bluuebunny/crossref_metadata_2025_split huggingface dataset:
YS05 In Vitro Evaluation of Platelet Concentrates Prepared Using the Automated Gambro OrbiSac System
Authors: Wiltshire M. , Bashir S. , Beard M. , Cookson P. , Garwood M. , Hancock V. , Pergande C. , Smith K. , Turner C. , Thomas S. , Cardigan R. 
Published: September 2006

Introduction  The Gambro OrbiSac device is designed to automate the preparation of platelet concentrates (PC) from buffy coats (BC). The aim of this study was to evaluate the in vitro function of PC produced using the OrbiSac system and stored in plasma.Study design  Ten PC were produced by automated pooling of four ABO matched BC and a single unit of plasma. The ring‐shaped pooling bag was centrifuged and the platelet rich supernatant automatically expressed, through a leucodepletion filter, into a Gambro ELP storage bag. All PC were stored at 22°C with agitation and sampled on days 1, 5, 7 and 9. Samples were evaluated for platelet count and standard markers of platelet quality.

Parameter
OrbiSac PC
Specification (Manual PC)

Volume (mL)
366 (327–396)
150–350 mL (Local)

Platelet Content (×109 U−1)
297 (251–419)
>240 (UK)

Median (range) n = 10.

The assay results were compared to platelets produced by manual methods and stored in a Pall ELX bag in a previous study.

Parameter
OrbiSac PC
Manual PC

Day 1
Day 5
Day 7
Day 9
Day 1
Day 5
Day 7
Day 9

pH (22 °C)
7.23
7.45
7.38
7.29
7.13
7.36
7.32
7.24

pO2 (kPa)
9.1
7.1
7.2
8.6
6.3
5.4
5.8
6.2

pCO2 (kPa)
4.5
1.8
1.7
1.7
5.9
2.6
2.5
2.4

HSR (%)
61.8†
54.3†
55.6
58.1
76.3
71.4
64.5
70.6

% +ve CD62P
25.5
21.9
28.9
36.2
22.1
16.6
31.3
43.1

% +ve CD63
12.9
18.5
15.9
16.9
9.4
11.3
14.2
17.2

Median n = 10.
†
P < 0.001 versus manual (T‐test).

Results  Hypotonic shock response (HSR) values were found to be significantly lower in this study when compared to manually produced PC. This was due to increased red cell contamination in these units. HSR was further investigated during a larger scale trial and all units (n = 25) showed improved day 1 values (78%, range: 61–93%). Values from these additional PC were comparable with those from manually produced PC.Conclusion  PC produced with the OrbiSac system achieved the local specification for platelet count. However, these units were higher in volume than those produced by manual methods, with 70% of units exceeding the locally specified maximum volume of 350 mL. This problem could be addressed during later trials. OrbiSac PC have acceptable pH, blood gases, HSR levels and platelet activation markers and results are broadly comparable to data generated in a previous study of manually produced PC. These results suggest that platelets produced with the Gambro OrbiSac system are comparable to those produced using the current manual methods.
SI35 Blood Group Antigen Typing Using DNA Microarray BeadChipsTM–First UK Report
Authors: Armstrong‐Fisher S. , Varzi A. M. , Urbaniak S. J. , Cavanagh G. 
Published: September 2006

