bluuebunny/crossref_metadata_2025 huggingface dataset:
P13 A ‘Blood Transfusion Passport’
Authors: Parris E.
Published: September 2006

It has been repeatedly highlighted in the Serious Hazards Of Transfusion (SHOT) annual reports that the incorrect blood component is often transfused because the blood that was required for the patient was not of the appropriate specification (e.g. should have been irradiated but was not). The Eastern Region is made up of ten NHS Trusts with a wide geographical spread which often involves the need for a patient to share care within the hospitals in this region. Poor communication between health care professionals within hospitals has frequently led to patients not receiving the specialised blood products that they require. The Eastern Transfusion Practitioners Support Group decided to produce a ‘Blood Transfusion Passport’ as a means of improving communication between hospitals on a patients specialist transfusion requirements. The passport is a patient held booklet and was designed along the same principles as the yellow anti‐coagulation booklet. The booklet is currently being trialled within the Eastern region and feedback at the end of the trial will be given to the National Blood Transfusion Committee in the hope that such a Blood Transfusion Passport can be rolled out on a national scale.
P14 Hereditary Persistence of Fetal Haemoglobin Leading to Inappropriate Anti‐D Prophylaxis at Delivery
Authors: Lees S. , Mohamed F. , Devalia V.
Published: September 2006

Fetal maternal haemorrhage (FMH) during pregnancy or delivery can lead to maternal RhD immunisation. Using the Kleihauer‐Betke test, which detects fetal Hb (HbF) in cells, the necessary dose of prophylactic anti‐D can be given to prevent immunisation. Hereditary persistence of fetal haemoglobin (HPFH) is a benign condition where HbF production continues into adult life. Classification is pancellular when HbF levels are elevated in all red cells or heterocellular when HbF is confined to a sub‐population of cells. This is a rare condition, but these HbF cells may be mistaken for fetal cells in the Kleihauer test; we report such a case. Post‐partum samples from an RhD negative woman and her RhD positive infant were tested the day following delivery. 500 iu of prophylactic anti‐D was issued and a Kleihauer test performed to assess FMH volume and additional anti‐D requirement. The FMH was estimated as 33 mL, however the HbFcells appeared as ‘intermediates’ or partially eluted, raising suspicion of HPFH. A total of 4500 iu of prophylactic anti‐D was issued, since although HPFH was suspected, fetal origin of the HbF containing cells could not be excluded due to weekend unavailability of confirmatory testing. Subsequent flow cytometric (FACS) testing for FMH and haemoglobinopathy testing showed the patient to have previously undiagnosed HPFH. The timing of receipt and testing of these samples prevented confirmation of HPFH within the 72‐h deadline for anti‐D maximum effectiveness, leading to administration of precautionary additional anti‐D doses. The likelihood of HPFH was considered because of the appearance of the HbF cells as ‘intermediates’, however not all laboratory staff will be familiar with such cells, particularly if they have not had experience in a haemoglobinopathy laboratory. This possibility should be highlighted.
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‐operative

Methodology  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.
P31 Mixed‐Type Autoimmune Haemolytic Anemia (AIHA)
Authors: Win N. , Tiwari D. , Keevil V. , Needs M. , Lakhani A.
Published: September 2006

The diagnosis of mixed‐type AIHA is based on demonstrating the presence of ‘warm’ IgG auto‐antibody and high thermal amplitude ‘cold’ IgM auto‐antibody reacting at or >30 °C. Mixed‐type AIHA is uncommon. In general, patient with mixed‐type AIHA responds to steroids and reported to be associated with SLE and lymphoma. Awareness of this condition is important as management may be different from either treating warm AIHA or cold haemmagglutinin disease (CHAD). We report two unusual cases of mixed‐type AIHA. Management and outcome of these cases are discussed. An 88‐year‐old, patient was admitted with low Hb and raised bilirubin. He was already on steroids 15 mg daily for joint pain. In view of the strong red cell agglutination in the blood film (CHAD) was suspected initially and were investigated to exclude the secondary causes of CHAD. Three days later Hb dropped to 6.6 g dL−1. Samples were forwarded to the reference laboratory. The DAT was positive with IgG, IgM and C3d. Serological evidence of mixed‐AIHA was confirmed at the reference laboratory. In view of the ongoing haemolysis, steroids were increased to 40 mg daily, and transfusion was given, together with intravenous immunoglobulin (IVIg). Haemolysis ceased and the patient was discharged on decreasing doses of steroid. This case was unusual as the patient developed AIHA while on steroids medication. An 85‐year‐old patient, was admitted with Hb level of 5.7 g dL−1 and evidence of haemolysis. Laboratory investigation confirmed mixed‐type AIHA. A CT scan showed splenomegaly, with small volume lympadenopathy, which were too small to biopsy. A bone marrow biopsy showed no evidence of lymphoma infiltration. The patient was treated with steroids, IVIg, followed by chemotherapy (cyclophosphamide, vincrestine and steroids) with no response. Rituximab (357 mg m−2 weekly for 2 weeks) resulted in no response. The patient received 33 units of red cells over a 9 week period. She underwent splenectomy with resolution of haemolysis and Hb sustained at 11 g dL−1. Splenic biopsy revealed T cell angioimmunoblastic Non‐Hodgkins lymphoma. Splenectomy should be considered in resistant mixed‐type AIHA.
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.
P45 Infotrain: Delivering Information Skills Training Via Web Conference
Authors: Pritchatt D. J.
Published: September 2006

