Platelet Shape Research Articles

Platelet ultrastructural changes stored at room temperature versus cold storage observed by electron microscopy and structured illumination microscopy

Our study seeks to provide a theoretical foundation for the clinical use of Cold-stored platelets (CSPs) by interpreting ultrastructural images and quantitatively analyzing structural changes. CSPs, room temperature-stored platelets (RTPs), and delayed cold stored platelets(Delayed-CSPs) were continuously observed using scanning electron microscopy and transmission electron microscopy at 8 time points. Super-resolution fluorescence microscopy was employed to observe changes in platelet microtubules and mitochondrial structure and function, while platelet counts, metabolism, and relevant functional indicators were measured concurrently. Quantitative statistical analysis of platelet size, morphology, canalicular systems, and five organelles was performed under electron microscopy. CSPs stored for 1 day, platelet shape changed from circular or elliptical to spherical, with size decreasing from 2.8 × 2.2µm to 2.0 × 2.0µm. CSPs occurred wrinkling and reorganization of platelet microtubule proteins, with organelles aggregating towards the central region. CSPs stored for 14 days and Delayed-CSPs for 10 days exhibited numerous structurally intact and active cells. The structure-intact active cells in both group were 92% . RTPs stored for 5 and 7 days showed minimal changes in size, a normal microtubule skeleton, and were primarily in a resting state. However, RTPs stored for 10 and 14 days displayed swelling, irregular disintegration of the microtubule skeleton, and the presence of membranous structures and vacuolated cells,the structure-intact active cells was only 45% and 7%, respectively. Our findings confirmed that the maximum storage time of platelets was 5-7 days for RTPs, within 10 days for Delayed-CSPs, and 14 days for CSPs.

Assay variables and early clinical evaluation of low-angle light scattering for platelet function analysis.

The recently developed platelet aggregation technique based on low-angle light scattering (LaSca) in diluted platelet-rich plasma (PRP) requires only a small sample volume and provides information about platelet aggregation and shape change. This study aimed to investigate the influence of preanalytical and analytical variables and to validate the method in a real-life pediatric hematology hospital setting. Platelet aggregation was induced by ADP in diluted PRP in the presence of 2mM calcium at 23°C. The study included healthy adults (n = 30), healthy children (n = 20), and pediatric patients with suspected or diagnosed platelet function abnormalities (n = 25). The assay parameters were stable for at least 3h after isolation of PRP and were sensitive to plasma dilution in the range of 2-8%. The initial aggregation velocity was significantly reduced in pediatric patients compared with healthy children (p < 0.05). ADP-induced light transmission amplitude was moderately correlated with LaSca amplitude of aggregation in healthy children (p = 0.52, p < 0.05) but not in pediatric patients. We standardized the protocol for platelet aggregation assessment by LaSca and characterized the influence of preanalytical and analytical variables on it.

LRRC8 complexes are adenosine nucleotide release channels regulating platelet activation and arterial thrombosis.

Platelet shape and volume changes are early mechanical events contributing to platelet activation and thrombosis. Here, we identify single-nucleotide polymorphisms in Leucine-Rich Repeat Containing 8 (LRRC8) protein subunits that form the Volume-Regulated Anion Channel (VRAC) which are independently associated with altered mean platelet volume. LRRC8A is required for functional VRAC in megakaryocytes (MKs) and regulates platelet volume, adhesion, and agonist-stimulated activation, aggregation, ATP secretion and calcium mobilization. MK-specific LRRC8A cKO mice have reduced arteriolar thrombus formation and prolonged arterial thrombosis without affecting bleeding times. Mechanistically, platelet LRRC8A mediates swell-induced ATP/ADP release to amplify agonist-stimulated calcium and PI3K-AKT signaling via P2X1, P2Y 1 and P2Y 12 receptors. Small-molecule LRRC8 channel inhibitors recapitulate defects observed in LRRC8A-null platelets in vitro and in vivo . These studies identify the mechanoresponsive LRRC8 channel complex as an ATP/ADP release channel in platelets which regulates platelet function and thrombosis, providing a proof-of-concept for a novel anti-thrombotic drug target.

