bluuebunny/crossref_metadata_2025 huggingface dataset:
Pharmacokinetic dosing in prophylactic treatment of hemophilia A
Authors: Carlsson M. , Berntorp E. , Björkman S. , Lindvall K.
Published: July 2010

Abstract:The aim of this study was to investigate individual pharmacokinetics as a tool for dosing of factor VIII (FVIII) in severe hemophilia A. It is assumed that effective prophylaxis against bleedings is maintained if the plasma FVIII:C activity is kept above 1 U/dl, and the present study is based on this assumption. A current standard dosage regimen for FVIII is 25–40 U/kg up to three times weekly. However, there is considerable individual variation in the pharmacokinetics of FVIII:C. Individual pharmacokinetic data were used to computer‐simulate plasma activity curves after repeated doses in 8 patients. Going from prophylaxis regimens of normally 2–3 infusions per week to dosing every 2 days would theoretically reduce their average FVIII consumption by 43% with maintained or increased trough levels of FVIII:C. Daily dosing would reduce their mean FVIII usage by 82%. Modified dosage regimens, infusions every 2 days, were implemented in the patients, and plasma samples were drawn to verify the pharmacokinetic models. The feasibility of the method to generally raise trough levels with a decreased consumption of FVIII was confirmed. Dosing of coagulation factors according to kinetic principles can result in more cost‐effective utilization of these very expensive preparations.
Interferon treatment of cutaneous T‐cell lymphoma
Authors: Ross C. , Tingsgaard P. , Jørgensen H. , Vejlsgaard G. L.
Published: July 2010

Abstract:  In this report we have reviewed studies on the clinical effect of the interferon (IFN) treatment of 304 patients suffering from cutaneous T‐cell lymphoma (CTCL). Intramuscular, subcutaneous or intralesional administration of recombinant IFN has been used as monotherapy or as part of combination therapy. In general, IFN has proved to be a relatively effective agent in the treatment of CTCL, and the best responses have been achieved in the early stages of the disease. In CTCL the overall response rate to IFN including complete, partial and minor responses is 70%. Neither the doses nor the routes of administration in these studies has any statistically significant influence on the clinical response to IFN treatment. Continuous low‐dose IFN therapy, presumably in combination with psoralen and UVA light (PUVA), is recommended. This review concludes that the clinical stage of disease before treatment is the only known predictive parameter concerning the clinical response to IFN treatment in patients with CTCL.
IFN‐γ and TNF‐α secretion by CD4+ and CD8+ TCRαβ+ T‐cell clones derived early after allogeneic bone marrow transplantation
Authors: Bruserud Øystein , Hamann Wilfried , Patel Sarita , Ehninger Gerhard , Schmidt Helmut , Pawelec Graham
Published: July 2010

Abstract:  Secretion of the potentially antileukaemic cytokines IFN‐γ and TNF‐α was investigated for CD4 + and CD8 + TCRαβ+ T‐cell clones derived from 4 leukaemia patients 3–6 weeks after allogeneic BMT. We investigated cytokine secretion in response to the activation signal accessory cells + phytohaemagglutinin + Interleukin 2. All clones derived after BMT were capable of IFN‐γ and TNF‐α secretion, and both for CD4+ (n = 96) and CD8+ (n = 8) T cells quantities of IFN‐γ and TNF‐α were significantly correlated with one another. When comparing the overall results for posttransplant and normal T‐cell clones derived from 2 bone marrow donors (n = 65), both CD4+ and CD8+ TCRαβ+ T‐cell clones showed increased IFN‐γ production, and CD4+ but not CD8+ clones showed a decreased TNF‐α secretion. The results suggest that noncytotoxic T cells derived after allogeneic BMT can produce IFN‐γ and TNF‐α and may thus be capable of mediating antileukaemic effects.
Multiple myeloma treated with mitoxantrone in combination with vincristine and prednisolone (NOP regimen) versus melphalan and prednisolone: a phase III study
Authors: Keldsen Nina , Bjerrum Ole Weis , Dahl Inger Marie S. , Drivsholm Aage , Ellegaard Jørgen , Gadeberg Ole , Gimsing Peter , Grønvold Trude , Hansen Mogens Mørk , Hippe Erik , Lamvik Jon , Ly Bernt , Skarbøvik Alv , Talstad Ingebrigt , Thorling Karen , Wesenberg Karl ,   Finn Wisløff for the Nordic Myeloma Study Group(NMSG)
Published: July 2010

Abstract:  One‐hundred‐and‐fifty‐one patients with previously untreated multiple myeloma were allocated to treatment with either NOP regimen (mitoxantrone 16 mg/m2 and vincristine 2 mg day 1 and prednisolone 250 mg day 1–4 and 17–20) or M + P regimen (melphalan 0.25 mg/kg and prednisolone 100–200 mg/day day 1–4). Both regimens were repeated every 4 weeks and were scheduled for 1 year. Seventy‐seven patients were treated with NOP and 74 patients with M + P. No major clinical differences were recorded between the groups before treatment. Sixty percent of the patients responded (CR + PR) to NOP versus 64% to M + P (NS). The time to progression was 16 months (95% C.L. 14–51) in the NOP group versus 21 months (95% C.L. 15–27) in the M + P group (NS). The median survival was 14 months (7–21) in the NOP group and 31 months (21–43) in the M + P group (p = 0.02). NOP was significantly more toxic than M + P. Seven patients treated with NOP died due to infection and neutropenia and 1 patient died of cardiac toxicity, in contrast to 1 death due to infection and neutropenia in the M + P group. Gastrointestinal toxicity was acceptable in both groups. In conclusion, NOP was inferior to M + P as primary treatment of multiple myeloma.
Cadmium‐induced changes of antioxidant and metabolic status in red blood cells of rats: in vivo effects
Authors: Kostić M. M. , Ognjanović B. , Dimitrijević S. , Zikić R. V. , ZSCtajn A. , Rosić G.L. , Zivković R. V.
Published: July 2010

