bluuebunny/crossref_metadata_2025 huggingface dataset:
Three monoclonal IgG components, an IgG4(Λ), an IgG2(κ) and an IgG1/IgG3 (κ) Gm(f,b) hybrid, in a single myeloma patient
Authors: Milošević‐Jovčić N. , Dovezenski Nebojša , Jovanović L. , Rolović Z. , Gotić M. , Radošević N. , Suvajdžić N. , Tošić L.
Published: July 2010

Abstract: An unusual triclonal IgG combination in the serum of a 56‐year old male with clinical stage IIIB multiple myeloma is reported. The patient initially had an IgG4(Λ) monoclonal protein in his serum and later developed an IgG2(κ) and an IgG (κ) which possessed the characteristics of both IgG1 and IgG3 subclasses with an unusual combination of allotypic markers. Three M‐proteins did not share idiotypic determinants. A rare class‐switch recombination followed by mutation has been considered as a possible mechanism leading to this combination.
Priming and treatment with molgramostim (rhGM‐CSF) in adult high‐risk acute myeloid leukemia during induction chemotherapy: a prospective, randomized pilot study
Authors: Hansen Per Boye , Johnsen Hans E. , Jensen Linda , Gaarsdal Eva , Simonsen Kåre , Ralfkiaer Elisabeth
Published: July 2010

Abstract: In a randomized study of 18 adult patients with high‐risk or advanced acute myeloid leukemia (AML) we investigated the effect of supplementing conventional induction chemotherapy with recombinant human granulocyte‐macrophage colony‐stimulating factor (rhGM‐CSF). For comparison, a historical control group of 90 patients treated for de novo AML with conventional chemotherapy during the previous period, 1984–1990, was also analyzed. Before induction chemotherapy, 10 patients were randomized to receiving rhGM‐CSF, starting on day 1 to 3 before chemotherapy and continued for a maximum of 21 days after the start of induction treatment. Fatal complications and treatment outcome did not differ between the study groups and historical controls. Nor were there any differences between the groups in terms of hematological toxicity, e.g. time to three‐lineage regeneration and need for supportive therapy. However, sequential weekly bone marrow examinations revealed a prolonged reduction of the relative number of myeloid (CD33‐positive) marrow cells in the rhGM‐CSF treated group. Although the small number of patients studied may not permit a definite conclusion, this randomized study did not demonstrate major beneficial effects of combining rhGM‐CSF with standard induction chemotherapy in high‐risk patients with AML.
Computerised method for recording platelet density distribution
Authors: Järemo Petter
Published: July 2010

Abstract: In the present study a computerized apparatus was employed for scanning light transmission variations along test tubes containing density‐separated platelets. The device consists of a stepping motor, a stationary halogen lamp and a photopotentiometer connected to a personal computer. Anticoagulated whole blood was layered on a preformed continuous Percoll gradient having a density span from 1090 kg/l (bottom) to 1040 kg/l (top). After centrifugation at 3400g for 1.5 hours, high‐density cells (i.e. erythrocytes) pass through to the bottom of the test tube and the lighter platelets remain in the gradient. The test tube is moved by the computer between the halogen lamp and the photopotentiometer. Transmission variations along the gradient were recorded and registered in the computer. Density markers beads were used as an internal standard and platelet peak density was determined. After perforating the test tube the gradient was divided into 45 aliquots. In all fractions determination of platelet counts and mean platelet volume was carried out. In addition, in the aliquots having a platelet count > 20 × 1012/l the ratio β‐thromboglobulin per platelet was also determined. The platelet distribution in the gradient was illustrated graphically. A good agreement was found when comparing platelet distributions in the gradients and light transmission variations along the test tubes.
Inversions of the factor VIII gene in Swedish patients with severe haemophilia A
Authors: Ljung Rolf , Sjörin E.
Published: July 2010

Abstract: The series comprised 49 Swedish patients with severe haemophilia A [belonging to 49 families (21 with known and 28 with sporadic haemophilia)], of whom 12 had developed F. VIII inhibitors. Using Southern blotting, 45% (22/49) were found to have inversions, i.e., intrachromosomal rearrangements of the tip of the X‐chromosome. Twenty patients had one or the other of the two variants of inversions recently published, whereas 2 patients manifested novel band patterns. Inversions were found in 50% of the families with sporadic haemophilia, and in 38% of those with known haemophilia. Fourteen families with sporadic haemophilia A had inversions, the proband carrying the de novo mutation in 4 cases and the proband's mother in 10 cases. Six inversions derived from a male and five from a female X‐chromosome meiosis, the origin of the remaining three was not established. Genetic counselling of patients with severe haemophilia A and their families will be considerably improved, as inversions occur in half the severe cases and can be detected by a simple Southern blotting procedure.
Simultaneous measurements of megakaryocyte‐associated IgG (MAIgG) and platelet‐associated IgG (PAIgG) in chronic idiopathic thrombocytopenic purpura
Authors: Nagasawa Toshiro , Hasegawa Yuichi , Komeno Takuya , Itoh Takayoshi , Ninomiya Haruhiko , Abe Tsukasa
Published: July 2010

