What can we learn from senescent platelets, their transcriptomes and proteomes?

Research into the natural aging process of platelets has garnered much research interest in recent years, and there have long been associations drawn between the proportion of newly formed platelets in the circulation and the risk of thrombosis. However, these observations have largely been demonstrated in patient groups in which there may be underlying systemic changes that effect platelet function. Recent advances in technology have allowed in-depth analysis of differently aged platelets isolated from the peripheral blood of healthy individuals and have demonstrated that aged platelets, often referred to as senescent platelets, undergo extensive changes in the transcriptome and proteome. Ultimately, these changes result in platelets whose functions have deteriorated such that they cannot partake in hemostatic responses to the same extent as newly formed platelets. Here, we review transcriptomic and proteomic research in platelet aging in the context of health and how this research sheds light upon alterations in platelet structure and function

Platelet ageing: A review.

Platelet ageing is an area of research which has gained much interest in recent years. Newly formed platelets, often referred to as reticulated platelets, young platelets or immature platelets, are defined as RNA-enriched and have long been thought to be hyper-reactive. This latter view is largely rooted in associations and observations in patient groups with shortened platelet half-lives who often present with increased proportions of newly formed platelets. Evidence from such groups suggests that an increased proportion of newly formed platelets is associated with an increased risk of thrombotic events and a reduced effectiveness of standard anti-platelet therapies. Whilst research has highlighted the existence of platelet subpopulations based on function, size and age within patient groups, the common intrinsic changes which occur as platelets age within the circulation are only just being explored. By understanding the changes that occur during the natural ageing processes of platelets, we may be able to identify the triggers for alterations in platelet life span and platelet reactivity. Here we review research on platelet ageing in the context of health and disease, paying particular attention to the experimental approaches taken and the robustness of conclusions that can be drawn

Clustering of tau-immunoreactive pathology in chronic traumatic encephalopathy

Chronic traumatic encephalopathy (CTE) is a neurodegenerative disorder which may result from repetitive brain injury. A variety of tau-immunoreactive pathologies are present, including neurofibrillary tangles (NFT), neuropil threads (NT), dot-like grains (DLG), astrocytic tangles (AT), and occasional neuritic plaques (NP). In tauopathies, cellular inclusions in the cortex are clustered within specific laminae, the clusters being regularly distributed parallel to the pia mater. To determine whether a similar spatial pattern is present in CTE, clustering of the tau-immunoreactive pathology was studied in the cortex, hippocampus, and dentate gyrus in 11 cases of CTE and 7 cases of Alzheimer’s disease neuropathologic change (ADNC) without CTE. In CTE: (1) all aspects of tau-immunoreactive pathology were clustered and the clusters were frequently regularly distributed parallel to the tissue boundary, (2) clustering was similar in two CTE cases with minimal co-pathology compared with cases with associated ADNC or TDP-43 proteinopathy, (3) in a proportion of cortical gyri, estimated cluster size was similar to that of cell columns of the cortico-cortical pathways, and (4) clusters of the tau-immunoreactive pathology were infrequently spatially correlated with blood vessels. The NFT and NP in ADNC without CTE were less frequently randomly or uniformly distributed and more frequently in defined clusters than in CTE. Hence, the spatial pattern of the tau-immunoreactive pathology observed in CTE is typical of the tauopathies but with some distinct differences compared to ADNC alone. The spread of pathogenic tau along anatomical pathways could be a factor in the pathogenesis of the disease

Recovery From a Forward Falling Slip: Measurement of Dynamic Stability and Strength Requirements Using a Split-Belt Instrumented Treadmill