Background  Quality, accessible information is at the heart of continouous quality improvement. Information is only of value when staff can identify and use information to best effect. To do this, staff require information skills, from basic technological skills to higher level discriminatory skills to identify and prioritise evidence‐based resources.National Blood Service (NBS) staff work in widely dispersed locations throughout England, some in blood centres in major cities, others in mobile teams. The new NBS Library Programme is collecting and developing information resources for all staff. A suite of training courses, aimed at a wide range of staff has been developed to raise information awareness, develop information skills and support effective and efficient use of information. To reach and train as many staff as possible, Web Conferencing is being used.

Methods  Travel costs and time spent away from work activities across the organisation currently need to be reduced, so an innovative delivery method was sought. Following initial, wide‐ranging consultation a core of topics were identified, to appeal to the widest possible number of staff. NBS had recently purchased Web Conference software and had existing access to teleconferencing. This enabled Web Conference delivery to be trialled. A series of courses (InfoTrain) was developed, consisting of short sessions to suit this mode of delivery. Courses were offered widely across the organisation.

Results  Evidence of effectiveness is being collated from questionnaires, on a database. Verbal/email feedback, course bookings and invitations to deliver sessions at other blood centres are further indicators. Initial results show courses directly contribute to staff's work. Demand for courses, e.g.‘Hierarchy of Evidence’and ’Electronic Journals’ has led to waiting lists being established. Trainees have indicated increased awareness of available resources and information quality issues.

Conclusions  Courses designed to suit Web Conferencing can be very successful and widely welcomed. Web Conferencing significantly reduces travel costs and time spent away from core work activities. Information skills training is of demonstrable benefit to staff across the organisation .Training increases independence in information seeking; raising awareness of information quality issues and guiding staff to the best resources can benefit the whole organisation, by reducing time spent identifying and accessing the required information. Increasing staff's understanding of evidence based healthcare impacts directly upon practice, enabling independent assessment of personal practice against evidence and guidance, leading to continuous service improvement.
P48 Molecular Characterisation of the Variable Domains of an ΑIIbβ3 Specific IgM κ Platelet Cold Agglutinin in a Follicular Lymphoma Patient with Treatment Refractory Thrombocytopenia
Authors: Jennings N. S. , Harmer I. J. , Campbell K. , Smith G. A. , Stafford P. , Metcalfe P. , Benton M. A. , Marsh J. C. W. , Ouwehand W. H.
Published: September 2006

Cold agglutinins (CA) were originally defined as IgM κ antibodies capable of agglutinating erythrocytes at temperatures below 37°C and cold haemagglutinin disease may ensue in patients with high titre cold agglutinins at low environmental temperature. Platelet CA are rare and have been poorly characterised. We present the first molecular characterisation of a pathological, platelet reactive, CA. This antibody was identified in a patient with follicular lymphoma (FL) who had profound, refractory, thrombocytopenia that resolved in response to Rituximab. Serological and molecular studies of this IgM κ that recognises αIIbβ3 shows that binding is temperature dependent but independent of the integrin activation state. The VH1‐02 and Vλ3‐20 encoded variable domains of the heavy and light chains have characteristics typical of a low affinity autoantibody. Maldi‐Tof analysis of the platelet reactive IgM showed unequivocally that the IgM was derived from the FL clone. Interestingly we observed idiotypic cross‐reactivity between this IgM paraprotein, several other FL idiotypes and our β3‐leucine 33 (HPA‐1a) specific human recombinant antibody. Our results suggest that binding of the latter and other αIIbβ3 specific monoclonal antibodies induces a subtle conformational change that enhances platelet binding of the patient IgM.
P49 First UK Report of a Case of Glycoprotein VI Deficiency in a 33‐Year‐Old Patient with Moderate Thrombocytopenia and Autoantibodies Against the Immunoglobulin‐Like Domains of the Receptor
Authors: Smethurst P. A. , Gladigau E. L. , Garner S. F. , Davies L. W. , Rankin A. , Campbell K. , Yusuf R. , Farndale R. W. , Tillyer L. , Ouwehand W. H.
Published: September 2006

