bluuebunny/crossref_metadata_2025 huggingface dataset:
Effectiveness of written guidelines on the appropriateness of thromboprophylaxis prescriptions for medical patients: a prospective randomized study
Authors: FONTAINE A. , MAHÉ I. , BERGMANN J. F. , FIESSINGER J. N. , DHOTE R. , COHEN P. , VINCENEUX P.
Published: September 2006

Objective.  To assess the effectiveness of providing doctors with written thromboprophylaxis prescription aids based on current recommendations.

Design.  A prospective trial of specific anticoagulant prescription forms: a 1‐day survey before and after the intervention in each centre.

Setting.  30 Internal Medicine departments of Assistance Publique‐Hôpitaux de Paris.

Subjects.  All inpatients were included, except those who were either admitted or discharged on the day of the survey, and those receiving curative anticoagulant treatment.

Interventions.  The study included three parts: (i) a 1‐day baseline survey; (ii) over the following 3‐month period, departments were randomized into two groups: all practitioners in wards allocated to the intervention group were required to systematically use specific anticoagulant prescription forms, whilst doctors in the control group continued prescribing according to their usual practices and (iii) a 1‐day postintervention survey.

Main outcome measure.  The proportion of prescriptions in accordance with the recommendations.

Results.  1469 patients were included. The intervention produced a significant reduction in the frequency of over‐prescriptions, from 25% to 14% of the patients who did not meet the guideline criteria (adjusted OR: 0.3; 95% CI: 0.1–0.8). Using specific forms did not improve under‐prescription of anticoagulants. A little over 60% of the patients who met guideline criteria for thromboprophylaxis were prescribed anticoagulants in both intervention and control wards, either at baseline or after intervention.

Conclusions.  Multitargeted interventions using a variety of means and strategies should still be considered to improve prescriptions that may have a significant impact on health expenses and, most importantly, on patients outcomes.
Evaluation of a primary care‐oriented brief counselling intervention for obesity with and without orlistat
Authors: POSTON W. S. C. , HADDOCK C. K. , PINKSTON M. M. , PACE P. , REEVES R. S. , KARAKOC N. , JONES P. , FOREYT J. P.
Published: September 2006

Objective.  There is a significant need for an obesity treatment model suitable for the primary care environment. We examined the effectiveness of a brief counselling intervention alone, in combination with orlistat, and drug‐alone in a 12‐month randomized‐clinical trial at a medical school obesity centre.

Methods.  Participants (N = 250) with body mass index (BMI) ≥27 were randomized. Changes in body weight, lipids, blood pressure and serum glucose were examined. Drug adherence and attendance were also evaluated.

Results.  Completers analysis was conducted on 136 participants with data at baseline, 6 and 12 months and intention‐to‐treat analyses (ITT) for the total sample. Amongst completers, participants in the drug only (P = 0.012) and drug + brief counselling (P = 0.001) groups lost more weight (mean ± SD: −3.8 ± 5.8 kg and −4.8 ± 4.4 kg, respectively) than participants in the brief counselling only group at 6 months (−1.7 ± 3.3 kg), but there were no significant group differences at 12 months. ITT model results were similar to completers at 6 months and remained significant at 12 months, but the weight losses were more modest (<3 kg) for both groups receiving orlistat. For brief counselling alone, participants gained weight (1.7 ± 4.2 kg). Cardiovascular disease (CVD) parameter changes were negligible.

Conclusions.  Pharmacotherapy alone or combined with brief counselling resulted in modest weight losses that had minimal impact on cardiovascular parameters, but were greater than brief counselling alone. Whilst brief interventions and primary pharmacotherapy have been suggested as viable treatments for implementation in primary care settings, our study suggests that such minimal interventions provide minimal benefits.
Metabolic Signals Modulate Hypothalamic‐Pituitary‐Adrenal Axis Activation During Maternal Separation of the Neonatal Mouse
Authors: Schmidt M. V. , Levine S. , Alam S. , Harbich D. , Sterlemann V. , Ganea K. , De Kloet E. R. , Holsboer F. , Müller M. B.
Published: September 2006

The postnatal development of the mouse is characterised by a period of hypo‐responsiveness of the hypothalamic‐pituitary‐adrenal (HPA) axis to moderate stressors. Maternal separation disinhibits this blockade of the HPA axis, but the mechanism responsible is not clear. The present study examined the influence of metabolic signals on the central and peripheral components of the HPA axis in neonatal mice aged 8 days in absence or presence of the mother. Reductions in plasma glucose and leptin as well as rapid increases in plasma ghrelin were apparent in the neonate 4 h following maternal deprivation and maximal at 8 h. In addition, maternal separation induced an increase of neuropeptide Y (NPY) mRNA expression in the arcuate nucleus, a decrease of corticotrophin‐releasing hormone (CRH) mRNA expression in the paraventricular nucleus and a rise in serum corticosterone. Pharmacological manipulation of the metabolic signals attenuated the HPA response to maternal separation. Thus, the rise in plasma corticosterone induced by maternal separation was ameliorated by prevention of reduction in blood glucose or blockade of the ghrelin signalling pathway, as were the hypothalamic changes in NPY and CRH mRNAs. By contrast, leptin treatment did not affect the HPA axis response to maternal separation. Together these results suggest that metabolic signals play an important role in triggering the HPA response of the neonate to maternal separation.
Identification and characterization of receptors for vacuolating activity of subtilase cytotoxin
Authors: Yahiro Kinnosuke , Morinaga Naoko , Satoh Mamoru , Matsuura Gen , Tomonaga Takeshi , Nomura Fumio , Moss Joel , Noda Masatoshi
Published: September 2006

