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Sample records for platelet concentrates feasibility

  1. Ultraviolet irradiation of platelet concentrates: Feasibility in transfusion practice

    SciTech Connect

    Andreu, G.; Boccaccio, C.; Lecrubier, C.; Fretault, J.; Coursaget, J.; LeGuen, J.P.; Oleggini, M.; Fournel, J.J.; Samama, M. )

    1990-06-01

    Ultraviolet (UV)-B irradiation abolishes lymphocyte functions (the ability to respond and to stimulate) in mixed lymphocyte culture (MLC). This effect may have practical application in the prevention or reduction of transfusion-induced alloimmunization against HLA class I antigens. To study this, platelet concentrates (PCs) were obtained with a cell separator, suspended in autologous plasma in a final volume of 400 mL, and transferred into a large (22 X 30 cm) cell culture bag. This plastic showed a good transmittance of UV-B rays at 310 nm (54%). PCs were placed between two quartz plates (surface of irradiation = 25 X 37 cm), and the two sides were irradiated simultaneously. Energy delivered to the surface of the plastic bag was automatically monitored. The ability to respond (in MLC and to phytohemagglutinin) and to stimulate allogeneic lymphocytes was completely abolished with energy of 0.75 J per cm2 (irradiation time less than 3 min). The temperature increase during irradiation was negligible. Platelet aggregation (collagen, adrenalin, ADP, arachidonic acid, ristocetin) was not impaired if UV-B energy was below 3 J per cm2. Recovery and survival of autologous 111In-labeled platelets were studied in four volunteers; no differences were found between UV-B-treated (1.5 J/cm2) platelets and untreated platelets. These results show that a large-scale clinical trial using UV-B-irradiated PCs to prevent HLA alloimmunization is feasible.

  2. Comparison of platelet counting technologies in equine platelet concentrates.

    PubMed

    O'Shea, Caitlin M; Werre, Stephen R; Dahlgren, Linda A

    2015-04-01

    (1) To compare the performance of 4 platelet counting technologies in equine platelet concentrates and (2) to evaluate the ability of the Magellan platelet rich plasma (PRP) system to concentrate equine platelets. Experimental study to assess method agreement. Adult mixed breed horses (n = 32). Acid citrate dextrose-A anti-coagulated whole blood was collected and PRP produced using the Magellan system according to the manufacturer's instructions. Platelets were quantified using 4 counting methods: optical scatter (Advia 2120), impedance (CellDyn 3700), hand counting, and fluorescent antibody flow cytometry. Platelet concentrations were compared using Passing and Bablok regression analyses and mixed model ANOVA. Significance was set at P < .05. Platelet concentrations measured in identical PRP samples were consistently higher for the Advia 2120 than the CellDyn 3700. Systematic and proportional biases were observed between these 2 automated methods when analyzed by regression analysis of the larger sample size. No bias (systematic or proportional) was observed among any of the other counting methods. Despite the bias detected between the 2 automated systems, there were no significant differences on average among the 4 counting methods evaluated, based on the ANOVA. The Magellan system consistently generated high platelet concentrations as well as higher than expected WBC concentrations. The Magellan system delivered desirably high platelet concentrations; however, WBC concentrations may be unacceptably high for some orthopedic applications. All 4 platelet counting methods tested were equivalent on average and therefore suitable for quantifying platelets in equine PRP used for clinical applications. © Copyright 2014 by The American College of Veterinary Surgeons.

  3. Pooled platelet concentrates: an alternative to single donor apheresis platelets?

    PubMed

    Pietersz, R N I

    2009-10-01

    Three types of platelet concentrates (PC) are compared: PC either processed with the platelet-rich plasma (PRP) or the Buffy coat (BC) method from whole blood units and PC obtained by apheresis. Leuko-reduction (LR) pre-storage is advocated to improve quality of the PC during storage and reduce adverse reactions in recipients. Standardization of methods allow preparation of PC with comparable yields of approximately 400 x 10(9) platelets in pooled non-LR-PRP, approximately 370 x 10(9) in pooled LR-BC-PC and in LR apheresis PC the number of platelets can be targeted on 350 x 10(9) or more with devices of various manufacturers. While viral transmission can be prevented by outstanding laboratory tests, the risk of bacterial contamination should be reduced by improved arm disinfection, deviation of the first 20-30 ml of blood and culture or rapid detection assays of the PC pre-issue. In a large prospective multicenter trial no significant difference was observed between cultures of apheresis PC (n = 15,198): 0.09% confirmed positive units versus 0.06% in pooled BC-PC (n = 37,045), respectively. Though platelet activation as measured by CD62 expression may differ in vitro in PC obtained with various apheresis equipment, and also between PC processed with the two whole blood methods there is scarce literature about the clinical impact of these findings. In conclusion the final products of LR-PC derived from whole blood or obtained by apheresis can be comparable, provided the critical steps of the processing method are identified and covered and the process is in control.

  4. Platelet concentration in platelet concentrates and periodontal regeneration-unscrambling the ambiguity

    PubMed Central

    Suchetha, A.; Lakshmi, P.; Bhat, Divya; Mundinamane, Darshan B.; Soorya, K. V.; Bharwani, G. Ashit

    2015-01-01

    Context: Platelet-rich-plasma (PRP) and Platelet-rich-fibrin (PRF) are extensively used autologous platelet concentrates in periodontal regeneration, and PRF has a better efficacy as compared to PRP. The rationale for this difference has often been attributed to the difference in the structure of the fibrin matrix. However, the effect of concentration of platelets on the regenerative potential of these concentrates is obscure. Aims: The study was conducted to evaluate and compare, clinically and radiographically, the efficacy of PRF and PRP in the treatment of periodontal endosseous defects and to assess the effect of platelet concentration on periodontal regeneration. Materials and Methods: Twenty intrabony defects were selected and divided into two groups randomly by the coin toss method. Group I received PRP and Group II subjects were treated with PRF. The platelet counts in PRP and PRF were analyzed. Clinical and radiological parameters were assessed at baseline and 3, 6, and 9 months postoperatively. Statistical Analysis: Kruskal–Wallis Chi-square test, Wilcoxon signed rank test, t-test, and Spearman's rank correlation were used for statistical analysis of data. Results: There was statistically significant improvement in all the parameters in the two groups except in relation to gingival recession. There was a statistically significant difference between the platelet count in Group I and Group II (P = 0.002). Conclusion: PRP and PRF appear to have nearly comparable effects in terms of periodontal regeneration. The concentration of platelets appears to play a paradoxical role in regeneration. The regenerative potential of platelets appears to be optimal within a limited range. PMID:26681857

  5. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part II: platelet-related biologic features.

    PubMed

    Dohan, David M; Choukroun, Joseph; Diss, Antoine; Dohan, Steve L; Dohan, Anthony J J; Mouhyi, Jaafar; Gogly, Bruno

    2006-03-01

    Platelet-rich fibrin (PRF) belongs to a new generation of platelet concentrates, with simplified processing and without biochemical blood handling. In this second article, we investigate the platelet-associated features of this biomaterial. During PRF processing by centrifugation, platelets are activated and their massive degranulation implies a very significant cytokine release. Concentrated platelet-rich plasma platelet cytokines have already been quantified in many technologic configurations. To carry out a comparative study, we therefore undertook to quantify PDGF-BB, TGFbeta-1, and IGF-I within PPP (platelet-poor plasma) supernatant and PRF clot exudate serum. These initial analyses revealed that slow fibrin polymerization during PRF processing leads to the intrinsic incorporation of platelet cytokines and glycanic chains in the fibrin meshes. This result would imply that PRF, unlike the other platelet concentrates, would be able to progressively release cytokines during fibrin matrix remodeling; such a mechanism might explain the clinically observed healing properties of PRF.

  6. Formed platelet combustor liner construction feasibility, phase A

    NASA Astrophysics Data System (ADS)

    Hayes, W. A.; Janke, D. E.

    1992-09-01

    Environments generated in high pressure liquid rocket engines impose severe requirements on regeneratively cooled combustor liners. Liners fabricated for use in high chamber pressures using conventional processes suffer from limitations that can impair operational cycle life and can adversely affect wall compatibility. Chamber liners fabricated using formed platelet technology provide an alternative to conventional regeneratively cooled liners (an alternative that has many attractive benefits). A formed platelet liner is made from a stacked assembly of platelets with channel features. The assembly is diffusion bonded into a flat panel and then three-dimensionally formed into a section of a chamber. Platelet technology permits the liner to have very precisely controlled and thin hot gas walls and therefore increased heat transfer efficiency. Further cooling efficiencies can be obtained through enhanced design flexibility. These advantages translate into increased cycle life and enhanced wall compatibility. The increased heat transfer efficiency can alternately be used to increase engine performance or turbopump life as a result of pressure drop reductions within the regeneratively cooled liner. Other benefits can be obtained by varying the materials of construction within the platelet liner to enhance material compatibility with operating environment or with adjoining components. Manufacturing cost savings are an additional benefit of a formed platelet liner. This is because of reduced touch labor and reduced schedule when compared to conventional methods of manufacture. The formed platelet technology is not only compatible with current state-of-the art combustion chamber structural support and manifolding schemes, it is also an enabling technology that allows the use of other high performance and potentially low cost methods of construction for the entire combustion chamber assembly. The contract under which this report is submitted contains three phases: (1) phase

  7. Formed platelet combustor liner construction feasibility, phase A

    NASA Technical Reports Server (NTRS)

    Hayes, W. A.; Janke, D. E.

    1992-01-01

    Environments generated in high pressure liquid rocket engines impose severe requirements on regeneratively cooled combustor liners. Liners fabricated for use in high chamber pressures using conventional processes suffer from limitations that can impair operational cycle life and can adversely affect wall compatibility. Chamber liners fabricated using formed platelet technology provide an alternative to conventional regeneratively cooled liners (an alternative that has many attractive benefits). A formed platelet liner is made from a stacked assembly of platelets with channel features. The assembly is diffusion bonded into a flat panel and then three-dimensionally formed into a section of a chamber. Platelet technology permits the liner to have very precisely controlled and thin hot gas walls and therefore increased heat transfer efficiency. Further cooling efficiencies can be obtained through enhanced design flexibility. These advantages translate into increased cycle life and enhanced wall compatibility. The increased heat transfer efficiency can alternately be used to increase engine performance or turbopump life as a result of pressure drop reductions within the regeneratively cooled liner. Other benefits can be obtained by varying the materials of construction within the platelet liner to enhance material compatibility with operating environment or with adjoining components. Manufacturing cost savings are an additional benefit of a formed platelet liner. This is because of reduced touch labor and reduced schedule when compared to conventional methods of manufacture. The formed platelet technology is not only compatible with current state-of-the art combustion chamber structural support and manifolding schemes, it is also an enabling technology that allows the use of other high performance and potentially low cost methods of construction for the entire combustion chamber assembly. The contract under which this report is submitted contains three phases: (1) phase

  8. Optimized preparation method of platelet-concentrated plasma and noncoagulating platelet-derived factor concentrates: maximization of platelet concentration and removal of fibrinogen.

    PubMed

    Araki, Jun; Jona, Masahiro; Eto, Hitomi; Aoi, Noriyuki; Kato, Harunosuke; Suga, Hirotaka; Doi, Kentaro; Yatomi, Yutaka; Yoshimura, Kotaro

    2012-03-01

    Platelet-rich plasma (PRP) has been clinically used as an easily prepared growth factor cocktail that can promote wound healing, angiogenesis, and tissue remodeling. However, the therapeutic effects of PRP are still controversial, due partly to the lack of optimized and standardized preparation protocols. We used whole blood (WB) samples to optimize the preparation protocols for PRP, white blood cell-containing (W-PRP), platelet-concentrated plasma (PCP), and noncoagulating platelet-derived factor concentrate (PFC). PRP and W-PRP were most efficiently collected by 10 min centrifugation in a 15-mL conical tube at 230-270 g and 70 g, respectively. To prepare PCP, platelets were precipitated by centrifugation of PRP at >2300 g, 90% of supernatant plasma was removed, and the platelets were resuspended. For preparation of noncoagulating PFC, the supernatant was replaced with one-tenth volume of saline, followed by platelet activation with thrombin. Platelet (before activation) and platelet-derived growth factor (PDGF)-BB (after activation) concentrations in PCP were approximately 20 times greater than those in WB, whereas PFC contained a 20-times greater concentration of platelets before platelet activation and a 50-times greater concentration of PDGF-BB without formation of a fibrin gel after platelet activation than WB. Surprisingly, total PDGF-BB content in the PFC was twice that of activated WB, which suggested that a substantial portion of the PDGF-BB became trapped in the fibrin glue, and replacement of plasma with saline is crucial for maximization of platelet-derived factors. As an anticoagulant, ethylene di-amine tetra-acetic acid disodium inhibited platelet aggregation more efficiently than acid citrate dextrose solution, resulting in higher nonaggregated platelet yield and final PDGF-BB content. These results increase our understanding of how to optimize and standardize preparation of platelet-derived factors at maximum concentrations.

  9. Optimized Preparation Method of Platelet-Concentrated Plasma and Noncoagulating Platelet-Derived Factor Concentrates: Maximization of Platelet Concentration and Removal of Fibrinogen

    PubMed Central

    Araki, Jun; Jona, Masahiro; Eto, Hitomi; Aoi, Noriyuki; Kato, Harunosuke; Suga, Hirotaka; Doi, Kentaro; Yatomi, Yutaka

    2012-01-01

    Abstract Platelet-rich plasma (PRP) has been clinically used as an easily prepared growth factor cocktail that can promote wound healing, angiogenesis, and tissue remodeling. However, the therapeutic effects of PRP are still controversial, due partly to the lack of optimized and standardized preparation protocols. We used whole blood (WB) samples to optimize the preparation protocols for PRP, white blood cell-containing (W-PRP), platelet-concentrated plasma (PCP), and noncoagulating platelet-derived factor concentrate (PFC). PRP and W-PRP were most efficiently collected by 10 min centrifugation in a 15-mL conical tube at 230–270 g and 70 g, respectively. To prepare PCP, platelets were precipitated by centrifugation of PRP at >2300 g, 90% of supernatant plasma was removed, and the platelets were resuspended. For preparation of noncoagulating PFC, the supernatant was replaced with one-tenth volume of saline, followed by platelet activation with thrombin. Platelet (before activation) and platelet-derived growth factor (PDGF)-BB (after activation) concentrations in PCP were approximately 20 times greater than those in WB, whereas PFC contained a 20-times greater concentration of platelets before platelet activation and a 50-times greater concentration of PDGF-BB without formation of a fibrin gel after platelet activation than WB. Surprisingly, total PDGF-BB content in the PFC was twice that of activated WB, which suggested that a substantial portion of the PDGF-BB became trapped in the fibrin glue, and replacement of plasma with saline is crucial for maximization of platelet-derived factors. As an anticoagulant, ethylene di-amine tetra-acetic acid disodium inhibited platelet aggregation more efficiently than acid citrate dextrose solution, resulting in higher nonaggregated platelet yield and final PDGF-BB content. These results increase our understanding of how to optimize and standardize preparation of platelet-derived factors at maximum concentrations. PMID

  10. EXTENDED STORAGE OF PLATELET-RICH PLASMA PREPARED PLATELET CONCENTRATES IN PLASMA OR PLASMALYTE

    PubMed Central

    Slichter, Sherrill J.; Bolgiano, Doug; Corson, Jill; Jones, Mary Kay; Christoffel, Todd

    2010-01-01

    Background Using bacterial detection or pathogen reduction, extended platelet storage may be licensed if platelet viability is maintained. FDA's post-storage platelet acceptance guidelines are that autologous stored platelet recoveries and survivals should be ≥66% and ≥58%, respectively, of each donor's fresh platelet data. Study Design And Methods Non-leukoreduced platelet concentrates were prepared from whole blood donations. Autologous platelet concentrates from 62 subjects were stored in 100% plasma (n=44) or 20% plasma/80% Plasmalyte (n=18), an acetate based, non-glucose containing crystalloid solution previously used for platelet storage.(1-3) Fresh platelets were obtained on the day the donor's stored platelets were to be transfused. The fresh and stored platelets were alternately radiolabeled with either 51Chromium or 111Indium, and in vitro measurements were performed on the stored platelets. Results FDA's platelet recovery criterion was met for 7 days of plasma storage, but platelet survivals maintained viability for only 6 days. Plasmalyte stored platelets did not meet either acceptance criteria after 6 days of storage. After 7 days of storage, platelet recoveries averaged 43 ± 4% and 30 ± 4% and survivals 4.1 ± 0.4 days and 2.0 ± 0.2 days for plasma and Plasmalyte-stored platelets, respectively (p=0.03 for recoveries and p<0.001 for survivals). Post-storage platelet recoveries correlated with the commonly-used in vitro platelet quality measurements of HSR and Annexin V binding, while survivals correlated with ESC, morphology score, and pH. Conclusion There is a progressive decrease in recoveries and survivals of plasma stored platelets over time. Platelet viability is better maintained in plasma than Plasmalyte. PMID:20456703

  11. Platelet activation of platelet concentrates derived from buffy coat and apheresis methods.

    PubMed

    Ali, Soleimany Ferizhandy

    2011-02-01

    Preparation for storage may cause platelet activation. The quality of platelet concentrates plays an important role in transfusion therapy. Platelet concentrates are produced by different centrifugation methods; buffy coat (buffy coat-derived platelet concentrates-BC) and plateletpheresis (apheresis-derived platelet concentrates-APC). Their quality was assessed using the following parameters: platelet, WBC and RBC counts pH, volume, platelet factor 4 (PF4) and Annexin V. The present paper compares the quality of both platelet preparations in vitro. In this experimental study, 30 platelet concentrates were harvested with the Haemonetics MCS plus and 30 units via the buffy coat (BC) method. The percentages of Annexin V expression, PF4 levels, platelet, WBC and RBC counts, pH and volume were measure immediately after collection and after 3 days of storage. During storage for up to 3 days, BC units displayed, no significant pH or RBC, difference in comparison with apheresis preparations (p>0.05). During storage for up to 3 days, BC units displayed a significant increase in the PF4 and Annexin V expression, compared to the apheresis preparations on day three (p<0.05). The kinetics of PF4 and Annexin V levels are influenced by the method used to prepare platelets for storage. The different levels of PF4 and Annexin V in BCs and APCs clearly demonstrates a progressive activation of BC platelets exceeding that of APC. However, in vivo studies should be performed to confirm these findings.

  12. Use of random versus apheresis platelet concentrates.

    PubMed

    Andreu, G; Vasse, J; Sandid, I; Tardivel, R; Semana, G

    2007-12-01

    The respective use of random (RPC) and apheresis (APC) platelet concentrates is highly heterogeneous among countries, ranging from 10 to 98% RPC in countries supposed to provide a similar transfusion service to patients. Moreover, when considering each country in the past 10 years, one can observe that some have changed their policy, switching from a majority of APC to RPC or vice versa. This presentation intends to analyse which factors may impact such decisions. For many years, the only available platelet component was a RPC obtained from whole blood donation by a two centrifugation steps process, the "platelet rich plasma" or PRP method. Since the beginning of the 1970s, APCs became available, with in fact many different techniques leading to many APCs that may not be equivalent. Since the end of the 1980s, a new method of RPC preparation was developed, using the buffy-coat (BC-PC), providing a blood component with highly preserved platelet functions as compared to RPCs prepared by the PRP technique. Finally, the use of each of these components either native, or leuco-reduced, or suspended in a storage solution, or processed with a pathogen inactivation technique adds new layers of complexity to compare them. Innumerable references can be found in the literature describing in vitro functional parameters of platelet concentrates. Although it is clear that BC-RPC retain much more their in vitro functions than PRP-RPC, indicating that no one should use the latter any more, it is much more difficult to distinguish differences between other PCs. Conversely, only a very few studies have been published related to a comparison of clinical efficacy of RPC versus APC, the endpoints being mainly CCI. Similarly to the in vitro studies, although RPC prepared with the PRP method show the lowest CCIs, no clear difference exists between "modern" RPC and APC. Another factor that may impact policy decision is the occurrence of adverse reactions in recipients. When considering

  13. Comparison of point-of-care methods for preparation of platelet concentrate (platelet-rich plasma).

    PubMed

    Weibrich, Gernot; Kleis, Wilfried K G; Streckbein, Philipp; Moergel, Maximilian; Hitzler, Walter E; Hafner, Gerd

    2012-01-01

    This study analyzed the concentrations of platelets and growth factors in platelet-rich plasma (PRP), which are likely to depend on the method used for its production. The cellular composition and growth factor content of platelet concentrates (platelet-rich plasma) produced by six different procedures were quantitatively analyzed and compared. Platelet and leukocyte counts were determined on an automatic cell counter, and analysis of growth factors was performed using enzyme-linked immunosorbent assay. The principal differences between the analyzed PRP production methods (blood bank method of intermittent flow centrifuge system/platelet apheresis and by the five point-of-care methods) and the resulting platelet concentrates were evaluated with regard to resulting platelet, leukocyte, and growth factor levels. The platelet counts in both whole blood and PRP were generally higher in women than in men; no differences were observed with regard to age. Statistical analysis of platelet-derived growth factor AB (PDGF-AB) and transforming growth factor β1 (TGF-β1) showed no differences with regard to age or gender. Platelet counts and TGF-β1 concentration correlated closely, as did platelet counts and PDGF-AB levels. There were only rare correlations between leukocyte counts and PDGF-AB levels, but comparison of leukocyte counts and PDGF-AB levels demonstrated certain parallel tendencies. TGF-β1 levels derive in substantial part from platelets and emphasize the role of leukocytes, in addition to that of platelets, as a source of growth factors in PRP. All methods of producing PRP showed high variability in platelet counts and growth factor levels. The highest growth factor levels were found in the PRP prepared using the Platelet Concentrate Collection System manufactured by Biomet 3i.

  14. A flow cytometric method for platelet counting in platelet concentrates.

    PubMed

    van der Meer, Pieter F; Karssing-van Leeuwen, Willy; Kurtz, Jim; Spengler, Hans-Peter; Blair, AbbeJane; Devine, Dana; Harrison, Paul; Lambrecht, Bernd; VandenBroeke, Tania; de Wildt, Janny; de Korte, Dirk

    2012-01-01

    The platelets (PLTs) in PLT concentrates are counted with hematology analyzers, but varying results among different hematology analyzers are observed, making comparisons very difficult. Due to the absence of red blood cells in PLT concentrates, the International Council for Standardization in Hematology (ICSH) reference method was modified to be used for PLT concentrates and validated in an international comparative study. Five PLT samples were shipped to eight participating centers of the Biomedical Excellence for Safer Transfusion (BEST) Collaborative and counted on the same day. PLTs were stained with fluorescein isothiocyanate-labeled anti-CD41a in tubes (TruCount, BD Biosciences), measured on a flow cytometer, and analyzed with a uniform template. These samples were also counted on 15 hematology analyzers. The ICSH method and newly developed BEST method yielded PLT counting results with less than 1% difference (not significant). The intercenter coefficient of variation (CV) of the BEST method was on average 6.3% versus 7.6% on average for hematology analyzers. The CV of individual hematology analyzers was on average 0.9%, which was considerably lower than for the flow cytometers with a mean of 3.7%. The BEST flow cytometric method has a smaller intercenter CV and a smaller center-to-center deviation from the group mean compared to hematology analyzers. Conversely, individual hematology analyzers are more precise than the flow cytometric method. Thus, the flow cytometric method provides a calibration tool to allow comparisons between centers, but there is no need to replace routine counting with hematology analyzers. © 2011 American Association of Blood Banks.

  15. Platelet concentrates, from whole blood or collected by apheresis?

    PubMed

    van der Meer, Pieter F

    2013-04-01

    Platelet concentrates can be isolated from donated whole blood with the platelet-rich plasma-method or the buffy coat-method. Alternatively, platelets can be obtained by apheresis, harvesting the platelets but returning all other cells to the donor. The quality and characteristics of platelets during storage are affected by a number of factors, such as anticoagulant, centrifugation and processing after collection, and pre- or post storage pooling, but when comparing literature on the various methods, most differences balance out. To have sufficient platelets to treat an adult patient, whole-blood-derived platelet concentrates need pooling of multiple donations, thereby increasing the risk of infectious agent transmission at least two-fold as compared with apheresis units. Allo immunization rates, acute reaction rates, and transfusion related acute lung injury rates are not different. Apheresis donation procedures have fewer adverse events. All these factors need to be considered and weighed when selecting a method of platelet collection for a blood center.

  16. Platelet counting in platelet concentrates with various automated hematology analyzers.

    PubMed

    Dijkstra-Tiekstra, Margriet J; Kuipers, Willeke; Setroikromo, Airies C; de Wildt-Eggen, Janny

    2007-09-01

    Hematology analyzers use impedance, optical, and/or immunologic techniques for counting platelets (PLTs). PLT counting in whole blood has been validated thoroughly; however, this is not the case for PLT counting in PLT concentrates (PCs), in which red cells (RBCs) are absent. Therefore, this study is focused on PLT counting in PCs to study use of ethylenediaminetetraacetate (EDTA), carryover, and accuracy of the analyzers. In total six hematology analyzers (AcT 8, Beckman Coulter; ADVIA 2,120, Bayer; Cell-Dyn 4,000, Abbott; Onyx, Beckman Coulter; K4,500, Sysmex; and XT 2,000i, Sysmex) were tested for PLT counting. PC samples with various PLT concentrations were made (0-1,700 x 10(9)/L) and measured 10 times. Carryover was determined five times. PC samples (1,000 x 10(9) PLTs/L) in EDTA tubes showed significantly higher PLT counts than samples in "dry" tubes for all analyzers except for the Cell-Dyn 4,000 with the impedance technique. Carryover was not more than 0.3 percent for all analyzers. The K4,500 showed the most accurate results, whereas the Cell-Dyn 4,000 with the impedance technique had low accuracy due to an overestimation of more than 20 percent. Most tested analyzers seemed to be suitable for counting PLTs in PCs. All hematology analyzers should be validated for counting PLTs in absence of RBCs as is the case in PCs, in addition to validation of PLT counting in whole blood.

  17. EXTENDED STORAGE OF BUFFY-COAT PLATELET CONCENTRATES IN PLASMA OR A PLATELET ADDITIVE SOLUTION

    PubMed Central

    Slichter, Sherrill J.; Bolgiano, Doug; Corson, Jill; Jones, Mary Kay; Christoffel, Todd; Bailey, S. Lawrence; Pellham, Esther

    2014-01-01

    Background Platelet concentrates prepared from whole blood in the U.S. are made using the platelet-rich-plasma (PRP) method. The platelet concentrates must be made within 8 hours of blood collection and stored for only 5 days. In Europe and Canada, platelet concentrates are made using the buffy-coat (BC) method from whole blood held overnight at 22°C and storage times may be up to 7 days. Our studies were designed to determine how long BC platelets can be stored in plasma or Plasmalyte while meeting the FDA’s post-storage viability criteria. Study Design, Materials, And Methods Normal subjects donated whole blood that was stored at 22°C for 22 ± 2 hours prior to preparation of BC platelets. Platelets were stored for 5 to 8 days in either plasma or Plasmalyte concentrations of 65% or 80%. Radiolabeled autologous stored versus fresh platelet recoveries and survivals were assessed as well as post-storage in vitro assays. Results BC platelets stored in either plasma or 65% Plasmalyte met FDA post-storage platelet recovery criteria for 7 days but survivals for only 6 days, while storage in 80% Plasmalyte gave very poor results. Both stored platelet recoveries and survivals correlated with the same donor’s fresh results, but the correlation was much stronger between recoveries than survivals. In vitro measures of extent of shape change, morphology score, and pH best predicted post-storage platelet recoveries, while annexin V binding best predicted platelet survivals. Conclusion BC platelets stored in either plasma or 65% Plasmalyte meet FDA’s post-storage viability criteria for 6 days. PMID:24673482

  18. Characterization of Platelet Concentrates Using Dynamic Light Scattering

    PubMed Central

    Labrie, Audrey; Marshall, Andrea; Bedi, Harjot; Maurer-Spurej, Elisabeth

    2013-01-01

    Summary Background Each year, millions of platelet transfusions save the lives of cancer patients and patients with bleeding complications. However, between 10 and 30% of all platelet transfusions are clinically ineffective as measured by corrected count increments, but no test is currently used to identify and avoid these transfusions. ThromboLUX® is the first platelet test intended to routinely characterize platelet concentrates prior to transfusion. Methods ThromboLUX is a non-invasive, optical test utilizing dynamic light scattering to characterize a platelet sample by the relative quantity of platelets, microparticles, and other particles present in the sample. ThromboLUX also determines the response of platelets to temperature changes. From this information the ThromboLUX score is calculated. Increasing scores indicate increasing numbers of discoid platelets and fewer microparticles. ThromboLUX uses calibrated polystyrene beads as a quality control standard, and accurately measures the size of the beads at multiple temperatures. Results Results from apheresis concentrates showed that ThromboLUX can determine the microparticle content in unmodified samples of platelet concentrates which correlates well with the enumeration by flow cytometry. ThromboLUX detection of microparticles and microaggregates was confirmed by microscopy. Conclusion ThromboLUX provides a comprehensive and novel analysis of platelet samples and has potential as a noninvasive routine test to characterize platelet products to identify and prevent ineffective transfusions. PMID:23652319

  19. Cryopreservation of buffy-coat-derived platelet concentrates in dimethyl sulfoxide and platelet additive solution.

    PubMed

    Johnson, L N; Winter, K M; Reid, S; Hartkopf-Theis, T; Marks, D C

    2011-04-01

    Platelets prepared in plasma can be frozen in 6% dimethyl sulfoxide (Me(2)SO) and stored for extended periods at -80°C. The aim of this study was to reduce the plasma present in the cryopreserved product, by substituting plasma with platelet additive solution (PAS; SSP+), whilst maintaining in vitro platelet quality. Buffy coat-derived pooled leukoreduced platelet concentrates were frozen in a mixture of SSP+, plasma and 6% Me(2)SO. The platelets were concentrated, to avoid post-thaw washing, and frozen at -80°C. The cryopreserved platelet units (n=9) were rapidly thawed at 37°C, reconstituted in 50% SSP+/plasma and stored at 22°C. Platelet recovery and quality were examined 1 and 24h post-thaw and compared to the pre-freeze samples. Upon thawing, platelet recovery ranged from 60% to 80%. However, there were differences between frozen and liquid-stored platelets, including a reduction in aggregation in response to ADP and collagen; increased CD62P expression; decreased viability; increased apoptosis and some loss of mitochondrial membrane integrity. Some recovery of these parameters was detected at 24h post-thaw, indicating an extended shelf-life may be possible. The data suggests that freezing platelets in 6% Me(2)SO and additive solution produces acceptable in vitro platelet quality.

  20. Platelet Lysates Produced from Expired Platelet Concentrates Support Growth and Osteogenic Differentiation of Mesenchymal Stem Cells

    PubMed Central

    Jonsdottir-Buch, Sandra Mjoll; Lieder, Ramona; Sigurjonsson, Olafur Eysteinn

    2013-01-01

    Background Mesenchymal stem cells are promising candidates in regenerative cell therapy. Conventional culture methods involve the use of animal substances, specifically fetal bovine serum as growth supplement. Since the use of animal-derived products is undesirable for human applications, platelet lysates produced from human platelets are an attractive alternative. This is especially true if platelet lysates from already approved transfusion units at blood banks can be utilized. The purpose of this study was to produce human platelet lysates from expired, blood bank-approved platelet concentrates and evaluate their use as growth supplement in the culture of mesenchymal stem cells. Methodology/Principal Findings In this study, bone marrow-derived mesenchymal stem cells were cultured with one of three culture supplements; fetal bovine serum, lysates from freshly prepared human platelet concentrates, or lysates from expired human platelet concentrates. The effects of these platelet-derived culture supplements on basic mesenchymal stem cell characteristics were evaluated. All cultures maintained the typical mesenchymal stem cell surface marker expression, trilineage differentiation potential, and the ability to suppress in vitro immune responses. However, mesenchymal stem cells supplemented with platelet lysates proliferated faster than traditionally cultured cells and increased the expression of the osteogenic marker gene RUNX-2; yet no difference between the use of fresh and expired platelet concentrates was observed. Conclusion/Significance Our findings suggest that human platelet lysates produced from expired platelet concentrates can be used as an alternative to fetal bovine serum for mesenchymal stem cell culture to the same extent as lysates from fresh platelets. PMID:23874839

  1. Expression and function of purinergic receptors in platelets from apheresis-derived platelet concentrates

    PubMed Central

    Koessler, Juergen; Weber, Katja; Koessler, Angela; Yilmaz, Pinar; Boeck, Markus; Kobsar, Anna

    2016-01-01

    Background The storage of platelets affects platelet integrity and functionality, a process named platelet storage lesion (PSL). Reduced adenosine diphosphate (ADP)-induced platelet aggregation is a typical manifestation of PSL. However, the role of ADP receptors in this context has not been evaluated yet. The aim of this study was, therefore, to investigate surface expression and function of the purinergic receptors P2Y1, P2Y12 and P2X1 in stored platelet concentrates. Material and methods Platelets were obtained from venous whole blood and from apheresis-derived platelet concentrates stored for 0, 2 and 5 days. Purinergic receptor expression was measured by flow cytometry and western blot analysis. Receptor function was determined by calcium-induced fluorescence (P2Y1 and P2X1) or by flow cytometric measurement of the platelet reactivity index (P2Y12). Results The basal surface expression and total content of purinergic receptors remained unchanged throughout storage. After an initial reduction during apheresis, P2X1-mediated calcium flux was maintained, whereas the P2Y1-mediated increase of calcium flux gradually decreased during the course of storage. In contrast, the platelet reactivity index was comparable in freshly obtained and stored platelets. Discussion The function of the P2Y12 receptor is maintained during storage of apheresis-derived platelet concentrates. However, the impairment of P2X1 and especially of P2Y1 receptor function indicated by decreased receptor-mediated calcium flux is an important mechanism contributing to reduced ADP responsiveness of stored platelets. PMID:26674810

  2. Evaluation of two instruments for noninvasive platelet concentrate quality assessment.

    PubMed

    George, V; Holme, S; Moroff, G

    1989-01-01

    The Platelet Monitoring System (PMS) and the Non-invasive Assessment of Platelet Shape and Concentration (NAPSAC) instruments which relate light scattering characteristics of platelet concentrates (PC) to platelet concentration and shape, were evaluated to determine their accuracy in assessing platelet quality during storage from 1 to 7 days. The results were correlated with platelet concentration, % discs and pH on 121 PC stored in PL732 containers. NAPSAC output is in the form of platelet concentration and % discs. When NAPSAC and standard method values were compared, correlation coefficients (r) did not exceed 0.76 for counts and 0.62 for % discs. PMS output is in the form of lights with red indicating poor quality and green or amber indicating acceptable quality. Sensitivity of the PMS instrument did not exceed 83% and specificity did not exceed 63%. Mean platelet number, % discs and pH were comparable for units triggering red versus green or amber lights. In a separate study, 13 PL732 PC stored 5 days and transfused autologously were evaluated on the PMS. Three red light units exhibited recovery and survival times similar to those observed with PC triggering green/amber lights. These data indicate that neither instrument adequately assesses the quality of PL732 PC.

  3. Platelet serotonin concentration and depressive symptoms in patients with schizophrenia.

    PubMed

    Peitl, Vjekoslav; Vidrih, Branka; Karlović, Zoran; Getaldić, Biserka; Peitl, Milena; Karlović, Dalibor

    2016-05-30

    Depressive symptoms seem to be frequent in schizophrenia, but so far they have received less attention than other symptom domains. Impaired serotonergic neurotransmission has been implicated in the pathogenesis of depression and schizophrenia. The objectives of this study were to investigate platelet serotonin concentrations in schizophrenic patients with and without depressive symptoms, and to investigate the association between platelet serotonin concentrations and symptoms of schizophrenia, mostly depressive symptoms. A total of 364 patients were included in the study, 237 of which had significant depressive symptoms. Significant depressive symptoms were defined by the cut-off score of 7 or more on Calgary Depression Rating Scale (CDSS). Platelet serotonin concentrations were assessed by the enzyme-linked immunosorbent assay (ELISA). Prevalence of depression in patients with schizophrenia was 65.1%. Schizophrenic patients with depressive symptoms showed lower platelet serotonin concentrations (mean±SD; 490.6±401.2) compared to schizophrenic patients without depressive symptoms (mean±SD; 660.9±471.5). An inverse correlation was established between platelet serotonin concentration and depressive symptoms, with more severe symptoms being associated with lower platelet serotonin concentrations. Depressive symptoms in schizophrenic patients may be associated with reduced concentrations of platelet serotonin.

  4. Ultraviolet irradiation of platelet concentrate abrogates lymphocyte activation without affecting platelet function in vitro

    SciTech Connect

    Kahn, R.A.; Duffy, B.F.; Rodey, G.G.

    1985-11-01

    We studied the effect of ultraviolet (UV) radiation on platelet concentrates. Samples irradiated at 310 mm for 30 minutes at a dose of 1782 J per m2 showed no loss of platelet function in vitro as determined by adenosine diphosphate, collagen, or ristocetin-induced aggregation. Lymphocytes isolated from irradiated units were unable to act as responders or stimulators in a mixed-lymphocyte reaction. These data suggest that UV radiation of platelet concentrates may result in a cell suspension that is unable to evoke an immunological response.

  5. Effective ultraviolet irradiation of platelet concentrates in teflon bags

    SciTech Connect

    Capon, S.M.; Sacher, R.A.; Deeg, H.J. )

    1990-10-01

    Several plastic materials used in blood storage were evaluated for their ability to transmit ultraviolet B (UVB) light. A plastic bag manufactured from sheets of transparent Teflon efficiently (78-86%) transmitted UVB light and was employed in subsequent functional studies of lymphocytes and platelets exposed to UVB light while contained in these bags. In vitro experiments showed a UVB dose-dependent abrogation of lymphocyte responder and stimulator functions, with concurrent preservation of platelet aggregation responses. In a phase I pilot study, UVB-treated platelet concentrates were administered to four bone marrow transplant recipients. Adverse effects attributable to the transfusions were not observed, and patients showed clinically effective transfusion responses. No patient developed lymphocytotoxic HLA or platelet antibodies. These studies suggest that platelets can be effectively irradiated with UVB light in a closed system. However, numerous variables, including container material, volume and composition of contents, steady exposure versus agitation, and exact UV wavelength, must be considered.

  6. Formaldehyde-fixation of platelets for flow cytometric measurement of phosphatidylserine exposure is feasible.

    PubMed

    Rochat, Sophie; Alberio, Lorenzo

    2015-01-01

    Strong platelet activation results in a redistribution of negatively charged phospholipids from the cytosolic to the outer leaflet of the cellular membrane. Annexin V has a high affinity to negatively charged phospholipids and can be used to identify procoagulant platelets. Formaldehyde fixation can cause factitious Annexin V binding. Our aim was to evaluate a method for fixing platelets avoiding additional Annexin V binding. We induced expression of negatively charged phospholipids on the surface of a fraction of platelets by combined activation with convulxin and thrombin in the presence of Annexin V-fluorescein isothiocyanate and calcium. Aliquots of resting and activated platelets were fixed with a low concentration, calcium-free formaldehyde solution. Both native platelets and fixed platelets were analyzed by flow cytometry immediately and after a 24-h storage at 4°C. We observed that the percentage of Annexin V positive resting platelets ranged from 1.5 to 9.3% for the native samples and from 0.4 to 12.8% for the fixed samples (P=0.706, paired t-test). The amount of Annexin V positive convulxin/thrombin activated platelets varied from 12.9 to 35.4% without fixation and from 15.3 to 36.3% after formalin fixation (P=0.450). After a 24-h storage at 4°C, Annexin V positive platelets significantly increased both in the resting and in the convulxin/thrombin activated samples of native platelets (both P<0.001), while results for formalin fixed platelets did not differ from baseline values (P=0.318 for resting fixed platelets; P=0.673 for activated fixed platelets). We conclude that platelet fixation with a low concentration, calcium-free formaldehyde solution does not alter the proportion of Annexin V positive platelets. This method can be used to investigate properties of procoagulant platelets by multicolor flow-cytometric analysis requiring fixation steps.

  7. Factors affecting white cell content in platelet concentrates.

    PubMed

    Champion, A B; Carmen, R A

    1985-01-01

    In this study, we investigated the factors affecting white cell content in platelet concentrates. White cell yields can be reduced 50 percent by stopping platelet-rich plasma expression when the interface is 1 cm from the top of the blood bag as compared to stopping expression when the interface reaches the top of the bag. Further reductions can be achieved by careful handling during transfer of units from the centrifuge cups to expressors (after the first spin) and by carefully balancing units against each other to ensure proper rotor balance during the first spin. Following these suggestions, blood banks should be able to produce platelet concentrates with white cell yields between 2 and 6 X 10(7) and with platelet yields between 7.5 and 8 X 10(10). Transfusion of this product may reduce febrile reactions and lower the incidence of alloimmunizations.

  8. [Risk Assessment of Single-Donor (Apheresis) Platelet Concentrates and Pooled Whole-Blood-Derived Platelet Concentrates].

    PubMed

    Hitzler, Walter; Hutschenreuter, Gabriele; Wartensleben, Herbert

    2015-01-01

    According to the risk estimates of the Robert-Koch-Institute (RKI) and the Paul Ehrlich-Institute (PEI) an equivalence cannot be assumed to exist between the two different platelet preparations. Differences between single-donor (apheresis) platelet concentrates (ATK) and pooled whole-blood-derived platelet concentrates (PTK) result from donor populations, donation intervals, and preparation techniques. There are no prospective randomized studies with regard to the clinical efficacy, which would unambiguously demonstrate equivalence of the therapeutic efficacy of PTK (buffy coat method) in comparison to ATK. The German Association of Blood Transfusion Services (StKB) points out that, due to the non-equivalence of PTK and ATK, it is incumbent on the transfusion physician to select the platelet concentrate, make the appropriate disclosures, and assume treatment responsibility. Proper compensation for ATK and PTK must be ensured by the health insurance companies, whereby a special indication for the selection of either PTK or ATK is not given. Exceptions are patients with known HLA antibodies in which only selected platelet concentrates may be administered. Otherwise, no indication exists in the selection of the different platelet concentrates (Article is in German).

  9. A Comparative Assessment of Quality of Platelet Concentrates Prepared by Buffy Coat Poor Platelet Concentrate Method and Apheresis Derived Platelet Concentrate Method.

    PubMed

    Mallhi, R S; Kumar, Sudeep; Philip, Joseph

    2015-12-01

    Many blood centres in country don't have costly apheresis technology and rely heavily on the platelet production from whole blood donation. We conducted this study with the aim to compare the quality of platelet concentrates (PC) prepared by Buffy Coat derived (BC-PC) and apheresis derived platelet concentrate (Apheresis-PC). Our objective was to collect data by analysis of platelet concentrates prepared by BC-PC methods and Apheresis-PC methods in respect of swirling, volume, platelet count, WBC count and pH of the PC units and elaborate on the quality parameters. Tertiary Care Hospital and Medical College. We assessed a total of 200 BC-PC and 200 Apheresis-PC for their in vitro quality by observing swirling, volume of PC, platelet count/unit, WBC count/unit and pH, to see if they satisfy the recommended quality criteria. Data was analyzed using appropriate statistical technique under the guidance of biostatistician. Apheresis-PC units showed better swirling than BC-PC units (Chi square test; P < 0.05). There was a significant difference in proportion of units satisfying the required volume QC between the two methods (Chi-square test; P < 0.05). Apheresis-PC showed better adherence to the physiological pH values (Student's unpaired t test; P < 0.05). The units of BC-PC and Apheresis-PC did not show significant difference in proportion of units satisfying the Platelet count per unit and residual WBC count per count (Chi square; P 0.203 and 0.617 respectively). There was comparable adherence to QC requirement for platelet count and WBC contamination in two methods. BC-PC were found to be adhering lesser to QC parameters for swirling, volume and pH, but found to be in required QC limits. BCPC can be used effectively in the majority of thrombocytopenic patients in resource poor setting.

  10. The influence of environmental variables on platelet concentration in horse platelet-rich plasma.

    PubMed

    Rinnovati, Riccardo; Romagnoli, Noemi; Gentilini, Fabio; Lambertini, Carlotta; Spadari, Alessandro

    2016-07-04

    Platelet-rich plasma (PRP) commonly refers to blood products which contain a higher platelet (PLT) concentration as compared to normal plasma. Autologous PRP has been shown to be safe and effective in promoting the natural processes of soft tissue healing or reconstruction in humans and horses. Variability in PLT concentration has been observed in practice between PRP preparations from different patients or from the same individual under different conditions. A change in PLT concentration could modify PRP efficacy in routine applications. The aim of this study was to test the influence of environmental, individual and agonistic variables on the PLT concentration of PRP in horses. Six healthy Standardbred mares were exposed to six different variables with a one-week washout period between variables, and PRP was subsequently obtained from each horse. The variables were time of withdrawal during the day (morning/evening), hydration status (overhydration/dehydration) treatment with anti-inflammatory drugs and training periods on a treadmill. The platelet concentration was significantly higher in horses treated with a non-steroidal anti-inflammatory drug (P = 0.03). The leukocyte concentration increased 2-9 fold with respect to whole blood in the PRP which was obtained after exposure to all the variable considered. Environmental variation in platelet concentration should be taken into consideration during PRP preparation.

  11. Concentration of platelets and growth factors in platelet-rich plasma from Goettingen minipigs.

    PubMed

    Jungbluth, Pascal; Grassmann, Jan-Peter; Thelen, Simon; Wild, Michael; Sager, Martin; Windolf, Joachim; Hakimi, Mohssen

    2014-01-01

    In minipigs little is known about the concentration of growth factors in plasma, despite their major role in several patho-physiological processes such as healing of fractures. This prompted us to study the concentration of platelets and selected growth factors in plasma and platelet-rich plasma (PRP) preparation of sixteen Goettingen minipigs. Platelet concentrations increased significantly in PRP in comparison to native blood plasma. Generally, significant increase in the concentration of all growth factors tested was observed in the PRP in comparison to the corresponding plasma or serum. Five of the plasma samples examined contained detectable levels of bone morphogenic protein 2 (BMP-2) whereas eleven of the plasma or serum samples contained minimal amounts of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF-bb) respectively. On the other hand variable concentrations of bone morphogenic protein 7 (BMP-7) and transforming growth factor β1 (TGF-β1) were measured in all plasma samples. In contrast, all PRP samples contained significantly increased amounts of growth factors. The level of BMP-2, BMP-7, TGF-β1, VEGF and PDGF-bb increased by 17.6, 1.5, 7.1, 7.2 and 103.3 fold, in comparison to the corresponding non-enriched preparations. Moreover significant positive correlations were found between platelet count and the concentrations of BMP-2 (r=0.62, p<0.001), TGF-β1 (r=0.85, p<0.001), VEGF (r=0.46, p<0.01) and PDGF-bb (r=0.9, p<0.001). Our results demonstrate that selected growth factors are present in the platelet-rich plasma of minipigs which might thus serve as a source of autologous growth factors.

  12. Concentration of platelets and growth factors in platelet-rich plasma from Goettingen minipigs

    PubMed Central

    Jungbluth, Pascal; Grassmann, Jan-Peter; Thelen, Simon; Wild, Michael; Sager, Martin; Windolf, Joachim; Hakimi, Mohssen

    2014-01-01

    In minipigs little is known about the concentration of growth factors in plasma, despite their major role in several patho-physiological processes such as healing of fractures. This prompted us to study the concentration of platelets and selected growth factors in plasma and platelet-rich plasma (PRP) preparation of sixteen Goettingen minipigs. Platelet concentrations increased significantly in PRP in comparison to native blood plasma. Generally, significant increase in the concentration of all growth factors tested was observed in the PRP in comparison to the corresponding plasma or serum. Five of the plasma samples examined contained detectable levels of bone morphogenic protein 2 (BMP-2) whereas eleven of the plasma or serum samples contained minimal amounts of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF-bb) respectively. On the other hand variable concentrations of bone morphogenic protein 7 (BMP-7) and transforming growth factor β1 (TGF-β1) were measured in all plasma samples. In contrast, all PRP samples contained significantly increased amounts of growth factors. The level of BMP-2, BMP-7, TGF-β1, VEGF and PDGF-bb increased by 17.6, 1.5, 7.1, 7.2 and 103.3 fold, in comparison to the corresponding non-enriched preparations. Moreover significant positive correlations were found between platelet count and the concentrations of BMP-2 (r=0.62, p<0.001), TGF-β1 (r=0.85, p<0.001), VEGF (r=0.46, p<0.01) and PDGF-bb (r=0.9, p<0.001). Our results demonstrate that selected growth factors are present in the platelet-rich plasma of minipigs which might thus serve as a source of autologous growth factors. PMID:26504722

  13. [Pathogen inactivation in platelet concentrates: the French experience].

    PubMed

    Cazenave, J-P

    2011-08-01

    The transfusion of platelet concentrates is increasing in oncohematology patients due to chemotherapy and hematopoietic stem cell grafts. The transmission of pathogenic agents, viruses, parasites and especially bacteria with platelet concentrates stored at room temperature (20-24°C) is associated with a septic risk, partly prevented by bacterial detection. Photochemical inactivation of platelet concentrates, using a technique associating amotosalen and UVA, has been used for five years in a French region for the whole population and a large spectrum of patients, with efficacy and safety. Universal implementation of pathogen inactivation in labile blood products is a major and key step to improve safety against infection in transfusion. Copyright © 2011. Published by Elsevier SAS.

  14. Comparison of growth factor and platelet concentration from commercial platelet-rich plasma separation systems.

    PubMed

    Castillo, Tiffany N; Pouliot, Michael A; Kim, Hyeon Joo; Dragoo, Jason L

    2011-02-01

    Clinical studies claim that platelet-rich plasma (PRP) shortens recovery times because of its high concentration of growth factors that may enhance the tissue repair process. Most of these studies obtained PRP using different separation systems, and few analyzed the content of the PRP used as treatment. This study characterized the composition of single-donor PRP produced by 3 commercially available PRP separation systems. Controlled laboratory study. Five healthy humans donated 100 mL of blood, which was processed to produce PRP using 3 PRP concentration systems (MTF Cascade, Arteriocyte Magellan, Biomet GPS III). Platelet, white blood cell (WBC), red blood cell, and fibrinogen concentrations were analyzed by automated systems in a clinical laboratory, whereas ELISA determined the concentrations of platelet-derived growth factor αβ and ββ (PDGF-αβ, PDGF-ββ), transforming growth factor β1 (TGF-β1), and vascular endothelial growth factor (VEGF). There was no significant difference in mean PRP platelet, red blood cell, active TGF-β1, or fibrinogen concentrations among PRP separation systems. There was a significant difference in platelet capture efficiency. The highest platelet capture efficiency was obtained with Cascade, which was comparable with Magellan but significantly higher than GPS III. There was a significant difference among all systems in the concentrations of WBC, PDGF-αβ, PDGF-ββ, and VEGF. The Cascade system concentrated leukocyte-poor PRP, compared with leukocyte-rich PRP from the GPS III and Magellan systems. The GPS III and Magellan concentrate leukocyte-rich PRP, which results in increased concentrations of WBCs, PDGF-αβ, PDGF-ββ, and VEGF as compared with the leukocyte-poor PRP from Cascade. Overall, there was no significant difference among systems in the platelet concentration, red blood cell, active TGF-β1, or fibrinogen levels. Products from commercially available PRP separation systems produce differing concentrations of

  15. Generation of Platelet Microparticles after Cryopreservation of Apheresis Platelet Concentrates Contributes to Hemostatic Activity.

    PubMed

    Eker, İbrahim; Yılmaz, Soner; Çetinkaya, Rıza Aytaç; Pekel, Aysel; Ünlü, Aytekin; Gürsel, Orhan; Yılmaz, Sebahattin; Avcu, Ferit; Muşabak, Uğur; Pekoğlu, Ahmet; Ertaş, Zerrin; Açıkel, Cengizhan; Zeybek, Nazif; Kürekçi, Ahmet Emin; Avcı, İsmail Yaşar

    2017-03-01

    In the last decade, substantial evidence has accumulated about the use of cryopreserved platelet concentrates, especially in trauma. However, little reference has been made in these studies to the morphological and functional changes of platelets. Recently platelets have been shown to be activated by cryopreservation processes and to undergo procoagulant membrane changes resulting in the generation of platelet-derived microparticles (PMPs), platelet degranulation, and release of platelet-derived growth factors (PDGFs). We assessed the viabilities and the PMP and PDGF levels of cryopreserved platelets, and their relation with thrombin generation. Apheresis platelet concentrates (APCs) from 20 donors were stored for 1 day and cryopreserved with 6% dimethyl sulfoxide. Cryopreserved APCs were kept at -80 °C for 1 day. Thawed APCs (100 mL) were diluted with 20 mL of autologous plasma and specimens were analyzed for viabilities and PMPs by flow cytometry, for thrombin generation by calibrated automated thrombogram, and for PDGFs by enzyme-linked immunosorbent assay testing. The mean PMP and PDGF levels in freeze-thawed APCs were significantly higher (2763±399.4/µL vs. 319.9±80.5/µL, p<0.001 and 550.9±73.6 pg/mL vs. 96.5±49 pg/mL, p<0.001, respectively), but the viability rates were significantly lower (68.2±13.7% vs. 94±7.5%, p<.001) than those of fresh APCs. The mean endogenous thrombin potential (ETP) of freeze-thawed APCs was significantly higher than that of the fresh APCs (3406.1±430.4 nM.min vs. 2757.6±485.7 nM.min, p<0.001). Moreover, there was a significant positive poor correlation between ETP levels and PMP levels (r=0.192, p=0.014). Our results showed that, after cryopreservation, while levels of PMPs were increasing, significantly higher and earlier thrombin formation was occurring in the samples analyzed despite the significant decrease in viability. Considering the damage caused by the freezing process and the scarcity of evidence for their in

  16. Platelet concentration in platelet-rich plasma affects tenocyte behavior in vitro.

    PubMed

    Giusti, Ilaria; D'Ascenzo, Sandra; Mancò, Annalisa; Di Stefano, Gabriella; Di Francesco, Marianna; Rughetti, Anna; Dal Mas, Antonella; Properzi, Gianfranco; Calvisi, Vittorio; Dolo, Vincenza

    2014-01-01

    Since tendon injuries and tendinopathy are a growing problem, sometimes requiring surgery, new strategies that improve conservative therapies are needed. Platelet-rich plasma (PRP) seems to be a good candidate by virtue of its high content of growth factors, most of which are involved in tendon healing. This study aimed to evaluate if different concentrations of platelets in PRP have different effects on the biological features of normal human tenocytes that are usually required during tendon healing. The different platelet concentrations tested (up to 5 × 10(6) plt/µL) stimulated differently tenocytes behavior; intermediate concentrations (0.5 × 10(6), 1 × 10(6) plt/µL) strongly induced all tested processes (proliferation, migration, collagen, and MMPs production) if compared to untreated cells; on the contrary, the highest concentration had inhibitory effects on proliferation and strongly reduced migration abilities and overall collagen production but, at the same time, induced increasing MMP production, which could be counterproductive because excessive proteolysis could impair tendon mechanical stability. Thus, these in vitro data strongly suggest the need for a compromise between extremely high and low platelet concentrations to obtain an optimal global effect when inducing in vivo tendon healing.