Abstract: We have simultaneously measured platelet‐associated IgG (PAIgG) and megakaryocyte‐associated IgG (MAIgG) in 30 untreated patients with chronic idiopathic thrombocytopenic purpura (CITP). Megakaryocytes were purified from bone marrow by 35% Percoll gradient centrifugation, followed by negative immunopanning using magnetic immunobeads. The normal range of MAIgG in 30 healthy donors was 15.5 & 10.0 ng/105 megakaryocytes, whereas MAIgG in the 30 CITP patients was 140 & 59.3 ng/105 megakaryocytes, although the values were widely distributed. From the PAIgG and MAIgG data, CITP patients were classified into three types; type I (PAIgG < 200 ng/107 platelets and MAIgG < 150 ng/105 megakaryocytes), type II (PAIgG > 200 ng and MAIgG > 150 ng), and type III (PAIgG < 200 ng and MAIgG > 150 ng). Patients with types I and III had good clinical courses, but, in contrast, patients with type II responded poorly to steroid therapy followed by splenectomy or became refractory to treatment. In splenectomized patients, MAIgG of responder was promptly decreased to normal range and, in contrast, that of non‐responder was persistently elevated. These results indicate that anti‐platelet autoantibodies are able to bind with megakaryocytes in the bone marrow as well as with platelets in the peripheral blood, and the results also suggest that megakaryopoiesis in CITP is heterogeneous. Simultaneous measurement of PAIgG and MAIgG may predict the clinical outcome of CIPT.
Increase of monocytes predicts mobilization of peripheral stem and progenitor cells after chemotherapy followed by G‐CSF administration
Authors: Hansson Mona , Svensson Anna , Engervall Per , Björkholm Magnus , Gruber Astrid , Söderström Tommy
Published: July 2010

Abstract: Mobilization of primitive haematopoietic cells to the peripheral blood was studied in 25 patients with haematological malignancies. The optimal level of peripheral stem cells (PSC), defined by their surface expression of CD34, was significantly higher after mobilization with G‐CSF, either following chemotherapy or alone (median: 123 × 106/l and 143 × 106/l of CD34+ cells respectively) than without administration of G‐CSF subsequent to chemotherapy (median: 27 × 106/l of CD34+ cells). An individual variation in when optimal mobilization of CD34+ cells and myeloid progenitors occurs after chemotherapy and G‐CSF administration was noted (median: day 12, range 7–24 days), which makes it difficult to predict when PSC collections in a given patient should be performed. In this study, chemotherapy followed by G‐CSF administration resulted in a short lasting (2–3 days) peak appearance of CD34+ cells that could predicted by a 2‐fold increase in absolute numbers of monocytes, as compared to the previous day. After the peak level of CD34+ cells in the blood was reached, no further increase in monocytes was seen. The identification of an increase in monocytes, to be used as a predictive variable for when optimal mobilization of PSC will occur in a given patient, may be particularly useful in the individual timing of PSC collections from non‐hospitalized patients.
Correlation between soluble transferrin receptor and serum ferritin levels following bone marrow transplantation for thalassemia
Authors: Centis F. , Delfini C. , Agostinelli F. , Barbanti I. , Annibali M. , Lucarelli G.
Published: July 2010

Abstract: This study analyzes the serum transferrin receptor (sTfR) levels in a series of 230 ex‐thalassemics with a follow‐up of 1 to 9 years after bone marrow transplantation (BMT) for homozygous β thalassemia. Ex‐thalassemics are individuals, cured of homozygous β thalassemia by BMT, who maintain different degrees of iron overload acquired during the pretransplant period. Both in experimental and clinical conditions, sTfR concentrations have been shown to be a quantitative measure of body iron status. This study was carried out to assess whether the level of sTfR may be of help in determining the extent of iron overload in ex‐thalassemics. Patients who received the marrow from their HLA‐identical sibling donor heterozygous for β thalassemia, namely heterozygous ex‐thalassemics, displayed significantly higher levels of sTfR than patients transplanted from their normal sibling donors (normal ex‐thalassemics). This finding suggests that increased erythropoiesis, albeit in part ineffective in heterozygous ex‐thalassemics, is responsible for the sTfR increment. Both heterozygous and normal ex‐thalassemics had significant lower sTfR levels than their heterozygous (p < 0.003) or normal (p < 0.0001) donors, respectively. These differences may be ascribed to the presence of iron overload in ex‐thalassemics in comparison to their normal or heterozygous donors who did not present excess of iron in the body. A significant inverse correlation between sTfR and serum ferritin levels (r = –0.54, p < 0.0001) was found when normal ex‐thalassemics were considered. In heterozygous ex‐thalassemics, the lack of correlation between these two parameters may be explained by the enhanced erythropoietic activity of individuals with thalassemic trait. These results suggest that the level of sTfR may be a useful indicator of iron overload in normal ex‐thalassemics.
Effects of Erwinia‐asparaginase on the coagulation system
Authors: Carlsson H. , Stockelberg D. , Tengborn L. , Braide I. , Carneskog J. , Kutti J.
Published: July 2010

Abstract: l‐Asparaginase treatment during induction therapy in acute lymphoblastic leukaemia (ALL) is known to be frequently complicated by thromboembolic events. It was recently suggested that l‐asparaginase derived from Erwinia chrysanthemi alters the coagulation system less severely than does Escherichia coli asparaginase. In a series of 11 adult patients with ALL, we investigated some parameters of the coagulation system during treatment with Erwinia asparaginase. The doses employed were rather high; all patients below the age of 60 years received 15000 U/m2 daily over 14 days. In accordance with what is known from treatment with E. coli asparaginase, we observed significant lowering of antithrombin as well as of fibrinogen. However, as to fibrinogen indeed a significant decrease had occurred prior to the institution of Erwinia asparaginase treatment. The most striking observation in the present study was that the levels of prothrombin complex, reflecting the function of K‐vitamin dependent coagulation factors II, VII and X, remained within normal ranges during treatment. This indicates that these coagulation factors were not affected by Erwinia asparaginase, an observation at variance with several reports where E. coli asparaginase was investigated. This latter observation was the only finding which could lend support to the view that Erwinia asparaginase affects the coagulation system less than E. coli asparaginase. Finally, one of our patients developed a sinus thrombosis, a severe thrombotic complication.
Platelet ultra‐structure study in Budd‐Chiari syndrome
Authors: Dayal S. , Pati H. P. , Pande G. K. , Sharma M. P. , Saraya A. K.
Published: July 2010