Background  Studies of cases of acquired and inherited GPVI deficiency have made critical contributions to the understanding of the pivotal role of GPVI in collagen mediated platelet signaling.

Aim  To characterize the first UK case of GPVI deficiency.

Methods  Numerous platelet function tests and immunophenotyping assays, western blotting and genomic sequencing.

Results  A female patient of 33 years with mild thrombocytopenia (80–120 × 109 L−1), had experienced moderate bleeding with atypical locations, including bruises and petechiae on neck and chest, and a perineal haematoma and postnatal bleeding. Bone marrow examination revealed plentiful megakaryocytes, compatible with a state of compensated thrombocytolysis. PFA 100 response with the collagen/epinephrine cartridge was >295 seconds, and aggregometry showed a profoundly reduced response to collagen and collagen‐related peptide. Response to ADP and PAR1 peptide was moderately reduced, but ristocetin induced agglutination was normal. By flow cytometry, normal surface expression was observed to many platelet receptors, including GPIIbIIIa, GPIaIIa and PECAM‐1. Expression of PAR‐1 was mildly reduced, whereas for GPVI a 90% reduction in surface expression was observed. Total GPVI levels as measured by blot densitometry were also reduced, although not as profoundly. The sequence of the GP6 exons was identical to the reference sequence. A potent GPVI autoantibody was detected in the patients’ plasma by a sandwich ELISA (MAIPA), and reactivity was mapped to the collagen‐binding domains using recombinant GPVI fragments.

Conclusions  The patient has GPVI autoantibodies of the IgG class, resulting in an acquired GPVI deficiency and with a bleeding phenotype apparently more severe than other reported cases of GPVI deficiency.
P05 Distribution and Significance of High Titre Anti A and Anti B in a UK Blood Donor Population
Authors: Lattimore C.
Published: September 2006

A risk of clinically significant haemolysis exists where high titre (HT) anti A and anti B in blood components, particularly platelets, are transfused to patents with the corresponding antigen. Risk is reduced by screening all donations and designating packs with an anti A or B titre of over 1:128 as HT positive. This study reports on agglutination titres, haemolysin levels and the sex of donors. The Olympus PK7200 is used for automated screening of donors; 20 HT positive, 20 HT query and 20 negative group O donations were selected. Agglutination titres against A1 and B cells were determined manually by incubating one drop of doubling dilution of donor plasma from neat to 1 : 512 at room temperature for two hours with one drop of cells. All samples were manually tested for the presence of haemolyisns; one drop of washed A1 or B cells and four drops of neat donor serum were incubated at 37 0C for 5 min, centrifuged and visually inspected for haemolysis. In addition, the sex of the donor was noted from session records. All HT samples had a manual titre between 1 : 128 and 1 : 512, with levels of agglutination between one and five. The HT query samples showed agglutination below 1:128 and the HT negative samples showed agglutination below 1 : 32. Haemolysin testing showed hemolytic activity in 16/20 HT positive samples, 4/20 HT query and 1/20 HT negative. Of the HT positive donors 18/20 were female, as were 13/20 HT query and 11/20 HT negative. The results of the manual titres suggest the Olympus PK7200 is accurate in detecting HT positive and query donations. The results of the haemolysin investigation show good correlation with HT testing results. Findings of gender analysis support a theory of female donors having more HT antibodies, possibly as a result of previous stimulating events. Guidelines for ‘out of group transfusion’ and associated donor testing varies internationally. An important consideration is balancing the risk of transfusion reaction against the risk of limited availability of group matched products. Reports of haemolytic reactions are uncommon but SHOT reports repeatedly demonstrate ongoing cause for concern.
P58 The Novel Tyrosine Kinase Inhibitor Nilotinib is Equipotent with Imatinib and Does Not Induce Apoptosis in Chronic Myeloid Leukaemia Stem Cells
Authors: Allan E. K. , Jorgensen H. G. , Holyoake T. L.
Published: September 2006

