bluuebunny/crossref_metadata_2025 huggingface dataset:
Benefit of Prestorage Leukocyte Depletion of Single‐Donor Platelet Concentrates
Authors: Chalandon Y. , Mermillod B. , Beris Ph. , Doucet A. , Chapuis B. , Roux‐Lombard P. , Dayer J.‐M.
Published: December 2004

Background and Objectives:
The presence of contaminating white blood cells (WBCs) in platelet concentrates is associated with transfusion reactions and may adversely alter the quality of platelets during storage. Leukocyte depletion by filtration of platelets has been increasingly used to avoid these complications. However, the best time for filtration and the benefits of filtering single‐donor platelet concentrates (thrombapheresis, TH) have yet to be clearly defined. Methods: In a randomized study of 202 TH collected with an Autopheresis C system, we determined whether prestorage filtration (preSF) of WBCs from TH as compared with poststorage (bedside) filtration (postSF) resulted in a better product. Levels of cytokines and C3a accumulating in the medium during storage, platelet activation state, in vivo platelet recovery, and transfusion reactions were compared in pre‐ and poststorage products. Results: As compared to preSF, significantly more postSF TH had detectable levels of tumor necrosis factor‐α (TNF‐α; 47 vs. 15%; p<0.0001) and interleukin 6 (13 vs. 3%; p = 0.02), lower pH (p<0.0001) and decreased levels of C3a (910 vs. 2,000 pg/ml; p<0.0001). Futhermore, platelet activation was increased in postSF TH (p = 0.022). PostSF TH tended to plug the bedside filter (27% of postSF TH delivered) from day 3 onward. There was also a significant difference in platelet recovery, postSF TH showing a lower corrected count increment (CCl; p = 0.0055) when taking into account the postSF TH that plugged filters (CCl = 0), but no difference when plugged TH were excluded. A correlation could be established between TNF‐α levels and poor in vivo recovery (p<0.0001). Febrile nonhemolytic transfusion reactions were low in both groups (4 and 9%). Conclusion: These results indicate a benefit of preSF TH as compared with postSF TH based on the following parameters: decrease in cytokine levels, less platelet activation, maintenance of higher pH, and more efficient use of stored platelets (27% of postSF TH were lost because of plugging of filters). These results apply particularly to the Autopheresis C systems with its high initial WBC content.
Economic Impact of Donor Platelet Count and Platelet Yield in Apheresis Products: Relevance for Emerging Issues in Platelet Transfusion Therapy
Authors: Goodnough Lawrence Tim , Ali Suhail , Despotis George , Dynis Marian , DiPersio John F.
Published: December 2004

Introduction:
We analyzed donor platelet counts and platelet product yields in 708 consecutive platelet aphereses in our program in order to define the importance of this relationship for emerging issues in platelet transfusion therapy. Methods: Aphereses performed on the Spectra 3.6 (COBE, Lakewood. Colo.) the CS‐3000 Plus (Fenwall‐Baxter, Deerfield, III.) were analyzed. Results: Mean platelet count was 237±49 × 103/mm3 (mean ± SD), and mean yield was 4.24±1.09 × 1011 platelets. Eighty‐five (12%) procedures generated less that 3 × 1011 platelets. Only thirty‐eight (5.4%) procedures yielded 6 × 1011 platelets, so that ‘split products’ could be obtained. Platelet yields were primarily related to the biologic contribution (baseline platelet count) of the donor. Procedure parameters selected for harvest, and the efficiency of the device also had a significant, but less important role in determining the final platelet yield. An increase in mean donor platelet count achieved with Mpl ligand therapy from 240,000 to 320,000/mm3 would reduce the cost from USD 378 to 267 for each apheresis product, since the fraction of split products would exceed 50% of apheresis procedures. Conclusion: Increasing the donor platelet count would have a significant economic impact on platelet apheresis programs, as well as important clinical consequences for the role of platelet apheresis products in future transfusion strategies.
Harvesting of Peripheral Blood Progenitor Cells with Different Programmes of Discontinuous Flow Systems
Authors: Moog R. , Uppenkamp M. , Böck H.P. , Müller N. , Brittinger G.
Published: December 2004

Background and Objectives:
From a technical point of view two problems arise during the collection of peripheral blood progenitor cells (PBPC): first, the volume of the apheresis product is often large, requiring volume reduction prior to cryopreservation. Second, the platelet (PLT) Ioss due to the harvesting of the buffy coat is an unwanted side effect. With respect to these problems, the present study was designed to compare programs for PBPC collection with discontinous‐flow cell separators. Materials and Methods: Three different protocols for PBPC harvesting were investigated in 32 patients with malignancies. In the first protocol, the blood cell separator Haemonetics MCS 3p was used. In the second protocol using the same machine, the opening and closure of the stem cell valve was modified in combination with a centri surge to reduce the PLT loss. The MCS+ device with another configuration of the valves was used in the third protocol. PBPC were mobilised by chemotherapy plus cytokine administration or application of growth factor alone. Blood counts and CD34 antigen‐expressing cells were determined before apheresis and in the PBPC product. Colony‐forming unit granulocyte/macrophage (CFU‐GM) were determined in the apheresis product. Results: 55 PBPC collections were carried out with a median end product volume of 105 ml. The median counts for CD34+ antigen‐expressing cells and CFU‐GM were 1.5 × 106 and 2.5 × 104/kg body weight, respectivel. PLT loss was significantly lower in protocol III. Conclusion: The study reported here revealed that PBPC could be easily collected in a reduced product volume by the intermittent‐flow cell separators. No additional centrifugation prior to cryopreservation was necessary to remove the plasma from the apheresis product. Patient's PLT loss was reduced by the centrisurge technique but this still has to be improved.
A Randomized Trial of Acute Normovolemic Hemodilution Compared to Preoperative Autologous Blood Donation in Total Knee Arthroplasty
Authors: Goodnough Lawrence Tim , Monk Terri G. , Despotis George J. , Merkel Kurt
Published: December 2004