Abstract: Morphologic characteristics of circulating platelets were studied in 20 patients with primary Budd‐Chiari syndrome (BCS) without any known etiology, using transmission electron microscopy (TEM). Significant platelet ultrastructural changes were observed in all the patients (in 10←90% platelets) as compared to 20 normal healthy controls (in up to 4% of their platelets). The prominent changes in the platelets were paucity or absence of alpha granules, hypertrophy of the open canalicular systems (OCS) and clumping and fusion of the granules and other organelles in the centre of platelets. Some other changes observed in platelets were dilated channels of OCS, pseudopodial protrusion of cytoplasm and presence of prominent masses of glycogen particles. Platelets from 20 normal controls processed along with the patients' platelets showed only a few such abnormalities. Most of these changes observed in patients' platelets were akin to the changes observed in platelets undergoing activation. Assay of plasma beta‐thromboglobulin showed significantly higher levels in all the patients (p< 0.001) further confirming on‐going in vivo platelet activation with morphologic changes most likely reflecting the thrombotic process present in BCS patients.
Retroviral‐mediated gene transfer into human bone marrow stromal cells: Studies of efficiency and in vivo survival in SCID mice
Authors: Li K. J. , Dilber M. S. , Abedi M. R. , Björkstrand B. , Smith C. I. E. , Garoff H. , Gahrton G. , Xanthopoulos K. G.
Published: July 2010

Abstract: Retroviral‐mediated gene transfer into bone marrow cells is extensively used in gene therapy protocols. Cytokines are needed for stimulation, to achieve a high rate of gene transfer. However, the stromal cell compartment of bone marrow is characterized by rapid proliferation even without cytokines. In this study, human bone marrow stromal cells were isolated and subsequently infected with recombinant retrovirus in a cell‐free supernatant. The LN retroviral vector used in this study carries the bacterial neomycin phosphotransferase gene (neoR). Transduction efficiency was significantly enhanced by repeated cycles of infection, with a maximum of 91% transduced cells by four rounds of infection. Presence of the neoR‐gene was detected by PCR from all stromal cells selected by G418. After culture in vitro for 3 months, cells were still positive for PCR amplification of the neoR‐gene. Transduced stromal cells were also injected into SCID mice to study their homing and survival ability in vivo.
Antibodies against platelet GPIb/IX, GPIIb/IIIa, and other platelet antigens in chronic idiopathic thrombocytopenic purpura
Authors: Hou M. , Stockelberg D. , Kutti J. , Wadenvik H.
Published: July 2010

Abstract: Antibodies involved in the pathogenesis of chronic idiopathic thrombocytopenic purpura (ITP) react most frequently with platelet glycoprotein (GP) Ib/IX and GPIIb/IIIa. However, uncertainty as to the specificity, frequency, and clinical significance of such antibodies still remains. By using a modified antigen‐capture ELISA (MACE), an immunoprecipitation assay, and an immunoblot assay, sera from 60 patients with chronic ITP were analyzed. GP‐specific antibodies were found in 50% (30/60) of the patients, with 14 patients having antibodies directed solely to GPIIb/IIIa, 8 holding antibodies specific only for GPIb/IX, and 8 possessing antibodies against both antigens. Serum antibodies were more frequently (p<0.01) detected in either active and/or non‐splenectomized ITP patients. Moreover, in patients displaying antibodies against GPIb/IX, significantly (p<0.05) lower platelet counts were observed. Using the immunoblot assay, antibodies specific for a 30 kD platelet antigen were detected in 12 of 60 patients. This antigen could not be immunoprecipitated from surface labelled platelet membranes, indicating an intracellular location. We conclude that in chronic ITP, (1) the frequency of anti‐GPIIb/IIIa antibodies is close to that of anti‐GPIb/IX antibodies, (2) anti‐GP antibodies are more likely to be detected in patients with an active disease status and, (3) a 30 kD internal platelet protein is another frequent antigen.
Identification of latent myeloproliferative disease in patients with Budd‐Chiari syndrome using X‐chromosome inactivation patterns and in vitro erythroid colony formation
Authors: Acharya J. , Westwood N. B. , Sawyer B. M. , Messinezy M. , Burroughs A. K. , Mehta A. B. , Pearson T. C.
Published: July 2010

Abstract: Some patients with an early or latent myeloproliferative disorder (MPD) present with Budd‐Chiari syndrome (BCS, hepatic vein thrombosis). Cell culture analysis of erythroid progenitors (BFU‐E) can be used to discriminate primary from secondary MPD and examination of X‐chromosome inactivation (in females) can be used to demonstrate clonality in neoplastic tissues. The present study used these techniques to examine whether a group of 7 female patients who presented with BCS had evidence to support a diagnosis of MPD. Unilateral X‐inactivation and therefore clonality can be studied in females heterozygous for X‐linked restriction fragment length polymorphisms (RFLP) by differences in methylation between active and inactive chromosomes. Probes for two polymorphic loci, phosphoglycerate kinase (PGK, at Xq13.3 [BstX1 RFLP]) and M27β (an anonymous locus DXS255 at Xp11.22 [Pst1 RFLP]) were used to study methylation patterns. All 7 patients were heterozygous using M27β and 2/7 were also heterozygous using the PGK probe. Polyclonal patterns of X‐inactivation in granulocytes were demonstrated in 3/7, a skewed/monoclonal pattern in 1/7 and aberrant patterns in 3/7 using M27β. Two patients who had aberrant patterns of X inactivation with M27β demonstrated a skewed/monoclonal pattern with PGK. The results of BFU‐E growth patterns and clonality were entirely concordant in 5/6 patients. Thus X‐chromosome inactivation patterns, in conjunction with erythroid colony studies, can be used to assist in the diagnosis of an underlying MPD in BCS.
Globin chain turnover in reticulocytes from patients with β°‐thalassaemia/Hb E disease
Authors: Kalpravidh R. W. , Komolvanich S. , Wilairat P. , Fucharoen S.
Published: July 2010

