Nuhad , Draxler Ilana , Tatarsky Ilana
Published: July 2010

Abstract: This study demonstrates that soluble interleukin‐1 receptor and tumor necrosis factor receptor modulate their corresponding cytokine‐induced DNA synthesis of acute myeloblastic leukemia (AML) blasts in a dose‐dependent, bimodal fashion; at lower concentrations they enhanced, while at high concentrations they inhibited, the cytokine‐mediated effects. Furthermore, the concentrations of endogenously produced IL‐1β and TNF‐a were found to be significantly (p<0.01) higher in supernatants of AML cells cultured in the presence of corresponding soluble receptors compared to their levels in supernatants of cells growing in the absence of these molecules. Our data might suggest that the attenuation of the spontaneous decay of IL‐1β as well as TNF‐a activities by soluble receptors may account for their ability to augment some of their effects.
Inherited giant platelet disorders
Authors: Jantunen Esa
Published: July 2010

Giant platelet disorders (GPD) refer to rare, usually inherited states characterized by abnormally large platelets, thrombocytopenia and bleeding tendency of variable severity. This review summarizes major clinical and laboratory features of three GPDs (Bernard‐Soulier syndrome, May‐Hegglin anomaly and gray platelet syndrome). Differential diagnosis between immunological thrombocytopenia and GPDs is important. Although rare, giant platelet disorders should be borne in mind, since bleeding tendency in some individuals may be severe and knowledge of bleeding diathesis is of importance before delivery or surgical procedures also in less symptomatic individuals.
In vitroimprovement of bone marrow‐derived hematopoietic colony formation in HIV‐positive patients by alpha‐D‐tocopherol and erythropoietin
Authors: Geissler R. Georg , Ganser Arnold , Ottmann Oliver G. , Gute Peter , Morawetz Anja , Guba Petra , Helm Eilke B. , Hoelzer Dieter
Published: July 2010

The majority of patients with progressive HIV infection develop a severe hematopoietic failure which is aggravated by the hematotoxic effect of azidothymidine (AZT) treatment. Since it was shown in a mouse model that α‐D‐tocopherol (vitamin E derivative) antagonizes the inhibitory influence of AZT on the growth of burst‐forming units‐erythrocyte (BFU‐E), it was the aim of this study to investigate whether α‐D‐tocopherol and high dosages of erythropoietin (EPO) increase the hematopoietic colony‐forming capacity of bone marrow cells from patients with progressive HIV disease and especially if they reverse the inhibitory effects of AZT. The data demonstrate that tocopherol (1–100ümol/l) significantly increases the growth of BFU‐E and colony‐forming units granulocyte‐monocyte (CFU‐GM) from HIV‐infected patients. This stimulatory effect is dose‐dependent (maximum at 30–100 μmol/l) and only occurs when the agent is present from the beginning of the cultures. EPO (5–10 U/ml) also augments the numbers of BFU‐E from HIV‐infected patients. Tocopherol equally ameliorates the growth of BFU‐E and CFU‐GM from the HIV‐positive cohort in the presence of AZT (10–100 μmol/l). For healthy controls, no such increase was observed, either with tocopherol or with higher dosages of EPO. In conclusion, both tocopherol and EPO partially reverse the myelosuppressive action of AZT in HIV‐positive patients.
Renal function in newly diagnosed multiple myeloma — A demographic study of 1353 patients
Authors: Knudsen Lene Meldgaard , Hippe Erik , Hjorth Martin , Holmberg Erik , Westin Jan
Published: July 2010

This study describes the occurrence of renal failure among 1353 newly diagnosed cases of multiple myeloma. Renal function was evaluated by serum creatinine concentration in 1353 cases, 31% of whom had renal failure at the time of diagnosis. In 1206 cases an estimation of creatinine clearance was made. When renal failure was defined by using creatinine clearance estimation, 49% had renal failure at the time of diagnosis. Renal failure was present in 24% of patients with an M component of IgG‐, 31% of IgA‐ and 100% of IgD‐type. 52% of patients with light chain disease had renal failure. The frequency of renal failure was similar in lambda‐and kappa‐light chain disease. Patients with a high excretion of Bence Jones protein in the urine (> 10 g/24 h) had renal failure significantly more often than patients with lower excretion. Renal failure was related to advanced disease; 41% of patients with stage III (Durie‐Salmon) disease had renal failure. Renal failure was found in 45% of patients with hypercalcaemia. When estimated creatinine clearance was used as a predictor of renal function, the same trends were found as mentioned above. In addition, the proportion of patients with renal failure was found to increase with advancing age.
Recombinant α2a interferon and polycythemia vera: Clinical results and biological evaluation by means of Fourier‐transform infrared microspectroscopy
Authors: Papineschi F. , Bucalossi A. , Capochiani E. , Benedetti E. , Bramanti E. , Dastoli G. , Dispensa E. , Benedetti E. , Spremolla G.
Published: July 2010