The value of acute normovolemic hemodilution (ANH) compared to preoperative autologous blood donation (PAD) in elective surgery is controversial. We therefore conducted a prospective, randomized study to compare these techniques in patients undergoing total knee arthroplasty. ANH patient underwent up to 4 units phlebotomy or to target hematocrit level of 28% after induction of anesthesia. PAD patients were asked to donate 1 (unilateral) or 2 (bilateral, revisions) units before admission. Mean baseline hematocrit levels were not different between ANH and PAD patients (40.6±4.1 vs. 38.4±3.4, p = 0.09). Eight (73%) of 11 patients undergoing bilateral revision procedures received a total of 22 allogeneic blood units, whereas only 3 (14%) of 21 patients undergoing primary, unilateral procedures received a total of 3 allogeneic units (p = 0.002). We found no differences in allogeneic blood transfusions between ANH and PAD cohorts for all (n = 32) patients (1.0±1.2 vs. 0.6±1.4, p = 0.45), for unilateral knee (n = 21) replacement (0.25±0.46 vs. 0.08±0.28, p = 0.29), or for bilateral/revision (n = 11) procedures (1.9±1.3 vs. 2.5±1.9, p = 0.53). We conclude that each technique is equally effective in reducing allogeneic blood exposure. Patients undergoing revision or bilateral knee arthroplasties require adjunctive therapy to autologous blood procurement to further reduce allogeneic blood exposure.
Adverse Events in Platelet Apheresis Donors: A Multivariate Analysis in a Hospital‐Based Program
Authors: Despotis G.J. , Goodnough L.T. , Dynis M. , Baorto D. , Spitznagel E.
Published: December 2004

Objectives:
This study was designed to review the incidence of adverse events during nearly 20,000 apheresis procedures over a 4‐year period in a hospital‐based program. Methods: Data were obtained from a review of: (1) apheresis adverse event forms (2) hospital or emergency room medical records (3) the databank for donor and procedure‐related variables. Adverse events during or after the apheresis procedures were analyzed according to the following categories: (1) complications related to citrate toxicity; (2) hypotensive or vasovagal episodes; (3) complications or symptoms consistent with coronary ischemia; (4) complications related to percutaneous needle insertion, and (5) miscellaneous procedure‐related events or nonspecific symptoms. Serious adverse events were categorized as persistent or severe hemodynamic changes as well as other events that required further medical evalution. Results: Of 19,736 apheresis procedures, 159 (0.81%) were associated with adverse events. In 2,376 first‐time donations, 26 (1.09%) developed adverse events compared to 133 (0.77%) of 17,360 repeat procedures (p = 0.10). Seventy (0.35%) of 159 donation‐related adverse events involved hemodynamic or citrate‐related complications and 73(0.37%) involved venipuncture‐related complications, of which 2 required subsequent neurologic consultation. The remaining 23 (0.12%) adverse events involved procedure‐related, nonspecific complications. Forty‐seven (0.24%) of the 19,736 apheresis procedures were associated with serious adverse events (SAEs). Seven of these serious adverse events required admission to an emergency department, and 2 required admission to an emergency department, and 2 required hospitalization for further evaluation. Multivariate analysis revealed that apheresis machine model, donor gender and weight, the concomitant harvesting of plasma, the frequency of donation, and citrate‐related symptoms (e.g. paresthesias) were independently associated with severe hypotensive reactions. Conclusions: Apheresis procedures have a 150‐fold higher incidence of SAEs requiring hospitalization compared to whole blood donation. Identification of donors at risk for complications can facilitate modification of the apheresis procedure in order to reduce the likelihood of adverse events. Although our study did not demonstrate a cause‐effect relationship between platelet donation and the development of acute coronary syndromes; underlying cardiovascular disease was detected in 2 donors during or after the apheresis who were otherwise asymptomatic.
Press Review
Authors: M Ghio , MGP Contini , C Mazzei , S Brenci
Published: December 2004

Soluble HLA Class I, HLA Class II and Fas Ligand in Blood Components: A Possible Key to Explain the Immunomodulatory Effects of Allogeneic Blood Transfusion
Systems Contributing to the Assurance of Transfusion Safety in the United Kingdom
Authors: Williamson Lorna M.
Published: December 2004

In 1996, the United Kingdom launched a voluntary ‘haemovigilance’ system for confidential reporting of transfusion‐related deaths and major adverse events. The Serious Hazards of Transfusion (SHOT) initiative provided the first comprehensive overview of transfusion safety in the UK, with 12 fatalities reported in the first year. The most important finding was that of a total of 169 reports, 47% were ‘wrong blood to patient’ episodes, of which 16 were ABO‐incompatible. There were eight transfusion‐transmitted infections, three bacterial, four viral and one malarial. A number of other initiatives exist in the UK which also have importance in contributing to transfusion safety. This article reviews these other key contributors, allowing SHOT to be placed in context.
Collection of Heparinized Plasma by Plasmapheresis
Authors: van der Meer P.F. , Vrielink H. , Pietersz R.N.I. , Dekker W.J.A. , Reesink H.W.
Published: December 2004