β°‐Thalassaemia/Hb E disease is the most frequent β‐thalassaemia in Thailand. However, patients have a varying degree of anaemia. The difference in severity is attributed to a differential accumulation of unpaired α‐globin chains, which is the net result of biosynthesis and catabolism. Turnover of newly synthesized globin chains in reticulocytes from β°‐thalassaemia/Hb E patients was determined. Proteolysis was ATP‐independent and degraded only 10–15% of the radiolabelled globin during a 4‐h incubation period at 37 °C and there was no difference in globin turnover in reticulocytes from subjects with mild and severe forms of β°‐thalassaemia/Hb E. These results indicate that excess α‐globin chains do not act as substrates for the reticulocyte proteolytic system and that the difference in severity of anaemia in β°‐thalassaemia/Hb E disease cannot be explained by a difference in selective post‐translational catabolism.
Increased susceptibility of microcytic red blood cells to in vitro oxidative stress
Authors: Corrons J. L. Vives , Miguel‐Garcia A. , Pujades M. A. , Miguel‐Sosa A. , Cambiazzo S. , Linares M. , Dibarrart M. T. , Calvo M. A.
Published: July 2010

Abstract: Oxidative damage to erythrocytes in thalassaemia has been related to generation of free radicals by an excess of denaturated α‐ or β‐globin chains, intracellular iron overload and low concentration of normal haemoglobin (HGB). Two good indicators of such oxidative damage are the high red blood cell (RBC) malonyldialdehyde (MDA) production detected following exogenous oxidant stress and the decrease of pyrimidine 5′‐nucleotidase (P5N), the most sensitive enzyme to SH‐group damage in vivo. Conflicting data, however, have so far accumulated in the literature concerning differences in oxidative damage between the different forms of thalassaemia and iron deficiency anaemia (IDA). In the present study, oxidative susceptibility, as defined by the production of MDA in vitro and antioxidant capacity, as measured by the activity of RBC glutathione peroxidase (GPx), superoxide dismutase (SOD) and by reduced glutathione (GSH), have been studied in microcytic RBCs from patients with β‐thalassaemia trait, Spanish (δβ)°‐thalassaemia heterozygotes (δβ‐thalassaemia trait) and iron deficiency anaemia (IDA). The results are consistent with the existence of significant differences in the severity and pattern of oxidative stress susceptibility between β‐thalassaemia trait (increased MDA production and higher SOD and GPx activities) and the other two forms of microcytosis (δβ thalassaemia trait and IDA).

Furthermore, the finding of normal P5′N activity in δβ thalassaemia trait, gives further support to the less intense peroxidative environment of RBCs in this form of thalassaemia when compared to β‐thalassaemia trait, characterized by acquired RBC P5′N deficiency due to oxidative damage.
Effects of granulocyte‐macrophage colony stimulating factor produced in Chinese hamster ovary cells (regramostim), Escherichia coli (molgramostim) and yeast (sargramostim) on priming peripheral blood progenitor cells for use with autologous bone marrow after high‐dose chemotherapy
Authors: Hussein Atif M. , Ross Maureen , Vredenburgh James , Meisenberg Barry , Hars Vera , Gilbert Colleen , Petros William P. , Coniglio David , Kurtzberg Joanne , Rubin Peter , Peters William P.
Published: July 2010

Abstract: Peripheral blood progenitor cells (PBPCs) were collected without prior association with chemotherapy but after the administration of granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) produced in Chinese hamster ovary cells (CHO‐GM, regramostim), Escherichia coli (E. coli‐GM, molgramostim), or yeast (Yeast‐GM, sargramostim) and used in conjunction with autologous bone marrow after high‐dose chemotherapy in 69 patients with breast cancer or melanoma. The mean peripheral white blood cell (WBC) counts increased by 2.2 to 2.7‐fold after regramostim, 4.5 to 7.3‐fold after molgramostim and 4.3‐fold after sargramostim. All patients underwent three leukaphereses. The mean (± standard error) total nucleated pheresed cells per kg × 108 were 4.15 ± 0.56, 15.10 ± 1.77 and 7.24 ± 1.00 for patients receiving regramostim, molgramostim or sargramostim respectively. The mean (± standard error) granulocyte‐macrophage colony‐forming units per kg × 104 mobilized into the PB were 8.75 ± 3.63, 71.03 ± 17.85, and 65.11 ± 18.74 for patients receiving regramostim, molgramostim, or sargramostim respectively. The total mean (± standard error) CD34+ cells per kg × 107 collected by three leukaphereses were 3.28 ± 1.62, 1.34 ± 0.51 and 2.57 ± 1.93, for patients receiving regramostim, molgramostim or sargramostim respectively. The use of either molgramostim‐ or sargramostim‐primed PBPCs led to complete elimination of absolute leukopenia with a WBC count under 100/mm3 in 64% and 77% of patients treated, respectively. Patients receiving molgramostim‐primed PBPCs required fewer red blood cells transfusions than patients receiving regramostim‐primed PBPCs (p = 0.0062). Our data indicate that PBPCs collected without prior association with chemotherapy but after either molgramostim or sargramostim with autologous bone marrow support and GM‐CSF shorten the hematopoietic recovery after myeloablative chemotherapy in patients with breast cancer or melanoma.
Soluble receptors for tumour necrosis factor in clinical laboratory diagnosis
Authors: Diez‐Ruiz Antonio , Tilz Gernot P. , Zangerle Robert , Baier‐Bitterlich Gabriele , Wachter Helmut , Fuchs Dietmar
Published: July 2010