Some shiga toxin‐producing Escherichia coli secrete a novel AB5 cytotoxin, named subtilase cytotoxin (SubAB), which induces vacuole formation in addition to cytotoxicity in susceptible cells. By immunoprecipitation with SubAB from Vero cells, we discovered proteins of 100 kDa, 135 kDa and 155 kDa as potential candidates for its receptor. These proteins were N‐glycosylated in their extracellular domains, a modification that was necessary for interaction with SubAB. Biotinylated receptors were partially purified by Datura stramonium agglutinin affinity chromatography and avidin‐agarose and analysed by TOF mass spectroscopy. The peptide sequences of p135 were identical to β1 integrin, and its identification was confirmed with anti‐integrin β1 antibody. The p155 protein was identified as α2 integrin using anti‐integrin α2 antibody. In addition, treatment of Vero cells with β1 integrin RNAi before exposure to SubAB prevented vacuolating activity. These results suggested that SubAB recognizes α2β1 integrin as a functional receptor; this first interaction may be an important key step leading to the SubAB‐induced morphological changes in Vero cells.
Defining the Mga regulon: comparative transcriptome analysis reveals both direct and indirect regulation by Mga in the group A streptococcus
Authors: Ribardo Deborah A. , McIver Kevin S.
Published: September 2006

The regulator Mga in the group A streptococcus (GAS) is known to directly activate several virulence genes important for colonization and immune evasion. Transcriptome analysis comparing two mga‐1 serotypes (M1 SF370, M6 JRS4) and one mga‐2 serotype (M4 GA40634) against their isogenic mga‐inactivated strains uncovered a broader Mga regulon profile containing both activated and repressed genes with predicted functions primarily related to sugar metabolism. This was reflected in the altered abilities of M1 and M4 Mga mutants to grow in chemically defined media with a single sugar source compared with their wild‐type counterparts. Although the M1 and M4 Mga profiles were similar, the M6 JRS4 was clearly distinct, even from other M6 strains. Real‐time RT‐PCR and Northern blots confirmed that established core Mga regulon genes directly activated by Mga (emm, scpA, sof, fba) exhibited the highest activation levels across all strains tested. Spy2036 encoding a cytosolic hypothetical protein was highly activated in all three serotypes and was called gene regulated by Mga (grm). Mga bound directly to Pgrm, which overlaps the Mga‐regulated Psof in OF+ strains, suggesting that grm is part of the core Mga regulon and Mga is able to activate divergently transcribed genes from a single site. Furthermore, Mga activated speB when detectable in the wild‐type strain, although direct binding of Mga to PspeB could not be demonstrated. Thus, Mga is able to both directly and indirectly regulate genes shown to be important for virulence and the metabolic homeostasis of GAS.
Immune modulation by helminthic infections: worms and viral infections
Authors: KAMAL S. M. , EL SAYED KHALIFA K.
Published: September 2006

Helminthic infections occur worldwide, especially in developing countries. About one‐quarter of the world's population, 1·5 billion, are infected with one or more of the major soil‐transmitted helminths, including hookworms, ascarids, and whipworms. Schistosomes infect more than 200 million people worldwide with 600 million at risk in 74 countries. The interaction between helminths and the host's immune system provokes particular immunomodulatory and immunoregulatory mechanisms that ensure their survival in the host for years. However, these changes might impair the immunological response to bystander bacterial, viral, and protozoal pathogens and to vaccination. Modulation of the immune system by infection with helminthic parasites is proposed to reduce the levels of allergic responses and to protect against inflammatory bowel disease. In this review, we summarize the immunological milieu associated with helminthic infections and its impact on viral infections, mainly hepatitis B virus, hepatitis C virus, and human immunodeficiency virus in humans and experimental animals.
PL01 Background to Testing Deceased Tissue Donors for Variant CJD
Authors: Warwick R.
Published: September 2006

Introduction  Hundreds of thousands of patients in Europe undergo therapeutic human allograft treatments. There have been three recent reports of possible transmission of vCJD or of abnormal prion by blood transfusion. There is no validated blood test for vCJD, but there are techniques using tissues, that have been used in vCJD epidemiological studies. The principle of testing tonsil, spleen or other reticulo‐endothelial or neurological tissues deserves discussion including issues of test validation, logistics of biopsy sampling, timeframes, potential implications for other human material procurement and ethical issues.

Method  The advantages and disadvantages of instituting vCJD tests on deceased donors was considered, legal advice sought and logistical issues considered.

The advantages of undertaking vCJD testing of tonsil  (i) Possibility of identifying potential vCJD deceased donors in the late latent phase of vCJD infection; and (ii) UK legal precedence suggests that the public is legitimately entitled to expect that blood, and presumably tissues, are 100% safe.

Disadvantages to testing  (i) Unknown time when in the pre‐clinical phase abnormal PrP may be detectable; (ii) The sensitivity of tests may be less than required; (iii) The impact of false positive and false negative results (iv) Logistical aspects of testing to GMP standards; (v) Impact of laboratory turnaround time for any PrP test on tissues/organs requiring rapid release; (vi) Possible pre‐disposition to vCJD disease impact on counselling donor families; (vii) Implications for lookback on recipients an infected tissue donor where previous donations may have been given, either of blood or tissues; (viii) Risk of contamination of tissues or processing laboratories from biopsies; (ix) Loss of categories of tissue donors due to unwillingness of donor families to consent to testing; and (x) Loss of tissue donors from living donor programmes where tonsil testing could not be undertaken.