The novel tyrosine kinase inhibitor nilotinib is equipotent with imatinib and does not induce apoptosis in CML stem cells Chronic myeloid leukaemia (CML) is a myeloproliferative disorder of stem cell origin characterised by a shortened ‘Philadelphia’ (Ph+) chromosome 22. A novel oncoprotein BcrAbl is translated from the fusion oncogene, bcrabl, that arises from the t 9;22 reciprocal translocation. BcrAbl is a constitutively active tyrosine kinase that upregulates diverse intracellular signal transduction pathways resulting in multiple adverse effects on cell processes including proliferation, differentiation and adhesion. The rationally designed tyrosine kinase inhibitor, imatinib mesylate (IM; Glivec®, Novartis Pharma, Switzerland) has proven highly effective in the treatment of CML with the majority of early chronic phase patients achieving complete cytogenetic remisssion and in some cases molecular remission. However, many advanced phase patients have developed clinical resistance. Previous in vitro studies by our group have highlighted a population of quiescent Ph+ stem cells that persist and accumulate following IM exposure demonstrating the anti‐proliferative effects of this drug. Nilotinib, (AMN 107; Novartis Pharma) is a novel Abl tyrosine kinase inhibitor specifically designed to be more selective for BcrAbl with activity against IM‐ resistant mutations. In the Ph+ K562 cell line, nilotinib's IC50 was 30 ± 10 nm versus 600 ± 60 nM for IM consistent with its reported 20‐fold increased potency. However, in primary CD34+ CML cells, nilotinib and IM were equipotent for inhibition of BcrAbl activity, with 5 μM of either compound producing equal but incomplete reduction in Crkl phosphorylation. CD 34+ cells were able to expand over 72 h with either drug up to 5 μM although there was a concentration dependent restriction of amplification. These results confirmed an anti‐proliferative rather than pro‐apoptotic effect of nilotinib. Morever, the most primitive cells persisted and accumulated over 72 h with nilotinib and remained caspase‐3 negative. Combination of IM with nilotinib led to increased accumulation of this population suggesting at least additive anti‐proliferative effects. Thus despite activity against IM resistant mutants, nilotinib may not improve clinical responses at the stem cell level with respect to IM.
P06 Use of Intraoperative Red Cell Salvage During Elective Abdominal Aortic Aneurysm Repair is Safe and Associated with Reduced Need for Allogeneic Blood Transfusion.
Authors: Singh‐Ranger G. , Fayaz A. , Leppard T. , Khan Y. , May A. R. L. , Backhouse C. M.
Published: September 2006

Introduction  The need for blood transfusion during vascular surgery can be considerable. Concerns regarding the safety and cost of allogeneic blood have demanded scrutiny of alternative methods of blood conservation during these operations, including intraoperative red cell salvage. Autotranfusion is purported to reduce the need for blood transfusion, and therefore its attendant complications.

Method  We retrospectively collected data on all patients undergoing elective infrarenal abdominal aortic aneurysm repair in our unit, and compared patients in which the cell saver was used to those in which it was not, in terms of operative blood loss, transfusion requirement, length of stay, and postoperative haemoglobin. The Mann Whitney test and Kendall's tau B statistics were used to analyse the data

Results  Thirty‐five patients underwent AAA repair with use of the cell saver, and 31 patients underwent repair without the use of this device. We found a significant association between the use of the cell saver and reduced need for allogneic blood in these patients (Table). There was also a significant positive correlation between the volume of blood obtained by the cell saver, and that returned to the patient (t = +0.5, P < 0.00001). The postoperative haemoglobin in the cell saver group was significantly lower, but still within acceptable range. There was no significant difference in mortality between the two groups;