  17. Platelet Concentration in Platelet-Rich Plasma Affects Tenocyte Behavior In Vitro

    PubMed Central

    Rughetti, Anna; Dal Mas, Antonella; Properzi, Gianfranco; Calvisi, Vittorio

    2014-01-01

    Since tendon injuries and tendinopathy are a growing problem, sometimes requiring surgery, new strategies that improve conservative therapies are needed. Platelet-rich plasma (PRP) seems to be a good candidate by virtue of its high content of growth factors, most of which are involved in tendon healing. This study aimed to evaluate if different concentrations of platelets in PRP have different effects on the biological features of normal human tenocytes that are usually required during tendon healing. The different platelet concentrations tested (up to 5 × 106 plt/µL) stimulated differently tenocytes behavior; intermediate concentrations (0.5 × 106, 1 × 106 plt/µL) strongly induced all tested processes (proliferation, migration, collagen, and MMPs production) if compared to untreated cells; on the contrary, the highest concentration had inhibitory effects on proliferation and strongly reduced migration abilities and overall collagen production but, at the same time, induced increasing MMP production, which could be counterproductive because excessive proteolysis could impair tendon mechanical stability. Thus, these in vitro data strongly suggest the need for a compromise between extremely high and low platelet concentrations to obtain an optimal global effect when inducing in vivo tendon healing. PMID:25147809

  18. Application of an optimized flow cytometry-based quantification of Platelet Activation (PACT): Monitoring platelet activation in platelet concentrates

    PubMed Central

    Roest, Mark; Henskens, Yvonne M. C.; de Laat, Bas; Huskens, Dana

    2017-01-01

    Background Previous studies have shown that flow cytometry is a reliable test to quantify platelet function in stored platelet concentrates (PC). It is thought that flow cytometry is laborious and hence expensive. We have optimized the flow cytometry-based quantification of agonist induced platelet activation (PACT) to a labor, time and more cost-efficient test. Currently the quality of PCs is only monitored by visual inspection, because available assays are unreliable or too laborious for use in a clinical transfusion laboratory. Therefore, the PACT was applied to monitor PC activation during storage. Study design and methods The optimized PACT was used to monitor 5 PCs during 10 days of storage. In brief, optimized PACT uses a ready-to-use reaction mix, which is stable at -20°C. When needed, a test strip is thawed and platelet activation is initiated by mixing PC with PACT. PACT was based on the following agonists: adenosine diphosphate (ADP), collagen-related peptide (CRP) and thrombin receptor-activating peptide (TRAP-6). Platelet activation was measured as P-selectin expression. Light transmission aggregometry (LTA) was performed as a reference. Results Both PACT and LTA showed platelet function decline during 10-day storage after stimulation with ADP and collagen/CRP; furthermore, PACT showed decreasing TRAP-induced activation. Major differences between the two tests are that PACT is able to measure the status of platelets in the absence of agonists, and it can differentiate between the number of activated platelets and the amount of activation, whereas LTA only measures aggregation in response to an agonist. Also, PACT is more time-efficient compared to LTA and allows high-throughput analysis. Conclusion PACT is an optimized platelet function test that can be used to monitor the activation of PCs. PACT has the same accuracy as LTA with regard to monitoring PCs, but it is superior to both LTA and conventional flow cytometry based tests with regard to labor

  19. Leukoreduction of platelet concentrates using a 'polishing' filter.

    PubMed

    van der Meer, P F; Pietersz, R N; Reesink, H W

    2000-01-01

    Filters for removal of leukocytes from platelet concentrates (PCs) usually have a large volume to guarantee sufficient leukoreduction. In this study, a small filter, with a volume of only 8 ml and therefore minimal platelet loss, for leukoreduction of PCs was investigated. This filter has a 'limited' leukoreducing capacity, hence the filter is called a 'polishing' filter. PCs were made from 5 pooled buffy coats in either plasma or additive solution (PAS-II). After centrifugation, the platelet-rich supernatant was expressed on an automated separator (Compomat G4) to an empty transfer bag. The content of this transfer bag was filtered into the platelet storage bag, either by expression by lowering the top press of the Compomat G4, or by gravity by hanging it on a filtration rack. Leukocyte counts before and after filtration revealed a mean leukoreducing capacity for the filter of 2.67 log(10) and a platelet loss of only 2% for PCs in PAS-II (n = 50), and for PCs in plasma a 3.43 log(10) leukoreduction with 3% platelet loss (n = 30). Expression of the PCs both in plasma and PAS-II through the filter using the Compomat G4 resulted in 10/10 units containing <5x10(6) leukocytes, but 1/10 PCs contained >1x10(6) leukocytes for both solutions. Filtration by gravity resultet in 40/40 units with <1x10(6) leukocytes for PCs in plasma, and 60/60 units with <1x10(6) for PCs in PAS-II. The 'polishing' filter allows reliable, standardized and automated production of PCs, both in plasma and additive solution with minimal platelet loss, and containing uniformly <1x10(6) leukocytes, provided the filtration procedure is performed by gravity.

  20. Increasing platelet concentrations in leukocyte-reduced platelet-rich plasma decrease collagen gene synthesis in tendons.

    PubMed

    Boswell, Stacie G; Schnabel, Lauren V; Mohammed, Hussni O; Sundman, Emily A; Minas, Tom; Fortier, Lisa A

    2014-01-01

    Platelet-rich plasma (PRP) is used for the treatment of tendinopathy. There are numerous PRP preparations, and the optimal combination of platelets and leukocytes is not known. Within leukocyte-reduced PRP (lrPRP), there is a plateau effect of platelet concentration, with increasing platelet concentrations being detrimental to extracellular matrix synthesis. Controlled laboratory study. Different formulations of lrPRP with respect to the platelet:leukocyte ratio were generated from venous blood of 8 horses. Explants of the superficial digital flexor tendon were cultured in lrPRP products for 96 hours. Platelet-derived growth factor-BB (PDGF-BB), tumor necrosis factor-α (TNF-α), transforming growth factor-β1 (TGF-β1), and interleukin-1β (IL-1β) concentrations were determined in the media by enzyme-linked immunosorbent assay. Gene expression in tendon tissue for collagen type I and III (COL1A1 and COL3A1, respectively), matrix metalloproteinase-3 and -13 (MMP-3 and MMP-13, respectively), cartilage oligomeric matrix protein (COMP), and IL-1β was determined. Data were divided into 3 groups of lrPRP based on the ratio of platelets:leukocytes and evaluated to determine the effect of platelet concentration. Complete blood counts verified leukocyte reduction and platelet enrichment in all PRP preparations. In the lrPRP preparation, the anabolic growth factors PDGF-BB and TGF-β1 were increased with increasing platelet concentrations, and the catabolic cytokine IL-1β was decreased with increasing platelet concentrations. Increasing the platelet concentration resulted in a significant reduction in COL1A1 and COL3A1 synthesis in tendons. Increasing the platelet concentration within lrPRP preparations results in the delivery of more anabolic growth factors and less proinflammatory cytokines, but the biological effect on tendons is diminished metabolism as indicated by a decrease in the synthesis of both COL1A1 and COL3A1. Together, this information suggests that

  1. Overcoming the bottleneck of platelet lysate supply in large-scale clinical expansion of adipose-derived stem cells: A comparison of fresh versus three types of platelet lysates from outdated buffy coat-derived platelet concentrates.

    PubMed

    Glovinski, Peter V; Herly, Mikkel; Mathiasen, Anders B; Svalgaard, Jesper D; Borup, Rehannah; Talman, Maj-Lis M; Elberg, Jens J; Kølle, Stig-Frederik T; Drzewiecki, Krzysztof T; Fischer-Nielsen, Anne

    2017-02-01

    Platelet lysates (PL) represent a promising replacement for xenogenic growth supplement for adipose-derived stem cell (ASC) expansions. However, fresh platelets from human blood donors are not clinically feasible for large-scale cell expansion based on their limited supply. Therefore, we tested PLs prepared via three methods from outdated buffy coat-derived platelet concentrates (PCs) to establish an efficient and feasible expansion of ASCs for clinical use. PLs were prepared by the freeze-thaw method from freshly drawn platelets or from outdated buffy coat-derived PCs stored in the platelet additive solution, InterSol. Three types of PLs were prepared from outdated PCs with platelets suspended in either (1) InterSol (not manipulated), (2) InterSol + supplemented with plasma or (3) plasma alone (InterSol removed). Using these PLs, we compared ASC population doubling time, cell yield, differentiation potential and cell surface markers. Gene expression profiles were analyzed using microarray assays, and growth factor concentrations in the cell culture medium were measured using enzyme-linked immunosorbent assay (ELISA). Of the three PL compositions produced from outdated PCs, removal of Intersol and resuspension in plasma prior to the first freezing process was overall the best. This specific outdated PL induced ASC growth kinetics, surface markers, plastic adherence and differentiation potentials comparable with PL from fresh platelets. ASCs expanded in PL from fresh versus outdated PCs exhibited different expressions of 17 overlapping genes, of which 10 were involved in cellular proliferation, although not significantly reflected by cell growth. Only minor differences in growth factor turnover were observed. PLs from outdated platelets may be an efficient and reliable source of human growth supplement allowing for large-scale ASC expansion for clinical use. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights

  2. Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF).

    PubMed

    Dohan Ehrenfest, David M; Rasmusson, Lars; Albrektsson, Tomas

    2009-03-01

    The topical use of platelet concentrates is recent and its efficiency remains controversial. Several techniques for platelet concentrates are available; however, their applications have been confusing because each method leads to a different product with different biology and potential uses. Here, we present classification of the different platelet concentrates into four categories, depending on their leucocyte and fibrin content: pure platelet-rich plasma (P-PRP), such as cell separator PRP, Vivostat PRF or Anitua's PRGF; leucocyte- and platelet-rich plasma (L-PRP), such as Curasan, Regen, Plateltex, SmartPReP, PCCS, Magellan or GPS PRP; pure plaletet-rich fibrin (P-PRF), such as Fibrinet; and leucocyte- and platelet-rich fibrin (L-PRF), such as Choukroun's PRF. This classification should help to elucidate successes and failures that have occurred so far, as well as providing an objective approach for the further development of these techniques.

  3. Applications of ultraviolet light in the preparation of platelet concentrates

    SciTech Connect

    Pamphilon, D.H.; Corbin, S.A.; Saunders, J.; Tandy, N.P.

    1989-06-01

    Passenger lymphocytes in platelet concentrates (PCs) may induce the formation of lymphocytotoxic antibodies (LCTAbs) and subsequent refractoriness to platelet transfusions. Ultraviolet (UV) irradiation can prevent lymphocytes' acting as stimulator or responder cells in mixed-lymphocyte reactions (MLRs) and could theoretically prevent LCTAb formation in vivo. A system has been devised for the delivery of UV irradiation to PCs; platelet storage characteristics and MLRs were evaluated in UV-irradiated PCs harvested from healthy donors with the Haemonetics V50 and PCS cell separators. MLR and response to phytohemagglutinin stimulation were abolished by a dose of 3000 joules per m2 at a mean wavelength of 310 nm. Platelet aggregatory responses to adenosine diphosphate (ADP), ristocetin, collagen and epinephrine, hypotonic shock response, and pH showed no important differences when control PCs and PCs irradiated as above were compared during 5 days of storage in Fenwal PL-1240 packs. Lactate production during storage was significantly higher in UV-treated PCs (p less than 0.001), but values did not exceed 20 mmol per L. UV transmission at 310 nm in standard blood product containers, including the Fenwal PL-146, PL-1240, and PL-732, was low (less than 30%), but it was acceptable in the Delmed Cryostorage and DuPont SteriCell packs (greater than 50%). UV irradiation may provide a simple and inexpensive means of producing nonimmunogenic PCs.

  4. Platelet concentrates transfusion in cardiac surgery in relation to preoperative point-of-care assessment of platelet adhesion and aggregation.

    PubMed

    Solomon, Cristina; Hartmann, Jennifer; Osthaus, Alexander; Schöchl, Herbert; Raymondos, Kostas; Koppert, Wolfgang; Rahe-Meyer, Niels

    2010-01-01

    Platelet dysfunction is an important cause of bleeding early after cardiac surgery. Whole-blood multiple electrode aggregometry (MEA), investigating the adhesion and aggregation of activated platelets onto metal electrodes, has shown correlations with platelet concentrates transfusion in this setting. Platelet activity in vivo is dependent on shear stress, an aspect that cannot be investigated with MEA, but with the cone and plate(let) analyzer (CPA) Impact-R that measures the interaction of platelets and von Willebrand factor (vWF) in whole blood under shear. We hypothesized that preoperative CPA may show better correlation with platelet concentrates transfusion post-cardiac surgery than MEA, since it is dependent on both platelet activity and platelet interaction with vWF multimers. Blood was obtained preoperatively from 30 patients undergoing aorto-coronary bypass (ACB) and 20 patients with aortic valve (AV) surgery. MEA was performed in hirudin-anticoagulated blood. The Impact-R analyses were performed in blood anticoagulated with hirudin, heparin or the standard anticoagulant citrate. For the light microscopy images obtained, the parameter surface coverage (SC) was calculated. Preoperative Impact-R results were abnormally decreased in AV patients and significantly lower than in ACB patients. For the Impact-R analysis performed in citrated blood, no correlation with platelet concentrates transfusion was observed. In contrast, MEA was comparable between the groups and correlated significantly with intraoperative platelet concentrates transfusion in both groups (rho between -0.47 and -0.62, p < 0.05). Multiple electrode aggregometry appeared more useful and easier to apply than CPA for preoperatively identifying patients with platelet concentrates transfusion in cardiac surgery.

  5. Buffy-coat-derived pooled platelet concentrates and apheresis platelet concentrates: which product type should be preferred?

    PubMed

    Schrezenmeier, H; Seifried, E

    2010-07-01

    There is an ongoing debate whether platelet concentrates (PCs) prepared from either whole-blood donations or by plateletpheresis are superior. Usage of these two product types varies greatly between countries and individual institutions. Some use mainly apheresis PCs; others prefer pooled PCs which are produced from whole-blood donations. This review summarizes the existing information on these product types. In the first part data on quality, efficacy and safety are reviewed. It is important to note that the issue cannot be answered just by comparing 'the' apheresis platelet concentrate versus 'the' pooled platelet concentrate. Other factors which determine the quality of a product, e.g. residual leukocyte count, plasma content, additive solution or storage period may be even more important. The focus of the debate should be shifted. It is much more needed to further improve the overall quality of PCs and to optimize treatment of thrombocytopenic patients than to concentrate on a single-edged view on just the preparation method. In the second part of this review we compare the product types from the donor's point of view. If PCs which are equally safe and effective can be obtained by various methods, ethics and the safety of the healthy volunteer donor tips the scales. The decision on the use of a particular product type should take into account all aspects of efficacy, side effects and availability of the product as well as the donor's perspective and the commitment to maximize the use of the valuable whole-blood donation.

  6. Modified CMI, an essential adjunct to CMI of platelet for quality control during preparation and storage of platelet concentrates.

    PubMed

    Ray, Vijayalaxmi; Chaudhary, Rajendra; Singh, Harprit

    2003-10-01

    Changes in platelet indices such as platelet count (PLT), mean platelet volume (MPV) and corrected morphological index (CMI) have been correlated with the quality of platelet concentrates during storage but the acceptability criteria for these is not clearly defined. Platelet distribution width (PDW) is a measure of platelet anisocytosis and together with the MPV provides an adjunctive measure of quality. Hence, we investigated the use of the MPV and PDW with and without EDTA in an assessment of the quality of stored platelet concentrates. The differences in platelet count (dPLT), in MPV (dMPV) and in PDW (dPDW) with and without EDTA incubation were calculated. CMI and the modified CMI were further derived using dPLT, dMPV, and dPDW by simple mathematical calculations. We observed a good correlation of the CMI and modified CMI (r=0.966, p<0.01). The dPDWs underwent significant changes during the storage period in addition to changes in dPLT and dMPV. The dPDW provides a test of good power when compared with the dMPV (R(2)=64.1 and 26.1 respectively). We have observed an increment of 92.76% in the dPDW while it was 71% in the dMPV. This clearly shows that the dPDW is a better marker of quality control when compared to the dMPV during processing and storage of platelet concentrates.

  7. [Introduction of platelet additive solution in platelet concentrates: towards a decrease of blood transfusion reactions].

    PubMed

    Rebibo, D; Simonet, M; Hauser, L

    2008-11-01

    Platelet concentrates (PC) are used in thrombocytopenia for curative or preventive treatment for hemorrhagic risk. Since five years, additive solutions have been added in PCs for several reasons; one of them is to present an interest in the intolerance in plasma reactions. The literature data have shown that these solutions entail fewer allergic reactions than PCs kept in plasma. This study was reviewed on three years of transfusion in France. The main objective of this study was to see if there was a difference in frequency when these PCs were in solution or not. All adverse reactions in recipients (ARR) occurring among PCs recipients (with and without additive solution) were analysed. The categories of ARR specifically studied were: allergies, febril non haemolytic reactions (FNHR) and the category "unknown". This study shows that there is significantly lower incidence of allergies by introducing solution. For all ARRs, there is also a decrease in their frequency when PCs are in additive solution, it is significant except for the apheresis platelet concentrates. For categories FNHR and "unknown", the results are opposed and/or not significant. This study confirms that introduction of additive solutions in PCs is able to reduce some allergic transfusion reactions.

  8. Sterility screening of platelet concentrates: questioning the optimal test strategy.

    PubMed

    Dreier, J; Störmer, M; Pichl, L; Schottstedt, V; Grolle, A; Bux, J; Kleesiek, K

    2008-10-01

    Routine bacterial monitoring of apheresis platelet concentrates (APC) and pooled platelet concentrates (PPC) was introduced in two German blood services using culture and real-time reverse transcriptase (RT)-polymerase chain reaction (PCR). The results of testing are reviewed and used to discuss different strategies for detection of bacterial contamination of PCs. Two thousand three hundred and sixty-two APCs and 1993 PPCs have been tested by real-time RT-PCR and the BacT/Alert automated culturing system using aerobic and anaerobic culture bottles. After standard processing of PCs and storage of 22-24 h at 20-24 degrees C with agitation, samples were taken under aseptic conditions. Reactive culture bottles were confirmed as positive and bacterial isolates were identified by 16S rRNA analysis and biochemical tests. Seventeen of 2362 tested APCs were reactive in culture and one also in RT-PCR. Of these, 13 APCs were identified as initially positive as Staphylococcus warneri (n = 1, positive in aerobic and anaerobic culture), Propionibacterium acnes (n = 12, positive only in anaerobic culture) and four were initially reactive. Two of 1993 PPCs were initially reactive (anaerobic) and two more were confirmed positive (anaerobic) from a repeat culture and identified as P. acnes. All remaining specimens were tested negative. Our study demonstrates that the predominant organisms implicated in platelet bacterial contamination are part of the human skin flora. Inoculating blood culture systems and anaerobic cultivation detects these bacteria after approximately 3-7 days when blood products have been transfused. Based on the presented data different screening strategies are discussed.

  9. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part I: technological concepts and evolution.

    PubMed

    Dohan, David M; Choukroun, Joseph; Diss, Antoine; Dohan, Steve L; Dohan, Anthony J J; Mouhyi, Jaafar; Gogly, Bruno

    2006-03-01

    Platelet-rich fibrin (PRF) belongs to a new generation of platelet concentrates geared to simplified preparation without biochemical blood handling. In this initial article, we describe the conceptual and technical evolution from fibrin glues to platelet concentrates. This retrospective analysis is necessary for the understanding of fibrin technologies and the evaluation of the biochemical properties of 3 generations of surgical additives, respectively fibrin adhesives, concentrated platelet-rich plasma (cPRP) and PRF. Indeed, the 3-dimensional fibrin architecture is deeply dependent on artificial clinical polymerization processes, such as massive bovine thrombin addition. Currently, the slow polymerization during PRF preparation seems to generate a fibrin network very similar to the natural one. Such a network leads to a more efficient cell migration and proliferation and thus cicatrization.

  10. [Anti-A antibodies and bacterial contamination of platelet concentrates].

    PubMed

    García-Erce, J A; Seoane, A; Solano, V M; Salvador Osuna, C; Pérez-Layo, A; Gómez-Arteta, E; Gimeno, J J

    1999-12-01

    The possible ABO group antibodies protective function against several infections has been classically described. We analyze the platelet concentrates (PC) bacterial control results and their ABO antibodies. We studied 245 outdated PCs (> 5 days). The samples were sterilely collected for adequate microbiological investigation studies on sheep-blood agar plates. If bacterial growth is found, the microbiological identification is performed on the basis of standard tests, the specific anti-biotype being achieved by disk-diffusion method on Müeller-Hinton agar plates, and the red cell concentrate was analyzed. Bacterial growth by negative coagulase Staphylococcus was found in 10 PCs (4.1%; CI95%; 1.97-7.37). The contaminated PCs lacked natural anti-A antibodies. There were no statistical differences when we analyzed the PC's age, colour or blood group. The anti-A antibodies may be a protective factor versus PCs contamination caused by resident bacteria.

  11. Propionibacterium acnes lacks the capability to proliferate in platelet concentrates.

    PubMed

    Störmer, M; Kleesiek, K; Dreier, J

    2008-04-01

    Propionibacterium acnes is considered to be one of the most frequent contaminants of platelet concentrates (PCs) when anaerobic culture-based detection methods are used. But Propionibacteria are often detected too late when blood products have already been transfused. Therefore, its transfusion relevance is still demanding clarification because studies of the outcome of patients transfused with P. acnes-contaminated PCs are still uncommon. In this study, we monitored clinical effects in patients after transfusion of PCs, which were detected too late in sterility testing. Furthermore, we assessed the bacterial proliferation of Propionibacterium species seeded into PCs to clarify their significance for platelet bacteria screening. In the look-back process, we followed the route of the putative contaminated PC units from storage to transfusion. In the in vitro study, PCs were inoculated with 1-100 colony-forming unit (CFU)/ml of clinical isolates of Propionibacteria (n = 10). Sampling was performed during 10-day aerobic storage at 22 degrees C. The presence of bacteria was assessed by plating culture and automated BacT/Alert culture system. Propionibacterium acnes shows slow or no growth under PC storage conditions. Clinical signs of adverse events after transfusion of potentially contaminated PC units were not reported. Propionibacteria do not proliferate under PC storage conditions and therefore may be missed or detected too late when blood products have already been transfused.

  12. Platelets

    MedlinePlus

    ... common disorder of platelet function is caused by aspirin. Aspirin blocks one of the steps required for platelets to stick together. This effect of aspirin is what makes it an effective treatment for ...

  13. Increasing platelet concentration in platelet-rich plasma inhibits anterior cruciate ligament cell function in three-dimensional culture.

    PubMed

    Yoshida, Ryu; Cheng, Mingyu; Murray, Martha M

    2014-02-01

    Tissue engineering is one new strategy being developed to treat ACL ruptures. One such approach is bio-enhanced ACL repair, where a suture repair is supplemented with a bio-active scaffold containing platelets. However, the optimal concentration of platelets to stimulate ACL healing is not known. We hypothesized that increasing platelet concentrations in the scaffold would enhance critical cell behaviors. Porcine ACL fibroblasts were obtained from explant culture and suspended in platelet poor plasma (PPP), 1× platelet-rich plasma (PRP), 3× PRP, 5× PRP, or phosphate buffered saline (PBS). The cell suspensions were cultured in a 3D collagen scaffold. Cellular metabolism (MTT assay), apoptosis (TUNEL assay), and gene expression for type I and type III collagen were measured. 1× PRP significantly outperformed 5× PRP in all parameters studied: Type I and III collagen gene expression, apoptosis prevention, and cell metabolism stimulation. ACL fibroblasts cultured with 1× PRP had the highest type I and type III collagen gene expression. 1× PRP and PPP groups had the highest cell metabolism and lowest apoptosis rates. Concentration of platelets had significant effects on the behavior of ACL fibroblasts; thus, it is an important parameter that should be specified in clinical or basic science studies.

  14. Use of 8-methoxypsoralen and long-wavelength ultraviolet radiation for decontamination of platelet concentrates

    SciTech Connect

    Lin, L.; Wiesehahn, G.P.; Morel, P.A.; Corash, L. )

    1989-07-01

    Transmission of viral diseases through blood products remains an unsolved problem in transfusion medicine. We have developed a psoralen photochemical system for decontamination of platelet concentrates in which platelets are treated with long wavelength ultraviolet radiation (UVA, 320-400 nm) in the presence of 8-methoxypsoralen (8-MOP). Bacteria, RNA viruses, and DNA viruses ranging in genome size from 1.2 x 10(6) daltons, encompassing the size range of human pathogens, were inoculated into platelet concentrates and subjected to treatment. This system inactivated 25 to 30 logs/h of bacteria Escherichia coli or Staphylococcus aureus, 6 logs/h of bacteriophage fd, 0.9 log/h of bacteriophage R17 and 1.1 logs/h of feline leukemia virus (FeLV) in platelet concentrates maintained in standard storage bags. Platelet integrity and in vitro function before, immediately following photochemical treatment, and during prolonged storage after treatment, were evaluated by measuring: (1) extracellular pH; (2) platelet yields; (3) extracellular lactate dehydrogenase (LDH) levels; (4) platelet morphology; (5) platelet aggregation responsiveness; (6) thromboxane beta-2 (TXB-2) production; (7) dense body secretion; and (8) alpha granule secretion. These assays demonstrated that this photochemical inactivation system inactivated bacteria and viruses in platelet concentrates with minimal adverse effects on the in vitro function of platelets in comparison to untreated control concentrates maintained under current, standard blood bank conditions.

  15. [Qualitative comparison between buffy-coat-collected platelet concentrates and those by single-donor plateletpheresis].

    PubMed

    Yu, Yang; Luo, Qun; Liu, Jin-Han

    2007-08-01

    This study was aimed to compare the difference of quality between buffy-coat-collected platelet concentrates (BC-PC) and single-donor plateletpheresis (SDP). 15 packs of BC-PC and 15 units SDP were stored at 20 degrees C - 24 degrees C with agitation. Platelet concentration, platelet volume, residual leukocyte and residual erythrocyte in two groups were examined after preparation for 1 hour. Mean platelet volume, pH value, hypotonic shock response (HSR), CD62p expression and CD62p re-expression of platelet were detected on 0, 1, 2, 3, 4, 5 days of platelet preservation. The results showed that the platelet yields, residual leukocyte and residual erythrocyte in two groups accorded with the national quality standard respectively, but residual leukocyte and residual erythrocyte in BC-PC group were higher than those in SDP group when platelet yields in two groups were equal (p < 0.01). Lactate concentration, CD62p expression of platelet increased with prolongation of preseved time, while pH value decreased gradually. Compared with SDP group, there were significant differences in CD62p expression, CD62p re-expression of platelet preserved for 0 - 5 days (p < 0.01), and in pH value of platelet preserved 2 - 5 days (p < 0.01). There was no changes in HSR of SDP group for 0 - 5 days, while HSR in BC-PC group decreased gradually. There were significant differences in HSR of platelet preserved for 1 - 5 days (p < 0.01). It is concluded that the platelet concentrates prepared by BC-PC are not equal to SDP in quality, the preparation technology of BC-PC should be optimized further in order to reduce residual leukocyte, residual erythrocyte and activated platelet yields, as well as improve the quality of BC-PC.

  16. Expired and Pathogen-Inactivated Platelet Concentrates Support Differentiation and Immunomodulation of Mesenchymal Stromal Cells in Culture.

    PubMed

    Jonsdottir-Buch, Sandra Mjoll; Sigurgrimsdottir, Hildur; Lieder, Ramona; Sigurjonsson, Olafur Eysteinn

    2015-01-01

    Platelet lysates have been reported as suitable cell culture supplement for cultures of mesenchymal stromal cells (MSCs). The demand for safe and animal-free cultures of MSCs is linked to the potential application of MSCs in clinics. While the use of platelet lysates offers an alternative to animal serum in MSC cultures, obtaining supplies of fresh platelet concentrates for lysate production is challenging and raises concerns due to the already existing shortage of platelet donors. We have previously demonstrated that expired platelet concentrates may represent a good source of platelets for lysate production without competing with blood banks for platelet donors. The INTERCEPT Blood System™ treatment of platelet concentrates allows for prolonged storage up to 7 days, using highly specific technology based on amotosalen and UV-A light. The INTERCEPT system has therefore been implemented in blood processing facilities worldwide. In this study, we evaluated the suitability of INTERCEPT-treated, expired platelet concentrates, processed into platelet lysates, for the culture of MSCs compared to nontreated expired platelets. Bone marrow-derived MSCs were cultured in media supplemented with either platelet lysates from traditionally prepared expired platelet concentrates or in platelet lysates from expired and pathogen-inactivated platelet concentrates. The effects of pathogen inactivation on the ability of the platelets to support MSCs in culture were determined by evaluating MSC immunomodulation, immunophenotype, proliferation, and trilineage differentiation. Platelet lysates prepared from expired and pathogen-inactivated platelet concentrates supported MSC differentiation and immunosuppression better compared to traditionally prepared platelet lysates from expired platelet units. Pathogen inactivation of platelets with the INTERCEPT system prior to use in MSC culture had no negative effects on MSC immunophenotype or proliferation. In conclusion, the use of expired

  17. Evaluation of autologous platelet concentrate for intertransverse process lumbar fusion.

    PubMed

    Sethi, Paul M; Miranda, Jose J; Kadiyala, Sudha; Patel, Tushar Ch; Panjabi, Manohar; Troiano, Nancy; Friedlaender, Gary E

    2008-04-01

    Data on the role of platelet concentrate (PC) in spinal fusion are limited. Using the New Zealand white rabbit model, we compared fusion rates at L5-L6 using 2 different volumes (1.5 cm(3), 3.0 cm(3)) of iliac crest autograft with and without PC (4 groups total, 10 animals in each). PC was collected from donor rabbits and adjusted to a concentration of 1 x 10(6) platelets/mL. Bone growth and fusion were evaluated using biomechanical, radiographic, and histologic testing. At 1.5 cm(3), autograft alone had a 29% fusion rate, compared with autograft plus PC, which had a 57% fusion rate (P = .06). At 3.0 cm(3), the fusion rate approached 90% in both groups. Radiologic fusion had a 70% correlation with biomechanical test results. Huo/Friedlaender scores were 4.3 (SD, 2.9) for 1.5-cm(3) autograft alone; 5.0 (SD, 3.5) for 1.5-cm(3) autograft plus PC; 4.7 (SD, 2.5) for 3.0-cm(3) autograft alone; and 7.7 (SD, 0.6) for 3.0-cm(3) autograft plus PC. For 1.5-cm(3) autograft, a trend toward improvement in biomechanically defined fusion was found when PC was added, which suggests that, when the amount of bone graft is limited, PC may function as a graft extender in posterolateral fusion. At higher volumes of bone graft, no appreciable difference was noted between groups. Although radiography revealed fusion masses, the technique was not useful in identifying pseudarthrosis. On histologic analysis, adding PC seemed to result in more mature bone at both volumes, with the most mature bone in the group with 3.0-cm(3) autograft plus PC.

  18. Normal platelet function in platelet concentrates requires non-platelet cells: a comparative in vitro evaluation of leucocyte-rich (type 1a) and leucocyte-poor (type 3b) platelet concentrates

    PubMed Central

    Parrish, William R; Roides, Breana; Hwang, Julia; Mafilios, Michael; Story, Brooks; Bhattacharyya, Samir

    2016-01-01

    Background Therapeutic success of platelet-rich plasma (PRP) may vary based on the composition and preparation method. The objective of this study was to evaluate the cellular components of platelet concentrates produced by a leucocyte-rich (LR-PRP) and a leucocyte-poor PRP systems (LP-PRP). Methods Parameters evaluated included platelet recovery, platelet concentration, red blood cell (RBC) and white blood cell (WBC) composition, platelet growth factor release and stimulation of human tendon cell proliferation in vitro. Results Platelet recoveries were 52% for LP-PRP and 89% for LR-PRP. LR-PRP demonstrated greater reproducibility with a 4.2% coefficient of variation (CV) compared with 19.4% for LP-PRP (p<0.001). LR-PRP demonstrated a greater increase in platelet concentration (7.9-fold) than LP-PRP (2.2-fold; p<0.001). LP-PRP showed 5.0-fold reductions in WBCs, while LR-PRP showed a 4.0-fold increase (p<0.001). LP-PRP reduced RBCs to a haematocrit of 0.25, while LR-PRP reduced haematocrit to 11.8. LP-PRP did not coagulate robustly on reactivation with CaCl2, and released significantly lower levels of epidermal growth factor (EGF) and transforming growth factor β1 (TGF-β1) than whole blood (p<0.03). LP-PRP also did not stimulate tendon cell proliferation greater than whole blood. In contrast, LR-PRP showed increases in each growth factor on activation with CaCl2 (p<0.01) and stimulated greater proliferation (p<0.05) compared with whole blood. Forced activation of LP-PRP with exogenous thrombin rescued the coagulation deficiency and induced greater growth factor release than comparable whole blood (p<0.03). Conclusions These data suggest that non-platelet cellular components in platelet concentrates are important for proper platelet function, including thrombin generation, growth factor release and clot retraction. PMID:27900155

  19. Counting platelets in platelet concentrates on hematology analyzers: a multicenter comparative study.

    PubMed

    van der Meer, Pieter F; Dijkstra-Tiekstra, Margriet J; Mahon, Anne; de Wildt-Eggen, Janny

    2009-01-01

    Hematology analyzers are designed to count whole blood samples, but are also used by blood centers to perform quality control on blood components. In platelet (PLT) concentrates, the number of PLTs is approximately fivefold higher and red blood cells are absent, causing variable PLT counting results. It was our aim to compare currently used hematology analyzers for counting PLTs in PLT concentrates using fixed human PLTs. PLT samples were fixed, diluted into seven concentration levels (plus one blank), aliquoted, and shipped to 68 centers. Evaluable data were obtained for 89 hematology analyzers. All samples were counted six times, and results were reported to the coordinating center. The overall group mean was calculated, and the percentage deviation from this mean was calculated for each analyzer. At PLT levels relevant for blood centers, 750 x 10(9) to 2000 x 10(9) per L, analyzers gave results that were between 35 percent lower and 16 percent higher than the overall group mean. Within a group of analyzers, results were comparable with coefficient of variations usually below 10 percent, indicating that the observed differences were caused by instrument characteristics. A smaller study with fresh, unfixed PLT samples showed that analyzers behaved similarly for fixed and fresh PLTs. With a wide array of currently used hematology analyzers, a marked difference was determined for the PLT counts of fixed human-based identical samples provided to 68 laboratories by a centralized facility. A gold standard method is needed to allow for more valid interlaboratory comparisons between hematology analyzers.

  20. Monitoring of Platelet Activation in Platelet Concentrates Using Transmission Electron Microscopy

    PubMed Central

    Neumüller, Josef; Meisslitzer-Ruppitsch, Claudia; Ellinger, Adolf; Pavelka, Margit; Jungbauer, Christof; Renz, Renate; Leitner, Gerda; Wagner, Thomas

    2013-01-01

    Summary Objective The quality of platelet concentrates (PC) is important for the in vivo recovery of thrombostasis in patients suffering from bleeding disorders and in tumor patients after chemotherapy. In this respect, activated platelets (PLT) cannot display their full functionality in the recipient and even can cause adverse effects. Therefore, we developed a transmission electron microscopy (TEM) method for quality assessment of PC. Methods Score values taken from panorama TEM images describe the progress of PLT activation. To exemplify this method, i) 19 apheresis PC isolated with the Baxter Amicus system (BA) were compared with 14 PC obtained from pooled buffy coats (BC). ii) The score values of 33 PC derived from BA as well from BC were compared with flow-cytometric CD62P determinations by cross correlation. iii) Changes in the score value profiles during storage of a single pathogen-reduced BA PC were monitored over a period of 7 days. Results The TEM evaluation described allows for demonstrating particular PLT activation stages. i) Significant differences between the percentages of the score values 0, 1 and 2 could be demonstrated in both processing groups. No significant differences were found comparing these two groups. ii) A weak correlation could be shown when comparing the percentages of score values 2 plus 3 with the percentage of CD62P-positive PLT. iii) The pathogen reduction affected slightly the score profiles during storage due to an increase of dead PLT. Conclusion Our investigations demonstrate the unique detailed quality information of PC obtained by the TEM method. This method can be performed in every routine electron microscopy laboratory. PMID:23652838

  1. Quality assessment and transfusion efficacy of buffy coat-derived platelet concentrates washed with platelet additive solution.

    PubMed

    Fernández-Muñoz, Hermógenes; Castilla-Llorente, Cristina; Plaza, Eva M; Martínez-Millán, Cristina; Heras, Inmaculada; Iniesta, Pastora; Amigo, María L; Ferrer-Marin, Francisca; Candela, María J; Lozano, María L; Vicente, Vicente; Rivera, José

    2017-04-13

    Transfusion of washed platelet concentrates (W-PC) is recommended for some patients, such as those who have had previous severe allergic transfusion reactions. However, we still lack a standardised method for preparing these products. Here, we assessed the effect of a manual washing procedure on in vitro platelet quality and on the transfusion efficacy of W-PCs. Buffy coat-derived W-PC in Composol solution were prepared by onestep centrifugation. Platelet activation and function were evaluated before and after washing by means of: (i) CD62 expression by flow cytometry; (ii) platelet aggregation (LTA); and (iii) the VerifyNow(®) P2Y12 test. A pilot prospective transfusion study was carried out in 11 onco-hematology patients receiving, in a short time, two consecutive transfusions: one with standard PC (S-PC) and one with W-PC. The post-transfusion platelet increment, the 1 h and 24 h corrected count increment (CCI) and occurrence of bleeding events were used as indices of transfusion efficacy. Platelet recovery in W-PC was 84.8±5.4%. Washing slightly increased platelet activation in W-PC vs pre-washed samples (% CD62+ platelets 23.6±7 vs 14.8±1; p=0.03). As compared to prewash samples, platelet reactivity of W-PC as measured by VerifyNow(®) P2Y12 was significantly lower with ADP (PRU 32.2±37.7 vs 4.2±2.4, p=0.027), but similar using TRAP. Platelet aggregation responses to TRAP, collagen, ristocetin and arachidonic acid were maintained in W-PC. The pilot transfusion trial showed similar 1 h (13.5±5.6 vs 11.5±7.3, p=0.49) and 24 h (11±7.2 vs 9±6.5, p=0.48) CCI for S-PC and W-PC. Transfusion of W-PC was not associated with an increased number of bleeding events. We have set up a simple method to obtain buffy-coat-derived W-PC, which has minor effects on in vitro platelet quality and transfusion effectiveness. This procedure can be easily implemented in transfusion centres for on-demand preparation of washed platelets.

  2. Evaluation of two detection methods of microorganisms in platelet concentrates.

    PubMed

    Albertoni, G; Andrade, S S; Araújo, P R B; Carvalho, F O; Girão, M J B C; Barreto, J A

    2011-12-01

    The performance of a bacterial 16S ribosomal DNA real-time polymerase chain reaction (PCR) assay was evaluated and validated with an automated culture system to determine its use for screening of platelet concentrates (PCs). PCs were spiked with suspensions of Escherichia coli, Serratia marcescens, Staphylococcus epidermidis and St. aureus at 1, 10, and 100 colony-forming units (CFUs) mL and stored for 5 days. DNA amplification was performed using real-time PCR. The BacT/ALERT was used as a reference method and samples were inoculated into an aerobic culture bottle; for the PCR assay, aliquots were drawn from all (spiked) PCs on days 0 to 5 of storage. Real-time PCR detected only the gram-positive bacteria in PCs spiked with low bacterial titres (1 CFU mL) after 48 h; however, it was able to detect all positive samples in PCs spiked with 10 CFU mL of either gram-positive or gram-negative bacteria after 48 h. In addition, real-time PCR detected all positive samples in PCs spiked with high gram-positive bacterial titres (100 CFU mL) after 24 h. On the other hand, the BacT/ALERT system showed positive results in all samples within 24 h. The BacT/ALERT method is more sensitive and should continue to be the gold standard for identifying bacterial contaminations in blood samples. The real-time PCR approach can be used for the screening of PCs for microbial detection before they are released from blood centres or shortly before they are used in blood transfusion, and thus allow an extended shelf life of the platelets. © 2011 The Authors. Transfusion Medicine © 2011 British Blood Transfusion Society.

  3. Nanomolar concentrations of adrenaline induce platelet adhesion in vitro.

    PubMed

    Eriksson, Andreas C; Whiss, Per A

    2013-01-01

    Adrenaline is a platelet activator having a resting plasma concentration of <1 nmol/l that increases to a few nmol/l during stress. However, most in vitro assays only detect effects of adrenaline in micromolar concentrations. This makes it difficult to estimate the relevance of in vitro data for the in vivo situation. The aim of this study was to investigate experimental conditions in vitro that could detect platelet effects of adrenaline in nanomolar concentrations. Platelet adhesion to albumin and collagen was evaluated with a static platelet adhesion assay. Our results show that 10 nmol/l adrenaline induced platelet adhesion to albumin in platelet-rich plasma (PRP) prepared at 140 × g, while 100 nmol/l was necessary in order to increase adhesion of platelets prepared at 220 × g. The mean platelet volume was increased after preparation at 140 × g, suggesting that large reactive platelets contributed to the increased adrenaline sensitivity. At optimal Mg(2+)-concentration, adhesion to collagen was increased by 10 nmol/l adrenaline irrespective of centrifugal force applied during PRP preparation. More specifically, we defined two populations where adhesion to collagen was increased by 10 nmol/l adrenaline either upon centrifugation at 140 × g but not 220 × g or vice versa. In some experiments, platelet adhesion to collagen was induced by 3 nmol/l adrenaline, which corresponds to concentrations achieved during stress in vivo. In summary, the static adhesion assay is able to detect platelet activating effects of adrenaline very close to physiological concentrations. This is rare for in vitro assays and motivates further research about adrenergic signalling in platelets.

  4. Relationships of the platelet aggregate ratio to serum cholesterol concentration, smoking and age

    PubMed Central

    Davis, James W.; Lewis, H. Daniel; Phillips, Phyllis E.; Davis, Rebecca F.

    1981-01-01

    The platelet aggregate ratio has been found to be decreased in some patients with vascular diseases suggesting the presence of increased circulating platelet aggregates. It has also been reported that hypercholesterolaemia is associated with an enhanced response of platelets to aggregating agents in platelet-rich plasma. The primary purpose of this investigation was to determine correlation of the platelet aggregate ratio with the serum cholesterol concentration of men with vascular diseases. For 52 men referred because of known or suspected coronary artery disease, cerebrovascular disease, or venous thromboembolism, the correlation coefficient of 0·06 suggested that the serum cholesterol concentration within the range observed (135-360 mg/dl) was not a factor influencing the platelet aggregate ratio. There was not a statistically significant difference between the mean platelet aggregate ratios or the mean serum cholesterol concentrations of the 21 non-smokers and the 31 smokers studied. A correlation coefficient of 0·03 between the platelet aggregate ratio and age of the patient suggested that the platelet aggregate ratio was independent of age in men with occlusive vascular diseases. PMID:7329876

  5. Extended Storage of Pathogen Reduced Platelet Concentrates (PRECON)

    DTIC Science & Technology

    2015-10-01

    Society of Hematology Annual Meeting; Orlando, FL December 5-8, 2015:184 INVENTIONS, PATENTS AND LICENSES: None REPORTABLE OUTCOMES: None OTHER...survivals. Data are reported as the average ±1 S.E.(1) Approximately 80% of the platelets given in the U.S. are transfused into hematology /oncology...Bailey SL, Gettinger. In Vivo Viability of Platelets Stored in Whole Blood at 4°C. Submitted abstract, American Society of Hematology Annual Meeting

  6. Platelet 5-HT concentration and comorbid depression in war veterans with and without posttraumatic stress disorder.

    PubMed

    Mück-Seler, Dorotea; Pivac, Nela; Jakovljević, Miro; Sagud, Marina; Mihaljević-Peles, Alma

    2003-07-01

    The serotonergic system is implicated in the pathophysiology of posttraumatic stress disorder (PTSD) and depression. The present study focused on platelet serotonin (5-HT) concentration and symptoms of comorbid depression in war veterans with or without PTSD. PTSD and depression were evaluated using Clinician Administered PTSD Scale, Davidson Trauma Scale, Montgomery-Asberg Depression Rating Scale and Hamilton Anxiety Scale. Sixty-five male drug-free war veterans (48 with PTSD and 17 without PTSD) and 65 age- and sex-matched healthy controls were studied. Comorbid depression occurred in 54 and 31% of war veterans with PTSD and without PTSD, respectively. Platelet 5-HT concentration was similar in the groups of depressed and nondepressed war veterans with or without PTSD and healthy controls. Platelet 5-HT concentration was found to differ between war veterans with various degrees of appetite loss. A positive correlation was observed between platelet 5-HT concentration and severity of appetite loss in veterans with PTSD. There was no relationship between platelet 5-HT concentration and severity of other symptoms of PTSD or depression. War veterans included in the study were outpatients. War veterans with PTSD had a high incidence of comorbid depression, that was not related to platelet 5-HT concentration. The marked relationship between platelet 5-HT concentration and severity of appetite loss, suggested that 5-HT system is involved in the regulation of appetite, at least in depressed war veterans with PTSD.

  7. Effect of hyperbaric oxygen therapy combined with autologous platelet concentrate applied in rabbit fibula fraction healing

    PubMed Central

    Neves, Paulo César Fagundes; de Campos Vieira Abib, Simone; Neves, Rogério Fagundes; Pircchio, Oronzo; Saad, Karen Ruggeri; Saad, Paulo Fernandes; Simões, Ricardo Santos; Moreira, Marcia Bento; de Souza Laurino, Cristiano Frota

    2013-01-01

    OBJECTIVES: The purpose is to study the effects of hyperbaric oxygen therapy and autologous platelet concentrates in healing the fibula bone of rabbits after induced fractures. METHODS: A total of 128 male New Zealand albino rabbits, between 6–8 months old, were subjected to a total osteotomy of the proximal portion of the right fibula. After surgery, the animals were divided into four groups (n = 32 each): control group, in which animals were subjected to osteotomy; autologous platelet concentrate group, in which animals were subjected to osteotomy and autologous platelet concentrate applied at the fracture site; hyperbaric oxygen group, in which animals were subjected to osteotomy and 9 consecutive daily hyperbaric oxygen therapy sessions; and autologous platelet concentrate and hyperbaric oxygen group, in which animals were subjected to osteotomy, autologous platelet concentrate applied at the fracture site, and 9 consecutive daily hyperbaric oxygen therapy sessions. Each group was divided into 4 subgroups according to a pre-determined euthanasia time points: 2, 4, 6, and 8 weeks postoperative. After euthanasia at a specific time point, the fibula containing the osseous callus was prepared histologically and stained with hematoxylin and eosin or picrosirius red. RESULTS: Autologous platelet concentrates and hyperbaric oxygen therapy, applied together or separately, increased the rate of bone healing compared with the control group. CONCLUSION: Hyperbaric oxygen therapy and autologous platelet concentrate combined increased the rate of bone healing in this experimental model. PMID:24141841

  8. A Novel Platelet Concentrate: Titanium-Prepared Platelet-Rich Fibrin

    PubMed Central

    Tunalı, Mustafa; Özdemir, Hakan; Küçükodacı, Zafer; Akman, Serhan; Yaprak, Emre; Toker, Hülya; Fıratlı, Erhan

    2014-01-01

    We developed a new product called titanium-prepared platelet-rich fibrin (T-PRF). The T-PRF method is based on the hypothesis that titanium may be more effective in activating platelets than the silica activators used with glass tubes in Chouckroun's leukocyte- and platelet-rich fibrin (L-PRF) method. In this study, we aimed to define the structural characteristics of T-PRF and compare it with L-PRF. Blood samples were collected from 10 healthy male volunteers. The blood samples were drawn using a syringe. Nine milliliters was transferred to a dry glass tube, and 9 mL was transferred to a titanium tube. Half of each clot (i.e., the blood that was clotted using T-PRF or L-PRF) was processed with a scanning electron microscope (SEM). The other half of each clot was processed for fluorescence microscopy analysis and light microscopy analysis. The T-PRF samples seemed to have a highly organized network with continuous integrity compared to the other L-PRF samples. Histomorphometric analysis showed that T-PRF fibrin network covers larger area than L-PRF fibrin network; also fibrin seemed thicker in the T-PRF samples. This is the first human study to define T-PRF as an autogenous leukocyte- and platelet-rich fibrin product. The platelet activation by titanium seems to offer some high characteristics to T-PRF. PMID:24563860

  9. Use of platelet concentrates in oral and maxillofacial surgery: an overview.

    PubMed

    Mihaylova, Zornitsa; Mitev, Vanyo; Stanimirov, Pavel; Isaeva, Antonia; Gateva, Natalia; Ishkitiev, Nikolay

    2017-01-01

    To describe and provide a comprehensive overview on the development, use and efficacy of autologous platelet concentrates in different in vitro and in vivo studies focusing on oral and maxillofacial pathologies. Present work employs an extensive critical overview of the literature on the development and application of platelet concentrates. Platelet concentrates are innovative endogenous therapeutic agents which gained a lot of interest in different medical and dental disciplines due to their potential ability to stimulate and increase regeneration of soft and hard tissues. The effect of platelet-derived products is considered to be a result of the high number of platelets which contain a wide range of growth factors. They are not just therapeutic products but autologous blood concentrates containing active molecules. The quality of platelet concentrates may vary according to the individual physical state of donors making it difficult to to compare the outcomes of their application. Although, there are many studies analyzing the properties of these biomaterials both in vivo and in vitro, a consensus regarding their efficacy still has to be reached. Evidences described in the literature on the efficacy of platelet concentrates in procedures in oral and maxillofacial region are controversial and limited. In order to clarify the real advantages and priorities for the patients, when the blood-derived products are applied, further in vitro and in vivo research about the activity of PRP and PRF on the dental cells biology should be conducted.

  10. Platelet-rich plasma releasate differently stimulates cellular commitment toward the chondrogenic lineage according to concentration

    PubMed Central

    Matsiko, Amos; Tomazette, Marcel RP; Rocha, Wanessa KR; Cordeiro-Spinetti, Eric; Levingstone, Tanya J; Farina, Marcos; O’Brien, Fergal J; El-Cheikh, Marcia C; Balduino, Alex

    2015-01-01

    Platelet-rich plasma has been used to treat articular cartilage defects, with the expectations of anabolic and anti-inflammatory effects. However, its role on cellular chondrogenic or fibrogenic commitment is still a controversy. Herein, the role of platelet-rich plasma releasate, the product obtained following platelet-rich plasma activation, on cellular commitment toward the chondrogenic lineage was evaluated in vitro. Human nasoseptal chondrogenic cells and human bone marrow mesenchymal stromal cells were used as cell types already committed to the chondrogenic lineage and undifferentiated cells, respectively, as different concentrations of platelet-rich plasma releasate were tested in comparison to commonly used fetal bovine serum. Low concentration of platelet-rich plasma releasate (2.5%) presented similar effects on cellular growth compared to 10% fetal bovine serum, for both cell types. In a three-dimensional culture system, platelet-rich plasma releasate alone did not induce full nasoseptal chondrogenic cells cartilage-like pellet formation. Nonetheless, platelet-rich plasma releasate played a significant role on cell commitment as high-passage nasoseptal chondrogenic cells only originated cartilage-like pellets when expanded in the presence of platelet-rich plasma releasate rather than fetal bovine serum. Histological analyses and measurements of pellet area demonstrated that even low concentrations of platelet-rich plasma releasate were enough to prevent nasoseptal chondrogenic cells from losing their chondrogenic potential due to in vitro expansion thereby promoting their recommitment. Low concentration of platelet-rich plasma releasate supplemented in chondrogenic medium also increased the chondrogenic potential of mesenchymal stromal cells seeded on collagen-hyaluronic acid scaffolds, as observed by an increase in chondrogenic-related gene expression, sulfated glycosaminoglycan production, and compressive modulus following in vitro culture. On the

  11. Effects of plasma nitric oxide levels on platelet activation in single donor apheresis and random donor concentrates.

    PubMed

    Büyükkağnici, Demet Iren; Ilhan, Osman; Kavas, Güzin Ozelçi; Arslan, Onder; Arat, Mutlu; Dalva, Klara; Ayyildiz, Erol

    2007-02-01

    P-selectin is an useful marker to determine platelet activation and nitric oxide inhibits platelet activation, secretion, adhesion and aggregation. The aim of this study was to investigate the relationship between nitric oxide and P-selectin values in both single donor apheresis and random donor platelet concentrates. According to the results of this study, we found that the best platelet concentrate is freshly prepared single donor apheresis concentrate and it is important to prevent activation at the beginning of the donation. Nitric oxide, which is synthesized from platelets during the storage period, is not sufficient to prevent platelet activation.

  12. Surface modification of platelet concentrate bags to reduce biofilm formation and transfusion sepsis.

    PubMed

    Wilson-Nieuwenhuis, Joels S T; Dempsey-Hibbert, Nina; Liauw, Christopher M; Whitehead, Kathryn A

    2017-09-07

    Bacterial contamination of blood products poses a major risk in transfusion medicine, including transfusions involving platelet products. Although testing systems are in place for routine screening of platelet units, the formation of bacterial biofilms in such units may decrease the likelihood that bacteria will be detected. This work determined the surface properties of p-PVC platelet concentrate bags and investigated how these characteristics influenced biofilm formation. Serratia marcescens and Staphylococcus epidermidis, two species commonly implicated in platelet contamination, were used to study biofilm growth. The platelet concentrate bags were physically flattened to determine if reducing the surface roughness altered biofilm formation. The results demonstrated that the flattening process of the platelet bags affected the chemistry of the surface and reduced the surface hydrophobicity. Flattening of the surfaces resulted in a reduction in biofilm formation for both species after 5 days, with S. marcescens demonstrating a greater reduction. However, there was no significant difference between the smooth and flat surfaces following 7 days' incubation for S. marcescens and no significant differences between any of the surfaces following 7 days' incubation for S. epidermidis. The results suggest that flattening the p-PVC surfaces may limit potential biofilm formation for the current duration of platelet storage time of 5 days. It is hoped that this work will enhance the understanding of how surface properties influence the development of microbial biofilms in platelet concentrate bags in order to devise a solution to discourage biofilm formation. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Diabetic concentrations of metformin inhibit platelet-mediated ovarian cancer cell progression.