Abstract:  Chronic exposure of adult rats to dietary untake of cadmium (15 mg CdCl2/day/kg for 30 days) leads to development of anemia and thrombocytosis. Anemia is characterized by significant reticulocytosis (13.1 ± 1.0%), anysocytosis, poikilocytosis, iron deficiency and marked alterations of antioxidant and metabolic status of red blood cells. Activities of SOD, catalase, GPx and GR were significantly increased in red blood cells of cadmium‐treated rats. In treated animals cadmium induced an increase of red cell reduced and oxidized glutathione with no changes of GSSG/GSH ratio. However, significant reduction of lipid peroxidation was found. Plasma levels of tocopherol and ascorbate, as well as activity of glutathione‐S‐transferase, were all significantly increased in cadmium‐treated rats. The energy metabolism of red blood cells was deeply altered in cadmium‐treated rats. The levels of ATP, ADP, AMP and TAN were significantly increased while ATP/ADP ratio and adenylate energy charge (AEC) were significantly reduced. The level of 2,3‐BPG was somewhat lower, but 2,3‐BPG/Hb ratio was considerably higher, in red blood cells of cadmium‐treated rats.
Fludarabine in patients with advanced and/or resistant B‐chronic lymphocytic leukemia
Authors: Zinzani P. L. , Lauria F. , Rondelli D. , Benfenati D. , Raspadori D. , Bocchia M. , Gozzetti A. , Cavo M. , Cirio T. M. , Zaja F. , Russo D. , Fanin R. , Galieni P. , Algeri R. , Fiacchini M. , Tura S.
Published: July 2010

Abstract: In this study, we evaluated the efficacy of fludarabine (FLU), an adenine nucleoside analogue, in 35 previously treated patients with advanced and progressed B‐cell chronic lymphocytic leukemia (B‐CLL) and in 6 at diagnosis. All patients were treated at a dose of 25 mg/m2per day for 5 consecutive days (mean number of courses was 5, with a range from 4 to 6). The majority of patients experienced a beneficial effect on hematological parameters. In particular, a remarkable reduction of lymphocyte count together with an increase of neutrophils and platelets was observed. The overall response rate was 42% with 1 complete response and 16 partial responses. Ten patients achieved minor responses and the remaining 14 showed no benefit from treatment. An increased response rate was achieved in 6 untreated patients who showed an overall response rate of 67% (4/6). The major complications observed were neutropenia (66%) and febrile episodes (44%) that were generally infection‐related and were fatal in 3 cases. Because we were dealing with patients whose disease was advanced and/or resistant to treatment, the overall results may be considered encouraging with acceptable toxic reactions not superior to those frequently observed with polychemotherapy.
Corticosteroid is not beneficial in multiple‐drug combination chemotherapy for multiple myeloma
Authors: Palva I. P. , Ala‐Harja K. , Almqvist A. , Elonen E. , Hallman H. , Hänninen A. , Ilvonen M. , Isomaa B. , Jouppila J. , Järvenpáá E. , Järventie G. , Kilpi H. , Koistinen P. , Koivunen E. , Kátká K. , Kááriáinen M. , Lahtinen R. , Laitinen A. , Lehtinen M. , Mäkelä H. , Nyländen P. , Nyman D. , Oivanen T. , Pelliniemi T‐T. , Pulli T. , Rajamäki A. , Remes K. , Rosengård S. , Ruutu T. , Soininen K. , Timonen T. , Wasastjerna C. , Vilpo J. , Volin L.
Published: July 2010

Abstract:  In a randomised multicentre trial a combination of methylprednisolone, vincristine, lomustine, cyclophosphamide and melphalan (MOCCA) was compared with the same regimen omitting methylprednisolone after the first course (COLA) in previously untreated patients with multiple myeloma. The MOCCA arm showed a response rate of 72% among 79 patients and the COLA arm a response rate of 60% among 59 patients. This difference was not statistically significant. The median survival time was 56 months in the MOCCA arm and 61 months in the COLA arm. There was a slight increase of early deaths (within the first 6 months) in the MOCCA arm as compared with the COLA arm. We conclude that, in multidrug therapies, the continuation of corticosteroid at conventional dosage beyond the first course does not improve response rate or survival time in multiple myeloma.
Correlation between serum level of soluble L‐selectin and leukocyte count in chronic myeloid and lymphocytic leukemia and during bone marrow transplantation
Authors: Zetterberg Eva , Richter Johan
Published: July 2010

Abstract:  L‐selectin is a glycoprotein which is one of three members in a family of cell adhesion molecules called selectins. L‐selectin is present in distinct forms on both neutrophil granulocytes and lymphocytes, and it appears to play an important role in the early stages of leukocyte‐endothelial cell interaction. Activation of leukocytes leads to shedding of the extracellular part of L‐selectin which thus forms a soluble adhesion molecule, sL‐selectin, which retains functional capacity and can be detected in serum. In the present study we have developed a specific, sensitive sandwich ELISA to measure the serum level of sL‐selectin in patients with hematological and infectious disorders. Three patients with acute myeloid leukemia in remission and 1 patient with chronic myeloid leukemia in chronic phase were followed during bone marrow transplantation and the level of sL‐selectin was found to correlate closely to the leukocyte counts with no detectable sL‐selectin during periods of severe leukopenia. In 11 patients with chronic phase chronic myeloid leukemia and 13 patients with chronic lymphocytic leukemia the sL‐selectin level was also found to correlate closely to the leukocyte count (R = 0.98; p = 0.001 and R = 0.83; p = 0.004 respectively). One CML patient with a leukocytosis of 385 times 109/1 was found to have an sL‐selectin concentration 625 times above normal. Ten patients with acute pneumonia were evaluated at diagnosis and at the time of follow‐up 4–8 weeks later. In all patients the initial sL‐selectin level was higher than at follow‐up. However, no close correlation between sL‐selectin and leukocyte count or CRP (C‐reactive protein) at the time of diagnosis was found. In summary, we have found that the sL‐selectin level in human serum closely correlates to the leukocyte count in both CML and CLL and during bone marrow transplantation. –
Interleukin 4 inhibits the proliferation and promotes the maturation of human leukemic early B cells
Authors: Ouaaz Fateh , Mentz Franz , Mossalayi M. Djavad , Schmitt Christian , Michel Ariane , Debre Patrice , Guillosson Jean‐Jaques , Merle‐Beral Hélène , Arock Michel
Published: July 2010