Gelsolin controls the release of phosphatidylserine (PS)-positive microvesicles (MVs) from platelets

Upon activation by vascular injury or extracellular agonists, platelets undergo rapid change shape, a process regulated by the actin cytoskeleton and accessory proteins. Platelet shape change is accompanied by the secretion of hemostatic factors and immunomodulatory cytokines from their intracellular granules, as well as the release of microvesicles (MVs) containing pro-inflammatory cytokines and procoagulant phosphatidylserine (PS). However, the role of actin dynamics in MV generation remains unclear. In this study, we found that blocking actin polymerization with cytochalasin D attenuated the release of PS-positive MVs in human platelets stimulated by thrombin or the calcium ionophore A23187. The actin-severing protein gelsolin (Gsn) facilitates normal actin filament turnover in activated platelets. Platelets from Gsn-deficient (Gsn−/−) mice showed reduced MV release compared to platelets from control mice. These findings indicate that the proper dynamics of the actin cytoskeleton are essential for MV generation in platelets, which has implications for their pro-inflammatory and procoagulant functions.

Modification of Co3O4 by Al2O3: Influence on the reducibility

In this study, we used a coprecipitation method followed by calcination at 500 °C to synthesize undoped and Al3+-doped Co3O4 nanoparticles with different aluminum fractions (x = Al/(Co + Al) = 1/60, 1/30, 1/15, 1/75, 1/6 and 1/5). An addition of Al3+ ions led to a decrease in the average crystallite size from 29 to 11 nm, and growth of the specific surface area from 28 to 91 m2/g. TEM images indicated round and platelet shapes of the nanoparticles. According to HAADF-STEM combined with EDS elemental mapping, the platelet shape particles are Al3+-enriched, while the round shape particles are Al3+-depleted. The origin of Al3+ distribution over the oxide volume is conditioned by the state of the hydroxide precursor. It was shown by XRD that the coprecipitation yielded homogeneous hydroxides only for Al fractions x = 0 and x = 1/5. For the intermediate compositions, the precursors represent a mixture of Co6(CO3)2(OH)8*H2O and Co0.8Al0.2(OH)2(CO3)0.1*nH2O. On the TPR-H2 profiles, reduction peaks for three (Co,Al)3O4 oxides differing in the Al3+ concentration (y) can be found. Two of these oxides with y = 0 and y = 0.2 are formed from different hydroxides, and third one with y ∼0.05 is the result of their mutual interaction. In situ XRD allowed us to interpret the TPR peaks correctly and showed that the reduction of all the oxides occurs in two steps. In the first step, Co3+ → Co2+, and (Co1-yAly)3O4 oxides transform to (Co,Al)O. In the second step, Co2+ → Co0, and (Co,Al)O is reduced into metallic cobalt. In undoped Co3O4, Co3+ → Co2+ and Co2+ → Co0 reduction steps occur at T1 = 280 and T2 = 325 °C, respectively. For Al-depleted (Co1-yAly)3O4 (y ∼ 0.05 in the interior of particles), both reduction steps shift toward higher temperatures T1 = 305 and T2 = 405 °C, respectively. The reduction of Al-enriched (Co0.8Al0.2)3O4 is more difficult; first and second reduction steps occur at T1 = 345 and T2 = 610−690 °C. Therefore, Al3+ ions have a little effect on the first step and very significantly influence the second one. Additionally, it was shown by TEM that after the reduction at 700 °C metallic cobalt particles were surrounded by the Al-enriched oxide shell. Apparently, that is why the addition of even a small amount of Al3+ ions prevents a quick sintering of metallic cobalt observed for pure Co3O4.

Filamin A regulates platelet shape change and contractile force generation via phosphorylation of the myosin light chain.