Background and Objectives:
Heparinized plasma can be used for exchange transfusions in neonates and is usually collected by drawing whole blood using heparin as anticoagulant. The heparinized red blood cells and buffy coat cannot be used and are therefore discarded. To collect heparinized plasma more efficiently, a method was developed using an apheresis machine. Materials and Methods: With an MCS3p apheresis machine (Haemonetics), plasma was collected from volunteer donors as anticoagulant, heparin in saline (30,000 IU/I) was added in a 1:9 ratio. The activated partial thromboplastin time (APTT) of the donors was measured before and immediately after the procedure, and various parameters were determined in the collected plasma. Results: In 2 collection cycles, an average of 456±52 ml (mean ± SD; n =20) of heparinized plasma was collected, and 504±57 ml (n=2; donors with a high hemoglobin level) when 3 cycles were performed. The leukocyte and platelet contamination in the plasma (n =2) was 1.11±0.92×106 and 0.05±0.22×109 per unit, respectively, which conformed to national specifications. Sodium levels were normal, but due to dilution of the plasma with heparin solution, potassium and calcium levels were about 20% lower than the serum levels in the donors. The donor APTT values were slightly longer after the procedure than before, but remained all within normal values. Conclusion: For the collection of heparinized plasma, apheresis has the advantage that (1) high‐quality heparinized plasma can be harvested; (2) no blood components need to be discarded; (3) more plasma can be harvested with each donation, and (4) these procedures can be performed more often than whole blood donations.
Typing for the Human Lewis Blood Group System by Quantitative Fluorescence‐Activated Flow Cytometry: Large Differences in Antigen Presentation on Erythrocytes between A1, A2, B, O Phenotypes
Authors: Larson Göran , Svensson Lola , Hynsjö Lars , Elmgren Anders , Rydberg Lennart
Published: December 2004

Background:
Lewis phenotyping by hemagglutination is an unreliable routine method for Lewis antigen designation. Now genomic typing of the Lewis gene is available. Additionally, flow cytometry has been used for typing. We wanted to compare the results of Lewis typing in healthy individuals using the three methods. Materials and Methods: Ninety‐three randomly selected plasma donors were genotyped for inactivating Secretor (FUT2) G428A and Lewis (FUT3) T59G, T202C, C314T, G508A and T1067A point mutations: All Le(a+b−) individuals (nonsecretors) were homozygous for the FUT2 G428A mutation and all Le(a−b−) individuals had inactivating mutations on both FUT3 alleles. Fixed erythrocytes were analyzed by fluorescence‐activated flow cytometry and the results were compared with hemagglutionation and genotypic data. Antigen availability was expressed as median fluorescence intensity and as percentage positive cells with fluorescence intensities ≥ 102. Results: Using an anti‐Lea reagent a mean of 99% of erythrocytes from Le(a+b−) individuals and 1% of erythrocytes from Le(a−b−) or Le(a−b+) individuals were stained positive. Using an anti‐Leb reagent, a mean of 71% of erythrocytes from A1, 95% from B and 99% from O and A2 Le(a−b+) individuals and less than 10% of erythrocytes from Le(a−b−) or Le(a+b−) individuals were stained positive. After papain treatment 100% of the erythrocytes from A1 and A1B Le(a−b+) individuals stained positive without increase in background staining. The flow cytometric technique revealed large differences in staining intensities, within each ABO Le(a−b+) subgroup which was not directly correlated to plasma donation frequencies nor to Secretor or Lewis genotypes. Conclusion: Flow cytometry may prove valuable as a Lewis blood group typing technique but also as a research tool when investigating Lewis phenotypes of human erythrocytes.
Extracorporeal Plateletpheresis Induces the Interaction of Activated Platelets with White Blood Cells
Authors: Gutensohn K. , Alisch A. , Krueger W. , Kroeger N. , Kuehnl P.
Published: December 2004

Background and Objectives:
In this study we investigated whether platelet activation during apheresis results in the binding of platelets to white blood cells. Material and Methods: Analysis of platelet–leukocyte interaction was performed using multiparameter, three–color flow cytometry. Results: Over the duration of the procedure, there was an increase in the surface expression of CD62p (P–selectin) and CD63 (p<0.05), and also in the binding of platelets to monocytes (p<0.05), neutrophilic granulocytes (p<0.05) and to CD3+ cells (initially to a low degree; p<0.05). Platelet binding to CD19+ cells did not change significantly. Conclusion: This study demonstrates that platelets become activated during apheresis and that following this process, interaction with monocytes and neutrophilic granulocytes occurs.
Therapeutic Immunoadsorption Increases the Level of Circulating Soluble HLA Molecules
Authors: Poehlau D. , Kiltz U. , Rieks M. , Hoffmann V. , Rebmann V. , Schimrigk S. , Grosse–Wilde H. , Przuntek H.
Published: December 2004

Background and Objectives:
Immunoadsorption (IA) is an established procedure to remove Igs and immune complexes from peripheral blood. Since Igs reportedly bind to human leucocyte antigen (HLA) molecules, we were interested to know whether removal of Ig will also influence the plasma concentration of soluble HLA (sHLA). Patients and Methods: Nine patients suffering from severe autoimmune disease and undergoing 17 single courses of IA treatment were monitored for their sHLA class I (sHLA–I) and sHLA–DR plasma levels. Plasma was separated by a hollow–fiber–type separator. Plasma samples were taken before therapy, after 15 min of recirculation (without operating the adsorber), after 1 and 2 liters of plasma adsorption, and 24 and 48 h after the end of IA. Results: Before treatment the mean levels of sHLA–I and sHLA–DR were 0.37 (±0.06 SEM) and 0.32±0.05 μg/ml, respectively. After 2 liters of plasma filtration, an increase in sHLA–DR (0.80±0.10 μg/ml) was observed (p<0.001), whereas sHLA–I was only slightly affected (mean: 0.45±0.06 μg/ml). sHLA concentrations returned to initial levels after 24 h. Conclusion: The significant increase in sHLA–DR may contribute to the immunomodulatory effect of IA.
Photochemical Inactivation of Bacteria and HIV in Buffy‐Coat‐Derived Platelet Concentrates under Conditions That Preserve in vitro Platelet Function
Authors: Knutson Folke , Alfonso Ryan , Dupuis Kent , Mayaudon Véronique , Lin Lily , Corash Laurence , Högman Claes F.
Published: December 2004