Results  It was concluded that tonsil and splenic biopsies should be investigated logistically to provide analytes for vCJD testing.

Conclusion  Multidisciplinary work is underway to take forward vCJD testing of deceased donors.
SI07 High‐Throughput Genotyping for Red Cell and Platelet Blood Group Antigens by DNA Micro Arrays
Authors: De Haas M. , Van Der Schoot C. E. , Maaskant‐van Wijk P. A.
Published: September 2006

In the Netherlands, to provide antigen‐negative red cells for certain groups of patients, the majority of donors is serologically typed for ABO, RhCcDEe and K and a subset of donors for clinically relevant systems (e.g. Fy, Jk, MNSs). Complete phenotyping of all blood donors, for all blood group antigens including high frequency antigens, is too laborious and simply not feasible, because of the lack of sufficient and high‐quality typing reagents. For most blood group systems the molecular basis is known and found to be a single nucleotide polymorphism (SNP). In the last years, methods were developed facilitating high‐throughput blood group genotyping by glass‐based DNA microarrays. By this so‐called chiptechnology the phenotype of a donor for all clinically relevant (including HFA) red cell antigens can be simply predicted by running a single assay. Typing of platelet antigens and perhaps also HLA can be included as well. Therefore, high‐throughput genotyping or chiptechnology promises the future availability of a completely typed red cell and platelet inventory. A disadvantage of genotyping is false positive or negative results, due to currently unidentified or newly occurring mutations leading to null alleles or mispriming. Large‐scale studies of blood group antigen typing including different ethnic groups are required to determine the error rate. An advantage of the use of blood group genotyping assays is improved detection of ‘weak phenotypic signals’, for example weak‐D or Fyx antigens will be correctly identified by genotyping. If all red cells units are routinely completely typed a less restricted policy on preventive matching for clinically relevant blood group systems can be implement. Thus, in more patients or even in all blood recipients it may become possible to avoid alloimmunization with the benefit of a reduction in the occurrence of severe transfusion reactions (for example delayed haemolytic transfusion reactions by Jka alloantibodies) or a decreased survival time of transfused cells. Furthermore it will reduce costs and delays in provision of antigen‐negative red cells. Thus, if the challenge can be met to develop the current microarrays or chiptechnology into systems that meet the throughput and quality demands of the blood bank, this can lead to an important change in transfusion policy.
SI09 Analysing Platelet Function in Blood Donors: Selection of Subjects for Functional Genetic Studies
Authors: Garner S. F. , Jones C. I. , Angenent W. , Bernard A. , Carr P. , Hogwood J. , Rankin A. , Stephens J. , Tom B. D. , Walton J. , Dudbridge F. , Ouwehand W. H. , Goodall A. H.
Published: September 2006

Introduction  The objective of this study; performed as part of the BLOODOMICS project, was to develop a methodology to characterise the platelet response phenotype following agonist induced activation, which would then be used to test a cohort of healthy blood donors.

Method  The study consisted of three phases. Phase 1 evaluated a range of platelet function assays in 46 individuals to determine reproducibility and feasibility for a large cohort study. Calcium flux, flow cytometric detection of fibrinogen and Annexin V binding and P‐selectin expression, and aggregometry were used to measure responses to three concentrations of adenosine diphosphate (ADP) and cross linked collagen related peptide (XL‐CRP). From this study, whole blood flow cytometric measurement of fibrinogen binding and P‐selectin expression in response to a single sub‐maximal dose of ADP (10–7 M) and XL‐CRP (0.1 μg mL‐1) were selected for use in phase 2, during which the platelet responses were measured in 506 blood donors attending the Cambridge Centre. In these assays inhibitors of thrombin (hirudin), thromboxane (aspirin), and ADP (apyrase–XL‐CRP stimulated samples only) were included to ensure activation was pathway specific.

Results  There was a wide range of response to both XL‐CRP (fibrinogen binding 3.1%–84.4% +ve; P‐selectin expression 6.2%–90.3% +ve) and ADP (fibrinogen binding 3.5%–78.8% +ve; P‐selectin expression 2.8%–54.1% +ve). Three groups of individuals were selected; two groups with responses to both agonists and both endpoints that were at the ‘Extreme Ends’ of the normal range, (15 low and 16 high responders) and a third group of 16 identified as having mid‐range responses. In phase 3 these 47 donors were re‐tested to confirm the reproducibility of the original classification. There was a high level of concordance between the two visits for the fibrinogen‐CRP, fibrinogen‐ADP, P‐Selectin‐CRP, and P‐Selectin‐ADP assays (P = 0.851, 0.794, 0.855 and 0.527 respectively).