Platelets are critical mediators of hemostasis and thrombosis. Platelets circulate as discs in their resting form but change shape rapidly upon activation by vascular damage and/or soluble agonists such as thrombin. Platelet shape change is driven by a dynamic remodeling of the actin cytoskeleton. Actin filaments interact with the protein myosin, which is phosphorylated on the myosin light chain (MLC) upon platelet activation. Actin-myosin interactions trigger contraction of the actin cytoskeleton, which drives platelet spreading and contractile force generation. Filamin A (FLNA) is an actin cross-linking protein that stabilizes the attachment between subcortical actin filaments and the cell membrane. In addition, FLNA binds multiple proteins and serves as a critical intracellular signaling scaffold. Here, we used platelets from mice with a megakaryocyte/platelet-specific deletion of FLNA to investigate the role of FLNA in regulating platelet shape change. Relative to controls, FLNA-null platelets exhibited defects in stress fiber formation, contractile force generation, and MLC phosphorylation in response to thrombin stimulation. Blockade of Rho kinase (ROCK) and protein kinase C (PKC) with the inhibitors Y27632 and bisindolylmaleimide (BIM), respectively, also attenuated MLC phosphorylation; our data further indicate that ROCK and PKC promote MLC phosphorylation through independent pathways. Notably, the activity of both ROCK and PKC was diminished in the FLNA-deficient platelets. We conclude that FLNA regulates thrombin-induced MLC phosphorylation and platelet contraction, in a ROCK- and PKC-dependent manner.

Effect of process parameter on the flowability of nanocrystalline CoNiCrAlY powder synthesized by mechanical milling

The present study concerns understanding the effect of process parameters on the characteristics and flowability of nanocrystalline CoNiCrAlY powder synthesized by mechanical milling. Mechanical milling has been conducted in a planetary ball mill with tungsten carbide (WC) ball, with ball to powder ratio of 10:1 at 300 rpm speed, using 1% stearic acid and toluene as process control agent (PCA) with time varying from 10 h to 36 h. The synthesized nanocrystalline powder were characterized by Scanning Electron Microscopy, X-ray Diffraction technique, X-ray Photoelectron Spectroscopy, and Differential Scanning Calorimetry. Subsequently, flowability in terms of Hausner ratio was assessed by Tap Density Tester. Average particle size of the powder was found to decrease from 33 μm to 22 μm after 10 h of milling and further increases to 43 μm and 38 μm after 25 and 36 h of milling, respectively, in stearic acid medium. However, in toluene medium particle size continuously decreases from 33 μm to 9.7 μm with increasing milling time. The particle morphology changes from spherical to platelet shape at low milling hours in both of the media. After 25 h of milling, the shape of the particles is nearly spherical for stearic acid and irregular for toluene used as a PCA. Crystallize size was found to decrease with increasing milling time from 147 nm to 7.7 nm and to 6.4 nm in stearic acid and toluene media, respectively. There was presence of γ, γʹ, β, hcp-Co, Al2O3 and AlOOH phases on the powder particles milled in both the medium. The measured Hausner ratio of the powders was found to vary from 1.18 to 1.32 in stearic acid medium, and was found to increase with increasing milling time. On the other hand, in toluene media flowability decreases (Hausner ratio increases from 1.33 to 1.44) with increasing milling time.

Abstract 119: LRRC8 Complex Is An ATP Release Channel Regulating Platelet Activation And Thrombosis

Platelet shape and volume changes are early mechanical events in platelet activation proposed to contribute to arterial thrombosis. Here, we identify single-nucleotide polymorphisms (SNPs) in four leucine-rich repeat containing protein subunits (LRRC8A, B, C, and D) expressed in human platelets that are independently associated with increased mean platelet volume in human genetic studies - implicating the volume-sensing LRRC8 channel complex as regulating platelet function in humans. We show LRRC8A is required for functional LRRC8/VRAC in megakaryocytes (MKs). MK-specific LRRC8A conditional knockout mice give rise to LRRC8A-null platelets with larger mean volumes. LRRC8A-null platelets and CRISPR-edited human LRRC8A KO MKs (hiPSC MKs) have reduced agonist -stimulated P-selectin exposure, and αIIbβ3 integrin activation. LRRC8A-null platelets exhibit impaired adhesion to collagen-coated surfaces, agonist-induced platelet aggregation and ATP secretion. MK-specific LRRC8A conditional knockout mice have reduced platelet thrombus formation in laser-induced cremaster arteriolar thrombosis and prolonged occlusion times in FeCl 3 -induced carotid arterial thrombosis, compared to control mice, without affecting tail bleeding times. Mechanistically, LRRC8A mediates swelling-induced ATP release to amplify agonist-stimulated calcium influx and aggregation in platelets. Treatment of platelets with small-molecule LRRC8 channel inhibitors recapitulate the defects in LRRC8A-null platelets. These studies identify the mechanoresponsive LRRC8 channel complex as a regulator of platelet function and thrombosis - providing a proof-of-concept for a novel anti-thrombotic drug target.