Background and Objectives:
A photochemical process has been tested for the inactivation of viruses and bacteria in buffy‐coat derived platelet concentrates (BC PCs). Materials and Methods: BC PCs in 35% CPD plasma and 65% platelet‐additive solution (PAS III) were exposed to photochemical treatment (PCT) with 150 μM of the psoralen S‐59 and a 3 J/cm2 treatment with longwavelength ultraviolet light (UVA, 320–400 nm). Platelet function was evaluated following PCT using a panel of in vitro assays. Results: This PCT process was highly effective at inactivating gram‐positive bacteria (Staphylococcus epidermidis, Staphylococcus aureus, Enterococcus faecalis) and gram‐negative bacteria (Enterobacter aerogenes, Pseudomonas aeruginosa, Serratia marcescens). No viable bacteria were detected following PCT and 7 days of platelet storage while bacterial growth was detected in paired untreated control BC PCs. Complete inactivation of the gram‐positive Bacillus cereus was achieved only in one of two replicate experiments with BC PCs. PCT was also highly effective for inactivation of human immunodeficiency virus HIV‐1 in BC PCs inoculated with approximately 106 tissue culture infectious doses per milliliter (TCID5n/ml) of cell‐associated HIV‐1. Rapid inactivation was observed with increasing UVA doses: with 150 μM S‐59 and a 1 J/cm2 treatment of UVA, a reduction of 5:6±0.5 log TCID50/ml was achieved, and a reduction of >6.4 log TCID50/ml was achieved with 150 μM S‐59 and a 3 J/cm2 treatment of UVA. No physiologically relevant differences in platelet functions were found between the test and the control BC PCs during 7 days of storage. Conclusion: PCT with 150 μM S‐59 and a 3 J/cm2 UVA treatment does not adversely affect in vitro properties of BC PCs stored at 22°C for 7 days. The PCT process inactivated bacteria and HIV‐1 inoculated into the BC PCs. These results extend the earlier reported efficacy of PCT apheresis PCs to BC PCs.
An Extended Lymphocytotoxicity Test for Patients Treated with Lymphocytotoxic Antibodies
Authors: Moldenhauer A. , Kohlhaus R. , Salama A.
Published: December 2004

Background and Objectives:After organ transplantation, HLA antibodies (HLA‐Ab) stimulated by transplant or transfusion are usually indistinguishable from passive therapeutic lymphocytotoxic antibodies (LyAb), e.g. antilymphocyte globulin (ALG), antithymocyte globulin (ATG) and OKT3, by standard lymphocytotoxicity tests (LCTs). Herein, an extended technique capable of distinguishing between these two types of antibodies is described.Materials and Methods:Serum samples from 11 patients who received therapeutic antibodies were screened for HLA‐Ab by LCTs. The administered LyAb were murine OKT3 (n = 2), equine ALG (n = 3), rabbit ATG (n = 2), and combinations of ALG and OKT3 (n = 1), ATG and ALG (n = 1), and ATG and OKT3 (n = 2). To discriminate between therapeutically applied antibodies and active HLA alloimmunization, the sera were preincubated with species‐specific anti‐Fc lgG, thus blocking the therapeutic antibodies.Results:The sera of all patients treated showed strong positive lymphocytotoxic reactions [panel reactive antibodies (PRA) 50–100%]. After incubation with the corresponding Fc antibodies, in 9 of the 11 samples the amount of PRA decreased to the level present before transplantation (0–18%). In the remaining cases, new‐onset alloimmunization was detected.Conclusion:This extended LCT allows the screening of serum samples for active HLA‐Ab even in the presence of therapeutic LyAb.
Confirmation of the Assignment of the Dombrock Blood Group Locus (DO) to Chromosome 12p: Narrowing the Boundaries to 12p 12.3‐p13.2
Authors: Mauthe Jill , Coghlan Gail , Zelinski Teresa
Published: December 2004

Background and Objectives
: Antigens belonging to the Dombrock blood group system have been well characterized serologically. Despite being reasonably polymorphic, chromosomal localization of the gene controlling Dombrock expression (DO) has proved difficult. In the past, DO had been provisionally assigned to chromosomes 1 and 4; more recently, DO was provisionally assigned to chromosome 12. The current study was undertaken in an attempt to confirm or refute this latest assignment. Materials and Methods: DNA from a series of families segregating for DO was amplified by polymerase chain reaction and analyzed for trinucleotide or tetranucleotide repeat polymorphisms of four anonymous DNA markers (D12S1042, D12S373, D12S391, D12S372) and of the von Willebrand factor gene (VWF). Lods for or against linkage were determined between DO and each marker. Results: DO is linked to all five markers. The closest linkage was established between DO and D12S373, and DO and D12S391, with peak lods for combined paternal and maternal meioses of 7.16 at peak recombination fraction (

) = 0.08 and 7.69 at

= 0.08, respectively. Conclusions: We have confirmed the assignment of the Dombrock blood group locus to chromosome 12p and have refined its regional localization to 12p 12.3–p 13.2.
Low Leukocyte Contamination without Filtration by Preparation of Platelet Concentrates in Cylindrical Bags with the Buffy‐Coat Method
Authors: Kostelijk E.H. , Loos J.A. , Veldman H.A. , de Korte D.
Published: December 2004