Conclusion  We have demonstrated that different measures of the platelet response to two separate agonists are highly variable between healthy subjects, but are consistent over time within an individual. This standard method for defining platelet functional phenotype is being used in the BLOODOMICS programme to define the association between genetic variation and the risk of atherothrombosis.
SI10 Effects of Hypoxia on the Proliferation and Differentiation Potential of Human Cord Blood‐ and Bone Marrow‐Derived Stem/Progenitor Cells
Authors: Martin‐Rendon E. , Hale S. J. M. , Ryan D. , Baban D. , Forde S. P. , Davies K. , Watt S. M.
Published: September 2006

Umbilical cord blood (UCB) and bone marrow (BM)‐derived stem and progenitor cells possess two characteristics required for successful tissue regeneration, which are extensive proliferative capacity and the ability to differentiate into multiple cell lineages. Within the normal BM and in pathological conditions, areas of hypoxia may have a role in maintaining stem cell fate or determining the fine equilibrium between their proliferation and differentiation. In this study, the transcriptional profiles, proliferation and differentiation potential of UCB CD133+ cells and BM mesenchymal cells (BMMC) exposed to normoxia and hypoxia were analysed and compared. Both progenitor cell populations responded to hypoxic stimuli by stabilising the HIF1‐α protein. Short exposures to hypoxia increased the clonogenic myeloid capacity of UCB CD133+ cells and resulted in a significant increase in BMMC number. The differentiation potential of BMMC and UCB CD133+ cells was unaltered by hypoxia with the lineages tested. When their transcriptional profiles were compared, 183 genes in UCB CD133+ cells and 45 genes in BMMC were differentially regulated by hypoxia. These genes included known hypoxia‐responsive targets such as BNIP3, PGK1, ENO2 and VEGF, and other genes not previously described to be regulated by hypoxia. Several of these genes, namely CDTSPL, CCL20, LSP1,NEDD9, TMEM45A, EDG‐1 and EPHA3 were confirmed to be regulated by hypoxia using quantitative RT‐PCR. These results therefore provide a global view of the signalling and regulatory network that controls oxygen sensing in human adult stem/progenitor cells derived from haematopoietic tissues.
SI11 Endolyn (CD164) Modulates the CXCL12‐Mediated Migration of Umbilical Cord Blood CD133+ and Jurkat T Cells
Authors: Forde S. P. , Jorgensen‐Tye B. , Newey S. E. , Roubelakis M. , Watt S. M.
Published: September 2006

The CXCL12/CXCR4 chemokine axis is a well characterized and important chemotactic stimulus/receptor unit that orchestrates the homing and migration of cells to the bone marrow and to ischemic tissues following tissue damage. Here, we demonstrate that the sialomucin, CD164, a regulator of haemopoietic precursor cell adhesion to stroma and entry of primitive CD34+CD38lo/‐ precursor cells into cycle, modulates the migration of CD133+ cord blood cells to CXCL12 by associating with the CXCR4 receptor. This was demonstrated by a reduction in CD133+ cell migration on fibronectin to CXCL12 (i) by engaging the functional class II glycosylation‐dependent epitope on CD164 with the 103B2/9E10 class II but not the N6B6 class III antibody; and (ii) by RNAi knockdown of CD164 protein levels in CD133+ cells. The inhibition of migration was more pronounced in the more primitive CD34+CD38lo/‐ cell subset. Similar studies using the Jurkat cell line confirmed these findings and led to further analyses using alternative chemokines. A direct association between CXCR4 and CD164 was demonstrated by the co‐localisation of CD164 with CXCR4 and VLA‐4 and VLA‐5 at the leading edge of CD133+ cells when CXCL12 was presented on fibronectin. This was further supported by immunoprecipitation studies that demonstrate in the absence of CXCL12, CXCR4 is associated only with VLA‐4 and VLA‐5 but on exposure to CXCL12, CD164 is rapidly recruited to the CXCR4 complex. Knock‐down of CD164 using siRNA revealed that signalling through CXCR4 via PKC‐ζ was significantly dampened. Our findings therefore support a novel association between three distinct families of cell surface receptors that regulate both cell migratory and proliferative responses and identify a CD164 as a key regulator of the CXCL12/CXCR4 axis.
SI12 Lymphocyte Subsets Following Autologous Transfer of CD25 Depleted Leukapheresis Products
Authors: Austin E. B. , Thistlethwaite F. , Neeson S. , Stern P. , McDonald L. , Hulston M. , Gilham D. , Elkord E. , Griffiths R. , Guest R. , Campbell J. D. M. , Hawkins R. E.
Published: September 2006

