Long chain polyunsaturated fatty acid supplementation in infancy increases length- and weight-for-age but not BMI to 6 years when controlling for effects of maternal smoking.

Long chain polyunsaturated fatty acids (LCPUFA) are added to infant formula but their effect on long-term growth of children is under studied. We evaluated the effects of feeding LCPUFA-supplemented formula (n = 54) compared to control formula (n = 15) throughout infancy on growth from birth-6 years. Growth was described using separate models developed with the MIXED procedure of SAS(®) that included maternal smoking history and gender. Compared to children fed control formula, children who consumed LCPUFA supplemented formula had higher length-/stature-/and weight-for-age percentiles but not body mass index (BMI) percentile from birth to 6 years. Maternal smoking predicted lower stature (2-6 years), higher weight-for-length (birth-18 months) and BMI percentile (2-6 years) independent of LCPUFA effects. Gender interacted with the effect of LCPUFA on stature, and the relationship between smoking and BMI, with a larger effect for boys. Energy intake did not explain growth differences. A relatively small control sample is a limitation.

Cognitive analysis of decision support for antibiotic ordering in a neonatal intensive care unit.

BACKGROUND: Clinical decision support systems (CDSS) are a method used to support prescribing accuracy when deployed within a computerized provider order entry system (CPOE). Divergence from using CDSS is exemplified by high alert override rates. Excessive cognitive load imposed by the CDSS may help to explain such high rates. OBJECTIVES: The aim of this study was to describe the cognitive impact of a CPOE-integrated CDSS by categorizing system use problems according to the type of mental processing required to resolve them. METHODS: A qualitative, descriptive design was used employing two methods; a cognitive walkthrough and a think-aloud protocol. Data analysis was guided by Norman's Theory of Action and a theory of cognitive distances which is an extension to Norman's theory. RESULTS: The most frequently occurring source of excess cognitive effort was poor information timing. Information presented by the CDSS was often presented after clinicians required the information for decision making. Additional sources of effort included use of language that was not clear to the user, vague icons, and lack of cues to guide users through tasks. CONCLUSIONS: Lack of coordination between clinician's task-related thought processes and those presented by a CDSS results in excessive cognitive work required to use the system. This can lead to alert overrides and user errors. Close attention to user's cognitive processes as they carry out clinical tasks prior to CDSS development may provide key information for system design that supports clinical tasks and reduces cognitive effort.

An evaluation of the range and availability of intensive smoking cessation services in Ireland.

BACKGROUND: A review of smoking cessation (SC) services in Ireland is a necessary step in improving service planning and provision. AIMS: To assess the range and availability of intensive SC services in Ireland in 2006. METHODS: A survey of SC service providers in Ireland was conducted. Descriptive analysis and simple linear regression analysis was used. RESULTS: Response rate was 86.3% (63/73). All service providers surveyed are employing evidence-based interventions; the most common form of support is individual counselling with initial sessions averaging 40 min and weekly review sessions 20 min in duration. Reaching the recommended target of treating 5.0% of smokers does not seem feasible given the current distribution of resources and there appears to be regional differences in resource allocation. CONCLUSIONS: While intensive SC services are available in all four Health Service Executive Areas, it would appear that there is little uniformity or consistency countrywide in the scope and structure of these services.

Smoking profile among the gay and lesbian community in Ireland.

OBJECTIVE: We hypothesized that smoking rates among the Gay and Lesbian Community (GLC) in Ireland are not significantly different from the general Irish population. METHODS: A convenience sampling of self-identified GLC was recruited using electronic (n = 700) and print (n = 500) media procedures in response to survey call advertisements (December 2006-March 2007). In all, 1,113 had complete smoking data and were analyzed. Data on a random sample of 4,000 individuals, using the Irish Office of Tobacco Control monthly telephone survey, were analyzed for the same period. RESULTS: Adjusted smoking rates in GLC were 26 and 24.6% in the general Irish population (P = 0.99), while "heavy" (> or =20 cigarettes/day) smoking prevalence was 44.1 and 36.6%, respectively (P = 0.02). Upper SES GLCs are "heavy" smokers compared with general population of similar SES group (P = 0.01). CONCLUSION: When considering two different sampling methodologies, this study suggests that smoking rates among the GLC in Ireland are not significantly different from the general Irish population.

Prevention of monocyte adhesion and inflammatory cytokine production during blood platelet storage: an in vitro model with implications for transfusion practice.