Abstract:  Soluble tumour necrosis factor receptors (sTNF‐Rs) play a role as modulators of the biological function of tumour necrosis factor‐α (TNF‐α) in an agonist/antagonist pattern. In various pathologic states the production and release of sTNF‐Rs may mediate host response and determine the course and outcome of disease by interacting with TNF‐α and competing with cell surface receptors. The determination of sTNF‐Rs in body fluids such as plasma or serum is a new tool to gain information about immune processes and provides valuable insight into a variety of pathological conditions. Regarding its immediate clinical use, sTNF‐Rs levels show high accuracy in the follow‐up and prognosis of various diseases. In HIV infection and sepsis, sTNF‐Rs concentrations strongly correlate with the clinical stage and the progression of disease and can be of predictive value. Determination of sTNF‐Rs also gives useful information for monitoring cancer and autoimmune diseases. The information provided is often even superior to that obtained with classical disease markers, probably due to the direct involvement of the “TNF system” in the pathogenetic mechanisms in these patients. The available data imply that the measurement of sTNF‐Rs, especially of the sTNF‐R 75kD type, is a useful adjunct for quantification of the Th1‐type immune response, similar to other immune activation markers such as neopterin and β2‐microglobulin. Endogenous sTNF‐Rs concentrations appear to reflect the activation state of the TNF‐α/TNF receptor system.
Clinical implications of cytokine and soluble receptor measurements in patients with newly‐diagnosed aggressive non‐Hodgkin's lymphoma
Authors: Stasi Roberto , Zinzani Pier Luigi , Galieni Piero , Lauta Vito Michele , Damasio Eugenio , Dispensa Egidio , Dammacco Franco , Venditti Adriano , Del Poeta Giovani , Cantonetti Maria , Perrotti Alessio , Papa Giuseppe , Tura Sante
Published: July 2010

Abstract: Serum levels of 13 different cytokines and receptors were measured serially in 78 patients with aggressive non‐Hodgkin's lymphoma (NHL) treated by 4 cycles of an intensive multi‐agent chemotherapy regimen. Recombinant human granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) was administered subcutaneously in 36 of these patients from day + 5 to day + 18 after each chemotherapy. Statistically significantly higher pretreatment levels of interleukin‐2 (IL‐2), interleukin‐6 (IL‐6), interleukin‐8 (IL‐8), interleukin‐10 (IL‐10), the soluble IL‐2 receptor (sIL‐2r), the soluble transferrin receptor (sTf‐r), and neopterin, were observed in NHL patients as compared to controls (p< 0.001 for all molecules). sIL‐2r and sTf‐r levels correlated with tumor burden (p< 0.001 and p = 0.003, respectively) whereas IL‐6 was higher in patients presenting B symptoms (p< 0.001). Cytokine levels progressively declined to normal ranges in responding patients, while they remained elevated in non‐responders. Relapsed patients also presented increased concentrations of several molecules. During the administration of GM‐CSF, we observed the drastic increase of sIL‐2r, while lower elevations were recorded for a number of cytokines, including IL‐8, tumor necrosis factor‐α, interleukin‐1β, IL‐6, and IL‐2. However, upon completion of the induction treatment, cytokine/receptor levels were comparable among individuals with the same type of response, whether or not they had received GM‐CSF. No single parameter was found to be of prognostic significance, but the combination of elevated IL‐10 and of sIL‐2r greater than 3000 U/ml selected a subgroup of 7 patients who failed induction treatment (p = 0.002). These results demonstrate that cytokine and soluble receptor measurements can provide valuable informations for a better management of NHL, in terms both of markers to monitor disease activity and of prognostic indicators.
Outpatient supportive therapy after induction to remission therapy in adult acute myelogenous leukaemia (AML) is feasible: a multicentre study
Authors: Guillermo J. Ruiz‐Argüelles MD, FACP  , Apreza‐Molina Ma. Guadalupe , Alemán‐Hoey Deborah Dion , Gómez‐Almaguer David , Marín‐López Antonio , Mercado‐Díaz Leopoldo
Published: July 2010

Abstract:  Twenty‐four adult patients with AML were treated with standard “7 + 3”chemotherapy. After administering the myeloablative drugs in the hospital, patients were instructed to continue their supportive treatment on an outpatient basis; they received ciprofloxacin, cotrimoxasole and itraconazole vo until the absolute granulocyte count rose above 1 × 109/l. Platelet concentrates were given every other day until the platelet count rose above 20 × 109/l. Complete remission (CR) was obtained in 87%. Fever developed in 29% and 2 cases were complicated by indwelling‐catheter‐related Pseudomona aeruginosa septicaemia, 1 Entamoeba hystolytica enteritis and 1 Pneumocystis carinii pneumonia; these patients were hospitalized to treat these infections specifically. In no case was the infection fatal. The median disease free‐survival (DFS) was 17 months, 12‐month DFS was 66%, and 30‐month DFS was 17%. Our calculations have shown that 1700 USD/patient were saved by avoiding prolonged hospitalization; this may provide not only economical, but also psychological advantages to patients.
Survival and risk of leukaemia in polycythaemia vera and essential thrombocythaemia treated with oral radiophosphorus: Are safer drugs available?
Authors: Brandt L. , Anderson H.
Published: July 2010