Abstract:  The effects of interleukin 4 (IL‐4) on human leukemic precursor B‐cell lines were investigated. Recombinant IL‐4 (rIL‐4) was added to three acute lymphoblastic leukemia‐derived pre B‐cell lines: Reh, Km3 and Nalm‐6. Our results show that rIL‐4 significantly decreases continuous proliferation of Reh and Km3 cells while Nalm‐6 cells have a limited response in this respect. This rIL‐4 effect is dose‐dependent and can be neutralized by anti‐IL‐4 monoclonal antibody (mAb). Furthermore, rIL‐4 down‐regulated IL‐3‐induced proliferation of Reh cells. Phenotypic analysis of rIL‐4‐treated cells points to significant induction of surface marker maturation of leukemic cells by this cytokine. Together, these in vitro data suggest that IL‐4: 1) inhibits the proliferation and 2) promotes the differentiation of certain human leukemic B‐cell precursors.
Evidence that the extracellular N‐terminal domain of C5aR contains amino‐acid residues crucial for C5a binding
Authors: Mery Laurence , Boulay François
Published: July 2010

Abstract:  The human C5a anaphylatoxin is a cationic 74 amino‐acid long glycopeptide which derives from proteolysis of the fifth component of complement. It interacts with high affinity with a receptor that belongs to the G protein‐coupled receptor superfamily. Several studies have previously suggested that multiple contact points between C5a and the receptor are required to achieve high‐affinity interaction. However, at the receptor level little is known about the sites of interaction with C5a. We have investigated by in vitro mutagenesis whether the N‐terminal extracellular sequence of the C5a receptor, which is rich in aspartic acid residues, could play some role in C5a binding. Conversion of Asp10 into asparagine did not impair the level of expression at the plasma membrane, nor did it alter the affinity for C5a. However, we consistently observed a discrepancy between an apparent high level of surface expression and a weak capacity to bind C5a with high affinity, suggesting that many receptor molecules, although present on the cell surface, might be misfolded and unable to bind C5a. Replacement of Pro9 by an isoleucine had little effect, if any, on either the affinity or the C5a‐binding capacity, whereas the conversion of Pro36 into leucine dramatically reduced the expression of high‐affinity receptor at the cell surface. N‐glycosylation of human C5a receptor was found to be dispensable for the function of the receptor.
Cytoskeletal regulation of chemotactic receptors: Molecular complexation of N‐formyl peptide receptors with G proteins and actin
Authors: Jesaitis Algirdas J. , Klotz Karl‐Norbert
Published: July 2010

Abstract: Signal transduction via receptors for N‐formylmethionyl peptide chemoattractants (FPR) on human neutrophils is a highly regulated process. It involves direct interaction of receptors with heterotrimeric G‐proteins and may be under the control of cytoskeletal elements. Evidence exists suggesting that the cytoskeleton and/or the membrane skeleton determines the distribution of FPR in the plane of the plasma membrane, thus controlling FPR accessibility to different proteins in functionally distinct membrane domains. In desensitized cells, FPR are restricted to domains which are depleted of G proteins but enriched in cytoskeletal proteins such as actin and fodrin. Thus, the G protein signal transduction partners of FPR become inaccessible to the agonist‐occupied receptor, preventing cell activation. We are investigating the molecular basis for the interaction of FPR with the membrane skeleton, and our results suggest that FPR, and possibly other receptors, may directly bind to cytoskeletal proteins such as actin.
Phospholipases and activation of the NADPH oxidase
Authors: McPhail Linda C. , Qualliotine‐Mann Diane , Agwu David E. , McCall Charles E.
Published: July 2010

Abstract:  The signal transductional mechanisms regulating the activation of NADPH oxidase, the respiratory burst enzyme in phagocytic cells, are not completely understood. Receptors for most physiologic stimuli trigger the activation of various phospholipases, including phospholipases A2, C, and D. The lipid mediators formed (arachidonic acid, 1,2‐diacylglycerol, and phosphatidic acid) have been implicated as second messengers in the induction of the respiratory burst. In intact cells, we have correlated phospholipase D activation and the production of phosphatidic acid with the activation of NADPH oxidase, using the drug propranolol. Phosphatidic acid activated NADPH oxidase in a cell‐free system, but the level of activation was low. 1,2‐Diacylglycerol markedly enhanced NADPH oxidase activation by phosphatidic acid. The synergistic effect required the diacyl species, since mono‐ or tri‐acylglycerols were ineffective. Phosphatidic acid could be replaced by either lysophosphatidic acid or phosphatidylserine, but not by phosphatidylcholine, phosphatidylethanolamine, or phosphatidylinositol, suggesting specificity for an anionic phospholipid. Since other cell‐free activators of NADPH oxidase (arachidonic acid, sodium dodecyl sulfate) are also anionic amphiphiles, phosphatidic acid may directly interact with an enzyme component(s). The targets for phosphatidic acid and diacylglycerol in the cell‐free system are currently under investigation. These results emphasize the critical importance of phospholipases, particularly phospholipase D, in the regulation of the respiratory burst.
Proton channels, plasma membrane potential, and respiratory burst in human neutrophils
Authors: Demaurex Nicolas , Schrenzel Jacques , Jaconi Marisa E. , Lew Daniel P. , Krause Karl‐Heinz
Published: July 2010