Polycythemia vera (PV) is a chronic myeloproliferative disease. The use of recombinant α2a Interferon (IFN) therapy in this disease is a novel approach. We applied Fourier‐transform infrared microspectroscopy (FT‐IR‐M) to investigate the behavior and therapeutic responsiveness of PV patients treated with IFN. A spectroscopic parameter (A1/A2) was used, corresponding to the ratio of the integrated areas of the bands at 1080 cm‐1 and at 1540 cm‐1 due to nucleic acids and proteic components, respectively, calculated on the spectra of single megakaryocytes (MKs). In previous studies, we have pointed out that MKs in PV have a surprisingly strong myeloproliferative impulse when compared to MKs from other chronic myeloproliferative diseases. Nine patients out of the 11 studied exhibited a satisfactory responsiveness to the IFN treatment. Ten patients were evaluated by the A1/A2 parameter. In 8 of these, a good agreement was seen between this parameter and the laboratory data commonly used for the assessment of this disease. The infrared parameter, which we propose, proves to be an original, reliable method for the evaluation of recombinant α2a IFN responsiveness in this disease.
Risk of kidney cancer and other second solid malignancies in patients with chronic lymphocytic leukemia
Authors: Mellemgaard Anders , Geisler Christian H. , Storm Hans H.
Published: July 2010

Patients with chronic lymphocytic leukemia (CLL) are known to to have an increased incidence of secondary cancers. We investigated the occurrence of secondary cancers in 7391 patients with CLL diagnosed between 1955 and 1988. The observed number of cancer cases was compared with the expected number of cancers calculated from national cancer incidence rates. The overall risk of cancer was significantly increased among persons with CLL. The standardized incidence ratios (ratio between the observed and the expected numbers) were 2.0 for men and 1.2 for women. Increased risks were found for cancer of the lung and prostate in men (RR = 2.0 and 1.5 respectively), renal parenchyma in both sexes (RR = 2.8 for men, RR = 3.6 for women) non‐melanoma skin cancer in both sexes (RR = 4.7 for men, RR = 2.4 for women) and sarcomas (RR = 3.3 for men, RR = 2.8 for women). Although an increased risk of cancer is to be expected solely because individuals with CLL are being physically examined frequently, it appears that the risk is significantly increased for a number of cancer sites in persons with CLL.
Heterogeneous X‐ray survival characteristics of lymphocytes in prolymphocytic leukaemia: Mathematical analysis distinguishing delayed cell death and true radioresistance
Authors: Thomson Alasdair E. R. , O'Connor Thomas W. E. , Peel William E. , Slater Nicolas G. P.
Published: July 2010

The survival of non‐dividing (G0) leukaemic lymphocytes in culture is generally too short for their radiosensitivity to be quantitatively assessed, since lethally X‐irradiated cells may show a long delay before manifestations of cell death (“interphase death”) are seen. Counts of surviving cells will therefore include both lethally‐hit cells (apparent survivors), and real survivors which have not been lethally hit. Death rates of irradiated leukaemic and normal cells show great variation between individuals, so that comparisons of radiosensitivity between different cell populations based on surviving cell counts at a single time‐point are invalid. In this study the supposed radioresistance of prolymphocytic leukaemia lymphocytes was examined in 6 patients with B‐cell disease. Survival curves were plotted from serial observations made over several days after graded X‐irradiation (0–1000 cGy). We attempted to interpret these radiation responses in terms of their dose dependence (intrinsic radiosensitivity) and time dependence (cell death rate) characteristics using the best‐fitting of four mathematical models, all based on classical “single‐hit” target theory. The apparent radioresistance shown in 4 cases could be explained by very slow death rates (T1/2 values 55–205 h) of cells proving otherwise radiosensitive (D37 values 38–123 cGy). Genuine radioresistance was found in only 1 case (actual D37 value above 2000 cGy). By ignoring delayed cell death in clinical assessments, pathological lymphocytes could be mistakenly categorised as resistant to elimination by radiotherapy.
Specific antiplatelet autoantibodies in patients with antiphospholipid antibodies and thrombocytopenia
Authors: Fabris Fabrizio , Steffan Agostino , Cordiano Immacolata , Borzini Pietro , Luzzatto Guido , Randi Maria Luigia , Girolami Antonio
Published: July 2010

By means of immunoblotting and monoclonal antibody immobilization of platelet antigens (MAIPA) we have studied the specificity of antiplatelet antibodies in patients with antiphospholipid antibodies and thrombocytopenia defined as presence of anticardiolipin IgG and a platelet count below 100 × 109/l. The study group consisted of 10 patients with systemic lupus erythematosus (SLE), 8 patients with primary anti‐phospholipid syndrome (PAPS) and 16 patients with idiopathic thrombocytopenic purpura (ITP). The comparison group was formed by 17 patients with classical chronic ITP without anticardiolipin IgG. We identified the 80–100, 130–150 and 150–170 KD surface proteins that comigrate with GPIIIa, GPIIb and GPIb and a 50–70 KD cytoplasm band by immunoblot. In patients with classical chronic ITP, the prevalence of the antiplatelet antibodies against GPIIIa was 53% on immunoblot assay and 47% on MAIPA. In ITP patients who had also anti‐phospholipid antibodies in serum, the percentage of reactivity to GPIIIa declined to 37% on immunoblot and 21 % on MAIPA but it was not statistically different from the percentage observed in patients with classical ITP. Autoantibodies to platelet surface glycoproteins were almost absent in SLE and PAPS patients, who showed a significant prevalence (78%) of IgG reactivity to the 50–70 KD internal platelet protein which was frequently encountered also in patients with ITP and aPL (56%). Our study provides additional evidence that platelet antigens in patients with phospholipid‐associated secondary immune thrombocytopenia are different from those of primary ITP, and that surface glycoproteins were not involved.
Advantage of induction therapy with all trans retinoic acid in acute promyelocytic leukaemia in a country with limited transfusion resources: A Malaysian experience
Authors: Leong K. W. , Bosco J. J. , Teh A.
Published: July 2010

