Surgical blood order equation in femoral fracture surgery.

AIM: This study aimed at establishing the clinical utility of the surgical blood order equation (SBOE) in patients undergoing femoral fracture surgery. BACKGROUND: A blood ordering schedule defines the perioperative blood use in elective surgery. It lists the number of units of blood required for each procedure preoperatively. MATERIALS AND METHODS: A case-control study was performed among homogeneous groups of patients (n = 62 each) undergoing open reduction and internal fixation of femoral fractures. Correct prediction of blood use in the group of patients using the SBOE was compared to the group whose blood orders were made without any guideline. RESULTS: The surgical blood ordering equation was exactly correct in ordering blood for 46 (74·2%) of 62 patients (cases). The current unaided blood ordering method was exactly correct in ordering blood for 27 (43·5%) of 62 patients (controls). Use of the SBOE resulted in a significantly lower crossmatch-to-transfusion ratio compared to that of the current ordering system (1·5 vs 2·3) and saved the hospital transfusion laboratory 465 US$ of crossmatch and inventory management costs in this cohort of patients. CONCLUSION: The SBOE is a more accurate and cost-saving tool in predicting blood use. It should replace the current unaided method of ordering for perioperative blood in femoral fracture surgery at Mulago Hospital. However, its introduction to other hospitals should be preceded by more rigorous research to strengthen its external validity.

Bottlenecks of blood processing in Uganda.

AIM: To identify where and why delays occur in Uganda blood banks. BACKGROUND: The timely provision and supply of safe and efficacious blood components to hospitals depends on sound systems in the processing blood banks. Poorly managed systems lead to apparent blood shortages in hospitals and increase discard rates due to expiry before dispatch. MATERIALS AND METHODS: We reviewed records of 4126 units of whole blood delivered by the mobile collection teams to a major regional blood bank, in the period 1 March 2009 to 30 June 2009, to ascertain the time intervals between the critical steps in the blood processing chain. This was followed by interviews with staff in two blood banks to establish the causes of process delays. RESULTS: The average duration between blood collection and final labelling (release from quarantine for final storage) was 15·4 (SD 10·8) days. In timeline, the step between matrix generation and grouping was (median duration 8 days) the longest, whereas grouping to labelling was the shortest (median duration 2 days). Blood expiry had the highest discard rate (0·17%) among the non-transfusion transmissible infection marker causes. A minimally facilitated small staff contributed to the process flaws. CONCLUSION: A considerable amount of blood does not reach hospitals because of process delays between collection and ultimate dispatch. This is caused by a thin staff working with inadequate materials, out-of-date methods and in an overcrowded environment. Provision of adequate staff and improved financial allocations to the Uganda Blood Transfusion Services will mitigate this situation.

Informing blood donors about AIDS and risk factors: reactions to information provided in a regional blood bank in the Netherlands.

To safeguard blood transfusion therapy, anti-HIV testing has been standard in routine blood bank practice in the Netherlands since mid-1985. Despite the availability of serological testing for HIV infection, donor deferral remains of prime importance because of the period between HIV-infection and development of antibodies (the 'window phase'). Therefore blood and plasma donors are informed about AIDS and risk factors. Reactions of donors to such information were investigated in a regional blood bank in the Netherlands (1985-1987). Although there seems to be a good understanding of the necessity of reading information about AIDS, personal motives (not belonging to a risk group, having read information before, etc.) resulted in insufficient attention to the written information (a circular letter) by more than one third of respondents. We recommend that information on risk factors is given by a personal approach (i.e. interview) and by emphasizing the personal responsibility of donors for the safety of the blood supply used in blood transfusion therapy.

Intensive chemotherapy with autologous bone marrow transfusion as primary treatment in women with breast cancer and more than five involved axillary lymph nodes.

