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At least 37 records · Page 2

Circulating Tumour Cell Numbers Correlate with Platelet Count and Circulating Lymphocyte Subsets in Men with Advanced Prostate Cancer: Data from the ExPeCT Clinical Trial (CTRIAL-IE 15-21)

Interactions between circulating tumour cells (CTCs) and platelets are thought to inhibit natural killer(NK)-cell-induced lysis. We attempted to correlate CTC numbers in men with advanced prostate cancer with platelet counts and circulating lymphocyte numbers. Sixty-one ExPeCT trial participants, divided into overweight/obese and normal weight groups on the basis of a BMI ≥ 25 or <25, were randomized to participate or not in a six-month exercise programme. Blood samples at randomization, and at three and six months, were subjected to ScreenCell filtration, circulating platelet counts were obtained, and flow cytometry was performed on a subset of samples (n = 29). CTC count positively correlated with absolute total lymphocyte count (r 2 = 0.1709, p = 0.0258) and NK-cell count (r 2 = 0.49, p < 0.0001). There was also a positive correlation between platelet count and CTC count (r 2 = 0.094, p = 0.0001). Correlation was also demonstrated within the overweight/obese group (n = 123, p < 0.0001), the non-exercise group (n = 79, p = 0.001) and blood draw samples lacking platelet cloaking (n = 128, p < 0.0001). By flow cytometry, blood samples from the exercise group (n = 15) had a higher proportion of CD3+ T-lymphocytes (p = 0.0003) and lower proportions of B-lymphocytes (p = 0.0264) and NK-cells (p = 0.015) than the non-exercise group (n = 14). These findings suggest that CTCs engage in complex interactions with the coagulation cascade and innate immune system during intravascular transit, and they present an attractive target for directed therapy at a vulnerable stage in metastasis.

60 APPLIED LIFE SCIENCES↗

Dietary calcium attenuates platelet aggregation and intracellular Ca2+ mobilization in spontaneously hypertensive rats

Spontaneously hypertensive rats (SHR) are known to be blood pressure sensitive to dietary calcium. The effects of dietary calcium on platelet aggregation and intracellular Ca2+ mobilization were assessed by turbidimetric methods and fura-2 methods, respectively, in washed platelets of SHR. Ca2+ ATPase activity was examined in aortic membrane fractions. Six weeks of dietary calcium supplementation attenuated the increase of systolic blood pressure (SBP 199 +/- 16 v 170 +/- 9 mm Hg, P < .001) and thrombin-induced platelet aggregation (84.5 +/- 3.7 v 73.7 +/- 7.4%, P < .004) at 9 weeks of age. The ionomycin-induced intracellular calcium ([Ca2+]i) peak in the absence of external Ca2+, which reflects [Ca2+]i storage size, and thrombin-evoked [Ca2+]i release from [Ca2+]i storage were decreased by 2.0% Ca diet (472 +/- 55 v 370 +/- 23 nmol/L, P < .001, 339 +/- 29 v 278 +/- 33 nmol/L, P < .002). In addition, SBP was positively correlated with platelet aggregation (r = 0.703, P = .0088), thrombin-evoked [Ca2+]i (r = 0.739, P = .0044), and ionomycin-induced [Ca2+]i (r = 0.591, P = .0415), respectively. However, there was no significant effect of dietary calcium on Ca2+-ATPase activity in aortic membranes. These results suggest that dietary calcium supplementation had a beneficial effect on platelets of SHR by attenuating [Ca2+]i mobilization from [Ca2+]i storage. The hypotensive effect of dietary calcium might be associated with attenuated [Ca2+]i mobilization in SHR.

NASA Discipline Regulatory Physiology↗

Hydrophilic Stent Coating Inhibits Platelet Adhesion on Stent Surfaces: Initial Results In Vitro