Variable
Cell Saver Group ‐Mean
No‐Cell Saver Group ‐ Mean
Significance

Age (years)
72.4 (8.1)
74.5 (7.1)
0.11

AAA size (cm)
6.2 (0.9)
6.1 (0.7)
0.94

Preop Hb (g/dL)
13.9 (1.2)
13.9 (1.6)
0.92

Operative blood loss (mL)
986.7 (494.2)
1900.6 (1657.6)
0.02

Intraoperative allogeneic blood used (no. units)
0.5 (1.2)
2.6 (2.8)
<0.0001

Total allogeneic blood (7d)
0.9 (1.4)
3.5 (4.3)
<0.0001

Post operative Hb
10.9 (1.2)
11.7 (1.7)
0.03

Length of stay (days)
12.5 (4.2)
10.7 (8.1)
0.01

Conclusion  Our study indicates that use of the cell saver in elective AAA repair is associated with a reduced need for allogeneic blood transfusion, and therefore, with a reduction in risks of transmitted infection and transfusion reaction. These findings also have important financial implications.
P60 Irradiated Skin Allografts: a New Product for Clinical Use?
Authors: Hogg P. , Lomas R. , Rooney P. , Eagle M. , Kearney J.
Published: September 2006

Introduction  There is an increasingly elevated demand for cryopreserved skin allografts in the UK. However current antibiotic disinfection of viable skin allografts results in an approximately 20% microbial failure rate. High concentrations of glycerol have been shown recently to maintain the functional properties of terminally irradiated skin. The aim of this study was to investigate whether γ irradiation in combination with glycerol treatment could salvage the failed skin and allow it to be released for clinical use.

Methods  Skin samples from three donors with consent for research that had been treated with 20% glycerol and processed into viable cryopreserved allografts were treated as below (Table 1).

Group
Solute
Treatment
Temperature

1
20% glycerol (control)
None
‐80°C

2
20% glycerol
25kGy γ irradiation
‐80oC

3
Phosphate buffered saline
25kGy γ irradiation
‐80oC

4
50% glycerol
25kGy γ irradiation
‐80oC

5
85% glycerol
25kGy γ irradiation
‐80oC

6
85% glycerol
25kGy γ irradiation
Ambient

The skin samples were then evaluated for their histological structure, in vitro cytotoxicity and suitability for clinical use. The anti‐microbial activity of the different concentrations of glycerol alone against Bacillus pumilis spores with and without radiation was also assessed.

Results Irradiation affected the cytotoxicity and physical properties of skin in different ways (Table 2). The skin from group 2, which was irradiated in 20% glycerol at ‐80 oC was found to be the best preserved. All the concentrations of glycerol, which had been irradiated, killed the Bacillus pumilis apart from 85% glycerol at ‐80 oC.

Group
Histology
Cytotoxicity
Clinical Use

1
Normal
None
Good‐ would use

2
Normal
None
Good ‐ would use

3
Damage
Some
Would use

4
Normal
Some
Less pliable ‐would use

5
Damage
Some
Less pliable ‐would use

6
Damage
Extensive
Less pliable ‐would use

Conclusion  We can infer from the results that high concentrations of glycerol confer radioprotection on skin but also microorganisms as well. Whereas 20% glycerol leaves the biological characteristics of skin unchanged and kills the microorganisms. The implications of this study are that cryopreserved irradiated grafts can be made available for surgical use when cryopreserved viable grafts are not available or required.
P63 Stem Cell and Immunotherapy Services, NHS Blood and Transplant, Birmingham, UK
Authors: McDonald D. , Deavall H. , Moss P. , Steele J. , Thompson J. , Turner V.
Published: September 2006

Over 20 years ago, the Birmingham Blood Centre established a facility for the cryopreservation of bone marrow (BM) for patients in the West Midlands suffering from haematopoietic disorders and for whom a bone marrow transplant was indicated. Today, the use of mobilised peripheral blood (PBSC) has overtaken bone marrow as the source of stem cells for transplantation and the numbers of patients benefitting and the diversity of conditions being treated has increased enormously. Allogeneic transplants, using stem cells from healthy donors, have become increasingly successful as a result of an improving understanding of the complexities of the HLA histocompatibility system. Additionally umbilical cord blood (HUC), which in the 1980s was recognised as a source of stem cells, can now be collected and used for transplantation. As scientific knowledge and the clinical management of patients has advanced, so too have laboratory methods for manipulating cell products to enrich or deplete certain cell populations (e.g. by CD34+cell selection) in order to minimise potentially fatal graft‐versus‐host disease (GVHD) or to eliminate tumour cells in the case of autologous patients. Donor lymphocytes (DLI) may also be collected and used to aid a graft‐versus‐leukaemia (GVL) effect. The laboratory is currently developing protocols for immunotherapy using virus‐specific T cells which can be prepared and infused to combat potentially fatal CMV disease post‐transplant. Clinical trials of vaccines employing tumour specific dendritic cells for treating patients with hepatocellular carcinoma (HCC) and metastatic melanoma (MM), which do not respond to conventional treatments, are also underway. The advances and expansion in the Stem Cell and Immunotherapy (SCI) service in Birmingham over the last 10 year period are reflected in the table below:

