Background and objectives This study aims to investigate the possible effects of acute citrate administration on bone metabolism in healthy men.Materials and methods A placebo‐controlled, crossover trial was conducted on 10 male volunteers. The volunteers received either a standardized infusion of citrate at 1·5 mg/kg body weight/min or the equal volume of placebo, separated by a washout period of 14 days. Serial blood and urine samples were collected and analysed for bone biochemical markers and electrolytes.Results Infusion of citrate resulted in increased serum levels of the bone formation marker osteocalcin (OC) and bone resorption marker C‐telopeptide of type 1 collagen (CTX). Increases in CTX and OC were positively correlated to the surge in the serum concentration of intact parathyroid hormone (iPTH) but only OC showed correlation to changes in ionized calcium. Citrate infusion showed no effect on serum concentrations of bone alkaline phosphatase, osteoprotegerin, and bone tartrate‐resistant acid phosphatase 5b, or the expression of receptor activator of nuclear factor kappa B ligand. Variations in OC and CTX were short‐term as both bone markers gradually declined within 90 min following citrate exposure.Conclusion Acute citrate load resulted in profound alterations of the bone markers OC and CTX. The short‐term increase of CTX suggests a temporary shift to a higher bone turnover rate, although the clinical consequence of the observed changes in bone markers remains open.

Mobilized allogeneic PBPC are increasingly used instead of BM for allogeneic stem cell grafting. Although the short-term safety profile of recombinant human (rh)G-CSF seems acceptable, only minimal data on long-term safety are available. We therefore reviewed data on 171 sibling donors (M/F: 98/73) with respect to side effects of rhG-CSF and PBPC collection and impact on quality of life (QoL) and health status. In a cross-sectional study, we investigated the actual QoL and health status of the donors as well as the need for medical treatment since PBPC donation by a questionnaire that was sent to 151 donors. Ninety-five (64%) of the addressed donors responded to the questionnaire, but only 69 (46%) of them reported on their actual health status and QoL, which was good to very good in the majority of them. Two donors developed malignancies in the post-donation course. In general, PBPC collection after rhG-CSF mobilization was well tolerated by the responding donors. Although the reported events in medical history after PBPC donation do not seem to be associated with rhG-CSF administration or the collection procedure, a lifelong follow-up of donors should be obligatory.

Background In this study, we investigated the quality of autologous mononuclear cells (MNC) collected with two different cell separators using standard MNC‐apheresis procedure modalities. MNCs were purified by density gradient centrifugation and cultured according to standard protocols to generate dendritic cells (DC) and 1 × 10 7 /ml immature DCs were pulsed with tumour lysate for 3 days and subsequently characterized by fluorescent‐activated cell sorter analysis. Results No difference was found in the monocyte content of either apheresis product ( P = 0·07) and in the overall yield of MNCs ( P = 0·7). Mature DCs as defined by their phenotype revealed also no significant difference: Amicus‘, 118 × 10 6 cells ± 91 vs. AS.TEC 204®, 128 × 10 6 cells ± 137 ( P = 0·55), respectively, although the contamination with platelets (threefold) and red cells (twofold) was significantly higher in the AS.TEC 204 group ( P < 0·05) than in the Amicus group. Conclusion The Amicus and the AS.TEC 204 are equally capable in providing MNCs for the generation of DCs and the amount of concomitantly collected red cells and platelets had no impact on the final DC yield.