BackgroundEndovascular stents and flow diverter stents (FDS) have revolutionized the treatment of intradural aneurysms; however, the need for dual anti-platelet treatment (DAPT) limits their use and can cause additional issues. Therefore, there is a need to develop stent coatings that negate the need for DAPT.MethodsTwo different hydrophilic polymer coatings (HPC-I and HPC-II) were used to coat small nickel titanium plates to initially test the hydrophilic properties of these coatings when applied to nickel titanium. The plates were subsequently incubated with non-medicated whole blood from healthy volunteers for 10 min and stained with a CD61 immunofluorescent antibody that allows detection of adherent platelets. The coatings were applied to FDS wires and were again incubated with non-medicated whole blood from the same volunteers. Scanning electron microscopy was used to detect adherent platelets on the wire surface.ResultsThe HPC-II coating (1.12 ± 0.4%) showed a significantly lower CD61 +ve cell count (p ≤ 0.001) compared to both uncoated NiTi plates (48.61 ± 7.3%) and those with the HPC-I coating (mean 40.19 ± 8.9%). Minimal adherent platelets were seen on the FDS nickel titanium wires coated with the HPC-II compared to uncoated FDS under electron microscopy.ConclusionThere is a significant decrease in the number of adherent CD61 +ve platelets on nickel titanium surfaces coated with the HPC-II coating compared to uncoated surfaces. The coating can be successfully applied to the wires of flow diverters. The results of this study are promising with regard to the development of new anti-thrombogenic endovascular devices.

36 MATERIALS SCIENCE↗

Growth-Stimulating Effect of Human Platelets Stabilized with Silver Nanoparticles

We studied proliferative activity of multipotent mesenchymal stromal cells on collagen matrices containing platelets stabilized with silver nanoparticles. Dose-dependent stimulation of the growth of stromal cells without their structural damage was observed in the presence of stabilized platelets in proportion of 20-120 mln per 10{sup 5} multipotent mesenchymal stromal cells. The same doses of non-stabilized platelets produced less pronounced stimulation effect. In higher doses (≥180 mln) stabilized platelets inhibited cell proliferation and induced their damage. Stabilized platelets enhanced migration of multipotent mesenchymal stromal cells; after formation of the monolayer, they actively colonized deep layer of the collagen matrix. The formed monolayer of multipotent mesenchymal stromal cells survived over 14 days without appreciable cell damage.

60 APPLIED LIFE SCIENCES↗

Three-dimensional microstructural analysis of fragmentation behavior of platelet Al3Ti particles in Al-Al3Ti composite deformed by equal-channel angular pressing

Highlights: • Particle distribution in ECAPed Al-Al{sub 3}Ti composite depends on material flow of matrix. • Fragmentation of Al{sub 3}Ti particles by ECAP occurs along twin boundary in the particles. • Al{sub 3}Ti particles in the composite fragment after twin deformation of the particles. - Abstract: The fragmentation of platelet Al{sub 3}Ti particles in an Al-Al{sub 3}Ti composite by equal-channel angular pressing (ECAP) is investigated using three-dimensional (3D) microstructural evaluation and crystallography. The Al-Al{sub 3}Ti composite comprises coarse platelet Al{sub 3}Ti particles in an α-Al matrix. The ECAP of this composite is performed for up to 4 passes under processing routes A, B{sub c}, and C. Route B{sub c} produces the smallest Al{sub 3}Ti particle fragments in comparison with the fragments produced under routes A and C. This is because that route B{sub c} produces the largest number of shearing directions and shearing planes. Platelet Al{sub 3}Ti particle fragments in the specimen deformed by ECAP under route A are homogeneously distributed along the deformation axis. In contrast, in the specimens deformed by four ECAP passes under routes B{sub c} and C, the Al{sub 3}Ti particle fragments are gathered into several groups. The fragment groups have a length similar to the initial length of the platelet Al{sub 3}Ti particles. These spatial distributions of particle fragments can be explained by material flow of α-Al matrix during ECAP. Moreover, from 3D microstructural evaluation and crystallographic analysis of the Al{sub 3}Ti particle fragments, it is found that the Al{sub 3}Ti particles are preferentially fragmented on the twin boundary plane of {112}{sub Al3Ti}. Therefore, it is concluded that fragmentation of a platelet Al{sub 3}Ti particle by ECAP occurs along the twin boundary plane after twin deformation.

36 MATERIALS SCIENCE↗

Characterization of typical platelet injector flow configurations

A study to investigate the hydraulic atomization characteristics of several novel injector designs for use in liquid propellant rocket engines is presented. The injectors were manufactured from a series of thin stainless steel platelets through which orifices were very accurately formed by a photoetching process. These individual platelets were stacked together and the orifices aligned so as to produce flow passages of prescribed geometry. After alignment, the platelets were bonded into a single, 'platelet injector', unit by a diffusion bonding process. Because of the complex nature of the flow associated with platelet injectors, it was necessary to use experimental techniques, exclusively, throughout the study. Large scale models of the injectors were constructed from aluminum plates and the appropriate fluids were modeled using a glycerol-water solution. Stop-action photographs of test configurations, using spark-shadowgraph or stroboscopic back-lighting, are shown.