Background  The determination of human blood group phenotypes has routinely involved red cell agglutination techniques. The genes for all but one of the 29 human blood group systems have now been cloned and the molecular bases of all clinically significant blood group polymorphisms are known. Consequently these DNA sequences can be used to predict blood group phenotypes. Molecular typing as an adjunct to serology offers improved accuracy and can save both rare reagents and laboratory time while screening for compatible donor units. This study determined the feasibility of analysis of single nucleotide polymorphisms (SNPs) associated with a variety of blood group antigens using a DNA microarray system with a beadchip technology platform.Method  SNPs were tested by elongation‐mediated analysis of polymorphisms (eMAPTM) using allele‐specific oligonucleotides with variable 3’‐terminal sequences attached to colour‐encoded beads. The beads are assembled into arrays on semiconductor chips (BeadChipTM), available from Bioarray Solutions. Elongation products for SNPs or small deletions/insertions were simultaneously detected by instant imaging of fluorescence signals from the entire array. Seventeen polymorphisms associated with Human Erythrocyte Antigens and one mutation associated with hemoglobinopathies (CO, DI, DO, FY, JK, KEL, LU, LW, MNS, SC and HgbS) were included in the HEA BeadChipTM Kit.Results  The system was validated using a reference DNA panel of known phenotype (n = 30) and genotype, established by conventional DNA‐PCR SSP assays. Replicate testing (n = 6 reps for eight samples) gave 100% consistent data demonstrating the Bioarray system to be both reliable and reproducible. The initial concordance rate with serology was 99.2%.Conclusion  The BeadChip System represents the first integrated flexible array solution for high complexity transfusion testing, accommodating ‘multiplex’ analysis of polymorphisms. This high throughput, microarray system offers the potential to determine extended genotypes of patients and maintain an extended‐genotype, national panel of donors to provide compatible blood for sensitised patients. The initial high concordance rate and the rarity of CE marked phenotyping reagents justifies its further development.
SI37 Two Novel D Genes of the Rh Blood Group System Producing D Variant Phenotypes
Authors: Etheridge W. , Tilley L. , Poole J. , Daniels G. 
Published: September 2006

Introduction  The D antigen is the most clinically significant of the Rh blood group system. Many variants of D exist, which are often classified as weak or partial D. D variants usually result either from nucleotide changes encoding amino acid substitutions or from the replacement of sections of RHD by the equivalent region of RHCE. Amino acid substitutions within the cytoplasmic or transmembrane domains of the RhD protein often produce weak D phenotypes with quantitative changes to D antigen expression, whereas substitutions in the extracellular loops of the RhD protein often result in qualitative changes associated with partial D phenotypes. This dichotomy, however, is not definitive.Samples and Methods  Molecular analysis was performed on 14 samples referred for investigation of RhD status. Genomic DNA was extracted from whole blood. Exons 1–10 of RHD were sequenced for all samples and compared to reference sequences. Serological data were obtained where possible.Results  Nucleotide mutations were found in all samples. Most could be assigned to known D variant types, but two novel mutations were detected. In four out of five samples identified as a possible DOL D variant by serological testing, an extra mutation was found. DOL variants have T>C509 in exon 4 (M170T) and T>G667 in exon 5 (F223V). In addition a C>G1132 mutation in exon 8, encoding L378V in the twelfth transmembrane domain, was present. This variant of DOL has subsequently been reported as DOL‐2 (http://www.uni‐ulm.de/~wflegel/RH). The second novel mutation, A>G485 in exon 3 encoding N162S, was found in one person with alloanti‐D. The substitution is predicted to be in extracellular loop three of the RhD protein.Conclusions  Two novel D variants were identified. One resembles DOL and is called DOL‐2. The other, which we propose to name DNS, was found in a person with anti‐D.
P17 Bedside Practice of Blood Transfusion ‐ Where is the Evidence?
Authors: Brunskill S. , Doree C. , Blest A. , Murdock J. , Roberts M. , Watson D. 
Published: September 2006

Background  There have been no systematic reviews of the routine ‘bedside practice’ of blood transfusion. There are national guidelines, but local practice varies. Nursing staff are increasingly requesting details of the evidence that underpins the guidelines. Transfusion practitioners from the four UK blood transfusion services working with the NBS Systematic Review Initiative have systematically reviewed the evidence for practice in four key areas of bedside blood transfusion.Methods  Relevant evidence was identified through comprehensive searches of electronic databases: Medline; Embase; Cinahl; The Cochrane Library; websites of international blood transfusion societies and the websites of the US Centres for Disease Control and Prevention, English National Patients Safety Agency and the NHS Serious Hazards of Transfusion Scheme. Reference lists of relevant papers were scanned and contact was made with transfusion medicine clinicians and scientists to identify additional evidence. All identified papers were assessed for project eligibility: eligible papers were quality assessed and data extracted by two members of the project team, working independently of each other.Results  The searches yielded approx. 6000 hits. Relevant evidence was identified for one area: co‐administration of drugs with blood (approx. 50 papers). Indirectly relevant evidence was identified for the maximum time that a unit of blood can be safely left out of refridgeration (approx. six papers). No relevant published information was identified for the frequency of change of blood administration or for the relative safety of 1 versus 2 person checking blood prior to infusion.Conclusions  For three of the four areas despite minimal restrictions on study design there was little relevant published evidence to underpin practice. Where evidence does exist, it is not necessarily directly related to bedside blood transfusion practice or is ‘out‐of‐date’. The principal recommendation from this project is the need to undertake directly relevant primary research, including assessing the bacterial contamination of blood during the transfusion process and a national randomised controlled trial of the safety of 1 versus 2 person checking.
P02 Inherent Variation in Donor Platelet Reactivity Contributes to the Quality of Apheresis Platelets
Authors: Garner S. F. , Jones C. I. , Stephens J. , Carr P. , Walton J. , Bernard A. , Angenent W. , Ouwehand W. H. , Goodall A. H. 
Published: September 2006