Induction of remission of acute promyelocytic leukaemia (APL) needs intensive blood support (16) to prevent bleeding attributed to disseminated intravascular coagulation. Between 1989 and 1991, at the University Hospital in Kuala Lumpur, Malaysia, the remission rate of APL was only 27% with conventional chemotherapy as a result of inadequate transfusion resources. The use of all trans retinoic acid in induction therapy followed by consolidation and maintenance chemotherapy has improved the situation dramatically. Twelve patients entered the study. Ten patients achieved remission (83%), indicating how ATRA had significantly improved the results (p = 0.003). Blood component transfusions were also significantly reduced (p = 0.003). Two ethnic Chinese patients developed pulmonary leucostasis. Published Chinese (2, 6) and Japanese (11) studies have not reported this serious adverse effect. We can now state that leucostasis is not a phenomenon limited to the Western population. ATRA has proved to be extremely beneficial for patients at this centre. Early analysis also suggests that consolidation and maintenance chemotherapy has prolonged remission duration. ATRA should be made available for the treatment of APL in all countries where there are inadequate transfusion services.
Bernard‐Soulier syndrome: quantitative characterization of megakaryocytes and platelets by flow cytometric and platelet kinetic measurements
Authors: Aaron Tomer M.D  , Scharf Rüdiger E. , McMillan Robert , Ruggeri Zaverio M. , Harkers Laurence A.
Published: July 2010

Abstract: Platelets and megakaryocytes have been characterized in a Bernard‐Soulier syndrome (BSS) kindred with respect to glycoprotein (GP) membrane receptors and measurements of thrombocytopoiesis. The index patient exhibited lifelong bleeding tendency, moderate thrombocytopenia (35 × 109/1), giant platelets (mean platelet volume 12.5 μm3 compared to 7.5 ± 1.5 μm3 in normals), absent ristocetin‐induced platelet agglutination and absent binding of von Willebrand factor (vWF). Flow‐cytometric analysis revealed absent platelet binding (0–2%) of monoclonal antibodies (mAb, LJ‐P3, LJ‐Ibl and LJ‐Ibl0) directed against distinct epitopes on membrane GPIbα of the GPIb‐IX complex, and normal binding of LJ‐P4 mAb directed against GPIIb/IIIa complex (relative to increased platelet surface area). Marrow megakaryocytes also failed to express GPIb‐IX complex, but demonstrated normal expression of GPIIb/IIIa. Among 6 asymptomatic family members, the patient's mother and 2 of his 4 children exhibited approximately 50% binding of anti‐GPIbα mAb to their platelets by both flow cytometry and direct binding studies using 125I‐vWF, 125I‐LJ‐Ibl and 125I‐LJ‐Ibl0 mAb. Marrow megakaryocytes were increased in the average cell volume and cytoplasmic granularity with a corresponding increase in ploidy (46% > 16N compared to 22 ± 5% in normal individuals), a pattern typical of megakaryocytes stimulated by thrombocytopenia. Autologous 111In‐platelet life span was shortened to 4.1 days (compared with 9.5 ± 0.5 days in normal subjects), and the turnover of platelet mass in the circulation was near normal. The data directly demonstrate that the platelet membrane GPIb‐IX defect in BSS originates in megakaryocytes at all levels of cell maturation, and exclude the possibility that the receptor abnormality is acquired during cell maturation or after platelets are released into the circulation. Since marrow megakaryocytes exhibited cellular changes consistent with stimulated megakaryocytopoiesis, these results also suggest that thrombocytopenia in this kindred of BSS is a consequence of both decreased platelet survival and ineffective platelet production.
Predominance of myeloid antigens in CD34‐positive peripheral blood stem cells over those in bone marrow after administration of granulocyte colony‐stimulating factor
Authors: Fukuda Takahiro , Okamura Seiichi , Shimoda Kazuya , Takamatsu Yasushi , Inaba Shouichi , Harada Mine , Niho Yoshiyuki
Published: July 2010

Abstract: We analyzed the surface phenotype of CD34‐positive (CD34+) cells in peripheral blood (PB) during the period of hematopoietic recovery following myelosuppressive chemotherapy. A significantly higher proportion of PB CD34+ cells coexpressed the CD13 and CD33 myeloid antigens (80.8%, 78.1%, respectively) than did BM CD34*** cells (45.8%, 37.8%, respectively) (both p < 0.01). In particular, the CD13 positivity of PB CD34+ cells harvested with granulocyte colony‐stimulating factor (G‐CSF) was significantly higher than that of those without G‐CSF. Most of the PB CD34+ cells possessed the HLA‐DR antigen, and less than 10% of the CD34+ cells coexpressed a mature cell antigen, such as CD14, GPA or Plt‐1. The administration of G‐CSF enhanced the appearance of significantly larger amounts of CD13+ 34+ and CD33+ 34+ cells (both p<0.01). This G‐CSF mobilization also resulted in an increased number of CD 13‐34+ and CD33‐34+ cells and all types of colony‐forming cells. On the other hand, macrophage colony‐stimulating factor administration exerted little influence on the mobilization of PB CD34+ cells. Thus, G‐CSF seemed to induce not only an expansion of circulating hematopoietic stem cells but also the myeloid differentiation of stem cells.
Role of protein phosphorylation in EPO‐mediated early signal transduction: Analysis in the EPO‐reactive cell line ELM‐I‐1 transfected with a c‐fos‐enhancer/promoter‐luciferase reporter gene
Authors: Tsuda Hiroyuki , Huang Ren‐Wei , Takatsuki Kiyoshi
Published: July 2010

