Assessment of a closed wash system developed for processing living donor femoral heads.

NHS Blood and Transplant Tissue and Eye Services (TES) and Scottish National Blood Transfusion Services Tissues and Cells Directorate (TCD) currently bank whole, frozen femoral head bone from living donors who are undergoing primary hip replacement surgery. When required, the bone is issued to a surgeon still frozen on dry ice (- 79 °C). Consequently, the femoral head bone is not processed, is not sterilised and at the time of issue, it contains donor blood, bone marrow and associated cells. We have previously shown that, cut, shaped and washed bone from deceased donors can be processed to remove up to 99.9% of blood, bone marrow and associated cells (Eagle et al. 2015). However, cut and shaped bone is not suitable for some orthopaedic procedures and some orthopaedic surgeons do not wish to use irradiated bone; therefore in this report, a method has been developed in which whole femoral heads can be washed to remove donor blood and bone marrow components. Processing results in excess of 99% bone marrow component removal-soluble protein, haemoglobin and DNA; the procedure is performed inside a closed system, thereby eliminating the need for terminal sterilisation by irradiation. In addition, uniaxial testing demonstrated no difference in compressive strength between washed and unwashed bone. We suggest that this washed bone may be capable of improving incorporation after grafting without disturbing biomechanical properties of the graft.

Disuse rescues the age-impaired adaptive response to external loading in mice.

UNLABELLED: We aimed to determine whether aged bone's diminished response to mechanical loading could be rescued by modulating habitual activity. By reducing background loading, aged bone's response to loading increased to a level no different to young mice. This suggests, given the right stimulus, that ageing bone can respond to mechanical loading. INTRODUCTION: Age-related decline in bone mass has been suggested to represent an impaired ability of bone to adapt to its mechanical environment. In young mice, the tibia's response to external mechanical loading has been shown to increase when habitual activity is reduced by sciatic neurectomy. Here we investigate if neurectomy can rescue bone's response to loading in old mice. METHODS: The effect of tibial disuse, induced by unilateral sciatic neurectomy (SN), on the adaptive response to a single peak magnitude of dynamic load-engendered mechanical strain was assessed in 19-month-old (aged) mice. In a second experiment, a range of peak loads was used to assess the load magnitude-related effects of loading on a background of disuse in young adult and aged mice. Bone architecture was analysed using micro-computed tomography (µCT) and dynamic histomorphometry. RESULTS: In the first experiment, SN in aged mice was associated with a significant periosteal osteogenic response to loading not observed in sham-operated mice (7.98 ± 1.7 vs 1.02 ± 2.2 % increase in periosteally enclosed area, p < 0.05). In the second experiment, SN abrogated the expected age-related difference in the bones' osteogenic response to peak strain magnitude (p > 0.05). CONCLUSIONS: These data suggest that bones' age-related decline in osteogenic responsiveness to loading does not originate in bone cells to either assess, or appropriately respond to strain, but rather is likely to be due to inhibitory "averaging" effects derived from the habitual strains to which the bone is already adapted. If such "strain averaging" is applicable to humans, it suggests that gentle exercise may degrade the beneficially osteogenic effects of short periods of more vigorous activity.

The cyclooxygenase-2 selective inhibitor NS-398 does not influence trabecular or cortical bone gain resulting from repeated mechanical loading in female mice.