Background: In about 20% of allogeneic stem cell transplantation (SCT) minor ABO incompatibility does exist. This is accompanied with the risk of severe hemolysis especially during hematopoietic regeneration. The risk of hemolysis increases if peripheral blood stem cells are used instead of bone marrow (BM) and a graft-versus-host disease (GvHD) prophylaxis without methotrexate (MTX). As we have observed lethal hemolyses in patients undergoing minor ABO mismatched SCT following reduced intensity conditioning (RIC) we decided to perform prophylactic red blood cell exchange (RBCX) in all consecutive patients undergoing RIC prior to SCT with a minor ABO incompatible donor.Materials and Methods: From April 1999 to March 2006, 112 consecutive patients underwent SCT with a sibling (n=55) or unrelated donor graft (n=57) after RIC. GvHD prophylaxis consisted of CyA and mycophenolate mofetil (MMF). Fifty-four patients had an ABO-identical donor, 33 received an ABO-minor/bidirectional and 25 an ABO-major incompatible graft, respectively. RBCX was performed between day -5 and -1, removing 1 time patients RBC mass to reduce his remaining RBCs to about 40%. Packed RBCS of group 0 were used as exchange fluid.Results: So far, 20 patients underwent prophylactic RBCX before SCT. The procedure was tolerated well in 16 (80%) patients, 2 developed citrate toxicity and 2 nausea and hypotension during treatment leading to discontinuation of RBCX in the latter patients. Whereas 18 patients did not experience any signs of hemolysis after minor ABO incompatible SCT, 2 showed a marked increase in lactic dehydrogenase and serum bilirubin levels without a drop of hematocrit levels below 28% during hematologic recovery. Six of 20 patients with prophylactic RBCX died between 4.3 and 29.2 months after SCT of GvHD and infection (n=3), invasive aspergillosis (n=1) and relapse of hematologic disease (n=2). One patient rejected his graft within 1 month. The other 13 (64%) patients are in continuous complete remission a median of 13.9 months (1.2 to 36) after SCT.Conclusion: RBCX is a save and well tolerated procedure to prevent severe hemolysis by elimination of the majority of incompatible recipient RBCs as demonstrated by our data. Whether a more intensive immunosuppression can overcome the risk of hemolysis without affecting the establishment of donor chimerism remains unclear. Nevertheless, close monitoring of patients at risk between days 5 and 15 after SCT is recommended. Background: In about 20% of allogeneic stem cell transplantation (SCT) minor ABO incompatibility does exist. This is accompanied with the risk of severe hemolysis especially during hematopoietic regeneration. The risk of hemolysis increases if peripheral blood stem cells are used instead of bone marrow (BM) and a graft-versus-host disease (GvHD) prophylaxis without methotrexate (MTX). As we have observed lethal hemolyses in patients undergoing minor ABO mismatched SCT following reduced intensity conditioning (RIC) we decided to perform prophylactic red blood cell exchange (RBCX) in all consecutive patients undergoing RIC prior to SCT with a minor ABO incompatible donor. Materials and Methods: From April 1999 to March 2006, 112 consecutive patients underwent SCT with a sibling (n=55) or unrelated donor graft (n=57) after RIC. GvHD prophylaxis consisted of CyA and mycophenolate mofetil (MMF). Fifty-four patients had an ABO-identical donor, 33 received an ABO-minor/bidirectional and 25 an ABO-major incompatible graft, respectively. RBCX was performed between day -5 and -1, removing 1 time patients RBC mass to reduce his remaining RBCs to about 40%. Packed RBCS of group 0 were used as exchange fluid. Results: So far, 20 patients underwent prophylactic RBCX before SCT. The procedure was tolerated well in 16 (80%) patients, 2 developed citrate toxicity and 2 nausea and hypotension during treatment leading to discontinuation of RBCX in the latter patients. Whereas 18 patients did not experience any signs of hemolysis after minor ABO incompatible SCT, 2 showed a marked increase in lactic dehydrogenase and serum bilirubin levels without a drop of hematocrit levels below 28% during hematologic recovery. Six of 20 patients with prophylactic RBCX died between 4.3 and 29.2 months after SCT of GvHD and infection (n=3), invasive aspergillosis (n=1) and relapse of hematologic disease (n=2). One patient rejected his graft within 1 month. The other 13 (64%) patients are in continuous complete remission a median of 13.9 months (1.2 to 36) after SCT. Conclusion: RBCX is a save and well tolerated procedure to prevent severe hemolysis by elimination of the majority of incompatible recipient RBCs as demonstrated by our data. Whether a more intensive immunosuppression can overcome the risk of hemolysis without affecting the establishment of donor chimerism remains unclear. Nevertheless, close monitoring of patients at risk between days 5 and 15 after SCT is recommended.