Hickox, C. E.↗

Development of the platelet micro-orifice injector

For some time to come, liquid rocket engines will continue to provide the primary means of propulsion for space transportation. The injector represents a key to the optimization of engine and system performance. The present investigation is concerned with a unique injector design and fabrication process which has demonstrated performance capabilities beyond that achieved with more conventional approaches. This process, which is called the 'platelet process', makes it feasible to fabricate injectors with a pattern an order of magnitude finer than that obtainable by drilling. The fine pattern leads to an achievement of high combustion efficiencies. Platelet injectors have been identified as one of the significant technology advances contributing to the feasibility of advanced dual-fuel booster engines. Platelet injectors are employed in the Space Shuttle Orbit Maneuvering System (OMS) engines. Attention is given to injector design theory as it relates to pattern fineness, a description of platelet injectors, and test data obtained with three different platelet injectors.

La Botz, R. J.↗

The stimulation of arachidonic acid metabolism in human platelets by hydrodynamic stresses

The effects of stimulating human platelets by thrombin and by hydrodynamic stresses on the platelets' arachidonic acid metabolism were investigated using (1-C-14)-arachidonic acid label and a specially designed viscometer that ensured laminar shear flow with a nearly uniform shear rate throughout the flow region. It was found that platelets activated by thrombin formed principally thromboxane A2, 12-hydroxy 5,8,10-heptadecatrienoic acid and 12-hydroxy 5,8,10,14-eicosatetraenoic acid (12-HETE). On the other hand, platelets activated by shear, formed only 12-HETE (although arachidonic acid metabolism was stimulated); no cyclooxygenase metabolites were detected. Results indicate that platelets may greatly increase their 12-HETE production when activated by passage through a high-stress region of the circulation, such as an atherosclerotic stenosis.

Rajagopalan, Sridhar↗

An autoanalyzer test for the quantitation of platelet-associated IgG

A new quantitative antiglobulin consumption (QAC) test for the measurement of platelet-associated IgG is described. In this test washed platelets are incubated with anti-IgG at a final dilution of 1:2 million. The unneutralized fraction of anti-IgG remaining in solution is then measured with an Autoanalyzer and soluble IgG is used for calibration. The dose-response curves depicting the percent neutralization of anti-IgG by platelets and by soluble IgG were compared in detail and found to be nearly identical, indicating that platelet-associated IgG can be accurately quantitated by this method. The mean IgG values were 2287 molecules/platelet for normal adults and 38,112 molecules/platelet for ITP patients. The Autoanalyzer QAC test is a sensitive and reproducible assay for the quantitation of platelet-associated IgG.

Levitan, Nathan↗

Formed platelets for low cost regeneratively cooled rocket combustion chambers

Formed platelet technology can be used to fabricate LO2/LH2-fueled rocket propulsion chambers with higher heat capacity, higher cycle life, and lower pressure drops, in conjunction with lower costs due to the application of high volume production methods. The formed-platelet combustor liner is an assembly of identical platelets, each of which is composed of diffusion-bonded, photoetched laminae with coolant flow passages; the platelets are also joined by diffusion bonding to form the combustor's circumference. Attention is given to the fabrication of a platelet combustion chamber generating 40 Klbf of thrust.

Burkhardt, W. M.↗

Incorporation of a circulating protein into megakaryocyte and platelet granules

To determine whether or not proteins circulating in plasma can be incorporated into megakaryocytes and platelets, horseradish peroxidase (HRP) was injected intravenously into guinea pigs and these cells were examined for its uptake by electron microscopy and cytochemistry. Enriched samples of megakaryocytes enabled ultrastructural analysis of large numbers of these rare cells. In megakaryocytes, 50% of alpha granules contained HRP between 75 min and 7 hr after injection. At 24 hr, 25% of the megakaryocyte granules were peroxidase-positive, less were positive by 48 hr, and there were none at 4 days. Thus, the findings demonstrate that a circulating protein can be endocytosed by megakaryocytes and rapidly packaged into alpha granules. Platelet granules also contain HRP by 7 hr after injection, and they can secrete it in response to thrombin. Unfortunately, our present studies do not allow us to distinguish between direct endocytosis by the platelet and/or shedding of new platelets from recently labeled megakaryocytes. It is concluded that while some alpha granule proteins are synthesized by megakaryocytes, others may be acquired from plasma by endocytosis. In addition to providing evidence that some of the proteins of alpha granules may be of exogenous origin, this study has allowed the definition of a pathway whereby plasma proteins may be temporarily sequestered in megakaryocytes before reentering the circulation in platelets.