UNLABELLED: A single injection of the cyclooxygenase-2 (COX-2) selective inhibitor NS-398 reduces bone's osteogenic response to a single period of mechanical loading in female rats, while women taking COX-2 selective inhibitors do not have lower bone mass. We show that daily NS-398 injection does not influence bone gain from repeated loading in female mice. INTRODUCTION: Prostaglandins are mediators of bone cells' early response to mechanical stimulation. COX-2 expression is up-regulated by exposure of these cells to mechanical strain or fluid flow, and the osteogenic response to a single loading period is reduced by COX-2 inhibition. This study determined, in female mice in vivo, the effect of longer term COX-2 inhibition on adaptive (re)modelling of cortical and trabecular bone in response to repeated loading. METHODS: Nineteen-week-old female C57BL/6 mice were injected with vehicle or NS-398 (5 mg/kg/day) 5 days a week for 2 weeks. On three alternate days each week, the right tibiae/fibulae were axially loaded [40 cycles (7 min)/day] three hours after injection. Left limbs acted as internal controls. Changes in three-dimensional bone architecture were analysed by high-resolution micro-computed tomography. RESULTS: In control limbs NS-398 was associated with reduced trabecular number but had no influence on cortical bone. In loaded limbs trabecular thickness and cortical periosteally enclosed volume increased. NS-398 showed no effect on this response. CONCLUSION: Pharmacological inhibition of COX-2 by NS-398 does not affect trabecular or cortical bone's response to repeated mechanical loading in female mice and thus would not be expected to impair the functional adaptation of bone to physical activity in women.

Collection, processing and testing of bone, corneas, umbilical cord blood and haematopoietic stem cells by European Blood Alliance members.

BACKGROUND AND OBJECTIVES: A questionnaire study was carried out in collaboration with the European Blood Alliance (EBA) Tissues and Cells (T&C) working group. The aim was to assess the level of involvement and commonality of processes on the procurement, testing and storage of bone, corneas, umbilical cord blood (UCB) and haematopoietic stem cells (HSC) in order to identify different practices and to explore whether recommendations can be made for harmonization. MATERIALS AND METHODS: An online questionnaire was used for data collection in 2011, and 43 replies were received covering 71 product answers from 13 countries. RESULTS AND CONCLUSIONS: Estimated percentages of tissue and cell banking covered by EBA member blood banks as a proportion of all collections of each individual country varied markedly. There were also major differences in the amounts of products collected and discarded and in proportions tissues provided for grafting. However, discarding of certain collections also reflects the practice of increasing the likelihood of the very best units being used for transplantation. Harmonization of possible practices should focus on matching supply with demand and on identifying the most efficient operators. This could allow for the development of practices for minimizing unnecessary collections.

Prevention and control of childhood asthma and allergy in the EU from the public health point of view: Polish Presidency of the European Union.

The leading priority for the Polish Presidency of the Council of the European Union was to reduce health inequalities across European societies, and, within its framework, prevention and control of respiratory diseases in children. This very important paper contain proposal of international cooperation on the prevention, early detection and monitoring of asthma and allergic diseases in childhood which will be undertaken by the EU member countries as a result of EU conclusion developed during the Polish Presidency of the Council of the European Union. This will result in collaboration in the field of chronic diseases, particularly respiratory diseases, together with the activity of the network of national institutions and NGOs in this area. Paper also contains extensive analysis of the socio-economic, political, epidemiological, technological and medical factors affecting the prevention and control of childhood asthma and allergy presented during Experts presidential conference organized in Warsaw-Ossa 21-22 September 2011.

Intra-operative washing of morcellised bone allograft with pulse lavage: how effective is it in reducing blood and marrow content?

The use of unprocessed bone carries a risk of transmission of blood borne diseases. Although models of infectivity are unproven, a theoretical risk of transmission of variant Creutzfeld-Jakob Disease, a human prion disease, exists as probable blood borne transmission has been reported in three cases. The aim of our study was to determine the effectiveness of standard operating theatre pulse lavage in removing protein, fat and double stranded Deoxyribonucleic acid (dsDNA) from morcellised bone allograft. Twelve donated femoral heads were divided into halves and milled into bone chips. One half of the bone chips were washed with pulse lavage, whereas, the other half acted as control. In order to determine the amount of protein, fat and dsDNA present in the washed and unwashed samples, a validated multistep washing protocol was used. Using the validated technique, simple intra-operative washing of morcellised unprocessed bone allograft removed a significant amount of the protein (70.5%, range: 39.5-85%), fat (95.2%, range: 87.8-98.8%) and DNA (68.4%, range: 31.4-93.1%) content. Intra-operative washing of morcellised bone allograft with pulse lavage may thereby reduce the theoretical risk of prion and other blood borne disease transmission. Combined with the known improved mechanical characteristics of washed allograft, we would recommend pulse lavage as a routine part of bone allograft preparation.