    PubMed

    Erices, Rafaela; Cubillos, Sofía; Aravena, Raúl; Santoro, Felice; Marquez, Monica; Orellana, Renan; Ramírez, Carolina; González, Pamela; Fuenzalida, Patricia; Bravo, María Loreto; Oliva, Bárbara; Kato, Sumie; Ibañez, Carolina; Brañes, Jorge; Bravo, Erasmo; Alonso, Catalina; García, Karen; Arab, Clemente; Torres, Vicente A; Godoy, Alejandro S; Pereira, Jaime; Bustos, Galdo; Cardenas, Julio Cesar; Cuello, Mauricio A; Owen, Gareth I

    2017-02-15

    Clinical studies have suggested a survival benefit in ovarian cancer patients with type 2 diabetes mellitus taking metformin, however the mechanism by which diabetic concentrations of metformin could deliver this effect is still poorly understood. Platelets not only represent an important reservoir of growth factors and angiogenic regulators, they are also known to participate in the tumor microenvironment implicated in tumor growth and dissemination. Herein, we investigated if diabetic concentrations of metformin could impinge upon the previously reported observation that platelet induces an increase in the tube forming capacity of endothelial cells (angiogenesis) and upon ovarian cancer cell aggressiveness. We demonstrate that metformin inhibits the increase in angiogenesis brought about by platelets in a mechanism that did not alter endothelial cell migration. In ovarian cancer cell lines and primary cultured cancer cells isolated from the ascitic fluid of ovarian cancer patients, we assessed the effect of combinations of platelets and metformin upon angiogenesis, migration, invasion and cancer sphere formation. The enhancement of each of these parameters by platelets was abrogated by the present of metformin in the vast majority of cancer cell cultures tested. Neither metformin nor platelets altered proliferation; however, metformin inhibited the increase in phosphorylation of focal adhesion kinase induced by platelets. We present the first evidence suggesting that concentrations of metformin present in diabetic patients may reduce the actions of platelets upon both endothelial cells and cancer cell survival and dissemination.

  14. Diabetic concentrations of metformin inhibit platelet-mediated ovarian cancer cell progression

    PubMed Central

    Erices, Rafaela; Cubillos, Sofía; Aravena, Raúl; Santoro, Felice; Marquez, Monica; Orellana, Renan; Ramírez, Carolina; González, Pamela; Fuenzalida, Patricia; Bravo, María Loreto; Oliva, Bárbara; Kato, Sumie; Ibañez, Carolina; Brañes, Jorge; Bravo, Erasmo; Alonso, Catalina; García, Karen; Arab, Clemente; Torres, Vicente A.; Godoy, Alejandro S.; Pereira, Jaime; Bustos, Galdo; Cardenas, Julio Cesar; Cuello, Mauricio A.; Owen, Gareth I.

    2017-01-01

    Clinical studies have suggested a survival benefit in ovarian cancer patients with type 2 diabetes mellitus taking metformin, however the mechanism by which diabetic concentrations of metformin could deliver this effect is still poorly understood. Platelets not only represent an important reservoir of growth factors and angiogenic regulators, they are also known to participate in the tumor microenvironment implicated in tumor growth and dissemination. Herein, we investigated if diabetic concentrations of metformin could impinge upon the previously reported observation that platelet induces an increase in the tube forming capacity of endothelial cells (angiogenesis) and upon ovarian cancer cell aggressiveness. We demonstrate that metformin inhibits the increase in angiogenesis brought about by platelets in a mechanism that did not alter endothelial cell migration. In ovarian cancer cell lines and primary cultured cancer cells isolated from the ascitic fluid of ovarian cancer patients, we assessed the effect of combinations of platelets and metformin upon angiogenesis, migration, invasion and cancer sphere formation. The enhancement of each of these parameters by platelets was abrogated by the present of metformin in the vast majority of cancer cell cultures tested. Neither metformin nor platelets altered proliferation; however, metformin inhibited the increase in phosphorylation of focal adhesion kinase induced by platelets. We present the first evidence suggesting that concentrations of metformin present in diabetic patients may reduce the actions of platelets upon both endothelial cells and cancer cell survival and dissemination. PMID:28209916

  15. Possible implication of sterile connecting device in contamination of pooled platelet concentrates.

    PubMed

    Mertens, G; Muylle, L; Goossens, H

    1997-09-01

    Considering the possibility that a pooled random donor platelet concentrate could become contaminated by welding with a sterile connecting device, we undertook a study to determine the influence of pooling on the contamination rate. As a control group, apheresis platelets were examined. Bacteriological testing was done with a sensitive CO2 detecting culture system, the BacT/ Alert. Out of 1105 pooled platelet concentrates prepared by the buffy coat method, 15 (1.4%) were confirmed as contaminated, all with Staphylococcus epidermidis and two with a second bacterial species, i.e. Staphylococcus capitis and Propionibacterium acnes, respectively. Median detection time by the BacT/Alert was 23 h. Twelve pools of five units were contaminated, which is significantly more than the three contaminated pools of four units. On the other hand, the reuse of the welding wafers proved not be a risk factor for contamination. One welded tubing segment of a contaminated platelet concentrate failed the air leakage test, an incident which was 73 times more frequent than with the sterile platelet concentrates. We found five pooled platelet concentrates containing Staphylococci from which no bacteria could be grown from the individual buffy coats that had been pooled. We suggest the contamination here to have occurred after separation of the buffy coat from the whole blood, possibly during the welding process. Finally, none out of 378 apheresis platelet concentrates was contaminated. All our observations highlight the potential risk for contamination when making pooled platelet concentrates with a sterile connecting device. For this type of transfusion product, we advocate bacteriological screening of all units before release. The incubation time for the sterility test should, however, be limited to 36 h, if logistical problems with the availability of platelets are to be avoided.

  16. Platelet Activation by Low Concentrations of Intact Oxidized LDL Particles Involves the PAF Receptor

    PubMed Central

    Chen, Rui; Chen, Xi; Salomon, Robert G.; McIntyre, Thomas M.

    2008-01-01

    Objective Mitochondrial depolarization aids platelet activation. Oxidized LDL (oxLDL) contains the medium length oxidatively truncated phospholipid hexadecyl azelaoyl-lysoPAF (HAz-LPAF) that disrupts mitochondrial function in nucleated cells, so oxLDL may augment platelet activation. Methods and Results Flow cytometry showed intact oxLDL particles synergized with sub-threshold amounts of soluble agonists to increase intracellular Ca++, and initiate platelet aggregation and surface expression of activated gpIIb/IIIa and P-selectin. oxLDL also induced aggregation and increased intracellular Ca++ in FURA2-labeled cells by itself at low, although not higher, concentrations. HAz-LPAF, alone and in combination with sub-stimulatory amounts of thrombin, rapidly increased cytoplasmic Ca++ and initiated aggregation. HAz-LPAF depolarized mitochondria in intact platelets, but this required concentrations beyond those that directly activated platelets. An unexpectedly large series of chemically pure truncated phospholipids generated by oxidative fragmentation of arachidonoyl-, docosahexaneoyl-, or linoleoyl alkyl phospholipids were platelet agonists. The PAF receptor, thought to effectively recognize only phospholipids with very short sn-2 residues, was essential for platelet activation because PAF receptor agonists blocked signaling by all these medium length phospholipids and oxLDL. Conclusions Intact oxLDL particles activate platelets through the PAF receptor, and the PAF receptor responds to a far wider range of oxidized phospholipids in oxLDL than anticipated. PMID:19112165

  17. Platelet activating factor raises intracellular calcium ion concentration in macrophages

    PubMed Central

    1986-01-01

    Peritoneal cells from thioglycollate-stimulated mice were allowed to adhere to coverglasses for 2 h to give a dense monolayer of adherent cells greater than 95% of which were macrophages. After incubation with the tetra-acetoxymethyl ester of quin2, coverglasses were rinsed with Ca2+-free saline, oriented at a 45 degree angle in square cuvettes containing a magnetically driven stir bar, and analyzed for changes in quin2 fluorescence in a spectrofluorimeter. Such fluorescence, taken as an indication of intracellular calcium ion concentration ([Ca2+]i), increased as exogenous calcium ion concentration ([Ca2+]o) was raised to 1 mM. At [Ca2+]o approximately equal to 10 microM, [Ca2+]i = 72 +/- 14 nM (n = 26); at [Ca2+]o = 1 mM, [Ca2+]i = 140-220 nM, levels not increased by N, N, N', N'-tetrakis (2-pyridylmethyl) ethylenediamine, a membrane-permeant chelator of heavy metals than can quench quin2. Addition of mouse alpha + beta fibroblast interferon, lipopolysaccharide, thrombin, collagen, vasopressin, ADP, compound 48/80, or U46619 did not change [Ca2+]i. However, addition of platelet activating factor (PAF) (2-20 ng/ml) raised [Ca2+]i by 480 nM within 1 min if [Ca2+]o = 1 mM. In the presence of 5 mM EGTA, PAF raised [Ca2+]i by 25 nM. This suggests that PAF causes influx of exogenous Ca2+, as well as releasing some Ca2+ from intracellular stores. Consistent with these results, when PAF was added to 1 mM Ca2+ in the presence of 100 microM Cd2+ or Mn2+ to block Ca2+ influx, [Ca2+]i increased by only intermediate amounts; at the times of such dampened peak response, [Ca2+]i could be raised within 1 min to normal PAF-stimulated levels by chelation of the exogenous heavy metals with diethylenetriaminepentaacetic acid. Normal PAF responses were observed in the presence of indomethacin. The lowest dose of PAF observed to raise [Ca2+]i was 0.1 ng/ml. Response of [Ca2+]i to 2-20 ng/ml PAF was transient, and second applications had no effect. The PAF response also was seen in

  18. 5-day storage of platelet concentrates in CLX containers: effect of type of agitation.

    PubMed

    Snyder, E L; Bookbinder, M; Kakaiya, R; Ferri, P; Kiraly, T

    1983-01-01

    To determine the degree of platelet damage produced by different modes of agitation during storage of concentrates for 5 days in CLX blood bags, we studied pH, platelet counts, release of LDH and beta thromboglobulin, morphology and osmotic recovery. Platelets were maintained at 20-24 degrees C on elliptical, 6-rpm circular, 2-rpm circular and flat bed agitators. At 72-120 h platelet concentrates stored on the flat bed shaker had significantly lower pH values than units stored on the elliptical or on either of the circular rotators (p less than 0.05). The percent LDH discharged was highest for the units stored on the elliptical rotator (p less than 0.05). Remaining tests of platelet function were not significantly different for concentrates stored on any of the four agitators. Flat bed shakers were unable to resuspend the platelet 'button' which formed after the final preparative centrifugation. Based on our in vitro studies, we conclude that due to problems with low pH values, flat bed shakers may not be optimal for storing platelet concentrates in CLX blood bags and that some other form of agitation should be used.

  19. Effects of different aspirin formulations on platelet aggregation times and on plasma salicylate concentrations.

    PubMed

    Schwertner, H A; McGlasson, D; Christopher, M; Bush, A C

    2006-01-01

    Early aspirin treatment is widely used to inhibit platelet activity and to reduce morbidity and mortality in patients presenting with an acute myocardial infarction or a stroke. A number of different aspirin formulations have been used for this purpose; however, a comparison of their effectiveness in inhibiting early platelet aggregation has not been determined. In this study, we determined plasma salicylate concentrations and platelet inhibitory activities at various times after ingestion of three commonly used aspirin formulations: soluble aspirin (Alka-Seltzer), 325 mg, chewed baby aspirin, 324 mg, and whole compressed non-enteric coated aspirin, 324 mg. Twenty-four healthy volunteers, 18-39 years of age, participated in the prospective single-blinded triple-crossover study. Plasma salicylate concentrations and inhibition of arachidonic acid-induced platelet aggregation were determined on post-dose blood samples collected at 2.5, 5.0, 7.5, 10, 15, 20, 25, 30, and 40 min. All subjects crossed over to the other two formulations with at least 2 weeks between ingestions. The median platelet inhibition times for the chewed, soluble, and whole aspirin formulations were 7.5, 7.5, and 10.0 min, respectively. Soluble and chewed aspirin were found to inhibit platelet aggregation faster than whole aspirin (p<0.001); however, there were no significant differences in platelet aggregation times between the soluble and chewed formulations (p<0.163). Inhibition of platelet aggregation was found to occur at an average plasma salicylate concentration of 2.46 microg/mL, regardless of method of ingestion. The results indicate that soluble and chewed aspirin inhibit platelet aggregation in a shorter period of time than does whole aspirin. The results suggest that chewing baby aspirin or taking soluble buffered aspirin may be the preferred route of administration for early platelet inhibition.

  20. Platelet serotonin concentration and suicidal behavior in combat related posttraumatic stress disorder.

    PubMed

    Kovacic, Zrnka; Henigsberg, Neven; Pivac, Nela; Nedic, Gordana; Borovecki, Andrea

    2008-02-15

    Posttraumatic stress disorder (PTSD) is a serious and global problem, a psychiatric disorder that frequently occurs with different comorbidities, and is associated with a high suicide rate. Pathophysiologically, both PTSD and suicidal behavior are related to disturbances in the central serotonergic system. Serotonin (5-hydroxytryptamine, 5-HT) controls emotional behavior, anxiety, impulsivity and aggression, and nearly all known antidepressants and antianxiety drugs affect 5-HT transmission. Platelet 5-HT can be used as a limited peripheral marker of the central serotonergic synaptosomes, since it is related to particular basic psychopathological characteristics of several psychiatric disorders. Platelet 5-HT concentration has been reported to be similar in PTSD subjects and healthy controls, but suicidal patients across different psychiatric diagnoses have reduced platelet 5-HT concentration. This study examined platelet 5-HT concentration by the spectrofluorimetric method in male subjects: 73 suicidal and 47 non-suicidal veterans with current and chronic combat related PTSD, 45 suicidal and 30 non-suicidal comparative non-PTSD subjects and 147 healthy men. The presence of suicidal behavior (score=0, non-suicidal; scores > or =1, suicidal) was assessed with the Hamilton Depression Rating Scale-17 (HDRS). Platelet 5-HT concentration was significantly lower in suicidal PTSD and non-PTSD patients compared to non-suicidal patients or healthy controls. Since the majority of patients scored very low on item 3 of HDRS, no significant correlation between suicidal scores and platelet 5-HT concentration was found. These results show that reduced platelet 5-HT concentration is related to suicidal behavior in PTSD, and suggest that platelet 5-HT concentration might be used as a peripheral marker to predict suicidal behavior across psychiatric diagnoses.

  1. Determination of thromboxane formation, soluble CD40L release and thrombopoietin clearance in apheresis platelet concentrates.

    PubMed

    Wenzel, Folker; Baertl, Anja; Hohlfeld, Thomas; Zimmermann, Norbert; Weber, Artur Aron; Lorenz, Horst; Giers, Günther

    2012-01-01

    All deleterious changes in platelet morphology, structure and function that occur in platelet concentrates (PC) during storage are titled as the 'platelet storage lesion'. No single in vitro test currently available is sufficient in assessing these changes of platelet quality. The release of soluble CD40 Ligand (sCD40L), the formation of thromboxane (TXB2) and the thrombopoietin (TPO) clearance reflect different aspects of platelet metabolism and activitiy, and were used to examine platelet quality in apheresis platelet products. At days 1, 3 and 5, in single-donor apheresis platelet products (n = 10) under routine storage conditions, sCD40L (measured by ELISA) and TXB2 (measured by RIA) were determined after platelet stimulation (recalcification and clot formation). TPO (measured by ELISA) was determined after an incubation time of 5 h at 37°C with platelet-rich plasma (adjusted initial TPO concentration of about 500 pg/mL). Results were related to a therapeutic unit (TU = 2 × 10(11) platelets). Immediately after platelet preparation, sCD40L release was 41 ± 7.6 ng/TU, TXB2 formation 1688 ± 374 ng/TU and TPO clearance 1.22 ± 0.32 ng/h/TU. At days 1, 3 and 5, sCD40L was reduced to 89 ± 7%, 71 ± 12% and 57 ± 9%, TXB2 release to 91 ± 6%, 74 ± 12% and 58 ± 9% and TPO clearance to 90 ± 15%, 84 ± 5% and 79 ± 10% of the respective control values. In conclusion, in single-donor apheresis PC, sCD40L release and TXB2 formation as well as TPO clearance by the platelets were dependent on storage duration and reduced to about 60% to 80% of the respective control values after a storage period for 5 days. These findings are in line with literature data, indicating that a loss of platelet functionality of about 30% will occur after 5 days of storage.

  2. Effects of anticoagulant on pH, ionized calcium concentration, and agonist-induced platelet aggregation in canine platelet-rich plasma

    PubMed Central

    Callan, Mary Beth; Shofer, Frances S.; Catalfamo, James L.

    2014-01-01

    Objective To compare effects of 3.8% sodium citrate and anticoagulant citrate dextrose solution National Institutes of Health formula A (ACD-A) on pH, extracellular ionized calcium (iCa) concentration, and platelet aggregation in canine platelet-rich plasma (PRP). Sample Population Samples from 12 dogs Procedures Blood samples were collected into 3.8% sodium citrate (dilution, 1:9) and ACD-A (dilution, 1:5). Platelet function, pH, and iCa concentration were evaluated in PRP Platelet agonists were ADP γ-thrombin, and convulxin; final concentrations of each were 20μM, 100nM, and 20nM, respectively. Washed platelets were used to evaluate effects of varying the pH and iCa concentration. Results Mean pH and iCa concentration were significantly greater in 3.8% sodium citrate PRP than ACD-A PRP Platelet aggregation induced by ADP and γ-thrombin was markedly diminished in ACD-A PRP compared with results for 3.8% sodium citrate PRP Anticoagulant had no effect on amplitude of convulxin-induced platelet aggregation. In washed platelet suspensions (pH, 7.4), there were no differences in amplitude of platelet aggregation induced by convulxin or γ-thrombin at various iCa concentrations. Varying the pH had no effect on amplitude of aggregation induced by convulxin or γ-thrombin, but the aggregation rate increased with increasing pH for both agonists. Conclusions and Clinical Relevance Aggregation of canine platelets induced by ADP and γ-thrombin was negligible in ACD-A PRP which suggested an increase in extraplatelet hydrogen ion concentration inhibits signaling triggered by these agonists but not by convulxin. Choice of anticoagulant may influence results of in vitro evaluation of platelet function, which can lead to erroneous conclusions. PMID:19335102

  3. Effects of anticoagulant on pH, ionized calcium concentration, and agonist-induced platelet aggregation in canine platelet-rich plasma.

    PubMed

    Callan, Mary Beth; Shofer, Frances S; Catalfamo, James L

    2009-04-01

    OBJECTIVE-To compare effects of 3.8% sodium citrate and anticoagulant citrate dextrose solution National Institutes of Health formula A (ACD-A) on pH, extracellular ionized calcium (iCa) concentration, and platelet aggregation in canine platelet-rich plasma (PRP). SAMPLE POPULATION-Samples from 12 dogs. PROCEDURES-Blood samples were collected into 3.8% sodium citrate (dilution, 1:9) and ACD-A (dilution, 1:5). Platelet function, pH, and iCa concentration were evaluated in PRP. Platelet agonists were ADP, gamma-thrombin, and convulxin; final concentrations of each were 20microm, 100nM, and 20nM, respectively. Washed platelets were used to evaluate effects of varying the pH and iCa concentration. RESULTS-Mean pH and iCa concentration were significantly greater in 3.8% sodium citrate PRP than ACD-A PRP. Platelet aggregation induced by ADP and gamma-thrombin was markedly diminished in ACD-A PRP, compared with results for 3.8% sodium citrate PRP. Anticoagulant had no effect on amplitude of convulxin-induced platelet aggregation. In washed platelet suspensions (pH, 7.4), there were no differences in amplitude of platelet aggregation induced by convulxin or gamma-thrombin at various iCa concentrations. Varying the pH had no effect on amplitude of aggregation induced by convulxin or gamma-thrombin, but the aggregation rate increased with increasing pH for both agonists. CONCLUSIONS AND CLINICAL RELEVANCE-Aggregation of canine platelets induced by ADP and gamma-thrombin was negligible in ACD-A PRP, which suggested an increase in extraplatelet hydrogen ion concentration inhibits signaling triggered by these agonists but not by convulxin. Choice of anticoagulant may influence results of in vitro evaluation of platelet function, which can lead to erroneous conclusions.

  4. Feasibility and Efficiency of Human Bone Marrow Stromal Cell Culture with Allogeneic Platelet Lysate-Supplementation for Cell Therapy against Stroke

    PubMed Central

    Tan, Chengbo; Wang, Zifeng; Hamauchi, Shuji; Abumiya, Takeo; Kazumata, Ken; Ito, Tsuneo; Kudo, Kohsuke; Takamoto, Shigeru; Houkin, Kiyohiro

    2016-01-01

    Currently, there is increasing interest in human bone marrow stromal cells (hBMSCs) as regeneration therapy against cerebral stroke. The aim of the present study was to evaluate the feasibility and validity of hBMSC cultures with allogeneic platelet lysates (PLs). Platelet concentrates (PC) were harvested from healthy volunteers and made into single donor-derived PL (sPL). The PL mixtures (mPL) were made from three different sPL. Some growth factors and platelet cell surface antigens were detected by enzyme-linked immunosorbent assay (ELISA). The hBMSCs cultured with 10% PL were analyzed for their proliferative potential, surface markers, and karyotypes. The cells were incubated with superparamagnetic iron oxide (SPIO) agents and injected into a pig brain. MRI and histological analysis were performed. Consequently, nine lots of sPL and three mPL were prepared. ELISA analysis showed that PL contained adequate growth factors and a particle of platelet surface antigens. Cell proliferation capacity of PLs was equivalent to or higher than that of fetal calf serum (FCS). No contradiction in cell surface markers and no chromosomal aberrations were found. The MRI detected the distribution of SPIO-labeled hBMSCs in the pig brain. In summary, the hBMSCs cultured with allogeneic PL are suitable for cell therapy against stroke. PMID:27840648

  5. The influence of epidural analgesia on platelet function and correlation with plasma bupivacaine concentrations.

    PubMed

    Odoom, J A; Dokter, P W; Sturk, A; Ten Cate, J W; Sih, I L; Bovill, J G

    1988-09-01

    The effect of epidural anaesthesia with bupivacaine 0.5% on platelet aggregation was studied in seven patients undergoing transurethral resection of the prostate. Peak plasma concentrations of bupivacaine 470 +/- 270 ng ml-1 occurred at 30 min after administration. At that time there were no significant changes in platelet aggregation. However, the maximum rate of the primary- and secondary-aggregation velocities induced by 1.0 microM ADP were significantly decreased at 1 h and 3 h after bupivacaine administration. The maximum percentage ADP-induced platelet aggregation was also decreased significantly at 1 h and 3 h. The minimum concentration of ADP required to induce secondary-phase platelet aggregation was significantly increased at 1 h but not at 3 h. There was a significant correlation between bupivacaine concentrations and all platelet aggregation parameters except the maximum ADP-induced aggregation. Platelet inhibition occurred at plasma bupivacaine concentrations that were considerably lower than those needed to produce similar inhibition in vitro.

  6. Application of a real-time biosensor to detect bacteria in platelet concentrates.

    PubMed

    Rotman, Boris; Cote, Mindy A

    2003-01-03

    A spore-based biosensor for detecting low levels of bacteria in real-time has been recently developed. The system (termed LEXSAS, label-free exponential signal-amplification system) exploits spore's ability to produce fluorescence when sensing neighboring bacterial cells. We studied the LEXSAS as a possible approach for identifying bacterially contaminated platelet concentrates prior to transfusion because the system offers rapid analysis, high sensitivity, and low cost. If successful, this approach could reduce the risk of morbidity and mortality from transfusion-related bacteremia and sepsis. In this study, we used the LEXSAS for detecting bacteria in platelet concentrates spiked with Bacillus cereus, Enterobacter cloacae, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, or Streptococcus pyogenes. Bacteria were separated from platelets using a 2-min procedure based on bacterial resistance to detergents and osmotic shock. The results indicate that the LEXSAS could be used to design a practical biosensor for identifying bacterially contaminated platelets in real-time.

  7. Microparticle and mitochondrial release during extended storage of different types of platelet concentrates.

    PubMed

    Marcoux, Geneviève; Duchez, Anne-Claire; Rousseau, Matthieu; Lévesque, Tania; Boudreau, Luc H; Thibault, Louis; Boilard, Eric

    2016-09-29

    On activation, platelets release vesicles called microparticles (MPs). MPs are heterogeneous with regard to the presence or absence of mitochondria. We quantified MPs in platelet concentrates (PCs) taking their mitochondrial content into account. Platelet-rich plasma (PRP), buffy coat (BC) and apheresis (AP) PCs were tested through 7 days of storage. A combination of flow cytometry and spanning-tree progression analysis of density-normalized events (SPADE) was used to determine MP and mitochondrial release during storage. All the PC biochemical parameters complied with transfusion standards at all times. Platelet activation markers increased during storage and were higher for PRP than other types of PCs. Concentrations of MPs and extracellular mitochondria interpreted by SPADE algorithm were significantly higher in PRP than other in PCs and were stable throughout storage. The mode of preparation, rather than storage duration, impacts the release of MPs and mitochondria in PCs.

  8. Platelet-derived Factor Concentrates with Hyaluronic Acid Scaffolds for Treatment of Deep Burn Wounds

    PubMed Central

    Minabe, Toshiharu; Yamakawa, Tomomi; Araki, Jun; Sano, Hitomi; Yoshimura, Kotaro

    2016-01-01

    Summary: A deep burn wound is a critical condition that generally necessitates vascularized tissue coverage. We performed the injection of platelet-derived factor concentrates combined with non–cross-linked hyaluronic acid scaffolds for 2 patients with critical burn wounds with bone and tendon exposure and achieved successful healing. Hyaluronic acid was considered to have served as a controlled-release carrier of platelet-derived factors, being clinically effective for the treatment of deep burn wounds. PMID:27826482

  9. Increased Platelet Concentration does not Improve Functional Graft Healing in Bio-Enhanced ACL Reconstruction

    PubMed Central

    Fleming, Braden C.; Proffen, Benedikt L.; Vavken, Patrick; Shalvoy, Matthew R.; Machan, Jason T.; Murray, Martha M.

    2014-01-01

    Purpose The use of an extra-cellular matrix scaffold (ECM) combined with platelets to enhance healing of an ACL graft (“bio-enhanced ACL reconstruction”) has shown promise in animal models. However, the effects of platelet concentration on graft healing remains unknown. The objectives of this study were to determine if increasing the platelet concentration in the ECM scaffold would; 1) improve the graft biomechanical properties, and 2) decrease cartilage damage after surgery. Methods Fifty-five adolescent minipigs were randomized to 5 treatment groups; untreated ACL transection (n=10), conventional ACL reconstruction (n=15), and bio-enhanced ACL reconstruction using 1X (n=10), 3X (n=10) or 5X (n=10) platelet-rich plasma. The graft biomechanical properties, anteroposterior (AP) knee laxity, graft histology and macroscopic cartilage integrity were measured at 15 weeks. Results The mean linear stiffness of the bio-enhanced ACL reconstruction procedure using the 1X preparation was significantly greater than traditional reconstruction while the 3X and 5X preparations were not. The failure loads of all the ACL reconstructed groups were equivalent but significantly greater than untreated ACL transection. There were no significant differences in the ligament maturity index or AP laxity between reconstructed knees. Macroscopic cartilage damage was relatively minor, though significantly less when the ECM-platelet composite was used. Conclusions Only the 1X platelet concentration improved healing over traditional ACL reconstruction. Increasing the platelet concentration from 1X to 5X in the ECM scaffold did not further improve the graft mechanical properties. The use of an ECM-platelet composite decreased the amount of cartilage damage seen after ACL surgery. PMID:24633008

  10. Use of hollow fiber membrane filtration for the removal of DMSO from platelet concentrates.

    PubMed

    Arnaud, F; Kapnik, E; Meryman, H T

    2003-05-01

    It has been hypothesized that, in addition to freezing injury, some damage to platelets may result from the cell packing that occurs during removal of the cryoprotectant. This study examined DMSO removal by fluid exchange across hollow-fiber (HF) filters as an alternative to centrifugation. The DMSO solution with or without cell suspension was passed once through the filter. The optimum exchange during unloading of DMSO was determined by varying the flow rates in the external and internal compartments of the HF filter. Initially, buffered solutions of a 5% DMSO solution in the absence of platelets were pumped into the fibers and exchanged against PBS. The residual DMSO was determined by osmometry. The exchange of DMSO across the membrane was flow dependent and also influenced by the chemical nature of the HF fibers. No protocol using a reasonable rate flow through the fibers removed more than 95% of the DMSO in a single pass. The optimum protocol was achieved with polysynthane fibers with an internal flow rate of approximately 20 mi/min and an external flow rate of 100 ml/min. Subsequently, frozen/thawed platelet concentrates in DMSO were washed using centrifugation and compared to the HF filtration method. Platelet quality was assayed by flow cytometry, cell count, morphology and osmotic stress test. Both filtration and centrifugal washing techniques resulted in comparable morphological scores and numbers of discoid cells. When agents reducing platelet activation were added, platelet quality was improved after washing by either technique. The lower platelet osmotic response with HF filtration than with centrifugation while using activation inhibitors was attributed to the remaining amount of the inhibitors. All other parameters tested were similar. The expression of CD62P was equivalent with both techniques, and centrifugation did not activate platelets more than filtration contrary to what was originally anticipated. In conclusion, platelet quality was comparable

  11. Comparison of the platelet-rich plasma and buffy coat protocols for preparation of canine platelet concentrates.

    PubMed

    Hoareau, Guillaume L; Jandrey, Karl E; Burges, Julie; Bremer, Daphne; Tablin, Fern

    2014-12-01

    Platelet (PLT) concentrates (PC) can be produced via the buffy coat (BC) or platelet-rich plasma (PRP) protocols. The 2 methods have not been compared with canine blood. The aims of the study were to compare the PLT, WBC, and RBC concentrations, in vitro PLT function, and markers of platelet storage lesion (PSL) in canine PC generated by 2 different protocols, and determine microbial growth throughout storage. PC from 8 healthy donor dogs were produced using 2 standard protocols, PRP and BC. PLT, WBC, and RBC counts, optical aggregometry assays, and PSL markers (pH, pCO2 , HCO3 , lactate and glucose concentrations, and LDH activity) were determined on storage days 0, 1, 3, 5, and 7. Aerobic and anaerobic bacterial cultures were also performed. Mean PLT counts were comparable between protocols and remained stable throughout storage up to day 7, while median WBC and RBC counts on day 0 were significantly higher in the BC-PC group (17,800 WBCs/μL; 195,000 RBCs/μL) than in the PRP-PC group (200 WBCs/μL; 10,000 RBCs/μL) (P = .012). In PRP-PC aggregometry, the median slope and amplitude in response to γ-thrombin and convulxin (+ ADP) were significantly decreased, and virtually absent in BC-PC during storage. PSL markers (lactate, LDH activity) were higher in BC-PC. Aerobic bacterial growth was observed in 2 PRP-PC and 1 BC-PC. This in vitro study suggests that PRP-PC had lesser WBC and RBC contamination and superior PLT function compared with BC-PC. In vivo studies are required to address safety and efficacy of PRP-PC. © 2014 American Society for Veterinary Clinical Pathology.

  12. Assessment of the Clinical Performance of Platelet Concentrates Treated by Pathogen Reduction Technology in Santiago de Compostela

    PubMed Central

    Vilariño, M. Dolores; Castrillo, Azucena; Campos, Alfredo; Kilian, Rachel; Villamayor, Mercedes; Cardoso, Marcia

    2017-01-01

    Introduction This study assessed the feasibility, performance, and safety of Mirasol®-treated platelet concentrates (M-PC) stored for up to 7 days. Methods This prospective observational study was approved by the ethical committee of the University Clinic of Santiago de Compostela. Informed consent was asked from patients receiving M-PC. M-PCs were treated with the Mirasol system according to the manufacturer's instructions. Thrombocytopenic patients were transfused according to the Spanish transfusion guidelines. Post-transfusion platelet counts were measured at 1 h and/or 24 h after transfusion. Post-transfusion surveillance of patients was maintained during the study. Results Data from 54 evaluable patients and 135 transfusions were analyzed. The mean age of patients was 58 years. The mean age of M-PC at transfusion was 3.6 days. The mean platelet dose was 3.7 × 1011. The transfusion responses measured as mean corrected count increment 1 h after transfusion (CCI1h) and CCI24h were 9,659 and 4,751, respectively. 65% of transfusions resulted in CCI1h values ≥ 7,500. 51% of transfusions resulted in CCI24h values ≥ 4,500. Conclusion The use of M-PC in the supportive treatment proved to be safe and effective for this cohort of thrombocytopenic patients.

  13. Bacterial contamination of platelet concentrates: results of a prospective multicenter study comparing pooled whole blood-derived platelets and apheresis platelets.

    PubMed

    Schrezenmeier, Hubert; Walther-Wenke, Gabriele; Müller, Thomas H; Weinauer, Franz; Younis, Adelheid; Holland-Letz, Tim; Geis, Gabriele; Asmus, Jens; Bauerfeind, Ursula; Burkhart, Jürgen; Deitenbeck, Robert; Förstemann, Elisabeth; Gebauer, Wolfgang; Höchsmann, Britta; Karakassopoulos, Apostolos; Liebscher, Ute-Maja; Sänger, Werner; Schmidt, Michael; Schunter, Friedrich; Sireis, Walid; Seifried, Erhard

    2007-04-01

    The GERMS Group initiated a prospective multicenter study to assess prevalence and nature of bacterial contamination of pooled buffy-coat platelet concentrates (PPCs) and apheresis platelet concentrates (APCs) by routine screening with a bacterial culture system. In nine centers overall, 52,243 platelet (PLT) concentrates (15,198 APCs, 37,045 PPCs) were analyzed by aerobic and anaerobic cultures (BacT/ALERT, bioMérieux). In 135 PLT concentrates (PCs; 0.26%), bacteria could be identified in the first culture (0.4% for APCs vs. 0.2% for PPCs; p < 0.001). In 37 (0.07%) of these PC units, the same bacteria strain could be identified in a second culture from the sample bag and/or the PC unit. The rate of confirmed-positive units did not differ significantly between APC (0.09%; 1/1169) and PPC units (0.06%; 1/1544). Bacteria from skin flora (Propionibacterium acnes, Staphylococcus epidermidis) were the most prevalent contaminants. Median times to first positive culture from start of incubation were 0.7 and 3.7 days in aerobic and anaerobic cultures for confirmed-positive units. With a "negative-to-date" issue strategy, most PC units (55%) had already been issued by time of the first positive culture. The rate of confirmed bacterial contamination of PC units was low. Nevertheless, clinicians must be aware of this risk. The risk of bacterial contamination does not warrant universal preference of APCs. It must be questioned whether routine bacterial screening by a culture method can sufficiently prevent contaminated products from being transfused due to the delay until a positive signal in the culture system and due to false-negative results.

  14. Freezing of Apheresis Platelet Concentrates in 6% Dimethyl Sulfoxide: The First Preliminary Study in Turkey

    PubMed Central

    Yılmaz, Soner; Çetinkaya, Rıza Aytaç; Eker, İbrahim; Ünlü, Aytekin; Uyanık, Metin; Tapan, Serkan; Pekoğlu, Ahmet; Pekel, Aysel; Erkmen, Birgül; Muşabak, Uğur; Yılmaz, Sebahattin; Avcı, İsmail Yaşar; Avcu, Ferit; Kürekçi, Emin; Eyigün, Can Polat

    2016-01-01

    Objective: Transfusion of platelet suspensions is an essential part of patient care for certain clinical indications. In this pioneering study in Turkey, we aimed to assess the in vitro hemostatic functions of platelets after cryopreservation. Materials and Methods: Seven units of platelet concentrates were obtained by apheresis. Each apheresis platelet concentrate (APC) was divided into 2 equal volumes and frozen with 6% dimethyl sulfoxide (DMSO). The 14 frozen units of APCs were kept at -80 °C for 1 day. APCs were thawed at 37 °C and diluted either with autologous plasma or 0.9% NaCl. The volume and residual numbers of leukocytes and platelets were tested in both before-freezing and post-thawing periods. Aggregation and thrombin generation tests were used to analyze the in vitro hemostatic functions of platelets. Flow-cytometric analysis was used to assess the presence of frozen treated platelets and their viability. Results: The residual number of leukocytes in both dilution groups was <1x106. The mean platelet recovery rate in the plasma-diluted group (88.1±9.5%) was higher than that in the 0.9% NaCl-diluted group (63±10%). These results were compatible with the European Directorate for the Quality of Medicines quality criteria. Expectedly, there was no aggregation response to platelet aggregation test. The mean thrombin generation potential of post-thaw APCs was higher in the plasma-diluted group (2411 nmol/L per minute) when compared to both the 0.9% NaCl-diluted group (1913 nmol/L per minute) and the before-freezing period (1681 nmol/L per minute). The flow-cytometric analysis results for the viability of APCs after cryopreservation were 94.9% and 96.6% in the plasma and 0.9% NaCl groups, respectively. Conclusion: Cryopreservation of platelets with 6% DMSO and storage at -80 °C increases their shelf life from 7 days to 2 years. Besides the increase in hemostatic functions of platelets, the cryopreservation process also does not affect their viability

  15. Freezing of Apheresis Platelet Concentrates in 6% Dimethyl Sulfoxide: The First Preliminary Study in Turkey.

    PubMed

    Yılmaz, Soner; Çetinkaya, Rıza Aytaç; Eker, İbrahim; Ünlü, Aytekin; Uyanık, Metin; Tapan, Serkan; Pekoğlu, Ahmet; Pekel, Aysel; Erkmen, Birgül; Muşabak, Uğur; Yılmaz, Sebahattin; Avcı, İsmail Yaşar; Avcu, Ferit; Kürekçi, Emin; Eyigün, Can Polat

    2016-03-05

    Transfusion of platelet suspensions is an essential part of patient care for certain clinical indications. In this pioneering study in Turkey, we aimed to assess the in vitro hemostatic functions of platelets after cryopreservation. Seven units of platelet concentrates were obtained by apheresis. Each apheresis platelet concentrate (APC) was divided into 2 equal volumes and frozen with 6% dimethyl sulfoxide (DMSO). The 14 frozen units of APCs were kept at -80 °C for 1 day. APCs were thawed at 37 °C and diluted either with autologous plasma or 0.9% NaCl. The volume and residual numbers of leukocytes and platelets were tested in both before-freezing and post-thawing periods. Aggregation and thrombin generation tests were used to analyze the in vitro hemostatic functions of platelets. Flow-cytometric analysis was used to assess the presence of frozen treated platelets and their viability. The residual number of leukocytes in both dilution groups was <1x106. The mean platelet recovery rate in the plasma-diluted group (88.1±9.5%) was higher than that in the 0.9% NaCl-diluted group (63±10%). These results were compatible with the European Directorate for the Quality of Medicines quality criteria. Expectedly, there was no aggregation response to platelet aggregation test. The mean thrombin generation potential of post-thaw APCs was higher in the plasma-diluted group (2411 nmol/L per minute) when compared to both the 0.9% NaCl-diluted group (1913 nmol/L per minute) and the before-freezing period (1681 nmol/L per minute). The flow-cytometric analysis results for the viability of APCs after cryopreservation were 94.9% and 96.6% in the plasma and 0.9% NaCl groups, respectively. Cryopreservation of platelets with 6% DMSO and storage at -80 °C increases their shelf life from 7 days to 2 years. Besides the increase in hemostatic functions of platelets, the cryopreservation process also does not affect their viability rates.

  16. Pathogen inactivation treatment of plasma and platelet concentrates and their predicted functionality in massive transfusion protocols.

    PubMed

    Arbaeen, Ahmad F; Schubert, Peter; Serrano, Katherine; Carter, Cedric J; Culibrk, Brankica; Devine, Dana V

    2017-05-01

    Trauma transfusion packages for hemorrhage control consist of red blood cells, plasma, and platelets at a set ratio. Although pathogen reduction improves the transfusion safety of platelet and plasma units, there is an associated reduction in quality. This study aimed to investigate the impact of riboflavin/ultraviolet light-treated plasma or platelets in transfusion trauma packages composed of red blood cell, plasma, and platelet units in a ratio of 1:1:1 in vitro by modeling transfusion scenarios for trauma patients and assessing function by rotational thromboelastometry. Pathogen-reduced or untreated plasma and buffy coat platelet concentrate units produced in plasma were used in different combinations with red blood cells in trauma transfusion packages. After reconstitution of these packages with hemodiluted blood, the hemostatic functionality was analyzed by rotational thromboelastometry. Hemostatic profiles of pathogen-inactivated buffy coat platelet concentrate and plasma indicated decreased activity compared with their respective controls. Reconstitution of hemodiluted blood (hematocrit = 20%) with packages that contained treated or nontreated components resulted in increased alpha and maximum clot firmness and enhanced clot-formation time. Simulating transfusion scenarios based on 30% blood replacement with a transfusion trauma package resulted in a nonsignificant difference in rotational thromboelastometry parameters between packages containing treated and nontreated blood components (p ≥ 0.05). Effects of pathogen inactivation treatment were evident when the trauma package percentage was 50% or greater and contained both pathogen inactivation-treated plasma and buffy coat platelet concentrate. Rotational thromboelastometry investigations suggest that there is relatively little impact of pathogen inactivation treatment on whole blood clot formation unless large amounts of treated components are used. © 2017 AABB.

  17. Preparation of small volume, leuko and erythrocyte very poor platelet concentrates.

    PubMed

    Valbonesi, M; Angelini, G; Malfanti, L; Lercari, G; Fella, M; Calderisi, S; Anselmo, A; Balistreri, M

    1986-05-01

    Recently developed automated discontinuous flow centrifuge (DFC) separators can produce leuko- and erythrocyte-poor platelet concentrates (PC). According to general experience with these machines it is difficult to obtain more than 4 X 10(11) platelets, though a second program set up by Coffe et al. appears to produce PC containing approximately 5 X 10(11) platelets suspended in a plasma volume of 390 ml. At our center we employed a new Dideco cell separator equipped with the surge pump and a technique developed for the production of small volume, RBC and WBC-very poor PC. In 60 routine procedures we obtained the following results: mean processing time 87 +/- 11 minutes; final volume of PC 136 +/- 19 ml, with a mean platelet yield of 5.21 X 10(11) platelets. WBC contamination was 1.8 X 10(8) (93% lymphocytes) and RBC were 3.1 X 10(8). Plasma volume as well as WBC and RBC contamination were reduced by recirculating PC after the 6th pass. The demand for single donor platelet concentrates (PC) is increasing progressively. Recently developed automated cell separators can produce leukocyte (WBC) and erythrocyte (RBC) poor PC. With these machines it may be difficult to obtain PC containing at least 4 X 10(11) platelets and less than 1 X 10(9) leukocytes (1, 2, 3) since donor variables such as hematocrit, precounts, buffy coat formation and initial plasma light transmission are of paramount importance for the efficiency of the program. At our center a prototype discontinuous flow centrifuge (DFC) cell separator equipped with the surge pump was studied.(ABSTRACT TRUNCATED AT 250 WORDS)

  18. Evolution, current status and advances in application of platelet concentrate in periodontics and implantology.

    PubMed

    Agrawal, Amit Arvind

    2017-05-16

    Platelet concentrates (PC) [platelet-rich plasma (PRP) and platelet-rich fibrin (PRF)] are frequently used for surgical procedures in medical and dental fields, particularly in oral and maxillofacial surgery, plastic surgery and sports medicine. The objective of all these technologies is to extract all the elements from a blood sample that could be used to improve healing and promote tissue regeneration. Although leukocyte rich and leukocyte poor PRP's have their own place in literature, the importance of non-platelet components in a platelet concentrate remains a mystery. PC have come a long way since its first appearance in 1954 to the T-PRF, A-PRF and i-PRF introduced recently. These PC find varied applications successfully in periodontics and implant dentistry as well. However, the technique of preparation, standing time, transfer process, temperature of centrifuge, vibration, etc., are the various factors for the mixed results reported in the literature. Until the introduction of a proper classification of terminologies, the PC were known by different names in different countries and by different commercial companies which also created a lot of confusion. This review intends to clarify all these confusion by briefing the exact evolution of PC, their preparation techniques, recent advances and their various clinical and technical aspects and applications.

  19. Evolution, current status and advances in application of platelet concentrate in periodontics and implantology

    PubMed Central

    Agrawal, Amit Arvind

    2017-01-01

    Platelet concentrates (PC) [platelet-rich plasma (PRP) and platelet-rich fibrin (PRF)] are frequently used for surgical procedures in medical and dental fields, particularly in oral and maxillofacial surgery, plastic surgery and sports medicine. The objective of all these technologies is to extract all the elements from a blood sample that could be used to improve healing and promote tissue regeneration. Although leukocyte rich and leukocyte poor PRP’s have their own place in literature, the importance of non-platelet components in a platelet concentrate remains a mystery. PC have come a long way since its first appearance in 1954 to the T-PRF, A-PRF and i-PRF introduced recently. These PC find varied applications successfully in periodontics and implant dentistry as well. However, the technique of preparation, standing time, transfer process, temperature of centrifuge, vibration, etc., are the various factors for the mixed results reported in the literature. Until the introduction of a proper classification of terminologies, the PC were known by different names in different countries and by different commercial companies which also created a lot of confusion. This review intends to clarify all these confusion by briefing the exact evolution of PC, their preparation techniques, recent advances and their various clinical and technical aspects and applications. PMID:28560233

  20. Multicentre standardisation of a clinical grade procedure for the preparation of allogeneic platelet concentrates from umbilical cord blood

    PubMed Central

    Rebulla, Paolo; Pupella, Simonetta; Santodirocco, Michele; Greppi, Noemi; Villanova, Ida; Buzzi, Marina; De Fazio, Nicola; Grazzini, Giuliano

    2016-01-01

    Background In addition to a largely prevalent use for bleeding prophylaxis, platelet concentrates from adult blood have also been used for many years to prepare platelet gels for the repair of topical skin ulcers. Platelet gel can be obtained by activation of fresh, cryopreserved, autologous or allogeneic platelet concentrates with calcium gluconate, thrombin and/or batroxobin. The high content of tissue regenerative factors in cord blood platelets and the widespread availability of allogeneic cord blood units generously donated for haematopoietic transplant but unsuitable for this use solely because of low haematopoietic stem cell content prompted us to develop a national programme to standardise the production of allogeneic cryopreserved cord blood platelet concentrates (CBPC) suitable for later preparation of clinical-grade cord blood platelet gel. Materials and methods Cord blood units collected at public banks with total nucleated cell counts <1.5×109, platelet count >150×109/L and volume >50 mL, underwent soft centrifugation within 48 hours of collection. Platelet-rich plasma was centrifuged at high speed to obtain a CBPC with target platelet concentration of 800–1,200×109/L, which was cryopreserved, without cryoprotectant, below −40 °C. Results During 14 months, 13 banks produced 1,080 CBPC with mean (± standard deviation) volume of 11.4±4.4 mL and platelet concentration of 1,003±229×109/L. Total platelet count per CBPC was 11.3±4.9×109. Platelet recovery from cord blood was 47.7±17.8%. About one-third of cord blood units donated for haematopoietic transplant could meet the requirements for preparation of CBPC. The cost of preparation was € 160.92/CBPC. About 2 hours were needed for one technician to prepare four CBPC. Discussion This study yielded valuable scientific and operational information regarding the development of clinical trials using allogeneic CBPC. PMID:26509822

  1. [Transfusions of rhesus-incompatible platelet concentrates in Rouen University Hospital: procedures and consequences].

    PubMed

    Chamouni, P; Josset, V; Bastit, D; Tavolacci, M P; Lenain, P; Varin, R; Czernichow, P

    2005-10-01

    Guidelines for distribution and use of blood products have been established for both blood transfusion institution and hospitals, in particular for the use of Rh (D)-incompatible platelet concentrates. The aim of this study was to evaluate: 1) the rate of attribution for the Rh (D)-incompatible platelets concentrates, 2) the immunisation prophylaxis practices, 3) the immunological consequences using short and medium term follow-up of transfused patients. Patients with Rh (D)-incompatible platelets concentrate administered during the year 2003 at Rouen University Hospital were retrospectively selected. Patients on transfusion were described. The relationship of various factors with the injection as well as the appearance of allo-immunization was statistically tested. During a year, 280 Rh (D)-incompatible platelets concentrates were administered to 67 patients. Immunisation prophylaxis by injection of Ig anti-D was not systematically performed. Four immunizations in the Rhesus group system were identified: 2 against D antigen (Ag), 1 against E Ag and 1 against C Ag. Immunisations against D Ag occurred for two younger women considered as immunodeficient. Immunization prophylaxis was more frequent in poly-transfused patients. However no difference was observed for the other factors. Compatibility concerning Rhesus (D) is not always possible. The immunization against red cells persists, in particular against the antigens of the Rhesus group system and moreover for the immunodeficient patients. Recommendations for immunization prophylaxis by injection of specific anti-D immune-globulin (Ig) could be reconsidered.

  2. Coagulation factors bound to procoagulant platelets concentrate in cap structures to promote clotting.

    PubMed

    Podoplelova, Nadezhda A; Sveshnikova, Anastasia N; Kotova, Yana N; Eckly, Anita; Receveur, Nicolas; Nechipurenko, Dmitry Yu; Obydennyi, Sergey I; Kireev, Igor I; Gachet, Christian; Ataullakhanov, Fazly I; Mangin, Pierre H; Panteleev, Mikhail A

    2016-09-29

    Binding of coagulation factors to phosphatidylserine (PS)-exposing procoagulant-activated platelets followed by formation of the membrane-dependent enzyme complexes is critical for blood coagulation. Procoagulant platelets formed upon strong platelet stimulation, usually with thrombin plus collagen, are large "balloons" with a small (∼1 μm radius) "cap"-like convex region that is enriched with adhesive proteins. Spatial distribution of blood coagulation factors on the surface of procoagulant platelets was investigated using confocal microscopy. All of them, including factors IXa (FIXa), FXa/FX, FVa, FVIII, prothrombin, and PS-sensitive marker Annexin V were distributed nonhomogeneously: they were primarily localized in the "cap," where their mean concentration was by at least an order of magnitude, higher than on the "balloon." Assembly of intrinsic tenase on liposomes with various PS densities while keeping the PS content constant demonstrated that such enrichment can accelerate this reaction by 2 orders of magnitude. The mechanisms of such acceleration were investigated using a 3-dimensional computer simulation model of intrinsic tenase based on these data. Transmission electron microscopy and focal ion beam-scanning electron microscopy with Annexin V immunogold-labeling revealed a complex organization of the "caps." In platelet thrombi formed in whole blood on collagen under arterial shear conditions, ubiquitous "caps" with increased Annexin V, FX, and FXa binding were observed, indicating relevance of this mechanism for surface-attached platelets under physiological flow. These results reveal an essential heterogeneity in the surface distribution of major coagulation factors on the surface of procoagulant platelets and suggest its importance in promoting membrane-dependent coagulation reactions.

  3. Second-generation Platelet Concentrate (Platelet-rich Fibrin) as a Scaffold in Regenerative Endodontics: A Case Series.

    PubMed

    Bakhtiar, Hengameh; Esmaeili, Shahram; Fakhr Tabatabayi, Setareh; Ellini, Mohammad Reza; Nekoofar, Mohammad Hossein; Dummer, Paul M H

    2017-03-01

    The purpose of this case series was to report the clinical and radiographic results of a pulp regenerative procedure using platelet-rich fibrin (PRF), a second-generation platelet concentrate, in immature teeth with necrotic pulps. Root canal revascularization using PRF was performed on 4 immature teeth with necrotic pulps. After access cavity preparation, the root canals were irrigated with low concentration sodium hypochlorite solution (1.5% sodium hypochlorite [20 mL/canal, 5 minutes]) and then irrigated with saline (20 mL/canal, 5 minutes). Equal proportions (167 mg) of ciprofloxacin, metronidazole, and cefaclor were mixed and diluted to a final concentration of 1 g/mL. Finally, the canal was sealed with 3-4 mm of a temporary restorative material, and patients were dismissed for 2 to 3 weeks. At the second appointment, 9 mL of the patient's whole blood was obtained and centrifuged to prepare a PRF clot. Canals were irrigated with 17% EDTA, and a sharp spreader was inserted beyond the apex. Then, the PRF clot was placed inside the root canals, and Biodentine (Septodont, Saint-Maur, France) was placed directly over the PRF. The teeth were restored permanently with glass ionomer cement and composite resin. Clinical examinations revealed that all cases were asymptomatic at the recall appointments at 1, 3, 6, 12, and 18 months. Radiographs revealed resolution of the periapical lesions, further root development, and apical closure in all cases. On the basis of the short-term results up to 12 months, PRF clots acted as successful scaffolds for the regeneration of pulpal contents in immature teeth with necrotic pulps. Copyright © 2016 American Association of Endodontists. All rights reserved.

  4. Von Willebrand factor availability in platelet concentrates stored for 5 days.

    PubMed

    Cesar, J M; García-Avello, A; Monteagudo, J; Espinosa, J I; Lodos, J C; Castillo, R; Navarro, J L

    1994-02-01

    Von Willebrand factor (vWF) availability was assessed in platelet concentrates (PCs). After 5 days of storage, 82 +/- 9% of basal levels of ristocetin cofactor activity (vWF:RCo) remained in PCs. vWF antigen (vWF:Ag) increased up to 166 +/- 38% (P < 0.05) in the same period. Autoradiograph pattern of vW:Ag showed an increase in low molecular weight multimers, and fast migrating multimeric forms were visualized by crossed immunoelectrophoresis on day 5. Studies carried out in platelet free plasma stored as PCs showed similar changes in vWF:RCo but increments in vWF:Ag were not detected. These data indicate that PCs maintain vWF:RCo levels of clinical value even after 5 days of storage and suggest that vWF comes out from platelets to plasma during storage.

  5. Platelet concentrates for topical use: bedside device and blood transfusion technology. Quality and versatility.

    PubMed

    Borzini, Piero; Balbo, Valeria; Mazzucco, Laura

    2012-06-01

    More or less after a decade of experimental and pioneering manual procedures to prepare platelet-rich plasma (PRP) for topical use, several portable and bedside devices were made available to prepare the PRP at the point-of-care. This technical opportunity increased the number of patients who got access to the treatment with autologous PRP and PRP-gel. Since topical treatment of tissue with PRP and PRP-gel was restricted to autologous preparation, blood transfusion centers that professionally prepare donor-derived platelet concentrates were not able to cover the overwhelming request for autologous PRP supply. Principally for logistic and organization reasons blood transfusion centers usually fail the challenge of prompt delivery of PRP to the physician over large territory. Nevertheless the blood bank production of platelet concentrates is associated with high standardization and quality controls not achievable from bedside and portable devices. Furthermore it easy to demonstrate that high-volume blood bank-produced platelet concentrates are less expensive than low-volume PRP produced by portable and bedside devices. Taking also in consideration the ever-increasing safety of the blood components, the relationship between bedside device-produced and blood-bank-produced PRP might be reconsidered. Here we discuss this topic concluding that the variety of sources of PRP production is an opportunity for versatility and that, ultimately, versatility is an opportunity for the patient's care.