Abstract:  For 366 patients with polycythaemia vera (PV) or essential thrombocythaemia (ET) diagnosed 1971–1990, oral administration of 32‐P was used as myelosuppressive treatment. Retreatment was not restricted to any defined interval and the number of treatments or the total dose were not limited. For 107 patients, follow‐up was > 10 years. 15 of these presented with life‐threatening occlusive vascular symptoms and their survival was short. For the remaining 92 patients 5‐yr survival was not significantly worse than for a Swedish population matched for age and sex. Survival at 10 yr was lower, 51% versus 66% expected. Acute myeloid leukaemia (AML) was diagnosed in 11 of the 107 patients (10.3%). In the whole material of 366 patients, 17 have developed AML with a median time of 8.5 yr from start of treatment. There was a maximum incidence of 4% per yr after 8–12 yr. Later, the incidence decreased. The median annual dose of 32‐P for the AML patients was 100 MBq and was not significantly larger than for a matched control group surviving without AML, 96 MBq. The results are compared with reports on PV or ET patients treated with busulphan (Bu) or hydroxyurea (HU). With comparable periods of follow‐up there are no indications that an adequate myelosuppression with oral 32‐P will be associated with shorter survival or higher incidence of AML than treatment with Bu or HU. It is concluded that, for the time being, oral administration of 32‐P is an acceptable standard treatment in PV and ET.
Changes in adhesion molecule expression and function in B‐cell chronic lymphocytic leukaemia after in vitro interferon‐α stimulation
Authors: Csanaky G. , Vass J. A. , Ocsovszki I. , Milosevits J. , Szomor Á. , Schmelczer M.
Published: July 2010

Abstract:  Peripheral blood mononuclear cells (PBMCs) from 10 B‐CLL patients were investigated after 24 hours of in vitro interferon‐α (IFN‐α) stimulation. The constitutional expression of the L‐selectins (LECAM‐1), LFA‐1/CD11a, VLA α‐4/CDw49d and ICAM‐1/CD54 adhesion molecules was detected, and changes in their density after IFN‐α stimulation were compared to results obtained by the high endothelial venule (HEV)‐binding assay and a carbohydrate (phosphonomannan core polysaccharide: PPME and fucoidin) immobilization test. The LECAM‐1 and ICAM‐1 molecules were expressed on the great majority of CLL cells, while the LFA‐1 and VLA‐4 α‐chains were expressed by only a small number of cells. Statistically significant changes (p< 0.001) were observed in LECAM‐1 antigen density (changes in mean cell fluorescence), as well as in functional tests (HEV‐, PPME‐ and fucoidin‐binding; p<0.01) after in vitro IFN‐α stimulation. Based on a prior study (Jewell et al., Leukemia 1992: 6: 400–404) and on the present findings, not only an increased expression but also an enhanced function of the L‐selectins seem to be well substantiated after IFN‐α stimulation, which may explain the therapeutic effect of IFN‐α in reducing the accumulation of leukaemic B cells in the blood. The remarkably high expression of ICAM‐1 in this series necessitates further studies to clarify the exact expression rate and role of this molecule.
Disseminated intravascular coagulation (DIC) in adult patients with acute leukaemia
Authors: Sletnes K. E. , Godal H. C. , Wisløff F.
Published: July 2010

Abstract:  In 71 patients with acute leukaemia admitted for remission induction, disseminated intravascular coagulation (DIC) was looked for in 50 patients and diagnosed in 10 (20%). Of 10 patients with acute lymphoblastic leukaemia, 3 had DIC, and of 40 patients with acute myeloblastic leukaemia, 7 had DIC. The presence of DIC was related to bleeding manifestations within the first 2 weeks. A haemorrhagic diathesis was present in all DIC patients: 4 had minor and 6 had major bleeding, i.e. WHO grade ≥ 2. In addition to blood product support, most DIC patients were treated with low doses of heparin and tranexamic acid. In all DIC patients the haemorrhagic symptoms preceded the heparin administration. Among 40 screened patients without DIC, 17 patients had minor and 3 had major haemorrhagic manifestations. Thus, the proportion of patients with major bleeding was significantly greater among the DIC patients (6/10 vs 3/40, p< 0.001). In conclusion, DIC at presentation was associated with a significantly increased risk for severe haemorrhagic complications and should be looked for in adults with acute leukaemia.
Sequential administration of recombinant human granulocyte‐macrophage colony‐stimulating factor and human erythropoietin for treatment of myelodysplasia syndromes
Authors: Runde V. , Aul C. , Ebert A. , Grabenhorst U. , Schneider W.
Published: July 2010

Abstract:  Treatment of myelodysplastic syndromes (MDS) with recombinant human erythropoietin (Epo) is successful in only 10% to 25% of patients. We performed a pilot study in 10 anaemic patients with MDS to examine whether sequentially applied recombinant human granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) and Epo improves haemoglobin levels and/or reduces red blood cell transfusion requirements. Morphological diagnoses of patients were refractory anaemia (RA) in 3 cases, RA with ring sideroblasts in 3 cases and RA with excess blasts in 4 cases. GM‐CSF was given subcutaneously at a dose of 150 μg/m2/d during the initial 10 days. From day 11, Epo was administered by subcutaneous injections for 8 weeks at a dose of 100 U/kg/d and subsequently at an escalated dose of 200 U/kg/d in 3 patients. Changes in reticulocyte counts, haemoglobin levels, RBC support and ferrokinetic parameters were compared with pretreatment values. Two out of 8 évaluable patients showed a rise in haemoglobin levels at week 8 and 10, respectively, and lost their transfusion dependency for a period of 13 and 27 weeks. In 1 patient, haemoglobin level increased only after dose escalation of Epo (200 U/kg/d). Leukocyte counts remained uneffected by treatment with Epo, while 1 patient showed a 4‐fold increase in platelet numbers. Toxicity was mild. Two patients died of pneumonia and global heart failure, respectively, unrelated to growth factor therapy. Based on this pilot study, we conclude that sequential treatment with GM‐CSF and Epo does not increase erythroid responses in anaemic patients with MDS. Because of the delayed increase in haemoglobin in both responders, we surmise that the beneficial effects were induced by Epo alone.
Thymic stromal cells support differentiation of natural killer cells from CD34+ bone marrow cells in vitro
Authors: Tjønnfjord Geir E. , Steen Rita , Veiby Ole Petter , Egeland Torstein
Published: July 2010