The organization of tissue banking in Scotland.

Tissue banking in Scotland has developed significantly over the past 20 years or so. The range of issues procured has increased and so have their numbers. Initially, bone from live donors was the only tissue banked; later, tissues from multiorgan donors were procured; this was finally followed by the collection of tissues from donors following cardiac death. Bones, tendons, heart valves and skin are the main tissue types collected, stored and issued for clinical use. Much of our activity is based on identification of donors in two major accident and emergency departments followed by retrievals that take place in a dedicated mortuary by fully-trained staff. Tissues are released according to clinical need for Scottish patients and beyond. All of the tissue banking activity in Scotland takes place within the Scottish National Blood Transfusion Service, which is the preferred provider of tissues for Scottish patients. There is very close cooperation between our teams, the transplant teams and other clinical colleagues, including pathologists and anatomical technicians. The achievements in issue banking in Scotland are outlined along with the main clinical indications of the tissue procured. Diversification is now taking place into cellular therapy with the establishment of an islet processing programme and cell culturing techniques. The future is very exciting.

Estimating the potential of tissue donation in Scotland.

The Scottish National Blood Transfusion Service became the preferred provider of tissues in 2000. At that time, the only tissues procured were heart valves and some tendons from multiorgan donors. These sources and the range of tissues were not enough to provide for the clinical needs of Scottish patients. A systematic review was undertaken using the International Classification of Disease codes to assess what the potential pool of tissue donors would be by region, hospital and hospital department. Such data would enable us to focus our limited resources to maximize the yield of tissues to meet clinical demand. The data from this study were validated by a case-note audit in one hospital in Edinburgh. It was shown that the maximum tissue donor potential of approximately 48.5 per million population was in the central belt of Scotland in hospitals with large emergency medicine departments.

Hygiene behaviour of adolescents in the Pacific: associations with socio-demographic, health behaviour and school environment.

This study presents findings on health-related hygiene behaviours of 9,013 adolescents from Vanuatu, Tonga and Pohnpei in the Federal States of Micronesia as part of the Health Behaviour of Pacific Youth Life surveys. We examined the prevalence of and relationship between tooth brushing, hand washing before eating, hand washing after toileting and a range of psychosocial factors such as sociodemographic charac-teristics, health behaviour and school affiliation. The results showed that patterns of prevalence and relationships were consistent for all countries, with weaker associations observed in Pohnpei. In general, adolescents reported moderate levels of optimal hygiene practice. Girls reported significantly more frequent optimal hygiene practice than boys. Logistic regression analysis confirmed that frequent hygiene behaviours were significantly associated with gender, parental occupations and high levels of school affiliation. These findings suggest the need to consider psycho-social and economic factors when examining potential influences on hygiene behaviour to ensure effective interventions.

Accurate and semi-automated analysis of bacterial association with mammalian cells.

To efficiently and accurately quantify the interactions of bacteria with mammalian cells, a reliable fluorescence microscopy assay was developed. Bacteria were engineered to become rapidly and stably fluorescent using Green Fluorescent Protein (GFP) expressed from an inducible Tet promoter. Upon application of the fluorescent bacteria onto a monolayer, extracellular bacteria could be discriminated from intracellular bacteria by antibody staining and microscopy. All bacteria could be detected by GFP expression. External bacteria stained orange, whereas internalised bacteria did not. Internalised bacteria could thus be discriminated from external bacteria by virtue of being green but not orange fluorescent. Image acquisition and counting of various fluorophore-stained entities were accomplished with a high-content screening platform. This allowed for semi-automated and accurate counting of intracellular and extracellular bacteria.