  6. Variance of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) concentrations in activated, concentrated platelets from healthy male donors

    PubMed Central

    2014-01-01

    Background The use of autologous blood concentrates, such as activated, concentrated platelets, in orthopaedic clinical applications has had mixed results. Research on this topic has focused on growth factors and cytokines, with little directed towards matrix metalloproteinases (MMPs) which are involved in post-wound tissue remodeling. Methods In this study, the authors measured the levels of MMP-2, MMP-9 and a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13), in activated platelets derived from blood of healthy, male volunteers (n = 92), 19 to 60 years old. The levels of the natural inhibitors of these proteases, tissue inhibitor of metalloproteinase 1 (TIMP-1), TIMP-2 and TIMP-4 were also assessed. Results Notably, there was no significant change in concentration with age in four of six targets tested. However, TIMP-2 and TIMP-4 demonstrated a statistically significant increase in concentration for subjects older than 30 years of age compared to those 30 years and younger (P = 0.04 and P = 0.04, respectively). Conclusion TIMP-2 and TIMP-4 are global inhibitors of MMPs, including MMP-2 (Gelatinase A). MMP-2 targets native collagens, gelatin and elastin to remodel the extracellular matrix during wound healing. A decreased availability of pharmacologically active MMP-2 may diminish the effectiveness of the use of activated, concentrated platelets from older patients, and may also contribute to longer healing times in this population. PMID:24766991

  7. Effect of membrane protein concentration on binding of /sup 3/H-imipramine in human platelets

    SciTech Connect

    Barkai, A.I.; Kowalik, S.; Baron, M.

    1985-02-01

    Binding of /sup 3/H-imipramine to platelet membranes has been implicated as a marker for depression. Comparing /sup 3/H-IMI binding between depressed patients and normal subjects we observed an increase in the dissociation constant Kd with increasing membrane protein. This phenomenon was studied more rigorously in five normal subjects. Platelet membranes were prepared and adjusted to four concentrations of protein ranging from 100 to 800 micrograms/ml. The /sup 3/H-IMI binding parameters of maximum binding sites number (Bmax) and Kd were obtained by Scatchard analysis at each membrane concentration. A positive linear relationship was found between K/sub d/ values and the concentration of membrane protein in the assay, but no change was observed in Bmax. The variability in Kd values reported in the literature may be accounted for in part by the different concentrations of membrane protein used in various studies.

  8. [Effects of use of riboflavin and ultraviolet light for pathogen inactivation on quality of platelet concentrates].

    PubMed

    Stanojković, Zoran; Antić, Ana; Stojanović, Miodrag

    2011-06-01

    Pathogen inactivation in blood and blood products is one of the major means to achieve a zero risk blood supply and improve transfusion safety. Riboflavin (vitamin B2) activated by ultraviolet (UV) light, produces active oxygen which damages cell membrane and prevents replication of the carrier of diseases (viruses, bacteria, protozoa) in all blood products. The aim of this study was to establish the influence of the process of pathogens photoinactivation using riboflavin and UV rays on the biochemical and functional characteristics of platelet concentrates prepared from "buffy coat". The examination included 80 platelet concentrates prepared from "buffy coat", which was separated from whole blood donated by voluntary blood donors around 6 hours from the moment of collection. Concentrates were pooled, filtered and separated unton two groups: one consisted of 10 control units and the other of 10 examined units (pooled platelet concentrates). Examined units of the platelets were treated by riboflavin (35 mL) and UV rays (6.24 J/mL, 265-370 nm) on Mirasol aparature (Caridian BCT Biotechnologies, USA) in approximate duration of 6 min. A total of 35 mL of saline solution was added to the control units. The samples for examining were taken from the control and examined units initially (K0, I0), after the addition of saline (K1) and riboflavin (I1), after illumination (I2), first day of storage (K3, I3) and the fifth day of storage (K4, I4). The following parameters were measured: platelet count and platelet yield, residual erythrocyte and leukocyte count, pH, pO2, pCO2 and bacterial contamination. All the measured parameters showed a statistically significant decrease comparing to K0 and I0; all the results of the first day of platelet storage showed statistically significant decrease comparing to K1 and I1, and all the results of the fifth day of platelet storage (K4, I4) showed a statistically significant decrease comparing to K1 and K3 and to I1 and I3. There was no

  9. Extended Storage of Pathogen-Reduced Platelet Concentrates (PRECON)

    DTIC Science & Technology

    2014-10-01

    standard Terumo plt storage bag (PVC plastic ). However, Haemonetics apheresis plts can be stored for 13 days in Haemonetics bags (CLX plastic ). Plt...concentrates will first be stored for 6 days in 65% PAS/35% plasma using our standard Terumo plt storage bag (PVC plastic ) If FDA criteria are met...Preliminary steps to this ultimate goal are to determine the best storage bag and the optimum PAS to plasma ratio to utilize in our Mirasol experiments. Once

  10. [Platelet concentrates from whole-blood donations (buffy-coat) or apheresis: which one to use?].

    PubMed

    Lozano, María Luisa; Rivera, José; Vicente, Vicente

    2012-05-05

    Platelet concentrates (PCs) prepared either from whole-blood donations by the buffy-coat method (BC), or by plateletpheresis are indicated to prevent or treat acute hemorrhage secondary to thrombocytopenia, and there is an ongoing debate about which platelet product should be used. Usage of each of these two products is highly heterogeneous among countries and individual institutions, ranging from 10 to 90%, with a 50:50 ratio in Europe. In comparison of pooled platelets prepared by the BC method and apheresis PCs, data suggest similar efficacy of the products. Regarding recipients' adverse reactions, there is no advantage for apheresis concentrates. From the donor's point of view, evidence favours using the abundance of platelets available from whole-blood donation. As residual viral transmission risk continues to fall, the advantage of apheresis products related to the decrease to donor exposure lessens. While the cost-effectiveness of apheresis products is comparable to that of other accepted blood safety interventions, in case of emerging pathogens, probably pathogen inactivation of pooled BC PCs would be a more desirable strategy.

  11. The inability of tegaserod to affect platelet aggregation and coronary artery tone at supratherapeutic concentrations.

    PubMed

    Higgins, Deborah L; Ero, Mike P; Loeb, Michelle; Kersey, Kathryn; Hopkins, Alan; Beattie, David T

    2012-01-01

    In 2007, the results from a meta analysis of 29 clinical studies indicated that tegaserod (Zelnorm®), a 5-hydroxytryptamine(4) (5-HT(4)) receptor agonist with gastrointestinal prokinetic activity, was associated with an increased incidence of cardiovascular ischemic events, resulting in its withdrawal from many markets around the world. Stimulation of platelet aggregation has been proposed to explain the phenomenon. However, data from recent epidemiological studies have suggested that there is no correlation between tegaserod use and the incidence of cardiovascular ischemia. In this study, the influence of tegaserod, at concentrations up to tenfold higher than the total plasma C (max) for the 6 mg clinical dose, has been investigated on platelet aggregation under standard conditions with platelet-rich plasma (PRP) obtained from healthy human subjects. Additionally, the influence of tegaserod on coronary artery tone was evaluated as an alternative pro-ischemic mechanism. The positive control, thrombopoietin, but not tegaserod, demonstrated a statistically significant increase in platelet aggregation using the same PRP samples with either adenosine diphosphate (ADP) or ADP plus 5-HT as an aggregation agonist. Tegaserod had no contractile activity in either porcine or human isolated coronary artery preparations, and only a small and variable response in canine coronary arteries at concentrations higher than those achieved clinically. Taken together, these studies do not identify a mechanism for the ischemic events that have been attributed to tegaserod in humans.

  12. Increasing susceptibility of nitric oxide-mediated inhibitory platelet signaling during storage of apheresis-derived platelet concentrates.

    PubMed

    Kobsar, Anna; Klinker, Erdwine; Kuhn, Sabine; Koessler, Angela; Yilmaz, Pinar; Boeck, Markus; Koessler, Juergen

    2014-07-01

    Storage of platelets (PLTs) affects PLT integrity and functionality, a process named the PLT storage lesion. Normal PLT function essentially depends on the balanced interaction of activating and inhibitory signaling pathways. As there are poor data on the alterations of inhibitory signaling during storage of PLT concentrates, this study investigates the modulation capability of the cyclic nucleotide-mediated inhibitory pathways by use of the nitric oxide donor diethylamine diazenium diolate (DEA/NO). PLTs were obtained from whole blood (WB) and from apheresis-derived PLT concentrates (APCs) stored for 0, 2, and 5 days. Vasodilator-stimulated phosphoprotein (VASP) phosphorylation, cyclic nucleotide concentrations, fibrinogen binding, and agonist-induced aggregation were measured without or after stimulation with DEA/NO. DEA/NO-induced VASP phosphorylation was significantly higher in PLTs from APCs on Days 2 and 5 compared to WB, conditioned by a stronger increase of cyclic guanosine monophosphate (cGMP), but not cyclic adenosine monophosphate (cAMP), in stored PLTs. A quantity of 5 nmol/L DEA/NO neither influenced thrombin receptor activator peptide 6 and collagen-induced aggregation nor fibrinogen binding in freshly collected PLTs, whereas it significantly inhibited both in stored PLTs. Stored PLTs showed an impairment of intracellular cGMP regulation, resulting in exceeding inhibition of agonist-induced aggregation and fibrinogen binding in the course of storage. The observed effects could be an important mechanism contributing to the storage lesion with reduced activating potential of PLTs. © 2014 AABB.

  13. Evaluation of platelet function using the in vitro bleeding time and corrected count increment of transfused platelets. Comparison between platelet concentrates derived from pooled buffy coates and apheresis.

    PubMed

    Eriksson, L; Kristensen, J; Olsson, K; Bring, J; Högman, C F

    1996-01-01

    The functional capacity of transfused platelets was evaluated with in vitro bleeding time (IVBT) and corrected count increment (CCI) in order to compare platelet concentrates (PCs) derived from pooled buffy coats (BC-PCs) with PCs collected by apheresis (A-PCs). The suspension medium in the BC-PCs was 30% CPD plasma and 70% of an additive solution (containing sodium and potassium chloride, sodium citrate and phosphate, mannitol), and in the A-PCs the medium was 100% CPD plasma. IVBT was evaluated using a Thrombostat 4000/2. BC-PC and A-PC were transfused 57 and 41 times, respectively to 36 patients with chemotherapy-induced thrombocytopenia. PCs transfused within 2 days of donation were considered fresh, and those transfused within 3-5 days were considered stored. IVBT was determined before, as well as 10-30 min and 24 h after transfusion; CCI was determined 10-30 min and 24 h after transfusion. The median pretransfusion IVBT value was 486 s. It was measurable in 21 of 98 (21%) of the transfusions, i.e. below the cutoff limit of 486 s. Ten to 30 min after transfusion, the IVBT showed a measurable reduction in 90% of the transfusions with fresh BC-PCs, 92% of those with fresh a-PCs, 63% of those with stored BC-PCs and 79% of those with stored A-PCs. After 24 h, the corresponding values were 63% for fresh BC-PCs, 50% for fresh A-PCs, 26% for stored BC-PCs and 38% for stored A-PCs. The median value of CCI 10-30 min after transfusion was 20 for fresh BC-PCs, 17 for fresh A-PCs, 16 for stored BC-PCs and 14 for stored A-PCs. The difference in IVBT between fresh and stored BC-PCs was significant (p = 0.032), unlike that between fresh and stored A-PC. After 24 h the corresponding values were 7 for fresh BC-PCs, 4 for fresh A-PCs, 4 for stored BC-PCs and 3 for stored A-PCs. When all transfusions with fresh PCs (BC-PCs + A-PCs) were compared with all transfusions with stored PCs, a statistical difference was demonstrated in both CCI (p = 0.027) and IVBT (p = 0.043). Spearman

  14. The role of bicarbonate in platelet additive solution for apheresis platelet concentrates stored with low residual plasma.

    PubMed

    Radwanski, Katherine; Min, Kyungyoon

    2013-03-01

    Complex platelet additive solutions (PASs) are required to store platelet (PLT) concentrates with plasma levels below 30%. Previously, apheresis PLTs stored with 5% plasma in acetate- and bicarbonate-containing PAS maintained stable pH and bicarbonate levels during 7-day storage. Due to this observation, the necessity of added bicarbonate in PAS was investigated and whether the concurrent increase in PAS pH after bicarbonate addition had any effect on PLT storage. Apheresis PLTs were stored in 5% plasma-95% high- or low-pH PAS, with or without bicarbonate (n=10 per arm). Bicarbonate PAS PLTs were paired and nonbicarbonate PAS PLTs were paired (split from same double-dose collection). PLTs were evaluated for in vitro variables on Days 1 and 7 and up to Day 14 if the Day 7 pH was higher than 6.2. PLT pH was maintained above 7.3 to Day 14 in bicarbonate PAS PLTs while pH failures below 6.2 were observed in 4 of 10 and 2 of 10 units on Day 7 in low- and high-pH nonbicarbonate PAS arms, respectively. Day 7 in vitro variables in nonbicarbonate PAS PLTs with pH values of higher than 6.2 were comparable to Day 7 variables in bicarbonate PAS PLTs. The pH of bicarbonate PAS did have a small effect on pH and bicarbonate levels in PLT units, but did not have an effect on functional variables and metabolism. Bicarbonate was not required to maintain in vitro PLT function in 5% plasma-95% PAS, but was required as a pH buffer and increased PAS pH did not significantly contribute to this effect. © 2012 American Association of Blood Banks.

  15. Effect of platelet rich plasma gel in a physiologically relevant platelet concentration on wounds in persons with spinal cord injury.

    PubMed

    Rappl, Laurie M

    2011-04-01

    The objective of the study was to investigate the use of a 1·3 times normal platelet concentration platelet-rich plasma (PRP) gel to move chronic wounds towards healing in persons with spinal cord injury (SCI). The study design was a case series of 20 persons with SCI with non healing wounds. The outcome measures were, in wound area, volume, undermining and sinus tracts/tunnels (ST/Ts), calculated average, (i) percent of change from baseline, (ii) change per day from baseline, (iii) number of treatments and (iv) number of weeks. In a mean of 4·0, after treatments over 3·4 weeks, the wounds closed on an average of 47·9% in area and 56·0% in volume. Undermining closed on an average of 31·4% using 3·5 treatments over 2·6 weeks. ST/Ts closed on an average of 26·1% after 2·3 treatments over 1·5 weeks. Clinical relevance by percent of positive responders and their response: in area, 90·0% of the subjects responded positively, the average reduction was 53·8%. In volume, 90·0% responded, with an average reduction of 67·3%. Of four subjects with undermining, 75% closed 47·0% on average. Of the three with ST/Ts, 100% closed 26·1% on average. Average haemoglobin and haematocrit levels were below normal. To conclude, 1·3× PRP gel appears to progress chronic, non healing wounds in SCI patients into the granulation phase of healing quickly. Review of the literature shows these results may not be applied to all PRP preparations. © 2011 The Author. © 2011 Blackwell Publishing Ltd and Medicalhelplines.com Inc.

  16. Bacterial screening of platelet concentrates on day 2 and 3 with flow cytometry: the optimal sampling time point?

    PubMed Central

    Vollmer, Tanja; Schottstedt, Volkmar; Bux, Juergen; Walther-Wenke, Gabriele; Knabbe, Cornelius; Dreier, Jens

    2014-01-01

    Background There is growing concern on the residual risk of bacterial contamination of platelet concentrates in Germany, despite the reduction of the shelf-life of these concentrates and the introduction of bacterial screening. In this study, the applicability of the BactiFlow flow cytometric assay for bacterial screening of platelet concentrates on day 2 or 3 of their shelf-life was assessed in two German blood services. The results were used to evaluate currently implemented or newly discussed screening strategies. Materials and methods Two thousand and ten apheresis platelet concentrates were tested on day 2 or day 3 after donation using BactiFlow flow cytometry. Reactive samples were confirmed by the BacT/Alert culture system. Results Twenty-four of the 2,100 platelet concentrates tested were reactive in the first test by BactiFlow. Of these 24 platelet concentrates, 12 were false-positive and the other 12 were initially reactive. None of the microbiological cultures of the initially reactive samples was positive. Parallel examination of 1,026 platelet concentrates by culture revealed three positive platelet concentrates with bacteria detected only in the anaerobic culture bottle and identified as Staphylococcus species. Two platelet concentrates were confirmed positive for Staphylcoccus epidermidis by culture. Retrospective analysis of the growth kinetics of the bacteria indicated that the bacterial titres were most likely below the diagnostic sensitivity of the BactiFlow assay (<300 CFU/mL) and probably had no transfusion relevance. Conclusions The BactiFlow assay is very convenient for bacterial screening of platelet concentrates independently of the testing day and the screening strategy. Although the optimal screening strategy could not be defined, this study provides further data to help achieve this goal. PMID:24887230

  17. Bacterial screening of platelet concentrates on day 2 and 3 with flow cytometry: the optimal sampling time point?

    PubMed

    Vollmer, Tanja; Schottstedt, Volkmar; Bux, Juergen; Walther-Wenke, Gabriele; Knabbe, Cornelius; Dreier, Jens

    2014-07-01

    There is growing concern on the residual risk of bacterial contamination of platelet concentrates in Germany, despite the reduction of the shelf-life of these concentrates and the introduction of bacterial screening. In this study, the applicability of the BactiFlow flow cytometric assay for bacterial screening of platelet concentrates on day 2 or 3 of their shelf-life was assessed in two German blood services. The results were used to evaluate currently implemented or newly discussed screening strategies. Two thousand and ten apheresis platelet concentrates were tested on day 2 or day 3 after donation using BactiFlow flow cytometry. Reactive samples were confirmed by the BacT/Alert culture system. Twenty-four of the 2,100 platelet concentrates tested were reactive in the first test by BactiFlow. Of these 24 platelet concentrates, 12 were false-positive and the other 12 were initially reactive. None of the microbiological cultures of the initially reactive samples was positive. Parallel examination of 1,026 platelet concentrates by culture revealed three positive platelet concentrates with bacteria detected only in the anaerobic culture bottle and identified as Staphylococcus species. Two platelet concentrates were confirmed positive for Staphylcoccus epidermidis by culture. Retrospective analysis of the growth kinetics of the bacteria indicated that the bacterial titres were most likely below the diagnostic sensitivity of the BactiFlow assay (<300 CFU/mL) and probably had no transfusion relevance. The BactiFlow assay is very convenient for bacterial screening of platelet concentrates independently of the testing day and the screening strategy. Although the optimal screening strategy could not be defined, this study provides further data to help achieve this goal.

  18. Effect of swirling flow on platelet concentration distribution in small-caliber artificial grafts and end-to-end anastomoses

    NASA Astrophysics Data System (ADS)

    Zhan, Fan; Fan, Yu-Bo; Deng, Xiao-Yan

    2011-10-01

    Platelet concentration near the blood vessel wall is one of the major factors in the adhesion of platelets to the wall. In our previous studies, it was found that swirling flows could suppress platelet adhesion in small-caliber artificial grafts and end-to-end anastomoses. In order to better understand the beneficial effect of the swirling flow, we numerically analyzed the near-wall concentration distribution of platelets in a straight tube and a sudden tubular expansion tube under both swirling flow and normal flow conditions. The numerical models were created based on our previous experimental studies. The simulation results revealed that when compared with the normal flow, the swirling flow could significantly reduce the near-wall concentration of platelets in both the straight tube and the expansion tube. The present numerical study therefore indicates that the reduction in platelet adhesion under swirling flow conditions in small-caliber arterial grafts, or in end-to-end anastomoses as observed in our previous experimental study, was possibly through a mechanism of platelet transport, in which the swirling flow reduced the near-wall concentration of platelets.

  19. PLATELET CONCENTRATES PREPARED AFTER A 20 TO 24 HOUR HOLD OF THE WHOLE BLOOD AT 22°C

    PubMed Central

    Slichter, Sherrill J.; Corson, Jill; Jones, Mary Kay; Christoffel, Todd; Pellham, Esther; Bolgiano, Doug

    2012-01-01

    Background The FDA requires that red cells must be refrigerated within 8 hours of whole blood collection. Longer storage of whole blood at 22°C before component preparation would have many advantages. Study Design And Methods Two methods of holding whole blood for 20 to 24 hours at room temperature were evaluated; i.e., refrigerated plates or a 23°C incubator. After extended whole blood storage, platelet concentrates were prepared from platelet-rich-plasma on day 1 post-donation, and the platelets were stored for 6 more days. On day 7 of platelet storage, blood was drawn from each subject to prepare fresh platelets. The stored and fresh platelets were radiolabeled and transfused into their donor. Results Eleven subjects’ whole blood was stored using refrigerated Compocool plates and 10 using an incubator. Post-storage platelet recoveries averaged 47 ± 13% versus 53 ± 11% and survivals averaged 4.6 ± 1.7 days versus 4.7 ± 0.9 days for Compocool versus incubator storage, respectively (p=N.S.). Using all results, post-storage platelet recoveries averaged 75 ± 10% of fresh and survivals 57 ± 13% of fresh; platelet recoveries met FDA guidelines for post-storage platelet viability but not survivals. Conclusion Seven-day post-storage platelet viability is comparable when whole blood is stored for 22 ± 2 hour at 22°C using either refrigerated plates or an incubator to maintain temperature prior to preparing platelet concentrates. PMID:22320682

  20. Pretreatment platelet 5-HT concentration predicts the short-term response to paroxetine in major depression. Grupo de Trastornos Afectivos.

    PubMed

    Figueras, G; Pérez, V; San Martino, O; Alvarez, E; Artigas, F

    1999-08-15

    A previous retrospective study revealed that a high pretreatment platelet serotonin (5-HT) concentration was associated with a low response to serotonergic antidepressants in drug-free major depressives. We have examined such a relationship in depressive patients treated with paroxetine. Seventy-four drug-free major depressives (DSM-IV) were admitted to the study. Clinical ratings were performed and blood was drawn prior to the initiation of treatment and after 4 weeks of paroxetine (20 mg/day). The concentrations of 5-HT, 5-hydroxyindoleacetic acid, and tryptophan were determined in plasma and blood. Paroxetine treatment reduced platelet 5-HT to 17% of baseline after 4 weeks of treatment. Responder patients had a pretreatment platelet 5-HT concentration 22% lower than nonresponders (p < .035). Admission HAMD scores, plasma paroxetine concentration, or platelet 5-HT concentration at endpoint did not differ between responders and nonresponders. Yet, the response rate was 11% in patients with high pretreatment platelet 5-HT (> 900 ng/10(9) platelets) and 50% in those below that value (p < .004). These findings support that depressed patients with a high pretreatment platelet 5-HT concentration have a poor therapeutic outcome after treatment with a standard paroxetine dose. These differences may be related to the existence of molecular differences in the 5-HT transporter.

  1. Propagation capacity of bacterial contaminants in platelet concentrates using a luciferase reporter system.

    PubMed

    Bello-López, Juan Manuel; Ibáñez-Cervantes, Gabriela; Fernández-Sánchez, Verónica; Arroyo-Pérez, José Antonio; Rojo-Medina, Julieta

    2015-06-01

    Currently the use of molecular tools and techniques of Genetic Engineering in the study of microbial behavior in blood components has replaced the employment of classical methods of microbiology. This work focuses on the use of a novel lux reporter system for monitoring the contaminating propagation capacity of bacteria present in platelet concentrates under standard storage conditions in the blood bank. A miniTn5 promotor probe carrying the lux operon from Photorhabdus luminiscens (pUTminiTn5luxCDABEKm2) was used to construct four bacterial bioluminescent mutants: Escherichia coli, Salmonella typhi, Proteus mirabilis and Pseudomonas aeruginosa. Luminescent mutants were used for contamination tests with 20 CFU in platelet concentrates bags and were stored under standard storage conditions in the blood bank (100 rpm at 22 °C). The measurements of luminous activity and optical density were used to monitor bacterial proliferation during 7 days (168 h). During the exponential growth phase (log) of bacterial strains, a lineal correlation between luminous activity vs biomass was observed (R(2) = 0.985, 0.976, 0.981) for E. coli::Tn5luxCDABEKm2, P. mirabilis::Tn5luxCDABEKm2 and P. auriginosa::Tn5luxCDABEKm2, respectively. The above indicates that metabolic activity (production of ATP) is directly related to biomass in this phase of microbial growth. While conducting experiments, the inability to propagate S. typhi::Tn5luxCDABEKm2 was detected. We can speculate that platelet concentrates contain specific components that prevent the propagation of S. typhi. The use of luxCDABE system for the quantification of luminous activity is a rapid and sensitive alternative to study the propagation and auto-sterilization of bacterial contaminants in platelet concentrates. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Applications of real-time PCR in the screening of platelet concentrates for bacterial contamination.

    PubMed

    Mohammadi, Tamimount; Savelkoul, Paul H M; Pietersz, Ruby N I; Reesink, Henk W

    2006-11-01

    Although there have been major improvements over the past few decades in detection methods for blood-borne infectious agents, platelet concentrates are still responsible for most cases of transfusion-transmitted bacterial infections. To date, real-time PCR is an indispensable tool in diagnostic laboratories to detect pathogens in a variety of biological samples. In this article, the applications of this powerful technique in the screening of platelet concentrates for bacterial contamination are discussed. Next to pathogen-specific (real-time) PCR assays, particular attention is directed to the recently developed 16S rDNA real-time PCR. This assay has been proven as a convenient way to detect bacterial contamination of platelet concentrates. The assay is sensitive and enables rapid detection of low initial numbers of bacteria in platelet concentrates. The short turnaround time of this assay allows high-throughput screening and reduction of the risk of transfusion of bacterially contaminated units. As with every method, real-time PCR has its advantages and disadvantages. These and especially limitations inherent to generation of false-positive or -negative results are emphasized. The universal nature of detection of the assay may be suitable for generalized bacterial screening of other blood components, such as red blood cells and plasma. Therefore, it is necessary to adapt and optimize detection in red blood cells and plasma with real-time PCR. Further sophistication, miniaturization and standardization of extraction and amplification methods should improve the total performance and robustness of the assay. Hence, real-time PCR is an attractive method in development as a more rapid screening test than currently used culture methods to detect bacterial contamination in blood components.

  3. Effect of platelet concentrate on quality of life after periradicular surgery: a randomized clinical study.

    PubMed

    Del Fabbro, Massimo; Ceresoli, Valentina; Lolato, Alessandra; Taschieri, Silvio

    2012-06-01

    Control of postoperative discomfort might enhance the patient's quality of life and treatment acceptance. The aim of the present randomized single-blind study was to evaluate whether the use of platelet concentrate during endodontic surgery might have a favorable impact on pain and other factors related to patient's quality of life during the first week after surgery. Eighteen patients with periapical lesion were treated with modern endodontic surgical procedure (control group). In another 18 patients, in adjunct to surgical procedure, platelet concentrate was applied on the root end in liquid form, within the bone defect in clot form, and over the suture in liquid form (test group). All patients completed a questionnaire for evaluation of main symptoms and daily activities during the first week after surgery. The outcomes of the questionnaires of the 2 groups were statistically compared. The test group showed significantly less pain and swelling, fewer analgesics taken, and improved functional activities as compared with the control group. The adjunct of platelet concentrate to the endodontic surgical procedure produced significant beneficial effect to patients' quality of life during the early postoperative stage. Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  4. Ultrastructural characteristics of fibrin clots from canine and feline platelet concentrates activated with calcium gluconate or calcium gluconate plus batroxobin

    PubMed Central

    2013-01-01

    Background The aim of this study was to use transmission electron microscopy to describe the ultrastructural characteristics of clots obtained from canine and feline platelet concentrates (PC) that had been activated with calcium gluconate (CG) or CG plus batroxobin (CGB). Platelets from fibrin clots were classified according their morphological changes. The area of the intercellular space (μm2), the area of the fibrin fibers (μm2), and the width of the fibrin fibers (μm) were determined for the dog clots. The platelet area (μm2), the area of fibrin fibers (μm2), the ratio of the minor and major axes of platelets, the ratio of the major and minor axes of platelets, and the number of α-granules found within platelets were measured for the cat clots. Results Cat platelets displayed full activation. Dog platelets displayed lysis with loss of normal architecture. In both species, a statistically significant difference was found (P < 0.01) between the fibrin fiber measurements in the PC clots activated with CG and CGB. Conclusions The findings suggest that activation with CG caused platelet alpha granules to release their contents. In cats, fibrin production was greater when the PC was activated with CG. In dogs, activation with CG produced thick fibrin fibers. PMID:23587176

  5. Serratia marcescens strains implicated in adverse transfusion reactions form biofilms in platelet concentrates and demonstrate reduced detection by automated culture.

    PubMed

    Greco-Stewart, V S; Brown, E E; Parr, C; Kalab, M; Jacobs, M R; Yomtovian, R A; Ramírez-Arcos, S M

    2012-04-01

    Serratia marcescens is a gram-negative bacterium that has been implicated in adverse transfusion reactions associated with contaminated platelet concentrates. The aim of this study was to investigate whether the ability of S. marcescens to form surface-attached aggregates (biofilms) could account for contaminated platelet units being missed during screening by the BacT/ALERT automated culture system. Seven S. marcescens strains, including biofilm-positive and biofilm-negative control strains and five isolates recovered from contaminated platelet concentrates, were grown in enriched Luria-Bertani medium and in platelets. Biofilm formation was examined by staining assay, dislodging experiments and scanning electron microscopy. Clinical strains were also analysed for their ability to evade detection by the BacT/ALERT system. All strains exhibited similar growth in medium and platelets. While only the biofilm-positive control strain formed biofilms in medium, this strain and three clinical isolates associated with transfusion reactions formed biofilms in platelet concentrates. The other two clinical strains, which had been captured during platelet screening by BacT/ALERT, failed to form biofilms in platelets. Biofilm-forming clinical isolates were approximately three times (P<0·05) more likely to be missed by BacT/ALERT screening than biofilm-negative strains. S. marcescens strains associated with transfusion reactions form biofilms under platelet storage conditions, and initial biofilm formation correlates with missed detection of contaminated platelet concentrates by the BacT/ALERT system. © 2011 The Author(s). Vox Sanguinis © 2011 International Society of Blood Transfusion.

  6. Comparison of various dimethylsulphoxide-containing solutions for cryopreservation of leucoreduced platelet concentrates.

    PubMed

    Dijkstra-Tiekstra, M J; de Korte, D; Pietersz, R N I; Reesink, H W; van der Meer, P F; Verhoeven, A J

    2003-11-01

    Leucoreduced platelet concentrates (LR-PCs) can be stored at 20-24 degrees C for 5-7 days. When LR-PCs are cryopreserved they can be stored for several years. For cryopreservation to become applicable in blood-bank practice, an off-the-shelf cryoprotectant is needed that can be added to the LR-PC in a sterile manner. For this, we varied the composition of the cryopreservation medium and studied various parameters of cryopreserved LR-PCs for up to 24 h after thawing at room temperature. LR-PCs in plasma or Composol were concentrated and divided into 2 units. To each unit, an equal part of 10% dimethylsulfoxide (DMSO) in plasma, Composol with or without 5% albumin, or GPO (pasteurized plasma-protein solution) was added. Freezing occurred at 1 degrees C/min and LR-PCs were placed in the vapour phase of nitrogen. LR-PCs were thawed at 37 degrees C and stored at room temperature. LR-PCs were tested for morphology, platelet recovery, swirling effect, and activation antigens at various time-points thereafter. LR-PCs in 100%, 65% and 50% plasma supplemented with Composol showed good morphology scores (>250), limited CD62P expression (<35%), low CD63 expression (<20%) and a swirling effect of about 2, at 24 h after thawing. At the same time-point, platelet recovery was >80% under all conditions and CD42b expression varied between 70 and 85%. Results of LR-PCs in 15% plasma and Composol, with or without plasma substitutes, were not acceptable at 24 h after thawing, i.e. the morphology score was <200 and the CD62P expression was >40%. A minimum of 50% plasma in the cryopreserved LR-PC is necessary to maintain an acceptable in vitro quality of platelets up to 24 h after thawing. Composol is a good candidate for using to prepare an off-the-shelf cryoprotectant.

  7. Blood Product Supply in Germany: The Impact of Apheresis and Pooled Platelet Concentrates

    PubMed Central

    Berger, Karin; Schopohl, Dorothee; Wittmann, Georg; Schramm, Wolfgang; Ostermann, Helmut; Rieger, Christina

    2016-01-01

    Background In Germany, about 60% of all produced platelet concentrates (PCs) are apheresis PCs (APCs). Ongoing discussions on APC reimbursement and costs might lead to a potential shift in pooled PC (PPC)/APC production. Objective of this analysis was to build a comprehensive model from the societal perspective to evaluate consequences associated with shifts in platelet supply and demand. Methods Literature search, desktop researches on platelet supply and demand. Model calculations, time horizon one year: model input from the Paul-Ehrlich-Institute, data 2013. Base case: 19.2% of annual whole blood donations (WBDs) were used for production of 38.5% PPCs, decay of 46,218 PCs (8.0%). Scenarios calculated: variation in PPC proportion of 10-100%. Results Base case: during PPC production 41,957-83,913 red blood cell concentrates (RBCCs) are estimated to be lost, which corresponds to 1-2% of annual RBCCs in Germany. Scenarios were calculated for a production of 60-100% PPCs: loss is estimated to be 1.5-5.0% of annual RBCCs (65,430-218,099), decay 54,189-69,022 PCs (9.4-12.0%). Conclusion Production of different blood components is interlinked and sensitive to unidimensional decisions. Increasing PPC proportion has negative impact on the RBCC production and on the antigen-matched APC donor pool. Completion of the model calculations to predict the optimal PPC/APC proportion would require evidence on the number of refractory patients, donor pool sizes, and incidences of diseases requiring platelet transfusions. PMID:27994524

  8. Clinically relevant HOCl concentrations reduce clot retraction rate via the inhibition of energy production in platelet mitochondria.

    PubMed

    Misztal, T; Rusak, T; Tomasiak, M

    2014-12-01

    Using porcine blood, we examined the impact of hypochlorite, product of activated inflammatory cells, on clot retraction (CR), an important step of hemostasis. We found that, in vitro, HOCl is able to reduce CR rate and enlarge final clot size in whole blood (t.c. 100 μM), platelet-rich plasma (PRP) threshold concentration (t.c. 50 μM), and an artificial system (washed platelets and fibrinogen) (t.c. 25 nM). Combination of low HOCl and peroxynitrite concentrations resulted in synergistic inhibition of CR by these stressors. Concentrations of HOCl completely inhibiting CR failed to affect the kinetics of coagulation measured in PRP and in platelet-free plasma. Concentrations of HOCl reducing CR rate in PRP augmented production of lactate, inhibited consumption of oxygen by platelets, and decreased total adenosine triphosphate (ATP) content in PRP-derived clots. In an artificial system, concentrations of HOCl resulting in inhibition of CR (25-100 nM) reduced mitochondrial transmembrane potential and did not affect actin polymerization in thrombin-stimulated platelets. These concentrations of HOCl failed to affect the adhesion of washed platelets to fibrinogen and to evoke sustained calcium signal, thus excluding stressor action on glycoprotein IIb/IIIa receptors. Exogenously added Mg-ATP almost completely recovered HOCl-mediated retardation of CR. Concentrations of HOCl higher than those affecting CR reduced thromboelastometric variables (maximum clot firmness and α angle). We conclude that low clinically relevant HOCl concentrations may evoke the inhibition of CR via the reduction of platelet contractility resulted from malfunction of platelet mitochondria. At the inflammatory conditions, CR may be the predominant HOCl target.

  9. In search of a consensus terminology in the field of platelet concentrates for surgical use: platelet-rich plasma (PRP), platelet-rich fibrin (PRF), fibrin gel polymerization and leukocytes.

    PubMed

    Dohan Ehrenfest, David M; Bielecki, Tomasz; Mishra, Allan; Borzini, Piero; Inchingolo, Francesco; Sammartino, Gilberto; Rasmusson, Lars; Evert, Peter A

    2012-06-01

    In the field of platelet concentrates for surgical use, most products are termed Platelet-Rich Plasma (PRP). Unfortunately, this term is very general and incomplete, leading to many confusions in the scientific database. In this article, a panel of experts discusses this issue and proposes an accurate and simple terminology system for platelet concentrates for surgical use. Four main categories of products can be easily defined, depending on their leukocyte content and fibrin architecture: Pure Platelet-Rich Plasma (P-PRP), such as cell separator PRP, Vivostat PRF or Anitua's PRGF; Leukocyteand Platelet-Rich Plasma (L-PRP), such as Curasan, Regen, Plateltex, SmartPReP, PCCS, Magellan, Angel or GPS PRP; Pure Plaletet-Rich Fibrin (P-PRF), such as Fibrinet; and Leukocyte- and Platelet-Rich Fibrin (L-PRF), such as Choukroun's PRF. P-PRP and L-PRP refer to the unactivated liquid form of these products, their activated versions being respectively named P-PRP gels and L-PRP gels. The purpose of this search for a terminology consensus is to plead for a more serious characterization of these products. Researchers have to be aware of the complex nature of these living biomaterials, in order to avoid misunderstandings and erroneous conclusions. Understanding the biomaterials or believing in the magic of growth factors ? From this choice depends the future of the field.

  10. Platelet-rich concentrates differentially release growth factors and induce cell migration in vitro.

    PubMed

    Schär, Michael O; Diaz-Romero, Jose; Kohl, Sandro; Zumstein, Matthias A; Nesic, Dobrila

    2015-05-01

    Platelet-rich concentrates are used as a source of growth factors to improve the healing process. The diverse preparation protocols and the gaps in knowledge of their biological properties complicate the interpretation of clinical results. In this study we aimed to (1) analyze the concentration and kinetics of growth factors released from leukocyte- and platelet-rich fibrin (L-PRF), leukocyte- and platelet-rich plasma (L-PRP), and natural blood clot during in vitro culture; (2) investigate the migration of mesenchymal stem cells (MSCs) and human umbilical vein endothelial cells (HUVECs) as a functional response to the factors released; and (3) uncover correlations between individual growth factors with the initial platelet/leukocyte counts or the induced cell migration. L-PRF, L-PRP, and natural blood clot prepared from 11 donors were cultured in vitro for 28 days and media supernatants collected after 8 hours and 1, 3, 7, 14, and 28 days. Released transforming growth factor β1 (TGF-β1), vascular endothelial growth factor (VEGF), insulin growth factor (IGF-1), platelet-derived growth factor AB (PDGF-AB), and interleukin-1β (IL-1β) were measured in the supernatants with enzyme-linked immunosorbent assay. Migration of MSC and HUVEC induced by the supernatants was evaluated in Boyden chambers. More TGF-ß1 was released (mean ± SD in pg/mL of blood) from L-PRF (37,796 ± 5492) compared with L-PRP (23,738 ± 6848; p < 0.001) and blood clot (3739 ± 4690; p < 0.001), whereas more VEGF and IL-1ß were released from blood clot (1933 ± 704 and 2053 ± 908, respectively) compared with both L-PRP (642 ± 208; p < 0.001 and 273 ± 386; p < 0.001, respectively) and L-PRF (852 ± 376; p < 0.001 and 65 ± 56, p < 0.001, respectively). No differences were observed in IGF-1 and PDGF-AB released from any of the concentrates. TGF-β1 release peaked at Day 7 in L-PRF and at 8 hours and Day 7 in L-PRP and 8 hours and Day 14 in blood clot. In all concentrates, main release of VEGF

  11. Reduced platelet concentration does not harm PRP effectiveness for ACL repair in a porcine in vivo model

    PubMed Central

    Mastrangelo, AN; Vavken, P; Fleming, BC; Harrison, SL; Murray, MM

    2011-01-01

    Enhanced primary repair of the ACL using a collagen scaffold loaded with platelets has been shown to improve the functional healing of suture repair in animal models. In this study, our objectives were to determine if lowering the platelet concentration would reduce the structural properties of the repaired ACL and increase postoperative knee laxity. Eight Yucatan minipigs underwent bilateral suture repair. In one knee, the repair was augmented with a collagen scaffold saturated with platelet-rich plasma containing five times the systemic baseline of platelets (5X) while the contralateral knee had a collagen scaffold saturated with platelet-rich plasma containing three times the systemic baseline of platelets (3X). After thirteen weeks of healing, knee joint laxity and the structural properties of the ACL were measured. The 3X platelet concentration resulted in a 24.1% decrease in cellular density of the repair tissue (p<0.05), but did not significantly decrease the structural properties [3Xvs 5X: 314 vs 298 N (p=0.596) and 65 vs 64 N/mm (p=0.532) for the yield load and linear stiffness, respectively]. The 3x platelet concentration also did not significantly change the mean anteroposterior knee laxity at 30° and 90° of flexion [5X vs. 3X: 3.5 vs. 5.1 mm (p=0.140), and 6.1 vs. 6.3 mm (p=0.764)] but did result in a lower AP laxity at 60° [5X vs. 3X: 8.6 vs. 7.3 mm (p=0.012)]. The decrease in platelet concentration from 5X to 3X to enhance suture repair of the ACL did not significantly harm the mechanical outcomes in this animal model. PMID:21337615

  12. Defining an appropriate leucoreduction strategy by serial assessment of cytokine levels in platelet concentrates prepared by different methods

    PubMed Central

    Kaur, Daljit; Sharma, Ratti Ram; Marwaha, Neelam

    2015-01-01

    Background and Objectives: Different methods of platelet concentrate preparations leave behind certain number of residual leukocytes, accounting for most of the febrile nonhemolytic transfusion reactions, especially in multitransfused patients. Various inflammatory cytokines, such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 are generated during storage and have been implicated for these adverse effects. We have studied the levels of these cytokines and their correlation with leucocyte contents in platelet concentrates prepared by three different methods. Study Design and Methods: Five pools of platelet rich plasma platelet concentrates (PRP-PC) and buffy-coat platelet concentrates (BC-PC) each were prepared and divided into two halves. One half of the pool was leucofiltered (LF), whereas the other half was stored as such. Ten apheresis units were also included in the study. All the platelet concentrates were assessed for leucocyte load and cytokine content (IL-1β, IL-6, and TNF-α) on different days of storage (0, 3, and 5) using Nageotte chamber and commercially available immunoassays respectively. Results: There was a statistically significant rise in cytokine levels (IL-1β, IL-6, and TNF-α) in nonleucofiltered (NLF) random donor platelet concentrates (RDPs) (PRP-PC and BC-PC) during storage (day 3 and 5) whereas LF RDP concentrates (PRP-PC and BC-PC) and apheresis platelet concentrates (AP-PC) did not show any significant rise in cytokine levels (on day 3 and 5) over the baseline values at day 0. Conclusion: This data suggests that although AP-PCs are superior to PRP-PC (NLF) and BC-PC (NLF) in terms of in vitro quality control parameters and cytokine generation during storage, BC-PC mode of platelet preparation followed by leucofiltration is the best method to store platelets and minimise the cytokine accumulation. This strategy is best suited for transfusion in multitransfused hematooncologic patients, who cannot afford single

  13. Storage of platelet concentrates after high-dose ultraviolet B irradiation

    SciTech Connect

    Snyder, E.L.; Beardsley, D.S.; Smith, B.R.; Horne, W.; Johnson, R.; Wooten, T.; Napychank, P.A.; Male, P.; Buchholz, D.H. )

    1991-07-01

    Ultraviolet B (UVB) irradiation of platelet concentrates (PCs) may prevent the development of posttransfusion HLA alloimmunization. As irradiation performed in a blood center or a hospital will probably be associated with a variable postirradiation delay before transfusion, the ability to store PCs after UVB irradiation becomes important. The effects have been studied of a UVB dose of 10,000 mJ per cm2, the dose used in our institution for UVB clinical trials, on PCs pooled and stored for up to 96 hours after irradiation. Results showed that after 96 hours of storage, though there were no changes in pH, platelet count, white cell count, percent discharge of lactate dehydrogenase, or {beta}-thromboglobulin, there were significant decreases in morphology score and osmotic recovery. These changes, however, were not evident after 24 hours of storage. Similarly, there was a 60% decrease in immunoreactive membrane glycoprotein (GP) Ib after 96 hours of storage, but these changes were not seen after 48 hours of storage. No changes were seen in levels of GPIIb/IIIa in either group during the 96 hours of storage. On computer-analyzed two-dimensional polyacrylamide gel electrophoresis, PCs irradiated at 10,000 mJ per cm2 and stored for 72 hours had changes in over 50 platelet proteins as compared to those proteins in nonirradiated age-matched control PCs. It can be concluded that UVB irradiation of PCs at 10,000 mJ per cm2 does not lead to significant platelet deterioration after short-term storage (24-48 hours) but is likely to be deleterious after long-term (72-96 hours) storage.

  14. Collection, storage, inspection and quality control of platelet concentrates obtained by apheresis: The situation in Spain.

    PubMed

    Castrillo, Azucena; Jimenez-Marco, Teresa; Arroyo, José L; Jurado, María L; Larrea, Luis; Maymo, Rosa M; Monge, Jorge; Muñoz, Carmen; Pajares, Ángel; Yáñez, Marta

    2017-06-01

    Diverse variables are involved in apheresis platelet collection, processing and storage. This survey shows how these are realized in Spain. An analysis of collected data was performed in a questionnaire completed by ten Transfusion Centers (TC) which perform between 50 and 520 apheresis procedures per month. This information comprises the procedures used to collect, inspect and store apheresis platelet concentrates (PC), and quality control data. Macroscopic inspection of PC is performed in all TC, especially during the first few hours post-collection and before distribution. The type of processor, duration of post-collection resting periods and temperature from the time of collection until distribution are similar in all TC. In 80% of TC, PC with small and scarce aggregates are distributed to transfusion services. The presence of clumps is influenced by type of processor, female donor, cold ambient temperature and collection of hyperconcentrated platelets, and is often recurrent in the same donor, although some TC have not found any influential variables. Overall, no objective inspection methods are followed, although there are exceptions. The degree of compliance with quality control parameters, such as the number of units studied, mean platelet yield, residual leukocyte counts and pH at expiry date, is acceptable in all TC. Compliance in terms of number of microbiological culture samples is variable. The usual practice in Spanish TC with respect to the collection, post-collection handling and storage of apheresis PC can be considered uniform, although some specific aspects of analyses should follow more objective methods. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. The 24-hour shelf-life of cytapheresis platelet concentrates stored in polyvinyl chloride containers should be extended only with caution.

    PubMed

    Strauss, R G; Snyder, E L; Eckermann, I; Stewart, L

    1987-01-01

    A recent publication suggested that the 24-hour allowable shelf-life of apheresis platelet concentrates collected by open-system techniques be extended to 48 hours because platelets collected in this fashion usually remain sterile for that length of time. The current studies, however, show that the quality of platelet concentrates deteriorates rapidly after storage for more than 24 hours in the relatively small-volume, polyvinyl chloride containers of currently marketed, open-system software, as evidenced by the falling pH, the disintegration of platelets, and the inability of platelets to recover from hypotonic shock. Platelets were markedly defective within 48 hours. Thus, it seems unwise to extend the shelf-life of such platelet concentrates beyond 24 hours solely because they are likely to remain sterile. Collection techniques and software must also be modified to ensure satisfactory platelet quality before the period of storage should be extended.

  16. Feasibility for EGRET detection of antimatter concentrations in the universe

    NASA Technical Reports Server (NTRS)

    Hartman, R. C.

    1990-01-01

    Although the Grand Unified Theories of elementary particle dynamics have to some extent reduced the aesthetic attraction of matter-antimatter symmetry in the Universe, the idea is still not ruled out. Although first introduced by Alfven (1965), most of the theoretical development related to gamma-ray astronomy was carried out by Stecker, who has proposed (Stecker, Morgan, and Bredekamp, 1971) matter-antimatter annihilation extending back to large redshifts as a possible explanation of the apparently extragalactic diffuse gamma radiation. Other candidate explanations were also proposed, such as superposition of extragalactic discrete sources. Clearly, the existence of significant amounts of antimatter in the universe would be of great cosmological importance; its detection, however, is not simple. Since the photon is its own antiparticle, it carries no signature identifying whether it originated in a matter or an antimatter process; even aggregates of photons (spectra) are expected to be identical from matter and antimatter processes. The only likely indicator of the presence of concentrations of antimatter is evidence of its annihilation with normal matter, assuming there is some region of contact or overlap. The EGRET (Energetic Gamma-Ray Experimental Telescope) on the Gamma Ray Observatory, with a substantial increase in sensitivity compared with earlier high energy gamma ray telescopes, may be able to address this issue. The feasibility of using EGRET in such a search for antimatter annihilation in the Universe is considered.

  17. Micro-concentration Lipopolysaccharide as a Novel Stimulator of Megakaryocytopoiesis that Synergizes with IL-6 for Platelet Production

    PubMed Central

    Wu, Di; Xie, Jun; Wang, Xuejun; Zou, Bingcheng; Yu, Yin; Jing, Tao; Zhang, Songmei; Zhang, Qing

    2015-01-01

    Lipopolysaccharide (LPS) induces platelet activation and enhances platelet sensitivity to aggregation, which might alter platelet counts. We found that serial doses of micro-concentration LPS significantly increased the platelet count in mice treated with kanamycin, along with increased expression of IL-6 compared with IL-3 and TPO in megakaryocytes obtained from the mouse bone morrow following LPS administration. Furthermore, LPS at lower levels ranging plus IL-6 effectively stimulated CFU-MK formation and increased CD41 expression and megakaryocyte polyploidization. Meanwhile, there was a sustained rise in the percentage of reticulated platelets in the whole blood in response to low-dosage LPS combined with IL-6. In vivo experiments also demonstrated that the administration of LPS combined with IL-6 substantially enhanced the number of circulating platelets in normal and thrombocytopenic mice. Notably, the optimal LPS concentration in combination with IL-6 might be a novel stimulator of TLR4 and IL-6R expression in Dami cell lines, which initially occurs through TLR4-IL-6R crosstalk and then involves the activation of NF-κB and phosphorylation of p38 MAPK. These data suggest a new paradigm for the regulation of megakaryocytopoiesis and platelet production via a synergistic effect of LPS and IL-6, which has the potential to be used for the design of new therapies. PMID:26330186

  18. Implications of anticoagulants and gender on cell counts and growth factor concentration in platelet-rich plasma and platelet-rich gel supernatants from rabbits.

    PubMed

    González, Juan C; López, Catalina; Carmona, Jorge U

    2016-01-01

    Our objectives were as follows: 1) to validate a protocol for producing rabbit platelet-rich plasma (PRP); 2) to determine the influence of two anticoagulants, sodium citrate and acid-citrate-dextrose solution A, and gender on cell count in PRP and growth factor concentration in pure platelet-rich gel supernatants; 3) to correlate the variables evaluated. Whole blood from 18 New Zealand rabbits (9 males and 9 females) was obtained with sodium citrate and acid-citrate-dextrose solution A for processing PRP fractions (A and B), which were evaluated for haematology. The PRP fractions were either activated with calcium gluconate or lysated with a detergent. The concentrations of transforming growth factor beta 1 and platelet-derived growth factor BB were assayed by ELISA. The sodium citrate PRP-B had significantly higher counts of platelets in comparison to PRP-A and whole blood obtained with the same anticoagulant and the homologous acid-citrate-dextrose solution A PRP fraction. The sodium citrate PRP-A had a significantly higher count of leukocytes compared to the homologous acid-citrate-dextrose solution A fraction. All the PRP fractions had a significant leuko-reduction when compared to whole blood. The sodium citrate PRP-A fraction from female rabbits had significantly lower platelet counts and significantly higher leukocyte counts than the same acid-citrate-dextrose solution A fraction. Growth factor concentration was not affected by the type of anticoagulant or gender. The type of anticoagulant and gender affected the cell counts in PRP, but they did not influence the growth factor concentration. More complete rabbit PRP studies should be performed before evaluating this type of substance in models of disease.

  19. The effect of platelet concentrates on graft maturation and graft-bone interface healing in ACL reconstruction in human patients

    PubMed Central

    Vavken, Patrick; Sadoghi, Patrick; Murray, Martha M

    2011-01-01

    Purpose To systematically review the current evidence for effects of platelet concentrates on (1) graft maturation and (2) graft-bone interface healing in ACL reconstruction in human, controlled trials, and for ensuing differences in clinical outcomes. Methods A systematic search of PubMed, CINAHL, EMBASE, CCTR and CDSR was performed for controlled trials of human ACL reconstruction with and without platelet concentrates. Data validity was assessed and data were collected on graft maturation, graft-bone interface healing and clinical outcome. Results Eight studies met the inclusion criteria. Seven studies reported on graft maturation with significantly better outcomes in the platelet groups in four, and large differences in means in two (underpowered) studies. Five studies report on tunnel healing, but four found no difference between groups. Three studies assessed clinical outcome but found no differences, regardless whether they had shown a benefical (1/3) or no effect (2/3) of platelets on graft and tunnel healing. Conclusion The current best evidence suggests that the addition of platelet concentrates to ACL reconstruction may have a beneficial effect on graft maturation and could improve it by 20–30% on average, but with substantial variability. The most likely mode of action is that treatment with platelets accelerates graft repopulation and remodeling, and this interpretation is supported by the existing data and biologically plausible. However, the current evidence also shows only a very limited influence of platelet concentrates on graft-bone interface healing and no significant difference in clinical outcomes. Clinical Relevance This systematic review collected evidence that the use of platelet concentrates may be a safe and inexpensive way to optimize graft maturation after ACL reconstruction, but there is no evidence for improved graft-bone interface healing or a significant difference in clinical outcomes. Level of Evidence Level IV, systematic

  20. Classification of platelet concentrates (Platelet-Rich Plasma-PRP, Platelet-Rich Fibrin-PRF) for topical and infiltrative use in orthopedic and sports medicine: current consensus, clinical implications and perspectives

    PubMed Central

    Dohan Ehrenfest, David M.; Andia, Isabel; Zumstein, Matthias A.; Zhang, Chang-Qing; Pinto, Nelson R.; Bielecki, Tomasz

    2014-01-01

    Summary Platelet concentrates for topical and infiltrative use – commonly termed Platetet-Rich Plasma (PRP) or Platelet-Rich Fibrin (PRF) – are used or tested as surgical adjuvants or regenerative medicine preparations in most medical fields, particularly in sports medicine and orthopaedic surgery. Even if these products offer interesting therapeutic perspectives, their clinical relevance is largely debated, as the literature on the topic is often confused and contradictory. The long history of these products was always associated with confusions, mostly related to the lack of consensual terminology, characterization and classification of the many products that were tested in the last 40 years. The current consensus is based on a simple classification system dividing the many products in 4 main families, based on their fibrin architecture and cell content: Pure Platelet-Rich Plasma (P-PRP), such as the PRGF-Endoret technique; Leukocyte- and Platelet-Rich Plasma (LPRP), such as Biomet GPS system; Pure Platelet-Rich Fibrin (P-PRF), such as Fibrinet; Leukocyte- and Platelet-Rich Fibrin (L-PRF), such as Intra-Spin L-PRF. The 4 main families of products present different biological signatures and mechanisms, and obvious differences for clinical applications. This classification serves as a basis for further investigations of the effects of these products. Perspectives of evolutions of this classification and terminology are also discussed, particularly concerning the impact of the cell content, preservation and activation on these products in sports medicine and orthopaedics. PMID:24932440

  1. Classification of platelet concentrates (Platelet-Rich Plasma-PRP, Platelet-Rich Fibrin-PRF) for topical and infiltrative use in orthopedic and sports medicine: current consensus, clinical implications and perspectives.