Patients with breast cancer and a high number of involved axillary lymph nodes have a poor prognosis, despite adjuvant chemotherapy. The 5-year disease-free survival (DFS) in this group amounts to 30-40% and the 10-year DFS is only 15-20%. Therefore, new treatment modalities are being sought for this group of patients. The aim of the present study was the evaluation of the efficacy of high-dose chemotherapy combined with autologous bone marrow support. 24 patients with a primary breast cancer with more than five involved axillary lymph nodes received, after surgery, six courses of induction chemotherapy followed by ablative chemotherapy and reinfusion of autologous bone marrow. All patients were premenopausal or less than 2 years postmenopausal. Induction chemotherapy consisted of methotrexate (MTX) 1.5 g/m2 intravenous (i.v.) and 5-fluorouracil (5-FU) 1.5 g/m2 i.v. on day 1, prednisone 40 mg/m2 orally on days 2-14, doxorubicin 50 mg/m2 i.v. and vincristine 1 mg/m2 i.v. on day 14. Courses were repeated six times every 4 weeks. 10 patients received cyclophosphamide 7 g/m2 i.v. and etoposide 1.5 g/m2 i.v. as intensive regimen, in 14 patients this comprised mitoxantrone 50 mg/m2 i.v. and thiotepa 800 mg/m2 i.v. Reinfusion of autologous marrow followed on day 7. Finally, patients received locoregional radiotherapy for extranodal disease and tamoxifen 40 mg daily orally over a period of 2 years. The median age of patients was 42 years, range 29-54. The median number of involved nodes was 10. During induction therapy, fever requiring i.v. antibiotics occurred in 4% of 144 courses, 14% of patients suffered from mucositis WHO grade 2-3, and the other patients had mucositis grade 1. During the ablative chemotherapy, 1 patient died, 6 developed septicaemia, 5 showed mucositis grade 3-4 and the other patients had mucositis grade 1 or 2. In the follow-up, 1 patient died from acute cardiac failure. Reversible radiation-induced pneumonitis occurred in 7 out of 14 irradiated patients; symptoms started directly following radiotherapy and lasted for several weeks, but disappeared in due course. During follow-up, 2 patients with six and > 10 positive nodes, respectively, have relapsed after 18 and 36 months, both in the cyclophosphamide/etoposide regimen. Median observation is 3 years, disease-free survival at 5 years is predicted to be 84%. Intensive treatment in these patients with high numbers of involved axillary lymph nodes is a toxic regimen, but may improve the chance of surviving free of disease.

Heparin small pool high yield purified factor VIII: in vivo recovery and half-life of routinely produced freeze-dried concentrate.

New approaches and techniques for improving source material collection and Factor VIII production at Blood Bank level have been reported recently. Heparin has been shown to be of importance in increasing yields and stability of FVIII in the purification and concentration process. Work has been done to develop on a routine scale the heparin double cold precipitation technique for the production of a freeze-dried high yield purified FVIII concentrate. The product has been tested clinically in 4 severe hemophilia A patients for recovery, half-life and acute side-effects, using two dosages over 8 infusions. There was no significant difference between the two dosages. Mean recovery 99.1% and mean half-life 8 hr, ranging from 6.5 to 10.3 hr. No side-effects justify further exploration of the potential of heparin for high yield purified FVIII production.

Plasma levels of protein S, protein C, and factor X: effects of sex, hormonal state and age.

We measured total and free protein S (PS), protein C (PC) and factor X (FX) in 393 healthy blood donors to assess differences in relation to sex, hormonal state and age. All measured proteins were lower in women as compared to men, as were levels in premenopausal women as compared to postmenopausal women. Multiple regression analysis showed that both age and subgroup (men, pre- and postmenopausal women) were of significance for the levels of total and free PS and PC, the subgroup effect being caused by the differences between the premenopausal women and the other groups. This indicates a role of sex-hormones, most likely estrogens, in the regulation of levels of pro- and anticoagulant factors under physiologic conditions. These differences should be taken into account in daily clinical practice and may necessitate different normal ranges for men, pre- and postmenopausal women.

Autologous stem cell transplantation in multiple myeloma after VAD and EDAP courses: a high incidence of oligoclonal serum Igs post transplantation.