Abstract: When confronted with invading microorganisms, neutrophils undergo a number of nearly synchronous reactions including the generation of microbicidal reactive oxygen intermediates by the NADPH oxidase. These reactions are accompanied by a slow depolarization, from resting values of – 60 mV to levels probably exceeding 0 mV. The depolarization is transient, indicating that a compensatory charge transport mechanism is activated. Activation of the oxidase system causes a massive burst of metabolic acid generation that would, if uncompensated, lower the intracellular pH of neutrophils by over 5 units, to lethal levels (pH = 2). Neutrophils must therefore possess particularly effective regulatory systems to avoid excessive cytosolic acidification. The recently described H+conductance of neutrophils may counteract both the acidification and the depolarization. Activation of the H+conductance occurs at depolarizing voltages and is promoted by cytosolic acidification, a combination that takes place during the respiratory burst. The NADPH oxidase of neutrophils is thus associated to an unusual, particularly efficient mechanism of H+export and charge compensation. The sequential activation of these two systems causes neutrophils to depolarize through the activation of an electron transport chain, and to repolarize through the activation of a H+conductance.
Regulation of phagocyte function by low molecular weight GTP‐binding proteins
Authors: Bokoch Gary M.
Published: July 2010

Abstract:  The mechanisms used by phagocytic leukocytes in the process of bacterial killing are regulated by GTP‐binding proteins of the Ras superfamily. In particular, the formation of toxic oxygen metabolites via the NADPH oxidase requires the action of both Rac and Rap1A proteins. Rac2 forms a third cytosolic component of the human neutrophil NADPH oxidase. Rac2 is active in its GTP‐bound form, and requires post‐translational processing (isoprenylation) in order to interact with regulatory proteins which stimulate the exchange of GTP for GDP. In the resting neutrophil, Rac is localized to the cytosol in the form of a complex with a GDP dissociation inhibitor (GDI) protein. Upon cell activation, this complex is disrupted to enable Rac to translocate to the active oxidase at the plasma membrane. The Rac‐GDI complex may be regulated by the release of specific lipids known to be generated during phagocyte activation.
Teicoplanin in Gram‐positive infection: microbiological aspects
Authors: Spencer R. C.
Published: July 2010

Abstract:  The widespread use of indwelling catheters and successful antibiotic treatment of Gram‐negative infections has led to an increase of Gram‐positive infections in severely neutropenic patients; Staphylococcus epidermidis is predominant in these infections. The problems associated with the use of vancomycin in treating such infections can be overcome by the glycopeptide antibiotic teicoplanin. It is as effective as vancomycin, but does not cause “red man” syndrome, is uncommonly nephrotoxic, can be given as a rapid bolus once daily, and routine serum monitoring is not required. Other approaches to reducing catheter‐related infections include improved training of personnel in catheter insertion and the development of new materials and methods in cannula production.
A prospective randomized study of prophylactic teicoplanin to prevent early Hickman catheter‐related sepsis in patients receiving intensive chemotherapy for haematological malignancies
Authors: Lim S. H. , Smith M. P. , Machin S. J. , Goldstone A. H.
Published: July 2010

Abstract:  In all, 88 patients with haematological malignancies requiring Hickman catheters for intensive chemotherapy were randomized to receive either one single bolus intravenous injection of teicoplanin, 400 mg, or no teicoplanin immediately before insertion of a double‐lumen Hickman catheter. Lower incidences of catheter‐related Gram‐positive sepsis were recorded in patients receiving prophylactic teicoplanin; exit site infection, tunnel infection and catheter‐related Gram‐positive septicaemia were all reduced. The benefit of prophylactic teicoplanin was observed particularly among patients who were already neutropenic at the time of catheterization. All Gram‐positive organisms isolated from infected skin sites or from blood cultures taken from Hickman catheters were susceptible to teicoplanin. No adverse reaction was reported in any of the patients receiving prophylaxis. Prophylactic teicoplanin, therefore, may be used routinely for patients requiring insertion of Hickman catheters for intensive chemotherapy, to reduce the early incidence of catheter‐related sepsis, particularly during the period of neutropenia following chemotherapy.
Teicoplanin in home therapy of the terminally ill child
Authors: Ball L.M. , Siddal S. , van Saenen H.
Published: July 2010

Abstract:  Children discharged in the terminal phase of illness were offered the possibility of having central venous line infections treated with teicoplanin at home by their parents after suitable instruction. The decision to begin antibiotic treatment was subjective, based on a history of rigors and/or raised temperature in an otherwise “well” child. No difficulties were encountered in instructing the chosen parents. In all, five treatment periods of 7 days were required in the five children selected. The review time was 31 weeks (mean duration, 6.2 weeks/patient; range, 4–12 weeks), ended in all cases by death. Infection occurred a mean of 3.2 weeks after discharge (range, 1–8 weeks), and all episodes were successfully treated at home without hospital admission or ward‐based support. No deaths occurred as a result of antibiotic therapy failure, and there were no clinically relevant side‐effects. Autopsy confirmed the absence of central venous line infection in one patient, but blood culture was positive for Staphylococcus aureus in another. This study shows that home treatment of line infections with teicoplanin is effective and well tolerated, and offers advantages in terms of quality of life and parent‐child relationships.
Teicoplanin versus vancomycin in the empirical treatment of febrile neutropenic patients
Authors: Chow Anthony W. , Jewesson Peter J. , Kureishi Amar , Phillips Gordon L.
Published: July 2010