A subset of T lymphocytes known as regulatory T‐cells (Tregs) plays a pivotal role in suppressing the immune system in particular the function of effector T‐cells. Tregs have been identified by the cell surface co‐expression of CD4 and high levels of CD25 (IL‐2 receptor α chain) together with CD45RO, CTLA‐4, GITR and the nuclear transcription factor foxp3. Cancer patients with advanced malignancies (renal, gastric etc.) have high numbers of circulating Tregs compared to healthy controls. Experiments in mice indicate a role for Tregs in controlling immune rejection of tumours. In particular, adoptive transfer of CD25+ depleted T‐cell populations induces potent responses to syngeneic tumours. Adoptive transfer of CD25+ depleted autologous Tcells is therefore a potential anti‐cancer treatment. We have begun a study in which autologous leukapheresis products are depleted of CD25+ cells using CD25 CliniMACS® reagent and the CliniMACS® System. The CD25+ depleted product is transferred back to the patient following nonmyeloablative chemotherapy to deplete the host T‐cell repertoire. We report here on the changes to the cellular immune repertoire (CD45, CD3, CD4, CD8, CD19 and CD56) in the first three patients receiving this treatment. In all three cases, the CliniMACS® process gave >2 log depletion of CD25+ lymphocytes. The number of CD3+ cells received were 1.3, 0.6 and 0.9 × 107 kg−1 for patient 1, 2 and 3 respectively. At the time of cell receipt all three patients had CD3 counts of <5 μL−1, rising in patients one and three to above 200 μL−1 by days 9 and 10 respectively, with patient two taking 14–21 days to reach a similar level. The CD4 : 8 ratios were 2.3 and 2.8 (patient 1 and 2, day 7) and 7.3 (patient 3, day 8) dropping to 0.1–0.2 between 14 and 21 days. On day 21 the CD4 : 8 ratio for patient one rose to 1.7 associated with an allergic reaction to prophylactic antibiotic. By day 77 the CD4 : 8 ratios were 0.8 for patient one, and 0.4 for patients two and three. B cell numbers showed signs of recovery in all three patients by day 42. NK cells reached 250–300 μL−1 by days 14–21 but dropped to around 100 μL−1 by day 77 in all three patients. The lymphocyte subset data thus far do not indicate any untoward reactions to the chemotherapy or the depletion of the CD25+ lymphocyte population from the transferred cells.
AW02 Tissues – From Banking to Engineering
Authors: Kearney J. N.
Published: September 2006

Tissue allografts are banked either in the viable or nonviable state. The latter have certain advantages in that the most immunogenic components (e.g. cells and soluble proteins) can be removed from the tissue. This enhances incorporation of the allograft, particularly for tissues with a rapid intrinsic turnover rate (e.g. bone). For other tissues, it has been claimed that the presence of viable donor cells enhances the tissues’ performance in the short to medium term. This is true of skin, where the blood vessels in the skin enosculate with vessels in the wound bed to gain a blood circulation. However this graft is eventually rejected by the recipient. For heart valves, claims of superior performance of viable grafts and long‐term survival of donor cells, are being questioned. The failure of these grafts to re‐cellularise and incorporate has been blamed on immunological responses to immunogenic components. To overcome these problems we have evaluated methods for removing donor cells from soft tissue grafts, without adversely affecting the mechanical and biological properties of the tissue matrix. We have also developed methods for re‐cellularising these tissue matrices with recipients’ cells. For tissues with immediate biomechanical function (e.g. heart valves), bioreactors providing appropriate biomechanical and biological environments have been constructed to simulate in vivo conditions and thus ‘train’ tissue‐engineered grafts, prior to implantation. These techniques are being applied to most of the soft tissue grafts currently supplied by NBS Tissue Services.
SI18 Progress in vCJD Screening for Blood and Cadaveric Donors
Authors: Eglin R.
Published: September 2006

The current position will be discussed regarding the stage of development of the leading blood screening assays for the detection of vCJD and their relative progress towards production of test with the required sensitivity and specificity. The regulatory process to be introduced for the control and standardisation of the use of these assays is in development, with the associated key step of assigning them to Annex II A classification. The application of commercial Western Blot format assays to the detection of PrPsc in tissue samples is underway in NBS and HPA. The validation and implementation of these methods for human tissues demonstrates the paucity of human material available for use as standards and controls. It is appropriate to use animal material for these validation exercises.
SI20 The BBTS Specialist Certificate in Transfusion Science Practice
Authors: Knight R. C.
Published: September 2006

In 1996 the BBTS introduced a certificate in Transfusion Science Practice aimed mainly at staff working in hospital transfusion laboratories who wished to have evidence of their knowledge in this field. Together with the Certificate the BBTS provided a study guide for the distance learning and a textbook, ‘Introduction to Blood Group Serology and Blood Bank Practice’. With recent changes to (state) registration and career grades as a result of Agenda for Change, the Certificate scheme was reviewed and the new Specialist Certificate in Transfusion Science Practice introduced. A new Study Guide has been issued and the latest edition of the introductory book now contains chapters on donation testing and component preparation plus up‐dated sections on immune red cell lysis. The new Specialist Certificate should now meet the requirements of a registered Health Care Scientist working in either a hospital or Blood (Transfusion) Centre wishing to attain the status of a Specialist Practitioner. The examination is very similar to that of the old Certificate except that some choice of questions have been introduced into papers three and four to reflect the wider scope of including donation testing and component production. The practical examination has now been dropped as it was not a good discriminator in terms of the overall examination. With the new ‘donation’ options it would be impossible to set a practical to assess this part of the syllabus. Competency testing has to be the responsibility of the employer. The newly formed Academic Board will be overseeing this examination, its progress and suitability. Any proposed changes would have to be approved by the Professional Affairs and Education Committee. The first of the new examinations was held in May this year and the next will be in November this year and then May 2007.
SI21 Post Registration Training: How Special are You?
Authors: Wainwright A. C.
Published: September 2006