Global priority setting for Cochrane systematic reviews of health promotion and public health research.

BACKGROUND: Systematic reviews of health promotion and public health interventions are increasingly being conducted to assist public policy decision making. Many intra-country initiatives have been established to conduct systematic reviews in their relevant public health areas. The Cochrane Collaboration, an international organisation established to conduct and publish systematic reviews of healthcare interventions, is committed to high quality reviews that are regularly updated, published electronically, and meeting the needs of the consumers. AIMS: To identify global priorities for Cochrane systematic reviews of public health topics. METHODS: Systematic reviews of public health interventions were identified and mapped against global health risks. Global health organisations were engaged and nominated policy-urgent titles, evidence based selection criteria were applied to set priorities. RESULTS: 26 priority systematic review titles were identified, addressing interventions such as community building activities, pre-natal and early infancy psychosocial outcomes, and improving the nutrition status of refugee and displaced populations. DISCUSSION: The 26 priority titles provide an opportunity for potential reviewers and indeed, the Cochrane Collaboration as a whole, to address the previously unmet needs of global health policy and research agencies.

Haematological and haemorheological changes associated with cigarette smoking.

A prospective study was undertaken to establish the haematological and haemorheological changes associated with long standing cigarette smoking in 20 heavy smokers and to assess whether any such changes were reversible after smoking was stopped. Highly significant differences were observed in whole blood viscosity, plasma viscosity, plasma fibrinogen concentrations, packed cell volume, and carboxyhaemoglobin concentrations between smokers and non-smokers. Ten of the subjects were followed up for two weeks after stopping smoking by which time whole blood viscosity and carboxyhaemoglobin concentrations had improved significantly and indications of improvement could be seen in all other measurements. Two of these subjects were further followed up for two months when all the measured variables were comparable with those in non-smokers. No correlation could be established between carboxyhaemoglobin concentration and any of the other variables. Although patients' compliance may be difficult to obtain, further prospective studies would be required to confirm our findings.

Supply and demand of bone allograft for revision hip surgery in Scotland.

The Scottish National Blood Transfusion Service is the main provider of bone for grafting in Scotland. Bone is procured only from live donors, following very strict selection criteria, and we have investigated whether the amount being collected was adequate. Our current harvest of approximately 1700 femoral heads per year is shown not to be enough to meet the future demand for revision surgery of the hip. Many more of these operations are being undertaken, and impaction grafting is being used increasingly. We have calculated the predicted rates of collection and usage for the next four to five years so that we can expand our service in a controlled fashion.

A simplified pharmacologic erection program for patients with spinal cord injury.

INTRODUCTION AND OBJECTIVES: Intracavernosal injection (ICI) of vasoactive agents has been successfully used in the treatment of erectile dysfunction (ED). The authors' pharmacologic erection program, using a fixed combination of prostaglandin E1 (PGE1) and papaverine, is notable for its simplicity and acceptance by patients who have suffered spinal cord injuries (SCIs). METHODS: Patients undergo baseline questionnaire, physical examination, and hormone profile followed by instruction and injection of a fixed combination of PGE1 and papaverine. On successive visits, the patient injects himself and the dosage is titrated until a satisfactory erection is obtained. Patients who do not respond to injection of 1.0 cc are considered treatment failures. Patients return periodically for routine follow-up. RESULTS: From May 1994 to March 1997, 37 patients with SCI underwent initial evaluation and 28 (76%) responded to injection therapy and were successfully using self-injection therapy at 3-month follow-up. Twenty-three patients are still on injection therapy. Five patients have dropped out for several reasons including a lack of a current sexual partner (60%) and pain with injection (40%). Patient age ranges from 24 to 72. The dosage range was 0.10 to 0.50 cc (mean = 0.29 cc). The average duration of erection was 43 minutes. At 3-month follow-up, 85% of the patients rated their erections as good or excellent. Forty-three percent of patients are using ICI 1 or more times per week. Seventy-seven percent of patients are moderately or extremely satisfied with their treatment and 89% said that they would recommend this program to a friend. CONCLUSIONS: This simplified pharmacologic erection program offers safe, well accepted, and effective therapy for ED to a SCI population with very high patient satisfaction.