Impact of Shape Factor on a Couple Stress Flow of a Ternary Hybrid Nanofluid over an Inclined Flat Plate when Non-Uniform Heat Source and Thermal Radiation are Significant: A Darcy-Forchheimer Model

Abstract The study of fluid flow over an inclined flat plate finds applications in a diverse range of engineering fields including aerodynamics, energy production and automotive design. This study theoretically investigates the steady and radiative flow of a ternary hybrid nanofluid (Water + TiO2 + CoFe2O4 + MgO) with couple stress, using the Darcy-Forchheimer model. The flow occurs through a tilted flat plate and is subjected to irregular heat source parameter and entropy generation. The problem’s equations have been transformed into a collection of ordinary differential equations (ODEs), which has been skillfully resolved using the bvp4c solver. Graphs are utilized to elucidate outcomes for two instances of shape components, namely platelet and spherical. An escalation in the couple stress parameter (S) is demonstrated to be inversely related to the fluid velocity, resulting in a drop. Specifically, when 0.5 ≤ S ≤ 3, the friction factor exhibits a decline, with rates of 0.306201851 (for Platelet shape) and 0.304466755 (for Spherical shape). An intriguing observation reveals an augmentation in the generation of entropy as the volumetric fraction of TiO 2 rises. Upon investigation, it has been determined that when the Eckert number (Ecn) increases within the range of 0 ≤ Ecn ≤ 0.3, there is a significant reduction in the Nusselt number. Specifically, the decline is measured to be 0.328685192 for the platelet shape and 0.308939422 for the spherical shape. The utility of the Forchheimer number in regulating the fluid’s motion has been unveiled.

Ultrasoft platelet-like particles stop bleeding in rodent and porcine models of trauma.

Uncontrolled bleeding after trauma represents a substantial clinical problem. The current standard of care to treat bleeding after trauma is transfusion of blood products including platelets; however, donated platelets have a short shelf life, are in limited supply, and carry immunogenicity and contamination risks. Consequently, there is a critical need to develop hemostatic platelet alternatives. To this end, we developed synthetic platelet-like particles (PLPs), formulated by functionalizing highly deformable microgel particles composed of ultralow cross-linked poly (N-isopropylacrylamide) with fibrin-binding ligands. The fibrin-binding ligand was designed to target to wound sites, and the cross-linking of fibrin polymers was designed to enhance clot formation. The ultralow cross-linking of the microgels allows the particles to undergo large shape changes that mimic platelet shape change after activation; when coupled to fibrin-binding ligands, this shape change facilitates clot retraction, which in turn can enhance clot stability and contribute to healing. Given these features, we hypothesized that synthetic PLPs could enhance clotting in trauma models and promote healing after clotting. We first assessed PLP activity in vitro and found that PLPs selectively bound fibrin and enhanced clot formation. In murine and porcine models of traumatic injury, PLPs reduced bleeding and facilitated healing of injured tissue in both prophylactic and immediate treatment settings. We determined through biodistribution experiments that PLPs were renally cleared, possibly enabled by ultrasoft particle properties. The performance of synthetic PLPs in the preclinical studies shown here supports future translational investigation of these hemostatic therapeutics in a trauma setting.