Abstract:  Gram‐positive infections have become prevalent among neutropenic patients with cancer. A prospective, randomized, double‐blind trial of teicoplanin, 6 mg/kg every 12 h for three doses then every 24 h, versus vancomycin hydrochloride, 15 mg/kg every 12 h, in the empirical treatment of febrile neutropenic patients was undertaken among 50 consecutive patients with haematological malignancy. The patients also received piperacillin sodium, 3 g every 4 h, and tobramycin sulphate, 1.5–2 mg/kg every 8 h. Both groups (25 teicoplanin and 25 vancomycin) were comparable in age, sex, renal function, underlying disease and concurrent therapy. Among 22 patients (44%) with culture‐proven infection, Gram‐positive organisms were isolated in 15 (9 with bacteraemia) and Gram‐negative in 11 (4 with bacteraemia). Mixed or polymicrobial infection occurred in 8 patients. Serum 1‐h peak and trough levels at steady state were 41 ± 15 and 12 ± 3 mg/l for teicoplanin (at 14 ±4 days), and 40 ± 10 and 8 ± 5 mg/l for vancomycin (at 0.9 ± 0.6 days). Mean elimination half‐life and apparent volume of distribution at steady state were 80.5 ± 21.5 h and 1.4 ± 0.8 l/kg for teicoplanin, and 5.6 ± 1.8 h and 0.6 ± 0.2 l/kg for vancomycin. Empirical antimicrobial therapy was successful in 23 teicoplanin and 21 vancomycin patients, respectively (p = 0.67; two‐tailed Fisher's exact test). Nephrotoxicity (serum creatinine > 110 mmol/l), however, was more common among vancomycin patients (10 versus 2; p = 0.02), while termination of treatment due to adverse effects was also more common among vancomycin patients (10 versus 2; p = 0.02). Concurrent treatment with cyclosporin A and vancomycin, but not with cyclosporin A and teicoplanin, resulted in significant renal dysfunction (p = 0.02). At the dose employed, teicoplanin was tolerated better than vancomycin in the empirical treatment of fever and neutropenia.
Teicoplanin in combination: role in the management of the febrile neutropenic patient
Authors: Davies J.
Published: July 2010

Abstract:  The spectrum of microorganisms responsible for infection in neutropenic patients has changed in the last decade, with Gram‐positive organisms now predominant. New antibiotic strategies have been required and have evolved to cope with this change. In particular, the optimal timing of glycopeptide antibiotic use has been addressed. Both teicoplanin and vancomycin are highly effective agents for susceptible Gram‐positive organisms. Teicoplanin has advantages in terms of ease of administration and lower toxicity, while vancomycin has proved durable over many years' use. There are now firm indications for the use of glycopeptide antibiotics in febrile neutropenic patients, including clinically defined central venous catheter infection and microbiologically documented infection with susceptible organisms. The empirical use of the glycopeptide antibiotics in combination with other agents as first‐line treatment, however, is less certain. The utility of this approach remains controversial and ultimately depends on the extent to which local practice favours the emergence of Gram‐positive infections in neutropenic patients.
Mobilization of granules in neutrophils from patients with myeloproliferative disorders
Authors: Borregaard Niels , Kjeldsen Lars , Sengeløv Henrik
Published: July 2010

Abstract: Neutrophil granule subsets and dynamics were studied in 4 patients with polycythemia vera/myelofibrosis and 2 patients with chronic myelogenous leukemia. Alkaline phosphatase, a marker for the membrane of secretory vesicles (the most readily mobilizable pool of intracellular membranes in neutrophils) was highly elevated in the PV/MF patients and significantly reduced in the CML patients. In spite of this, the amount of secretory vesicles was normal as judged by the content of albumin, and of the membrane protein cytochrome b_245 and CD11b, both partially localized in secretory vesicles. Gelatinase granules were present in all patients. The azurophil granules were lighter than normal in both CML patients. SDS‐PAGE protein profiles indicated absence of defensins from azurophil granules from 1 CML patient. In addition, a 41–42 kD doublet protein band was absent from 2 PV and 1 CML patient, and reduced in the other CML patient. No difference in mobilization of granules was observed between patient neutrophils and control neutrophils. Also, stimulation with 10 8 mol/l N‐formyl‐methionyl‐leucyl‐phenylalanine induced normal increases in intracellular Ca2+ in patient neutrophils. These results indicate that stimulus‐response coupling leading to granule exocytosis is intact in neutrophils from patients with myeloproliferative disorders.
Characterization of adenosine deaminase (ADA)‐negative B‐lymphoblastoid cells cocultured with ADA‐positive fibroblasts
Authors: Fujita Masahiro , Ito Kazuhiko , Kawamoto Hiroshi , Kashii Saburo , Norioka Mihoko , Monden Sumie , Okuma Minoru
Published: July 2010

Abstract: A cell line, BAD05, derived from B lymphocytes of an adenosine deaminase (ADA; EC 3,5,4,4)‐deficient patient could not proliferate in a serum‐free medium containing 100 μmol/l deoxyadenosine. When BAD05 was cultured with ADA‐positive fibroblasts, the proliferation of BAD05 was improved. BAD05 cell density increased when the initially mixed ratio of fibroblasts/BAD05 was 1/10 or higher, but decreased when the ratio was 1/20 or lower. Deoxyadenosine concentrations in the medium and ATP and deoxyATP (dATP) levels in the BAD05 were measured after 4 hours of coculture at initial BAD05 cell densities of 1 × 105and 1 × 106cells/ml. Deoxyadenosine concentrations in the medium decreased as the density of fibroblasts increased. The dATP level decreased as the mixed ratio rose. The ratio of fibroblasts/BAD05 rather than the cell density of fibroblasts had a larger effect on the dATP levels in BAD05. Under our experimental conditions, ADA‐negative cells proliferated well when the ratio of ADA‐positive cells/ADA‐negative cells was over 1/10.
Circadian variation in serum cortisol and circulating neutrophils are markers for circadian variation of bone marrow proliferation in cancer patients
Authors: Abrahamsen Jenny Foss , Smaaland Rune , Sandberg Sverre , Aakvaag Asbjørn , Lote Knut
Published: July 2010