Introduction  Provision of platelets is a major commitment for Blood Services. Iterative cycles of improvements in platelet procurement and storage have lead to a better quality of concentrates, but even the latest generation of apheresis machines may produce activated platelets. Less attention has been paid in determining the contribution of an individual donor's inherent platelet reactivity on the activation state of apheresis platelets. We investigated this donor related variable using a panel of donors from the BLOODOMICS project whose platelet responses to activation through the ADP and collagen pathways were known to represent the widest range of the normal responses. For the current study three groups of donors with responses that were characterised as high (n = 16), low (n = 15) or in the mid‐range (n = 16), were recalled and the activation state of their circulating platelets and donated apheresis platelets were measured.Method  To assess the activation state of platelets in the circulation, prior to donation samples were taken using a 21G butterfly needle, without a tourniquet. Aliquots of platelets from the apheresis donation were taken immediately at the end of the procedure. The level of platelet‐bound fibrinogen, a sensitive marker for early‐stage activation was measured using a whole‐blood flow cytometric assay.Results  Prior to donation resting bound fibrinogen levels in all subjects ranged from 1.6–11.1% (mean 3.6%), and there was a significant association between this index of baseline platelet activation and the donor's responder status (P = 0.009). The percentage of platelets with bound fibrinogen in the apheresis concentrates ranged from 0.6–84.3% (mean 14.7%) and were generally higher than resting levels. Increased apheresis‐induced activation was associated with the responder status of the donor (P = 0.005). In particular, the 4 platelet concentrates with the highest levels of bound fibrinogen (>50%) were all from high responder donors.Conclusion  Overall it is not solely the process of donation that affects platelet quality. Platelets that show higher responses to agonists are particularly susceptible to activation during collection by apheresis. We postulate that the responses to agonists are largely genetically controlled, and are attempting to identify the relationship between variation in platelet‐gene sequence and platelet function.
P24 Use of Allogenic Blood in Total Hip and Knee Arthroplasty: Need for a National Protocol
Authors: Mangwani J. , Gardiner M. , Williams W. W. 
Published: September 2006