Year
Stem Cell Products received by laboratory
Manipulated Stem Cell Products

Total
BM
PBSC
HUC
Auto
Allo
CD34+ selection
DLI

1995
409
76
333
0
400
9
2
1

2005
670
16
654
10
546
124
20
40

The laboratory is fully accredited by the Human Tissue Authority (HTA), MHRA and JACIE. It occupies new purpose built facilities with the most up to date equipment. Patients in the West Midlands awaiting an haematopoietic stem cell transplant or cellular therapy can be assured of receiving treatment equivalent to the best that is available anywhere in the world.
P07 Reduction of Allogeneic Red Cell Transfusion using Autologous Blood Cell Salvage in Knee Arthroplasty
Authors: Hall R. , Johnson J. , Goudie K. , Clark M. , Chambers J. , Senior C. , Hartley R.
Published: September 2006

Recent Hospital Transfusion Committee (HTC) audit at the Royal Bournemouth Hospital (RBH) confirmed an allogeneic red cell transfusion rate of 20% for primary Total Knee Replacement (TKR). Current policy at RBH states that when blood stocks reach 67% of normal (amber alert) then surgery with a >20% likelihood of blood transfusion will be cancelled. At current transfusion rates this would include primary TKR. Recent studies have shown a reduction in allogeneic transfusion rates when autologous transfusion drains are utilized. The purpose of this study was to see whether the current rate of allogeneic transfusion could be reduced with the introduction of the CellTransTM Autologous Knee Drainage Blood Transfusion System (ABT) in TKR at RBH. Over a 3 month period all patients undergoing primary, bilateral or revision knee arthroplasty received an ABT. Demographic data was collected from the orthopaedic pre‐assessment clinic. Following surgery further data was collected relating to volume of blood loss into the drain, volume of autologous blood re‐transfused, units of allogeneic blood required and the transfusion trigger, postoperative haemoglobin levels, infection rates and length of stay in hospital. We then compared this data set with retrospective data. Of 170 patients undergoing knee arthroplasty 141 received the ABT. The data collected was compared retrospectively with 169 patients from the previous 3 month period. We demonstrated a reduction in transfusion rates of 13% for primary TKR, 42% for bilateral TKR and 57% for revision TKR with the use of the ABT. In addition we demonstrated a reduction in total allogeneic blood use (99 units to 26 units) and a reduction in mean length of stay in hospital (8.6 days to 7.5 days) with the ABT. Further analysis of the data collected showed a 46% reduction in the allogeneic transfusion rate and a reduction in total allogeneic blood usage (99 units to 9 units) of anaemic patients presenting for surgery. This study has demonstrated a dramatic reduction in allogeneic blood transfusion rates with the use of the CellTransTM Autologous Blood Transfusion System. We have also shown a reduction in length of stay in hospital. Prior to the study primary total knee replacement would have been cancelled during times of limited blood availability (amber alert). The use of the ABT is good for the patient in reducing the need for allogeneic blood, and in addition has demonstrated a significant cost saving due to the reduced blood usage and potential prevention of cancelled operation lists.
Cohort‐dependent seedling recruitment, survival and reproductive capacity of Tribulus terrestris
Authors: VERDÚ A M C , MAS M T
Published: September 2006

Knowledge of the population biology of the summer annual Tribulus terrestris can provide valuable information for managing the risk of its colonising crop fields. Because of spring‐summer crop management practices, fields can exhibit bare ground in late April and early May. In this situation, four cohorts in 2000 and three cohorts in 2001 were censused and monitored throughout the vegetative and reproductive season, in order to evaluate the relationship between time of emergence, survival, vegetative growth and plant reproductive capacities. To a great extent, seedling survivorship depended on emergence timing; the earlier the emergence of the seedlings, the higher was the proportion of survivorship (64% and 4%, for early and late emergences). The time of emergence also influenced plant development; in both years, surviving individuals of the first cohorts presented significantly greater plant biomass and larger numbers of reproductive structures (60.5 fruits per plant) than those of later cohorts (0.8 fruits per plant). Reproductive meristem allocation was also highest in the earliest cohorts. Results showed that T. terrestris can increase its population size in fields if gaps of bare soil surface are available from mid spring to early summer.
L/I‐5 Hemodynamics in adult‐to‐adult living donor liver transplantation: personal experience
Authors: Nadalin S. , Li J. , Radtke A. , Sotiropoulos G.C. , Malamutmann E. , Broelsch C.E. , Malagó M.
Published: September 2006