Thirty-seven patients with multiple myeloma (stage II and III, 65% increased beta2-microglobulin level) were prospectively treated with a median of 3.7 VAD courses (range 2-8) followed by cyclophosphamide (6 g/m2) in conjunction with G-CSF (5 microg/kg filgrastrim (n = 14), or 3.5 microg/kg lenograstrim (n = 22)), and peripheral stem cell (PSC) isolation. After regeneration this was followed by one EDAP course and high-dose melphalan (HDM, 200 mg/m2) in combination with re-infusion of PSC. Adequate stem cell mobilization was obtained with both G-CSF regimens. A median of 41x10(6) CD34+ cells/kg (range 4.5-161) was collected in a median of 1.6 leukapheresis procedures following filgrastrim (n = 14) and 24x10(6) CD34+ cells/kg (range 2. 3-80) in a median of 1.7 leukapheresis procedures following lenograstrim (n = 22) which indicated no significant difference (P = 0.24) between both G-CSF regimens. A rapid hematological recovery was obtained after HDM with reinfusion of a median of 9.3x10(6) CD34+ cells/kg. After the total courses the overall response was 84% with a complete remission rate of 30%. Currently the median overall survival is 44.0 months (95% CI 38.9-49.1) with a median follow-up of 33 months (range 3-51) and a median event-free survival of 29.0 months (95% CI 25.3-32.7) (n = 33). Post transplantation a high incidence of oligloclonal serum immunoglobulins (Igs) was observed. In 73% of the patients new oligoclonal or monoclonal serum bands were noticed 3 months post transplantation. IgG-lambda and IgG-kappa bands predominated. In 48% of the cases the oligoclonal Igs disappeared after a median follow-up of 22 months (range 8-36), whereas in 52% of the cases the oligoclonal Igs persisted with a median follow-up of 31 months (range 21-45), which did not correlate with a significant difference in overall, and event-free survival between both subgroups.

Effects of peripheral stem cell or bone marrow reinfusion on peripheral serotonin metabolism.

Reinfusion of autologous hematopoietic peripheral blood stem cells (PBSC) or bone marrow is often accompanied by flushing, dyspnea, abdominal cramping, nausea and diarrhea. These symptoms and the observation that they can be prevented by ondansetron, a selective 5-HT3 receptor antagonist, led to the assumption that these side-effects are due to infusion of free serotonin during the reinfusion of PBSC or bone marrow. Twenty-five patients with solid tumors received, after myeloblative chemotherapy, a total of 30 reinfusions of PBSC and/or bone marrow. In 17 patients, serotonin levels in the bags containing the PBSC were measured. In all patients, platelet serotonin levels were determined before and 1 h post-reinfusion. In addition, before and 24 h after reinfusion urine was collected for determination of 5-hydroxyindole acetic acid (5-HIAA) and serotonin concentrations. Mean (+/- s.d.) total serotonin concentration in the bags was 2404 +/- 1555 nmol/l. Mean total volume reinfused was 471 +/- 185 ml. After reinfusion, the mean (+/- s.d.) levels of serotonin in platelets in patients increased from 3.2 +/- 1.4 nm/10(9) at baseline to 3.8 +/- 2.0 nm/10(9) (P = 0.02). Neither 24 h urinary 5-HIAA nor serotonin levels were affected. These results indicate that reinfusion of PBSC or bone marrow is accompanied by substantial infusion of free serotonin, which might explain the observed side-effects and justify the use of 5-HT3 receptor antagonists as pre- medication for this procedure.

Platelet aggregability in relation to impaired consciousness after head injury.

ADP-induced platelet aggregation was studied for up to six weeks in 34 patients with head injuries. The patients were divided into three groups according to the degree of impaired consciousness assessed by a clinical coma scale, and change in platelet aggregation was related to the coma score. Platelet aggregation was markedly reduced in all eight patients dying within 24 hours of injury. All 17 patients who remained unconscious for four days or more showed decreased platelet aggregation up to nine days after admission, the most marked effect being on the second day. Platelet function in this group returned to normal within 16 days. Nine patients with only slightly impaired consciousness also showed subnormal platelet aggregation during the first few days with a return to normal by the fourth day. Platelet counts remained within normal limits in all groups. We suggest that during coma following head injury brainstem dysfunction induces neurohumoral changes in the blood which are responsible for a decrease in platelet function.

Factor VIII:C assay: influence of buffer components used in immunoaffinity chromatography for purification of factor VIII/von Willebrand factor.

A monoclonal antibody purified factor VIII concentrate containing FVIII/vWF complex has been assayed by one-stage clotting (CA) and chromogenic substrate (CSA) methods. The influences of potassium iodide (KI) and albumin in combination with predilution buffers, standards and storage of samples have been examined. These components are compared for their effect on FVIII potencies in final product and in-process controls. FVIII:C purified by immunoaffinity chromatography can not be measured reliably by CA or CSA, because of KI which interfere on the assay. Overall yield of FVIII, efficiency of IAC step and purity of FVIII can be determined by assaying the desalted samples.