Newly registered practitioners that have completed the Institute's registration portfolio have the opportunity to evidence their development of specialist knowledge and competencies in their chosen field by completion of the Institute's record of laboratory training (Specialist Portfolio). Achievement of the required knowledge and competence of the range of analyses expresses the ability of the biomedical scientist to operate at the specialist level within the Healthcare Scientists career pathway and Agenda for Change band 6. The portfolio will contain a completed and signed record of laboratory training in the designated speciality as evidence of the application of knowledge, comprehension and analytical skills. It will also demonstrate that the individual has reflected on their own performance as an independent professional learner, thus meeting the requirements for CPD. Additional evidence of training and internal assessment of competency will support the Specialist Portfolio; with appropriate cross‐referencing to show which aspect of knowledge or skill acquisition is supported. External verification procedures will establish completion of the portfolio as a record of training in the candidate's ‘home’ discipline, supported by a presentation on the candidate's professional practice and extensive laboratory tour designed to verify the candidate's ability to articulate their scope of practice and ‘fitness for purpose’. The presentation will look at the scope of the portfolio, rationale of the assessment procedures and the use of the portfolio in the context of post‐registration education and training.
PL03 Obtaining Lawful Consent for Donation After Death
Authors: Pearson J.
Published: September 2006

The Human Tissue Act 2004 has afforded communicating with the acutely bereaved about the processes of donation after death is a complex area of clinical practice and as such is undertaken by a team of specially trained nurses. The Human Tissue Act 2004 and the guidance laid down in the Human Tissue Authority Codes of Practice, makes appropriate consent the fundamental principle of the new legislative framework. We see consent as a flow and not an event and this is broken down into key stages which are worked through using conversational style language, which ensures a compassionate delivery of information and a good quality consent or refusal. Within the NBS, the donation conversation is conducted by telephone which has some significant advantages for both the family of the deceased and the organization. The full implementation of the Human Tissue Act in September 2006 brings with it an opportunity to ensure that many more people have their donation wish fulfilled, but there are challenges around this for both NHSBT and the wider NHS.
SI27 Substantial Reductions in Blood Product Usage Achieved Through a Multi‐Discplinary Approach
Authors: Hazlehurst G. , Mold D. , Taylor C.
Published: September 2006

The impact of vCJD on the UK blood supply and the recently well publicised cost pressures on hospital finances make it imperative that hospitals make efforts to reduce the use of blood products. Further drivers for appropriate use of blood and blood products have come from the Department of Health [the two HSCs entitled Better Blood Transfusion (1998 and 2002)] and the National Blood Transfusion Committee [‘National Blood Conservation Strategy for NBTC and NBS’ (2004)]. Objectives can be achieved by a multi‐disciplinary approach with education, ongoing audit and collaboration between the laboratory based staff and all clinical staff. Substantial reduction in the use of blood products can be achieved with the resultant reduction in financial costs and reduction in some of the undesirable effects of transfusion. The Royal Free NHS Trust is a teaching hospital and a tertiary referral centre for haematology, hepatology and nephrology. An extremely active transplantation programme is associated with all three specialities. A multi‐disciplinary approach to blood sparing strategies resulted in a 12.33% reduction (447 K) in blood product contract value. A 9.5% reduction (1680 units) in red cell use was in excess of national averages, greater reductions of 16.8% for FFP (1313 units) and 23.9% (960 units) for platelets were achieved against a background of increased bone marrow transplant activity and greatly expanded renal activity. The substantial reduction in platelet consumption coincided with a more dramatic 79.2% reduction (160 units) in the use of HLA selected platelets for refractory patients. The major contributions to savings involved the close monitoring of blood products, particularly in bone marrow transplant recipients and renal dialysis patients. The increasing use of mini‐allo bone marrow transplants with subsequent rapid recovery of platelet increments, also played a major role in savings. Established protocols for liver transplantation include the use of thromboelastogram (TEG) and aprotinin in selected cases. A pilot study for point of care testing delivered encouraging results. Future initiatives include the expansion of cell salvage to other surgical procedures or haemorrhage scenarios and the use of Erythropoietin (EPO) in some malignancies. Point of care testing is to be expanded to cover more surgical procedures and provide a more accurate assessment of haemostasis closer to real time.
S128 Can the GP Assist in Donor Selection ?
Authors: Vaidyanathan I. , Lee‐Cooper S. , Gorman A.
Published: September 2006

Introduction  To ensure donor and recepient safety when selecting donors we may request additional medical details from the donor's GP or Specialist. This procedure is referred to as GP referral (GPR). The resultant ‘wasted’ session visit and deferral for an unspecified period may adversely affect donor loyalty and deter re‐attendance, while increasing the workload and expense for both NBS and GP surgeries. Furthermore, GPR is perceived by some colleagues to be of questionable value. Because or these important implications we undertook an audit of current practice.

Aims and Objectives  To examine whether GPR is useful, is used appropriately and to identify areas for improving the current practice. The major objectives were to assess whether GPRs were justified, the clinical conditions involved, the relative proportion reflecting donor/recepient safety, the quality of the enquiry, the number and quality of GP replies, the turn‐around time, the outcome of the enquiry and the re‐attendance pattern of reinstated donors.

Method  A total of 242 DHCs with related correspondence were examined retrospectively from three centres first without, then with, the GP reply in order to assess whether GPR was justified. Non justified (avoidable) referrals were defined (Asagba et al 2003) as those in which eligibility could have been determined with the original information given by the donor, and in which the GP reply either endorsed this information or did not alter the outcome.

Results  The majority (90%) of referrals were justified. CVS (43%), NUR (12%) problems generated most referrals. More GPRs reflected donor welfare (69%) than recepient safety. Most of our enquires were focused (98%). Most (96%) doctors replied and 93% of replies were useful. Of 57% reinstated donors, 68% reattended. More regular (83%) than new donors (47%) re‐attended. Average time taken to resolve the enquiry (turnaround time) was 55 days.