Introduction:  The interaction and modulation of inflow and outflow in adult living donor liver transplants (LDLTs) is very complex; small‐for‐size syndrome (SFSS) still represents the most dangerous complication in this regard.

Methods:  We measured the liver hemodynamics, in both donors and recipients, in 21 LDLTs.

Measured parameters included hepatic artery flow (HAF) and portal vein flow (PVF) (ml/min/100 g ), PVF/HAF ratio (PAR) by means of transit time flowmeter, and portal vein pressure (PVP). For all recipients, we used a maximalized outflow with blanket technique (with MHV in 19). According to the results, in order to prevent SFSS, we performed either splenic artery ligation (SAL) or modified porto‐caval‐shunt (PCS) or intrarterial PGI2 + steroid infusion.

Results:  The median reference values in donors were as follows: HAF, 26.5 ml/min/100g; PVF, 78.5 ml/min/100g; and PAR, 2.5. At total of 5 patients underwent SAL because of GBWR ≤ 0.9; HAF decreased from 32% to 82% in 4; PVF increased from 300% to 400% in 3; PAR was > 20 in 2. None of these patients developed SFSS. Two patients underwent intraarterial infusion of PGI2 + steroids over 14 days because of a PVF increase ≥ 300%, HAF decrease of 82% in 1, PAR 66 and 22, PVP > 20 in 1, and impossibility of SAL. No SFSS was observed. One patient underwent a modified PCS because of PVF increase of 250%, HAF decrease of 70%, PAR 27. No complications occurred.

Discusssion:  Strength monitoring of liver hemodynamics at pre‐, intra‐, and posthepatic level allows specific management at the different levels in order to prevent SFSS and graft loss.
Genomic organization, localization and polymorphism of porcine PSMB10, a gene encoding the third beta‐type proteasome subunit of 26S proteasome complex
Authors: Wu X. , Yang S.L. , Yerle M. , Zhu Z.M. , Wang H.L. , Wang H. , Li K.
Published: September 2006

The proteasome subunit, beta type 10 (PSMB10) gene regulated by interferon‐gamma is a core part of the 26S proteasome complex, which is an important protein degrading system. Isolation and characterization of swine PSMB10 revealed a conserved structure with other mammalian PSMB10 genes. An A/G nucleotide polymorphism in PSMB10 intron 2 and a C/T single nucleotide polymorphism in exon 5 were detected by polymerase chain reaction restriction fragment length polymorphism. The allele frequencies were significantly different among Tongcheng, Landrace, Large White and Duroc. The porcine PSMB10 was mapped by somatic cell hybrid panel and radiation hybrid mapping on SSC6p14–p15, which is in good agreement with human–pig comparative maps.
Identification of single‐nucleotide polymorphisms in 5′ end and exons of the PRKAG3 gene in Hubbard White broiler, Leghorn layer, and three Chinese indigenous chicken breeds
Authors: Zhao C.J. , Wang C.F. , Deng X.M. , Gao Y. , Wu Ch.
Published: September 2006

5′‐AMP‐activated protein kinase plays an important role in regulating the level of ATP in the presence of metabolic stress. Previous studies revealed that polymorphisms in 5′‐AMP‐activated protein kinase gamma 3 subunit (PRKAG3) gene are associated with meat quality in pigs. In the present study, single‐nucleotide polymorphisms (SNPs) in the 5′‐end and exons of chicken PRKAG3 gene were identified with the method of single‐strand conformation polymorphism in Hubbard ISA White broiler, Leghorn layer, and three Chinese indigenous chicken breeds, Tibet Chicken, Shouguang Chicken and Beijing Yellow Chicken. Two SNPs in the 5′‐end of the gene and 10 SNPs in exons 3, 4, 9 and 11, of which three caused amino acid substitutions, were identified in the PRKAG3 gene of the five chicken breeds. The results will facilitate further study on the association between the mutations of PRKAG3 and chicken meat quality.