Comparison of serum S-100 protein levels following stroke and traumatic brain injury.

Temporal changes in serum S-100 protein levels were compared between patients with ischemic stroke, transient ischemic attack (TIA) and traumatic brain injury (TBI). In addition, S-100 levels were correlated with clinical severity and outcome. Measurements were done with a LIA-mat((R)) Sangtec((R)) 100 using an automated immunoluminometric assay. Serum S-100 was measured in 21 stroke patients, 18 TIA patients and ten TBI patients on days 1 (0-24 h), 2, 3, 4, 5 or 6 and 8 or 9. In a control group of 28 healthy volunteers one measurement was done. For the stroke and TIA patients, National Institutes of Health Stroke Scale (NIHSS) scores were obtained on admission and on day 10. For the TBI patients, Glasgow Coma Scale (GCS) scores were obtained on admission and Glasgow Outcome Scale (GOS) scores were obtained after 6 months. Changes in serum S-100 levels over the first 3 days were significantly different between stroke and TBI patients (P=0.014) and between stroke and TIA patients (P=0.006). Peak concentrations of S-100 were most often observed on day 3 or 4 after stroke and on day 1 or 2 after TBI. In the stroke patients individual S-100 peak levels correlated well with the NIHSS score on admission (r=0.58 P=0.014) and the change in NIHSS score between day 10 and day 1 (r=0.65, P=0. 005). In the TBI patients a good correlation between individual peak levels of S-100 and the GCS score on admission (r=-0.81, P=0.010) and the GOS score 6 months after the trauma was found (r=-0.87, P=0. 004). We conclude that there is a significant difference in temporal changes of S-100 levels between ischemic stroke and TBI patients. This suggests different pathophysiological mechanisms. The results of this study further confirm that peak levels of serum S-100 correlate with neurological deficit resulting from either stroke or TBI.

The additive effect of peripheral blood stem cells, harvested with low-dose cyclophosphamide, to autologous bone marrow reinfusion on hematopoietic reconstitution after ablative chemotherapy in breast cancer patients with localized disease.

The additive effect of peripheral blood stem cells (PBSCs) to autologous bone marrow transplantation (ABMT) on haematopoietic reconstitution, after ablative chemotherapy in patients with locally advanced breast cancer, was evaluated. Patients were treated with induction chemotherapy, followed by ablative chemotherapy consisting of mitoxantrone and thiotepa. Group I (n = 14) received ABMT and granulocyte macrophage-colony stimulating factor (GM-CSF), group II (n = 11) received ABMT, PBSCs and granulocyte-colony stimulating factor (G- CSF). PBSCs were harvested after a low-dose cyclophosphamide (750 mg/m2), followed by G-CSF. Stem cell harvest was routinely started 12 days after cyclophosphamide. Compared to group I, group II showed a significant reduction in the median number of days for leukocytes < 0.5 x 10(9)/L 4.5 days, leukocytes < 1.0 x 10(9) / l 5.5 days, platelets < 20 x 10(9)/ l 9 days and platelets < 40 x 10(9) / l 12.5 days. The median number of transfusions of platelets fell from 11.5 to 7 and of red blood cells from 8.5 to 6. The median hospitalisation duration declined from 40.5 to 30 days, fever above 38 degrees C with 7.5 days, fever above 38.5 degrees C with 4 days and antibiotic treatment with 8.5 days in group I versus group II. Improvement of haematological recovery, duration of fever and hospitalisation was observed by the addition of PBSCs, obtained after a relatively low-dose cyclophosphamide and G-CSF and stem cell pheresis on fixed days, to autologous bone marrow and growth factor in the period after ablative chemotherapy.

Recovery of fibrinogen in cryoprecipitate pasteurized in the presence of sucrose and glycine.