Introduction  Patients undergoing Total hip replacement (THR) and total knee replacement (TKR) surgery are usually elderly and have other co‐morbidities. This combined with the nature of the surgery ensures that a number will have postoperative blood transfusion. Blood transfusion carries a number of well document risks for the patient and is increasingly expensive. Strategies for reducing transfusion requirements can be divided into preoperative, perioperative and postoperative. The purpose of this study was to investigate our postoperative use of allogenic blood with a view to establish a new transfusion protocol.Patients and Methods  We retrospectively reviewed records of all patients undergoing a primary THR or TKR at Broomfield Hospital between Aug 2004 and Jan 2005 (70 hips, 77 knees). Data recorded included: (i) patient details, (ii) operative details, (iii) preoperative haematological indices, (iv) intraoperative details including length and fluid infusion, (v) postoperative haemotalogical indices, (vi) transfusion requirements and, (vii) length of stay.Results  There was no difference in the mean age between the transfusion and non‐transfusion groups. The mean preoperative haemoglobin (Hb) of THR patients who received a transfusion was significantly lower than those that did not (P = 0.007). This was also found in the TKR patient group (P = 0.035). There was a significant difference in operative time for THR and TKR patients that did not receive a transfusion (U = 565.00, P = 0.02) but no significant difference between transfusion and non‐transfusion groups. The mean postoperative Hb was significantly lower in both the THR (P = <0.01) and TKR (P < 0.01) groups that received a transfusion. There was a significantly higher rate of transfusion in THR patients (49%) as compared to TKR patients (19%). The mean time between the operation and transfusion was 1.7 days for the THR group and 2.5 days for the TKR group. The mean length of stay was significantly longer in both the THR (U = 441.50, P = 0.044) and TKR (U = 194.50, P = 0.001) transfusion groups.Conclusion  The use of allogenic blood can be improved. Two main strategies to achieve this goal include (i) Identifying high risk patients and, (ii) the introduction of a blood transfusion protocol. Identifying high risk patients will not only help preoperative optimisation but also help identify those who should be considered for intraoperative or postoperative cell salvage. The protocol will ensure that those patients who are likely to benefit from a transfusion will receive it as early as possible.
P25 Overnight Transfusions Expose Patients to Unnecessary Risk and Seldom Facilitate Next Day Discharge
Authors: Ambler E. 
Published: September 2006

Aims and Objectives  The Serious Hazards of Transfusion (SHOT) Committee has highlighted the increased risk of overnight transfusion in its annual reports. The risk arises from the paucity of staff (compared to daytime) to monitor patients and manage complications. The darkness on the ward can also obscure early clinical detection of clinical problems. Additionally, patients receiving the transfusion are prevented from sleeping and others in their bay are disturbed throughout the night. The commonest reason offered for overnight transfusion was to enable patient discharge the following day. After seeking approval from the Hospital Transfusion Committee, we performed two ‘snapshot’ studies to investigate the proportion of all transfusions given overnight. Having established that this was a significant number, fifty sequential overnight transfusions were further examined to see whether they were clinically appropriate and whether the recipients were actually discharged the following day. Evidence base/standards. ‘Overnight’ was defined as between 8 pm and 6 am as these are the times between which nursing and medical staffing levels are at their lowest. Transfusions were judged to be appropriate if they were for patients who:• were actively bleeding• admitted symptomatically anaemic during the night• peri‐operativeMethodology  Each morning the Transfusion Practitioner examined the laboratory copies of the transfusion compatibility reports to identify the overnight transfusions. The medical notes for the transfused patients were then consulted for documented evidence as to the appropriateness of each episode according to the criteria above.Results  From the two snapshot studies (each of a month), it transpired that, 43% of units of red cells were given to 40% of the patients and 26% of the units to 42% of the patients. This established that the project would be worthwhile. Of the 50 transfusion episodes, there was documentation in the notes of 30 patients (60%) of the need for transfusion. Seven patients (14%) were noted to be symptomatically anaemic and 3 (6%) bleeding. In total 20% of the overnight transfusions were appropriate by our criteria. Next day discharge occurred in five of the 50 patients. Further work was done to investigate possible delays in commencing the transfusions.Recommendations and actions  The main action taken was to educate staff that inappropriate overnight transfusions compromised their patients’ care and rarely allowed earlier discharge. This was done through re‐enforcement at induction lectures, the Blood Transfusion Policy and a sign on the blood fridge.
P29 Macopharma Prion Capture (P‐CAPT™) Filter Removes Brain‐Derived Prion In Leucoreduced Human RBC
Authors: Gregori L. , Lambert B. C. , Rohwer R. G. 
Published: September 2006