Abstract: Serum cortisol, circulating white blood cells and DNA cell cycle distribution in bone marrow cells were measured during daytime (11.00) and at midnight (24.00) over single 24‐hour periods in 15 cancer patients. The neutrophils and fraction of bone marrow cells in S‐phase showed the same circadian variation as cortisol with higher values in daytime as compared to midnight in 11 patients with a normal cortisol rhythm (p<0.05). The lymphocytes, eosinophils and basophils all had significantly higher values at midnight as compared to daytime. There were significant correlations between cortisol and neutrophils, lymphocytes, eosinophils and basophils. The correlation between neutrophils and fractions of bone marrow cells in S‐phase and S + G2/M‐phase were highly significant (r = 0.74, p = 0.0001 and r = 0.72, p = 0.0001, respectively). In 8 of 13 patients (61.5%) without bone marrow infiltration both cortisol and neutrophils showed identical circadian variation as bone marrow cells in S‐phase and S + G2/M‐phase. Furthermore, for the total series a significant correlation between S‐phase, cortisol and neutrophils was found by multiple regression analysis (p<0.0001). These findings strengthen the possibility of using the circadian variation in cortisol and neutrophils as marker rhythms for circadian variation in bone marrow proliferation, thus allowing optimization of cytotoxic therapy and individualization of chronotherapy.
Mononuclear cells from peripheral blood of untreated hairy‐cell leukaemia patients enhance the growth of BFU‐e
Authors: Nielsen Bendt , Hokland Peter
Published: July 2010

Abstract: To further characterise the regulation of haematopoiesis in hairy‐cell leukaemia (HCL), we investigated the effect of mononuclear cells from peripheral blood (PBMNC) of untreated HCL patients on the in vitro growth of BFU‐e. The effect was tested in an autologous system employing post‐treatment HCL PBMNC, and in an allogeneic system. A significant enhancing effect of the pre‐treatment HCL PBMNC was seen. In contrast to studies by others, IFN‐α consistently inhibited the growth of BFU‐e from normal donors, from pre‐ and post‐treatment HCL PBMNC, and mixtures thereof. Since PBMNC from patients with very high percentages of circulating hairy cells (HC) also had an enhancing effect, HC was a likely source of one or more enhancing factors. Thus, even though we cannot exclude a minor inhibitory activity of the HC by our assay, their net effect seems to be enhancing, and therefore it seems unlikely that the HC is directly responsible for the anaemia in HCL.
Metabolic evidence of cobalamin deficiency in bone marrow cells harvested for transplantation from donors given nitrous oxide
Authors: Carmel Ralph , Rabinowitz Arthur P. , Mazumder Amitabha
Published: July 2010

Abstract: Nitrous oxide inactivates cobalamin, but clinically apparent sequelae ensue in nondeficient individuals only when exposure is prolonged. The gas is widely used in anesthesia, therefore, and is commonly given to donors during harvesting of their bone marrow cells for transplantation. The present study shows that nitrous oxide administered for only 75–120 minutes induced mild but unequivocal DNA synthetic abnormalities attributable to cobalamin deficiency in the harvested marrow cells of 4 out of 5 donors; the deoxyuridine suppression test in these 4 patients showed abnormal results more than 4 standard deviations above the reference mean. Metabolic evidence of cobalamin deficiency in cryopreserved cells diminished only slightly when they were thawed and retested 1 day later, but was no longer detectable in cells thawed and tested on the 3rd day. In contrast, cells harvested under nitrous oxide‐free anesthesia in 4 subjects showed no evidence of cobalamin deficiency in the deoxyuridine suppression test. These results demonstrate that even relatively brief exposure to nitrous oxide induces cobalamin deficiency in harvested bone marrow cells, and that the cells remain metabolically impaired for more than 24 hours. Although clinical sequelae are not apparent at this time, the several potential implications of our findings indicate that the use of nitrous oxide in bone marrow transplantation needs to be evaluated further.
Phospholipid determination in platelets, plasma and red cells of patients with chronic myeloproliferative disorders
Authors: Cesar Jesús M. , Vecino Ana , Perez‐Vaquero Mikel , Navarro José L.
Published: July 2010

Abstract: Red cell phospholipids (PLs) were assessed in 11 patients with essential thrombocythemia and 5 patients with polycythemia vera. Platelet and plasma PLs were also determined in 10 of these patients, and the results were compared with studies performed in 16 healthy volunteers. The amount of platelet PLs in patients was similar to controls (556 ± 90 nmol/109cells, versus 481 ± 91 nmol/109cells), as was the percentage of the main specimens of these compounds, including phosphatidylserine (11.1 ± 0.8%), which is relevant for platelet procoagulant activity. We did not find differences between red cell PLs of patients (300 ± 60 nmol/109cells), versus controls (289 ± 71 nmol/109cells), and the sphingomyelin/phosphatidylcholine ratio in these cells was the same in both groups (0.75 ± 0.1). Finally, we did not detect any alteration in the amount of plasma PLs specimens.
Myelosuppression in HCL: Role of hairy cells, T cells and haematopoietic growth factors
Authors: Schwarzmeier J. D. , Gasché C. G. , Hilgarth M. F. , Reinisch W. W. , Göbl S. , Berger R.
Published: July 2010