A novel platelet additive solution [ThromboSoltrade mark (TS)] was designed to allow extended refrigerated platelet storage. It has been shown to preserve platelet function and prevent cytokine accumulation in platelet concentrates stored for up to 9 days. It consists of amiloride, adenosine, sodium nitroprusside, dipyridamole, quinacrine, and ticlopidine. We hypothesized that the cytokine inhibition may be due to prevention of monocyte (MC) adhesion and activation on the surfaces of platelet storage bag plastic polymers. In an in vitro model, we incubated purified peripheral blood MCs on discs of polyolefin and polyvinylchloride from platelet storage bags, and on polystyrene, in the presence of TS for up to 7 days. We found that after incubation with TS, adherent MC numbers were decreased by >80-95% compared with controls on all surfaces examined. Levels of cytokines [interleukin (IL)-1beta, IL-1RA, IL-6, IL-8, and tumor necrosis factor-alpha] were low in wells with TS but rose progressively in the controls during incubation. Amiloride alone had similar effects on adhesion and cytokine release as the complete TS preparation. Removing amiloride from TS abrogated these effects. These findings suggest an important role for TS and amiloride in monocyte function, and have implications for the development of agents designed for prolonged platelet storage.

Enhanced circulatory parameters of human platelets cryopreserved with second-messenger effectors: an in vivo study of 16 volunteer platelet donors.

Platelet transfusion represents an important component of the therapy for thrombocytopenic patients. Prolonged storage capabilities for platelets would alleviate many problems associated with blood banking. Unfortunately, current cryopreservation methods are complex to implement and result in loss of cell number and functional activity. Previous in vitro studies have shown that the use of ThromboSolTM, a platelet-stabilizing formulation, in the cryopreservation of platelets results in significant retention of cell number and in vitro functional activities in addition to reducing the DMSO requirement to only 2%. We evaluated the in vivo circulatory parameters of platelets cryopreserved with ThromboSol. Single donor platelet units were obtained from healthy volunteers (n = 16); the units were then split and cryopreserved with either ThromboSol and 2% DMSO or 6% DMSO alone. Following storage at -80 degrees C for 7-10 d the samples were thawed, washed and radiolabelled with either 51Cr or 111In. The paired samples were then mixed and reinfused into the autologous volunteer. At various time intervals following transfusion a blood sample was drawn and the quantity of circulating labelled platelets was determined. The percent recovery and survival time was determined by multiple-hit analysis. The ThromboSol-treated platelets, as compared to the 6% DMSO-treated platelets, displayed statistically higher percent recovery (40.2% v 28.8%) and survival time (166.3 h v 152.1 h). These results demonstrated that platelets cryopreserved with ThromboSol displayed superior in vitro and in vivo characteristics as compared to the standard 6% DMSO method. The use of ThromboSol allowed for a 3-fold reduction in the DMSO concentration in conjunction with a 40% increase in circulating cell number and normal survival times.

Enhanced retention of in vitro functional activity of platelets from recombinant human thrombopoietin-treated patients following long-term cryopreservation with a platelet-preserving solution (ThromboSol) and 2% DMSO.

Chemotherapy-induced thrombocytopenia represents a significant clinical problem in the management of patients with malignancy. Recombinant human thrombopoietin (rhTPO) is a potent stimulator of platelet production in vivo. The ability to cryopreserve rhTPO-derived platelets would enable the use of autologous platelets during the period of thrombocytopenia. ThromboSol is a platelet-stabilizing formulation consisting of second messenger effectors that inhibit specific activation pathways endogenous to platelets. To investigate the effect of ThromboSol cryopreservation, platelets from rhTPO-treated patients (n = 23) and normal donors were treated with ThromboSol and 2% DMSO and cryopreserved for up to 6 months. The platelets were thawed at different intervals and tested for retention of platelet functional activity in vitro. Following a short-term storage (1 week), the cryopreserved platelets from patients treated with rhTPO exhibited significantly higher retention of functional activities including discoid morphology (70% v 57%), extent of shape change (19% v 13%) stirring shape change (15% v 11%) and hypotonic shock response (56% v 25%), as compared to the cryopreserved platelets from controls. Furthermore, there was no further significant loss of functional activity following cryopreservation for up to 6 months. These findings suggest that cryopreservation of platelets from rhTPO-treated donors may provide a useful novel strategy for autologous or allogeneic donation for subsequent transfusions to manage treatment-related thrombocytopenia.

Cryopreservation of single-donor platelets with a reduced dimethyl sulfoxide concentration by the addition of second-messenger effectors: enhanced retention of in vitro functional activity.

BACKGROUND: The potential for bacterial contamination limits the storage of platelets at 22 degrees C to 5 days. This creates an inventory problem, which could be overcome by the use of cryopreservation to allow long-term storage of platelets. It has been demonstrated that the addition to platelets of a mixture of second-messenger effectors (platelet storage solution), allows these cells to retain significant in vitro functional activity following cold storage. Analysis is needed of the ability of this second messenger effector mixture both to protect platelets during cryopreservation and to reduce the need for a cryoprotectant. STUDY DESIGN AND METHODS: Fresh single-donor platelet units (n = 8) were divided into three samples and treated with 6-percent dimethyl sulfoxide (DMSO), 2-percent DMSO or the platelet storage solution and 2-percent DMSO. The samples were placed directly into a -80 degrees C freezer and stored for 1 week, after which they were thawed and analyzed for in vitro functional activity. RESULTS: Platelets cryopreserved with the platelet storage solution and 2-percent DMSO displayed statistically higher retention of functional activity and viability--including cell number, percent of discoid cells, extent of shape change, and hypotonic shock response--than did platelets stored by the method using 6-percent DMSO. In addition, the treated platelets displayed statistically lower expression of p-selectin. The treated platelets showed no loss of cell number, > 88-percent retention of discoid morphology, and > 75-percent retention of ristocetin-induced aggregation as compared to values for these measures in fresh platelets. CONCLUSION: The use of this platelet storage solution in the cryopreservation of platelets yields a significant improvement in their postthaw in vitro recovery and allows for a reduction of the DMSO concentration from 6 to 2 percent, with superior maintenance of in vitro viability and function.