INTRODUCTION:The aim of this investigation was to provide evidence that leukocyte depleted whole blood meets the requirements for transfusion of the European Council and thus may be an alternative to leukocyte and plasma depleted packed red blood cells in autologous blood predeposit for patients undergoing elective surgery programs.MATERIAL AND METHODS:Standard units of 450mL blood were collected from 25 healthy male volunteers. Leukocyte depletion was done via inline filtration 4h after collection. Storage lesion was assessed by measuring the release of K(+), LDH, free hemoglobin, and lactate into the storage medium, as well as by the increase of hemolysis, the decrease of pH and consumption of glucose over a storage period of 35 days. As surrogate marker for red cell quality the intracellular concentrations of adenine nucleotides [ATP, ADP, AMP] were determined.RESULTS:The extent of storage lesion remained within the ranges of standard liquid storage conditions. Hemolysis was far below the threshold of 0.8% in all WB units at the end of their shelf life. Only minor changes of intracellular adenine nucleotide levels were measured indicating a preserved function of red blood cells in leukocyte depleted whole blood. At the end of shelf life 70%+/-18% of initial ATP levels were detected.CONCLUSION:Based on our data we propose that leukocyte depleted whole blood, stored for 35 days can be an option in the autologous blood supply as it meets the requirements for transfusion of the European Council.

BACKGROUND: Allogeneic transplantation in elderly patients requires a dose-reduced conditioning regimen. Owing to reduced-intensity conditioning, host- and donor-type immune responses may affect the early post-transplant period, whereas only later on donor-derived reactions may ensue. Mismatches in the HLA system are known to be detrimental for the outcome of transplantation. Mismatches between donor and recipient for human platelet antigens (HPAs) may also affect the success of transplantation owing to serving as minor histocompatibility antigens and therefore rendering recipients at risk for graft-versus-host disease (GVHD) or graft rejection and inhibition of thrombopoiesis attributed to platelet (PLT) antibodies.PATIENTS AND METHODS: Therefore, the occurrence of GVHD, incidence of relapse, need of PLT support, and outcome by analysis of 45 donor-recipient pairs for HPA-1, -2, -3, and -5 allotypes and screening for PLT antibodies were evaluated before transplantation and again 1 year thereafter.RESULTS: Mismatches within the HPA system were not associated with an increased occurrence of transplant-related mortality or GVHD, the onset of thrombopoiesis, the frequency of PLT transfusions, or the incidence of relapse. Neither were settings of homozygous donors versus heterozygous recipients (graft-vs.-host direction) nor homozygous recipients versus heterozygous donors (host-vs.-graft direction) associated with any adverse effects on the outcome of the transplantation.CONCLUSION: Thus, the HPA match does not affect the outcome of transplantation after reduced-intensity conditioning.