Abstract: To investigate the role of protein phosphorylation in the early phase of EPO‐mediated signal transduction, we EPO‐stimulated a murine erythroid cell line ELM‐I‐1 transformed by plasmids comprised of the c‐fos enhancer/promoter linked to the luciferase gene. Using this reporter gene system, we previously showed that EPO‐induced activation of the c‐fos promoter can be detected rapidly and sensitively as an elevation of cellular luciferase activity. In this study, we first examined the role of protein tyrosine phosphorylation. The tyrosine phosphatase inhibitor orthovanadate not only induced luciferase activity by itself but enhanced the action of EPO. On the other hand, the tyrosine kinase inhibitors erbstatin and herbimycin suppressed the effect of EPO. Next, the role of protein kinase C (PKC) in the EPO response was assessed. The PKC activator phorbol myristate acetate (PMA) not only induced luciferase activity by itself but enhanced the action of Epo. On the other hand, the PKC inhibitor 1‐(5‐isoquinolynyl‐sulfonyl)‐2‐methylpiperazine (H7) suppressed the effect of Epo and PMA, whereas a nonspecific protein kinase inhibitor, N‐(2‐Guanidinoethyl)‐5‐Isoquinolinesulfornamine (HA1004) inhibited the action of neither Epo nor PMA. Another known PKC inhibitor staurosporine (STSP) did not inhibit but rather enhanced the effect of Epo. This action of STSP was blocked by H7 but not by HA1004. These results suggest that the EPO‐mediated early signal transduction pathway leading to c‐fos expression involves protein‐tyrosine phosphorylation, is modulated by tyrosine phosphatase activity and is positively regulated by PKC.
Vincristine sulfate for the treatment of thrombotic thrombocytopenic purpura refractory to plasma‐exchange
Authors: Bobbio‐Pallavicini E. , Porta C. , Centurioni R. , Gugliotta L. , Vianelli N. , Tacconi F. , Billio A. , Ascari E.
Published: July 2010

Abstract: Among all the patients treated by the Italian Cooperative Group for TTP, we retrospectively reviewed the results obtained using vincristine (VCR) in 8 TTP patients (4 men and 4 women, average age: 39.25 years, range: 23–48) who did not respond to combined apheretic and pharmacologic treatment. All patients, after failing to respond to treatment, were started on VCR at the dose of 2 mg, i.v., once a week. Despite this treatment, 4 patients (50%) died 1, 7, 12 and 25 days after the first VCR dose, respectively. The other 4 patients who received VCR achieved complete remission 24, 30, 40 and 50 days from the beginning of the treatment. Total doses of VCR ranged from 2 to 6 mg in the deceased group, and from 6 to 14 mg in the cured patients. In our experience, VCR is a promising agent to treat TTP patients resistant to conventional plasma‐exchange and pharmacologic therapy.
Elevated serum homocysteine as a predictor for vitamin B12 or folate deficiency
Authors: Curtis David , Sparrow Rosemary , Brennan Leanne , Van Der Weyden Martin B.
Published: July 2010

Abstract: Tissue deficiency of vitamin B12 and folate results in an increase in serum homocysteine (sHcy). We have measured sHcy in patients with reduced serum vitamin B12 and/or red cell folate (RCF) to determine its usefulness as a discriminant for the diagnostic interpretation of reduced vitamin levels. Of 3846 patients who had serum vitamin B12 and RCF assayed, 335 (9%) had reduced vitamin levels. Multivariate analysis showed a significant association between sHcy and serum creatinine (p = 0.0001), positive intrinsic factor (IF) antibody or neutrophil hypersegmentation (NHS) (p = 0.001), increased MCV (p = 0.014) and low RCF (p = 0.025) but no relationship with the level of serum vitamin B12 or haemoglobin. After censoring the patients with renal impairment (n = 54), the distribution of the remaining 72 patients with elevated sHcy was 37/151 (25%) with low serum vitamin B12 with or without low RCF and 35/130 (27%) with low RCF alone. sHcy correctly identified response to vitamin therapy in 33/35 (94%) patients who had adequate parameters to assess response. The positive predictive values of IF antibody/NHS, macrocytosis and/or low RCF for elevated sHcy were 100% and 34% respectively. Twenty‐four percent of patients with a low serum vitamin B12 and elevated sHcy had no abnormal haematologic parameters as determined by the routine laboratory staff. These data suggest that the usefulness of measuring sHcy in a routine diagnostic setting is limited and a careful review of the peripheral blood for macrocytosis and NHS plus determination of RCF may be a more cost‐effective process than sHcy assay in most instances to determine the presence of tissue deficiency.
Interferon treatment of refractory idiopathic thrombocytopenic purpura (ITP)
Authors: Dubbeld P. , Hillen H. F. P. , Schouten H. C.
Published: July 2010

