Platelet dysfunction reversal with cold-stored vs room temperature-stored platelet transfusions.

ABSTRACT: Platelets are stored at room temperature for 5 to 7 days (room temperature-stored platelets [RSPs]). Because of frequent and severe shortages, the US Food and Drug Administration recently approved up to 14-day cold-stored platelets (CSPs) in plasma. However, the posttransfusion function of CSPs is unknown and it is unclear which donors are best suited to provide either RSPs or CSPs. In this study, we sought to evaluate the posttransfusion platelet function and its predictors for platelets stored for the maximum approved storage times (7-day RSPs and 14-day CSPs) in healthy volunteers on acetylsalicylic acid (ASA). We conducted a randomized crossover study in 10 healthy humans. Individuals donated 1 platelet unit, stored at either 22°C or 4°C based on randomization. Before transfusion, participants ingested ASA to inhibit endogenous platelets. Transfusion recipients were tested for platelet function and lipid mediators. Platelet units were tested for lipid mediators only. A second round of transfusion with the alternative product was followed by an identical testing sequence. RSPs reversed platelet inhibition significantly better in αIIbß3 integrin activation-dependent assays. In contrast, CSPs in recipients led to significantly more thrombin generation, which was independent of platelet microparticles. Lysophosphatidylcholine-O species levels predicted the procoagulant capacity of CSPs. In contrast, polyunsaturated fatty acid concentrations predicted the aggregation response of RSPs. In summary, we provide, to our knowledge, the first efficacy data of extended-stored CSPs in plasma. Our results suggest that identifying ideal RSP and CSP donors is possible, and pave the way for larger studies in the future. This trial is registered at www.ClinicalTrials.gov as #NCT0511102.

Low-Dose Dietary Fish Oil Improves RBC Deformability without Improving Post-Transfusion Recovery in Mice.

Long-chain polyunsaturated fatty acids (LC-PUFAs) are important modulators of red blood cell (RBC) rheology. Dietary LC-PUFAs are readily incorporated into the RBC membrane, improving RBC deformability, fluidity, and hydration. Female C57BL/6J mice consumed diets containing increasing amounts of fish oil (FO) ad libitum for 8 weeks. RBC deformability, filterability, and post-transfusion recovery (PTR) were evaluated before and after cold storage. Lipidomics and lipid peroxidation markers were evaluated in fresh and stored RBCs. High-dose dietary FO (50%, 100%) was associated with a reduction in RBC quality (i.e., in vivo lifespan, deformability, lipid peroxidation) along with a reduced 24 h PTR after cold storage. Low-dose dietary FO (6.25-12.5%) improved the filterability of fresh RBCs and reduced the lipid peroxidation of cold-stored RBCs. Although low doses of FO improved RBC deformability and reduced oxidative stress, no improvement was observed for the PTR of stored RBCs. The improvement in RBC deformability observed with low-dose FO supplementation could potentially benefit endurance athletes and patients with conditions resulting from reduced perfusion, such as peripheral vascular disease.

Loss of 12-Lipoxygenase Improves the Post-Transfusion Function of Stored Platelets.