Abstract: To elucidate mechanisms which may be responsible for the haematopoietic insufficiency in hairy cell leukaemia (HCL), we investigated in an autologous in vitro system the influence of haematopoietic growth factors (CSFs) and the effects of hairy cells (HCs) as well as T cells on the formation of haematopoietic colonies (CFU). Colony forming assays were performed using peripheral blood mononuclear cells (PBMC) of 6 HCL patients. To remove HCs, PBMCs were subjected to complement‐mediated lysis, T cells were removed by E‐rosette formation. Assays were done with and without recombinant human (rh) interleukin‐3 (IL‐3) and rh granulocyte‐macrophage‐colony‐stimulating factor (GM‐CSF). All 6 patients exhibited a severe reduction of their circulating progenitor cell (CPC) compartment. There was no correlation between the degree of colony reduction and the number of HCs. However, a correlation was found between the numbers of CPCs of HCL patients and healthy donors and the monocyte counts in these groups (r = 0.8573, p<0.001). The removal of autologous HCs, but also of T cells, resulted in a significant increase in colony formation (BFU‐E, CFU‐GM, CFU‐mix). In none of the experiments, however, did colony numbers come close to the normal range. This was only achieved by supplementation of the culture medium with rh IL‐3 and rh GM‐CSF. The results suggest that the haematopoietic failure observed in HCL patients is probably due to an inadequate supply of CSFs as well as to an inhibitory activity of HCs and T cells which might exert their effects in a synergistic fashion. There is also evidence that the lack of monocytes plays a role in the development of the haematopoietic insufficiency in HCL.
Magnetic activated cell sorting (MACS) — a new immunomagnetic method for megakaryocytic cell isolation: Comparison of different separation techniques
Authors: Schmitz B. , Radbruch A. , Kümmel T. , Wickenhauser C. , Korb H. , Hansmann M.L. , Thiele J. , Fischer R.
Published: July 2010

Abstract: Megakaryocytes are difficult to isolate because of their fragility, their tendency to aggregate, and their varying sizes. For purification of cells at different stages of maturation and of different sizes (ploidy classes) we developed an immunomagnetic cell sorting method (MACS) to enrich the whole spectrum of the megakaryocytic cell lineage. The use of small magnetic beads coupled to various antibodies and labelling with fluorescent antibodies allowed direct analysis of enrichment and evaluation of the isolated fraction without further staining or detachment procedures. CD 61 (Y2/51), a monoclonal antibody directed against platelet glycoprotein IIIa, was employed to perform the separation procedure. An enrichment up to 47% of CD 61‐positive cells with an average of 37% and a recovery rate of 37% was obtained by using the MACS technique. Pre‐enrichment by Percoll density centrifugation, followed by MACS separation, resulted in an enrichment of 65% and a recovery rate of 67%. The relative amount of small megakaryocytic cells in only MACS‐enriched cell populations, however, was higher than in Percoll/MACS fractions. As a parameter of vitality we tested cytokine secretion of the enriched megakaryocytes in reverse haemolytic plaque assays. Secretion of IL‐1, IL‐6, GM‐CSF, and PDGF with and without stimulation by phorbol myristate acetate was demonstrable at the single cell level.
Haemochromatosis as a window into the study of the immunological system: A novel correlation between CD8+ lymphocytes and iron overload
Authors: Porto Graça , Reimαo Raquel , Gonçalves Cristina , Vicente Corália , Justiça Benvindo , de Sousa Maria
Published: July 2010

Abstract: In the present study we report a serial investigation of the numbers of the peripheral blood cells — erythrocytes, polymorphonuclear neutrophils, total lymphocytes, T‐lymphocyte subpopulations (CD2, CD4, CD8), B lymphocytes and monocytes — in a group of 21 patients with haemochromatosis during the time of intensive phlebotomy treatment, i.e., from iron overload until the onset of iron deficiency. A remarkable individual stability of all blood cell populations studied was found in all patients. Patients differed in their relative proportions of CD4+ and CD8+. Each individual's CD4/CD8 ratio, as well as the absolute numbers, remained unaffected with time, confirming the existence of a strict homeostatic regulation of the relative numbers of the two major peripheral T lymphocytes. A significant positive correlation between CD4/CD8 ratios and the amount of iron mobilised by phlebotomy was found during this study. A novel correlation between the relative proportions of CD4+ and CD8+ cells and iron overload is confirmed by the follow‐up of iron re‐entry in the serum transferrin pool in the treated patients.
Hydroxyurea enhances 3′‐azido‐3′‐deoxythymidine (AZT) cytotoxicity in human chronic myeloid leukemia models*
Authors: Tosi Patrizia , Visani Giuseppe , Ottaviani Emanuela , Tura Sante , Darnowski James W.
Published: July 2010

Abstract:In this report we have evaluated the cytotoxic activity of 3′‐azido‐3′‐deoxythymidine (AZT) used in combination with hydroxyurea (HU), an agent which disruptsde novothymidylate synthesis. In 2 chronic myeloid leukemia (CML) cell lines, K.562 and RWLeu4, the IC50 of AZT was 8 (μmol/l and 28 μmol/l respectively, after a 5‐day exposure, and the IC50 of HU was 80 μmol/l and 70 μmol/l respectively. In the presence of various concentrations of HU (1 μmol/l‐100 μmol/l) the IC50 of AZT in both cell lines was significantly reduced and subsequent isobologram analysis revealed synergistic activity. Similarly, analysis of [3H]AZT incorporation into the DNA fraction of these cells indicated that exposure to AZT + HU resulted in an increased incorporation of AZT into DNA when compared to incubation in AZT alone. Biochemically, this effect appeared to be related to a decrease in dTTP pools caused by HU. The combination AZT + HU has also been demonstrated to exert a synergistic effect in inhibiting colony growth of bone marrow granulocyte‐macrophage progenitors (CFU‐GM) from patients affected by Ph1+CML in chronic phase. These results are promising in view of a possiblein vivoutilization of this drug combination.
Expression of adhesion molecules in endothelial cells during allogeneic bone marrow transplantation
Authors: Shen Naizhi , Ffrench Patrick , Guyotat Denis , Ffrench Martine , Fiere Denis , Bryon Paul‐André , Dechavanne Marc
Published: July 2010