Background and Objectives:
The buffy‐coat (BC) method for platelet concentrate (PC) preparation was modified in order to obtain leukodepleted PCs from single BCs without filtration. Materials and Methods: BCs were centrifuged in cylindrical BC bags and the optimal centrifugation conditions and optimal hematocrit were determined. Results: With optimal conditions, a tenfold lower leukocyte contamination was obtained compared with the conventionally shaped, wide BC bag (0.3 ± 0.19 versus 3.0 ± 1.71 × 106 leukocytes per unit; 85‐ml BCs). The platelet yield obtained with the cylindrical bag did not differ significantly from the yield obtained with the conventional bag (56 ± 16.4 versus 61 ± 15 × 109 platelets per PC). Furthermore, when PCs were prepared from 100‐ml BCs in cylindrical bags, a leukocyte contamination of 0.2 ± 0.11 × 106 and a platelet content of 6 ± 13.5 × 109 per PC were obtained. Conclusion: The use of cylindrical BC bags reduced the leukocyte contamination in PCs to a level required for leuko‐depletion without affecting platelet recovery.
Automated Collection of Peripheral Blood Stem Cells with the COBE Spectra for Autotransplantation
Authors: Heuft H.G. , Dubiel M. , Kingreen D. , Oertel J. , De Reys S. , Rick O. , Serke S. , Schwella N.
Published: December 2004

Background:
A convenient, effective and safe peripheral blood stem cell (PBSC) apheresis procedure is desirable to cope with the increasing requirements for PBSC collections. We performed PBSC harvesting with the novel COBE Spectra Auto PBSC™ system using the default software configuration recommended by the manufacturer. We analyzed collection parameters and clinical efficiency of harvested autografts following high‐dose chemotherapy (HDCT). Patients and Methods: Eighty‐one patients underwent 102 harvests after standard chemotherapy plus filgrastim (5–10 ug/kg/day) to obtain a target of ≥2.5 × 106 CD34+ cells/kg for autologous blood stem cell transplantation. Conventional‐volume leukaphereses (median: 11 leters) were performed using the manufacturer's standard software default regarding inlet flow, harvest/chase volume (3/7 ml) and number of collection cycles. The ratio of ACD‐A to whole blood was initially set at 1:12 (56 collections), later at 1:10 (46 aphereses). Results: With respect to preapheresis counts of 93 (9–876) CD34+ cells/μl, 69 patients (85.2%) achieved ≥2.5 × 106 CD34+ cells/kg by the first apheresis. PBSC products contained medians of 5.0 × 106 (0.7–77.3) CD34+cells/kg and 13.8 × 104 (2.3–105.0) CFU‐GM/kg. A preapheresis count of ≥40 CD34+ cells/μl predicted a single‐apheresis yield of ≥2.5 × 106 CD34+ cells/kg. Apheresis products showed a high mononuclear cell (MNC) purity of ≥89%. The median overal collection efficiency of CD34+ cells (CD34‐CE) was 42.6% (12.2–87.4). The CD34‐CE decreased significantly with increasing numbers of circulating CD34+ cells: 52.5% at CD34+ cells <40/μl versus 41.0% at CD34+ cells ≥40/μl (p ≤ 0.002). Coagulation problems occurred in 3 of 56 procedures at an ACD‐A to whole blood ratio of 1:12, but in no case at 1:10. Patients who underwent HDCT and autologous blood stem cell transplantation (n=40) engrafted in a median time of 9 (8–13) days for absolute neutrophil counts ≥ 0.5 × 103/μl, 10 (8–13) days for WBC > 1.0 × 103/μl and 11 (8–17) days for platelets > 20 × 103/μl. Conclusions: As a result of efficient PBSC mobilization, a single conventional‐volume leukapheresis with the COBE Spectra Auto PBSC system resulted in hematopoietic autografts with ≥ 2.5 × 106 CD34+ cells/kg in 85% of patients. Following the standard PBSC apheresis recommendations of the manufacturer, the Auto PBSC system assures PBSC products with a high MNC purity and a moderate CD34‐CE that declines significantly at increasing levels of circulating CD34+ cells. Leukaphereses performed at an ACD‐A to whole blood ratio of 1:10 should run without coagulation problems.
An Experiment with Glycerol‐Frozen Red Blood Cells Stored at ‐80°C for up to 37 years
Authors: Valeri C.R. , Ragno G. , Pivacek L.E. , Cassidy G.P. , Srey R. , Hansson‐Wicher M. , Leavy M.E.
Published: December 2004

Abstract
Background and Objectives:
Red cells frozen using 40% W/V glycerol are currently FDA approved for frozen storage at ‐80°C for up to 10 years. Materials and Methods: Red cells frozen with 40% W/V glycerol and stored at ‐80°C for up to 37 years were thawed, deglycerolized, and stored at 4°C for 24 h. Results: Red cells frozen for up to 37 years had mean freeze‐thaw‐wash recovery values of 75%, less than 1% hemolysis, and normal ATP, 2,3‐DPG and P50 levels, and 60% of normal RBC K+ levels. Conclusions: Red cells frozen with 40% W/V glycerol can be stored at ‐80°C for up to 37 years with acceptable in vitro results.
Press Review
Authors: Engelfriet C.P. , Vadhan‐Raj S.
Published: December 2004

Clinical Experience with Recombinant Human Thrombopoietin in Chemotherapy‐Induced Thrombocytopenia
A UK Survey of Virological Testing of Cadaver Tissue Donors
Authors: Stanworth S.J. , Warwick R.M. , Ferguson M. , Barbara J.A. ,   Microbiology Working Group of the NIBSC steering group on Tissue/Cell Banking and Engineering
Published: December 2004