Influence of thermal radiation and shape factor on a hybrid nanofluid flow over a permeable flat plate with cross-diffusion effects: An irreversibility analysis

Understanding and controlling the shape factors of nanoparticles in fluid flow problems is crucial for optimizing heat transfer, fluid dynamics, and various applications such as nanofluids, drug delivery systems, catalysis, and nanocomposites. By tailoring the shape of nanoparticles, researchers can manipulate their behavior, dispersion, and interaction with the fluid, thereby optimizing the desired outcomes in various fluid flow problems. The meticulous investigation uncovers the integration of entropy analysis in the investigation of the flow of convective magnetohydrodynamic hybrid nanofluid via a permeable flat surface, by considering the effects of Dufour, viscous dissipation, Soret, and thermal radiation. The governing equations are converted into a set of nonlinear ordinary differential equations using appropriate similarity transformations and then solved using the bvp4c solver, a MATLAB in-built function. We display the outcomes for three different shape factors: platelet, cylinder, and brick. The skin friction coefficient increases at a rate of 1.5089 for the brick shape, 1.5163 for the cylindrical shape, and 1.5212 for the platelet shape when the mixed convection parameter lies between 1 and 5. In this regard, it is important to point out that there is a direct connection between the increase in temperature transmission rate and a decline in the Dufour number ( Du ). It has been found that when 1 ≤ Du ≤ 2.4 , the Nusselt number decreases by 0.4786, 0.4618, and 0.4512 for brick, cylinder, and platelet shapes, respectively. A decline in the Sherwood number is noticed with an upsurge in the Soret number ( Sr ). There is a noticeable increase of 0.0442 in the Sherwood number for brick shape, 0.0438 for cylinder form, and 0.0435 for platelet form when 1 ≤ Du ≤ 2.4 . Furthermore, we have detected that an increase in the radiation parameter amplifies both the Bejan number and the profiles of entropy formation.

Platelet Activation Pathways Controlling Reversible Integrin αIIbβ3 Activation.

Background Agonist-induced platelet activation, with the integrin αIIbβ3 conformational change, is required for fibrinogen binding. This is considered reversible under specific conditions, allowing a second phase of platelet aggregation. The signaling pathways that differentiate between a permanent or transient activation state of platelets are poorly elucidated. Objective To explore platelet signaling mechanisms induced by the collagen receptor glycoprotein VI (GPVI) or by protease-activated receptors (PAR) for thrombin that regulate time-dependent αIIbβ3 activation. Methods Platelets were activated with collagen-related peptide (CRP, stimulating GPVI), thrombin receptor-activating peptides, or thrombin (stimulating PAR1 and/or 4). Integrin αIIbβ3 activation and P-selectin expression was assessed by two-color flow cytometry. Signaling pathway inhibitors were applied before or after agonist addition. Reversibility of platelet spreading was studied by microscopy. Results Platelet pretreatment with pharmacological inhibitors decreased GPVI- and PAR-induced integrin αIIbβ3 activation and P-selectin expression in the target order of protein kinase C (PKC) > glycogen synthase kinase 3 > β-arrestin > phosphatidylinositol-3-kinase. Posttreatment revealed secondary αIIbβ3 inactivation (not P-selectin expression), in the same order, but this reversibility was confined to CRP and PAR1 agonist. Combined inhibition of conventional and novel PKC isoforms was most effective for integrin closure. Pre- and posttreatment with ticagrelor, blocking the P2Y 12 adenosine diphosphate (ADP) receptor, enhanced αIIbβ3 inactivation. Spreading assays showed that PKC or P2Y 12 inhibition provoked a partial conversion from filopodia to a more discoid platelet shape. Conclusion PKC and autocrine ADP signaling contribute to persistent integrin αIIbβ3 activation in the order of PAR1/GPVI > PAR4 stimulation and hence to stabilized platelet aggregation. These findings are relevant for optimization of effective antiplatelet treatment.

Unsaturation and polar compounds of vegetable oils affect the properties of sunflower wax‐oleogels