Abstract: Up to 30% of patients with idiopathic thrombocytopenic purpura (ITP) are refractory to standard therapy with corticosteroids and splenectomy. The treatment of refractory ITP is barely effective, but promising results of interferon therapy were reported recently. In a prospective study 21 refractory ITP patients were treated with interferon alpha 2B at 3 million Units three times a week for a period of 4 weeks. Ten out of 21 patients had a response, of whom 2 had a complete response of 3.5 and 7 months duration, after which both relapsed. One other patient retreated after relapse has a complete response of 24 + months. The characteristics of the responding patients were not different from the non‐responding patients. Adverse effects were minimal. Short treatment with interferon in refractory ITP patients may be justified and the results reported here challenge us to elucidate the mechanism of action of interferon.
In vivo administration of granulocyte colony‐stimulating factor promotes neutrophil survival in vitro
Authors: Adachi Souichi , Kubota Masaru , Lin Ying Wei , Okuda Akiro , Matsubara Kousaku , Wakazono Yoshihiro , Hirota Haruyo , Kuwakado Katsuji , Akiyama Yuichi
Published: July 2010

Abstract: We recently showed that recombinant human granulocyte‐colony stimulating factor (rhG‐CSF) maintained the viability of human neutrophils in incubation for up to 72 hours. However, it is not known whether rhG‐CSF can enhance neutrophil survival in in vivo situations. To clarify this issue, we investigated neutrophil survival in vitro following in vivo injection of rhG‐CSF. Neutrophils were obtained from 4 pediatric patients with malignancies and healthy adult volunteers before and after rhG‐CSF administration. Neutrophils obtained before rhG‐CSF treatment started to undergo apoptosis after 24 h of incubation. In contrast, the survival of neutrophils drawn after rhG‐CSF administration increased by approximately 24 h. Concomitantly, the appearance of typical ladder‐like DNA fragmentation was delayed. Such an increase in neutrophil survival was inhibited by coincubation with either H 7 (10 μmol/1) or H 8 (20 μmol/1), which worked as protein kinase C inhibitors. Although our study did not measure neutrophil survival in vivo directly, it provides us with further evidence that rhG‐CSF may function to prolong neutrophil life expectancy in vivo.
Partial characterization of human leukemia U‐937 cell sublines resistant to 9‐nitrocamptothecin P
Authors: Pantazis Panayotis , Mendoza J. T. , DeJesus A. , Rubin E. , Kufe D. , Giovanella B. C.
Published: July 2010

Abstract:Human leukemia U‐937 cell sublines exhibiting various levels of resistance to 9‐nitrocamptothecin (9NC) were developed after exposure to progressively increased 9NC concentrations. Increases in 9NC resistance of the cells were accompanied by decreases in proliferation rate; appearance of morphological and functional features that correlate with granulocytic maturation; decreased synthesis of topoisomerase I; increased synthesis of topoisomerase II; and inability or decreased ability to induce tumors when xenografted in nude mice. 9NC‐resistant cells, transferred and propagated in 9NC‐free media for 6 months, continue to exhibit resistance and other features similar to cells propagated in continual presence of 9NC. Finally, 9NC‐resistant U‐937 cells respond to physiological and non‐physiological agents of cell differentiation, indicating that alternative treatments can be successfully used to inhibit growth of 9NC‐resistant U‐937 cells and tumors.
Stimulus‐specific defect in platelet aggregation in polycythemia vera
Authors: Blanc Katarina Le , Berg Anders , Palmblad Jan , Samuelsson Jan
Published: July 2010

Abstract: We have previously reported that polymorphonuclear granulocyte (PMN) and monocyte oxidative metabolism is reduced in polycythemia vera (PV) patients compared to healthy control subjects, after stimulation with cell surface receptor‐dependent stimuli such as n‐formyl‐methionyl‐leucyl‐phenylalanine, leukotriene B4 and platelet‐activating factor (PAF). In contrast, the oxidative response to phorbol myristate acetate (PMA) is normal. We now show that, in PV patients exhibiting significantly reduced PMN chemiluminescence after PAF stimulation, PAF induced platelet aggregation was also reduced — 40 ± 3% compared to 50 ± 2% in controls (p<0.01). The defective aggregatory response to PAF in PV remained over a wide range of stimuli concentrations. Platelet aggregation induced by PMA and ADP, however, was similar in PV and controls. In contrast, platelet aggregation induced by PAF (or by ADP and PMA) was not significantly reduced in patients with chronic myeloid leukemia, essential thrombocythemia and multiple myeloma. Furthermore, the release of β‐thromboglobulin was slightly but not significantly higher after PAF stimulation in PV and this argues against an abnormal PAF receptor as the cause of the defective function. Thus, not only PV neutrophils, but also PV platelets show a discrete defect of the stimulus response coupling for PAF, indicating a disease‐specific abnormality that appears to be of clonal origin.
Immunological recovery after bone marrow transplantation for severe aplastic anaemia: A Brazilian experience
Authors: de Souza Maria Helena Ornellas , Diamond Hilda Rachel , Silva Maria Luiza Macedo , Campos Mercia Mendes , Bouzas Luiz Fernando , Flowers Mary Evelyn , Rumjanek Vivian Mary , Tabak Daniel
Published: July 2010