Megakaryocytes/metabolism/ultrastructure↗

The effects of 7.5% NaCl/6% dextran 70 on coagulation and platelet aggregation in humans

The combination solution of 7.5% NaCl/6% dextran 70 (HSD) administered IV gives hemodynamic improvement in the treatment of hemorrhagic hypotension. Since earlier dextran solutions were reported to interfere with blood coagulation, the effects of HSD on the prothrombin time (PT), the activated partial thromboplastin time (APTT), platelet aggregation, and platelet concentration were studied. The HSD mixed with human plasma (1:5 and 1:10) slightly prolonged PT, but had no effect on the APTT, compared with saline controls. The HSD also decreased human platelet aggregation at the 1:5 dilution. In separate mixing studies, the hypertonic saline component of HSD was associated with the prolongation of PT and decreased platelet aggregation. The data from these studies indicate that at its proposed therapeutic dose, HSD is expected to have minimal effect on blood coagulation.

NASA Center ARC↗

RT-QuIC detection of chronic wasting disease prion in platelet samples of white-tailed deer

Chronic wasting disease (CWD) is a prion disease of captive and free-ranging cervids. Currently, a definitive diagnosis of CWD relies on immunohistochemistry detection of PrP Sc in the obex and retropharyngeal lymph node (RPLN) of the affected cervids. For high-throughput screening of CWD in wild cervids, RPLN samples are tested by ELISA followed by IHC confirmation of positive results. Recently, real-time quacking-induced conversion (RT-QuIC) has been used to detect CWD positivity in various types of samples. To develop a blood RT-QuIC assay suitable for CWD diagnosis, this study evaluated the assay sensitivity and specificity with and without ASR1-based preanalytical enrichment and NaI as the main ionic component in assay buffer. A total of 23 platelet samples derived from CWD-positive deer (ELISA +/IHC +) and 30 platelet samples from CWD-negative (ELISA-) deer were tested. The diagnostic sensitivity was 43.48% (NaCl), 65.22% (NaI), 60.87% (NaCl-ASR1) or 82.61% (NaI-ASR1). The diagnostic specificity was 96.67% (NaCl), 100% (NaI), 100% (NaCl-ASR1), or 96.67% (NaI-ASR1). The probability of detecting CWD prion in platelet samples derived from CWD-positive deer was 0.924 (95% CRI: 0.714, 0.989) under NaI-ASR1 experimental condition and 0.530 (95% CRI: 0.156, 0.890) under NaCl alone condition. The rate of amyloid formation (RFA) was greatest under the NaI-ASR1 condition at 10 -2 (0.01491, 95% CRI: 0.00675, 0.03384) and 10 -3 (0.00629, 95% CRI: 0.00283, 0.01410) sample dilution levels. Incorporation of ASR1-based preanalytical enrichment and NaI as the main ionic component signifcantly improved the sensitivity of CWD RT-QuIC on deer platelet samples. Blood test by the improved RT-QuIC assay may be used for antemortem and postmortem diagnosis of CWD.

59 BASIC BIOLOGICAL SCIENCES↗

N, N’-disubstitutedphenyl-4-ethoxyl benzene-1, 3-disulfonamides: design, synthesis, and evaluation of anti-platelet aggregation activity

According to the bio-isosterism theory, a series of N, N’-disubstitutedphenyl-4-ethoxylbenzene-1, 3-disulfonamides (5a-p) were designed and synthesized by two steps of reactions including chlorosulfonation and ammonolysis. The structures of all compounds have been confirmed by IR, {sup 1}H-NMR, {sup 13}C-NMR, and ESI-MS spectra. The in vitro anti-platelet aggregation activities were evaluated by Born’s test induced by adenosine diphosphate (ADP) and arachidonic acid (AA), respectively. The biological evaluation results revealed that compound 5h had the lowest IC{sub 50} value (0.32 μM) and the highest inhibition rate (40.9 %) that of three positive control agents clopidogrel (0.41 μM, 23.5 %), aspirin (0.53 μM, 28.9 %), and picotamide (0.76 μM, 32.7 %). Afterwards, compounds with higher activities were selected to further study in vitro cytotoxicity via cell counting kit-8 (CCK-8) assay. The cytotoxicity results indicated that compound 5h had simultaneously the lowest cytotoxicity, while other compounds had no significant relationship between the anti-platelet activities and cytotoxicities. Based on above in vitro anti-platelet activity data, the SAR (Structure Activity Relationship) of the target compounds was preliminarily summarized. In general, N, N’-disubstitutedphenyl-4-ethoxylbenzene-1, 3-disulfonamides have the potential of further study and very likely become safer and more effective anti-platelet agents.