Efficacy and safety of oral sildenafil in men with erectile dysfunction and spinal cord injury.

OBJECTIVE: To assess the efficacy of sildenafil in men with spinal cord injury (SCI) and erectile dysfunction (ED). METHODS: Seventeen men with SCI were selected from February to September 1998 for sildenafil treatment of ED. The initial dose of 25 mg was increased by 25-mg increments as needed. Patients underwent baseline physical examination and answered questions from the abridged International Index of Erectile Function before and during therapy. RESULTS: Sixteen patients tolerated therapy; 1 developed hypotension and discontinued therapy. There was significant improvement in erectile function (P < .05) after 5.3 +/- 2.2 months when compared with baseline or previous therapies (P < .05). Of the 17 patients, 94% recommended sildenafil to others. Six of these 16 patients were available for long-term follow-up. There was further significant improvement in quality of erection (P < .05), but no change in satisfaction. CONCLUSION: Sildenafil is effective and well tolerated in men with SCI and ED.

Utilisation of bone allograft by orthopaedic surgeons in Scotland.

The use of bone allograft in orthopaedic surgery has been predicted to increase, particularly in joint revision surgery. This has led to a potential problem with supply. Questionnaires were distributed to all 146 Consultant Orthopaedic surgeons working in Scotland in 2000. They were asked to indicate their current usage of bone and tissue allograft, any problems encountered with supply and if alternatives to allograft, such as processed bone, might be used. The questions asked were very similar to those asked in a previous study in 1995 to enable comparisons to be made. Replies were received from 125 Consultants (87%) of whom 93 reported using bone allograft. Forty-one consultants (46%) predicted an increase in their requirement for bone allograft, and 23 (26%) felt they could currently use more bone if this was available. Sixty percent of surgeons would consider using processed bone as an alternative. In comparison with figures from 1995, an increasing number of surgeons are prepared to use processed bone as an alternative to fresh frozen allograft.

Estrogen receptors α and ß have different gender-dependent effects on the adaptive responses to load bearing in cancellous and cortical bone.

To determine the effect of estrogen receptors (ER) α and ß on bones' adaptive response to loading, we subjected the right tibiae of mice lacking ERα or ERß activity to either axial loading or to disuse. Adaptive changes in architecture were assessed by comparing differences between the right (treated) and left (control) tibiae in these genotypes as assessed by microcomputed tomography. In female ERα(-/-) mice, the net-osteogenic response to loading was lower in cortical bone compared with their wild-type littermates (11.2 vs. 20.9% in ERα(+/+)), but it was higher in both cortical and cancellous bone of male ERα(-/-) mice (cortical 20.0 vs. 4.6% in ERα(+/+); cancellous 30.0 vs. 5.3% in ERα(+/+), P < 0.05). In ERß(-/-) male and female mice, the net-osteogenic response to loading was higher in cortical bone (males 10.9 vs. 3.9% in ERß(+/+); females 18.5 vs. 15.8% in ERß(+/+), P < 0.05) but no different from controls in cancellous bone. The bone loss in response to disuse was less in cancellous bone of ERα(-/-) mice than in controls (-15.9 vs. -21.3%, respectively, P < 0.05) but no different at any other site or between any other groups. Our conclusion is that functional ERα enhances the net-osteogenic response to loading in cortical but not cancellous bone in female mice but reduces it in males. ERß decreases the response to loading in cortical bone of males and females but has no effect in cancellous bone. Bone loss due to disuse in cortical bone is unaffected by ER status, but in cancellous bone, functional ERα contributes to greater disuse-related bone loss.