Abstract: Twenty‐nine patients with severe aplastic anaemia (SAA) were submitted to bone marrow transplantation (BMT) and their immunological recovery analysed. Total lymphocyte counts, estimation of B lymphocytes, T lymphocytes and their subsets, natural‐killer (NK) activity were performed. Cells with the CD8 + phenotype and NK activity were the first signs of immunological recovery, whereas the CD4 + subset recovered later in patients who suffered from acute graft versus host disease (GvHD) and infections. Acute and chronic GvHD, cirrhosis, rejection and HIV viral infection contributed to the persistence of the profound immunodeficiency status observed after BMT. Our results did not differ greatly from the others and confirmed that BMT may be performed in underdeveloped countries despite the difficulties it might pose.
Effects of cyclic AMP on expression of LFA‐1, Mac‐1, and VLA‐4 and eosinophilic differentiation of a human leukemia cell line, EoL‐1
Authors: Jung Eun‐Young , Ohshima Yusei , Shintaku Noriaki , Sumimoto Shin‐Ichi , Heike Toshio , Katamura Kenji , Mayumi Mitsufumi
Published: July 2010

Abstract: We examined the effect of dibutyryl cAMP (dbcAMP) on the expression of LFA‐1 (CD1 la/CD18), Mac‐1 (CD11b/CD18), and VLA‐4 (CD49/CD29) and on eosinophilic differentiation of a human leukemia cell line, EoL‐1. Dibutyryl cAMP induced eosinophilic differentiation of EoL‐1 cells from 6–9 days after the start of culture with down‐regulation of CD 11a, CD 18, and CD49 expression and up‐regulation of CD11b expression. Changes in integrin expression, except for CD18, were seen predominantly in the fraction containing eosinophilic granule‐positive cells, suggesting that the changes were dependent on eosinophilic differentiation. On the other hand, dbcAMP‐induced changes of integrin expression were reversible and were not seen on day 9 when dbcAMP was removed on day 3, whereas eosinophilic differentiation was still present. A combination of G‐CSF and TNFα, which also induced eosinophilic differentiation of EoL‐1 cells, increased CD11b expression slightly but had no significant effect on the expression of the other integrins. Butyrate and PMA up‐regulated CD11b expression without eosinophilic differentiation. The results collectively suggest that the regulation of integrin expression on EoL‐1 cells is partly dependent and partly not dependent on eosinophilic differentiation. The possible involvement of protein kinase A and protein kinase C in these changes is suggested.
Idiopathic erythrocytosis — additional new study techniques suggest a heterogenous grou
Authors: Messinezy M. , Sawyer B. , Westwood N. B. , Pearson T. C.
Published: July 2010

Abstract: 25 patients with idiopathic erythrocytosis (absolute increase in red cell mass without conventional criteria of primary polycythaemia or known underlying cause) have been further studied for evidence of primary or secondary polycythaemia. Additional non‐conventional criteria used were: platelet distribution width, platelet nucleotide ratio, serum erythropoietin, clinical evidence of ischaemic vascular disease and erythroid culture variables in serum‐free system. All had been used in an earlier study in score form to assist in the diagnosis of primary polycythaemia. These patients were also newly assessed for the presence of hypoxia (supine oximeter values, history suggestive of sleep apnoea), for renal lesions and for splenic enlargement (impalpable) by ultrasound or computerized tomography. 7 patients had erythroid culture scores suggesting primary polycythaemia but the addition of non‐culture criteria did not result in any scores more strongly predictive of primary polycythaemia. Supine oximeter values <92% suggested hypoxaemia as the mechanism of polycythaemia in 3 patients in whom it had not previously been suspected. Some splenic enlargement (impalpable) was demonstrated in 6 patients, only 1 of whom had erythroid culture scores suggesting primary polycythaemia. 12 patients had confirmed, raised erythropoietin levels. We conclude that idiopathic erythrocytosis refers to a heterogenous group of patients. Features of primary or secondary polycythaemia may be demonstrated in some of them by additional new study techniques. The raised erythropoietin values found in half the patients were unexpected.
Revisited indications for bone marrow examinations in HIV‐infected patients
Authors: Ciaudo M. , Doco‐Lecompte T. , Guettier C. , D'Agay M. F. , David F. , Rioual N. , Buu‐Hoi A. , Molina J. M. , Kazatchkine M. , Modai J. , Lecompte T.
Published: July 2010

Abstract: We reviewed the indications for and the results of bone marrow examination (BME) from HIV‐infected patients as an attempt to improve its diagnostic yield. One‐hundred‐and‐eight bone marrow specimens from 90 patients during a 3‐year period were examined. A cytological, histological and microbiological study was carried out on the specimens. Forty‐three evaluable examinations (40% of total) performed for cytopenia showed normo‐ or hypercellularity in 33 (77%). Fifty bone marrow specimens were cultured for mycobacteria with a yield of 42% when the indication was persistent fever. Positive cultures yielded Mycobacterium avium complex in 8 out of 12 patients. Twenty‐seven patients had both culture and biopsy; granulomas were associated with all the positive (10/10) and with 1 out of 17 negative cultures (chi‐square test: p <0.001). A bone marrow involvement with lymphoma was found in 2 out of 6 patients with previously diagnosed lymphoma, and biopsy revealed a lymphoma in 2 patients. Morphological bone marrow examination should be associated with other techniques in order to appreciate bone marrow production. Bone marrow biopsy is useful for the investigation of persistent fever since granulomas suggestive of disseminated mycobacteria are frequent and allow a treatment to be initiated before microbiological confirmation and antibiotic susceptibility test.
Rapid restoration of natural killer activity following treatment with 2‐chlorodeoxyadenosine in 22 patients with hairy‐cell leukemia
Authors: Lauria F. , Rondelli D. , Raspadori D. , Benfenati D. , Tura S.
Published: July 2010