BACKGROUND: Platelets for transfusion are stored for 5 to 7 days. Previous studies have shown that HETE levels in the storage bag negatively correlate with platelet performance in vivo, suggesting that the dysregulation of bioactive lipid mediators may contribute to the storage lesion. In the current study, we sought to understand how genetic deletion and pharmacological inhibition of 12-LOX (12-lipoxygenase) affects platelets during storage and after transfusion. METHODS: Platelets from 12-LOX+/+ (wild-type [WT]) and 12-LOX-/- mice were stored for 24 and 48 hours and profiled using liquid chromatography-tandem mass spectrometry-multiple reaction monitoring or transfused into thrombocytopenic hIL4R (human interleukin 4 receptor)-transgenic mice. Platelet function was assessed by flow cytometry and in vivo thrombosis and hemostasis models. To test the role of the COX-1 (cyclooxygenase-1) pathway, donor mice were treated with acetylsalicylic acid. Human platelets were treated with the 12-LOX inhibitor, VLX-1005, or vehicle, stored, and transfused to NOD/SCID (nonobese diabetic/severe combined immunodeficiency) mice. RESULTS: Polyunsaturated fatty acids increased significantly in stored platelets from 12-LOX-/- mice, whereas oxylipin concentrations were significantly higher in WT platelets. After transfusion to thrombocytopenic mice, we observed significantly more baseline αIIbß3 integrin activation in 12-LOX-/- platelets than in WT platelets. Stored platelets from 12-LOX-/- mice occluded vessels significantly faster than stored WT platelets. In hemostasis models, significantly more stored 12-LOX-/- than WT platelets accumulated at the site of venous injury leading to reduced blood loss. Inhibition of COX-1 abrogated both increased integrin activation and thromboxane generation in stored 12-LOX-/- platelets, highlighting the critical role of this pathway for improved post-transfusion function. Consistent with our mouse studies, human platelets stored with VLX-1005, showed increased integrin activation compared with vehicle-treated platelets after transfusion. CONCLUSIONS: Deleting 12-LOX improves the post-transfusion function of stored murine platelets by increasing thromboxane generation through COX-1-dependent arachidonic acid metabolism. Future studies should determine the feasibility and safety of 12-LOX-inhibited platelets transfused to humans.

Recovery via SfM photogrammetry of latent footprint impressions in carpet.

Impression evidence retained in carpet is usually recovered, if at all, in two dimensions via a vertical photograph. Here, we show that recovery is also possible via SfM photogrammetry and this gives good quality results that allow digital measurements both in the x-y plane and by depth (z-axis). This study focuses on recovery from polypropylene carpets which are widespread due to their resistance to wear and low cost. We show how traces can be recovered using both SfM photogrammetry and conventional photography with illumination provided via a crime scene light source. Experiments show that traces are retained for considerable time periods if left undisturbed, in excess of four weeks, but are quickly lost in under 8 hours by subsequent footfall. A simple simulation shows how the movement of an individual can be determined from carpet traces and the value of 3D recovery is illustrated via a set of experiments conducted with barefoot traces. We draw attention to the fact that 3D models allow a more statistical-based approach to be taken to match bare footprints at crime scenes. SfM photogrammetry is shown to provide a useful compliment to existing techniques and therefore worthy of further experimentation and potentially operational use.

Recovery of 3D footwear impressions using a range of different techniques.

Three-dimensional (plastic) footwear impressions are frequently found at, or in the vicinity of a crime scene, and may provide a valuable form of evidence or intelligence. This paper compares the traditional methods of casting and/or two-dimensional photography with Structure from Motion (SfM) photogrammetry. We focus both on the recovery of class characteristics (sole pattern) and randomly acquired characteristics caused by damage. We examine how different recovery techniques influence visualization of outsole features and discuss what effect this may have on evidential value. Five shoes and their associated three-dimensional impressions made in both sand and soil were compared using a grid system and tread descriptors commonly used in the UK. We conclude that within the limitations of this study SfM photogrammetry allows superior levels of visualization of both class and randomly acquired characteristics, giving a better definition in detail in some instances. The use of SfM as a complementary approach can therefore lead to a potential increase in evidential value.

Empirical Evaluation of the Reliability of Photogrammetry Software in the Recovery of Three-Dimensional Footwear Impressions.

This paper examines the reliability of Structure from Motion (SfM) photogrammetry as a tool in the capture of forensic footwear marks. This is applicable to photogrammetry freeware DigTrace but is equally relevant to other SfM solutions. SfM simply requires a digital camera, a scale bar, and a selection of oblique photographs of the trace in question taken at the scene. The output is a digital three-dimensional point cloud of the surface and any plastic trace thereon. The first section of this paper examines the reliability of photogrammetry to capture the same data when repeatedly used on one impression, while the second part assesses the impact of varying cameras. Using cloud to cloud comparisons that measure the distance between two-point clouds, we assess the variability between models. The results highlight how little variability is evident and therefore speak to the accuracy and consistency of such techniques in the capture of three-dimensional traces. Using this method, 3D footwear impressions can, in many substrates, be collected with a repeatability of 97% with any variation between models less than ~0.5 mm.