Background  Transmissible spongiform encephalopathy (TSE) diseases (prion) can be transmitted by blood transfusion. Precautionary measures against the spreading of TSE through the blood supply have been implemented, but alone those measures are not sufficient. Pathogen Removal and Diagnostic Technologies (PRDT) developed a resin that reduced brain‐derived TSE infectivity by approximately 4 log10 while endogenous TSE infectivity from hamster blood was reduced to below the limit of detection of the bioassay. The resin was incorporated into membrane layers that were developed by MacoPharma into a prion capture filter termed P‐CAPTTM. In the present study we investigated the performance of the P‐CAPTTM filter to capture PrPres from TSE infected hamster brain homogenate mixed in leucoreduced human RBC.Methods  Hamster brain homogenate infected with the 263K strain of scrapie was added to a unit of leucoreduced (LST1 MacoPharma whole blood filter) human RBC. The unit bag was sterile docked to a P‐CAPTTM kit containing an in‐line P‐CAPTTM filter and a transfer bag to collect the treated RBC. The spiked RBC was applied to the P‐CAPTTM filter by gravity at ambient temperature according to the manufacturer's instructions. At the end of the run, the filter was extensively washed and the captured proteins were analyzed by Western blot. In some studies, two P‐CAPTTM filters and two columns in series were tested to investigate the level of PrPres removal provided by the filter. Other parameters such as reproducibility of binding and filtration time were also analyzed.Results  Western blot analysis of the filter‐bound proteins indicated that the P‐CAPTTM filter captured brain PrPres from a unit of human leucoreduced RBC. The binding was reproducible within the experimental variability of the Western blot signal and the filtration time was also reproducible. The results also indicated that all input PrPres (pathogenic prion protein) was adsorbed by the first filter and no Western blot signal was captured by the second filter in series.Conclusions  P‐CAPTTM filters contain a resin that, when tested in column format, captured brain PrPres, brain‐derived infectivity and endogenous hamster blood infectivity. We showed here that the same resin incorporated into a filter was still capable of capturing brain PrPres.
P33 Transfusion of Incompatible Units to a Patient with Anti‐U: a Case Report
Authors: Lee E. , Redman M. , Taylor C. , Ahmed M. 
Published: September 2006

Introduction  U (MNS5) is a high frequency Ag with the U‐ phenotype blood found mainly in people of Black African origin. Anti‐U has been reported to cause immediate and delayed haemolytic transfusion reactions (HTRs) as well as haemolytic disease of the fetus and newborn (HDFN). The selection of U ‐ RBC is recommended for patients with anti‐U. We present a case of a patient with anti‐U who received serologically incompatible transfusions uneventfully.Case report  A 31‐year‐old black male was admitted to hospital after injury to his right arm. The antibody (Ab) screen was positive and samples were referred to our laboratory for investigation. He grouped as B, D+, U‐ and anti‐U was identified. Due to the severity of the injury the patient underwent emergency surgery without waiting for the Ab ID results. His Hb was 68 g L−1. 5 units of unmatched group O RBC were transfused. During this episode the patient was not given any IVIg cover and there were no clinical or laboratory indications of a transfusion reaction beyond the positive DAT. The patient received four further U‐ RBC transfusions and was treated with parenteral iron.Results

Day
Group
Antibodies
DAT
Hb (g/L)
Tx

Plasma
Eluate
Titre
Subclass
IgG
C3d

1
B
Anti‐U (3+)
NT
8
IgG
Neg
Neg
68
5 O+ U+

2
B
Anti‐U
NT
NA
NT
NT
NT
91
2 B+ U‐

4
B
Anti‐U (w)
Anti‐U (2+)
8
IgG
2+
Neg
99

7
B
Anti‐U (w)
Anti‐U (w)
2
NT
1+
Neg
80
2 O+ U‐

12
B
Anti‐U (2+)
Neg
8
NT
W
Neg
77

19
B
Anti‐U (2+)
Neg
8
NT
Neg
Neg
105

25
NT
NT
NT
NA
NT
NT
NT
117

Discussion  Anti‐U antibodies are generally classed as clinically significant, with the potential to cause immediate and delayed HTRs. When U negative RBC are not available in an emergency, a decision may be taken to transfuse incompatible blood. Close monitoring for any adverse reactions should be undertaken and IVIg cover is often beneficial. In this case, the transfusion of incompatible RBC was uneventful, even without IVIg cover. There was some evidence of an immune process occurring, as over a period of 8 days the Hb fell by 22 g L−1 (22.2%), although the patient did have further bleeding from a wound. An increase in the Hb after day 12 possibly indicates sequestered RBC returning to the circulation, the DAT remained negative. This case report demonstrates that incompatible blood can be successfully transfused in an emergency to a patient with anti‐U. However, severe HTRs due to anti‐U have been reported and so every effort should be made to avoid this situation.