Abstract:  We report on the T‐cell profile and natural killer (NK) function in 22 hairy cell leukemia patients before and 2 and 6 months after a single 7‐day course of 2‐CdA therapy at a dose of 0.1 mg/kg daily continuous infusion. Before treatment, two groups of patients were identified in respect to NK activity, the first (12/22) had a normal number of T and NK cells and a normal NK activity, the second (10/22) showed a severe reduction of CD3 +, CD4 +, CD16 +, and CD56+ cells, together with impaired NK activity. All patients had increased serum levels of the soluble interleukin 2 receptor (sIL‐2R). Along with a high complete remission rate (77%), treatment with 2‐CdA produced a significant reduction (p = 0.001) of total lymphocytes (from 1.59 × 109/1 to 0.72 × 109/1), and particularly of CD4+ cells (from 0.68 × 109/1 to 0.23 × 109/1), while CD8+, CD16+ and CD56+ cells were less affected. Despite the dramatic reduction of lymphocytes which lasted for more than 6 months, 7/10 patients with impaired NK function before starting 2‐CdA therapy normalized the cytotoxic function within 2 months. In conclusion, 2‐CdA produces a severe reduction of lymphocytes in general, and of CD4 lymphocytes in particular, whilst it may produce a significant increase in the proportion of NK cells leading, together with a normalization of the sIL‐2R serum levels, to a rapid restoration of the NK cytotoxic function in the majority of patients.
Drug‐induced neutropenia due to direct effects on CFU‐C — Ten years of culture experience
Authors: Irvine Alexandra E. , French Anne , Daly Ann , Ranaghan Lisa , Morris T. CM.
Published: July 2010

Abstract:  Seventeen patients with suspected drug‐induced neutropenia were referred to our laboratory for investigation within a 10‐year period. In each case, the suspected drugs were incorporated separately into in vitro cultures of the patients' bone marrow. The cultures were performed in triplicate, using multiple controls. In 10 of these patients a drug‐induced inhibition of CFU‐C was demonstrated in vitro. The in vitro culture technique is a valuable investigation in patients with suspected drug‐induced neutropenia, as it can help identify the causative agent, especially in cases of multidrug administration. It also has a useful application in allowing the clinician to predict drugs to which a patient may be unduly sensitive, and prescribe accordingly.
Reactive oxygen species of neutrophils from patients with monosomy 7 in the bone marrow: Contradictory chemiluminescence activity by whole blood or by purified cells
Authors: Ristola M. , Kere J. , Ruutu T. , Repo H.
Published: July 2010

Abstract:  Monosomy 7, a deletion of the long arm of chromosome 7, was shown in the neutrophils of peripheral blood in 5 of 6 patients with a myelodysplastic syndrome or leukemia who had monosomy 7 in their bone marrow cells. In a chemiluminescence assay the production of reactive oxygen species by neutrophils from patients was increased in whole blood and decreased in purified cells, which suggests that neutrophils with monosomy 7 tolerate poorly the cell purification procedures used. The in vitro migration of purified neutrophils obtained from patients with monosomy 7 was impaired. It is known that patients with monosomy 7 have an increased susceptibility to infections. It is possible that neutrophils with monosomy 7 are too easily triggered to a full‐scale respiratory burst and thereby the cells exhaust their ability to eliminate invading microbes efficiently.
Determinants of red blood cell deformability in relation to cell age
Authors: Bosch F. H. , Werre J. M. , Schipper L , Roerdinkholder‐Stoelwinder B. , Huls T. , Willekens F.L.A. , Wichers G. , Halie M. R.
Published: July 2010

Abstract:  Red blood cell (RBC) deformability was determined with an ektacytometer in fractions separated on the basis of differences in cell volume or density. Deformability was measured with ektacytometry (rpm‐scan and osmo‐scan). We studied three groups of RBC fractions: 1. By counterflow centrifugation we obtained fractions of different cell age which showed a slight decrease in mean corpuscular haemoglobin concentration (MCHC) and an increase in surface‐to‐volume (S/V) ratio in fractions with older cells. 2. By Percoll fractionation fractions were obtained which showed a pronounced increase in (MCHC) but no change in S/V ratio. 3. By a combination of both fractionation techniques, fractions were obtained which showed an increased MCHC and an increase in S/V ratio. Deformability in group 1,2 and 3 showed respectively no change, a moderate decrease and a pronounced decrease in fractions of older cells. A decline in deformability occurs during the aging process of the red blood cell. This decline in deformability in old red cells is greater than originally thought. This decline is the result of an increase in haemoglobin concentration and a second factor, probably a decrease in membrane elasticity.
Intravascular haemolysis and increased prevalence of myeloma and monoclonal gammopathy in congenital dyserythropoietic anaemia, type III
Authors: Sandström Herbert , Wahlin Anders , Eriksson Mikael , Bergström Ingmar , Wickramasinghe Sunitha N.
Published: July 2010