Editor—The preparation of autologous blood components is regulated by rules that are derived from the production of homologous blood components. Whole blood units (WB) are stored and transfused as leucocyte- and plasma-depleted red blood cell concentrates (PRBCs) in order to minimize adverse reactions in the transfused patient. Adverse reactions caused by the plasma fraction are mainly a problem in the homologous setting. We found that leucocyte-depleted whole blood units (LD WB) stored for 35 days meet the quality requirements for transfusion of the European Council1Guide to the Preparation, Use and Quality Assurance of Blood Components. 8th edn. Council of Europe Press, Strasbourg2002Google Scholar and is a feasible option, in addition to PRBCs, for autologous transfusion, especially in hospitals without a blood bank. Standard units (450 ml) of WB were collected from 12 healthy male donors (age range 28–59 yr) using bags with an integrated filter system (MacoPharma, Toucoing, France), containing 70 ml CPDA-1 as anticoagulant/preservative solution. All units were kept at room temperature for at least 4 h to ensure bacterial phagocytosis and were then leucocyte depleted by inline filtration (Lucile, MacoPharma, Toucoing, France). Storage erosion was evaluated by the increase of K+, LDH, free haemoglobin (fHb), haemolysis rate, and the course of intracellular ATP content over the storage period of 35 days as described previously.2Leitner GC Neuhauser M Weigel G Kurze S Fischer MB Hocker P Altered intracellular purine nucleotides in gamma-irradiated red blood cell concentrates.Vox Sang. 2001; 81: 113-118Crossref PubMed Scopus (35) Google Scholar As required by the European Council, leucocyte contamination was less than 1×106 cells per unit, determined by flow cytometry (FACScan, Becton-Dickinson, San Jose, CA, USA) using the Leuco-Count kit (Becton-Dickinson) and the median Hct was 38.8% (range 34–46.9%). Haemolysis, reflected by the increase of K+, LDH and fHb is limited to 0.8% at the end of shelf life.1Guide to the Preparation, Use and Quality Assurance of Blood Components. 8th edn. Council of Europe Press, Strasbourg2002Google Scholar In LD WB units a haemolysis of 0.15%±0.11 was measured on day 35 that is clearly below the threshold. Values for K+, LDH and fHb were within the ranges observed in PRBCs (Table 1).2Leitner GC Neuhauser M Weigel G Kurze S Fischer MB Hocker P Altered intracellular purine nucleotides in gamma-irradiated red blood cell concentrates.Vox Sang. 2001; 81: 113-118Crossref PubMed Scopus (35) Google Scholar The mean (sd) intracellular content of ATP rose within 14 days from 112 (27) pmol·106 erythrocytes−1 to 146 (44) pmol·106 erythrocytes−1 and decreased thereafter. At the end of shelf life, 67 (20)% of the initial value was found. As described previously, the 24 h recovery of transfused PRBCs correlates strongly with red cell quality3Hess JR Hill HR Oliver CK et al.Twelve-week RBC storage.Transfusion. 2003; 43: 867-872Crossref PubMed Scopus (58) Google Scholar and is significantly affected when ATP concentrations fall below 10% of initial values.4Nagy S Paal M Koszegi T Ludany A Kellermayer M ATP and integrity of human red blood cells.Physiol Chem Phys Med NMR. 1998; 3: 141-148Google ScholarTable 1Metabolic parameters, increase of haemolysis, and ATP content in CPDA-1 preserved whole blood. Values are expressed as mean (sd)Days02135K+ (mmol litre−1)3.8 (1.2)19.9 (2.5)24.3 (2.6)LDH (U litre−1)166 (60)175 (56)182 (67)fHb (mg dl−1)15.7 (19.2)27.8 (22.4)30.3 (22.5)Hemolysis (%)0.07 (0.09)0.13 (0.1)0.15 (0.11)ATP (pmol·106 erythrocytes−1)112 (27)114 (28)73 (17) Open table in a new tab Erythrocytes stored as LD WB fulfil the requirements for transfusion warranted by the European Guidelines and the in vitro assessed adequate quality of erythrocytes allows the assumption that the 24 h post-transfusion recovery will not be impaired by the storage of erythrocytes as LD WB. Thus storage and transfusion of LD WB is an option in the autologous setting.

Previously, the human high mobility group protein member HMGA2 mRNA was reported to be expressed in peripheral blood of patients with breast cancer, but not in healthy individuals. Expression of HMGA2 in blood was suggested to be an independent indicator of poor prognosis in metastatic breast cancer. These very promising findings propose HMGA2 as a potential marker for the detection of circulating tumor cells in peripheral blood. Therefore, we analyzed peripheral blood specimens from healthy controls and patients with breast tumors for HMGA2 expression using TaqMan real-time RT-PCR to test if HMGA2 is a suitable marker for the early detection of breast cancer and monitoring therapy response in peripheral blood. Furthermore, we examined the possible involvement of HMGA2 expression in invasion investigated by an in vitro invasion assay using established breast cell lines. HMGA2 expression was detected in peripheral blood of breast cancer patients as well as of healthy individuals. No significant association of HMGA2 expression with any clinical or histopathological data was apparent. However, there was a significant correlation of HMGA2 expression in invasive and non invasive breast cell lines (p=0.0056). Although, HMGA2 obviously contributes to invasion it is not a specific marker for the detection of circulating tumor cells in peripheral blood.