Abstract: Endothelial cell activation during allogeneic bone marrow transplantation, mainly in acute graft‐versus‐host disease (aGvHD) was studied in 23 recipients and 5 controls using anti‐von Willebrand factor (vWF) antibody, antibodies to endothelial leukocyte adhesion molecule‐1 (ELAM‐1), vascular cell adhesion molecule‐1 (VCAM‐1) and intercellular adhesion molecule‐1 (ICAM‐1), and anti‐HLA‐DQ antibody, by immunohistological staining of skin. vWF extravasation, ELAM‐1 and VCAM‐1 expression were present in most recipients with a cutaneous rash which was confirmed as an aGvHD by histological examination (documented aGvHD) (p = 0.005 for vWF extravasation and ELAM‐1 expression and p = 0.03 for VCAM‐1 expression in comparison with the controls). In recipients with a rash, the cases displaying vWF extravasation and ELAM‐1 expression were significantly more numerous in those with a documented aGvHD than in those without histological features of aGvHD (p = 0.01). vWF extravasation and ELAM‐1 occurred concomitantly (p<0.01). This study demonstrates that, during the course of skin aGvHD following bone marrow transplantation, there is transient expression of ELAM‐1 and VCAM‐1 by endothelial cells and simultaneous vWF extravasation, indicating an intense inflammation with endothelial cell participation.
High‐dose cyclophosphamide for mobilization of circulating stem cells in chronic myeloid leukemia
Authors: Tringali Stefania , Santoro Alessandra , Scimé Rosanna , Vasta Stefania , Pampinella Maria , Marino Maria Assunta , Majolino Ignazio
Published: July 2010

Abstract: Experimental and clinical data suggest that Ph‐negative myeloid progenitor cells are present, albeit suppressed, in the bone marrow of chronic myeloid leukemia (CML) patients. These residual Ph‐negative cells might, in certain circumstances, regain their proliferative advantage over the leukemic Ph‐positive clone. Treating CML patients with intensive chemotherapy might allow the harvest, in the early phase of recovery, of Ph‐negative stem cells to be used as graft after myeloablative regimen. In our study, 6 CML patients were admitted to a program of autograft with circulating stem cells (CSC) collected after high‐dose (5 or 7 g/m2) cyclophosphamide (HD‐CY) mobilization. All were autografted, using busulphan 16 mg/kg and melphalan 60 mg/m2. As graft, 4 patients received CSC only, while 2 patients were also given bone marrow, as their peripheral blood CFU‐GM yield was unsatisfactory. Two previously alpha‐IFN‐responding patients showed a slow hematologic recovery, but achieved a marked and further reduction of their Ph‐positive metaphases post‐graft. Moreover, in one of them, cytogenetic analyses performed on apheresis product showed a more pronounced reduction of his Ph‐positive metaphases, as compared to bone marrow samples, suggesting a potential purging effect of the mobilization procedure.
Serum immunoreactive erythropoietin in hyper‐ and hypothyroidism: Clinical observations related to cell culture studies
Authors: Brenner B. , Fandrey J. , Jelkmann W.
Published: July 2010

Abstract:Laboratory experiments have demonstrated that tetra‐ and triiodothyronine (T4, T3) enhance hypoxia‐induced erythropoietin (Epo) production. In the present study serum immunoreactive Epo was measured in 29 patients with hyperthyroidism and in 10 patients with hypothyroidism. Epo levels were inversely correlated to the blood haemoglobin concentration [Hb] in both groups of patients. However, Epo levels at given [Hb] were significantly higher in the hyperthyroid state.In vitrostudies confirmed that T4and T3stimulate Epo synthesis in the human liver cell line HepG2. This stimulating effect persisted for at least 1 day after the removal of T4and T3from the cultures. Thus, while thyroidal disorders affect steady‐state levels of circulating Epo, it seems unlikely that thyroid hormones play a major role in abrupt adjustments of Epo production, such as the diurnal changes.
Serum ferritin in children and adolescents. Results from population surveys in 1979 and 1986 comprising 1312 individuals
Authors: Milman Nils , Backer Vibeke , Laursen Eva Mosfeldt , Graudal Niels , Ibsen Karsten Kaas , Jordal Robert
Published: July 2010

Abstract: Iron stores were assessed by measurement of serum ferritin in a population survey comprising 436 randomly selected urban Danish schoolchildren (219 boys, 217 girls) 7 to 17 years old. Median ferritin values were similar in boys and girls 7 to 13 years old, while 14‐ to 17‐year‐old boys had significantly higher values than girls of similar age (p< 0.0001). Boys had stable median ferritin values from 7 to 15 years of age, after which a significant rise was found from 16 to 17 years of age (p<0.01). Girls had unchanged median ferritin values from 7 to 11 years of age, after which a significant decrease was found from 12 to 17 years of age (p<0.01). The frequencies of low ferritin values < 16 μg/1 (indicating absent iron stores) were: Children 7 to 11 years, 1.0%. Boys 12 to 15 years, 4.5%; 16 to 17 years, 0%. Girls 12 to 15 years, 7.6%; 16 to 17 years, 16.7%. Ferritin levels in the reported 1986 survey were significantly higher than those obtained in a similar survey in the same area in 1979. The results indicate an overall increase in iron stores in Danish schoolchildren and adolescents during 1979–1986, possibly due to an increase in the consumption of meat, and in the use of vitamin‐mineral supplements containing iron.