Conclusion  GPR makes a significant contribution to the donor selection process. We however recommend further measures to improve the current practice by reducing avoidable GPRs, speedy resolution of enquiry, a donor information leaflet and improved communication with the donors to retain donor loyalty.
SI31 Serious Adverse Events of Blood Donation
Authors: Crusz T. A. M.
Published: September 2006

A single, national system for the identification, recording and follow‐up of serious adverse events associated with blood donation was put in place by the National Blood Service (NBS) in April 2003. As there are no international definitions or categories, we defined a serious adverse event of donation (SAED) as a donor event that results in treatment by or referral to a non NBS clinical professional, e.g. GP, paramedic. Also included in the list of categories is a donor needlestick injury or blood splash contamination to a donor, involving body fluids from a member of staff or another donor. This category was included as these events are not recorded under any other national system. 254 SAEDs (0.21/1000 donors bled) were reported between September 2005 and March 2006. 124 events (48%) were due to vasovagal problems, 80 (31%) were venepuncture related and the rest were due to accidents sustained at session, clinical events directly related to blood donation and medical emergencies which were probably unrelated to donation. 133 events were treated by the GP or practice nurse whilst the remainder were treated in hospital or by paramedics at the site. During this 7 month period 1 needlestick/blood splash contamination event was reported. More up to date figures will be presented. There have been no deaths directly related to blood donation. The NBS has a robust system for not only routinely reporting and recording all donor related adverse events, but also events causing a donor to need clinical care outside the NBS. At present we have no figures from other Blood Services to compare our incidence of SAEDs.
SI36 The Development of a Microarray for Direct Antiglobulin Testing
Authors: Robb J. S. , Knowles L. K. , Petrik J.
Published: September 2006

Background  Blood testing is currently performed using a variety of test platforms. Microarray technology has the potential to include all relevant tests on one testing platform, using the feature of multiplexing. This work demonstrates the development and optimisation of this technology to perform the Direct Antiglobulin Test (DAT) including the examination on the effects of spotting buffer pH on antibody reactivity and the level of sensitisation of DAT positive samples.

Methods  Monoclonal anti‐IgG3 and anti‐C3, rabbit polyclonal anti‐human IgG, monoclonal IgG3 anti‐D plus control proteins were used. This panel was spotted onto a gold microarray surface using specialized microarray preparation equipment and processed under standardised conditions. Directly FITC labelled erythrocytes of Rhesus phenotype R1r were prepared and an aliquot sensitised with the IgG3 anti‐D. Both the native (unsensitised) and sensitised cells were incubated with the microarrays. Following washing, the microarrays were scanned and analysed to determine specifically bound erythrocytes. In a preliminary experiment the polyclonal anti‐IgG was dialysed into pH 7.0, 9.0, 9.8 and 10.4 spotting buffers before spotting. In a subsequent series of experiments, cells were sensitised with varying concentrations of the IgG3 anti‐D, and DAT positive donor blood was included.

Results  The native R1r gave signal/noise (S/N) ratio of up to 100 with IgG3 anti‐D and S/N of <1 with all anti‐IgG and anti‐C3. R1r strongly sensitised with anti‐D IgG3 gave signal/noise (S/N) ratio of over 120 with the anti‐IgGs and <1 with the anti‐D and the anti‐C3.

Conclusions  The microarray demonstrates its suitability for use in the DAT. A high level of sensitisation with IgG blocks the binding of sensitised cells to the same IgG antibody on the microarray. The microarray format can be successfully used for Direct Antiglobulin Testing using a nonagglutination fluorescent readout.
SI39 West of Scotland Blood Centre Audit of Hospital Compliance with Traceability Arrangements for Cross‐Matched Blood
Authors: Mathur K. , Thomson A. , Peterkin M.
Published: September 2006

Introduction  Since 8th November 2005, the Blood Transfusion Services in the UK have been governed by ‘The UK Blood Safety and Quality Regulations 2005’. The Secretary of State for Health, the Competent Authority, can authorise, suspend, or revoke the licence of a Blood Establishment if its activities are not found safe for the purpose of transfusion. The UK Law requires Blood Establishments to establish, and maintain, a quality system based on the Principles of Good Practice (Regulations 6–8). Regulation 8 demands a system for unambiguous vein‐to‐vein traceability and retention and accessibility of traceability data for a minimum period of 30 years. This Blood Centre regularly meets demands to supply cross‐matched blood for patients with multiple irregular allo/auto‐antibodies. To comply with the traceability requirements, we devised a manual system of documentation, a ‘Transfer Receipt’ form, to record the safe supply of cross‐matched blood to Regional hospitals.

Audit Design  Relevant hospital staff were prospectively educated to witness and document the safe and appropriate receipt of blood issues in the bottom section of the accompanying transfer receipt. The completed transfer receipts were returned to the BTC as proof of supply of these components. We audited the completeness of returns of these transfer receipts for a period of 3 months ‐ December 2005–February 2006.

Results  A total of 311 issues of cross‐matched blood were supplied to 21 Regional hospitals during the audit period, with 2/3rd of them (235) involving 10 hospitals, each of which received cross‐matched blood at least once per week. Total returns was 64% (199) and 31% of the returned transfer receipts were incompletely documented. Time of receipt was not recorded on 65% of forms and 21% failed to document the condition of the blood when received. In addition, 16% of receipts were not witnessed and a further 16% were returned with no information. The integrity of blood packs was not documented on a further 3% of transfer receipts. Although completeness of receipts improved during the audit period, the return rate of transfer receipts did not, being 68%, 58%, and 66% in months 1, 2, and 3 respectively.