The effect of sucrose (60% w/w) and 1 M glycine as thermal stabilizers for fibrinogen in cryoprecipitate was studied. Sucrose (9.2 g) and glycine (0.9 g) were dissolved in 6 g of cryoprecipitate and the solution was pasteurized at 60 degrees C for 10 h. The preparation was then dialyzed for 20 h in phosphate buffered saline (PBS), lyophilized, stored for one week at -40 degrees C and resuspended in distilled water. The recovery of total proteins and fibrinogen in the final product averaged 66.4 +/- 4.1% and 43.8 +/- 6.4% of the initial contents, respectively (mean +/- SEM, N = 9). The pasteurization of cryoprecipitate in the presence of PBS (control experiments) produced extensive precipitation, which is characteristic of protein denaturation. Thus, this method partially protected fibrinogen and other proteins in cryoprecipitate from inactivation by prolonged exposure to heat during pasteurization.

Comparison of apheresis and other methods for separation and purification of hemopoietic stem cells: initial experience with a blood buffy coat model for the use of autologous bone marrow transplantation.

With an increasing number of bone marrow transplantations (BMT) being contemplated in leukemia and cancer patients, it is prudent for blood banks to develop a suitable program within their resources for harvesting, purifying and freezing bone marrow stem cells. In order to do this, initially a prototype has been developed involving buffy coat model (BC) using normal donor blood. Centrifugation, sedimentation and machine apheresis methods were separately evaluated leading to a combined and sequential handling procedure. Blood was passed through a cell separator resulting collection of BC with 90% reduction of the volume showing 80% recovery of total leucocytes and 87% yield of mononuclear cells. Following centrifugation the cells with DMSO were frozen in a controlled freezing system and stored in liquid nitrogen. After thawing 94% cells were recovered with 93% viability. The initial experience gained in the model system could be incorporated in autologous BMT program in patients but requires modifications for improved results; the latter will be described separately.

A simplified method for purification, concentration and freezing of bone marrow cells for autologous transplantation.

Autologous bone marrow (BM) transplantation is being increasingly applied in hematological and oncological patients. However, because of the need to purify and preserve BM requiring high technology, such treatments are virtually concentrated in the "developed" countries. This paper examines methods of BM purification and freezing that could make the technique potentially applicable in developing countries. Hemapheresis is routinely applied for BM purification in our Dutch Centre, where the buffy coats obtained from routine blood donations were utilised in experimental settings. Using DMSO as cryoprotectant, semi-purified white cells were frozen in liquid N2 (LN2), by mechanical freezer or snap-frozen at -55 degrees C. Different types of containers were compared including plastic tubes and ordinary blood bags. After thawing the results show that snap-freezing had a deleterious effect but the cell yields and viability were similar in LN2 or the mechanical freezer where the tubes and the bag were equally effective as containers (86% cell recovery with 90% viability). In the purification/concentration stage, reduction of the volume of the material by extra centrifugation, thus requiring less DMSO, produced better results--96% cell yield and 90% viability after thawing. This simplified method was applied in a general hospital in Sao Paulo where four oncology patients underwent BM collection. BM was purified and concentrated within a blood bank facility. Hydroxyethyl starch sedimentation and centrifugation of the material in plastic blood bags gave 80% BM cell harvest. After thawing from the mechanical freezer 1 x 10(8) BM cells/kg were available for reinfusion to patients. There was no immediate untoward reaction. Three of the patients showed signs of bone marrow regeneration by three weeks, but one patient died 16 days after transplantation, of septicemia. We conclude that certain high-technology procedures for ABMT can be adapted for existing facilities in developing countries.

Plasma exchange in five patients with acute Guillain Barré syndrome.

During the last 3 years plasma exchanges were undertaken in 5 patients with acute Guillain Barré syndrome (G.B.S.). All the patients were admitted in the intensive respiratory care unit and had received six plasma exchange procedures over two weeks (each procedure consists of 2-3 L exchange). The first patient improved dramatically after the second exchange. Moderate success was obtained in two patients. One patient did not show any effect. The fifth patient received plasma exchange one day after her recovery phase had begun but the course of recovery remained uneffected. The effect of plasma exchange was analysed as the patients' response to motor activity, and compared with a historical control group consisting of 50 acute G.B.S. patients admitted in the intensive respiratory care area over the last 25 years. Plasma exchange does not seem to have exerted any significant effect although at any given time the plasma exchange group had higher motor activity than that of the control group. A controlled clinical trial especially in the early phase of the disease is emphasized.