Objectives:To analyse information about virological testing of cadaveric tissue donors, including the kits used and the rates of test reactivity.Materials and Methods:Retrospective data were collected using a standardised questionnaire sent to 16 tissue banks in the UK. The rates of repeat reactive screen tests and confirmed positive results for the markers HBsAg, anti‐HCV, anti‐HIV were analysed in 1,833 cadaver tissue donors tested at 12 tissue banks over one year up to March 1998.Results:There was a wide range of kits in use for virological screen testing of cadaver donors. The rates of repeat reactivity in the screen tests varied from 0 to 42%.Conclusion:The findings have implications for policies based on discard of tissues from donors with repeat reactive results, and raise important safety issues with regards to cadaveric virological testing, as the test systems in use have not been validated for cadaver blood samples.
Evaluation of depth filtration to remove prion challenge from an immune globulin preparation
Authors: Van Holten R. W. , Autenrieth S. M.
Published: December 2004

Background and Objectives Plasma‐derived therapeutic proteins have the potential to contain transmissible spongiform encephalopathy (TSE) infectivity. This study evaluated the effectiveness and characterized the mechanism of abnormal prion protein removal during a depth‐filtration step used in the manufacture of an immunoglobulin preparation.

Materials and Methods Scrapie brain homogenate was treated with lysolecithin, sonicated and sequentially filtered through 0·45‐, 0·22‐ and 0·1‐µm membrane filters. The scrapie brain homogenate was then added (at a 1 : 51 dilution) to the Supernatant III fraction used in the manufacture of Rho(D) immune globulin (human). The spiked immunoglobulin preparation was then filtered through a depth filter under the same conditions used in full‐scale production. After filtration, the depth filter was washed with hypertonic NaCl solutions to elute the abnormal prion protein (PrPSc) from the filter. A Western blot assay for PrPSc was used to quantify removal from the filtrate and recovery from the filter washes. A second run was performed whereby the PrPSc‐spiked Supernatant III was filtered through a 0·22‐µm membrane filter prior to depth filtration. A third run evaluated depth filtration of PrPSc in Tris‐buffered saline (TBS).

Results The depth filter removed greater than four logs of PrPSc from the Supernatant III filtrate. A significant portion of the PrPSc could be recovered from the depth filter by elution with high‐molarity NaCl solutions. Prefiltration (through a 0·22‐µm membrane filter) of the spiked Supernatant III prior to depth filtration removed all detectable PrPSc. Depth filtration removed less than one log of PrPSc from TBS.

Conclusions Depth filtration appears to remove PrPSc from the immunoglobulin preparation by mechanical straining rather than by adsorption to the filter matrix. The immunoglobulin preparation caused the PrPSc to aggregate from particles < 0·1 µm in size to particles of > 0·22 µm, probably as a result of the presence of methanol in the preparation. The depth filter failed to remove PrPSc from a purely aqueous environment.
Reduction of the diagnostic window in three cases of human immunodeficiency‐1 subtype E primary infection with fourth‐generation HIV screening assays
Authors: Weber B. , Thorstensson R. , Tanprasert S. , Schmitt U. , Melchior W.
Published: December 2004

Background and Objectives The influence of genetic variability on the sensitivity of serological diagnosis of human immunodeficiency virus (HIV) infection has, to date, been poorly investigated. The aim of the present study was to assess whether fourth‐generation assays for the combined detection of HIV antigen and antibodies to HIV (anti‐HIV) permit a reduction of the diagnostic window in comparison to third‐generation antibody enzyme immunoassays (EIAs), which so far have shown a poor sensitivity for detection of HIV‐1 non‐subtype B primary infections.

Materials and Methods Three patients with primary HIV‐1 subtype E (CRF01‐AE) infection were tested with different third‐ and fourth‐generation assays, stand‐alone HIV antigen (Ag) EIAs and reverse transcription–polymerase chain reaction (RT–PCR). Additionally, virus lysates from HIV‐1 Group M and O and HIV‐2, at concentrations of p24 Ag close to the detection limit of licensed HIV Ag EIAs, were investigated with fourth‐generation EIAs and HIV Ag EIAs.

Results In the first blood donor, the most sensitive fourth‐generation assay detected HIV‐1 infection 11 days earlier than five of the eight third‐generation antibody assays. Fourth‐generation EIAs, with a high sensitivity for HIV antigen, detected HIV‐1 subtype E infection simultaneously or 4 days later than HIV‐1 RT–PCR on pooled samples. Low concentrations of virus lysates of different HIV‐1 subtypes A–H and group O, tested positive with fourth‐generation EIAs, with a high sensitivity of the antigen‐detection module.

Conclusions Fourth‐generation EIAs, especially those with a high sensitivity for HIV‐1 p24 antigen, reduce the diagnostic window for primary HIV‐1 subtype E infection in comparison with third‐generation antibody‐screening assays. These preliminary data from seroconversions and virus lysates indicate that the genetic diversity of HIV‐1 does not represent a major challenge for the most sensitive EIAs of this new assay generation.
Early postnatal development of central corneal thickness in dogs
Authors: Montiani‐Ferreira Fabiano , Petersen‐Jones Simon , Cassotis Nicholas , Ramsey David T. , Gearhart Patricia , Cardoso Fernando
Published: December 2004

ObjectiveTo investigate the changes in corneal thickness that occur during maturation of the canine eye over the first months of life.

Animals studiedDogs of two different breeds with ages ranging from 14 days to 42 weeks of age.

ProceduresThe central corneal thickness was measured by ultrasonic pachymetry every week for the first month after eyelid opening (around 14 days) and then every month until 42 weeks of age. Segmented regression was applied to capture the two phases observed in the central corneal thickness plotted against age. Breed, eye and gender were also included in the model.

ResultsMean central corneal thickness (CCT) values initially decreased following eyelid opening, with the lowest point being reached at around 6 weeks of age. Then CCT gradually increased as the dogs matured. Differences between left and right eye were not significant. Breed and gender effects were significant factors in the statistical model.