Conclusion  This audit raises issues about compliance with the UK Blood Safety Law and suggests a need for staff education to ensure that the Law is not breached.
SI04 The Role of the JPAC Standing Advisory Committee on IT (SACIT)
Authors: Penny S.
Published: September 2006

The role of information technology in blood services and hospitals is becoming increasingly important to support the quality of processes and the speed and integrity of information transfer. This holds true from the collection of blood from donors and samples from patients to the processing, testing, distribution and transfusion of blood and blood components. Within this rapidly changing environment the role of SACIT is to provide advice to the UK Blood Transfusion Services. It achieves this by defining standards and recommending these to the UKBTS/NIBSC Joint Professional Advisory Committee (JPAC) for implementation by individual services. To underpin this it defines strategies for how information systems can support the UK services and hospitals, builds relationships with UK and international colleagues working in the field and promotes improvements in Quality through IT. In order to do this the group consists of individuals bringing both expertise in IT and knowledge of the whole ‘transfusion chain’, across the UK, from blood services and hospitals. During 2006/07 the group will work across a range of areas including; driving forward the development of commonality in labelling and bar coding across the UK services, promoting the use of electronic messaging, building relationships with the NHS IT programmes, promoting the further implementation of ISBT 128 and reviewing new technologies.
SI05 Peptide Therapy Suppresses the Adverse Immune Response to Red Cells
Authors: Cairns L. S. , Barker R. N. , Urbaniak S. J.
Published: September 2006

RhD is a clinically important blood group and a major target in haemolytic disease of the newborn (HDN). Current anti‐D antibody therapy to prevent antibody production in RhD negative mothers is ineffective if there has already been sensitization. Peptide immunotherapy offers an alternative approach. Helper T cell epitopes of the RhD antigen were previously identified in alloimmunized donors and we have shown that instillation of these immunodominant RhD‐specific peptides to the nasal mucosa of DR transgenic mice prevents antibody responses to RhD protein. We now wished to test whether such tolerance can be effective in switching off antibody responses in mice that have already been immunized and also the mechanism by which tolerance is achieved. DR15 transgenic mice were immunized and boosted with purified RhD protein, and the immunodominant RhD peptides given nasally, either singly or in a mixture, 4 weeks later. This treatment was found to abrogate helper T cell responses of splenocytes stimulated with RhD protein in culture. This loss in splenocyte activity was often accompanied by an increase in IL‐10 production suggesting the presence of regulatory T cells. In addition, we have demonstrated that antibody levels in the serum declined more quickly in mice that had received the peptides. Thus, nasal instillation of immunodominant peptides provides a method by which to reverse established immune responses to red cells, and this approach may be effective in reducing anti‐D titres in alloimmunized women who are at risk of HDN.
P01 Use of the DiaMed Impact R to Test Platelet Function in Stored Platelet Concentrates
Authors: Morrison A. , Hornsey V. , Macgregor I.
Published: September 2006

Aim  The DiaMed Impact R tests platelet function under close to physiological flow conditions using cone and plate technology. A blood sample is applied to a polystyrene well and plasma proteins (eg VWF and fibrinogen) adhere to the well surface. Shear force is applied to produce arterial flow conditions leading to adhesion and aggregation of platelets via interaction of various platelet membrane glycoproteins including GPIbα and GPIIb/IIIa with vWF and fibrinogen on the well surface. The adhered platelets are quantified by an image analyser and results expressed as % well surface covered by aggregates (SC %) as an index of adhesion and average size of the aggregates (AS μm2) as an index of aggregation. The machine is designed to use whole blood and we found in preliminary studies that adhesion and aggregation do not occur when platelet concentrates (PCs) are used instead. The aim of this study was to add red cells back to PCs so that function could be studied and changes during storage followed and compared with those observed in our established in vitro assays.

Method  ABO compatible routine leucoreduced red cells in SAGM (1 mL) were mixed with 0.6 mL PC to give a final Hct of 35% and platelet count of 400 × 106 mL‐1. These are conditions seen in whole blood except for the reduced levels of leucocytes. Red cells diluted with platelet poor plasma were used as negative controls. The shear rate and time of activation were varied and conditions optimised. Routine PCs were then tested on days 2 and 7. Samples were also stored at 4 °C or 37 °C for 1 to 2 h before assay to see if function could be improved.

Results  Optimum conditions of 1800 s‐1 and 4 min activation produced normal values of SC >7.5% and AS >25 μm2 Mean values ± SD are shown below.

Temperature
Day
SC (%)
AS (μm2)

22°C
2 7
11.1 ± 0.3  5.7 ± 1.4
29.6 ± 0.4 22.2 ± 2.3

4°C
2 7
10.8 ± 1.4  1.9 ± 0.7
26.5 ± 4.1 20.6 ± 1.0

37°C
2 7
14.4 ± 0.1  7.5 ± 0.6
35.8 ± 5.7 24.8 ± 8.6

Conclusions  Red cells are required as well as plasma proteins for platelet adhesion and aggregation to be measurable in the Diamed Impact R. Platelet functions of adhesion and aggregation were shown to decrease during storage but improved at 37°C. The decreases seen were similar to those observed in GPIbα and GPIIb/IIIa binding sites, HSR, ESC and swirl.