60 APPLIED LIFE SCIENCES↗

Platelet cloaking of circulating tumour cells in patients with metastatic prostate cancer: Results from ExPeCT, a randomised controlled trial

Background: Circulating tumour cells (CTCs) represent a morphologically distinct subset of cancer cells, which aid the metastatic spread. The ExPeCT trial aimed to examine the effectiveness of a structured exercise programme in modulating levels of CTCs and platelet cloaking in patients with metastatic prostate cancer. Methods: Participants (n = 61) were randomised into either standard care (control) or exercise arms. Whole blood was collected for all participants at baseline (T0), three months (T3) and six months (T6), and analysed for the presence of CTCs, CTC clusters and platelet cloaking. CTC data was correlated with clinico-pathological information. Results: Changes in CTC number were observed within group over time, however no significant difference in CTC number was observed between groups over time. Platelet cloaking was identified in 29.5% of participants. A positive correlation between CTC number and white cell count (WCC) was observed (p = 0.0001), in addition to a positive relationship between CTC clusters and PSA levels (p = 0.0393). Conclusion: The presence of platelet cloaking has been observed in this patient population for the first time, in addition to a significant correlation between CTC number and WCC. Trial registration: ClincalTrials.gov identifier NCT02453139 .

60 APPLIED LIFE SCIENCES↗

Human blood platelets at microgravity

A set of freshly collected and separated human platelet suspensions were transported, in three types of plastic containers, on a 6 day, 2 hr mission of the orbiter Columbia to study the effect of prolonged exposure of human blood cells to microgravity. A controlled environment at a temperature of 22 + or - 1 deg with air flow was provided and another set of samples held on the ground acted as controls. Paired comparisons of platelets at ug versus controls at lxg revealed superior platelet survival at microgravity. When viewed in terms of plastic type, ug platelets in containers fabricated from PVC-TOTM displayed the best overall postflight viability.

Surgenor, D. MACN.↗

Sequential Dual Alignments Introduce Synergistic Effect on Hexagonal Boron Nitride Platelets for Superior Thermal Performance

Planarly aligning 2D platelets is challenging due to their additional orientational freedom compared to 1D materials. Here, this study reports a sequential dual-alignment approach, employing an extrusion-printing-induced shear force and rotating-magnetic-field-induced force couple for platelet planarly alignment in a yield-stress support bath. It is hypothesized that the partial alignment induced by a directional shear force facilitates subsequent axial rotation of the platelets for planar alignment under an external force couple, resulting in a synergistic alignment effect. This sequential dual-alignment approach achieves better planar alignment of 2D modified hexagonal boron nitride (mhBN). Specifically, the thermal conductivity of the 40 wt% mhBN/epoxy composite is significantly higher (692%) than that of unaligned composites, surpassing the cumulative effect of individual methods (only 133%) with a 5 times more synergistic effect. For 30, 40, and 50 wt% mhBN composites, the thermal conductivity values (5.9, 9.5, and 13.8 W m –1 K –1 ) show considerable improvement compared to the previously reported highest values (5.3, 6.6, and 8.6 W m –1 K –1 ). Additionally, a 3D mhBN/epoxy heat sink is printed and evaluated to demonstrate the feasibility of device fabrication. The approach enables the planar alignment of electrically or thermally conducting 2D fillers during 3D fabrication.

36 MATERIALS SCIENCE↗

Validation of process simulation workflow for thermosetting prepreg platelet molding compounds

Continuous carbon fiber prepreg slit and cut into rectangular platelets has proven to be a useful material for net shape molding of semi-structural and structural components in the aerospace and automotive industries. Furthermore, to assist the designer in use of these prepreg platelet molding compounds, sometimes called carbon fiber sheet molding compounds, simulation tools are required that can predict the as-manufactured fiber orientation state which has a significant impact on the resulting performance. Herein, an analysis workflow for design-enabling predictions is demonstrated for a double dome geometry with two different initial charge configurations. Here, the workflow is validated through comparison with experimental short shots, orientation state, and stiffness trends. Significantly, to complete the validation, a method is proposed for determining the confidence bounds on measured orientation state enhancing the results of optical microscopy which can only produce a small sample of platelet orientations.

36 MATERIALS SCIENCE↗