ConclusionsFollowing eyelid opening there is an initial decrease in corneal thickness until approximately 6 weeks of age, which presumably mirrors maturation of corneal endothelial cell function. After 6 weeks of age the CCT increases with age until approximately 30 weeks of age after which there was only a gradual increase over the remainder of the study period. A similar pattern of changes in corneal thickness in humans has been previously recorded.
The effects of topical ocular application of 0.25% demecarium bromide on serum acetylcholinesterase levels in normal dogs
Authors: Ward D. A. , Abney K. , Oliver J. W.
Published: December 2004

ObjectiveTo assess the effects of topical ocular application of 0.25% demecarium bromide on serum acetylcholinesterase (AChE) levels in normal dogs.

AnimalsNine adult mixed breed dogs weighing between 18 and 27 kg.

ProceduresFifty µL of 0.25% demecarium bromide were applied to one eye of each dog every 8 h for 6 days. Blood was analyzed for AChE levels prior to commencement of eye drops, and at 45 min, 1 h 45 min, 4 h 45 min, 1 day, 3 days, and 7 days following commencement of eye drops using a 5,5′‐dithiobis‐(2‐nitrobenzoic acid) (DTNB) reaction.

ResultsAcetylcholinesterase levels declined over the first 24 h following commencement of demecarium administration in most dogs. This decline was highly variable and was statistically significant by 24 h. In some individuals AChE levels were suppressed to levels approaching clinical toxicity. By day 3 AChE levels had risen to levels above baseline in most dogs.

ConclusionsTopical ocular application of demecarium causes transient suppression of systemic acetylcholinesterase levels in most dogs. Acetylcholinesterase levels generally do not fall to toxic levels, but may do so in certain individuals. Demecarium bromide eye drops generally do not cause AChE toxicity, but dogs receiving such therapy should be monitored for signs of AChE toxicity, and concomitant use of other AChE inhibitors should be avoided.
Seasonal effects on the aerobic bacterial and fungal conjunctival flora of normal thoroughbred brood mares in Florida
Authors: Andrew Stacy E. , Nguyen An , Jones Galin L. , Brooks Dennis E.
Published: December 2004

ObjectiveTo evaluate seasonal effects on the presence or absence of fungal and aerobic bacterial flora of the conjunctival fornix of normal Florida Thoroughbred horses.

Sample populationBoth eyes of 100 horses.

ProcedureHorses with normal anterior segment ophthalmic examinations from three farms in north central Florida were included. Each animal had the ventral conjunctival fornix of each eye swabbed with sterile culturettes. Samples were taken in October, January, April, and July (1999–2000). Aerobic and fungal cultures were plated. Bacterial cultures were reviewed at 24 and 48 h. Fungal cultures were reviewed weekly for 4 weeks. Logistic regression analysis with season as a factor and age of the horse as a covariate was performed. Statistical significance was set atP< 0.01.

ResultsHorses ranged from 3 to 24 years of age, with a median age of 9 years. Twenty‐four genera of bacteria and 35 genera of fungi were recovered.Corynebacteriumsp.,Staphylococcussp.,Bacillussp. andMoraxellasp. were the bacteria most frequently isolated. Mold species, dematiaceous mold species,Chrysosporiumsp.,Cladosporiumsp., andAspergillussp. were the most frequently recovered fungi. Season did not have a significant effect on the presence of microorganisms isolated for individual horses adjusted for age. Younger horses had an increased incidence of gram‐negative rods and fungal isolates. The number of bacteria and fungi isolated are not uniform across seasons.

ConclusionThere were no significant differences between the number or type of organisms cultured during the sampling seasons in normal Florida horses. A large range of normal bacterial and fungal flora were isolated from these horses. The number of bacteria and fungi isolated are not uniform across seasons. The likelihood of detecting an organism depends on the horses’ age.
Fungal flora of normal eyes of healthy horses from the State of Rio de Janeiro, Brazil
Authors: Rosa Maurílio , Cardozo Liane Maria , Da Silva Pereira Jorge , Brooks Dennis E. , Martins Ana Lucia B. , Florido Penha Sueli Silva , Stussi Jussara Schwind Pedroso
Published: December 2004

The conjunctival fungal flora of 32 adult horses with normal eyes (n = 64) from the State of Rio de Janeiro in Brazil was identified in the fall of 2000 using horses of different breeds, both genders and aged 5–19 years old. The culture samples were taken from the conjunctival sac of both eyes with a sterile cotton swab wetted with saline solution, seeded in Sabouraud's dextrose agar with chloramphenicol, and incubated for 5 days at an average temperature of 25 °C. The number of fungal colonies per eye varied between 0 and 250 colony forming units (CFUs). There were often differences in colony types between eyes of the same animal. Filamentous fungi of genera were isolated and identified in the following proportion of the total genera of fungal colonies isolated: Aspergillus (32.2%), Penicillium (25.8%), Scopulariopsis (15.9%), Trichoderma (11.2%), Cladosporium (5.6%), Mucor (2.1%), Syncephalastrum (2.1%), Eurotium (1.7%), Geotrichum (0.9%), Rhizopus (0.9%), Gliomastix (0.4%), Fusarium (0.4%), Staphylotrichum (0.4%) and Verticillium (0.4%). Yeast genera represented 9% of the total isolates. Over half the horses had at least one normal eye with either Aspergillus, Penicillium, Trichoderma or Scopulariopsis isolated, which is a departure from other studies of the normal horse eye.
Changes in intraocular pressure and pupil size following intramuscular administration of hydromorphone hydrochloride and acepromazine in clinically normal dogs
Authors: Stephan Deborah D. , Vestre W. A. , Stiles Jean , Krohne Sheryl
Published: December 2004

ObjectiveTo investigate the effects of intramuscularly administered hydromorphone hydrochloride and acepromazine on intraocular pressure (IOP) and pupil size (PS).