Bone marrow separation, purification and cryopreservation: application for autologous bone marrow transplant (ABMT).

Autologous bone marrow transplantation is used in the treatment of patients suffering from malignant diseases. The intention of this study was to develop a technique for cryopreservation of bone marrow stem cells. A technique of purification described elsewhere (2), gave recoveries of 80% of nucleated blood cells and 87% of mononucleated blood cells after apheresis and centrifugation of buffy-coat, but when applied to bone marrow only 59% of NBC and 32% of CFUc have been harvested. We therefore modified the apheresis technique and replaced the centrifugation step by sedimentation with hydroxy-ethyl-starch. This change resulted in a small but significant increase in the yield of bone marrow cells and improved the asepsis of the procedure. From a second group of bone marrow handled with this modified protocol, final yields of 46% of NBC and 62% of MNC were obtained. After cryopreservation 87% of NBC were recovered as assessed by counting.

Risk management: an important tool for improving quality.

Risk management implies that one has identified and analysed the root cause of the risk. In blood transfusion public and political opinion on the perceived risks are mainly related to product stigmatisation, to cognitive aspects of risk. Therefore this perception is affective, and has a negative connotation. In order to manage risk in an optimal manner, we need to understand how people think about it, and recognise that thoughts, feelings and behaviour are determined not only by psychological factors, but also by social, cultural and political influences. Perception of risk is always situated within a context, which may differ. Therefore people (i.e., public and political opinion) seem to act inconsistently from one risk context to another. Crucial for understanding the logic behind different risk perceptions is how people think about a hazard and organise information about it. The blood supply system has aspects that make it very vulnerable to crises of confidence, as the subject of blood can easily become stigmatised. The impact of the latter on the perception of blood transfusions and their recipients as well as the willingness of the public to accepttransfusions can be dramatic. Risk perception needs to be monitored in order to anticipate and adequately deal with public and political acceptance. We know that risk and stigmatisation are closely interconnected, and that the costs are likely to be high both for human health and for the maintenance of the healthcare system. Thus there is a global need to carefully monitor the safety of the blood supply systems and communicate risk information in a way that both informs people and builds up public and political confidence. It is therefore not sufficient to simply state that the blood supply is safe; it must also be made safe. So risk management becomes an integral part of quality management, as it deals with the public perception of the blood supply system and its respective elements: procurement and use.

[Subspecialty blood transfusion medicine]. / Aandachtsgebied bloedtransfusiegeneeskunde.

The unification of Europe, the related principle of self-sufficiency and the prevention of blood banks turning into bureaucratic institutes that lose connection with bedside medicine underscores the need for training in transfusion medicine and its international organization. In most European countries transfusion medicine is now recognized as a specialty in its own right. In the Netherlands it was decided to recognize transfusion medicine as subspecialty of Internal Medicine. This new initiative led to a training programme of 6 years in all, of which the last 18 months are devoted to transfusion medicine exclusively. The importance of continuous education and practice in both fields is recognized.

[Optimal versus maximal safety of the blood transfusion chain in The Netherlands; results of a conference. College for Blood Transfusion of the Dutch Red Cross]. / Optimale versus maximale veiligheid van de bloedtransfusieketen in Nederland; resultaten van een conferentie. College voor de Bloedtransfusie van het Nederlandse Rode Kruis.

An invitational conference was held on September 11, 1996 by the Medical Advisory Commission to the Blood Transfusion Council of the Netherlands Red Cross, addressing the issues of 'maximal' versus 'optimal' safety measures for the blood supply. Invited were blood transfusion specialists, clinicians, representatives of patient interest groups, the Ministry and Inspectorate of Health and members of parliament. Transfusion experts and clinicians were found to advocate an optimal course, following strategies of evidence-based medicine, cost-benefit analyses and medical technology assessment. Patient groups depending on blood products, such as haemophilia patients would rather opt for maximal safety. Insurance companies would choose likewise, to exclude any risk if possible. Health care juridical advisers would advise to choose for optimal safety, but to reserve funds covering the differences with 'maximal safety' in case of litigation. Politicians and the general public would sooner choose for maximal rather than optimal security. The overall impression persists that however small the statistical risk may be, in the eyes of many it is unacceptable. This view is very stubborn.