Abstract:  A family with congenital dyserythropoietic anaemia type III was studied. Twenty patients and 10 of their healthy siblings were clinically examined and questioned about their medical history. Blood sampling and bone marrow aspirations were also performed. Forty‐five percent of the patients reported symptoms of anaemia and 35% regularly felt weakness, fatigue, or headache. However, the majority of the patients regarded themselves as healthy. The bone marrow showed a uniform picture of erythroid hyperplasia with multinuclear erythroblasts and gigantoblasts with up to 12 nuclei. There was laboratory evidence of intravascular haemolysis and mild anaemia. We also observed a high prevalence of monoclonal gammopathy of undetermined significance (3 cases) and myeloma (1 case) among the patients.
Effects of recombinant human stem cell factor (rh‐SCF) on colony formation and long‐term bone marrow cultures (LTBMC) in patients with myelodysplastic syndromes
Authors: Soligo Davide , Servida Federica , Cortelezzi Agostino , Pedretti Donata , Uziel Lilli , Morgutti Marina , Pogliani Enrico , Deliliers Giorgio Lambertenghi
Published: July 2010

Abstract:  Stem cell factor (SCF), the ligand of the c‐kit receptor, is a potent enhancing cytokine for haematopoietic cells in the presence of IL‐3, GM‐CSF and erythropoietin (Epo). In the clonogenic assays of 63 MDS patients, the addition of rh‐SCF + GM‐CSF and/or IL‐3 induced a significant increase (p<0.001) in the number and size of CFU‐GM. Never reaching the levels of controls, this increase was seen in all FAB subtypes, but particularly in RA. There was no significant increase in cluster formation, even in RAEB or RAEBt. Rh‐SCF (10 ng/ml) led to mean increases of up to 26 times in the number of Epo‐dependent BFU‐E colonies, particularly in RA (p<0.001) and RAEB (p<0.05). Individual responses varied widely (especially in RA) from no response to supranormal levels. Added to the weekly refeed of 37 MDS LTBMC, SCF (10 ng/ml) induced only a 7% mean increase in both cell output and the number of clonogenic cells recovered in the supernatant. Immunohistochemical examination of the supernatant showed significant increases in differentiating myeloid cells in all examined cases, and in erythroid cells in 3 cases; blast cells increased in only 3 cases. These data suggest that rh‐SCF is capable of at least partially reversing defective MDS myeloid haematopoiesis, and leads no overt risk of leukaemic transformation. Its potent effect on erythroid cells is encouraging for future clinical applications in patients, particulary if they are selected by means of in vitro tests.
Two novel human B‐cell lymphoma lines of lymphatic follicle origin: Cytogenetic, molecular genetic and histopathological characterisation
Authors: Knuutila S. , Klefström J. , Szymanska J. , Lakkala T. , Peltomäki P. , Eray M. , Teerenhovi L. , Elonen E. , Franssila K. O. , Kaartinen M.
Published: July 2010

Abstract:  Two B‐cell lines, designated as HF‐1 and HF‐4, were characterised. The cell lines have complicated karyotype abnormalities including a 14;18 translocation and an 8q24 breakpoint originating from t(2;8)(p11;q24) (HF‐1) or t(1;8)(p21:q24) (HF‐4). The lines have BCL2 rearrangement and they are positive for CD19, CD20, CD22, CD39. HF‐1 is also positive for IgG, and HF‐4 is positive for IgM and IgD. On Northern blot analyses, the 2.6‐kb and 4.2‐kb transcripts corresponding to the major transcripts of CMYC and BCL2, respectively, were seen. In Western blot as well as in FACS (fluorescence‐activated cell sorting) analysis the presence of BCL2 protein in the both HF‐1 and HF‐4 cells was demonstrated. The cell lines are expected to serve as an important tool in the study of the chromosomal mechanism activating cellular oncogenes, the somatic hypermutation mechanism of antigen‐activated B cells and the apoptosis of B cells.
A new system that analyzes erythropoietin‐mediated early signal transduction: Transfection of the c‐fos enhancer·promoter‐luciferase gene into a murine erythroid cell line
Authors: Tsuda H. , Huang R. W. , Konishi M. , Mizukami T. , Yoko‐o Y. , Hasegawa M. , Sakaguchi M. , Kawakita M. , Takatsuki K.
Published: July 2010

Abstract:  Erythropoietin (Epo) exerts its effects by binding specific receptors on the surface of reactive cells. However, the signal transduction system after binding has not been well described. To develop a system to analyze the steps of signal transduction, we transfected the human c‐fos‐enhancer/promoter linked with the Photinus pyralis luciferase gene (pfosluc2) into a murine erythroleukemia cell line ELM‐I‐1, in which we previously showed that c‐fos mRNA is rapidly induced upon Epo‐stimulation. A stable transfectant was obtained. The cells transfected with pfosluc2 were stimulated with Epo and luciferase activity in the cells was measured as light intensity. The light intensity integrated for 2 min (LI2.0) was 3202 ± 80 unit/1.5 × 105 cells before stimulation. This increased up to 5869 ± 321 unit/1.5 × 105 cells by incubating the cells with 5 U/ml Epo for 2 h. After Epo stimulation, light intensity began to increase at 30 min, reached a peak (about 1.8 times the basal level) at 120 min, and then gradually dropped. The effect of Epo was dose‐dependent; significant action occurred at as low as 0.5 U/ml, with a maximum at 5 U/ml. A similar response was observed when the cells were stimulated with interleukin‐3 (IL‐3) although the response was apparently lower than that with Epo. It was also found that IL‐3 had an additive action with Epo on c‐fos activity in this system. Thus, the above method was proven to be simple, rapid and sensitive enough to use to determine the early phase of signal transduction of Epo.