Animals studiedSeventeen dogs free of clinically relevant ocular abnormalities.

ProcedureMeasurements of IOP and PS were obtained and the dogs were injected intramuscularly with hydromorphone (0.04–0.08 mg/kg) and acepromazine (0.04 mg/kg). Measurements of IOP and PS were repeated 10 min and 25 min later.

ResultsThough a decreasing trend in IOP values was demonstrated, no significant difference was noted in IOP from the initial examination to examination following intramuscular administration of hydromorphone and acepromazine. Significant miosis was present in 16 of 17 dogs at 10 min and 25 min following administration of hydromorphone and acepromazine.

ConclusionHydromorphone (0.04–0.08 mg/kg) and acepromazine (0.04 mg/kg) cause significant miosis in dogs at 10 and 25 min following intramuscular administration.
Young Children's Evaluations of the Ingroup and of Outgroups: A Multi‐National Study
Authors: Bennett Mark , Barrett Martyn , Karakozov Rauf , Kipiani Giorgi , Lyons Evanthia , Pavlenko Valentyna , Riazanova Tatiana
Published: December 2004

In investigating intergroup attitudes, previous research in developmental psychology has frequently confounded ingroup favouritism and outgroup derogation. The present study, using unconfounded measures, examines the possibility that ingroup favouritism and outgroup derogation are distinct phenomena. Six‐year‐old children (n=594) from five, culturally diverse nations were asked to make various evaluations of the national ingroup and of four national outgroups. The data indicate that although there is overwhelming evidence that young children favour the ingroup over other groups, outgroup derogation is limited in extent and appears to reproduce attitudes held by adult members of the particular nations investigated.
Non-Ignorable Missing Covariate Data in Survival Analysis: A Case-Study of an International Breast Cancer Study Group Trial
Authors: Herring Amy H. , Ibrahim Joseph G. , Lipsitz Stuart R.
Published: December 2004

Non-ignorable missing data, a serious problem in both clinical trials and observational studies, can lead to biased inferences. Quality-of-life measures have become increasingly popular in clinical trials. However, these measures are often incompletely observed, and investigators may suspect that missing quality-of-life data are likely to be non-ignorable. Although several recent references have addressed missing covariates in survival analysis, they all required the assumption that missingness is at random or that all covariates are discrete. We present a method for estimating the parameters in the Cox proportional hazards model when missing covariates may be non-ignorable and continuous or discrete. Our method is useful in reducing the bias and improving efficiency in the presence of missing data. The methodology clearly specifies assumptions about the missing data mechanism and, through sensitivity analysis, helps investigators to understand the potential effect of missing data on study results.
A Bayesian Method for Estimating the Accuracy of Recalled Depression
Authors: Rutter Carolyn M. , Simon Gregory
Published: December 2004

We develop a Bayesian method that allows us to compare weekly depression states recalled for a 3-month period to cross-sectionally assessed measurements of current depression assessed during randomly timed phone interviews. Using these data, we examine the accuracy of recalled depression by linking a spline model for recalled depression and a logistic model for current depression. The logistic model includes the model-based probability of depression based on recall as a covariate and covariates potentially related to the accuracy of recall. The model that we propose allows variability in both measures and can be modified to examine general relationships between longitudinal and cross-sectional measurements.
A Litter-Based Approach to Risk Assessment in Developmental Toxicity Studies via a Power Family of Completely Monotone Functions
Authors: Kuk Anthony Y. C.
Published: December 2004

A new class of distributions for exchangeable binary data is proposed that originates from modelling the joint success probabilities of all orders by a power family of completely monotone functions. The distribution proposed allows flexible modelling of the dose–response relationship for both the marginal response probability and the pairwise odds ratio and is especially well suited for a litter-based approach to risk assessment. Specifically, the risk of at least one adverse response within a litter takes on a simple form under the distribution proposed and can be reduced further to a generalized linear model if a complementary log–log-link function is used. Existing distributions such as the beta–binomial or folded logistic functions have a tendency to assign too much probability to zero, leading to an underestimation of the risk that at least one foetus is affected and an overestimation of the safe dose. The distribution proposed does not suffer from this problem. With the aid of symbolic differentiation, the distribution proposed can be fitted easily and quickly via the method of scoring. The usefulness of the class of distributions proposed and its superiority over existing distributions are demonstrated in a series of examples involving developmental toxicology and teratology data.
Telephone use among noninstitutionalized persons with dementia living alone: Mapping out difficulties and response strategies
Authors: Nygård Louise , Starkhammar Sofia
Published: December 2004

The aim of this study was to map out and describe difficulties and response strategies in telephone use among elderly, noninstitutionalized persons with dementia living alone. To obtain explorative data, interviews and observations in the homes were undertaken with 10 participants diagnosed with dementia. The participants were observed when showing their telephones, calling a well‐known number, responding to a request previously sent by letter to make a telephone call, and finding a number in the telephone directories. The data were analysed using a comparative approach, and resulted in descriptive categories. The participants' difficulties were categorized as difficulty in ‘knowing what’, ‘knowing where’, ‘knowing how’ and overcoming motor, perceptual, verbal and environmental obstacles. In responding to these, they used a variety of environmentally related strategies such as using perception or habits and habitual places, verbalizing aloud, seeking help from others, and adjusting the physical environment. They also used a few cognitively related strategies such as repeating and stopping and reflecting. Overall, difficulties were frequent and common, and the effectiveness of the strategies was questionable. The results indicate that it might be unrealistic to assume that elderly persons with symptoms of dementia who live alone are able to satisfactorily use the telephone for safety, communication and participation in society.