    PubMed

    Dohan Ehrenfest, David M; Andia, Isabel; Zumstein, Matthias A; Zhang, Chang-Qing; Pinto, Nelson R; Bielecki, Tomasz

    2014-01-01

    Platelet concentrates for topical and infiltrative use - commonly termed Platetet-Rich Plasma (PRP) or Platelet-Rich Fibrin (PRF) - are used or tested as surgical adjuvants or regenerative medicine preparations in most medical fields, particularly in sports medicine and orthopaedic surgery. Even if these products offer interesting therapeutic perspectives, their clinical relevance is largely debated, as the literature on the topic is often confused and contradictory. The long history of these products was always associated with confusions, mostly related to the lack of consensual terminology, characterization and classification of the many products that were tested in the last 40 years. The current consensus is based on a simple classification system dividing the many products in 4 main families, based on their fibrin architecture and cell content: Pure Platelet-Rich Plasma (P-PRP), such as the PRGF-Endoret technique; Leukocyte- and Platelet-Rich Plasma (LPRP), such as Biomet GPS system; Pure Platelet-Rich Fibrin (P-PRF), such as Fibrinet; Leukocyte- and Platelet-Rich Fibrin (L-PRF), such as Intra-Spin L-PRF. The 4 main families of products present different biological signatures and mechanisms, and obvious differences for clinical applications. This classification serves as a basis for further investigations of the effects of these products. Perspectives of evolutions of this classification and terminology are also discussed, particularly concerning the impact of the cell content, preservation and activation on these products in sports medicine and orthopaedics.

  2. Biologic Options for Articular Cartilage Wear (Platelet-Rich Plasma, Stem Cells, Bone Marrow Aspirate Concentrate).

    PubMed

    Kraeutler, Matthew J; Chahla, Jorge; LaPrade, Robert F; Pascual-Garrido, Cecilia

    2017-07-01

    Biological treatments for articular cartilage repair have gained in popularity in the past decade. Advantages of these therapies include minimal invasiveness, improved healing time, and faster recovery. Biological therapies for cartilage repair include platelet-rich plasma, bone marrow aspirate concentrate, and cell-based therapies. These methods have the added benefit of containing growth factors and/or stem cells that aid in recovery and regeneration. The purpose of this article is to review the current cartilage treatment options and the existing literature on outcomes, complications, and safety profile of these products for use in the knee and hip joints. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Micro-flow visualization of red blood cell-enhanced platelet concentration at sudden expansion.

    PubMed

    Zhao, Rui; Marhefka, Joie N; Shu, Fangjun; Hund, Samuel J; Kameneva, Marina V; Antaki, James F

    2008-07-01

    Microscopic steps and crevices are inevitable features within prosthetic blood-contacting devices. This study aimed to elucidate the thrombogenicity of the associated microscopic flow features by studying the transport of fluorescent platelet-sized particles in a suspension of red blood cells (RBCs) flowing through a 100 microm:200 microm sudden expansion. Micro-flow visualization revealed a strong influence of hematocrit upon the path of RBCs and spatial concentration of particles. At all flow rates studied (Re = 8.3-41.7) and hematocrit 20% and lower, RBC streamlines were found to detach from the microchannel wall creating an RBC-depleted zone inside the step that was much larger than the cells themselves. However, the observed distribution of particles was relatively homogeneous. By contrast, the RBC streamlines of samples with hematocrit equal to or greater than 30% more closely followed the contour of the microchannel, yet exhibited enhanced concentration of particles within the corner. The corresponding size of the cell depletion layer was comparable with the size of the cells. This study implies that local platelet concentration in blood within the physiological range of hematocrit can be elevated within the flow separation region of a sudden expansion and implicates the role of RBCs in causing this effect.

  4. Plasma concentration of platelet-derived microparticles is related to painful vaso-occlusive phenotype severity in sickle cell anemia.

    PubMed

    Nebor, Danitza; Bowers, Andre; Connes, Philippe; Hardy-Dessources, Marie-Dominique; Knight-Madden, Jennifer; Cumming, Vanessa; Reid, Marvin; Romana, Marc

    2014-01-01

    High plasma level of microparticles (MPs) deriving mainly from erythrocytes and platelets has been detected in sickle cell anemia (SCA) patients. Flow cytometry was used to determine the concentration of MPs in two groups of SCA patients exhibiting marked differences in painful vaso-occlusive crisis rates [a non-severe group (n = 17) and a severe group (n = 12)], and in a control group composed of healthy subjects (n = 20). A 3- to 4-fold increase of total MP plasma concentration was detected in SCA patients. Higher platelet-derived MPs concentration was detected in the severe SCA group while erythrocyte-derived MPs concentration was increased in the non-severe SCA patient group only. Our results suggest that plasma concentration of MPs shed by platelets is a biomarker of the vaso-occlusive phenotype-related severity.

  5. Alveolar Ridge Preservation After Tooth Extraction with DFDBA and Platelet Concentrates: A Radiographic Retrospective Study

    PubMed Central

    Baniasadi, Behrang

    2017-01-01

    Objectives: The purpose of this study was to evaluate vertical alveolar bone loss 3 months after tooth extraction when a technique of ridge preservation was applied using a particulate demineralized freeze-dried bone allograft 300 - 500 µm associated with platelet concentrates (platelet-rich-fibrin) in the form of gel and membranes. Material and Methods: A retrospective radiological clinical study was conducted on 56 patients for whom 95 extractions had been performed immediately followed by alveolar filling. Among the patients, 17 were smokers and 16 were provided with an immediate removable temporary prosthesis after extractions. Vertical bone loss was measured radiologically by panoramic X-ray before extractions and by a computed tomography scan 3 months after, at the level of mid-buccal bone wall, by two independent observers. For statistical analysis, Student’s t-test was performed to compare the mean bone loss between mono- and pluri-radicular teeth and to compare the mean bone loss between tobacco users versus non users and finally to compare the mean bone loss between individuals that had provisional removable prosthesis and those that had not. Results: Three months after tooth extraction, the mean of vertical loss of the mid-buccal bone wall was 0.72 (SD 0.71) mm (5.53% SD 5.19). No significant difference between bone loss at mono-radicular and pluri-radicular teeth (P = 0.982) was observed. There was no significant correlation between tobacco habits and bone loss (P = 0.2), nor between provisional removable prosthesis and bone loss (P = 0.786). Conclusion: These results indicate a good potential for the technique using Demineralized Freeze-Dried Bone Allograft 300 - 500 µm and platelet concentrates in alveolar bone preservation. PMID:28357003

  6. Clinical effectiveness of leucoreduced, pooled donor platelet concentrates, stored in plasma or additive solution with and without pathogen reduction.

    PubMed

    Kerkhoffs, Jean-Louis H; van Putten, Wim L J; Novotny, Viera M J; Te Boekhorst, Peter A W; Schipperus, Martin R; Zwaginga, Jaap Jan; van Pampus, Lizzy C M; de Greef, Georgine E; Luten, Marleen; Huijgens, Peter C; Brand, Anneke; van Rhenen, Dick J

    2010-07-01

    Pathogen reduction (PR) of platelet products increases costs and available clinical studies are equivocal with respect to clinical and haemostatic effectiveness. We conducted a multicentre, open-label, randomized, non-inferiority trial comparing the clinical effectiveness of buffy-coat derived leukoreduced platelet concentrates (PC) stored for up to 7 d in plasma with platelets stored in platelet additive solution III (PASIII) without and with treatment with amotosalen-HCl/ultraviolet-A (UVA) photochemical pathogen reduction (PR-PASIII). Primary endpoint of the study was 1-h corrected count increment (CCI). Secondary endpoints were 24-h CCI, bleeding, transfusion requirement of red cells and PC, platelet transfusion interval and adverse transfusion reactions. Compared to plasma-PC, in the intention to treat analysis of 278 evaluable patients the mean difference for the 1-h CCI of PR-PASIII-PC and PASIII-PC was -31% (P < 0.0001) and -9% (P = n.s.), respectively. Twenty-seven patients (32%) had bleeding events in the PR-PASIII arm, as compared to 19 (19%) in the plasma arm and 14 (15%) in the PASIII arm (P = 0.034). Despite the potential advantages of pathogen (and leucocyte) inactivation of amotosalen-HCl/UVA-treated platelet products, their clinical efficacy is inferior to platelets stored in plasma, warranting a critical reappraisal of employing this technique for clinical use.

  7. Use of 8-methoxypsoralen and long-wavelength ultraviolet radiation for decontamination of platelet concentrates

    NASA Astrophysics Data System (ADS)

    Corash, Laurence; Lin, Lily; Wiesehahn, Gary; Cimino, George

    1992-06-01

    Transmission of viral diseases through blood products remains a problem in transfusion medicine. A number of methods have been developed to inactivate viral pathogens in plasma and plasma fractions, including: dry heating, wet heating, solvent-detergent treatment, and immunoaffinity purification. While some of these methods successfully inactivate pathogenic viruses, inactivation may be incomplete or result in damage to labile plasma proteins and cells. We have developed a photochemical decontamination system (PCD) for platelet concentrates (PC) utilizing treatment with long wavelength ultraviolet radiation (UVA, 320 - 400 nm) and 8-methoxypsoralen (8-MOP). This system is capable of inactivating 25 - 30 logs/hr of bacteria E. coli or S. aureus, 6 logs/hr of bacteriophage fd, 0.9 log/hr of bacteriophage R17 and 1.1 logs/hr of feline leukemia virus (FeLV) in PC. Immediately following 6 hrs of PCD treatment, platelet integrity and function of PCD treated and control PC were equivalent. After overnight storage PCD treated and control PC platelet properties were equal, but there was a slight reduction in TXB-2 production of PCD treated PC compared to controls. Following PCD treatment, PC were stored for 48 to 96 hrs. Platelet counts, morphology scores, extracellular LDH levels, aggregation response, dense body (db) content, and alpha granule ((alpha) g) content of PCD treated and control PC were comparable. We assessed the ability of the PCD technique to inactivate intracellular and extracellular virus, quantified the degree of DNA adduct formation in contaminating lymphocytes, and measured the inhibition of polymerase chain reaction (PCR) mediated amplification of intracellular DNA. High titers of cell-free murine cytomegalovirus added to human platelet concentrates (final concentration 106) were inactivated by PCD within 30 min. Cat renal fibroblasts infected at high levels with feline rhinotracheitis virus (FeRTV) were seeded into PC followed by PCD treatment with

  8. Which platelet function test best reflects the in vivo plasma concentrations of ticagrelor and its active metabolite? The HARMONIC study.

    PubMed

    Koziński, Marek; Ostrowska, Małgorzata; Adamski, Piotr; Sikora, Joanna; Sikora, Adam; Karczmarska-Wódzka, Aleksandra; Marszałł, Michał Piotr; Boinska, Joanna; Laskowska, Ewa; Obońska, Ewa; Fabiszak, Tomasz; Kubica, Jacek

    2016-11-30

    Aim of this study was assessment of the relationship between concentrations of ticagrelor and its active metabolite (AR-C124910XX) and results of selected platelet function tests. In a single-centre, cohort study, patients with myocardial infarction underwent blood sampling following a 180 mg ticagrelor loading dose intake (predose, 1, 2, 3, 4, 6, 12, 24 hours postdose) to perform pharmacokinetic and pharmacodynamic assessments. Platelet reactivity was evaluated using the VASP-assay, the VerifyNow device and the Multiplate analyzer. Analysis of 36 patients revealed high negative correlations between ticagrelor concentrations and platelet reactivity evaluated with all three platelet function tests (the VASP-assay: RS=-0.722; p<0.0001; the VerifyNow device: RS=-0.715; p<0.0001; the Multiplate analyzer: RS=-0.722; p<0.0001), with no significant differences between correlation coefficients. Similar results were found for AR-C124910XX. Platelet reactivity values assessed with all three methods generally correlated well with each other; however, a significantly higher correlation (p<0.02) was demonstrated between the VerifyNow and Multiplate tests (RS=0.707; p<0.0001) than in other assay combinations (the VASP-assay and the VerifyNow device: RS=0.595; p<0.0001; the VASP-assay and the Multiplate analyzer: RS=0.588; p<0.0001). With respect to the recognition of high platelet reactivity, we found higher measurement concordance between the VerifyNow and Multiplate tests compared with other assay combinations, while for low platelet reactivity, only results of the VerifyNow and Multiplate assay were related to each other. Platelet reactivity measurements performed with the VASP, VerifyNow and Multiplate tests show comparably strong negative correlations with ticagrelor and AR-C124910XX concentrations.

  9. Methodological Quality Assessment of Systematic Reviews on Autologous Platelet Concentrates for the Treatment of Periodontal Defects.

    PubMed

    Del Fabbro, Massimo; Lolato, Alessandra; Panda, Saurav; Corbella, Stefano; Satpathy, Anurag; Das, Abhaya Chandra; Kumar, Manoj; Taschieri, Silvio

    2017-09-01

    Evaluation of the methodological quality of systematic reviews (SRs) on the effectiveness of autologous platelet concentrates as an adjunct to regenerative procedures for the treatment of periodontal defects. After a literature screening, eligible SRs were qualitatively assessed using 2 validated instruments: A Measurement Tool to Assess systematic Reviews checklist and Overview Quality Assessment Questionnaire. The characteristics and findings of SRs were also reported. Ten SRs fulfilled the inclusion criteria and were evaluated. With A MeaSurement Tool to Assess systematic Reviews tool, SRs displayed a generally satisfying quality. Six SRs satisfied ≥8 items of 11 (high-quality score), and 4 were classified of medium quality (score 4-7). Using Overview Quality Assessment Questionnaire instrument, more than half SRs (N = 6) satisfied ≥7 items of 9, resulting to be of high quality; 3 were classified as medium quality (4-6 criteria met); and only 1 of low quality (3 items satisfied). A significant correlation between the results of the 2 questionnaires was found (Spearman's r = 0.915, P = .0005). SRs considered had an overall high methodological quality. However, some areas were not systematically addressed, like a thorough research strategy or publication bias assessment. Standard guidelines for designing, performing, and reporting SRs should always be followed. The use of platelet concentrates as an adjunct to periodontal surgery procedures may have beneficial effects for the treatment of periodontal defects. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Effect of solute concentration on intracellular water volume and hydraulic conductivity of human blood platelets.

    PubMed Central

    Armitage, W J

    1986-01-01

    The intracellular water volume of human blood platelets was determined using tritiated water. The cells responded as osmometers over an observed range of solute concentration from 0.292 to 2.180 osmol kg-1. Only 87% of intracellular water was apparently osmotically active (i.e. Ponder's R was 0.87). Changes in cell volume induced by small step changes in external osmolality were followed photometrically and the time constant for the exponential approach of cell volume to its new equilibrium value was determined. Hydraulic conductivity (LP) was calculated from the time constant and was 1.41 X 10(-6) cm atm-1 s-1 under isotonic conditions at 37 degrees C. LP was inversely dependent on extracellular solute concentration, but it was independent of the direction of movement of water across the plasma membrane. PMID:3746695

  11. Use of 8-methoxypsoralen and ultraviolet-A pretreated platelet concentrates to prevent alloimmunization against class I major histocompatibility antigens

    SciTech Connect

    Grana, N.H.; Kao, K.J. )

    1991-06-01

    The use of 8-methoxypsoralen (8-MOP) and UV-A irradiation to inactivate contaminating donor leukocytes in platelet concentrates and to prevent primary alloimmunization against donor class I major histocompatibility (MHC) antigens in mice was investigated. CBA/CaH-T6J mice with the H2k haplotype and BALB/cByJ mice with the H2d haplotype were used as donors and recipients, respectively. The mixed leukocyte reaction between these two strains of mice showed that treatment of spleen cells with 500 ng/mL 8-MOP and 5J/cm2 UV-A inhibited 99% of responder and 92% of stimulator function. There was no measurable loss of platelet aggregating activity after the treatment. After two weekly transfusions of platelets without any treatment, 93% of control mice (n = 15) developed anti-H2k antibody. In contrast, only 33% of mice (n = 15) receiving platelets treated with 8-MOP and UV-A became alloimmunized. After six weekly platelet transfusions, all mice became alloimmunized. Nevertheless, the mean titers of anti-H2k antibody in sera of the treated groups were significantly lower than the control groups. One hour posttransfusion recoveries of 51Cr-labeled donor platelets were also higher in mice transfused with the treated platelets. Thus, the pretreatment of platelet concentrates with 8-MOP and UV-A irradiation effectively reduced the alloantigenicity of class I MHC molecules. The implication of this finding in relation to the mechanism by which donor leukocytes allosensitize recipients is discussed.

  12. 21 CFR 864.9575 - Environmental chamber for storage of platelet concentrate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... to hold platelet-rich plasma within a preselected temperature range. (b) Classification. Class II... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Environmental chamber for storage of platelet... Establishments That Manufacture Blood and Blood Products § 864.9575 Environmental chamber for storage of platelet...

  13. In vivo evaluation of titanium-prepared platelet-rich fibrin (T-PRF): a new platelet concentrate.

    PubMed

    Tunalı, Mustafa; Özdemir, Hakan; Küçükodacı, Zafer; Akman, Serhan; Fıratlı, Erhan

    2013-07-01

    We have developed a new, titanium-prepared, platelet-rich fibrin (T-PRF) together with the protocol for forming it, which is based on the hypothesis that titanium tubes may be more effective at activating platelets than the glass tubes used by Chouckroun in his platelet-rich fibrin (PRF) method. The aim of this study was to find a suitable animal model in which to evaluate the method and to investigate the efficacy of T-PRF for wound healing. Blood samples from 6 rabbits were used to confirm the protocol for formation of T-PRF. We evaluated T-PRF or T-PRF-like clots morphologically using scanning electron microscopy (EM). Blood samples from 5 rabbits were used to develop an experiment in which to evaluate the effects of T-PRF on wound healing. The mucoperiosteal flaps were filled with autologous T-PRF membranes from the vestibule in the anterior mandibular regions. Samples collected from the surgical sites were stained with haematoxylin and eosin. We found a mature fibrin network in T-PRF clots that had been centrifuged for 15 min at 3500 rpm and, 15 days after placement of the membrane, we found newly-forming connective tissue and islets of bony tissue in the T-PRF membrane. These results show that T-PRF could induce the formation of new bone with new connective tissue in a rabbit model of wound healing within 30 days of treatment. Published by Elsevier Ltd.

  14. Storage of washed platelets in BRS-A platelet additive solutions based on two types of clinically available bicarbonated Ringer's solutions with different electrolyte concentrations.

    PubMed

    Oikawa, Shinji; Taguchi, Takeshi; Endo, Kimika; Hoshi, Takahiro; Kawashima, Wataru; Horibe, Yasuhito; Urano, Shinichi; Suzuki, Ko; Minegishi, Masayoshi; Itoh, Takashi; Shimizu, Hiroshi

    2015-10-01

    In Japan, no platelet (PLT) additive solutions (PASs) are officially approved for clinical use although blood centers often receive requests for washed PLTs to reduce adverse reactions. Recently, we developed a novel PAS called BRS-A based on clinically available bicarbonated Ringer's solution (BRS), Bicanate and acid-citrate-dextrose formula A (ACD-A), which has been shown to maintain the in vitro properties of PLTs in the condition of <5% residual plasma during 7-day storage. The aim of this study was to evaluate whether another clinically available BRS, Bicarbon with different electrolyte concentrations can be used as a PAS. Two types of BRS-As were prepared by adding 25 mL of ACD-A to 500 mL of Bicanate or Bicarbon BRSs. Bicanate-based BRS-A and Bicarbon-based BRS-A contain 0.9 or 0.5 mmol/L of magnesium chloride, 95.2 or 100.1 mmol/L of sodium chloride, 4.2 or 5.1 mmol/L of trisodium citrate, and 26.6 or 23.8 mmol/L of sodium bicarbonate, respectively; the other components were identical. Apheresis PLTs stored in these solutions with less than 5% plasma for 7-day storage were compared with regard to their in vitro properties. The pH levels of all units were above 7 throughout storage. The mean PLT volume, hypotonic shock response, glucose consumption, lactate production, swirling, and CD62P and CD42b expression were similar during 7-day storage. The bicarbonate levels in Bicarbon-based BRS-A were lower than those in Bicanate-based BRS-A. Differences in concentrations of electrolytes such as magnesium, sodium, citrate, and bicarbonate salts in BRS-A do not affect the in vitro properties of PLTs during 7-day storage. These results indicate that the use of another type of BRS-A based on Bicarbon as a PAS is feasible. Thus, BRS-A can be used in hospitals that do not stock Bicanate but have Bicarbon. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Association of plasma concentrations of salicylic acid and high on ASA platelet reactivity in type 2 diabetes patients.

    PubMed

    Postula, Marek; Janicki, Piotr K; Rosiak, Marek; Przybylkowski, Adam; Kaplon-Cieslicka, Agnieszka; Grygorowicz, Tomasz; Trzepla, Ewa; Filipiak, Krzysztof J; Czlonkowski, Andrzej; Opolski, Grzegorz

    2013-01-01

    The objective of this study was to investigate the association between plasma concentrations of salicylic acid (SA) and other minor acetylsalicylic acid (ASA) metabolites and high on ASA platelet reactivity assessed with different methods in type 2 diabetic patients (T2DM). Study cohort consisted of 293 T2DM patients on chronic ASA therapy. Platelet function inhibition was analyzed using measurements of serum thromboxane B2 (S-TxB2), VerifyNow Aspirin and Platelet Function Analyzer (PFA)-100 assays. The concentration of ASA metabolites in plasma was measured with a high-performance liquid chromatography (HPLC). In logistic regression analysis both ASA dose/kg of body weight and plasma SA concentration were found to be predictive of S-TxB2 concentrations above 0.72 ng/mL cut-off point (OR 16.9, 95% CI 2.29-125.8, p = 0.006 and OR 5.34, 95% CI 2.67-10.68, p < 0.001, respectively). When using the VerifyNow Aspirin Assay, the concentrations of SA were significantly lower (p = 0.007) in the group with high on ASA platelet reactivity when compared with the group with normal on ASA platelet reactivity. In logistic regression analysis plasma SA concentration was found to be predictive of VerifyNow Aspirin Reaction Units (ARU) ≥ 550 (OR 3.86, 95% CI 1.86-8.00, p < 0.001). Our study suggests that disturbances of pharmacokinetic mechanisms might contribute to lower plasma SA levels, and subsequently incomplete inhibition of thromboxane A2 synthesis as measured with S-TxB2 concentrations and increased platelet reactivity measured with VerifyNow in T2DM patients.

  16. Autologous Platelet Concentrates for Pulp and Dentin Regeneration: A Literature Review of Animal Studies.

    PubMed

    Del Fabbro, Massimo; Lolato, Alessandra; Bucchi, Cristina; Taschieri, Silvio; Weinstein, Roberto L

    2016-02-01

    The purpose of this study was to evaluate the effectiveness of autologous platelet concentrates (APCs) in promoting pulp and dentin regeneration in animal models. An electronic search was performed on MEDLINE, Embase, Scopus, SciELO, LILACS, and CENTRAL. Animal studies using APC as a root filling material after pulpectomy in mature or immature teeth were included. Articles underwent risk of bias assessment. Histologic evaluation of intracanal neoformed tissue was the primary outcome; root development, root wall thickening, apical closure, and periapical healing in apical periodontitis were the secondary outcomes. Seven articles were included. Platelet-rich plasma (PRP) was used as root filling material during regenerative procedures in the experimental group in either mature or immature teeth. After revascularization with PRP alone or in conjunction with stem cells of a different source, the histologic analyses revealed that, in addition to an odontoblastic cell layer or dentinlike structure, the neoformed intracanal tissues were mainly cementumlike, bonelike, and connective tissues. True regeneration of necrotic pulp may not be achieved with current techniques using PRP, all of which stimulated tissue repair. Benefits of PRP adjunct for pulp tissue regeneration in preclinical studies remain unclear. Further studies with standardized protocols are necessary to assess the actual contribution of PRP in endodontic regenerative therapies. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  17. Feasibility of implementing an automated culture system for bacteria screening in platelets in the blood bank routine.

    PubMed

    Castro, E; Bueno, J L; Barea, L; González, R

    2005-06-01

    Bacterial contamination of blood components is the principal infectious complication linked to transfusion. The aim of the study was to evaluate the applicability of an automated culture system for platelets. 10 141 platelet concentrates were cultured individually and in pools of five on storage days 1 and 7 using Bact/Alert system aerobic bottles. A modified collection bag was used for improved sampling. Five-millilitre samples were cultured at 37 degrees C for 7 days. Only those samples where the same bacteria were identified in reculture were considered true positives (TP). Homogeneity of proportions was tested by Fisher's exact test. The rate of TP was 30 per 100 000 (95% CI, 6.1-86.4) sampling on day 1; 33 per 100 000 (95% CI, 7-96) on day 7; and 40 per 100 000 (95% CI, 1.28-122.4) if the screening was based on taking both samples (day 1 and 7). Only one TP was detected in the pool testing. The time for detection among TPs on day 1 ranged between 30 and 134 h. The system is not considered practical for use as a routine screening method, as the time for detection is too long. Pool testing is insensitive. Faster screening methods or pathogen-inactivation systems are needed.

  18. The kinetics of changes in intracellular calcium concentration in fura-2-loaded human platelets.

    PubMed

    Sage, S O; Rink, T J

    1987-12-05

    We have investigated the sub-second kinetics of changes in cytosolic free calcium, [Ca2+]i, in fura-2-loaded human platelets by stopped-flow fluorimetry. Thrombin, vasopressin, platelet-activating factor, and the thromboxane A2 analogue U46619 all evoked a rise in [Ca2+]i which was delayed in onset by 200-400 ms in the presence of 1 mM external Ca2+. The responses to these agonists in media containing 1 mM EGTA or 1 mM Ni2+, to prevent Ca2+ influx, were delayed by an additional 60-100 ms. These results indicate that agonist-evoked Ca2+ influx precedes the release of Ca2+ from internal stores. The delays in onset of both responses are sufficient for one or more biochemical steps to lie between ligand-receptor binding and Ca2+ flux generation. ADP responses in media containing EGTA or Ni2+ were similar to those evoked by other agonists, but the response in the presence of external Ca2+ was markedly shorter, occurring without measurable delay at optimal ligand concentration. Analysis of this response showed some delay in ADP-evoked influx at lower concentrations, but this delay was markedly less than that observed with thrombin at doses giving the same elevation in [Ca2+]i. These results suggest that ADP evokes influx using a different transduction system, more closely coupled to the Ca2+ entry system than that used by other agonists. Differences between thrombin- and ADP-evoked influx were further demonstrated by the inhibitory actions of cAMP, which reduced and substantially increased the delay in onset of thrombin-evoked influx but did not measurably delay the influx evoked by an optimal concentration of ADP.

  19. Intramammary administration of platelet concentrate as an unconventional therapy in bovine mastitis: first clinical application.

    PubMed

    Lange-Consiglio, A; Spelta, C; Garlappi, R; Luini, M; Cremonesi, F

    2014-10-01

    Bovine udder infections induce a variety of changes in gene expression of different growth factors that may suggest their possible role in glandular tissue protection or repair processes. Growth factors and also chemokines and cytokines may act synergistically to increase the infiltration of neutrophils and macrophages to promote angiogenesis, fibroplasia, matrix deposition, and, ultimately, re-epithelialization. Considering the vast applications, typically in human medicine, of platelet concentrate (PC) and its ease of preparation, the aim of our study was to evaluate an alternative therapy to stimulate the regeneration of glandular tissue, administering a concentration in excess of the growth factors contained in the PC. In each one of the 3 farms examined in the trial, PC was prepared from donor cows in good health, free from infections, and with no records of medications administered during the previous 2 mo. The platelet produced in one farm was used only for treating the cows of the same farm in a heterologous way. A total of 229 mastitic quarters were divided in 3 groups: antibiotic group (treated with intramammary antibiotic), antibiotic and PC group (treated intramammarily with antibiotics in association with PC), and PC group (treated with intramammary PC alone). The diagnosis of mastitis was based on somatic cell count and bacteriological evaluation of the milk from the affected quarter. Platelet concentrate, alone or in association with antibiotic, was used for 3 consecutive days as an unconventional therapy in bovine acute and chronic mastitis. Our data show that the associated action of antibiotic and PC performed significantly better than the antibiotic alone, either for the recovery of the affected mammary quarters or for somatic cell count reduction. In the same way, the association antibiotic plus PC showed significantly fewer relapses compared with the antibiotic alone, either for acute or chronic mastitis. The treatment with only PC did not show

  20. Comparison between the effects of platelet-rich plasma and bone marrow concentrate on defect consolidation in the rabbit tibia

    PubMed Central

    Batista, Marco Antonio; Leivas, Tomaz Puga; Rodrigues, Consuelo Junqueira; Arenas, Géssica Cantadori Funes; Belitardo, Donizeti Rodrigues; Guarniero, Roberto

    2011-01-01

    OBJECTIVE: To perform a comparative analysis of the effects of platelet-rich plasma and centrifuged bone marrow aspirate on the induction of bone healing in rabbits. METHOD: Twenty adult, male New Zealand rabbits were randomly separated into two equal groups, and surgery was performed to create a bone defect (a cortical orifice 3.3 mm in diameter) in the proximal metaphysis of each rabbit's right tibia. In the first group, platelet-rich plasma was implanted in combination with β-tricalcium phosphate (platelet-rich plasma group), and in the second group, centrifuged bone marrow in combination with β-tricalcium phosphate (centrifuged bone marrow group) was implanted. After a period of four weeks, the animals were euthanized, and the tibias were evaluated using digital radiography, computed tomography, and histomorphometry. RESULTS: Seven samples from each group were evaluated. The radiographic evaluation confirmed the absence of fractures in the postoperative limb and identified whether bone consolidation had occurred. The tomographic evaluation revealed a greater amount of consolidation and the formation of a greater cortical bone thickness in the platelet-rich plasma group. The histomorphometry revealed a greater bone density in the platelet-rich plasma group compared with the centrifuged bone marrow group. CONCLUSION: After four weeks, the platelet-rich plasma promoted a greater amount of bone consolidation than the bone marrow aspirate concentrate. PMID:22012052

  1. Growth differentiation factor 11 (GDF11) - a promising anti-ageing factor - is highly concentrated in platelets.

    PubMed

    Bueno, J L; Ynigo, M; de Miguel, C; Gonzalo-Daganzo, R M; Richart, A; Vilches, C; Regidor, C; García-Marco, J A; Flores-Ballester, E; Cabrera, J R

    2016-11-01

    Recent research suggests that growth differentiation factor 11 (GDF11) could reverse age-related diseases and that its blood concentration decreases with age. This poses plasma from young donors as a therapeutic GDF11 source to treat age-related diseases. In addition, the tissue source of circulating GDF11 remains unknown. We analysed GDF11 levels in paired samples of serum, plasma and platelet lysate (PL) from 23 volunteers. Plasma and PL were collected by plateletpheresis. Here, we show that GDF11 is highly concentrated in platelets and that the circulating levels reported in previous studies could be biased as a result of serum sample manipulation.

  2. Comparison of the effect of calcium gluconate and batroxobin on the release of transforming growth factor beta 1 in canine platelet concentrates

    PubMed Central

    2012-01-01

    Background The clinical use of autologous platelet concentrates (also known as platelet-rich plasma) on the field of regenerative therapy, in the last decade has been the subject of several studies especially in equine medicine and surgery. The objectives of this study was: 1) to describe and compare the cellular population in whole blood, lower fraction (A) and upper fraction (B) of platelet concentrates, 2) to measure and compare the transforming growth factor beta 1 (TGF-β1) concentration in plasma and both platelet concentrates after be activated with calcium gluconate or batroxobin plus calcium gluconate and, 3) to determine correlations between cell counts in platelet concentrates and concentrations of TGF-β1. Blood samples were taken from 16 dogs for complete blood count, plasma collection and platelet concentrates preparation. The platelet concentrates (PC) were arbitrarily divided into two fractions, specifically, PC-A (lower fraction) and PC-B (upper fraction). The Platelet concentrates were analyzed by hemogram. After activated with calcium gluconate or batroxobin plus calcium gluconate, TGF-β1 concentration was determined in supernatants of platelet concentrates and plasma. Results There were differences statistically significant (P < 0.05) for the platelet count and leukocyte count and TGF-β1 concentration between whole blood, plasma and both platelet concentrates. A significant correlation was found between the number of platelets in both platelet concentrates and TGF-β1 concentration. Platelet collection efficiency was 46.34% and 28.16% for PC-A and PC-B, respectively. TGF-β1 concentration efficiency for PC activated with calcium gluconate was 47.75% and 31.77%, for PC-A and PC-B, respectively. PC activated with batroxobin plus CG showed 46.87% and 32.24% for PC-A and PC-B, respectively. Conclusions The methodology used in this study allows the concentration of a number of platelets and TGF-β1 that might be acceptable for a biological

  3. Do ambulatory-use Platelet-Rich Plasma (PRP) concentrates present risks?

    PubMed

    Martinez-Gonzalez, J M; Cano-Sanchez, J; Gonzalo-Lafuente, J C; Campo-Trapero, J; Esparza-Gomez, G; Seoane, J

    2002-01-01

    Platelet-Rich Plasma (PRP) concentrates have been widely used in the past decade as a complement to tissue regeneration procedures. The authors who have clinically used PRP refer no risk of infection, disease transmission, or undesirable effects. Nevertheless, there have been reports on the over-expression of growth factors (GFs) and their receptors related to tumour and dysplastic tissues. This has led to evaluation of the possible coincidences between carcinogenesis and the mitogenic pathways employed by GFs. The present study provides a review of the literature on the possible effects of the therapeutic uses of GFs (including PRP) in relation to carcinogenesis, their influence upon tissues with epithelial dysplasia or oral carcinoma, and their relation to tumour growth and infiltration.

  4. Autologous keratinocyte suspension in platelet concentrate accelerates and enhances wound healing – a prospective randomized clinical trial on skin graft donor sites: platelet concentrate and keratinocytes on donor sites

    PubMed Central

    2013-01-01

    Background Wound healing involves complex mechanisms, which, if properly chaperoned, can enhance patient recovery. The abilities of platelets and keratinocytes may be harnessed in order to stimulate wound healing through the formation of platelet clots, the release of several growth factors and cytokines, and cell proliferation. The aim of the study was to test whether autologous keratinocyte suspensions in platelet concentrate would improve wound healing. The study was conducted at the Lausanne University Hospital, Switzerland in 45 patients, randomized to three different topical treatment groups: standard treatment serving as control, autologous platelet concentrate (PC) and keratinocytes suspended in autologous platelet concentrate (PC + K). Split thickness skin graft donor sites were chosen on the anterolateral thighs of patients undergoing plastic surgery for a variety of defects. Wound healing was assessed by the duration and quality of the healing process. Pain intensity was evaluated at day five. Results Healing time was reduced from 13.9 ± 0.5 days (mean ± SEM) in the control group to 7.2 ± 0.2 days in the PC group (P < 0.01). An addition of keratinocytes in suspension further reduced the healing time to 5.7 ± 0.2 days. Pain was reduced in both the PC and PC + K groups. Data showed a statistically detectable advantage of using PC + K over PC alone (P < 0.01). Conclusion The results demonstrate the positive contribution of autologous platelets combined with keratinocytes in stimulating wound healing and reducing pain. This strikingly simple approach could have a significant impact on patient care, especially critically burned victims for whom time is of the essence. Clinical trial registry information Protocol Record Identification Number: 132/03 Registry URL: http://www.clinicaltrials.gov PMID:23570605

  5. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part V: histologic evaluations of PRF effects on bone allograft maturation in sinus lift.

    PubMed

    Choukroun, Joseph; Diss, Antoine; Simonpieri, Alain; Girard, Marie-Odile; Schoeffler, Christian; Dohan, Steve L; Dohan, Anthony J J; Mouhyi, Jaafar; Dohan, David M

    2006-03-01

    Platelet-rich fibrin (PRF) belongs to a new generation of platelet concentrates, with simplified processing and without biochemical blood handling. The use of platelet gel to improve bone regeneration is a recent technique in implantology. However, the biologic properties and real effects of such products remain controversial. In this article, we therefore attempt to evaluate the potential of PRF in combination with freeze-dried bone allograft (FDBA) (Phoenix; TBF, France) to enhance bone regeneration in sinus floor elevation. Nine sinus floor augmentations were performed. In 6 sites, PRF was added to FDBA particles (test group), and in 3 sites FDBA without PRF was used (control group). Four months later for the test group and 8 months later for the control group, bone specimens were harvested from the augmented region during the implant insertion procedure. These specimens were treated for histologic analysis. Histologic evaluations reveal the presence of residual bone surrounded by newly formed bone and connective tissue. After 4 months of healing time, histologic maturation of the test group appears to be identical to that of the control group after a period of 8 months. Moreover, the quantities of newly formed bone were equivalent between the 2 protocols. Sinus floor augmentation with FDBA and PRF leads to a reduction of healing time prior to implant placement. From a histologic point of view, this healing time could be reduced to 4 months, but large-scale studies are still necessary to validate these first results.

  6. Optimization of platelet concentrate quality: application of proteomic technologies to donor management.

    PubMed

    Schubert, Peter; Culibrk, Brankica; Karwal, Simrath; Slichter, Sherrill J; Devine, Dana V

    2012-12-05

    Quality management of blood products is essential for blood banking. It is influenced by both processing and donor characteristics and assured by monitoring routine in vitro parameters to defined product specifications. However, these measures correlate poorly with the in vivo behavior of transfused platelets and cannot be used to select optimal donors. Since radiolabeled platelet recovery and survival studies are expensive and time consuming, there is an ongoing search for simpler measures that predict platelet transfusion outcomes. We performed a pilot study using semi-qualitative proteomics to assess changes in the platelet protein profile of donors with either acceptable or unacceptable in vivo radiolabeled autologous platelet recovery and survival measurements. Proteins changing during a 9-day storage period included cytoskeletal elements talin, vinculin and moesin as well as signal transduction proteins 14-3-3, RhoGDI and Rap1. Two of nine donations exhibited a decrease in these proteins and poor in vivo platelet recovery and survival whereas the remaining donors showed acceptable platelet recovery and survival and expected protein profiles. Analyses revealed a significant correlation between protein levels of Rap1 and RhoGDI during storage and platelet recovery and survival. This study provides for the first time preliminary data showing evidence of the utility of protein profiling to predict platelet transfusion quality. This article is part of a Special Issue entitled: Integrated omics.

  7. Comparative analysis of platelet 5-HT concentrations in Han and Li patients with post-traumatic stress disorder.

    PubMed

    Li, L; Li, M X; Pan, L H; Wang, G M; Guo, M; Fu, L Q; Guo, J C; Gao, Y S; Chen, F; Xie, M X

    2016-07-15

    We investigated the role of serotonin (5-HT) in the pathogenesis of post-traumatic stress disorder (PTSD) by determining the platelet 5-HT concentrations in Li and Han patients with PTSD in Hainan Province, China. Li and Han control groups of the same sample size have no statistical differences in gender and age distribution compared to those in the PTSD groups who were also examined. The platelet 5-HT concentrations were determined by high-performance liquid chromatography. In addition, the patients and controls were evaluated by the impact of event scale-revised (IES-R). IES-R showed that the total and sub-scale scores of three factors (avoidance, intrusion, and hyperarousal) of Li patients with PTSD were significantly higher than those of Han patients with PTSD. Scores of both PTSD groups were higher than those of their respective control groups. The platelet 5-HT concentration of the Li patients with PTSD (120.56 ± 118.05 ng/10(9) platelets) was lower than that of the Han patients with PTSD (271.43 ± 181.66 ng/10(9) platelets) and that of both Li and Han control groups (338.54 ± 156.46, 350.58 ± 169.19 ng/10(9) platelets, respectively). Differences existed in symptoms of PTSD in terms of avoidance, intrusion, and hyperarousal in the Li and Han patients with PTSD. The diminished 5-HT activity in patients with PTSD may be relevant to biochemical changes in the brain and body. The differences in these factors between ethnic groups could be due to their customs, social status, and culture.

  8. Parallel comparison of apheresis-collected platelet concentrates stored in four different additive solutions.

    PubMed

    Nogawa, M; Naito, Y; Chatani, M; Onodera, H; Shiba, M; Okazaki, H; Matsuzaki, K; Satake, M; Nakajima, K; Tadokoro, K

    2013-11-01

    Partially replacing plasma with additive solutions in platelet (PLT) concentrates (PCs) may help to reduce transfusion reactions. Constituents of PLT additive solutions (PASs) have been revealed to affect the quality of PCs. Previous studies involved pairwise comparison of identical PLTs with two different PASs or multicomparison using random PLTs with three or more PASs. In this study, we performed parallel comparison using PCs from identical donors with four PASs. In addition to traditional parameters, the release of bioactive substances and plasma proteins was assessed. Platelets collected four times by apheresis from three donors were suspended in Intersol, SSP+, Composol or M-sol with 35% autologous plasma. The PC parameters, including PLT activation markers, glucose consumption, chemokines and plasma proteins, were assessed during 5-day storage. Mean PLT volumes were decreased in SSP+, Composol and M-sol after 5-day storage, with significant differences, whereas the hypertonic shock response (HSR) was decreased only in Intersol. Glucose consumption was faster in Intersol and M-sol than in SSP+ or Composol. PLT activation, determined as CD62P, sCD62P, sCD40L and RANTES, was significantly higher in Intersol than the other three PASs. No marked change was observed in fibrinopeptide A and C3a in any PASs. M-sol, SSP+ and Composol effectively preserved the quality of PCs. PLT activation was significantly enhanced in Intersol compared with the other three PASs. These effects seem to depend on magnesium and potassium as a constituent. Parallel comparison further verified that the PC quality largely depended on PASs but not donors. © 2013 International Society of Blood Transfusion.

  9. 21 CFR 864.9575 - Environmental chamber for storage of platelet concentrate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... to hold platelet-rich plasma within a preselected temperature range. (b) Classification. Class II... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Environmental chamber for storage of platelet... Establishments That Manufacture Blood and Blood Products § 864.9575 Environmental chamber for storage of...

  10. 21 CFR 864.9575 - Environmental chamber for storage of platelet concentrate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... to hold platelet-rich plasma within a preselected temperature range. (b) Classification. Class II... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Environmental chamber for storage of platelet... Establishments That Manufacture Blood and Blood Products § 864.9575 Environmental chamber for storage of...

  11. 21 CFR 864.9575 - Environmental chamber for storage of platelet concentrate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... to hold platelet-rich plasma within a preselected temperature range. (b) Classification. Class II... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Environmental chamber for storage of platelet... Establishments That Manufacture Blood and Blood Products § 864.9575 Environmental chamber for storage of...

  12. 21 CFR 864.9575 - Environmental chamber for storage of platelet concentrate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... to hold platelet-rich plasma within a preselected temperature range. (b) Classification. Class II... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Environmental chamber for storage of platelet... Establishments That Manufacture Blood and Blood Products § 864.9575 Environmental chamber for storage of...

  13. Supply with Platelet Concentrates from the Point of View of a Blood Donation Service of the Bavarian Red Cross.

    PubMed

    Weinauer, Franz

    2008-01-01

    SUMMARY: BACKGROUND: It is known that pooled platelet concentrates derived from buffy coat (PCs) have several disadvantages compared to platelet concentrates produced by platelet apheresis (APCs). Therefore, all blood products issued by the Bavarian Red Cross blood banks (BSD/BRK) (18,000 products/year) were produced by single donor apheresis. The main reason not to produce PCs was the elevated viral and bacterial infection risk during the last decade. But also the four-fold increased exposition to HLA and PLA antigens and the poor quality (in the sense of white and red cell contamination) of PCs (especially the ones produced with the platelet-rich plasma method) played a role to abstain from these products. MATERIAL AND METHODS: We performed a risk assessment to evaluate both products with regard to the actual testing and production methods, considering recently published data. However, a statistical calculation of the risks associated with the use of PCs or APCs with regard to different infectious agents with various prevalences was not done. RESULTS: The dramatically reduced risk for the transmission of HIV, HBV or HCV accompanying the implementation of improved antibody tests and of NAT minipool testing, the introduction of 100% leukocyte filtration, the conversion of PC production from the platelet-rich to the buffy coat method, and recent data on the risk of transmission of bacterial infections resulted in a equal assessment of APCs and PCs. CONCLUSION: As a consequence of this revised risk assessment, we supply our hospitals with both products APCs and buffy coat-derived PCs (pools of 4 donors). For clinical use we considered both products as equally effective, except for patients who have multiple antibodies and need HLA-typed platelets.

  14. Comparison between the clinical efficacy of platelet concentrates, derived from buffy coat and apheresis in tumor patients.

    PubMed

    Hao, Baolan; Wang, Yan; Zhou, Jian; Shao, Shujun; Dong, Xiaofeng

    2017-08-01

    The aim of the present study was to evaluate the clinical efficacy between manual buffy coat-derived platelet concentrates (PCs) and automated apheresis platelet concentrates (APCs) in terms of their therapeutic effects. The corrected count increment (CCI) was calculated according to detected differences in platelet concentration in patients who underwent transfusion of APCs, prepared by an automated system (group I, 72 cases) or PCs derived from buffy coat by manual method (group II, 83 cases). The clinical efficacy was assessed in terms of the CCI and clinical symptoms. The platelet contents of all the PCs were detected before transfusion. The mean 1 h CCI was 13.56±4.45 and 24 h CCI was 8.67±4.21 in group I, while the mean 1 h CCI was 15.83±4.65 and 24 h CCI was 9.57±3.36 in group II. The effective rates judged by CCI for groups I and II were 53 and 64%, respectively, and those judged by clinical symptoms were 67 and 60%, respectively. In conclusion, the clinical effectiveness of manual PCs was similar to that for APCs; thus, it could be utilized for clinical use.

  15. In vitro platelet function of platelet concentrates prepared using three different apheresis devices determined by impedance and optical aggregometry.

    PubMed

    Jilma-Stohlawetz, Petra; Eichelberger, Beate; Horvath, Michaela; Jilma, Bernd; Panzer, Simon

    2009-08-01

    Septic transfusion reactions to apheresis platelets (PLTs) continue to occur despite preventive measures. This study evaluated the effect of two operational changes designed to reduce bacterial risk: 1) introducing inlet-line sample diversion on two-arm procedures and 2) increasing the sample volume cultured from 4 to 8 mL from all donations. Aerobic culture results and septic transfusion reactions reported between December 1, 2006, and July 31, 2008 (Period 2), were compared to March 1, 2004, to May 31, 2006 (Period 1). During Period 2, a total of 781,936 apheresis PLT collections were cultured, of which 130 donations (1:6015) were confirmed positive and 9 (1:86,882) had negative culture results but were associated with 11 septic reactions. Confirmed-positive cultures from two-arm procedures decreased (27.2 to 14.7 per 10⁵ collections; odds ratio [OR], 0.54; 95% confidence interval [CI], 0.41-0.70) in Period 2, owing to a lower rate of skin flora contamination. Detection of contamination of one-arm collections significantly increased by 54% in Period 2 (13.7 vs. 21.1 per 10⁵ collections; OR, 1.54; 95% CI, 1.05-2.27). Fewer septic transfusion reactions occurred in Period 2, but the difference did not reach significance (1.7 vs. 1.2 per 10⁵ donations; OR, 0.68; 95% CI, 0.30-1.53). Inlet-line diversion decreased bacterial contamination during two-arm collections by more than 46%. Concurrently, doubling the sample volume was associated with a 54% relative increase in culture sensitivity. These interventions act cooperatively to decrease bacterial risk.

  16. Treatment of platelet concentrates with ultraviolet C light for pathogen reduction increases cytokine accumulation.

    PubMed

    Sandgren, Per; Berlin, Gösta; Tynngård, Nahreen

    2016-06-01

    Pathogen reduction technologies use photoactive substances in combination with ultraviolet (UV) light to inactivate pathogens. A new method uses only UVC light for pathogen reduction. This study assesses the effects of UVC light treatment on cytokine release in platelet (PLT) concentrates (PCs). A PC with 35% plasma and 65% PLT additive solution (SSP+) was prepared from five buffy coats. Three such PCs were pooled and divided into 3 units. One unit was used as a nonirradiated control, the second was a gamma-irradiated control, and the third unit was treated with UVC light technology. Ten units of each type were investigated. Cytokine release was analyzed on Days 1, 5, and 7 of storage. Correlation between cytokines, PLT surface markers, and hemostatic properties was investigated. Swirling was well preserved and pH was above the reference limit of 6.4 during storage of PLTs in all groups. Cytokine levels increased during storage in all groups but to a larger degree in PCs treated with UVC light. Only weak correlation was found between cytokines and PLT surface markers (r < 0.5). However, several cytokines showed strong correlation (r > 0.6) with the PLTs' ability to promote clot retraction. UVC treatment resulted in increased release from PLT alpha granules as evident by a higher cytokine release compared to nonirradiated and gamma-irradiated PCs. The clinical relevance of these findings needs to be further evaluated. © 2016 AABB.

  17. Prevalence of bacterial contamination in platelet concentrates at the National Center of Blood Transfusion (Mexico).

    PubMed

    Ibáñez-Cervantes, G; Bello-López, J M; Fernández-Sánchez, V; Domínguez-Mendoza, C A; Acevedo-Alfaro, L I

    2017-06-01

    Most common bacterial sepsis associated with transfusion is caused by contaminated Platelet Concentrates (PC). The screening of PC to detect bacterial contamination is obligatory in Mexico, and it is carried out in quality control programs. In Mexico, the identification and molecular characterization of bacterial contaminants to detect contamination sources have not been implemented due to high costs; however, it is an actual current need. One hundred PC were randomly selected and microbiologically analyzed. This sample size corresponds to 1% of the PC obtained by the National Center of Blood Transfusion (NCBT) in Mexico City according to the Official Mexican Standard NOM-253-SSA1-2012. Additionally, molecular biology tests were implemented in order to identify the possible contamination sources. Nine of the 100 PC analyzed (9%) showed bacterial contamination; analysis of the nucleotide sequences revealed the presence of characteristic microbiota from donor skin and soil. Diverse clonal relationship between the strains was identified in Staphylococcus epidermidis. Detection of contaminants associated with environmental and skin flora, shows the need to implement measures in the process of disinfecting skin at the site of phlebotomy and cleaning each of the areas involved in blood collection. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  18. INVESTIGATION OF BIOFILM FORMATION IN COAGULASE-NEGATIVE STAPHYLOCOCCI ISOLATED FROM PLATELET CONCENTRATE BAGS.

    PubMed

    Martini, Rosiéli; Hörner, Rosmari; Rampelotto, Roberta Filipini; Garzon, Litiérri Razia Litiérri; Nunes, Melise Silveira; Teixeira, Mayza Dalcin; Graichen, Daniel Ângelo Sganzerla

    2016-01-01

    Platelet Concentrates (PCs) are the blood components with the highest rate of bacterial contamination, and coagulase-negative staphylococci (CoNS) are the most frequently isolated contaminants. This study investigated the biofilm formation of 16 contaminated units out of 691 PCs tested by phenotypic and genotypic methods. Adhesion in Borosilicate Tube (ABT) and Congo Red Agar (CRA) tests were used to assess the presence of biofilm. The presence of icaADC genes was assessed by means of the Polymerase Chain Reaction (PCR) technique. With Vitek(r)2, Staphylococcus haemolyticus was considered the most prevalent CoNS (31.25%). The CRA characterized 43.8% as probable biofilm producers, and for the ABT test, 37.5%. The icaADC genes were identified in seven samples by the PCR. The ABT technique showed 85.7% sensitivity and 100% specificity when compared to the reference method (PCR), and presented strong agreement (k = 0.8). This study shows that species identified as PCs contaminants are considered inhabitants of the normal skin flora and they might become important pathogens. The results also lead to the recommendation of ABT use in laboratory routine for detecting biofilm in CoNS contaminants of PCs.

  19. INVESTIGATION OF BIOFILM FORMATION IN COAGULASE-NEGATIVE STAPHYLOCOCCI ISOLATED FROM PLATELET CONCENTRATE BAGS

    PubMed Central

    MARTINI, Rosiéli; HÖRNER, Rosmari; RAMPELOTTO, Roberta Filipini; GARZON, Litiérri Razia Litiérri; NUNES, Melise Silveira; TEIXEIRA, Mayza Dalcin; GRAICHEN, Daniel Ângelo Sganzerla

    2016-01-01

    Platelet Concentrates (PCs) are the blood components with the highest rate of bacterial contamination, and coagulase-negative staphylococci (CoNS) are the most frequently isolated contaminants. This study investigated the biofilm formation of 16 contaminated units out of 691 PCs tested by phenotypic and genotypic methods. Adhesion in Borosilicate Tube (ABT) and Congo Red Agar (CRA) tests were used to assess the presence of biofilm. The presence of icaADC genes was assessed by means of the Polymerase Chain Reaction (PCR) technique. With Vitek(r)2, Staphylococcus haemolyticus was considered the most prevalent CoNS (31.25%). The CRA characterized 43.8% as probable biofilm producers, and for the ABT test, 37.5%. The icaADC genes were identified in seven samples by the PCR. The ABT technique showed 85.7% sensitivity and 100% specificity when compared to the reference method (PCR), and presented strong agreement (k = 0.8). This study shows that species identified as PCs contaminants are considered inhabitants of the normal skin flora and they might become important pathogens. The results also lead to the recommendation of ABT use in laboratory routine for detecting biofilm in CoNS contaminants of PCs. PMID:26910444

  20. Optimization of the management of platelet concentrate stocks in the Basque Country using mathematical simulation.

    PubMed

    Pérez Vaquero, M Á; Gorria, C; Lezaun, M; López, F J; Monge, J; Eguizabal, C; Vesga, M A

    2016-05-01

    The management of platelet concentrate (PC) stocks is not simple given their short shelf life and variable demand. In general, managers decide on PC production based on personal experience. The objective of this study was to provide a tool to help decide how many PC units to produce each day in a more rational and objective way. From the historical data on PCs produced, transfused and discarded in the Basque Country in 2012, a mathematical model was built, based on the normality of the time series of the transfusions performed on each day of the week throughout the year. This model was implemented in an easy-to-use Excel spreadsheet and validated using real production data from 2013. Comparing with real 2013 data, in the best scenario, the number of PC units that expired was 87·7% lower, PC production, 14·3% lower and the age of the PCs transfused nearly 1-day younger in the simulation. If we want to ensure a minimum stock at the end of each day, the outdating rate and average age of the transfused PCs progressively increase. The practical application of the designed tool can facilitate decision-making about how many PC units to produce each day, resulting in very significant reductions in PC production and wastage and corresponding cost savings, together with an almost 1 day decrease in the mean age of PCs transfused. © 2016 The Authors. Vox Sanguinis published by John Wiley & Sons Ltd on behalf of International Society of Blood Transfusion.

  1. Bioresponsive polymers for the detection of bacterial contaminations in platelet concentrates.

    PubMed

    Gamerith, Clemens; Heinzle, Andrea; Schneider, Konstantin P; Hulla-Gumbsch, Elisabeth; Gewessler, Ulrike; Ducoroy, Laurent; Gehrer, Michael; Wagner, Thomas; Sigl, Eva; Guebitz, Georg M

    2014-03-25

    Bacterial contamination of platelet concentrates (PCs) can lead to fatal transfusion transmitted diseases and is the most abundant infectious risk in transfusion medicine. The storage conditions of PCs provide a good environment for bacterial growth. The detection of these contaminations at an early stage is therefore important to avoid the transfusion of contaminated samples. In this study, bioresponsive polymer (BRP) systems were used for the detection of microorganisms in PCs. The backbone of the polymer consisted of labelled protein (casein), which was demonstrated to be degraded by pure proteases as models and by extracellular enzymes released by contaminating microorganisms. The concomitant colour change was easily visible to the naked eye. To enhance stability, the protein was cross-linked with glycidyl methacrylate (GMA). The cross-linked polymer was easier to handle but was less sensitive than the non-cross-linked material. A contamination of a PC with 10CFU/mL S. aureus was detectable after 24 hours. The visible colour reaction was quantified as a ΔE value according to the CIELab concept. A ΔE value of 21.8 was already reached after 24 hours. Hence, this simple but effective system could prevent transfusion of a contaminated PC. Copyright © 2013. Published by Elsevier B.V.