Inhibition of cytokine accumulation and bacterial growth during storage of platelet concentrates at 4 degrees C with retention of in vitro functional activity.

BACKGROUND: The potential for bacterial contamination limits the storage of platelet concentrates (PCs) at 22 degrees C to 5 days. In addition, storage of platelets under conventional protocols for longer times (> 3 days), in the absence of white cell filtration, has been correlated with incidents of cytokine-associated febrile reaction in recipients. It has been demonstrated that the addition of a reagent mixture of second-messenger effectors allows platelets stored at 4 degrees C to maintain significant in vitro functional activity. Thus, the effects of 4 degrees C storage on the growth of bacteria and the accumulation of cytokines by the white cell fraction of PCs were analyzed to demonstrate the benefits of this refrigerated storage system. STUDY DESIGN AND METHODS: The platelet storage solution was added directly to PCs obtained from the blood bank, and these treated PCs were stored at 4 degrees C without agitation. In parallel, control PCs were stored according to standard blood-banking procedures. On Days 1, 3, 5, and 9, the PCs were measured for the plasma concentrations of cytokines. Treated and control PCs stored at 4 degrees C and 22 degrees C were inoculated with low-titer Staphylococcus aureus, and bacterial growth was measured over a 5-day period. RESULTS: Control PCs displayed a time-dependent increase in the plasma concentration of interleukin 6, interleukin 1 beta, and tumor necrosis factor alpha. These conventionally stored PCs also displayed a time-dependent increase in the bacteria titer. In contrast, the treated PCs stored at 4 degrees C displayed no accumulation of the above cytokines in the plasma fraction and no increase in bacteria titer above the initial inoculation. CONCLUSION: The storage of PCs at refrigerated temperatures inhibits the accumulation of white cell-produced cytokines in the PCs, an effect that could alleviate cytokine-associated febrile transfusion reactions The 4 degrees C storage was also bacteriostatic, which indicates that the storage of PCs at that temperature increases safety by decreasing the potential for sepsis. Thus, the ability to store PCs at 4 degrees C may allow extension of the storage limit beyond 5 days.

Recovery of in vitro functional activity of platelet concentrates stored at 4 degrees C and treated with second-messenger effectors.

BACKGROUND: The potential for bacterial contamination limits the storage of platelets at 22 degrees C to 5 days. Refrigerated storage at 4 degrees C would abrogate this problem but would also result in a rapid loss of in vitro viability and functional activity and in vivo viability. The inhibition of platelets during storage by a combination of specific, reversible, second-messenger effectors has been investigated to allow prolonged storage at 4 degrees C with significant retention of in vitro viability and functional activity. STUDY DESIGN AND METHODS: The combination of effectors was added directly to platelet concentrates, and this step was followed by storage at 4 degrees C. Control units were incubated at 4 degrees C without the effectors and at 22 degrees C according to standard blood-banking techniques. At 1, 5, and 9 days, the units were tested for recovery of cell number, recovery of in vitro functional activity and viability, and expression of platelet surface markers. RESULTS: Treated platelets stored at 4 degrees C for 9 days, while spherical in shape, displayed no loss of cell number and had a recovery of viability and functional activity, as compared with control platelets stored at 22 degrees C for 5 days, as follows: ADP and collagen aggregation responses of 250 and 100 percent, respectively; a 70-percent recovery of hypotonic shock response; and a 60-percent recovery of extent of shape change. The treated platelets also expressed an equivalent amount of the surface marker glycoprotein lb and a lower amount of the activation marker alpha-granule membrane protein-140 on the membrane surface. CONCLUSION: Second-messenger effectors added to platelets significantly maintained in vitro functional activity with storage at 4 degrees C. In vitro analysis demonstrates the potential for extended 4 degrees C storage of platelets with numerical and functional recovery comparable to that achieved with current methods. Refrigerated storage of platelet concentrates has the potential to reduce the risk of bacterial contamination.

A general method for purification of deoxycytidine kinase.

Deoxycytidine kinase from a variety of animal tissues binds to Cibacron Blue 3G-A coupled to Sepharose CL-6B. The enzyme can be selectively eluted with ATP to yield single-step purifications of 17-89-fold. The affinity adsorbent is re-usable.