AbstractAlthough it is important to understand the factors affecting the physical properties of oleogels for their application in food products, the effects of the kind of oils and components in them are not well understood. Previous studies on the effects of unsaturation and polar compounds of oils on the properties of oleogels reported inconsistent results, presumably, due to different gelling agents, experimental conditions, and different amounts of minor components in oils. In this study, a systematic study was conducted with 12 vegetable oils with and without polar compounds to understand these effects on the properties of 3% and 7% sunflower wax (SW)‐oleogels. In general, oils with higher unsaturation produced oleogels with lower gel strength. Polar compounds in oil negatively affected the gel strength. Melting and crystallization temperatures increased with increasing unsaturation of oil. The platelet shape of SW crystals was almost identical, regardless of the kind of oil. Solid wax content values were similar across the oleogel samples with 12 different oils. This study enlightens several factors affecting the physical properties of SW‐oleogels.Practical Application: This study provides information about the effects of unsaturation of oil and polar compounds in oil on firmness and melting properties of sunflower wax (SW)‐based oleogels. It was found that the firmness of SW‐oleogel can be significantly different (up to 65%) depending on vegetable oils used, and it decreased with increasing unsaturation of oil. It was also found that the firmness of the oleogel can be increased by up to 36% by removing polar compounds in oil. Solid wax content (SWC) of oleogels did show meaningful correlations with the firmness of SW‐oleogels. All the oleogels examined in this study had very similar shapes and sizes of SW crystals. For practical application, the unsaturation of oil and the polar compound content have to be considered when designing oleogels with desired textural properties.

In Situ Formation of Titanium Diboride/Magnesium Titanate Composites by Magnesiothermic-Based Combustion Synthesis

In situ formation of TiB2–Mg2TiO4 composites was investigated by combustion synthesis involving the solid-state reaction of Ti with boron and magnesiothermic reduction of B2O3. Certain amounts of MgO and TiO2 were added to the reactant mixtures of Ti/B/Mg/B2O3 to act as the moderator of highly exothermic combustion and a portion of the precursors to form Mg2TiO4. Two combustion systems were designed to ensure that synthesis reactions were sufficiently energetic to carry on self-sustainably, that is, in the mode of self-propagating high-temperature synthesis (SHS). Consistent with thermodynamic analyses, experimental results indicated that the increase in pre-added MgO and TiO2 decreased the combustion temperature and propagation velocity of the flame front. MgO was shown to have a stronger dilution effect on combustion exothermicity than TiO2, because the extent of magnesiothermic reduction of B2O3 was reduced in the MgO-added samples. In situ formation of the TiB2–Mg2TiO4 composite was achieved from both types of samples. It is believed that, in the course of the SHS progression, Mg2TiO4 was produced through a combination reaction between MgO and TiO2, both of which were entirely or partially generated from the metallothermic reduction of B2O3. The microstructure of the products exhibited fine TiB2 crystals in the shape of short rods and thin platelets that existed within the gaps of Mg2AlO4 grains. Both constituent phases were well distributed. A novel and efficient synthesis route, which is energy- and time-saving, for producing Mg2TiO4-containing composites was demonstrated.

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The primary goal of this investigation is to examine the heat and flow characteristics of a hybrid nanofluid consisting of silver (Ag) and aluminum oxide (Al2\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$_{2}$$\\end{document}O3)\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$_{3})$$\\end{document} nanoparticles over an unsteady radially stretching sheet embedded in porous medium. The investigation is conducted under the influence of several key parameters, namely joule heating, viscous dissipation, porous, slip, and suction. The technique of similarity transformations is used to transform the governing system of PDEs into nonlinear ODEs and the bvp4c solver is used to solve them numerically. The present study examines the influence of sphere and platelet shape nanoparticles on the temperature and velocity profiles. The outcomes are discussed through graphs and tables. A rise in the porous, slip, and suction parameters makes the velocity profile decrease gradually. The temperature escalates when Biot number, magnetic parameter, and Eckert number increase. As compared to sphere shapes, platelet-shaped nanoparticles exhibit the greatest heat transfer and flow. Results reveal that by using Ag-Al2\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$_{2}$$\\end{document}O3\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$_{3}$$\\end{document}/H2\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$_{2}$$\\end{document}O hybrid nanofluid with a volume fraction of 5%, the heat transfer enhancement of platelet shape nanoparticles increased by 11.88% than sphere-shaped nanoparticles. Overall, the platelet shape of nanoparticles offers distinctive advantages in various engineering applications, primarily due to their large surface area, anisotropic properties, and tunable surface chemistry. These properties make them versatile tools for improving the performance of materials and systems in engineering fields. The findings can contribute to the design and optimization of nanofluid-based systems in various engineering applications, such as heat exchangers, microfluidics, and energy conversion devices.