  2. Platelet concentrates from fresh or overnight-stored blood, an international study.

    PubMed

    Dijkstra-Tiekstra, M J; van der Meer, P F; Cardigan, R; Devine, D; Prowse, C; Sandgren, P; de Wildt-Eggen, J

    2011-01-01

    Whole blood and also buffy coats (BCs) can be held for a few hours or overnight before processing into blood components or platelet concentrates (PCs). Individual studies have reported a range of outcomes regarding in vitro variables for PCs prepared from fresh and stored whole blood. In this multicenter study, effects of storage of whole blood or BCs on the in vitro quality of PCs were studied. The leukoreduced BC PCs were prepared from fresh BCs (2-8 hr after collection; fresh/fresh), from BCs at 20 to 24 hours after collection (fresh/stored), or from BCs prepared from whole blood stored for 20 to 24 hours (stored/fresh). PCs were stored on a flat-bed shaker at 20 to 24°C for 7 days. PCs were tested on Days 0 (only fresh/fresh), 1, 5, and 7 for in vitro quality. There were six participating centers that tested all three conditions with n = 6 per condition. In comparison to fresh/stored and stored/fresh PCs, fresh/fresh PCs exhibited a lower platelet (PLT) count (Day 1-220 × 10(9) ± 70 × 10(9) vs. 324 × 10(9) ± 50 × 10(9) and 368 × 10(9) ± 56 × 10(9) PLTs/PC), lactate, pCO(2), and hypotonic shock response (HSR; Days 5 and 7; Day 7-50 ± 13% vs. 57 ± 12 and 63 ± 11%) and a higher pH, glucose, pO(2), and CD62P expression (than stored/fresh PCs only; Day 7-33 ± 10% vs. 28 ± 12 and 24 ± 11%; p < 0.05). No differences were observed for volume, swirling effect, white blood cell count, annexin V binding, or aggregation between these conditions. Based on PLT count, HSR, and PLT activation, PCs are best prepared after 20 to 24 hours hold of the whole blood or BCs. © 2010 American Association of Blood Banks.

  3. Soluble Mediators in Platelet Concentrates Modulate Dendritic Cell Inflammatory Responses in an Experimental Model of Transfusion.

    PubMed

    Perros, Alexis J; Christensen, Anne-Marie; Flower, Robert L; Dean, Melinda M

    2015-10-01

    The transfusion of platelet concentrates (PCs) is widely used to treat thrombocytopenia and severe trauma. Ex vivo storage of PCs is associated with a storage lesion characterized by partial platelet activation and the release of soluble mediators, such as soluble CD40 ligand (sCD40L), RANTES, and interleukin (IL)-8. An in vitro whole blood culture transfusion model was employed to assess whether mediators present in PC supernatants (PC-SNs) modulated dendritic cell (DC)-specific inflammatory responses (intracellular staining) and the overall inflammatory response (cytometric bead array). Lipopolysaccharide (LPS) was included in parallel cultures to model the impact of PC-SNs on cell responses following toll-like receptor-mediated pathogen recognition. The impact of both the PC dose (10%, 25%) and ex vivo storage period was investigated [day 2 (D2), day 5 (D5), day 7 (D7)]. PC-SNs alone had minimal impact on DC-specific inflammatory responses and the overall inflammatory response. However, in the presence of LPS, exposure to PC-SNs resulted in a significant dose-associated suppression of the production of DC IL-12, IL-6, IL-1α, tumor necrosis factor-α (TNF-α), and macrophage inflammatory protein (MIP)-1β and storage-associated suppression of the production of DC IL-10, TNF-α, and IL-8. For the overall inflammatory response, IL-6, TNF-α, MIP-1α, MIP-1β, and inflammatory protein (IP)-10 were significantly suppressed and IL-8, IL-10, and IL-1β significantly increased following exposure to PC-SNs in the presence of LPS. These data suggest that soluble mediators present in PCs significantly suppress DC function and modulate the overall inflammatory response, particularly in the presence of an infectious stimulus. Given the central role of DCs in the initiation and regulation of the immune response, these results suggest that modulation of the DC inflammatory profile is a probable mechanism contributing to transfusion-related complications.

  4. In vitro study of platelet function confirms the contribution of the ultraviolet B (UVB) radiation in the lesions observed in riboflavin/UVB-treated platelet concentrates.

    PubMed

    Abonnenc, Mélanie; Sonego, Giona; Crettaz, David; Aliotta, Alessandro; Prudent, Michel; Tissot, Jean-Daniel; Lion, Niels

    2015-09-01

    Platelet inactivation technologies (PITs) have been shown to increase platelet storage lesions (PSLs). This study investigates amotosalen/ultraviolet (UV)A- and riboflavin/UVB-induced platelet (PLT) lesions in vitro. Particular attention is given to the effect of UVB alone on PLTs. Buffy coat-derived PLT concentrates (PCs) were treated with amotosalen/UVA, riboflavin/UVB, or UVB alone and compared to untreated PCs throughout storage. In vitro PLT function was assessed by blood gas and metabolite analyses, flow cytometry-based assays (CD62P, JC-1, annexin V, PAC-1), hypotonic shock response, and static adhesion to fibrinogen-coated wells. In our experimental conditions, riboflavin/UVB-treated PCs showed the most pronounced differences compared to untreated and amotosalen/UVA-treated PCs. The riboflavin/UVB treatment led to a significant increase of anaerobic glycolysis rate despite functional mitochondria, a significant increase of CD62P on Day 2, and a decrease of JC-1 aggregates and increase of annexin V on Day 7. The expression of active GPIIbIIIa (PAC-1) and the adhesion to fibrinogen was significantly increased from Day 2 of storage in riboflavin/UVB-treated PCs. Importantly, we showed that these lesions were caused by the UVB radiation alone, independently of the presence of riboflavin. The amotosalen/UVA-treated PCs confirmed previously published results with a slight increase of PSLs compared to untreated PCs. Riboflavin/UVB-treated PCs present significant in vitro PSLs compared to untreated PCs. These lesions are caused by the UVB radiation alone and probably involve the generation of reactive oxygen species. The impact of these observations on clinical use must be investigated. © 2015 AABB.

  5. Mechanical and degradation properties of advanced platelet-rich fibrin (A-PRF), concentrated growth factors (CGF), and platelet-poor plasma-derived fibrin (PPTF).

    PubMed

    Isobe, Kazushige; Watanebe, Taisuke; Kawabata, Hideo; Kitamura, Yutaka; Okudera, Toshimitsu; Okudera, Hajime; Uematsu, Kohya; Okuda, Kazuhiro; Nakata, Koh; Tanaka, Takaaki; Kawase, Tomoyuki

    2017-12-01

    Fibrin clot membranes prepared from advanced platelet-rich fibrin (A-PRF) or concentrated growth factors (CGF), despite their relatively rapid biodegradability, have been used as bioactive barrier membranes for alveolar bone tissue regeneration. As the membranes degrade, it is thought that the growth factors are gradually released. However, the mechanical and degradable properties of these membranes have not well been characterized. The purpose of this study was to mechanically and chemically characterize these membranes. A-PRF and CGF clots were prepared from blood samples collected from non-smoking, healthy donors and were compressed to form 1-mm-thick membranes. Platelet-poor plasma-derived fibrin (PPTF) clots were prepared by adding bovine thrombin to platelet-poor plasma. A tensile test was performed at the speed of 1 mm/min. Morphology of the fibrin fibers was examined by SEM. A digestion test was performed in PBS containing trypsin and EDTA. In the tensile test, statistical difference was not observed in Young's modulus, strain at break, or maximum stress between A-PRF and CGF. In strain at break, PPTF was significantly weaker than CGF. Likewise, fibrin fiber thickness and crosslink density of PPTF were less than those of other membranes, and PPTF degraded faster than others. Although the centrifugal conditions are different, A-PRF and CGF are prepared by essentially identical mechanisms. Therefore, it is conceivable that both membranes have similar mechanical and chemical properties. Only PPTF, which was prepared by a different mechanism, was characterized as mechanically weaker and enzymatically more degradable.

  6. Plasma and serum serotonin concentrations and surface-bound platelet serotonin expression in Cavalier King Charles Spaniels with myxomatous mitral valve disease.

    PubMed

    Cremer, Signe E; Kristensen, Annemarie T; Reimann, Maria J; Eriksen, Nynne B; Petersen, Stine F; Marschner, Clara B; Tarnow, Inge; Oyama, Mark A; Olsen, Lisbeth H

    2015-06-01

    To investigate serum and plasma serotonin concentrations, percentage of serotonin-positive platelets, level of surface-bound platelet serotonin expression (mean fluorescence intensity [MFI]), and platelet activation (CD62 expression) in platelet-rich plasma from Cavalier King Charles Spaniels with myxomatous mitral valve disease (MMVD). Healthy dogs (n = 15) and dogs with mild MMVD (18), moderate-severe MMVD (19), or severe MMVD with congestive heart failure (CHF; 10). Blood samples were collected from each dog. Serum and plasma serotonin concentrations were measured with an ELISA, and surface-bound platelet serotonin expression and platelet activation were determined by flow cytometry. Dogs with mild MMVD had higher median serum (746 ng/mL) and plasma (33.3 ng/mL) serotonin concentrations, compared with MMVD-affected dogs with CHF (388 ng/mL and 9.9 ng/mL, respectively), but no other group differences were found. Among disease groups, no differences in surface-bound serotonin expression or platelet activation were found. Thrombocytopenic dogs had lower serum serotonin concentration (482 ng/mL) than nonthrombocytopenic dogs (731 ng/mL). In 26 dogs, a flow cytometry scatterplot subpopulation (FSSP) of platelets was identified; dogs with an FSSP had a higher percentage of serotonin-positive platelets (11.0%), higher level of surface-bound serotonin expression (MFI, 32,068), and higher platelet activation (MFI, 2,363) than did dogs without an FSSP (5.7%, 1,230, and 1,165, respectively). An FSSP was present in 93.8% of thrombocytopenic dogs and in 29.5% of nonthrombocytopenic dogs. A substantive influence of circulating serotonin on MMVD stages prior to CHF development in Cavalier King Charles Spaniels was not supported by the study findings. An FSSP of highly activated platelets with pronounced serotonin binding was strongly associated with thrombocytopenia but not MMVD.

  7. Monitoring the intracellular store Ca2+ concentration in agonist-stimulated, intact human platelets by using Fluo-5N.

    PubMed

    Sage, S O; Pugh, N; Mason, M J; Harper, A G S

    2011-03-01

    Most Ca(2+) signaling research in platelets has relied solely on monitoring the cytosolic Ca(2+) concentration ([Ca(2+)](cyt)). Changes in [Ca(2+)](cyt) constitute the net effect of Ca(2+) fluxes into the cytosol across the plasma membrane (PM) and from intracellular stores, and Ca(2+) sequestration into the stores and Ca(2+) removal across the PM. This makes interpretation of the effects of pharmacologic or genetic interventions on Ca(2+) signaling difficult and subject to error. To validate the use of the low-affinity Ca(2+) indicator Fluo-5N to monitor the concentration of Ca(2+) in the intracellular stores ([Ca(2+)](st)) of human platelets as a first step in developing assays for a systems-level analysis of platelet Ca(2+) signaling. Fluo-5N-loaded and Fura-2-loaded human platelets were used to observe the effects of agonist stimulation and other manipulations on [Ca(2+)](cyt) and [Ca(2+)](st). Fluo-5N fluorescence changed appropriately in response to compounds that induce passive depletion of intracellular Ca(2+) stores and to physiologic agonists. Ca(2+) reuptake inhibitors and blockers of Ca(2+) release channels had the expected effects on Fura-2 and Fluo-5N fluorescence. Agonist-evoked Ca(2+) release was reversed by Ca(2+) addition to the medium, and required intact Ca(2+) reuptake mechanisms. Store refilling was observed in the presence of sarcoplasmic/endoplasmic reticulum Ca(2+) -ATPase (SERCA) inhibitors and ionomycin, suggesting the presence of a non-SERCA Ca(2+) reuptake mechanism. Evidence for a role for Ca(2+) -induced Ca(2+) release in agonist-evoked responses was obtained. Our data provide a validation of the use of Fluo-5N as a method for monitoring changes in [Ca(2+)](st) in human platelets. © 2011 International Society on Thrombosis and Haemostasis.

  8. Feasibility and usefulness of self-assessment of bleeding in patients with haematological malignancies, and the association between platelet count and bleeding.

    PubMed

    Stanworth, S J; Dyer, C; Casbard, A; Murphy, M F

    2006-07-01

    The aim of this study was to evaluate the collection of daily prospective information about bleeding outcomes in patients with thrombocytopenia, including information obtained by patient self-assessment. Consecutive patients with haematological malignancies were enrolled in a study of bleeding data collection during the period of thrombocytopenia. A short educational session and information sheet was designed for self-assessment. Platelet counts and all transfusions were recorded daily. Bleeding scores were translated into World Health Organization (WHO) bleeding grades. Nineteen patients were included in the study. Four-hundred and ten days of thrombocytopenia were eligible for assessment of bleeds. Self-assessment was feasible, as defined by the total proportion of days on which self-assessment was completed (70%, 288 thrombocytopenic days). There was 86% agreement between bleeding data collected by self-assessment and by medical examination using a structured assessment form. Examples of discrepancies included the duration of petechiae/bruises and the reporting of minor bleeding. There was no evidence for an association between patients' morning platelet count and daily WHO bleeding grade. The incidences of WHO grade 1 and grade 2 bleeding on days with platelet counts < or = 10 x 10(9)/l, 11-20 x 10(9)/l, and > 20 x 10(9)/l were similar and did not reveal higher rates of bleeding at lower counts. Patient self-assessment can help to support comprehensive daily prospective monitoring of bleeding, specifically facilitating data collection following hospital discharge. The discrepancies between self-assessment and medical examination highlight the need to develop a validated international assessment tool. The association among platelet count, risk of bleeding and role of prophylactic platelet transfusions needs further evaluation in larger prospective trials.

  9. Comparison of platelet-rich plasma (PRP), platelet-rich fibrin (PRF), and concentrated growth factor (CGF) in rabbit-skull defect healing.

    PubMed

    Kim, Tae-Hoon; Kim, Sung-Hee; Sándor, George K; Kim, Yong-Deok

    2014-05-01

    The objective of this study was to evaluate the effect of platelet-rich plasma (PRP), platelet-rich fibrin (PRF), and concentrated growth factor (CGF) on bone healing. Twelve rabbits were included in this randomized, blinded, prospective study. 15-mm×10-mm-sized defects were created in the parietal bone, filled with PRP, PRF, CGF, and void. The bone mineral density and bone volume were analyzed with microscopic computed tomography (micro-CT) and histomorphometrics at the 6th and 12th week. In micro-CT analysis, bone mineral density and bone volume were greater in the experimental group than in controls at both 6th and 12th week, but not among the experimental groups. Similarly, histomorphometric examination revealed that more bone formation was seen in the experimental group. The addition of PRP, PRF, and CGF had significantly increased bone formation at the 6th week. The effect of PRP, PRF, and CGF was similar and may be useful in the future to increase the success rate of bone grafting. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. [Transfusion-transmitted bacterial infection of a apheresis platelet concentrate with Streptococcus gallolyticus: Analysis of one case].

    PubMed

    Le Niger, C; Dalbies, F; Narbonne, V; Hery-Arnaud, G; Virmaux, M; Léostic, C; Hervé, F; Liétard, C

    2014-06-01

    Bacterial infections are uncommon complications of the blood products transfusion but they are potentially serious. Many advances have been done over the past few years to guarantee the microbiological security of blood products as the donors selection with a medical talk, the derivation of the first 30 millilitres blood during the donation, the deleucocytation of blood products… But in spite of these advances, cases of bacterial infection always remain. The purpose of this study was to point out the platelet concentrate's transfusion-transmitted bacterial infection with Streptococcus gallolyticus and the unusual consequence for the donor by uncovering an asymptomatic rectal neoplastic tumor. This study as raised as to whether the usefulness of systematic bacterial inactivation in the platelets concentrates.

  11. The effects of aspirin on platelet function and lysophosphatidic acids depend on plasma concentrations of EPA and DHA.

    PubMed

    Block, Robert C; Abdolahi, Amir; Tu, Xin; Georas, Steve N; Brenna, J Thomas; Phipps, Richard P; Lawrence, Peter; Mousa, Shaker A

    2015-05-01

    Aspirin's prevention of cardiovascular disease (CVD) events in individuals with type 2 diabetes mellitus is controversial. Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) and aspirin all affect the cyclooxygenase enzyme. The relationship between plasma EPA and DHA and aspirin's effects has not been determined. Thirty adults with type 2 diabetes mellitus ingested aspirin (81 mg/day) for 7 days, then EPA+DHA (2.6g/day) for 28 days, then both for another 7 days. Lysophosphatidic acid (LPA) species and more classic platelet function outcomes were determined. Plasma concentrations of total EPA+DHA were associated with 7-day aspirin reduction effects on these outcomes in a "V"-shaped manner for all 11 LPA species and ADP-induced platelet aggregation. This EPA+DHA concentration was quite consistent for each of the LPA species and ADP. These results support aspirin effects on lysolipid metabolism and platelet aggregation depending on plasma EPA+DHA concentrations in individuals with a disturbed lipid milieu. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. The effects of aspirin on platelet function and lysophosphatidic acids depend on plasma concentrations of EPA and DHA

    PubMed Central

    Block, Robert C; Abdolahi, Amir; Tu, Xin; Georas, Steve N; Brenna, J. Thomas; Phipps, Richard P; Lawrence, Peter; Mousa, Shaker A

    2015-01-01

    Aspirin’s prevention of cardiovascular disease (CVD) events in individuals with type 2 diabetes mellitus is controversial. Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) and aspirin all affect the cyclooxygenase enzyme. The relationship between plasma EPA and DHA and aspirin’s effects has not been determined. Thirty adults with type 2 diabetes mellitus ingested aspirin (81 mg/day) for 7 days, then EPA+DHA (2.6 g/day) for 28 days, then both for another 7 days. Lysophosphatidic acid (LPA) species and more classic platelet function outcomes were determined. Plasma concentrations of total EPA+DHA were associated with 7-day aspirin reduction effects on these outcomes in a “V”-shaped manner for all 11 LPA species and ADP-induced platelet aggregation. This EPA+DHA concentration was quite consistent for each of the LPA species and ADP. These results support aspirin effects on lysolipid metabolism and platelet aggregation depending on plasma EPA+DHA concentrations in individuals with a disturbed lipid milieu. PMID:25555354

  13. A prospective, longitudinal study of platelet serotonin and plasma brain-derived neurotrophic factor concentrations in major depression: effects of vortioxetine treatment.

    PubMed

    Sagud, Marina; Nikolac Perkovic, Matea; Vuksan-Cusa, Bjanka; Maravic, Anja; Svob Strac, Dubravka; Mihaljevic Peles, Alma; Zivkovic, Maja; Kusevic, Zorana; Pivac, Nela

    2016-09-01

    Various antidepressants occupy brain serotonin transporter (SERT), decrease platelet serotonin (5-HT) concentration, and normalize reduced plasma brain-derived neurotrophic factor (BDNF) concentrations in depressed patients. Vortioxetine is a recently introduced antidepressant with a multimodal mechanism of action. In addition to SERT inhibition, vortioxetine acts via different 5-HT receptors. To further elucidate its mechanism of action, we have investigated the effects of vortioxetine on platelet 5-HT and plasma BDNF concentrations in patients with major depression. Platelet 5-HT and plasma BDNF concentrations were determined in 44 healthy subjects at baseline and in 44 depressed patients before and after 4 weeks of treatment with vortioxetine (5-15 mg daily). Platelet 5-HT concentration was determined using the ortho-phthalaldehyde-enhanced fluorometric method, and plasma BDNF concentration using a commercial enzyme-linked immunosorbent assay (Quantikine ELISA, R&D Systems). At baseline, platelet 5-HT concentrations did not differ between depressed and control subjects, but plasma BDNF values were lower (p = 0.011; ω = 0.80) in depressed patients than in healthy subjects. Vortioxetine treatment significantly (p < 0.0001; ω = 0.80) decreased platelet 5-HT concentration and significantly (p = 0.004; ω = 0.80) increased plasma BDNF concentration in depressed patients compared to their baseline values. Age, gender, and smoking were not significantly associated with platelet 5-HT and plasma BDNF concentrations. Despite a novel mechanism of action, vortioxetine shares some common effects with other antidepressants. This study is the first to show that, in addition to clinical improvement, 4 weeks of treatment with vortioxetine (5-15 mg daily), decreased platelet 5-HT and increased plasma BDNF concentrations in depressed patients.

  14. Healing of Postextraction Sockets Preserved With Autologous Platelet Concentrates. A Systematic Review and Meta-Analysis.

    PubMed

    Del Fabbro, Massimo; Bucchi, Cristina; Lolato, Alessandra; Corbella, Stefano; Testori, Tiziano; Taschieri, Silvio

    2017-08-01

    The true benefit of autologous platelet concentrates (APCs) for enhancing the healing of postextraction sites is still a matter of debate, and in recent years several clinical trials have addressed this issue. The purpose of this study was to determine the effectiveness of an APC adjunct in the preservation of fresh extraction sockets. An electronic search was performed on Medline, Embase, Scopus, and the Cochrane Central Register of Controlled Trials. Only controlled clinical trials or randomized clinical trials were included. Selected articles underwent risk-of-bias assessment. The outcomes were complications and adverse events, discomfort and quality of life, bone healing and remodeling assessed by histologic and radiographic techniques, and soft tissue healing. Thirty-three comparative studies were included. Nine articles had a parallel design and 24 had a split-mouth design. Twenty studies were considered to have a low risk of bias and 13 were considered to have a high risk. Overall, 1,193 teeth were extracted from 911 patients. Meta-analysis showed that soft tissue healing, probing depth at 3 months, and bone density at 1, 3, and 6 months were statistically better for the APC group. Qualitative analysis suggested that APCs might be associated with a decrease in swelling and trismus. However, no relevant difference among groups was found for probing depth at 1 month, incidence of alveolar osteitis, acute inflammation or infection, percentage of new bone, and indirect measurement of bone metabolism. APCs should be used in postextraction sites to improve clinical and radiographic outcomes such as bone density and soft tissue healing and postoperative symptoms. The actual benefit of APCs on decreasing pain in extraction sockets is still not quantifiable. Copyright © 2017 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.

  15. pH value promotes growth of Staphylococcus epidermidis in platelet concentrates.

    PubMed

    Störmer, Melanie; Kleesiek, Knut; Dreier, Jens

    2008-05-01

    The platelet (PLT) storage lesion is characterized metabolically by a pH value associated with lactic acid generation. PLT storage conditions support the growth of Staphylococcus epidermidis, the most common organism implicated in bacterial contamination of PLT concentrates (PCs). Here, different factors that influence bacterial growth in PCs are discussed and the relation between pH values of PCs and citrate plasma (CP) is studied, with emphasis on bacterial proliferation. The PLT lesion with regard to pH decrease and lactic acid production was monitored during storage and correlated to bacterial proliferation properties. A total of 115 coagulase-negative staphylococci, especially S. epidermidis isolates, were characterized for their proliferation in different blood components (CP, buffy coat-derived, and apheresis PCs). Furthermore, the influence of donor-specific, product-specific, species-specific, and strain-specific factors on bacterial proliferation was investigated. PCs showed a lower pH value in comparison to plasma during storage. Bacterial proliferation in PCs and the failure to grow in CP were determined with all organisms tested. No correlation to donor-specific, species-specific, or strain-specific factors was observed. Lowering the pH of CP resulted in bacterial proliferation, whereas a pH increase in the PC unit inhibited the proliferation of S. epidermidis. With emphasis on bacterial proliferation, the significant difference between PC and CP is the presence of metabolizing PLTs. The pH values of stored PLTs, but not those of stored plasma, support the growth of S. epidermidis.

  16. Autologous platelet concentrate in surgery for macular detachment associated with congenital optic disc pit

    PubMed Central

    Nadal, Jeroni; Figueroa, Marta S; Carreras, Elisa; Pujol, Patricia; Canut, Maria Isabel; Barraquer, Rafael Ignacio

    2015-01-01

    Purpose To evaluate the anatomical and functional results obtained with pars plana vitrectomy (PPV) plus autologous platelet concentrate (APC) as a treatment for macular detachment associated with optic disc pit (ODP). Methods We performed a prospective interventional study of 19 eyes of 19 consecutive patients with posterior macular detachment due to ODP. All patients underwent PPV, posterior hyaloid peeling, fluid–air exchange, injection of 0.05 mL of APC over the ODP and 15% perfluoropropane (C3F8) endotamponade. Postoperative measures included face-up positioning for 2 hours and then avoidance of the face-up position during the ensuing 10 days. All patients underwent complete ophthalmologic examination and optical coherence tomography preoperatively at 1 month, 3 months, 6 months, 9 months, and 12 months postoperatively and then annually. Outcome measures were best corrected visual acuity (BCVA) by logMAR, improvement of quality of vision, macular attachment, and resolution of intraretinal schisis-like separation. Results Preoperatively, the median BCVA was 0.70 (range: 0.30–1.70) and all patients showed improved visual acuity after surgery; BCVA was 0.22 (range: 0.07–0.52) at 12 months follow-up. All patients showed complete reabsorption of intraretinal fluid (median time: 3.5 months [range: 2–8 months]) and macular attachment at the end of follow-up (median: 60 months [range: 12–144 months]), with stable or improved visual acuity. No reoperations were needed and no major adverse events were recorded. Conclusion For macular detachment associated with ODP, the combination of PPV, posterior hyaloid peeling, APC, and C3F8 tamponade is a highly effective alternative technique with stable anatomical and functional results. PMID:26543348

  17. A new one-platform flow cytometric method for residual cell counting in platelet concentrates.

    PubMed

    Schmidt, Michael; Spengler, Hans-Peter; Lambrecht, Bernd; Hourfar, Michael K; Seifried, Erhard; Tonn, Torsten

    2009-12-01

    According to German regulations and guidelines, residual red blood cells (rRBCs) and residual white blood cells (rWBCs) must number fewer than 3 x 10(9) cells/unit and 1 x 10(6) cells/unit in platelet concentrates (PCs), respectively. Due to low levels of residual cells in final products, there is still a need for fast, reliable, and sensitive methods of automated detection of these cell types. In Part A, 21 PCs were spiked with predetermined numbers of red blood cells (RBCs) and white blood cells (WBCs). The linearity, precision, and accuracy of the BD Thrombo Count assay (BD Biosciences Europe) were tested and validated according to international guidelines. Finally in Part B, 100 PCs prepared from pooled buffy coats were tested by the BD Thrombo Count assay and compared with other methods, including Nageotte (rWBCs) and Neubauer (rRBCs) counting chambers and the flow cytometric BD LeucoCOUNT (Becton Dickinson) assay (rWBCs). The unspecific background of blank PC samples was fewer than 0.02 cells/microL for WBCs and fewer than 34 cells/microL for RBCs (mean, 21). Linear regression and precision analyses of spiked PC samples were determined for both WBCs (r(2) = 0.992; range, 0.6-6.0 WBCs/microL) and RBCs (r(2) = 0.999; 800-8000 RBCs/microL). No carryover of cells or drift in results was detected in the automated sample acquisition mode. Analysis according to statistical methods of Bland and Altman demonstrated a high correlation between BD Thrombo Count and the Neubauer manual counting chamber. This novel flow cytometric test is a quick and reliable single-tube assay that has been demonstrated as a potential alternative for the existing manual microscopic counting procedures that are both time-consuming and laborious.

  18. Plasma concentrations of endotoxin and platelet activation in the developmental stage of oligofructose-induced laminitis.

    PubMed

    Bailey, S R; Adair, H S; Reinemeyer, C R; Morgan, S J; Brooks, A C; Longhofer, S L; Elliott, J

    2009-06-15

    The link between the fermentation of carbohydrate in the equine large intestine and the development of acute laminitis is poorly understood. Absorption of endotoxin (lipopolysaccharide; LPS) into the plasma has been observed in one experimental model of laminitis, but does not cause laminitis when administered alone. Thus, the potential role of endotoxin is unclear. Platelet activation has previously been demonstrated in the developmental stage of laminitis. Equine platelets are more sensitive than leukocytes to activation by endotoxin, and can be activated directly by LPS in the low pg/ml range, activating p38 MAP kinase and releasing serotonin (5-HT) and thromboxane. The objectives of this study were firstly to determine whether endotoxin and platelet activation could be measured in the plasma of horses in the developmental phase of laminitis induced with oligofructose. Secondly, the time course of events involving platelet activation and platelet-derived vasoactive mediator production was investigated. Laminitis was induced in six Standardbred horses by the administration of 10 g/kg bwt of oligofructose. Plasma samples were obtained every 4h, and platelet pellets were obtained by centrifugation. LPS was measured using a kinetic limulus amebocyte lysate assay, and platelet activation was assessed by Western blotting for the phosphorylated form of p38 MAP kinase. Plasma 5-HT was assayed by HPLC with electrochemical detection and thromboxane B(2) was measured by radioimmunoassay. Clinical signs of laminitis and histopathologic changes were observed in lamellar sections from five of the six horses. Onset of lameness was between 20 and 30 h after the administration of oligofructose. LPS increased above the limit of detection (0.6 pg/ml) to reach a peak of 2.4+/-1.0 pg/ml at 8 h. TNFalpha was also detectable in the plasma from 12 to 24 h. There was a time-dependent increase in platelet p38 MAPK phosphorylation, which peaked at approximately 12 h (3.8+/-1.3 fold

  19. Simultaneous concentration of platelets and marrow cells: a simple and useful technique to obtain source cells and growth factors for regenerative medicine.

    PubMed

    Nishimoto, Soh; Oyama, Tomoki; Matsuda, Ken

    2007-01-01

    Platelet-rich plasma (PRP) has attracted attention as a safe and cost-effective source of growth factors that stimulate cells to regenerate tissue. Bone marrow aspirate was processed with the same protocol to obtain PRP from peripheral blood. This concentrate contained condensed nucleated bone marrow cells, which are useful for regenerative medicine, as well as condensed platelets. In PRP derived from bone marrow aspirate, the density of platelets and levels of growth factors (platelet-derived growth factor and transforming growth factor-beta) were the same as in PRP derived from peripheral blood. Condensation of nucleated cells, especially small-sized cells, was confirmed. With a simple and cost-effective technique, source cells and growth factors can be obtained at the same time. This simultaneous concentration of platelets and bone marrow cells has great potential as a source of materials for regenerative medicine.

  20. In vitro evaluation of COM.TEC apheresis platelet concentrates using a preparation set and pathogen inactivation over a storage period of five days.

    PubMed

    Moog, R; Fröhlich, A; Mayaudon, V; Lin, L

    2004-01-01

    The aim of the present study was to evaluate in vitro data on platelets collected by apheresis, processed on a preparation set followed by photochemical treatment (PCT). Fifteen single-donor platelet concentrates (PCs) were collected by apheresis (COM.TEC blood cell separator, Fresenius, Bad Homburg, Germany). The platelets were transferred to the preparation set and plasma was removed after centrifugation to resuspend the platelets in approximately 37% plasma and 63% platelet additive solution InterSol. PCT was done by exposing the platelets to amotosalen HCl followed by illumination with ultraviolet light. Blood cell counts and in vitro PLT function were measured up to 5 days. An average of 3.44 +/- 0.28 x 10(11) platelets were collected in a product volume of 351 +/- 21 mL. Plasma removal resulted in a mean platelet loss of 7.8%. After PCT, a progressive decrease in platelet function was observed. LDH level rose through storage (171 +/- 81 U/L) to levels approximating LDH levels observed post-collection (180 +/- 103 U/L). There was a gradual decrease of the platelets to respond to hypotonic shock response from 90 +/- 9 % post-plasma reduction to 48 +/- 16% at day 5. All PLT units met the European requirements for leukoreduction and the pH limit of 6.8 up to day 5 post-collection. The new preparation set was capable of producing platelet units meeting the requirements for PCT. Despite differences observed in in vitro platelet function parameters, PLTs at storage day 5 fit the German and European guidelines.

  1. Blood platelet kinetics and platelet transfusion.

    PubMed

    Aster, Richard H

    2013-11-01

    The discovery of citrate anticoagulant in the 1920s and the development of plastic packs for blood collection in the 1960s laid the groundwork for platelet transfusion therapy on a scale not previously possible. A major limitation, however, was the finding that platelet concentrates prepared from blood anticoagulated with citrate were unsuitable for transfusion because of platelet clumping. We found that this could be prevented by simply reducing the pH of platelet-rich plasma to about 6.5 prior to centrifugation. We used this approach to characterize platelet kinetics and sites of platelet sequestration in normal and pathologic states and to define the influence of variables such as anticoagulant and ABO incompatibility on post-transfusion platelet recovery. The "acidification" approach enabled much wider use of platelet transfusion therapy until alternative means of producing concentrates suitable for transfusion became available.

  2. Blood platelet kinetics and platelet transfusion

    PubMed Central

    Aster, Richard H.

    2013-01-01

    The discovery of citrate anticoagulant in the 1920s and the development of plastic packs for blood collection in the 1960s laid the groundwork for platelet transfusion therapy on a scale not previously possible. A major limitation, however, was the finding that platelet concentrates prepared from blood anticoagulated with citrate were unsuitable for transfusion because of platelet clumping. We found that this could be prevented by simply reducing the pH of platelet-rich plasma to about 6.5 prior to centrifugation. We used this approach to characterize platelet kinetics and sites of platelet sequestration in normal and pathologic states and to define the influence of variables such as anticoagulant and ABO incompatibility on post-transfusion platelet recovery. The “acidification” approach enabled much wider use of platelet transfusion therapy until alternative means of producing concentrates suitable for transfusion became available. PMID:24177466

  3. Reduction of relative centrifugation force within injectable platelet-rich-fibrin (PRF) concentrates advances patients' own inflammatory cells, platelets and growth factors: the first introduction to the low speed centrifugation concept.

    PubMed

    Choukroun, J; Ghanaati, S

    2017-03-10

    The aim of this study was to analyze systematically the influence of the relative centrifugation force (RCF) on leukocytes, platelets and growth factor release within fluid platelet-rich fibrin matrices (PRF). Systematically using peripheral blood from six healthy volunteers, the RCF was reduced four times for each of the three experimental protocols (I-III) within the spectrum (710-44 g), while maintaining a constant centrifugation time. Flow cytometry was applied to determine the platelets and leukocyte number. The growth factor concentration was quantified 1 and 24 h after clotting using ELISA. Reducing RCF in accordance with protocol-II (177 g) led to a significantly higher platelets and leukocytes numbers compared to protocol-I (710 g). Protocol-III (44 g) showed a highly significant increase of leukocytes and platelets number in comparison to -I and -II. The growth factors' concentration of VEGF and TGF-β1 was significantly higher in protocol-II compared to -I, whereas protocol-III exhibited significantly higher growth factor concentration compared to protocols-I and -II. These findings were observed among 1 and 24 h after clotting, as well as the accumulated growth factor concentration over 24 h. Based on the results, it has been demonstrated that it is possible to enrich PRF-based fluid matrices with leukocytes, platelets and growth factors by means of a single alteration of the centrifugation settings within the clinical routine. We postulate that the so-called low speed centrifugation concept (LSCC) selectively enriches leukocytes, platelets and growth factors within fluid PRF-based matrices. Further studies are needed to evaluate the effect of cell and growth factor enrichment on wound healing and tissue regeneration while comparing blood concentrates gained by high and low RCF.

  4. Interaction of platelet-rich concentrate with bone graft materials: an in vitro study.

    PubMed

    Butcher, Andrew; Milner, Richard; Ellis, Keith; Watson, J Tracy; Horner, Alan

    2009-03-01

    Platelet-rich concentrate (PRC) is in routine use for orthopaedic and maxilofacial surgery and is frequently combined with bone graft materials to fill bony defects and enhance healing. Numerous studies have been performed investigating the efficacy of PRC to enhance bone healing in which a variety of graft materials have been combined with varying degrees of success. Here, we sought to determine the effect of combining PRC with different graft materials on human bone marrow stromal cell (hBMSC) proliferation, osteoblastic differentiation, and bone formation. Our central hypothesis is that PRC is not a true osteogenic agent but rather is osteopromotive, with cell fate determination being dependent on additional signals derived from the microenvironment. Experiments were performed with low passage (maximum 3) hBMSCs that were maintained in the presence of ascorbic acid-2-phosphate and beta-glycerol phosphate. Dexamethasone was excluded from these studies. PRC and graft materials were retained within well inserts and clotted by addition of bovine thrombin. Cell proliferation was determined by DNA content, osteoblastic commitment, and differentiation by alkaline phosphatase activity and matrix mineralization. Combining PRC with the graft materials increased proliferation above that seen with the graft materials alone; however, only demineralized bone matrix (DBM) and allograft were capable of increasing proliferation above that seen with PRC alone. The increased proliferation observed in the presence of PRC coincided with decreased normalized alkaline phosphatase activity, suggesting decreased osteoblastic differentiation. However, at later time points, PRC increased mineralization compared with DBM, collagen, or beta tricalcium phosphate alone. When compared with PRC alone, addition of DBM or allograft decreased mineralization. Collagen gave rise to a small increase in mineralization, whereas beta tricalcium phosphate yielded the same level of mineralization as PRC

  5. Quality assessment of buffy-coat-derived leucodepleted platelet concentrates in PAS-plasma, prepared by the OrbiSac or TACSI automated system.

    PubMed

    Plaza, E M; Céspedes, P; Fernández, H; Sánchez-Guiu, M I; Egea, J M; Vicente, V; Lozano, M L; Rivera, J

    2014-01-01

    Buffy-coat (BC)-derived platelet concentrates (PCs) are the predominant product for platelet transfusion in many countries. Two automated systems, OrbiSac and TACSI, have been introduced in blood centres to prepare these PCs, as an alternative to the manual method. We compared the in vitro quality of PCs prepared by both methods during standard storage. Twenty primary BC pools were split into two parts, which were processed with OrbiSac and TACSI system to obtain OrbiSac PCs (O-PCs) and TACSI PCs (T-PCs), respectively. On days 1, 5 and 7 of standard storage, samples were taken and the following analysed: cell count, metabolic variables, platelet function and content of activation and proinflammatory substances. Both the OrbiSac and TACSI systems produced PCs that meet the standards for platelet products in terms of platelet and leucocyte content. In vitro evaluation pointed to the similar preservation of platelet metabolism (pH, glucose, bicarbonate and lactate) in O-PCs and T-PCs. Moreover, there were no significant differences between O-PCs and T-PCs as regards the hypotonic shock response or in the platelet aggregation profile. The OrbiSac system caused greater platelet activation, which resulted in higher concentrations of sCD62P, RANTES and sCD40L on the day the PCs were prepared. The systems OrbiSac and TACSI can be used to produce buffy-coat-derived PCs whose cell content, platelet function and metabolism are similar during standard storage. However, the preparation with the OrbiSac system induces a transient increase in platelet activation and release of proinflammatory substances. © 2013 International Society of Blood Transfusion.

  6. The Bone Regeneration Using Bone Marrow Stromal Cells with Moderate Concentration Platelet-Rich Plasma in Femoral Segmental Defect of Rats

    PubMed Central

    Yamakawa, Junichi; Hashimoto, Junichi; Takano, Mitsuo; Takagi, Michiaki

    2017-01-01

    Background: Platelet-rich plasma (PRP) can provide an assortment of growth factors, but how PRP effects bone regeneration is still unknown. The aim of the study was to explore an optimal method of using PRP and bone marrow stromal cells (BMSCs). Methods: An in vitro experiment was first conducted to determine an appropriate quantity of PRP. BMSCs were cultured with PRP of different concentrations to assess cell proliferation and osteogenic differentiation. Following the in vitro study, a rat femoral segmental defect model was used. Five collagen mixtures consisting of different concentrations of PRP and BMSCs were prepared as follows, i) BMSCs and PRP (platelet 20 x 104/µl), ii) BMSCs and PRP (platelet 100 x 104/µl), iii) BMSCs and PRP (platelet 500 x 104/µl), iv) BMSCs, and v) PRP group (platelet 100 x 104/µl), were used to fill defect. New bone formation was evaluated by soft X-ray and histologic analyses were performed at 2, 4, 6 and 8 weeks postoperatively. Results: The cell proliferation increased PRP concentration-dependently. Cellular alkaline phosphatase activity was higher in moderate concentration than high or low concentration group’s in vitro study. In vivo study, the bone fill percentage of newly formed bone in BMSCs and PRP (platelet 100 x 104/µl) was 46.9% at 8 weeks and increased significantly compared with other groups. Conclusion: BMSCs with moderate level of PRP significantly enhanced bone formation in comparison with BMSCs or PRP transplant in a rat femoral defect model. PMID:28217215

  7. Evaluation of Mirasol pathogen reduction system by artificially contaminating platelet concentrates with Staphylococcus epidermidis: A pilot study from India

    PubMed Central

    Chatterjee, Kabita; Zaman, Shamsuz; Chaurasia, Rahul; Singh, Surinder; Keil, Shawn D.; Tewari, Shalini; Bisht, Akanksha; Agarwal, Nitin; Rout, Diptiranjan; Chand, Subhash; Saha, Kallol

    2016-01-01

    Background and Objectives: This study was conducted to assess the efficacy of Mirasol pathogen reduction system for platelets aimed at preventing bacterial regrowth by spiking buffy coat pooled platelets (BCPP) with clinically relevant load of Staphylococous epidermidis. Materials and Methods: BCPP units were prepared using Teruflex BP-kit with Imugard III-S-PL (Terumo BCT, Tokyo, Japan). Two BCPP units were pooled, of which 40 ml of negative control (NC) was removed. The remaining volume of the platelet unit was inoculated with clinically relevant load of bacteria (total of 30 CFU of S. epidermidis in 1 ml); following this the platelet unit was split into two parts. One part served as positive control (PC) and the other part was subjected to pathogen reduction technique (Mirasol PRT, CaridianBCT Biotechnologies, Lakewood, CO, USA). Bacterial detection was performed using BacT/ALERT system, controls after day 1 and day 7 following inoculation of bacteria and on day 7 for Mirasol-treated unit. Results: Of the 32 treatment cycles, 28 were valid and 4 were invalid. No regrowth was observed in 96.4% (27 of 28) after treatment with Mirasol pathogen reduction system. Of four invalid tests, on two instances the NC showed growth, whereas in other 2 no regrowth was detected in 7th day PC. Bacterial screening of PCs by BacT/ALERT after 24 h of incubation was 28.6%, whereas the effectiveness increased to 100% when incubated for 7 days. Conclusions: Mirasol system was effective in inactivating S. epidermidis when it was deliberately inoculated into BCPP at clinically relevant concentrations. Such systems may significantly improve blood safety by inactivating traditional and emerging transfusion-transmitted pathogens. PMID:27605849

  8. Evaluation of Mirasol pathogen reduction system by artificially contaminating platelet concentrates with Staphylococcus epidermidis: A pilot study from India.

    PubMed

    Chatterjee, Kabita; Zaman, Shamsuz; Chaurasia, Rahul; Singh, Surinder; Keil, Shawn D; Tewari, Shalini; Bisht, Akanksha; Agarwal, Nitin; Rout, Diptiranjan; Chand, Subhash; Saha, Kallol

    2016-01-01

    This study was conducted to assess the efficacy of Mirasol pathogen reduction system for platelets aimed at preventing bacterial regrowth by spiking buffy coat pooled platelets (BCPP) with clinically relevant load of Staphylococous epidermidis. BCPP units were prepared using Teruflex BP-kit with Imugard III-S-PL (Terumo BCT, Tokyo, Japan). Two BCPP units were pooled, of which 40 ml of negative control (NC) was removed. The remaining volume of the platelet unit was inoculated with clinically relevant load of bacteria (total of 30 CFU of S. epidermidis in 1 ml); following this the platelet unit was split into two parts. One part served as positive control (PC) and the other part was subjected to pathogen reduction technique (Mirasol PRT, CaridianBCT Biotechnologies, Lakewood, CO, USA). Bacterial detection was performed using BacT/ALERT system, controls after day 1 and day 7 following inoculation of bacteria and on day 7 for Mirasol-treated unit. Of the 32 treatment cycles, 28 were valid and 4 were invalid. No regrowth was observed in 96.4% (27 of 28) after treatment with Mirasol pathogen reduction system. Of four invalid tests, on two instances the NC showed growth, whereas in other 2 no regrowth was detected in 7(th) day PC. Bacterial screening of PCs by BacT/ALERT after 24 h of incubation was 28.6%, whereas the effectiveness increased to 100% when incubated for 7 days. Mirasol system was effective in inactivating S. epidermidis when it was deliberately inoculated into BCPP at clinically relevant concentrations. Such systems may significantly improve blood safety by inactivating traditional and emerging transfusion-transmitted pathogens.

  9. [Effect of leukocyte contamination on storage of platelet concentrates from buffy coats].

    PubMed

    Klüter, H; Klinger, M; Bauhaus, M; Kirchner, H

    1994-01-01

    We examined the effect of white cell contamination on thrombocytes prepared from pooled buffy coats over a storage period of 8 days. Using this novel technique, a leukocyte depletion filter can be easily integrated during PC preparation. In a paired study (n = 14) eight ABO-identical BC were pooled in a 2-liter PVC bag within 8 h after whole-blood donation, thoroughly mixed and divided into two identical fractions. After soft-spin centrifugation the platelet-rich plasma (PRP) was transferred either (fraction A) using a leukocyte filter (PL 50-HF, Pall) or (fraction B) directly into the storage bag (Pl-732, Baxter), and stored under routine conditions. On days 1, 3, 5, and 8, aliquots of PC were withdrawn for determination of cell count and different biochemical parameters and for morphometric analyses of platelet ultrastructure by electron microscopy. Results showed a lower thrombocyte yield and white cell count (p < 0.01) in fraction A (268 x 10(9) vs. 240 x 10(9); 51.1 x 10(6) vs. 0.04 x 10(6)), whereas no differences between the preparations could be detected by analysis of pH, pCO2, bicarbonate, and in LDH release over the storage period of 8 days. These results were supported in the study on the ultrastructural level where a good morphological integrity of the platelets was observed during the whole storage period in both fractions. In conclusion, storage lesions on platelets due to leukocyte effects are unlikely to occur in PC with white cell counts lower than 10(8)/l.

  10. Growth factor and pro-inflammatory cytokine contents in platelet-rich plasma (PRP), plasma rich in growth factors (PRGF), advanced platelet-rich fibrin (A-PRF), and concentrated growth factors (CGF).

    PubMed

    Masuki, Hideo; Okudera, Toshimitsu; Watanebe, Taisuke; Suzuki, Masashi; Nishiyama, Kazuhiko; Okudera, Hajime; Nakata, Koh; Uematsu, Kohya; Su, Chen-Yao; Kawase, Tomoyuki

    2016-12-01

    The development of platelet-rich fibrin (PRF) drastically simplified the preparation procedure of platelet-concentrated biomaterials, such as platelet-rich plasma (PRP), and facilitated their clinical application. PRF's clinical effectiveness has often been demonstrated in pre-clinical and clinical studies; however, it is still controversial whether growth factors are significantly concentrated in PRF preparations to facilitate wound healing and tissue regeneration. To address this matter, we performed a comparative study of growth factor contents in PRP and its derivatives, such as advanced PRF (A-PRF) and concentrated growth factors (CGF). PRP and its derivatives were prepared from the same peripheral blood samples collected from healthy donors. A-PRF and CGF preparations were homogenized and centrifuged to produce extracts. Platelet and white blood cell counts in A-PRF and CGF preparations were determined by subtracting those counts in red blood cell fractions, supernatant acellular serum fractions, and A-PRF/CGF exudate fractions from those counts of whole blood samples. Concentrations of growth factors (TGF-β1, PDGF-BB, VEGF) and pro-inflammatory cytokines (IL-1β, IL-6) were determined using ELISA kits. Compared to PRP preparations, both A-PRF and CGF extracts contained compatible or higher levels of platelets and platelet-derived growth factors. In a cell proliferation assay, both A-PRF and CGF extracts significantly stimulated the proliferation of human periosteal cells without significant reduction at higher doses. These data clearly demonstrate that both A-PRF and CGF preparations contain significant amounts of growth factors capable of stimulating periosteal cell proliferation, suggesting that A-PRF and CGF preparations function not only as a scaffolding material but also as a reservoir to deliver certain growth factors at the site of application.

  11. Feasibility of optical computerized tomography for measuring the species concentration distribution of flow fields

    NASA Astrophysics Data System (ADS)

    Chen, Yun-yun; Yu, Yang; Chen, Xuan; Zhang, Ying-ying

    2017-08-01

    In this paper, the feasibility of using optical computerized tomography (OCT) methods for measuring the distribution of species concentration for flow fields is analyzed and discussed. First, feasible methods are chosen for two or three objects composed flow fields from the perspective of the measurable principle. Second, both common gas and plasma are chosen as two typical examples for specific analysis and discussion. The results show that the feasibility and applicable range of OCT methods are related to the temperature, pressure, and species composition of the measured flow fields. Finally, the study indicates that OCT methods are more suitable for measuring the distribution of species composition for common gas rather than plasma. In a word, this study could be helpful for extending the applicable range of OCT methods, which are based on the measurement of the refractive index.

  12. Divergent endothelial function but similar platelet microvesicle responses following eccentric and concentric cycling at a similar aerobic power output.

    PubMed

    Rakobowchuk, Mark; Ritter, Ophélie; Wilhelm, Eurico Nestor; Isacco, Laurie; Bouhaddi, Malika; Degano, Bruno; Tordi, Nicolas; Mourot, Laurent

    2017-04-01

    Endothelial function and microvesicle concentration changes after acute bouts of continuous eccentric exercise have not been assessed previously nor compared with concentric exercise at similar aerobic power outputs. This method of training may be useful among some clinical populations, but acute responses are not well described. As such, 12 healthy males completed 2 experimental sessions of either 45 min of eccentric or concentric cycling at a matched aerobic power output below the ventilatory threshold. Brachial artery vascular function was assessed throughout 5 min of forearm ischemia and 3 min thereafter, before and at 5 and 40 min of recovery following each exercise session [flow-mediated dilation (FMD)]. Venous blood samples were acquired before each vascular function assessment. FMD significantly decreased after eccentric cycling by 40 min of recovery (P < 0.05), but was unaltered after concentric exercise. No differences in peak hyperemic blood flow velocity occurred neither between modalities nor at any time point (P > 0.05). Platelet-derived microvesicles increased by ~20% after both exercise modalities (P < 0.05) while endothelial-derived microvesicles were unchanged (P > 0.05). Moderate relationships with cardiac output, a surrogate for shear stress, and norepinephrine were apparent (P < 0.05), but there were no relationships with inflammatory or acute phase proteins. In summary, eccentric endurance exercise induced macrovascular endothelial dysfunction; however, endothelial activation determined by endothelial microvesicles did not occur suggesting that this modality may induce oxidative stress but no significant endothelial damage. In addition, the increase in platelet microvesicle concentrations may induce beneficial microvascular adaptations as suggested by previous research.NEW & NOTEWORTHY Continuous eccentric cycling exercise induces substantial skeletal muscle, tendon, and bone strain providing a potentially beneficial stimulus among clinical

  13. Solvent and concentration effects on the surface characteristics and platelet compatibility of zwitterionic sulfobetaine-terminated self-assembled monolayers.

    PubMed

    Shen, Ching-Hsiung; Lin, Jui-Che

    2013-01-01

    The structural organization of a monolayer influences biological responses as the material makes contact with the bodily fluid. Zwitterionic materials containing the sulfobetaine functionalities have been shown to exhibit protein-repelling characteristics. In this study, the effect of solvent and thiol concentrations on the sulfobetaine-terminated SAM (self-assembled monolayer) is discussed. Four different types of solvents were selected: deionized water, PBS, methanol and ethanol. The total thiol concentration was set at either 2mM or 0.1 mM. X-ray photoelectron analyses indicated that all SAMs demonstrated similar chemical configurations. Reflection adsorption infrared spectroscopy showed that conformation of the SAMs was more organized when prepared from a 0.1 mM solution compared to a 2 mM solution. The contact angle of the SAMs prepared from 2 mM concentration was dependent upon the solvent utilized and was more hydrophobic than the SAMs prepared from 0.1 mM concentration. Moreover, all of these sulfobetaine-terminated SAMs showed a fairly negative zeta potential in PBS at pH 7.4. After contact with blood, these sulfobetaine-terminated SAMs demonstrated distinct platelet reactivity among each other. The highest platelet compatibility was shown on the SAMs prepared in 0.1 mM solution and the one formed in 2 mM ethanolic solution, where they exhibited a more organized conformation and enhanced hydrophilic properties. These properties might be caused by the different hydration layers, which are affected by the assembly conditions on the topmost monolayer. This study demonstrated that optimizing solvent and concentration conditions could control the structural organization of zwitterionic sulfobetaine-terminated SAMs and, consequently, modify biomedical properties.

  14. Quantification of bone mass gain in response to the application of biphasic bioceramics and platelet concentrate in critical-size bone defects.

    PubMed

    Lobo, Sonja Ellen; Wykrota, Francisco Henrique Lanna; Oliveira, Ana Carolina Marques Barbosa; Kerkis, Irina; Mahecha, Germán Bohorquez; Alves, Humberto José

    2009-05-01

    Biphasic bioceramics have been widely indicated for bone reconstruction; however, the real gain in bone mass due to the presence of such biomaterials has not been established yet nor the advantages of its association with platelet concentrate. This study aims at quantifying the volume of bone matrix, osteoblasts, osteocytes, blood vessels and adipose tissue after the application of a biphasic bioceramics composed of 65% hydroxyapatite and 35% beta-tricalcium phosphate. Critical-size bone defects were produced in rabbit femora and reconstructed with bioceramics only, with bioceramics combined with platelet concentrate, with platelet concentrate alone, and with no treatment (blood clot). The quantitative evaluation was performed on histological sections using histomorphometry. Our data provide original evidence that consolidates the indication of bioceramics for clinical bone loss reconstruction. The application of biphasic bioceramics alone led to major bone mass gain and was followed by its association with platelet concentrate. On the other hand, platelet concentrate can contribute to the augmentation and maintenance of the adipose tissue, representing a new field for future applications in plastic surgery.

  15. Filtration through a polyester white cell-reduction filter of plasma-poor platelet concentrates prepared with an acetate-containing additive solution.