Aim: Falls commonly occur from trips and slips while walking. Recovery strategies from trips and backward falling slips have been extensively studied. However, until recently, forward falling slips (FFSs) have been considered less dangerous and have been understudied. This study aimed first to create an application to realistically simulate FFSs using a split-belt instrumented treadmill and then to understand the biomechanical requirements for young adults to recover from an FFS. Methods: We developed a semi-automatic custom-made application on D-Flow that triggered FFSs by briefly and unexpectedly increasing the speed (a = 5 m·s−2) of the right belt during stance. To validate the protocol, we tested against criteria defined for an ecologically and experimentally valid FFS: unexpected occurrence of the slip, increased foot velocity, forward loss of balance during the slip and consistent perturbation timing. We evaluated the recovery strategies of 17 young adults by measuring dynamic stability, joint moments and ground reaction force (GRF) vector angles before, during and on 15 steps following the FFS. Results: The application successfully triggered FFSs, according to the criteria we defined. Participants' balance returned to normal for a minimum of three consecutive steps in 10.9 (7.0) steps. Recovery from the FFSs was characterised by larger hip flexor and knee extensor moments to support the centre of mass during the slip, and a longer first recovery step with large hip extensor moments to arrest the fall followed by large knee extensor moments to raise and advance the centre of mass into the next step (p < 0.001 compared with normal gait). Subsequent steps progressively returned to normal. Conclusion: This is the first study to experimentally simulate FFSs meeting the aforementioned criteria, and to measure their effects on the dynamic balance and kinetic parameters. The split-belt instrumented treadmill proved a promising tool to better study the mechanisms of falls and recovery. The required large hip and knee joint moments generally agree with findings on trips and backward falling slips and provide an indication of the functional capacities that should be targeted in fall-prevention interventions. These findings should be used to better understand and target the mechanisms of balance loss and falls in older adults following FFSs

Immature platelet dynamics are associated with clinical outcomes after major trauma.

BACKGROUND: Major trauma results in dramatic changes in platelet behavior. Newly-formed platelets are more reactive than older platelets, but their contributions to hemostasis and thrombosis after severe injury have not been previously evaluated. OBJECTIVES: To determine the relationship between immature platelet metrics and plasma thrombopoietin with clinical outcomes after major injury. METHODS: Prospective observational cohort study of adult trauma patients. Platelet counts and the immature platelet fraction (IPF) were measured at admission, 24 hours, 72 hours and 7 days post-injury. Thromboelastometry was performed at admission. Plasma thrombopoietin, c-Mpl and GPIbα were quantified in a separate cohort. The primary outcome was in-hospital mortality; secondary outcomes were venous thromboembolic events (VTE) and multiple organ dysfunction (MODS). RESULTS: On admission, immature platelet counts (IPC) were significantly lower in non-survivors (n=40) compared to survivors (n=236; 7.3x109/L vs 10.6x109/L, p=0.009), but IPF did not differ. Similarly, impaired platelet function on thromboelastometry was associated with lower admission IPC (9.1x109/L vs 11.9x109/L, p<0.001). However, at later timepoints we observed significantly higher IPF and IPC in patients who developed VTE (21.0x109/L vs 11.1x109/L, p=0.02) and prolonged MODS (20.9 x109/L vs 11x109/L, p=0.003) compared to those who did not develop complications. Plasma thrombopoietin levels at admission were significantly lower in in non-survivors (p<0.001), in patients with MODS (p<0.001) and in those who developed VTE (p=0.04). CONCLUSIONS: Lower levels of immature platelets in the acute phase after major injury are associated with increased mortality, whereas higher immature platelet levels at later timepoints may predispose to thrombosis and multiple organ dysfunction

Thrombosis Is Reduced by Inhibition of COX-1, but Unaffected by Inhibition of COX-2, in an Acute Model of Platelet Activation in the Mouse

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Combination of cyclic nucleotide modulators with P2Y12 receptor antagonists as anti-platelet therapy