Abstract:  Natural killer (NK) cells are CD3− CD56+ lymphocytes characterized by exhibiting non‐MHC restricted cytotoxicity. A developmental relationship between NK cells and T lymphocytes has been proposed, and, moreover, the thymus has been shown to contain NK cell precursors. In this study we utilized an in vitro assay, devised to study T‐lymphocyte development from bone marrow progenitors, to investigate the ability of thymic stromal cells to support generation of NK cells from CD34+ bone marrow cells. CD34+ cells purified from healthy adults were seeded on adherent thymic stromal cells. The cells emerging after culture were phenotypically characterized by flow cytometry. We show that lymphocytes expressing the phenotypical characteristics of NK cells were generated from CD34+ bone marrow cells, and that these cells represented 1% of the cells recovered from the cultures. Furthermore, this was accomplished without supplement of exogenous interleukin 2 which is required for NK cell differentiation in bone marrow cultures.
Effects of interleukin‐3 following chemotherapy of non‐Hodgkin's lymphoma. A prospective, controlled phase I/II study.
Authors: Hovgaard D. J. , Nissen N. I.
Published: July 2010

Abstract: The effect of rhIL‐3 was investigated in 32 patients with newly diagnosed non‐Hodgkin lymphoma in a phase I/II trial. All patients received 6 cycles of standard CHOP chemotherapy, and each patient was his own control where rhIL‐3 was given as a daily s.c. injection for 14 days (day 2–15) in cycle 2 and 4, while cycle 1 and 3 were control cycles. Five dose levels were examined (0.5 — 1 — 5 — 7.5 — 10 μg/kg). Compared to the other more lineage‐specific hemopoietic growth factors G‐ and GM‐CSF, the effect of rhIL‐3 on the hemopoiesis was less dramatic and more delayed, i.e. the most apparent effect was observed in the 2 weeks of treatment. Thus, the neutrophil counts from days 15 to 22 following CHOP were significantly raised and the duration of neutropenia was shorter (significantly only at 10 μg/kg), while the nadir values were unaffected. Platelet recovery from days 12–22 was significantly increased and nadir values occurred earlier compared to control cycles, but were only increased in some subsets. Other cell populations affected moderately in the recovery period were eosinophils and monocytes. Reticulocytes increased, but no effect on hemoglobin or RBC transfusion requirement was noted. Only moderate adverse reactions occurred such as fever, chills, flushing of the face and flu‐like symptoms. There was no evidence of stimulation of tumor growth. Most significant, the rhIL‐3 treatment at all but the lowest dose levels led to an improved tolerance to chemotherapy, as indicated by a decline in number of delayed cycles. A conclusion concerning the role of rhIL‐3 as post‐chemotherapy adjuvant should await studies using rhIL‐3 in combination with more lineage‐restricted hemopoietic growth factors.
Proliferation and differentiation of myelodysplastic CD34+ cells in serum‐free medium: II. Response to combined colony‐stimulating factors
Authors: Sawada Ken‐ichi , Sato Norihiro , Notoya Atsushi , Tarumi Takashi , Yasukouchi Taro , Yamaguchi Miki , Kohno Michihumi , Mikuni Chikara , Koike Takao
Published: July 2010

Abstract: To investigate the role of colony stimulating factors (CSFs) in the proliferation and differentiation of progenitor cells from myelodysplastic syndromes (MDS), marrow progenitor cells from 18 MDS patients were highly purified using CD34 monoclonal antibody and immunomagnetic microspheres (MDS CD34+ cells). These cells were cultured in serum‐free medium with various combinations of five colony stimulating factors (CSFs): recombinant human interleukin‐3 (rIL‐3), granulocyte/macrophage‐CSF (rGM‐CSF), granulocyte‐CSF (rG‐CSF), macrophage‐CSF (rM‐CSF), and erythropoietin (rEP). Among the tested CSFs, such as rM‐CSF, rG‐CSF, rGM‐CSF and rIL‐3, a combination of the first three CSFs was the most effective stimulus for the proliferation of non‐erythroid MDS progenitor cells. An increase of undifferentiated “blast” cell colonies in 5/18 MDS patients occurred and these 5 patients belonged to the high‐risk group. In the presence of these three CSFs, rIL‐3 had no effect on the proliferation and differentiation of MDS CD34+ cells; however, IL‐3 was efficient for the proliferation of MDS CD34+ cells to the erythroid lineage. rGM‐CSF or rIL‐3 alone did not efficiently support proliferation and differentiation of CD34+ cells. M‐CSF is present in normal human serum at a concentration of 550 ±110 U/ml, a concentration exceeding that used in this study (100 U/ml). Therefore, in vivo administration of G‐CSF combined with GM‐CSF to MDS patients may be one of the most effective CSF combinations for proliferation of MDS progenitor cells to the non‐erythroid lineage. However, the effect on the capacity for differentiation was minimal, especially in patients belonging to the high‐risk group.
Gamma‐chain heterogeneity in Greek (δβ)O‐thalassemia.
Authors: Georgiou Ioannis , Seferiadis Konstantin , Lolis Dimitrios , Tsolas Orestes , Bourantas Kostantinos L.
Published: July 2010