    PubMed

    Shimizu, T; Mizuno, S; Yamaguchi, H; Kamiya, T; Kokubo, Y

    1993-09-01

    It is of practical importance to known whether the adsorption of platelets and contaminating white cells (WBCs) by the WBC-reduction filter is altered when platelet concentrates (PCs) are prepared in a plasma-poor condition with an acetate-containing additive solution (Seto sol). Plasma-poor PCs with 11-percent residual plasma were prepared from apheresis platelet-rich plasma by using a sterile docking device with steam-sterilized Seto sol. Seto sol contains 115 mM (115 mmol/L) NaCl, 4 mM (4 mmol/L) KCl, 3 mM (3 mmol/L) MgCl2, 10 mM (10 mmol/L) Na3PO4, 15 mM (15 mmol/L) acetate, 3 mM (3 mmol/L) Na3 citrate, and 10 mM (10 mmol/L) glucose (pH 7.1). The solution was steam-sterilized under nitrogen gas. On Days 1 and 5, pooled Seto sol PCs (2.4 x 10(11) platelets) were filtered with a polyester filter at a flow rate of 10 mL per minute. The WBC-removal rate was over 99.9 percent with a platelet recovery of 88 percent following Day 1 filtration. These values were very similar to those of plasma PCs, and 84-percent recovery was achieved following Day 5 filtration. However, when 1 unit of Seto sol PCs with half the number of platelets was filtered with the polyester filter, platelet recovery was about 16 to 17 percent less than that of plasma PCs. Platelet quality was maintained if pooled Seto sol PCs were filtered on Day 1 and stored for over 4 days. Filtration did not alter platelet function in 1-day-old or 5-day-old Seto sol PCs.(ABSTRACT TRUNCATED AT 250 WORDS)

  16. Adenosine diphosphate-induced aggregation of human platelets in flow through tubes. II. Effect of shear rate, donor sex, and ADP concentration.

    PubMed Central

    Bell, D N; Spain, S; Goldsmith, H L

    1989-01-01

    The effect of shear rate on the adenosine diphosphate-induced aggregation of human platelets in Poiseuille flow was studied using the method described in part I (Bell, D.N., S. Spain, and H.L. Goldsmith. 1989. Biophys. J. 56:817-828). The rate and extent of aggregation in citrated platelet-rich plasma were measured over a range of mean transit time from 0.2 to 8.6 s and mean tube shear rate, G, from 41.9 to 1,920 s-1. At 0.2 microM ADP, changes in the single platelet concentration with time suggest that more than one type of platelet-platelet bond mediates platelet aggregation at physiological shear rates. At low G, a high initial rate of aggregation reflects the formation of a weak bond of high affinity, the strength of which diminishes with time. Here, the fraction of collisions yielding stable doublets, the collision efficiency, reached a maximum of 26%. The collision efficiency decreased with increasing G and was accompanied by a progressive delay in the onset of aggregation. However, the gradual expression of a more shear rate-resistant bond at high shear rates and long mean transit times produced a subsequent increase in collision efficiency and a corresponding increase in the rate of aggregation. Although the collision efficiencies here were less than 1%, the high collision frequencies were able to sustain a high rate of aggregation. At 0.2 microM ADP, aggregate size generally decreased with increasing G. At 1.0 microM ADP, aggregate size was still limited at high shear rates even though the rate of single platelet aggregation was much higher than at 0.2 microM ADP. Platelet aggregation was greater for female than for male donors, an effect related to differences in the hematocrit of donors before preparing platelet-rich plasma. PMID:2605299

  17. The role of in process qualification in quality improvement of the haemonetics MCS plus leucodepleted platelet concentrate.

    PubMed

    Seghatchian, J; Beard, M; Krailadsiri, P

    2000-06-01

    With the implementation of universal leucodepletion in UK all leucodepletion processes have gone through a standard process qualification and quality improvement. The Haemonetics MCS system is a well established automated platelet collection system for the production of double dose leucoreduced platelet concentrate (WBC approximately 70x10(6)/dose). Recently an automated post collection filtration harness system has been introduced (MCS plus LDP) in which platelets are filtered, using an in-line PALL polyester filter (LRFH6 PALL) to reduce the WBC level to below 5x10(6) WBC/dose. This system passed our Phase I evaluation process based on 20-40 runs. However, some changes in the final volume of the products were needed to conform to national guidelines. Large scale trials using the new volume adjusted protocol revealed occasional failure in the leucocyte content. Therefore, 100% testing had to be implemented on all products. A national evaluation was carried out to determine whether changing the filter to a more efficacious one, the LRFXL (PALL) or slowing the filtration flow rate can influence the overall outcome. To reduce donor variability, known donor population were used with identical apheresis conditions. A more consistent and systematic drop in leucocyte content was observed by reducing the flow rate whereas a similar failure (i.e. 1-3%) rate was found both in controls and LRFXL when using the standard head pressure, which is recommended by the manufacturer. A similar failure rate was found using three different low leucocyte counting technologies (Nageotte, flow cytometry and Imagn 2000). It is recommended that a process qualification/validation program should be implemented when even a small modification in the collection system is introduced.

  18. Antiretroviral concentrations in small hair samples as a feasible marker of adherence in rural Kenya

    PubMed Central

    Hickey, Matthew D; Salmen, Charles R; Tessler, Robert A; Omollo, Dan; Bacchetti, Peter; Magerenge, Richard; Mattah, Brian; Salmen, Marcus R; Zoughbie, Daniel; Fiorella, Kathryn J; Geng, Elvin; Njoroge, Betty; Jin, Chengshi; Huang, Yong; Bukusi, Elizabeth A; Cohen, Craig R; Gandhi, Monica

    2014-01-01

    Antiretroviral hair levels objectively quantify drug exposure over time and predict virologic responses. We assessed the acceptability and feasibility of collecting small hair samples in a rural Kenyan cohort. 95% of participants (354/373) donated hair. Although median self-reported adherence was 100% (IQR 96–100%), a wide range of hair concentrations likely indicates overestimation of self-reported adherence and the advantages of a pharmacologic adherence measure. Higher nevirapine (NVP) hair concentrations observed in women and older adults require further study to unravel behavioral versus pharmacokinetic contributors. In resource-limited settings, hair antiretroviral levels may serve as a low-cost quantitative biomarker of adherence. PMID:24694932

  19. OLAND is feasible to treat sewage-like nitrogen concentrations at low hydraulic residence times.

    PubMed

    De Clippeleir, Haydée; Yan, Xungang; Verstraete, Willy; Vlaeminck, Siegfried Elias

    2011-05-01

    Energy-positive sewage treatment can, in principle, be obtained by maximizing energy recovery from concentrated organics and by minimizing energy consumption for concentration and residual nitrogen removal in the main stream. To test the feasibility of the latter, sewage-like nitrogen influent concentrations were treated with oxygen-limited autotrophic nitrification/denitrification (OLAND) in a lab-scale rotating biological contactor at 25°C. At influent ammonium concentrations of 66 and 29 mg N L(-1) and a volumetric loading rate of 840 mg N L(-1) day(-1) yielding hydraulic residence times (HRT) of 2.0 and 1.0 h, respectively, relatively high nitrogen removal rates of 444 and 383 mg N L(-1) day(-1) were obtained, respectively. At low nitrogen levels, adapted nitritation and anammox communities were established. The decrease in nitrogen removal was due to decreased anammox and increased nitratation, with Nitrospira representing 6% of the biofilm. The latter likely occurred given the absence of dissolved oxygen (DO) control, since decreasing the DO concentration from 1.4 to 1.2 mg O(2) L(-1) decreased nitratation by 35% and increased anammox by 32%. Provided a sufficient suppression of nitratation, this study showed the feasibility of OLAND to treat low nitrogen levels at low HRT, a prerequisite to energy-positive sewage treatment.

  20. Measurement of betamethasone concentration in maternal serum treated for fetal lung maturity; Is it feasible?

    PubMed

    Salim, Raed; Suleiman, Abeer; Colodner, Raul; Nachum, Zohar; Goldstein, Lee H; Shalev, Eliezer

    2016-02-10

    The association between maternal serum concentration of betamethasone given for fetal lung maturity and perinatal outcome has not been investigated. This may be due to an absence of a reliable method for measuring serum betamethasone concentrations. We aimed in the current study to assess the feasibility of a specific ELISA kit to measure the concentrations of betamethasone in maternal serum and to examine the trend of sequential measurements after a course of betamethasone for fetal lung maturity. Pregnant women at risk for preterm birth who received betamethasone between 24 and 34 weeks of gestation were prospectively included. Serum concentrations were determined before administering betamethasone (baseline), and 36 hours, 48 hours, 72 hours, and 5 to 7 days after the 1(st) dose. Betamethasone concentration in samples was determined using Corticosteroid ELISA kit. The Friedman test was used to test whether there were significant differences between the measurements. Five singleton pregnancies were included. Using the ELISA kit, betamethasone concentration in maternal serum samples was obtained for all women. Among the five measurements performed, the concentration was highest at 36 hours after the 1(st) dose and close to baseline at the 5(th) measurement performed after 5 to 7 days (p < 0.05). Serum concentration varied at each time point between the five women but similar trend was observed. Betamethasone concentration is measurable in the serum of pregnant women with this ELISA kit.

  1. High plasma fibrinogen concentration and platelet count unfavorably impact survival in non-small cell lung cancer patients with brain metastases.

    PubMed

    Zhu, Jian-Fei; Cai, Ling; Zhang, Xue-Wen; Wen, Yin-Sheng; Su, Xiao-Dong; Rong, Tie-Hua; Zhang, Lan-Jun

    2014-02-01

    High expression of fibrinogen and platelets are often observed in non-small cell lung cancer (NSCLC) patients with local regional or distant metastasis. However, the role of these factors remains unclear. The aims of this study were to evaluate the prognostic significance of plasma fibrinogen concentration and platelet count, as well as to determine the overall survival of NSCLC patients with brain metastases. A total of 275 NSCLC patients with brain metastasis were enrolled into this study. Univariate analysis showed that high plasma fibrinogen concentration was associated with age≥65 years (P = 0.011), smoking status (P = 0.009), intracranial symptoms (P = 0.022), clinical T category (P = 0.010), clinical N category (P = 0.003), increased partial thromboplastin time (P < 0.001), and platelet count (P < 0.001). Patients with low plasma fibrinogen concentration demonstrated longer overall survival compared with those with high plasma fibrinogen concentration (median, 17.3 months versus 11.1 months; P≤0.001). A similar result was observed for platelet counts (median, 16.3 months versus 11.4 months; P = 0.004). Multivariate analysis showed that both plasma fibrinogen concentration and platelet count were independent prognostic factors for NSCLC with brain metastases (R2 = 1.698, P < 0.001 and R2 = 1.699, P < 0.001, respectively). Our results suggest that high plasma fibrinogen concentration and platelet count indicate poor prognosis for NSCLC patients with brain metastases. Thus, these two biomarkers might be independent prognostic predictors for this subgroup of NSCLC patients.

  2. Platelet activation, adhesion, inflammation, and aggregation potential are altered in the presence of electronic cigarette extracts of variable nicotine concentrations.

    PubMed

    Hom, Sarah; Chen, Li; Wang, Tony; Ghebrehiwet, Berhane; Yin, Wei; Rubenstein, David A

    2016-11-01

    Tobacco smoke extracts prepared from both mainstream and sidestream smoking have been associated with heightened platelet activation, aggregation, adhesion, and inflammation. Conversely, it has been shown that pure nicotine inhibits similar platelet functions. In this work, we 1) evaluated the effects of e-cigarette extracts on platelet activities and 2) elucidated the differences between the nicotine-dependent and non-nicotine dependent (e.g. fine particulate matter or toxic compounds) effects of tobacco and e-cigarette products on platelet activities. To accomplish these goals, platelets from healthy volunteers (n = 50) were exposed to tobacco smoke extracts, e-cigarette vapor extracts, and pure nicotine and changes in platelet activation, adhesion, aggregation, and inflammation were evaluated, using optical aggregation, flow cytometry, and ELISA methods. Interestingly, the exposure of platelets to e-vapor extracts induced a significant up-regulation in the expression of the pro-inflammatory gC1qR and cC1qR and induced a marked increase in the deposition of C3b as compared with traditional tobacco smoke extracts. Similarly, platelet activation, as measured by a prothrombinase based assay, and platelet aggregation were also significantly enhanced after exposure to e-vapor extracts. Finally, platelet adhesion potential toward fibrinogen, von Willebrand factor, and other platelets was also enhanced after exposure to e-cigarette vapor extracts. In the presence of pure nicotine, platelet functions were observed to be inhibited, which further suggests that other constituents of tobacco smoke and electronic vapor can antagonize platelet functions, however, the presence of nicotine in extracts somewhat perpetuated the platelet functional changes in a dose-dependent manner.

  3. [Pathogen inactivation of platelets: organization consequences for platelet transfusion].

    PubMed

    Chavarin, P; DePutter, C; Boussoulade, F; Acquart, S; Vidal, M; Argaud, C; Fabrigli, P; Garraud, O

    2011-08-01

    In the past few years, pathogen reduction technologies for labile blood products have been part of the enhancement of global transfusion safety regarding residual risks of transmitting infectious pathogens. Having carried out a feasibility study for the implementation of pathogen inactivation of platelet concentrates by means of the amotosalen/HCl/UVA (Intercept™) technology, and participated to a reinforced haemovigilance study, we took the opportunity to analyze the organization consequences for platelet concentrates inventory and distribution. This impact study first indicated that those novel needs forced the blood donation service, as well as the labile blood product preparation laboratory, to review and improve practices; secondly, it showed that the routine implementation has little (no major) consequence in the overall organization, independently of the economic consequences (not covered here).

  4. Ultrasound-based measurement of molecular marker concentration in large blood vessels: a feasibility study.

    PubMed

    Wang, Shiying; Mauldin, F William; Klibanov, Alexander L; Hossack, John A

    2015-01-01

    Ultrasound molecular imaging has demonstrated efficacy in pre-clinical studies for cancer and cardiovascular inflammation. However, these techniques often require lengthy protocols because of waiting periods or additional control microbubble injections. Moreover, they are not capable of quantifying molecular marker concentration in human tissue environments that exhibit variable attenuation and propagation path lengths. Our group recently investigated a modulated acoustic radiation force-based imaging sequence, which was found to detect targeted adhesion independent of control measurements. In the present study, this sequence was tested against various experimental parameters to determine its feasibility for quantitative measurements of molecular marker concentration. Results indicated that measurements obtained from the sequence (residual-to-saturation ratio, Rresid) were independent of acoustic pressure and attenuation (p > 0.13, n = 10) when acoustic pressures were sufficiently low. The Rresid parameter exhibited a linear relationship with measured molecular marker concentration (R(2) > 0.94). Consequently, feasibility was illustrated in vitro, for quantification of molecular marker concentration in large vessels using a modulated acoustic radiation force-based sequence. Moreover, these measurements were independent of absolute acoustic reflection amplitude and used short imaging protocols (3 min) without control measurements. Copyright © 2015 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

  5. Evaluation of the 3D BacT/ALERT automated culture system for the detection of microbial contamination of platelet concentrates.

    PubMed

    McDonald, C P; Rogers, A; Cox, M; Smith, R; Roy, A; Robbins, S; Hartley, S; Barbara, J A J; Rothenberg, S; Stutzman, L; Widders, G

    2002-10-01

    Bacterial transmission remains the major component of morbidity and mortality associated with transfusion-transmitted infections. Platelet concentrates are the most common cause of bacterial transmission. The BacT/ALERT 3D automated blood culture system has the potential to screen platelet concentrates for the presence of bacteria. Evaluation of this system was performed by spiking day 2 apheresis platelet units with individual bacterial isolates at final concentrations of 10 and 100 colony-forming units (cfu) mL-1. Fifteen organisms were used which had been cited in platelet transmission and monitoring studies. BacT/ALERT times to detection were compared with thioglycollate broth cultures, and the performance of five types of BacT/ALERT culture bottles was evaluated. Sampling was performed immediately after the inoculation of the units, and 10 replicates were performed per organism concentration for each of the five types of BacT/ALERT bottles. The mean times for the detection of these 15 organisms by BacT/ALERT, with the exception of Propionibacterium acnes, ranged from 9.1 to 48.1 h (all 10 replicates were positive). In comparison, the time range found using thioglycollate was 12.0-32.3 h (all 10 replicates were positive). P. acnes' BacT/ALERT mean detection times ranged from 89.0 to 177.6 h compared with 75.6-86.4 h for the thioglycollate broth. BacT/ALERT, with the exception of P. acnes, which has dubious clinical significance, gave equivalent or shorter detection times when compared with the thioglycollate broth system. The BacT/ALERT system detected a range of organisms at levels of 10 and 100 cfu mL-1. This study validates the BacT/ALERT microbial detection system for screening platelets. Currently, the system is the only practically viable option available for routinely screening platelet concentrates to prevent bacterial transmission.

  6. The expression and concentration of CD40 ligand in normal pregnancy, preeclampsia, and hemolytic anemia, elevated liver enzymes and low platelet count (HELLP) syndrome.

    PubMed

    Azzam, Hanan A G; Abousamra, Nashwa K; Goda, Hossam; El-Shouky, Reda; El-Gilany, Abdel-Hady

    2013-01-01

    Preeclampsia has been associated with increased platelet activation detected before disease onset. Inappropriate activation of platelets may be involved in pathogenesis in preeclampsia by promoting coagulation and thrombosis and also as a mediator of inflammation. The exaggerated platelet activation and inflammation leading to endothelial damage in preeclampsia can be explained by the CD40-CD40 ligand (CD40L) system. Expression of CD40L on platelets was determined by whole-blood flow cytometry, and serum levels of soluble CD40L (sCD40L) were measured by enzyme-linked immunosorbent assay in 11 women with mild preeclampsia, 11 women with severe preeclampsia, and six women with hemolytic anemia, elevated liver enzymes and low platelet count (HELLP) syndrome compared with 13 normotensive pregnant women as a control group. The platelet surface expression of CD40L was significantly higher in women with mild and severe preeclampsia and HELLP compared with normal pregnancy group (P = 0.001; P ≤ 0.001; P = 0.003, respectively), with no significant difference being found between women with mild preeclampsia compared with HELLP and severe preeclampsia compared with HELLP (P = 0.2; P = 0.8, respectively). The serum concentration of sCD40L was significantly higher in women with mild and severe preeclampsia and HELLP compared with the normal pregnancy group (P = 0.001; P ≤ 0.001; P = 0.022, respectively), with no significant difference being found between women with mild compared with severe preeclampsia or HELLP and severe preeclampsia compared with HELLP (P = 0.7; P = 0.6; P = 0.6, respectively). In conclusion, the higher expression and concentration of CD40L in women with preeclampsia and HELLP syndrome compared with normal pregnant women may indicate an exaggerated activation of platelets and endothelial cells in the disorder.

  7. Resting and ADP-evoked changes in cytosolic free sodium concentration in human platelets loaded with the indicator SBFI.

    PubMed Central

    Sage, S O; Rink, T J; Mahaut-Smith, M P

    1991-01-01

    1. Cytosolic free Na+ concentration, [Na+]i, was investigated in human platelets loaded with the fluorescent indicator SBFI (sodium-binding benzofuran isophthalate). 2. SBFI fluorescence from platelet suspensions was measured at excitation wavelengths of 340 and 385 nm and the 340/385 nm fluorescence ratio was calibrated in terms of [Na+]i in situ. [Na+]i was set to known values by resuspending cells in media with various [Na+], in the presence of the Na(+)-K+ ionophore, gramicidin. 3. Basal free [Na+]i was 5.5 +/- 0.3 mM (n = 50). This is considerably lower than estimates of total platelet Na+, suggesting that much intracellular Na+ is sequestered or bound. 4. ADP (40 microM) evoked a rise in [Na+]i from 6.4 +/- 0.7 to 18.3 +/- 1.1 mM (n = 8). The ADP-evoked rise in [Na+]i was abolished when external Na+ was replaced with N-methyl-D-glucamine. This indicates that the rise in [Na+]i was due to Na+ entry. 5. In platelets loaded with the fluorescent pH indicator, BCECF, 40 microM-ADP was shown to evoke a fall in cytosolic pH (pHi) from 7.21 +/- 0.03 to 7.12 +/- 0.03 (n = 10). Three minutes after ADP addition pHi had only recovered to 7.15 +/- 0.03. The recovery was dependent on external Na+, suggesting it was mediated by Na(+)-H+ exchange. However, this would only account for an increase in [Na+]i of approximately 0.5 mM, indicating most of the ADP-evoked Na+ entry occurred by other mechanisms. 6. Stopped-flow fluorimetry showed that the ADP-evoked rise in [Na+]i commenced without measurable delay and peaked within 1 s. The initial kinetics were thus similar to those reported for ADP-evoked rises in [Ca2+]i. 7. Cell-attached patch-clamp recordings showed that ADP evoked single-channel inward currents when included in the pipette-filling solution. The currents were similar whether Ca2+ was present or absent from the pipette. The slope conductance was 11 pS in the presence of external Ca2+ and 10 pS in its absence. Current-voltage relationships were similar and the

  8. Platelet-collagen adhesion enhances platelet aggregation induced by binding of VWF to platelets

    SciTech Connect

    Laduca, F.M.; Bell, W.R.; Bettigole, R.E. State Univ. of New York, Buffalo )

    1987-11-01

    Ristocetin-induced platelet aggregation (RIPA) was evaluated in the presence of platelet-collagen adhesion. RIPA of normal donor platelet-rich plasma (PRP) demonstrated a primary wave of aggregation mediated by the binding of von Willebrand factor (VWF) to platelets and a secondary aggregation wave, due to a platelet-release reaction, initiated by VWF-platelet binding and inhibitable by acetylsalicylic acid (ASA). An enhanced RIPA was observed in PRP samples to which collagen had been previously added. These subthreshold concentrations of collagen, which by themselves were insufficient to induce aggregation, caused measurable platelet-collagen adhesion. Subthreshold collagen did not cause microplatelet aggregation, platelet release of ({sup 3}H)serotonin, or alter the dose-responsive binding of {sup 125}I-labeled VWF to platelets, which occurred with increasing ristocetin concentrations. However, ASA inhibition of the platelet release reaction prevented collagen-enhanced RIPA. These results demonstrate that platelet-collagen adhesion altered the platelet-release reaction induced by the binding of VWF to platelets causing a platelet-release reaction at a level of VWF-platelet binding not normally initiating a secondary aggregation. These findings suggest that platelet-collagen adhesion enhances platelet function mediated by VWF.

  9. Gadofosveset-Based Biomarker of Tissue Albumin Concentration: Technical Validation in Vitro and Feasibility in Vivo

    PubMed Central

    Richardson, Owen C; Bane, Octavia; Scott, Marietta LJ; Tanner, Steven F; Waterton, John C; Sourbron, Steven P; Carroll, Timothy J; Buckley, David L

    2015-01-01

    Purpose There is currently no adequate method of mapping physiologic and pathophysiologic tissue albumin concentrations in human subjects. The objective of this study was to devise and evaluate a biomarker of regional albumin concentration using gadofosveset-enhanced MRI. Theory and Methods A binding and relaxation model was devised and evaluated in vitro in solutions of albumin at 3.0 Tesla (T) and 4.7T. The method was evaluated in the heart in seven volunteers at 3.0T. Results MRI-derived estimates of albumin concentration were in good agreement with true values over the range 0.1–1.0 mM (Pearson correlation coefficients of 0.85 and 0.88 for 3.0T and 4.7T, respectively). The mean calculated albumin concentration in the myocardium for the volunteers was 0.02 mM (range, 0.01–0.03 mM). Conclusion Accurate estimates of albumin concentration in vitro suggest this may be a viable noninvasive alternative to existing techniques. In the myocardium the MRI-derived estimates of albumin concentration indicate the practical feasibility of the technique but were below expected values. Gadofosveset-enhanced MR relaxometry has potential in providing biomarkers of regional albumin concentration; further evaluation is required before it can be used reliably in vivo. Magn Reson Med 73:244–253, 2015. © 2014 Wiley Periodicals, Inc. PMID:24515975

  10. A clinically-feasible protocol for using human platelet lysate and mesenchymal stem cells in regenerative therapies.

    PubMed

    Warnke, Patrick H; Humpe, Andreas; Strunk, Dirk; Stephens, Sebastien; Warnke, Frauke; Wiltfang, Joerg; Schallmoser, Katharina; Alamein, Mohammad; Bourke, Robert; Heiner, Peter; Liu, Qin

    2013-03-01

    The transplantation of human stem cells seeded on biomaterials holds promise for many clinical applications in cranio-maxillo-facial tissue engineering and regenerative medicine. However, stem cell propagation necessary to produce sufficient cell numbers currently utilizes fetal calf serum (FCS) as a growth supplement which may subsequently transmit animal pathogens. Human platelet lysate (HPL) could potentially be utilized to produce clinical-grade stem cell-loaded biomaterials as an appropriate FCS substitute that is in line with clinically-applicable practice. The goal of this study was to investigate whether HPL can be successfully used to propagate human mesenchymal stem cells (HMSCs) seeded on clinically-approved collagen materials under clinically-applicable conditions using FCS as a control. HMSCs were isolated from bone marrow and cultured in the presence of 10% FCS or 10% HPL. Characterization of HMSCs was performed by flow cytometry and through osteogenic and adipogenic differentiation assays. Proliferative capacity of HMSCs on both matrices was investigated by mitochondrial dehydrogenase assays (WST) and tissue coverage scanning electron microscopy (SEM). The isolated HMSC differentiated into osteogenic and adipogenic cells authenticating the multipotentiality of the HMSCs. WST tests and the SEM images demonstrated that HPL was generally superior to FCS in promoting growth of seeded HMSCs. For all other tests HPL supported HMSCs at least equal to FCS. In conclusion, HPL is an effective growth factor to allow expansion of clinical-grade HMSCs on clinically-approved biomaterials for maxillofacial and oral implantology applications.

  11. Evaluation of the effect of calcium gluconate and bovine thrombin on the temporal release of transforming growth factor beta 1 and platelet-derived growth factor isoform BB from feline platelet concentrates.

    PubMed

    Silva, Raul F; Alvarez, María E; Ríos, Diana L; López, Catalina; Carmona, Jorge U; Rezende, Cleuza Mf

    2012-11-06

    There are not reported regarding the protocols for obtaining platelet concentrates (PC) in cats for medical purposes. The objectives of this study were: 1) to describe a manual method for producing two kinds of PC in cats (PC-A and PC-B), 2) to describe the cellular population of the PC, 3) to measure and compare the effect of calcium gluconate (CG) and bovine thrombin (BT) on the temporal release of transforming growth factor beta 1 (TGF-β1) and platelet-derived growth factor type BB (PDGF-BB) at 3 and 12 hours post-activation and 4) to establish correlations between the cellular population of both PCs and the concentration of growth factors (GF). Blood samples were taken from 16 cats for complete blood count, plasma collection and PC preparation. The PC were arbitrarily divided into two fractions, specifically, PC-A (lower fraction) and PC-B (upper fraction). The platelet counts were significantly different (P<0.05) between the PC and whole blood but not between the PC fractions. The TGF-β1 concentration efficiencies for PC-A and PC-B activated with CG were 42.86% and 46.54%, and activated with BT were 42.88% and 54.64%, respectively. The PDGF-BB concentration efficiencies for PC-A and PC-B activated with CG were 61.36% and 60.61%, and activated with BT were 65.64% and 72.12%, respectively. The temporal release of GFs showed no statistically significant difference (P>0.05) between the activating substances at the time or for any PC fraction. Whatever the activation means, these preparations of cat PC provide significant concentrations of platelets and GFs for possible clinical or experimental use.

  12. Evaluation of the effect of calcium gluconate and bovine thrombin on the temporal release of transforming growth factor beta 1 and platelet-derived growth factor isoform BB from feline platelet concentrates

    PubMed Central

    2012-01-01

    Background There are not reported regarding the protocols for obtaining platelet concentrates (PC) in cats for medical purposes. The objectives of this study were: 1) to describe a manual method for producing two kinds of PC in cats (PC-A and PC-B), 2) to describe the cellular population of the PC, 3) to measure and compare the effect of calcium gluconate (CG) and bovine thrombin (BT) on the temporal release of transforming growth factor beta 1 (TGF-β1) and platelet-derived growth factor type BB (PDGF-BB) at 3 and 12 hours post-activation and 4) to establish correlations between the cellular population of both PCs and the concentration of growth factors (GF). Blood samples were taken from 16 cats for complete blood count, plasma collection and PC preparation. The PC were arbitrarily divided into two fractions, specifically, PC-A (lower fraction) and PC-B (upper fraction). Results The platelet counts were significantly different (P<0.05) between the PC and whole blood but not between the PC fractions. The TGF-β1 concentration efficiencies for PC-A and PC-B activated with CG were 42.86% and 46.54%, and activated with BT were 42.88% and 54.64%, respectively. The PDGF-BB concentration efficiencies for PC-A and PC-B activated with CG were 61.36% and 60.61%, and activated with BT were 65.64% and 72.12%, respectively. The temporal release of GFs showed no statistically significant difference (P>0.05) between the activating substances at the time or for any PC fraction. Conclusions Whatever the activation means, these preparations of cat PC provide significant concentrations of platelets and GFs for possible clinical or experimental use. PMID:23131192

  13. Management of an endo perio lesion in a maxillary canine using platelet-rich plasma concentrate and an alloplastic bone substitute

    PubMed Central

    Singh, Sangeeta

    2009-01-01

    To evaluate the efficacy of platelet-rich plasma concentrate in the management of a cirumferential, infrabony defect associated with an endoperio lesion in a maxillary canine. A 45 year-old male patient with an endoperio lesion in the left maxillary canine was initially treated with endodontic therapy. Following the endodontic treatment, the circumferential, infrabony defect was treated using platelet-rich plasma and an alloplastic bone substitute. At the end of three months, there was a gain in the clinical attachment level and reduction in probing depth. Radiographic evidence showed that there was significant bony fill. The results were maintained at the time of recall nine months later. PMID:20407658

  14. Detection of enteric viruses in activated sludge by feasible concentration methods.

    PubMed

    Prado, Tatiana; Gaspar, Ana Maria Coimbra; Miagostovich, Marize Pereira

    2014-01-01

    Human enteric viruses are responsible to cause several diseases, including gastroenteritis and hepatitis, and can be present in high amounts in sewage sludge. This study compared virus recovery efficiency of two feasible concentration methods used for detecting human adenovirus (HAdV), rotavirus species A (RV-A), norovirus genogroup II (NoV GII) and hepatitis A virus (HAV) in sewage sludge from an activated sludge process. Twelve sewage sludge samples were collected bi-monthly from January to July, 2011. Ultracentrifugation was compared with a simplified protocol based on beef extract elution for recovering enteric viruses. Viruses were quantified by quantitative real-time PCR assays and virus recovery efficiency and limits of detection were determined. Methods showed mean recovery rates lower than 7.5%, presenting critical limits of detection (higher than 10(2) - 10(3) genome copies - GC L(-1) for all viruses analyzed). Nevertheless, HAdV were detected in 90% of the analyzed sewage sludge samples (range: 1.8 × 10(4) to 1.1 × 10(5) GC L(-1)), followed by RV-A and NoV (both in 50%) and HAV (8%). Results suggesting that activated sludge is contaminated with high viral loads and HAdV are widely disseminated in these samples. The low virus recovery rates achieved, especially for HAV, indicate that other feasible concentration methods could be developed to improve virus recovery efficiency in these environmental matrices.

  15. Detection of enteric viruses in activated sludge by feasible concentration methods

    PubMed Central

    Prado, Tatiana; Gaspar, Ana Maria Coimbra; Miagostovich, Marize Pereira

    2014-01-01

    Human enteric viruses are responsible to cause several diseases, including gastroenteritis and hepatitis, and can be present in high amounts in sewage sludge. This study compared virus recovery efficiency of two feasible concentration methods used for detecting human adenovirus (HAdV), rotavirus species A (RV-A), norovirus genogroup II (NoV GII) and hepatitis A virus (HAV) in sewage sludge from an activated sludge process. Twelve sewage sludge samples were collected bi-monthly from January to July, 2011. Ultracentrifugation was compared with a simplified protocol based on beef extract elution for recovering enteric viruses. Viruses were quantified by quantitative real-time PCR assays and virus recovery efficiency and limits of detection were determined. Methods showed mean recovery rates lower than 7.5%, presenting critical limits of detection (higher than 102 – 103 genome copies - GC L−1 for all viruses analyzed). Nevertheless, HAdV were detected in 90% of the analyzed sewage sludge samples (range: 1.8 × 104 to 1.1 × 105 GC L−1), followed by RV-A and NoV (both in 50%) and HAV (8%). Results suggesting that activated sludge is contaminated with high viral loads and HAdV are widely disseminated in these samples. The low virus recovery rates achieved, especially for HAV, indicate that other feasible concentration methods could be developed to improve virus recovery efficiency in these environmental matrices. PMID:24948954

  16. The PFA-100R cannot detect blood group-dependent inhibition of platelet function by eptifibatide or abciximab at therapeutic plasma concentrations.

    PubMed

    Feuring, M; Ruf, A; Schultz, A; Wehling, M

    2010-01-01

    Previous investigations revealed that AB0 blood groups are associated with divergent concentrations of several coagulation factors. Concentrations of von Willebrand factor (vWF) and factor VIII are lower in individuals with blood group 0 compared to subjects with blood group A, B or AB, which might in turn result in a reduced inhibition of platelet aggregation in individuals with blood group 0. The aim of the present in vitro investigation was to elucidate the impact of AB0 blood group-dependent vWF concentrations on eptifibatide and abciximab mediated inhibition of GPIIb/IIIa function. Platelet function was measured with the platelet function analyzer PFA-100(R) at baseline and at increasing concentrations of eptifibatide and abciximab. It was stratified for blood group 0 vs A. If measured with the collagen/ADP cartridge, blood group 0 was associated with a prolonged mean baseline closure time in comparison with blood group A (94.3 +/- 14.6 s vs. 74.6 +/- 9.9 s, p = 0.007) which was paralleled by reduced concentrations of vWF and factor VIII. In contrast, no statistically significant differences in closure times (167.4 +/- 83.9 s vs. 140.1 +/- 99.0 s, p = 0.562) could be found in the presence of eptifibatide (0.1 microg/ml). Higher concentrations of abciximab (1 microg/ml) than those of eptifibatide were needed to increase the closure times in both cartridges of the PFA-100, but at this concentration of abciximab differences in closure times could not be detected most probably due to higher variability at these drug concentrations. The PFA-100(R) is not suitable for monitoring abciximab or eptifibatide within the therapeutic concentration range because the highest concentrations where the PFA-100(R) had measurable closure times of below 300 s is much too low to lead to the necessary platelet inhibition and, consequently, does not resemble the in vivo situation.

  17. Clinical-grade quality platelet-rich plasma releasate (PRP-R/SRGF) from CaCl2 -activated platelet concentrates promoted expansion of mesenchymal stromal cells.

    PubMed

    Borghese, C; Agostini, F; Durante, C; Colombatti, A; Mazzucato, M; Aldinucci, D

    2016-08-01

    The aim of our study was to test a platelet-rich plasma releasate (PRP-R/SRGF) from CaCl2 -activated platelets as a source of growth factors for the expansion of mesenchymal stromal cells (MSCs). PRP-R/SRGF, obtained with a low-cost procedure, is characterized by a reduced variability of growth factor release. PRP-R/SRGF is a clinical-grade quality solution obtained from CaCl2 -activated platelets. Its activity was evaluated by measuring the proliferation, the phenotype, the differentiation potential and the immunosuppressive properties of MSCs derived from bone marrow (BM) and adipose tissue (AT). PRP-R/SRGF was more active than FBS to expand BM- and AT-derived MSCs. PRP-R/SRGF treatment did not affect the expression of typical MSCs surface markers, neither MSCs differentiation potential nor their capability to inhibit activated T-cell proliferation. The clinical-grade PRP-R/SRGF may be used in the clinical setting for the expansion of MSCs. © 2016 International Society of Blood Transfusion.

  18. Do the fibrin architecture and leukocyte content influence the growth factor release of platelet concentrates? An evidence-based answer comparing a pure platelet-rich plasma (P-PRP) gel and a leukocyte- and platelet-rich fibrin (L-PRF).

    PubMed

    Dohan Ehrenfest, David M; Bielecki, Tomasz; Jimbo, Ryo; Barbé, Giovanni; Del Corso, Marco; Inchingolo, Francesco; Sammartino, Gilberto

    2012-06-01

    Platelet concentrates for surgical use are tools of regenerative medicine designed for the local release of platelet growth factors into a surgical or wounded site, in order to stimulate tissue healing or regeneration. Leukocyte content and fibrin architecture are 2 key characteristics of all platelet concentrates and allow to classify these technologies in 4 families, but very little is known about the impact of these 2 parameters on the intrinsic biology of these products. In this demonstration, we highlight some outstanding differences in the growth factor and matrix protein release between 2 families of platelet concentrate: Pure Platelet-Rich Plasma (P-PRP, here the Anitua's PRGF - Preparation Rich in Growth Factors - technique) and Leukocyte- and Platelet-Rich Fibrin (L-PRF, here the Choukroun's method). These 2 families are the extreme opposites in terms of fibrin architecture and leukocyte content. The slow release of 3 key growth factors (Transforming Growth Factor β1 (TGFβ1), Platelet-Derived Growth Factor AB (PDGF-AB) and Vascular Endothelial Growth Factor (VEGF)) and matrix proteins (fibronectin, vitronectin and thrombospondin-1) from the L-PRF and P-PRP gel membranes in culture medium is described and discussed. During 7 days, the L-PRF membranes slowly release significantly larger amounts of all these molecules than the P-PRP gel membranes, and the 2 products display different release patterns. In both platelet concentrates, vitronectin is the sole molecule to be released almost completely after only 4 hours, suggesting that this molecule is not trapped in the fibrin matrix and not produced by the leukocytes. Moreover the P-PRP gel membranes completely dissolve in the culture medium after less than 5 days only, while the L-PRF membranes are still intact after 7 days. This simple demonstration shows that the polymerization and final architecture of the fibrin matrix considerably influence the strength and the growth factor trapping/release potential

  19. Addition of platelet concentrate to dermo-epidermal skin graft in deep burn trauma reduces scarring and need for revision surgeries.

    PubMed

    Prochazka, Vaclav; Klosova, Hana; Stetinsky, Jiri; Gumulec, Jaromir; Vitkova, Katerina; Salounova, Dana; Dvorackova, Jana; Bielnikova, Hana; Klement, Petr; Levakova, Veronika; Ocelka, Tomas; Pavliska, Lubomir; Kovanic, Pavel; Klement, Giannoula Lakka

    2014-06-01

    [corrected] Deep skin burn injuries, especially those on the face, hands, feet, genitalia and perineum represent significant therapeutic challenges. Autologous dermo-epidermal skin grafts (DESG) have become standard of care for treating deep burns. Additionally, human autologous thrombin activated autologous platelet concentrate (APC) has gained acceptance in the setting of wounds. While each of these interventions has been independently shown to accelerate healing, the combination of the two has never been evaluated. We hypothesized that the addition of platelets (source of growth factors and inhibitors necessary for tissue repair) to the DESG (source of progenitor cells and of tissue proteases necessary for spatial and temporal control of growth regulators released from platelets) would create the optimal environment for the reciprocal interaction of cells within the healing tissues. We used clinical examination (digital photography), standardised scales for evaluating pain and scarring, in combination with blood perfusion (laser Doppler imaging), as well as molecular and laboratory analyses. We show for the first time that the combination of APC and DESG leads to earlier relief of pain, and decreased use of analgesics, antipruritics and orthotic devices. Most importantly, this treatment is associated with earlier discharges from hospital and significant cost savings. Our findings indicate that DESG engraftment is facilitated by the local addition of platelets and by systemic thrombocytosis. This local interaction leads to the physiological revascularization at 1-3 months. We observed significant elevation of circulating platelets in early stages of engraftment (1-7 days), which normalized over the subsequent 7 and 90 days.

  20. Platelet concentrates prepared after a 20- to 24-hour hold of the whole blood at 22°C.

    PubMed

    Slichter, Sherrill J; Corson, Jill; Jones, Mary Kay; Christoffel, Todd; Pellham, Esther; Bolgiano, Doug

    2012-09-01

    The Food and Drug Administration (FDA) requires that red blood cells must be refrigerated within 8 hours of whole blood collection. Longer storage of whole blood at 22°C before component preparation would have many advantages. Two methods of holding whole blood for 20 to 24 hours at room temperature were evaluated, refrigerated plates or a 23°C incubator. After extended whole blood storage, platelet (PLT) concentrates were prepared from PLT-rich plasma on Day 1 postdonation, and the PLTs were stored for 6 more days. On Day 7 of PLT storage, blood was drawn from each subject to prepare fresh PLTs. The stored and fresh PLTs were radiolabeled and transfused into their donor. Eleven subjects' whole blood was stored using refrigerated butanediol plates (Compocool, Fresenius), and 10 using an incubator. Poststorage PLT recoveries averaged 47 ± 13% versus 53 ± 11% and survivals averaged 4.6 ± 1.7 days versus 4.7 ± 0.9 days for Compocool versus incubator storage, respectively (p = NS). With all results, poststorage PLT recoveries averaged 75 ± 10% of fresh and survivals 57 ± 13% of fresh; PLT recoveries met FDA guidelines for poststorage PLT viability but not survivals. Seven-day poststorage PLT viability is comparable when whole blood is stored for 22 ± 2 hours at 22°C using either refrigerated plates or an incubator to maintain temperature before preparing PLT concentrates. © 2012 American Association of Blood Banks.

  1. Effect of platelet-rich plasma concentrations on the proliferation of periodontal cells: An in vitro study

    PubMed Central

    Tavassoli-Hojjati, Sara; Sattari, Mandana; Ghasemi, Tayebeh; Ahmadi, Rahil; Mashayekhi, Abbas

    2016-01-01

    Objective: The purpose of this study was to evaluate the effect of different concentrations of platelet-rich plasma (PRP) on the proliferation of undifferentiated periodontal ligament (PDL) fibroblasts. Materials and Methods: The undifferentiated PDL fibroblasts were obtained from two healthy human premolar teeth and cultured in Dulbecco's modified Eagle's medium. Cell wells were divided into five groups. Experimental groups received 0.1%, 5%, or 50% PRP; the positive and negative control groups were cultured in fetal bovine serum (FBS) 12% and in a medium without FBS 12%, respectively. The plates were incubated at 37°C for 1, 2, 3, 4, and 7 days. PDL cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide proliferation assay. Statistical analysis of the data was accomplished using repeated measure ANOVA and Tukey's test. P < 0.05 was considered statistically significant. Results: The 5% PRP had the greatest effect on undifferentiated fibroblast proliferation, which was significant on the 3rd day. There was no significant difference between 0.1% PRP and positive control during the first 3 days. The group with 50% PRP presented significantly lower proliferation, compared to other experimental and control groups. Conclusions: It may be concluded that the growth-stimulating effect of PRP is dose dependent with the best results in low concentrations. PMID:28042260

  2. Platelet interaction with polymerizing fibrin.

    PubMed

    Niewiarowski, S; Regoeczi, E; Stewart, G J; Senyl, A F; Mustard, J F

    1972-03-01

    Interaction of washed pig, rabbit, or human platelets with fibrinogen was studied during its transition to fibrin using photometric, isotopic, and electron microscopic techniques. Untreated fibrinogen and fully polymerized fibrin had no detectable effect on platelets. Fibrinogen, incubated with low concentrations of reptilase or thrombin, formed intermediate products which readily became associated with platelets and caused their aggregation. Neutralization of the thrombin did not prevent this interaction. In the absence of fibrinogen, reptilase did not affect platelets. The interaction of polymerizing fibrin with platelets was accompanied by small losses of platelet constituents (serotonin, adenine nucleotides, platelet factor 4, and lactic dehydrogenase). This loss did not appear to be the result of the platelet release reaction. Inhibitors of the release reaction or of adenosine diphosphate (ADP)-induced aggregation did not prevent the interaction of platelets with polymerizing fibrin. Apyrase or prostaglandin E(1) (PGE(1)) reduced the extent of platelet aggregation by polymerizing fibrin, but the amount of protein associated with platelets was slightly increased. The interaction of polymerizing fibrin with platelets was completely inhibited by ethylenediaminetetraacetate (EDTA) or ethylene glycol bis (beta-aminoethyl ether) N, N,N',N'-tetraacetic acid (EGTA).Fibers formed in solutions of polymerizing fibrin were larger in the presence than in the absence of washed platelets, suggesting that platelets affect fibrin polymerization. The adherence of platelets to polymerizing fibrin may be responsible for the establishment of links between platelets and fibrin in hemostatic plugs and thrombi.

  3. Effects over time of two platelet gel supernatants on growth factor, cytokine and hyaluronan concentrations in normal synovial membrane explants challenged with lipopolysaccharide.

    PubMed

    Ríos, Diana L; López, Catalina; Álvarez, María E; Samudio, Ismael J; Carmona, Jorge U

    2015-06-20

    Platelet-rich plasma (PRP) preparations are a common treatment in osteoarthritis (OA) and inflammatory synovitis. However, there is ambiguity regarding the ideal concentration of leukocytes and platelets in these preparations necessary to induce an adequate anti-inflammatory and anabolic response in joint tissues, such as the synovial membrane. This research aimed to study, in normal synovial membrane explants (SME) challenged with lipopolysaccharide (LPS), the temporal effects (at 48 and 96h) of leukocyte- and platelet-rich gel (L-PRG) and pure platelet-rich gel (P-PRG) supernatants on the production and degradation of platelet associated growth factors (GF) (platelet derived GF isoform BB (PDGF-BB) and transforming growth factor beta-1 (TGF-β1)), pro-inflammatory (tumour necrosis factor alpha (TNF-α)) and anti-inflammatory cytokines (interleukin 4 (IL-4) and IL-1 receptor antagonists (IL-1ra)) and hyaluronan (HA). Synovial membrane explants (SMEs) from 6 horses were challenged with LPS and cultured for 96h with L-PRG and P-PRG supernatants at concentrations of 25 and 50 %, respectively. The SME culture medium was changed every 48h and used for determination by ELISA of PDGF-BB, TGF-β1, TNF-α, IL-4, IL-1ra and HA. These molecules were also determined in synovial fluid from the horses. Both the 25 and 50 % PRG supernatants produced a molecular profile in the culture media unlike that of the SME challenged with LPS only. They presented GF, cytokine and HA concentrations very near to the concentrations of these molecules in normal synovial fluid when compared with the SME control groups (either with LPS or without LPS). However, in comparison with the rest of the SME treated groups, the 25 % L-PRG produced the most IL-1ra, and the 50 % P-PRG induced the sustained production of IL-4 and HA. These in vitro findings suggest that anabolic and anti-inflammatory joint responses depend on the leukocyte and platelet concentration of the PRP preparation and on the

  4. Platelet function in baboons and humans - A comparative study of whole blood using impedance platelet aggregometry (Multiplate®).

    PubMed

    Ponschab, Martin; van Griensven, Martijn; Heitmeier, Stefan; Laux, Volker; Schlimp, Christoph J; Calatzis, Andreas; Bahrami, Soheyl; Redl, Heinz; Schöchl, Herbert

    2016-11-01

    Platelets play a pivotal role in coagulation, inflammation and wound healing. Suitable animal models that have the potential to mimic human platelet function are limited. The objective of the current study was to compare platelet aggregation response in the whole blood of baboons and humans using impedance aggregometry. Blood was drawn from 24 anesthetised male baboons and 25 healthy volunteers. The platelet aggregation response was determined by impedance aggregometry (Multiplate®). Platelets in the hirudinised whole blood samples were stimulated with four different activators: adenosine diphosphate (ADP), collagen (COL), thrombin receptor activating peptide-6 (TR1AP), and activation of PAR-4 thrombin receptor subtype (TR4AP) at standard concentrations. Higher than standard concentrations were tested in a subgroup of the animals. The cell counts showed no differences between baboons and humans. The platelet aggregation response was significantly lower in baboons compared to humans when stimulated with the platelet agonists ADP (p<0.0001), COL (p=0.021) and TR4AP (p<0.0001). TR1AP did not stimulate platelet aggregation in the baboon blood. Doubling the concentration of ADP and of TR4AP significantly increased the AUC compared to the standard concentration. In contrast, increased COL levels did not further increase the AUC. The current study revealed that testing the platelet function in baboon blood by impedance aggregometry is feasible with ADP, COL and TR4AP, but not with TR1AP. Compared to humans, the aggregation response is lower in baboons. Considering the limitations in accordance to these results, baboons might represent a potential species for further platelet research. Copyright © 2016. Published by Elsevier Ltd.

  5. Platelet-rich concentrate in serum free medium enhances osteogenic differentiation of bone marrow-derived human mesenchymal stromal cells

    PubMed Central

    Ramasamy, Thamil Selvee; Karunanithi, Puvanan; Naveen, Sangeetha Vasudevaraj; Murali, Malliga Raman; Abbas, Azlina A.; Kamarul, Tunku

    2016-01-01

    Previous studies have shown that platelet concentrates used in conjunction with appropriate growth media enhance osteogenic differentiation of human mesenchymal stromal cells (hMSCs). However, their potential in inducing osteogenesis of hMSCs when cultured in serum free medium has not been explored. Furthermore, the resulting osteogenic molecular signatures of the hMSCs have not been compared to standard osteogenic medium. We studied the effect of infrequent supplementation (8-day interval) of 15% non-activated platelet-rich concentrate (PRC) in serum free medium on hMSCs proliferation and differentiation throughout a course of 24 days, and compared the effect with those cultured in a standard osteogenic medium (OM). Cell proliferation was analyzed by alamar blue assay. Gene expression of osteogenic markers (Runx2, Collagen1, Alkaline Phosphatase, Bone morphogenetic protein 2, Osteopontin, Osteocalcin, Osteonectin) were analyzed using Q-PCR. Immunocytochemical staining for osteocalcin, osteopontin and transcription factor Runx2 were done at 8, 16 and 24 days. Biochemical assays for the expression of ALP and osteocalcin were also performed at these time-points. Osteogenic differentiation was further confirmed qualitatively by Alizarin Red S staining that was quantified using cetylpyridinium chloride. Results showed that PRC supplemented in serum free medium enhanced hMSC proliferation, which peaked at day 16. The temporal pattern of gene expression of hMSCs under the influence of PRC was comparable to that of the osteogenic media, but at a greater extent at specific time points. Immunocytochemical staining revealed stronger staining for Runx2 in the PRC-treated group compared to OM, while the staining for Osteocalcin and Osteopontin were comparable in both groups. ALP activity and Osteocalcin/DNA level were higher in the PRC group. Cells in the PRC group had similar level of bone mineralization as those cultured in OM, as reflected by the intensity of Alizarin red

  6. Platelets: handle with care.

    PubMed

    Thomas, S

    2016-10-01

    Platelets are delicate cells that require careful handling between collection, preparation and transfusion. This review addresses practical questions relating to platelet concentration, resting time after collection, total time and number of periods without agitation and temperature. The bags in which platelets are stored are made from gas-permeable plastic to allow sufficient oxygen for the platelets to maintain aerobic respiration. Manufacturers have assigned limits for platelet content and concentration, and these must not be exceeded. There is no strong evidence for or against the resting of platelets post-collection and pre-agitation, but platelets should not be over-wrapped during this period as this compromises gas exchange; a short rest period of up to 1 h may allow the separation of minor aggregates. It is necessary to transport platelet concentrates (e.g. from manufacturing site to hospital), but these periods without gas exchange must be limited to avoid excessive damage to the platelets. Current data support a total of 24 h of transportation per component but with no individual period lasting more than 8 h. Platelets need to be stored at 20-24 °C based on evidence that colder storage leads to irreversible changes on the platelet membrane, resulting in phagocytosis of the platelets following transfusion. Storage at warmer temperatures may lead to an increase in bacterial risk. On the basis of this review, the UK Guidelines for Blood Transfusion Services have been updated to ensure that platelets are handled in the most appropriate way to ensure that efficacious components are provided for patients.

  7. Feasibility of using acoustic velocity meters for estimating highly organic suspended-solids concentrations in streams

    USGS Publications Warehouse

    Patino, Eduardo

    1996-01-01

    A field experiment was conducted at the Levee 4 canal site below control structure G-88 in the Everglades agricultural area in northwestern Broward County, Florida, to study the relation of acoustic attenuation to suspended-solids concentrations. Acoustic velocity meter and temperature data were obtained with concurrent water samples analyzed for suspended-solids concentrations. Two separate acoustic velocity meter frequencies were used, 200 and 500 kilohertz, to determine the sensitivity of acoustic attenuation to frequency for the measured suspended-solids concentration range. Suspended-solids concentrations for water samples collected at the Levee 4 canal site from July 1993 to September 1994 ranged from 22 to 1,058 milligrams per liter, and organic content ranged from about 30 to 93 percent. Regression analyses showed that attenuation data from the acoustic velocity meter (automatic gain control) and temperature data alone do not provide enough information to adequately describe the concentrations of suspended solids. However, if velocity is also included as one of the independent variables in the regression model, a satisfactory correlation can be obtained. Thus, it is feasible to use acoustic velocity meter instrumentation to estimate suspended-solids concentrations in streams, even when suspended solids are primarily composed of organic material. Using the most comprehensive data set available for the study (500 kiloherz data), the best fit regression model produces a standard error of 69.7 milligrams per liter, with actual errors ranging from 2 to 128 milligrams per liter. Both acoustic velocity meter transmission frequencies of 200 and 500 hilohertz produced similar results, suggesting that transducers of either frequency could be used to collect attenuation data at the study site. Results indicate that calibration will be required for each acoustic velocity meter system to the unique suspended-solids regime existing at each site. More robust solutions may

  8. The effects of the oral administration of fish oil concentrate on the release and the metabolism of (/sup 14/C)arachidonic acid and (/sup 14/C)eicosapentaenoic acid by human platelets

    SciTech Connect

    Hirai, A.; Terano, T.; Hamazaki, T.; Sajiki, J.; Kondo, S.; Ozawa, A.; Fujita, T.; Miyamoto, T.; Tamura, Y.; Kumagai, A.

    1982-11-01

    It has been suggested by several investigators that eicosapentaenoic acid (C20:5 omega 3, EPA) might have anti-thrombotic effects. In this experiment, the effect of the oral administration of EPA rich fish oil concentrate on platelet aggregation and the release and the metabolism of (/sup 1 -14/C)arachidonic acid and ((U)-/sup 14/C)eicosapentaenoic acid by human platelets was studied. Eight healthy male subjects ingested 18 capsules of fish oil concentrate (EPA 1.4 g) per day for 4 weeks. Plasma and platelet concentrations of EPA markedly increased, while those of arachidonic acid (C20:4 omega 6, AA) and docosahexaenoic acid (C22:6 omega 3, DHA) did not change. Platelet aggregation induced by collagen and ADP was reduced. Collagen induced (/sup 14/C)thromboxane B2 (TXB2) formation from (/sup 14/C)AA prelabeled platelets decreased. There was no detectable formation of (/sup 14/C)TXB3 from (/sup 14/C)EPA prelabeled platelets, and the conversion of exogenous (/sup 14/C)EPA to (/sup 14/C)TXB3 was lower than that of (/sup 14/C)AA to (/sup 14/C)TXB2. The release of (/sup 14/C)AA from (/sup 14/C)AA prelabeled platelets by collagen was significantly decreased. These observations raise the possibility that the release of arachidonic acid from platelet lipids might be affected by the alteration of EPA content in platelets.