Role of the NO-GC/cGMP signaling pathway in platelet biomechanics

Cyclic guanosine monophosphate (cGMP) is a second messenger produced by the NO-sensitive guanylyl cyclase (NO-GC). The NO-GC/cGMP pathway in platelets has been extensively studied. However, its role in regulating the biomechanical properties of platelets has not yet been addressed and remains unknown. We therefore investigated the stiffness of living platelets after treatment with the NO-GC stimulator riociguat or the NO-GC activator cinaciguat using scanning ion conductance microscopy (SICM). Stimulation of human and murine platelets with cGMP-modulating drugs decreased cellular stiffness and downregulated P-selectin, a marker for platelet activation. We also quantified changes in platelet shape using deep learning-based platelet morphometry, finding that platelets become more circular upon treatment with cGMP-modulating drugs. To test for clinical applicability of NO-GC stimulators in the context of increased thrombogenicity risk, we investigated the effect of riociguat on platelets from human immunodeficiency virus (HIV)-positive patients taking abacavir sulfate (ABC)-containing regimens. Our results corroborate a functional role of the NO-GC/cGMP pathway in platelet biomechanics, indicating that biomechanical properties such as stiffness or shape could be used as novel biomarkers in clinical research.

Antiplatelet and Antithrombotic Activities of Lespedeza cuneata via Pharmacological Inhibition of Integrin αIIbβ3, MAPK, and PI3K/AKT Pathways and FeCl3-Induced Murine Thrombosis.

Cardiovascular diseases (CVDs) have been the major cause of mortality all around the globe. Lespedeza cuneata abbreviated as L. cuneata with the authority name of Dumont de Courset (G. Don) is a perennial flowering plant commonly grown in Asian countries such as Korea, Japan, China, and Taiwan. We aimed to investigate the L. cuneata extract's antiplatelet and antithrombotic properties as GC-MS analysis indicated that the extract contained short-chain fatty acids, which have been reported to possess beneficial cardiovascular effects. L. cuneata was extracted using water, 50% EtOH, 70% EtOH, and 100% EtOH. For in vitro antiplatelet analysis, washed platelets were prepared and incubated with L. cuneata with 200 μg/mL of 50% EtOH in the presence of 1 mM of CaCl2 for 1 minute followed by agonist (collagen 2.5 μg/mL or ADP 10 μM or thrombin 0.1 U/mL) stimulation for 5 minutes over light transmission aggregometer. Scanning electron microscopy was performed to assess platelet shape change. ATP release and intracellular calcium mobilization were quantified to assess the granular content. Fibrinogen-binding assay and clot retraction assay assessed integrin αIIbβ3-mediated inside-out and outside-in signaling. Protein phosphorylation expression was investigated by western blot analysis. Finally, the in vivo antithrombotic efficacy was investigated by oral dosage of L. cuneata 200 and 400 mg/kg and aspirin 100 mg/kg for 7 days, and tail bleeding and FeCl3-induced murine thrombus model were performed. In vitro platelet aggregation and platelet shape change were dose-dependently suppressed by L. cuneata. Calcium mobilization, dense granules secretion, integrin αIIbβ3-mediated inside-out and outside-in signaling, and protein phosphorylation of MAPK and PI3K/Akt pathways were significantly inhibited. In vivo assays revealed that L. cuneata prevents side effects of synthetic drugs via nonsignificantly increasing bleeding time and improving coronary artery blood flow and animal survival. Our results demonstrate that L. cuneata exhibited potent antiplatelet and antithrombotic effects and can be considered a potential herbal medicine with cardioprotective effects.