Abstract: A molecular and biochemical population study of (δβ)O thalassemia in central Greece is described. The molecular study was focused on the type of the deletion and the status of Gγ‐Xmn***l polymorphism, whereas the biochemical approach was centered on the Gγ/Aγ ratio as well as the frequency of the Aγ T chain in the fetal hemoglobin of 19 δβ‐thalassemia heterozygotes and 3 homozygotes. This study includes individuals from the mountainous district of Epirus (northwestern Greece) where the trait was found to be concentrated along the river Arachthos. The Sicilian (δβ)O thalassemia deletion was found in all subjects tested by direct PCR. The levels for the Gγ‐chain presented values ranging from 29 to 83% of the total γ ‐chain content. Thirteen heterozygotes had the adult Gγ /Aγ ratio (mean Gγ: 35% ± 10) of whom 10 were Xmnl‐negative (‐/‐), 6 had the newborn ratio (mean Gγ: 70% ± 9) and were Xmnl‐positive, while homozygotes had equal amounts of Gγ and Aγ. Five of the 19 heterozygotes were Aγ T‐positive with low levels of this Aγ‐chain variant, suggesting an in‐trans to the δβ‐thalassemia determinant production.
Secretion of leukaemia inhibitory factor after allogeneic bone marrow transplantation: a study of CD4+ and CD8+ TCRαβ+ T‐cell clones derived from four leukaemia patients.
Authors: Bruserud Øystein , Hamann Wilfried , Pawelec Granham
Published: July 2010

Abstract: CD4+ and CD8+ TCRαβ+ T‐cell clones were derived from 4 leukaemia patients early (4–6 weeks) after allogeneic bone marrow transplantation. Leukemia inhibitory factor (LIF) secretion in response to the activation signal accessory cells (AC) + phytohaemagglutinin (PHA) was investigated for each individual clone. Only a minority of CD4 + TCRαβ + T‐cell clones secreted LIF in response to AC + PHA, whereas most T‐cell clones were capable of LIF secretion when exogenous interleukin 2 was added together with AC + PHA. LIF secretion could also be demonstrated for CD8+ TCRαβ+ posttransplant T‐cell clones. Thus, posttransplant CD4+ and CD8+ TCRαβ + clonogenic T‐cells are capable of LIF secretion, and LIF secretion may be a T‐cell effector mechanism in graft versus host disease, graft versus leukaemia effects or posttransplant haematopoietic reconstitution.
The metabolism of erythropoietin in liver cirrhosis patients compared with healthy volunteers.
Authors: Jensen J.D. , Jensen L.W. , Madsen J.K. , Poulsen L.
Published: July 2010

The purpose of the investigation was to study the metabolism of erythropoietin (EPO) in patients with liver disease. Twelve patients with liver cirrhosis and 10 healthy volunteers were studied. The patients were moderately anemic with a hematocrit of 33 vs 42% (medians) in the volunteers. The pharmacokinetic parameters were calculated after an intravenous (i.v.) injection of 100 U/kg of recombinant human EPO. The serum EPO was measured by radioimmunoassay at regular intervals until 48 h. The median terminal elimination half life in the cirrhosis patients was 5.15 h vs 5.37 h in the control subjects. The clearance was 7.78 vs 7.52 ml/min/1.73 m2 (ns). The steady‐state volume of distribution was 3.69 vs 3.09 1/1.73 m2 (ns). The estimated endogenous EPO production was significantly higher in liver cirrhosis (486 vs 290 U/d/1.73m2, p<0.01). The basal serum EPO was significantly higher in the cirrhosis patients (43.5 vs 26.3 U/***l, p<0.01). The hematocrit correlated inversely with the basal serum EPO level in the cirrhosis patients (r= ‐0.63, p<0.04). The EPO‐clearance was not related to the presence of ascites, esophageal varices, or to abnormal blood chemistry. It was concluded that normal metabolism of EPO was maintained in liver cirrhosis and that the cirrhotic patients had a moderate compensatory increase of EPO production in response to anemia
A new, peroral non‐radioactive vitamin B12 absorption test compared with the Schilling test
Authors: Magnus Erik , Müller Carl
Published: July 2010

Abstract: The results of a non‐radioactive, peroral absorption test have been compared with the results of the traditional Schilling test in 31 cobalamin‐deficient patients. The non‐radioactive test is simple to perform, is less costly than the Schilling test and seems to give reliable results. The non‐radioactive test should be performed after cobalamin treatment, but not until the plasma cobalamin value has declined to below 450 pmol/l. Normal Schilling test was noted in one‐third of the patients, while normal non‐radioactive test was noted in only one‐fifth of the patients. The results reveal some discrepancies between the two tests regarding the response to intrinsic factor. In the non‐radioactive test without intrinsic factor, the great variation in values may reflect varying secretion of intrinsic factor, possibly secondary to infestation with Helicobacter pylori. “False normal” Schilling test seems to be more common than previously believed.
Fludarabine treatment in resistant Waldenstrom's macroglobulinemia
Authors: Zinzani P. L. , Gherlinzoni F. , Bendandi M. , Zaccaria A. , Aitini E. , Salvucci M. , Tura S.
Published: July 2010

Abstract: Fludarabine (FLU) is a fluorinated purine analogue with a promising antineoplastic activity in lymphoproliferative disorders. In this study, we evaluated the efficacy of FLU in 12 previously treated (primary refractory and refractory relapse) patients with Waldenstrom's macroglobulinemia. All patients were treated at a dosage of 25 mg/m2 per day for 5 consecutive days for a total of six courses. Of the 12 patients, 5 (41%) achieved partial response (PR), and the remaining 7 showed no benefit from the treatment. An increased response rate was obtained in the 4 primary refractory patients in which 2 PR were documented. Treatment was well‐tolerated and there were no Fludarabine‐related fatalities. With a mean follow‐up of 10 months, only 1 PR patient has relapsed. Fludarabine is an interesting new salvage agent effective against recurrent/resistant Waldenstrom's macroglobulinemia and should be evaluated in further studies in untreated patients with Fludarabine in monochemotherapy or in combination with other active modalities.