"This is the peer reviewed version of the following article: Armstrong, PC, Ferreira, PM, Chan, MV, et al. Combination of cyclic nucleotide modulators with P2Y12 receptor antagonists as anti‐platelet therapy. J Thromb Haemost. 2020 https://doi.org/10.1111/jth.14826 which has been published in final form at   https://doi.org/10.1111/jth.14826BACKGROUND: Endothelium-derived prostacyclin and nitric oxide elevate platelet cyclic nucleotide levels and maintain quiescence. We previously demonstrated that a synergistic relationship exists between cyclic nucleotides and P2Y12 receptor inhibition. A number of clinically approved drug classes can modulate cyclic nucleotide tone in platelets including activators of NO-sensitive guanylyl cyclase (GC) and phosphodiesterase (PDE) inhibitors. However, the doses required to inhibit platelets produce numerous side effects including headache. OBJECTIVE: We investigated using GC-activators in combination with P2Y12 receptor antagonists as a way to selectively amplify the anti-thrombotic effect of both drugs. METHODS: In vitro light transmission aggregation and platelet adhesion under flow were performed on washed platelets and platelet rich plasma. Aggregation in whole blood and a ferric chloride-induced arterial thrombosis model were also performed. RESULTS: The GC-activator BAY-70 potentiated the action of the P2Y12 receptor inhibitor prasugrel active metabolite in aggregation and adhesion studies and was associated with raised intra-platelet cyclic nucleotide levels. Furthermore, mice administered sub-maximal doses of the GC activator cinaciguat together with the PDE inhibitor dipyridamole and prasugrel, showed significant inhibition of ex vivo platelet aggregation and significantly reduced in vivo arterial thrombosis in response to injury without alteration in basal carotid artery blood flow. CONCLUSIONS: Using in vitro, ex vivo, and in vivo functional studies, we show that low dose GC activators synergize with P2Y12 inhibition to produce powerful anti-platelet effects without altering blood flow. Therefore, modulation of intra-platelet cyclic nucleotide levels alongside P2Y12 inhibition can provide a strong, focused anti-thrombotic regimen while minimizing vasodilator side effects

Recovery From a Forward Falling Slip: Measurement of Dynamic Stability and Strength Requirements Using a Split-Belt Instrumented Treadmill

Data Availability Statement: The datasets generated for this study are available on request to the corresponding author.Copyright © 2020 Debelle, Harkness-Armstrong, Hadwin, Maganaris and O'Brien. Aim: Falls commonly occur from trips and slips while walking. Recovery strategies from trips and backward falling slips have been extensively studied. However, until recently, forward falling slips (FFSs) have been considered less dangerous and have been understudied. This study aimed first to create an application to realistically simulate FFSs using a split-belt instrumented treadmill and then to understand the biomechanical requirements for young adults to recover from an FFS. Methods: We developed a semi-automatic custom-made application on D-Flow that triggered FFSs by briefly and unexpectedly increasing the speed (a = 5 m·s−2) of the right belt during stance. To validate the protocol, we tested against criteria defined for an ecologically and experimentally valid FFS: unexpected occurrence of the slip, increased foot velocity, forward loss of balance during the slip and consistent perturbation timing. We evaluated the recovery strategies of 17 young adults by measuring dynamic stability, joint moments and ground reaction force (GRF) vector angles before, during and on 15 steps following the FFS. Results: The application successfully triggered FFSs, according to the criteria we defined. Participants' balance returned to normal for a minimum of three consecutive steps in 10.9 (7.0) steps. Recovery from the FFSs was characterised by larger hip flexor and knee extensor moments to support the centre of mass during the slip, and a longer first recovery step with large hip extensor moments to arrest the fall followed by large knee extensor moments to raise and advance the centre of mass into the next step (p < 0.001 compared with normal gait). Subsequent steps progressively returned to normal. Conclusion: This is the first study to experimentally simulate FFSs meeting the aforementioned criteria, and to measure their effects on the dynamic balance and kinetic parameters. The split-belt instrumented treadmill proved a promising tool to better study the mechanisms of falls and recovery. The required large hip and knee joint moments generally agree with findings on trips and backward falling slips and provide an indication of the functional capacities that should be targeted in fall-prevention interventions. These findings should be used to better understand and target the mechanisms of balance loss and falls in older adults following FFSs.Minerva Research Labs Ltd. (London, UK); Liverpool John Moores University