Efficiency assessment of thermal radiation utilizing flow of advanced nanocomposites on riga plate

Utilizing nanostructured materials to optimize thermal processes for renewable energy is an important strategy that has been applied in numerous engineering disciplines for diverse interventions such as photovoltaic thermal systems. Ternary nanofluids have demonstrated potential for improving the cooling capabilities of this kind of system. The current research focuses on enhancing the thermal performance of convective flow of a water-based trihybrid nanofluid across a Riga plate incorporating carbon nanotubes in spherical, graphene in cylindrical and Al2O3 in platelet shapes. Several mechanisms like non-linear heat production, thermal radiation, velocity and thermal slip conditions have all been investigated regarding the flow describing equations. Due to the high complexity of the problem modeling and numerical methods, conventional techniques to the parametric work that incorporate various model factors can fail to derive accurate solutions. This study assesses the ability of soft computing techniques to determine the behavior of a problem with various connected parameters to resolve this issue. With help from numerical scheme Lobatto IIIa technique reference data, the artificial neural network is trained. Furthermore, a constructive technique using artificial neural networks (Levenberg–Marquardt) is used to quantitatively determine the rate of heat transmission on the surface of Riga plates. With the use of correlation, fitting analysis, regression, mean squared error, absolute error, and error histogram, the prediction performance of the proposed algorithms is examined. The Cfx and Nux reach convergence with mean squared errors close to 10−8 and 10−10 after 626 and 100 epochs. The investigation illustrates the benefits of using soft computing methods to accurately investigate the behavior of complex flow models. The most important discovery is that, in comparison to mono- and binary-hybrid nanofluids, ternary hybrid nanofluids have a greater thermal responsiveness.

Investigation of platelet activation and calcium store release by a topical hemostatic xerogel dressing for improved blood clotting

AbstractAdequate topical hemostatic dressings are required as first‐line approach for reduction of uncontrolled hemorrhage, a leading cause of mortalities. Platelets play a major role during blood clotting to prevent hemorrhage during injuries. Here, we demonstrate the mechanisms activated through protease activating receptor 1 (PAR1), a platelet membrane protein in response to a porous composite xerogel dressing incorporating SiNPs (size 122 ± 10 nm) and calcium (2.5 mM), characterized by scanning electron microscopy–energy dispersive x‐ray spectroscopy and Fourier transform infrared spectroscopy. The composite displayed 13.9‐fold improved blood clotting index in comparison to commercial dressing. The composite displayed increased platelet aggregation due to development of well‐formed pseudopodia as compared to bare xerogel, SFLLRN (a thrombin mimic), adenosine di‐phosphate (a platelet activator), and heparin (a thrombin inhibitor). Further, PAR1 gene was significantly upregulated in model A549 epithelial cell line (1.2‐fold) and human platelets (1.4‐fold). The composite enhanced calcium release and its extrusion in A549 cells. Upregulation of PAR1 on the platelet surface and calcium release are crucial for platelet shape change and aggregation. The data indicate that xerogel composite containing SiNPs and calcium enhanced blood clotting through activation of PAR1. Such dressings can provide a potential hemostatic solution to reduce blood loss, disability, and mortality during surgery and trauma care.

Cold temperature induces a TRPM8-independent calcium release from the endoplasmic reticulum in human platelets.

The detection of temperature by the human sensory system is life-preserving and highly evolutionarily conserved. Platelets are sensitive to temperature changes and are activated by a decrease in temperature, akin to sensory neurons. However, the molecular mechanism of this temperature-sensing ability is unknown. Yet, platelet activation by temperature could contribute to numerous clinical sequelae, most importantly to reduced quality of ex vivo-stored platelets for transfusion. In this multidisciplinary study, we present evidence for the expression of the temperature-sensitive ion channel transient receptor potential cation channel subfamily member 8 (TRPM8) in human platelets and precursor cells. We found the TRPM8 mRNA and protein in MEG-01 cells and platelets. Inhibition of TRPM8 prevented temperature-induced platelet activation and shape change. However, chemical agonists of TRPM8 did not seem to have an acute effect on platelets. When exposing platelets to below-normal body temperature, we detected a cytosolic calcium increase which was independent of TRPM8 but was completely dependent on the calcium release from the endoplasmic reticulum. Because of the high interindividual variability of TRPM8 expression, a population-based approach should be the focus of future studies. Our study suggests that the cold response of platelets is complex and TRPM8 appears to play a role in early temperature-induced activation of platelets, while other mechanisms likely contribute to later stages of temperature-mediated platelet response.