  9. Feasibility of Uranium Concentration Measurements for H Canyon Tank 16.7

    SciTech Connect

    Lascola, R.J.

    2003-01-29

    Savannah River Technology Center (SRTC) evaluated the feasibility of using the H Canyon on-line diode array spectrophotometer to measure uranium concentrations in Tank 16.7. On-line measurements will allow an increase in highly enriched uranium (HEU) production by removing delays associated with off-line measurements. The instrument must be able to measure uranium at concentrations below 1.0 g/L with an uncertainty no greater than 0.3 g/L. SRTC determined that the system has a limit of quantitation of 0.15 g/L. At concentrations of 0.5 and 1.0 g/L, the spectrometer uncertainty is 0.10 g/L. No design changes, such as an increase in flow cell path length, are required to obtain this performance. Expected levels of iron in Tank 16.7 solutions will not interfere with the measurement. The CHEMCHEK method should not be used for confirmatory analysis, as it contributes excessively to the overall uncertainty of the measurement. SRTC expects that the spectrophotometer will meet the measurement requirements for Tank 16.7.

  10. Objective evaluation of the effect of autologous platelet concentrate on post-operative scarring in deep burns.

    PubMed

    Klosová, Hana; Stětinský, Jiří; Bryjová, Iveta; Hledík, Stanislav; Klein, Leo

    2013-09-01

    The healing of grafted areas after surgical treatment of deep burns frequently generates mutilating scars, and rises the risk of subsequent scar hypertrophy. Scar assessment based on clinical evaluation is inherently subjective, which stimulates search for objective means of evaluation. The aim of this study was to objectively evaluate the effect of using autologous platelet concentrate (APC) in combination with split thickness skin grafting (STSG) on scarring processes following surgery of deep burns as compared with application of STSG alone. Selected viscoelastic properties of 38 scars on 23 patients in total were examined using the Cutometer MPA 580 under controlled conditions for long-term outcomes 1, 3, 6 and 12 months after surgery following deep burns. The findings of this study suggest that the STSG+APC combination reduces the time of scar viscoelastic properties recovery as compared with application of STSG alone. This was statistically significant for viscoelastic parameters R2 and Q1. APC has been advocated to enhance scarring after surgery of deep dermal and full thickness burns. We objectively demonstrated that the viscoelastic properties of scars treated with STSG+APC combination return more rapidly to the plateau state than areas treated with STSG only. Copyright © 2013 Elsevier Ltd and ISBI. All rights reserved.

  11. Second-generation platelet concentrate (PRF) as a pulpotomy medicament in a permanent molar with pulpitis: a case report.

    PubMed

    Hiremath, H; Saikalyan, S; Kulkarni, S S; Hiremath, V

    2012-01-01

      To discuss the clinical and radiographic success of a pulpotomy with second-generation platelet concentrate (PRF), in a human mature permanent molar tooth.   A 19-year-old female patient reported to the Department of Conservative Dentistry and Endodontics with established pulpitis in tooth 36. The tooth had a carious pulp exposure, with a history of lingering pain. After isolation, caries removal and pulp exposure, pulpotomy with PRF was performed and a permanent restoration was placed immediately. At the first recall (+1 day), no postoperative pain was reported. At 6, 12, 18 and 22 months recall, the tooth responded positively to pulp sensibility tests, and radiographic examination revealed a normal periodontal ligament space. Positive results of this case imply the need for more studies with larger sample sizes and a longer recall period to justify the use of this novel material for the treatment of pulpitis in human permanent molar teeth. Pulpotomy with PRF could be an alternate treatment to mineral trioxide aggregate or other materials in mature permanent teeth with pulpitis. © 2011 International Endodontic Journal.

  12. Agonist-evoked changes in cytosolic pH and calcium concentration in human platelets: studies in physiological bicarbonate.

    PubMed Central

    Sage, S O; Jobson, T M; Rink, T J

    1990-01-01

    1. Cytosolic pH (pHi) and calcium concentration ([Ca2+]i) have been investigated in the presence and absence of physiological HCO3- in human platelets co-loaded with the fluorescent indicators BCECF and Fura-2. Basal pHi and changes evoked by butyrate, thrombin, platelet activating factor (PAF), ADP and phorbol ester were investigated, as were the effects of removing external Na+. 2. In the presence of physiological HCO3- and CO2, basal pHi was 7.02 +/- 0.04 compared with 7.15 +/- 0.05 in the absence of HCO3-. Estimated cytosolic buffering power was reduced from 35.6 +/- 3.0 to 14.5 +/- 0.4 mM/pH unit by the omission of HCO3-. 3. Thrombin evoked an immediate acidification of 0.03 +/- 0.01 pH units in the presence of HCO3- and 0.07 +/- 0.01 pH units in its absence. The acidifications were followed by a slow alkalinization. The final pHi was 0.10 +/- 0.01 units above basal in the presence of HCO3- and 0.08 +/- 0.02 units above basal in the absence of HCO3-. The initial acidification was significantly greater in the absence of HCO3-. The subsequent increase in pHi was similar in the presence and absence of this ion, but the calculated loss of proton equivalents was greater in the presence of HCO3-. 4. Replacement of extracellular Na+ with N-methyl-D-glucamine resulted in a fall in basal pHi and abolished recovery from thrombin-evoked acidification in both the presence and absence of HCO3-. 5. In the presence of HCO3-, PAF and ADP evoked an intracellular acidification similar to that caused by thrombin. However, with PAF and ADP, the subsequent recovery in pHi was slow and did not rise above basal levels. Phorbol dibutyrate, an activator of protein kinase C, evoked a similar elevation in pHi of 0.04 +/- 0.01 units over 3 min in the presence and absence of HCO3-. 6. Stopped-flow fluorimetric measurements were made of both BCECF and Fura-2 fluorescence in the presence of HCO3-. In the presence and absence of external Ca2+, thrombin-evoked rises in [Ca2+]i peaked before

  13. THE PAPWORTH PLUG - successful use of high dose fibrinogen concentrate and platelet concentrate in potential life-threatening complication after cardiopulmonary bypass surgery in a patient with Type 2M Vicenza von Willebrand Disease.

    PubMed

    Amerikanou, R; MacDonald, S; Lawrence, K; Large, S; Besser, M W

    2012-07-01

    Anecdotally, fibrinogen concentrate (FC) has been used as a "universal" haemostatic agent in complex post-cardiopulmonary bypass (CPB) coagulopathy. We present a case where FC and two pools of platelets prevented life-threatening bleeding in a patient with moderate von Willebrand Disease (vWD) immediately post CPB.

  14. Platelet preservation: agitation and containers.

    PubMed

    van der Meer, Pieter F; de Korte, Dirk

    2011-06-01

    For platelets to maintain their in vitro quality and in vivo effectiveness, they need to be stored at room temperature with gentle agitation in gas-permeable containers. The mode of agitation affects the quality of the platelets, and a gentle method of agitation, either a circular or a flat bed movement, provides the best results. Tumblers or elliptical agitators induce platelet activation and subsequent damage. As long as the platelets remain in suspension, the agitation speed is not important. Agitation of the platelet concentrates ensures that the platelets are continuously oxygenated, that sufficient oxygen can enter the storage container and that excess carbon dioxide can be expelled. During transportation of platelet concentrates, nowadays over long distances where they are held without controlled agitation, platelets may tolerate a certain period without agitation. However, evidence is accumulating that during the time without agitation, local hypoxia surrounding the platelets may induce irreversible harm to the platelets. Over the decades, more gas-permeable plastics have been used to manufacture platelet containers. The use of different plastics and their influence on the platelet quality both in vitro and in vivo is discussed. The improved gas-permeability has allowed the extension of platelet storage from 3 days in the early 1980s, to currently at least 7 days. In the light of new developments, particularly the introduction of pathogen reduction techniques, the use of platelet additive solutions and the availability of improved automated separators, further (renewed) research in this area is warranted.

  15. Platelet additive solution - electrolytes.

    PubMed

    Azuma, Hiroshi; Hirayama, Junichi; Akino, Mitsuaki; Ikeda, Hisami

    2011-06-01

    Recent attention to solutions that replace most or all plasma in platelet concentrates, while maintaining satisfactory platelet function, is motivated by the potential of plasma reduction or depletion to mitigate various transfusion-related adverse events. This report considers the electrolytic composition of previously described platelet additive solutions, in order to draw general conclusions about what is required for platelet function and longevity. The optimal concentrations of Na(+) and Cl(-) are 69-115 mM. The presence of both K(+) and Mg(2+) in platelet suspension at nearly physiological concentrations (3-5mM and 1.5-3mM, respectively) is indispensable for good preservation capacity because both electrolytes are required to prevent platelet activation. In contrast to K(+) and Mg(2+), Ca(2+) may not be important because no free Ca(2+) is available in M-sol, which showed excellent platelet preservation capacity at less than 5% plasma concentration. The importance of bicarbonate (approximately 40 mM) can be recognized when the platelets are suspended in additive solution under less than 5% residual plasma concentration.

  16. Bone marrow aspiration concentrate and platelet rich plasma for osteochondral repair in a porcine osteochondral defect model.

    PubMed

    Betsch, Marcel; Schneppendahl, Johannes; Thuns, Simon; Herten, Monika; Sager, Martin; Jungbluth, Pascal; Hakimi, Mohssen; Wild, Michael

    2013-01-01

    Bone marrow aspiration concentrate (BMAC) may possess a high potency for cartilage and osseous defect healing because it contains stem cells and multiple growth factors. Alternatively, platelet rich plasma (PRP), which contains a cocktail of multiple growth factors released from enriched activated thrombocytes may potentially stimulate the mesenchymal stem cells (MSCs) in bone marrow to proliferate and differentiate. A critical size osteochondral defect (10×6 mm) in both medial femoral condyles was created in 14 Goettinger mini-pigs. All animals were randomized into the following four groups: biphasic scaffold alone (TRUFIT BGS, Smith & Nephew, USA), scaffold with PRP, scaffold with BMAC and scaffold in combination with BMAC and PRP. After 26 weeks all animals were euthanized and histological slides were cut, stained and evaluated using a histological score and immunohistochemistry. The thrombocyte number was significantly increased (p = 0.049) in PRP compared to whole blood. In addition the concentration of the measured growth factors in PRP such as BMP-2, BMP-7, VEGF, TGF-β1 and PDGF were significantly increased when compared to whole blood (p<0.05). In the defects of the therapy groups areas of chondrogenic tissue were present, which stained blue with toluidine blue and positively for collagen type II. Adding BMAC or PRP in a biphasic scaffold led to a significant improvement of the histological score compared to the control group, but the combination of BMAC and PRP did not further enhance the histological score. The clinical application of BMAC or PRP in osteochondral defect healing is attractive because of their autologous origin and cost-effectiveness. Adding either PRP or BMAC to a biphasic scaffold led to a significantly better healing of osteochondral defects compared with the control group. However, the combination of both therapies did not further enhance healing.

  17. Bone Marrow Aspiration Concentrate and Platelet Rich Plasma for Osteochondral Repair in a Porcine Osteochondral Defect Model

    PubMed Central

    Betsch, Marcel; Schneppendahl, Johannes; Thuns, Simon; Herten, Monika; Sager, Martin; Jungbluth, Pascal; Hakimi, Mohssen; Wild, Michael

    2013-01-01

    Background Bone marrow aspiration concentrate (BMAC) may possess a high potency for cartilage and osseous defect healing because it contains stem cells and multiple growth factors. Alternatively, platelet rich plasma (PRP), which contains a cocktail of multiple growth factors released from enriched activated thrombocytes may potentially stimulate the mesenchymal stem cells (MSCs) in bone marrow to proliferate and differentiate. Methods A critical size osteochondral defect (10×6 mm) in both medial femoral condyles was created in 14 Goettinger mini-pigs. All animals were randomized into the following four groups: biphasic scaffold alone (TRUFIT BGS, Smith & Nephew, USA), scaffold with PRP, scaffold with BMAC and scaffold in combination with BMAC and PRP. After 26 weeks all animals were euthanized and histological slides were cut, stained and evaluated using a histological score and immunohistochemistry. Results The thrombocyte number was significantly increased (p = 0.049) in PRP compared to whole blood. In addition the concentration of the measured growth factors in PRP such as BMP-2, BMP-7, VEGF, TGF-β1 and PDGF were significantly increased when compared to whole blood (p<0.05). In the defects of the therapy groups areas of chondrogenic tissue were present, which stained blue with toluidine blue and positively for collagen type II. Adding BMAC or PRP in a biphasic scaffold led to a significant improvement of the histological score compared to the control group, but the combination of BMAC and PRP did not further enhance the histological score. Conclusions The clinical application of BMAC or PRP in osteochondral defect healing is attractive because of their autologous origin and cost-effectiveness. Adding either PRP or BMAC to a biphasic scaffold led to a significantly better healing of osteochondral defects compared with the control group. However, the combination of both therapies did not further enhance healing. PMID:23951201

  18. Screening of platelet concentrates for bacterial contamination: spectrum of bacteria detected, proportion of transfused units, and clinical follow-up.

    PubMed

    Walther-Wenke, Gabriele; Schrezenmeier, Hubert; Deitenbeck, Robert; Geis, Gabriele; Burkhart, Jürgen; Höchsmann, Britta; Sireis, Walid; Schmidt, Michael; Seifried, Erhard; Gebauer, Wolfgang; Liebscher, Ute-Maja; Weinauer, Franz; Müller, Thomas H

    2010-01-01

    Screening of platelet concentrates (PCs) for bacterial contamination with cultivation methods is carried out as a routine procedure in some countries. The aim is to prevent the transfusion of contaminated PCs. The German Evaluation of Regular Monitoring Study Group conducted a prospective multicenter study on 52,243 PCs to investigate the prevalence of bacteria (BacT/ALERT, bioMerieux). This study describes the detected bacterial spectrum, the proportion of PCs with a positive test result that had been transfused, and the results of the clinical follow-up. One hundred thirteen (67%) of 169 potentially or confirmed positive units had already been transfused at the time of the first positive signal. The transfusion of units contaminated by Staphylococcus aureus, Serratia marcescens, and 73% of the units contaminated with Staphylococcus epidermidis, Staphylococcus capitis, or Staphylococcus saccharolyticus was prevented. In contrast, 85% of units with Propionibacterium acnes were transfused. A clonal relationship of the isolates from the pooled PCs and from the associated red blood cell concentrates was found in all investigated cases. The follow-up revealed six febrile reactions to culture-positive PCs not classified as transfusion reaction (TRs) by treating physicians. This demonstrates the importance of hemovigilance. Serious septic reactions due to Klebsiella pneumoniae in two units of one apheresis PC that had tested false-negative were reported; one had a fatal outcome. Culture systems reduce the risk of transfusion of contaminated PCs but cannot guarantee sterility. Physicians must be aware of bacterial contamination of PCs as a potential cause of TRs and must report all adverse events.

  19. Evaluation of the Sensitivity and Specificity of Use of Glucose and pH for Bacterial Screening of Platelet Concentrates Compared to the Bact/Alert.

    PubMed

    Razjou, Farhad; Naghadeh, Hossein Timori; Ferdowsi, Shirin; Dabirmoghadam, Abolfazl

    2017-03-01

    Bacterial contamination of blood components is the major infectious risk in transfusion medicine. Since platelets should be stored at room temperature that makes them an excellent growth medium for bacteria; it is mentioned as a major problem in transfusion medicine. Transfusion risk of a bacterial contaminated platelet concentrate is higher than viral pathogen such as HIV, HBV, HCV and HTLV. The objective of this study was to evaluation of the sensitivity and specificity of use of glucose and pH for bacterial screening of platelet concentrates compared to the Bact/Alert. 1332 platelet concentrates were screened by the Bact/Alert system for aerobic and anaerobic bacterial contamination. Bacterial contamination was also evaluated by using urine reagent strips (Multistix10 SG Bayer) and culture methods. Moreover PH screening with a pH meter (Metrohm 744 Swiss) and glucose was also used for detection of bacterial contamination. The rate of bacterial contamination detected by the Bact/Alert system in platelet concentrates was 25 in 1332 (1.9 %). It contained 15 (1.1 %) for aerobic bacteria and 10 (.8 %) for anaerobic bacteria. 226 of 1332 were considered as containing bacteria by using urine reagent strips. Six of the 226 units were also positive by the Bact/Alert system. Three of those units were culture positive for aerobic bacteria and three for anaerobic. The result of platelet concentrates that underwent pH screening by use of pH meter and a pH portion of urine reagent strips was the same. The sensitivity and specificity of considering glucose alone for detection of bacterial contamination were 12 and 98 % respectively. For pH alone, these were 24 and 83 %. For glucose and/or pH, these were 24 and 83 %; and for combination of glucose and pH, these were 12 and 98 %. Our results showed use of glucose/pH strips would improve the safety of blood products and should be encouraged.

  20. Reactions Induced by Platelet Transfusions

    PubMed Central

    Kiefel, Volker

    2008-01-01

    Summary Platelet transfusions play a central role in therapeutic regimens for patients with hematologic/oncologic diseases who develop severe thrombocytopenia either in the course of their disease or following cytostatic therapy. Like other blood components, platelet transfusions have achieved a high degree of safety as far as transmission of viral diseases is concerned. However, transfusion of platelet concentrates is accompanied by a high frequency of febrile and anaphylactoid reactions. In rare cases, recipients of platelet concentrates are threatened by severe reactions as septic complications due to bacterial contamination of platelet concentrates, transfusion-related acute lung injury and severe anaphylactic episodes. PMID:21512624

  1. Replaced platelet concentrates containing a new additive solution, M-sol: safety and efficacy for pediatric patients.

    PubMed

    Yanagisawa, Ryu; Shimodaira, Shigetaka; Kojima, Shunsuke; Nakasone, Nobuhiko; Ishikawa, Shinsuke; Momose, Kayo; Honda, Takayuki; Yoshikawa, Kentaro; Saito, Shoji; Tanaka, Miyuki; Nakazawa, Yozo; Sakashita, Kazuo; Shiohara, Masaaki; Akino, Mitsuaki; Hirayama, Junichi; Azuma, Hiroshi; Koike, Kenichi

    2013-09-01

    Allergic transfusion reactions (ATRs), particularly those caused by plasma-rich platelet concentrates (P-PCs), are an important concern in transfusion medicine. Replacing P-PCs with PCs containing M-sol (M-sol-R-PCs) is expected to prevent ATRs. However, this has not yet been verified by sufficient clinical evidence. A retrospective cohort study was performed between 2008 and 2011. Pediatric patients with hematologic disorders, solid tumors, primary immunodeficiency disorders, or inherited metabolic disorders were transfused with M-sol-R-PCs between 2010 and 2011; the transfusions of P-PCs administered between 2008 and 2011 were compared in terms of frequency and severity of ATRs, corrected count increment (CCI), and occurrence of bleeding. Data were collected for 6 consecutive months on a per-patient basis. Data obtained during 2008 to 2011 showed that of the 78 patients receiving 515 P-PC transfusions, 14 (17.9%) had 17 ATRs (3.3%); 14 and three ATRs were of Grades 1 and 2, respectively. In 2010 to 2011, 49 patients received 620 transfusions of M-sol-R-PCs, and two patients (4.1%) had Grade 1 ATRs (0.3%). Thus, the frequency of ATRs per bag and per patient differed significantly between the two transfusions. No steroid agents were used for the prevention or treatment of ATRs in the M-sol-R-PC group. The CCI (24 hr) for M-sol-R-PCs did not differ from that for P-PCs. M-sol-R-PCs were found to be effective in preventing ATRs without loss of transfusion efficiency in children; however, its efficacy should be further evaluated in prospective clinical trials. © 2012 American Association of Blood Banks.

  2. Transcrestal Sinus Lift Using Platelet Concentrates in Association to Short Implant Placement: A Retrospective Study of Augmented Bone Height Remodeling.

    PubMed

    Anitua, Eduardo; Flores, Javier; Alkhraisat, Mohammad Hamdan

    2016-10-01

    No evidence has been reported yet about dimensional changes in maxillary sinuses grafted with autologous platelet concentrate. This study aimed to analyze the augmented height alterations around short implants (length ≤ 8.5 mm) placed after transcrestal sinus lift surgery in association with plasma rich in growth factors. A retrospective design was used. Patients with atrophic posterior maxilla were treated by inserting short implants in combination with transalveolar maxillary sinus floor augmentation. Radiographic bone level alterations over time were assessed on panoramic radiographs. Transalveolar sinus augmentation was performed to place 41 implants in 26 patients with a residual bone height of 4.7 ± 1.3 mm. The alveolar bone height was increased by 3.7 ± 1.7 mm and 4.2 ± 2.0 mm after 12 ± 3 months and 35 ± 11 months of surgery, respectively, whereas the apical augmented height (beyond the implant) apex was stable around 34 implants during the follow-up. It was increasing around seven implants that were not covered by the endo-sinus augmented height at the 1-year follow-up. Transalveolar sinus floor augmentation in association with plasma rich in growth factors and short implants resulted in a stable augmented height gain after 3 years follow-up. Atrophic posterior maxilla could be treated by transalveolar sinus lift in association with plasma rich in growth factors and the placement of short implants. © 2015 Wiley Periodicals, Inc.

  3. Chronic wounds treated with a physiologically relevant concentration of platelet-rich plasma gel: a prospective case series.

    PubMed

    Frykberg, Robert G; Driver, Vickie R; Carman, Donna; Lucero, Brenda; Borris-Hale, Cathy; Fylling, Carelyn P; Rappl, Laurie M; Clausen, Peter A

    2010-06-01

    Chronic wounds are characterized by a long inflammatory phase that hinders regenerative wound healing. The purpose of this prospective case series was to evaluate how a physiologically relevant concentration of an autologous platelet-rich plasma (PRP) gel affects initial wound healing trajectories of chronic, nonhealing wounds of various etiologies and in different care settings. Using convenience sampling methods, 49 patients (average age: 60.6 years, SD 14.7) with 65 nonhealing wounds (mean duration 47.8 weeks, range 3 to 260) at eight long-term acute care (LTAC) hospitals and three outpatient foot or wound clinics who were prescribed PRP gel for their nonhealing wound were enrolled. The majority of patients had low albumin, hematocrit, and/or hemoglobin levels. After wound assessments and measurements were obtained and the gel prepared, a skin barrier was applied to the periwound skin and the gel applied and protected with cover dressings. The most common wounds were pressure ulcers (n = 21), venous ulcers (n = 16) and diabetic foot ulcers (n = 14). Mean wound area and volume were 19 cm2 (SD 29.4) and 36.2 cm3 (SD 77.7), respectively. Following a mean of 2.8 (SD 2.4) weeks with 3.2 (SD 2.2) applications, reductions in wound volume (mean 51%, SD 43.1), area (39.5%, SD 41.2), undermining (77.8%, SD 28.9), and sinus tract/tunneling (45.8%, SD 40.2) were observed. For all wound etiologies, 97% of wounds improved. The results of this study suggest the application of this PRP gel can reverse nonhealing trends in chronic wounds.

  4. The Pan Genera Detection immunoassay: a novel point-of-issue method for detection of bacterial contamination in platelet concentrates.

    PubMed

    Vollmer, Tanja; Hinse, Dennis; Kleesiek, Knut; Dreier, Jens

    2010-10-01

    Bacterial contamination of platelet concentrates (PCs) still represents an ongoing risk in transfusion-transmitted sepsis. Recently the Pan Genera Detection (PGD) system was developed and FDA licensed for screening of bacterial contamination of PCs directly prior to transfusion. The test principle is based on the immunological detection of lipopolysaccharide (for Gram-negative bacteria) or lipoteichoic acid (for Gram-positive bacteria). In the present study we analyzed the applicability of this method with regard to detection limit, practicability, implementation, and performance. PCs were spiked with Staphylococcus aureus, Bacillus subtilis, and five different Klebsiella pneumoniae strains, as well as eight different Escherichia coli strains. The presence of bacteria was assessed by the PGD immunoassay, and bacteria were enumerated by plating cultures. Application of the PGD immunoassay showed that it is a rapid test with a short hands-on time for sample processing and no demand for special technical equipment and instrument operation. The lower detection limits of the assay for Gram-positive bacteria showed a good agreement with the manufacturer's specifications (8.2 × 10(3) to 5.5 × 10(4) CFU/ml). For some strains of K. pneumoniae and E. coli, the PGD test showed analytical sensitivities (>10(6) CFU/ml) that were divergent from the designated values (K. pneumoniae, 2.0 × 10(4) CFU/ml; E. coli, 2.8 × 10(4) CFU/ml). Result interpretation is sometimes difficult due to very faint bands. In conclusion, our study demonstrates that the PGD immunoassay is an easy-to-perform bedside test for the detection of bacterial contamination in PCs. However, to date there are some shortcomings in the interpretation of results and in the detection limits for some strains of Gram-negative bacteria.

  5. Routine bacterial screening of platelet concentrates by flow cytometry and its impact on product safety and supply.

    PubMed

    Müller, B; Walther-Wenke, G; Kalus, M; Alt, T; Bux, J; Zeiler, T; Schottstedt, V

    2015-04-01

    Bacterial contamination represents the major infectious hazard associated with transfusion of platelet concentrates (PCs). As bacterial screening of PCs is not mandatory in Germany, the BactiFlow flow cytometry test has been introduced as a rapid detection method to increase product safety. During a period of 25 months, a total of 34 631 PCs (26 411 pooled and 8220 apheresis-derived PCs) were tested at the end of day 3 of their shelf life using the BactiFlow system. PCs initially reactive in BactiFlow testing and expired PCs not reactive in BactiFlow on day 3 were also investigated by the BacT/ALERT system and by microbiological cultivation in order to identify the contaminating bacterial species and to confirm reactive BactiFlow results. Two hundred and twenty-eight PCs (0.7%) had an initially reactive result, 24 of them remained reactive in a second test run. Out of these reproducible reactive BactiFlow results, 12 could not be verified by parallel BacT/ALERT culturing, resulting in a confirmed false-positive rate of 0.03%. The bacterial species were identified as S. aureus, S. epidermidis, S. dysgalactiae ssp. equisimilis and B. cereus. In 10 out of 9017 expired PCs (0.11%), a confirmed-positive result was obtained in the BacT/ALERT system which had a negative result in the BactiFlow system. Testing of PCs by BactiFlow was successfully implemented in our blood donation service and proved sufficient as a rapid and reliable screening method. False reactive results are in an acceptable range since the transfusion of 12 bacterially contaminated PCs was prevented. © 2014 International Society of Blood Transfusion.

  6. Addition of platelet concentrate to Dermo-Epidermal Skin Graft in deep burn trauma reduces scarring and need for revision surgeries

    PubMed Central

    Prochazka, Vaclav; Klosova, Hana; Stetinsky, Jiri; Gumulec, Jaromir; Vitkova, Katerina; Salounova, Dana; Dvorackova, Jana; Bielnikova, Hana; Klement, Petr; Levakova, Veronika; Ocelka, Tomas; Pavliska, Lubomir; Kovanic, Pavel; Klement, Giannoula Lakka

    2017-01-01

    Backround Deep skin burn injuries, especially those on the face, hands, feet, genitalia and perineum represent significant therapeutic challenges. Autologous dermo-epidermal skin grafts (DESG) have become standard of care for treating deep burns. Additionally, human autologous thrombin activated autologous platelet concentrate (APC) has gained acceptance in the setting of wounds. While each of these interventions has been independently shown to accelerate healing, the combination of the two has never been evaluated. We hypothesized that the addition of platelets (source of growth factors and inhibitors necessary for tissue repair) to the DESG (source of progenitor cells and of tissue proteases necessary for spatial and temporal control of growth regulators released from platelets) would create the optimal environment for the reciprocal interaction of cells within the healing tissues. Methods We used clinical examination (digital photography), standardised scales for evaluating pain and scarring, in combination with blood perfusion (laser Doppler imaging), as well as molecular and laboratory analyses. Results We show for the first time that the combination of APC and DESG leads to earlier relief of pain, and decreased use of analgesics, antipruritics and orthotic devices. Most importantly, this treatment is associated with earlier discharges from hospital and significant cost savings. Conclusions Our findings indicate that DESG engraftment is facilitated by the local addition of platelets and by systemic thrombocytosis. This local interaction leads to the physiological revascularization at 1–3 months. We observed significant elevation of circulating platelets in early stages of engraftment (1–7 days), which normalized over the subsequent 7 and 90 days. PMID:24108222

  7. Platelet interactions with Candida albicans.

    PubMed Central

    Skerl, K G; Calderone, R A; Sreevalsan, T

    1981-01-01

    The interaction of human platelets and Candida albicans was studied. Platelet-rich plasma was obtained from freshly drawn blood or outdated platelet concentrates. From the platelet-rich plasma, a platelet extract was derived which stimulated germ tube formation by C. albicans when incubated with yeast cells at 37 degrees C. The active component(s) was heat stable, trypsin sensitive, and ribonuclease and deoxyribonuclease insensitive, and possessed cationic properties since it readily attached to carboxymethyl-Sephadex. The active component(s) seemed to bind to heparin also, since germ tube-promoting activity was eluted from a heparin-cyanogen bromide-activated Sepharose 4B column. In addition, platelet-derived growth factor (Collaborative Research, Inc.) stimulated germination when incubated with low amounts (0.4% final concentration) of bovine calf serum. The aggregation of platelets, prepared as platelet-rich plasma by C. albicans cell wall or alkali-extracted cell wall fractions, was also studied. Aggregation of platelets was observed when cell wall or cell wall fractions were incubated with platelet-poor plasma at 37 degrees C for 20 min and then added to platelet-rich plasma. The component of platelet-poor plasma which promoted aggregation of platelets by C. albicans cell wall or alkali-extracted fractions was inactivated at 56 degrees C (30 min) and by cobra venom factor, indicating a role for the alternate complement pathway in the aggregation response. PMID:7037646

  8. Surface modification of CoCr alloy using varying concentrations of phosphoric and phosphonoacetic acids: albumin and fibrinogen adsorption, platelet adhesion, activation, and aggregation studies.

    PubMed

    Thiruppathi, Eagappanath; Larson, Mark K; Mani, Gopinath

    2015-01-01

    CoCr alloy is commonly used in various cardiovascular medical devices for its excellent physical and mechanical properties. However, the formation of blood clots on the alloy surfaces is a serious concern. This research is focused on the surface modification of CoCr alloy using varying concentrations (1, 25, 50, 75, and 100 mM) of phosphoric acid (PA) and phosphonoacetic acid (PAA) to generate various surfaces with different wettability, chemistry, and roughness. Then, the adsorption of blood plasma proteins such as albumin and fibrinogen and the adhesion, activation, and aggregation of platelets with the various surfaces generated were investigated. Contact angle analysis showed PA and PAA coatings on CoCr provided a gradient of hydrophilic surfaces. FTIR showed PA and PAA were covalently bound to CoCr surface and formed different bonding configurations depending on the concentrations of coating solutions used. AFM showed the formation of homogeneous PA and PAA coatings on CoCr. The single and dual protein adsorption studies showed that the amount of albumin and fibrinogen adsorbed on the alloy surfaces strongly depend on the type of PA and PAA coatings prepared by different concentrations of coating solutions. All PA coated CoCr showed reduced platelet adhesion and activation when compared to control CoCr. Also, 75 and 100 mM PA-CoCr showed reduced platelet aggregation. For PAA coated CoCr, no significant difference in platelet adhesion and activation was observed between PAA coated CoCr and control CoCr. Thus, this study demonstrated that CoCr can be surface modified using PA for potentially reducing the formation of blood clots and improving the blood compatibility of the alloy.

  9. Feasibility of the Simultaneous Determination of Monomer Concentrations and Particle Size in Emulsion Polymerization Using in Situ Raman Spectroscopy

    PubMed Central

    2015-01-01

    An immersion Raman probe was used in emulsion copolymerization reactions to measure monomer concentrations and particle sizes. Quantitative determination of monomer concentrations is feasible in two-monomer copolymerizations, but only the overall conversion could be measured by Raman spectroscopy in a four-monomer copolymerization. The feasibility of measuring monomer conversion and particle size was established using partial least-squares (PLS) calibration models. A simplified theoretical framework for the measurement of particle sizes based on photon scattering is presented, based on the elastic-sphere-vibration and surface-tension models. PMID:26900256

  10. Persistent aggregates in apheresis platelet concentrates are commonly collected from donors with a history of aggregate donation.

    PubMed

    Feys, H B; Pottel, H; Coene, J; Vandewalle, G; Vandekerckhove, P; Compernolle, V

    2016-11-01

    Platelet apheresis sometimes causes persistent aggregates (PA). This study (n = 211) shows that changing the apheresis settings to reach fixed product volumes instead of yields does not influence PA incidence, even though PA products on average contain more platelets than controls. Furthermore, logistic regression was used to model if PA can be predicted on the basis of certain predonation parameters. PA donation history was the only parameter retained, proving a strong determinant of predictability [AUC = 0.735 (SE = 0.022)]. Consequently, donations from a donor with previous PA history are 7.8 times more likely to contain PA than from a donor without preceding history.

  11. Short term storage stability at room temperature of two different platelet-rich plasma preparations from equine donors and potential impact on growth factor concentrations.

    PubMed

    Hauschild, Gregor; Geburek, Florian; Gosheger, Georg; Eveslage, Maria; Serrano, Daniela; Streitbürger, Arne; Johannlükens, Sara; Menzel, Dirk; Mischke, Reinhard

    2017-01-05

    The increasing interest in platelet-rich plasma (PRP) based therapies is as yet accompanied by inconsistent information regarding nearly all aspects of handling and application. Among these storage stability of processed platelet-rich products may be the basis for a more flexible application mode. The objective of this study was (1) to estimate the storage stability of growth factors platelet derived growth factor BB (PDGF-BB) and transforming growth factor ß1 (TGF-ß1) in both, a single-step softspin centrifugation-based pure-PRP (P-PRP, ACP®), and a gravity filtration system-based leukocyte-rich-PRP (L-PRP, E-PET), over a six hours time span after preparation at room temperature and (2) to identify possible factors influencing these growth factor concentrations in an equine model. Growth factor concentrations remained stable over the entire investigation period in L-PRP as well as P-PRP preparations revealing a mean of 3569 pg/ml PDGF-BB for E-PET and means of 1276 pg/ml PDGF-BB and 5086 pg/ml TGF-ß1 for ACP®. Pearson correlations yielded no significant impact of whole blood platelet (PLT), white blood cell (WBC) and red blood cell (RBC) counts on resulting cytokine values. In case of ACP® no significant dependencies between PLT, WBC and RBC counts of the processed platelet-rich product and resulting cytokine content occurred with exception of TGF-ß1 concentrations showing a strong correlation with the WBC content. PDGF-BB content of E-PET preparations showed a strong positive correlation with PLT and a strong negative with WBC of these preparations but not with RBC. L-PRP ad modum E-PET and P-PRP ad modum ACP® are applicable over at least a six hours time span at room temperature without loss of growth factor content. Based on the results of this study factors influencing the resulting growth factor concentrations still remain questionable. Additional studies implicating a further standardization of preparation protocols are necessary to identify

  12. Accumulation of CD62P during storage of apheresis platelet concentrates and the role of CD62P in transfusion-related acute lung injury.

    PubMed

    Tong, Shan; Wang, Haibao; Zhang, Ting; Chen, Linfeng; Liu, Bowei

    2015-11-01

    Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion-associated morbidity and mortality. Activated platelets have important roles in TRALI and CD62P was identified to be an important indicator of platelet activation. However, the precise roles of CD62P in TRALI have remained elusive. The present study assessed CD62P accumulation during storage of apheresis platelet concentrates (A‑Plts) and established a mouse model of TRALI to further investigate the roles of CD62P in TRALI. The results showed that the CD62P concentration in A‑Plts was increased with the storage time. Mice were treated with monoclonal major histocompatibility complex (MHC)‑1 antibody to induce TRALI. The murine model of TRALI was successfully established as evidenced by pulmonary oedema, accompanied by decreased clearance of bronchoalveolar lavage fluid (BALF), increased pulmonary and systemic inflammation, elevated lung myeloperoxidase (MPO) activity as well as increased pulmonary and systemic coagulation in the TRALI group compared with those in the control group. To further determine the role of CD62P in TRALI, mice were treated with anti‑CD62P antibody to knockdown CD62P in vivo. It was found that pulmonary oedema, BALF clearance, pulmonary and systemic inflammation, MPO activity as well as pulmonary and systemic coagulation were decreased in the TRALI + anti‑CD62P antibody group compared with those in the TRALI + isotype antibody group. The present study supported the notion that CD62P is involved in mediating TRALI and may provide an important molecular basis for enhancing the clinical safety and effectiveness of platelet transfusion.

  13. Picomolar platelet-activating factor mobilizes Ca to change platelet shape without activating phospholipase C or protein kinase C; simultaneous fluorometric measurement of intracellular free Ca concentration and aggregation.

    PubMed

    James-Kracke, M R; Sexe, R B; Shukla, S D

    1994-11-01

    The purpose of this study was to investigate signal transduction mechanisms activated by low and high concentrations of platelet-activating factor (PAF) in rabbit platelets and to contrast the responses to those induced by thrombin. We measured changes in intracellular free calcium ([Ca++]i) with fura2, while monitoring light scatter simultaneously as a measure of shape change and aggregation in a dual-excitation dual-emission spectrofluorometer. An abrupt 20% fall in light scatter, coincident with the peak of the [Ca++]i, indicated shape change in Ca-containing or Ca-free medium and was blocked by BAPTA loading and 10 microM cytochalasin B. A secondary decline in light scatter, indicating aggregation, occurred only in Ca-containing medium and only under conditions favoring protein kinase C (PKC) activation. PAF at 10(-12) M did not increase 1,4,5-inositol triphosphate content, which suggested PKC would not be activated. However, PAF at 10(-12) rapidly increased [Ca++]i to 900 nM in 7 sec seemingly by Ca influx through receptor-operated channels inducing shape change. PAF at 10(-9) and 10(-8) M increased [Ca++]i to 2 microM in 12 sec and induced both shape change and aggregation. However, in platelets pretreated with 100 nM staurosporine to inhibit protein kinases, 10(-9) M PAF did not cause aggregation even though [Ca++]i still rose to 2 microM, which indicated that PKC plays a role in aggregation but not in Ca++ mobilization.(ABSTRACT TRUNCATED AT 250 WORDS)

  14. Effect of Leukocyte Concentration on the Efficacy of Platelet-Rich Plasma in the Treatment of Knee Osteoarthritis.

    PubMed

    Riboh, Jonathan C; Saltzman, Bryan M; Yanke, Adam B; Fortier, Lisa; Cole, Brian J

    2016-03-01

    Leukocyte-poor platelet-rich plasma (LP-PRP) is hypothesized to be more suitable for intra-articular injection than leukocyte-rich PRP (LR-PRP) in the treatment of knee osteoarthritis. To compare clinical outcomes and rates of adverse reactions between LP-PRP and LR-PRP for this application. Meta-analysis. The MEDLINE, EMBASE, and Cochrane databases were reviewed. The primary outcome was the incidence of local adverse reactions. Secondary outcomes were the changes in International Knee Documentation Committee (IKDC) subjective score and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score between baseline and final follow-up measurements. A Bayesian network meta-analysis was performed, with a post hoc meta-regression to correct for baseline differences in WOMAC scores. Treatment rankings were based on surface under the cumulative ranking (SUCRA) probabilities. Included in the analysis were 6 randomized controlled trials (evidence level 1) and 3 prospective comparative studies (evidence level 2) with a total of 1055 patients. Injection of LP-PRP resulted in significantly better WOMAC scores than did injection of hyaluronic acid (mean difference, -21.14; 95% CI, -39.63 to -2.65) or placebo (mean difference, -17.84; 95% CI, -34.95 to -0.73). No such difference was observed with LR-PRP (mean difference, -14.28; 95% CI, -44.80 to 16.25). All treatment groups resulted in equivalent IKDC subjective scores. The SUCRA analysis showed that LP-PRP was the highest ranked treatment for both measures of clinical efficacy (WOMAC and IKDC). Finally, PRP injections resulted in a higher incidence of adverse reactions than hyaluronic acid (odds ratio, 5.63; 95% CI, 1.38-22.90), but there was no difference between LR-PRP and LP-PRP (odds ratio, 0.78; 95% CI, 0.05-11.93). These reactions were nearly always local swelling and pain, with a single study reporting medical side effects including syncope, dizziness, headache, gastritis, and tachycardia (17/1055 total

  15. Inter-laboratory comparison of different rapid methods for the detection of bacterial contamination in platelet concentrates.

    PubMed

    Vollmer, T; Hinse, D; Schottstedt, V; Bux, J; Tapernon, K; Sibrowski, W; Knabbe, C; Dreier, J

    2012-07-01

      Bacterial contamination of platelet concentrates still represents a major risk in transfusion medicine, and a variety of screening methods have been available to improve the safety of PCs. In the present study, the analytical quality of three different rapid screening methods (BactiFlow flow cytometry, Pan Genera Detection Assay, 23S rRNA RT-PCR) was evaluated in an inter-laboratory comparison in three different German blood services.   Samples were inoculated with different bacteria [Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli (two strains), Klebsiella pneumoniae (two strains), Enterobacter aerogenes (one strain), Serratia marcescens (one strain)] at different counts (4·5 × 10(3) -4·5 × 10(8)  CFU/ml) alternating with negative samples in one transfusion facility. Samples were blinded with a random order for each screening method, shipped to partners and analysed immediately after receipt with different rapid screening methods.   The inter-laboratory comparison revealed that the BactiFlow assay and 23S rRNA RT-PCR-screening detected all samples correctly (positive: 12/12, negative: 8/8). The Pan Genera Detection Assay test detected only four of the positive samples. Four of the non-detected positive samples were below the assay's detection limit. Another four inoculated samples with comparatively high bacteria counts were detected false negative (E. coli (two strains): 9·87 × 10(5) and 2·10 × 10(7)  CFU/ml, respectively, K. pneumoniae: 4·79 × 10(6)  CFU/ml, S. aureus: 6·03 × 10(5)  CFU/ml). All rapid screening methods revealed no false-positive results.   Both BactiFlow and 23S rRNA RT-PCR demonstrated a high sensitivity to detecting bacterial contamination in PCs. The Pan Genera Detection Assay had some shortcomings regarding sensitivity, especially for the detection of Gram-negative strains. © 2011 The Author(s). Vox Sanguinis © 2011 International Society of Blood Transfusion.

  16. Effect of platelet-rich plasma (PRP) concentration on the viability and proliferation of alveolar bone cells: an in vitro study.

    PubMed

    Choi, B-H; Zhu, S-J; Kim, B-Y; Huh, J-Y; Lee, S-H; Jung, J-H

    2005-06-01

    Previous studies have shown that a combination of platelet-rich plasma (PRP) and autogenous bone graft can increase the rate of osteogenesis and enhance bone formation qualitatively. However, contradictory results were reported in a recent animal study. In order to clarify this inconsistency, this study examined the influence of the PRP concentrations on the viability and proliferation of alveolar bone cells in vitro. Bone cells obtained from the alveolar bone chips were exposed to various PRP concentrations. After a culture period of 7 days, cellular viability and proliferation were evaluated by counting the number of cells and a MTT assay. The results showed that the viability and proliferation of alveolar bone cells were suppressed by high PRP concentrations, but were stimulated by low PRP concentrations (1-5%). These in vitro results support the view that variations in the PRP concentrations might influence the bone formation within the PRP-treated bone grafts.

  17. Inhibitory Effects of Cytosolic Ca2+ Concentration by Ginsenoside Ro Are Dependent on Phosphorylation of IP3RI and Dephosphorylation of ERK in Human Platelets

    PubMed Central

    Kwon, Hyuk-Woo; Shin, Jung-Hae; Lee, Dong-Ha; Park, Hwa-Jin

    2015-01-01

    Intracellular Ca2+ ([Ca2+]i) is platelet aggregation-inducing molecule and is involved in activation of aggregation associated molecules. This study was carried out to understand the Ca2+-antagonistic effect of ginsenoside Ro (G-Ro), an oleanane-type saponin in Panax ginseng. G-Ro, without affecting leakage of lactate dehydrogenase, dose-dependently inhibited thrombin-induced platelet aggregation, and the half maximal inhibitory concentration was approximately 155 μM. G-Ro inhibited strongly thrombin-elevated [Ca2+]i, which was strongly increased by A-kinase inhibitor Rp-8-Br-cAMPS compared to G-kinase inhibitor Rp-8-Br-cGMPS. G-Ro increased the level of cAMP and subsequently elevated the phosphorylation of inositol 1, 4, 5-triphosphate receptor I (IP3RI) (Ser1756) to inhibit [Ca2+]i mobilization in thrombin-induced platelet aggregation. Phosphorylation of IP3RI (Ser1756) by G-Ro was decreased by PKA inhibitor Rp-8-Br-cAMPS. In addition, G-Ro inhibited thrombin-induced phosphorylation of ERK 2 (42 kDa), indicating inhibition of Ca2+ influx across plasma membrane. We demonstrate that G-Ro upregulates cAMP-dependent IP3RI (Ser1756) phosphorylation and downregulates phosphorylation of ERK 2 (42 kDa) to decrease thrombin-elevated [Ca2+]i, which contributes to inhibition of ATP and serotonin release, and p-selectin expression. These results indicate that G-Ro in Panax ginseng is a beneficial novel Ca2+-antagonistic compound and may prevent platelet aggregation-mediated thrombotic disease. PMID:26355658

  18. Taurine and platelet aggregation

    SciTech Connect

    Nauss-Karol, C.; VanderWende, C.; Gaut, Z.N.

    1986-03-01

    Taurine is a putative neurotransmitter or neuromodulator. The endogenous taurine concentration in human platelets, determined by amino acid analysis, is 15 ..mu..M/g. In spite of this high level, taurine is actively accumulated. Uptake is saturable, Na/sup +/ and temperature dependent, and suppressed by metabolic inhibitors, structural analogues, and several classes of centrally active substances. High, medium and low affinity transport processes have been characterized, and the platelet may represent a model system for taurine transport in the CNS. When platelets were incubated with /sup 14/C-taurine for 30 minutes, then resuspended in fresh medium and reincubated for one hour, essentially all of the taurine was retained within the cells. Taurine, at concentrations ranging from 10-1000 ..mu..M, had no effect on platelet aggregation induced by ADP or epinephrine. However, taurine may have a role in platelet aggregation since 35-39% of the taurine taken up by human platelets appears to be secreted during the release reaction induced by low concentrations of either epinephrine or ADP, respectively. This release phenomenon would imply that part of the taurine taken up is stored directly in the dense bodies of the platelet.

  19. Platelet interaction with bacteria. V. Ultrastructure of congenital afibrinogenemic platelets.

    PubMed Central

    Clawson, C. C.; White, J. G.

    1980-01-01

    Platelets from a patient with congenital afibrinogenemia (CA) were tested in their native plasma for reactivity in vitro to Staphylococcus aureus 502A. Previous studies of the interactions between normal human platelets and this organism have shown rapid irreversible aggregation responses in which the bacteria were regularly trapped between aggregating platelets. Engulfment of microbes by single normal platelets in a process akin to phagocytosis was a very rare occurrence. In contrast, CA platelets showed a delayed aggregation response to contact with this microorganism. The CA platelets were also much more sensitive to the concentration of bacteria than were normal platelets. Electron microscopy showed that individual CA platelets often engulfed the stimulatory organism rather than participating in aggregation. Postfixation staining with a colloidal tracer, lanthanum nitrate, indicated that the bacteria were sequestered in the open canalicular system of the CA platelets in a manner analogous to that previously observed with latex particles. Restoration of normal levels of human fibrinogen to the CA platelet-rich plasma corrected the delay in aggregation but did not eliminate the frequent engulfment of bacteria by the CA platelets. These findings indicate that fibrinogen is an important, although not essential, cofactor in the response of human platelets to contact with this common bacterial pathogen. Images p[209]-a Figures 6 and 7 Figures 8 and 9 Figures 10 and 11 Figures 12 and 13 Figure 1 Figure 2 Figure 3 PMID:7350814

  20. Platelet Donation

    MedlinePlus

    ... donating platelets, can I still donate blood? What blood types should donate platelets? Can I donate plasma at ... Community Learn About Blood Blood Facts and Statistics Blood Types Blood Components What Happens to Donated Blood Blood ...

  1. Autologous platelet-labeling in thrombocytopenia

    SciTech Connect

    Sinzinger, H.; Virgolini, I.; Vinazzer, H. )

    1990-11-01

    Field studies performed with peripheral platelets obtained from 6 male volunteers aged 23 to 29 years revealed an extraordinary dependence of labeling efficiency on incubation time and platelet concentration after {sup 111}In-oxine platelet labeling. Since the monitoring of in vivo-platelet function in patients with thrombocytopenia may cause problems due to insufficient labeling results and homologous platelets may show a different in vivo behaviour to autologous ones, we have searched for the minimal amount of platelets necessary to allow appropriate labeling and imaging in patients with thrombocytopenia. In 15 patients with untreated thrombocytopenia aged 14 to 79 years demonstrating a mean peripheral platelet count of 2.509 +/- 1.45 x 10(4) cells/microliters autologous {sup 111}In-oxine platelet labeling was performed. The results indicate that approximately 1 x 10(8) (concentrated) platelets/ml are necessary to obtain an adequate labeling efficiency and recovery. This platelet concentration can be easily achieved by drawing one more Monovette of whole blood per each 5 x 10(4) platelets/microliter peripheral platelet count less than 2 x 10(5)/microliter. It is concluded, that calculation of the required number of platelets in advance, variation of the blood volume drawn and the volume of incubation buffer allow informative, qualitative and quantitative results using autologous platelets. The method presented effectively circumvents the requirement of homologous platelets for radiolabeling in thrombocytopenia.

  2. Autologous Bioscaffolds based on Different Concentrations of Platelet Rich Plasma and Synovial Fluid as a Vehicle for Mesencymal Stem Cells.

    PubMed

    Garate, Ane; Sánchez, Pello; Delgado, Diego; Bilbao, Ane Miren; Muiños-López, Emma; Granero-Moltó, Froilán; Orive, Gorka; Prosper, Felipe; Pedraz, José Luis; Sánchez, Mikel

    2017-09-27

    In the field of tissue engineering, diverse types of bioscaffolds are being developed currently for osteochondral defect applications. In this work, a novel scaffold based on Platelet Rich Plasma and hyaluronic acid with Mesenchymal stem cells (MSCs) has been evaluated to observe its effect on immobilized cells. The bio-scaffolds were prepared by mixing different volumes of synovial fluid (SF) with Platelet Rich Plasma (PRP) from patients obtaining 3 formulations at PRP-SF ratios of 3:1, 1:1 and 1:3 (vol/vol). The live/dead staining revealed that although the cell number of each type of bioscaffold was different, this constructs provide cells with a suitable environment for their viability and proliferation. Moreover, immobilized MSCs showed their ability to secrete fibrinolytic enzymes, which vary depending on the fibrin amount of the scaffold. Immunohistochemical analysis revealed the positive staining for collagen type II in all cases, proving the biologic action of synovial fluid derived MSCs together with the suitable characteristics of the bioscaffold for chondrogenic differentiation. Considering all these aspects, this study demonstrates that these cells-based constructs respresent an attractive method for cell immobilization, achieving completely autologous and biocompatible scaffolds. This article is protected by copyright. All rights reserved. © 2017 Wiley Periodicals, Inc.

  3. Positive effect of an autologous platelet concentrate in lateral epicondylitis in a double-blind randomized controlled trial: platelet-rich plasma versus corticosteroid injection with a 1-year follow-up.

    PubMed

    Peerbooms, Joost C; Sluimer, Jordi; Bruijn, Daniël J; Gosens, Taco

    2010-02-01

    Platelet-rich plasma (PRP) has shown to be a general stimulation for repair. Purpose To determine the effectiveness of PRP compared with corticosteroid injections in patients with chronic lateral epicondylitis. Randomized controlled trial; Level of evidence, 1. The trial was conducted in 2 teaching hospitals in the Netherlands. One hundred patients with chronic lateral epicondylitis were randomly assigned in the PRP group (n = 51) or the corticosteroid group (n = 49). A central computer system carried out randomization and allocation to the trial group. Patients were randomized to receive either a corticosteroid injection or an autologous platelet concentrate injection through a peppering technique. The primary analysis included visual analog scores and DASH Outcome Measure scores (DASH: Disabilities of the Arm, Shoulder, and Hand). Successful treatment was defined as more than a 25% reduction in visual analog score or DASH score without a reintervention after 1 year. The results showed that, according to the visual analog scores, 24 of the 49 patients (49%) in the corticosteroid group and 37 of the 51 patients (73%) in the PRP group were successful, which was significantly different (P <.001). Furthermore, according to the DASH scores, 25 of the 49 patients (51%) in the corticosteroid group and 37 of the 51 patients (73%) in the PRP group were successful, which was also significantly different (P = .005). The corticosteroid group was better initially and then declined, whereas the PRP group progressively improved. Treatment of patients with chronic lateral epicondylitis with PRP reduces pain and significantly increases function, exceeding the effect of corticosteroid injection. Future decisions for application of the PRP for lateral epicondylitis should be confirmed by further follow-up from this trial and should take into account possible costs and harms as well as benefits.

  4. Effects of pH and concentration of sodium citrate anticoagulant on platelet aggregation measured by light transmission aggregometry induced by adenosine diphosphate.

    PubMed

    Germanovich, Ksenia; Femia, Eti Alessandra; Cheng, Chun Yan; Dovlatova, Natalia; Cattaneo, Marco

    2017-06-23

    The 2013 ISTH-SSC guidelines for the standardization of light transmission aggregometry (LTA) were largely based on expert consensus, as studies directly comparing LTA methodologies were lacking. We experimentally tested the cogency of ISTH-SSC recommendations pertaining to use of anticoagulant, in particular whether: (1) buffered citrate (BC) is preferable to unbuffered citrate (C); (2) the two recommended concentrations of sodium citrate (109 and 129 mM) are equivalent in terms of platelet aggregation (PA). Blood from 16 healthy volunteers was collected into BC and C (109 and 129 mM). PA was measured by LTA in platelet-rich plasma (PRP) stimulated by adenosine diphosphate (ADP) (2 μM) immediately after PRP preparation and up to 4 hr after blood collection; pH and platelet counts in PRP were measured in parallel. pH in PRP increased with time up to about 8 for all anticoagulants, although it was lower in BC than in C at all times. In BC, PA was lower at 45 min, but equivalent at all other times. PA was higher and more stable in sodium citrate 109 mM than in 129 mM at all times. The extent of PA did not change for up to 2 hr after blood collection, and subsequently dramatically decreased. In contrast with ISTH-SSC recommendations, (1) BC does not show advantages compared to C; (2) 109 mM citrate is preferable to 129 mM, because it better supports PA; and (3) LTA studies should be completed within 2 hr of blood collection, instead of the recommended 4 hr.

  5. Feasibility Studies on Pipeline Disposal of Concentrated Copper Tailings Slurry for Waste Minimization

    NASA Astrophysics Data System (ADS)

    Senapati, Pradipta Kumar; Mishra, Barada Kanta

    2017-06-01

    The conventional lean phase copper tailings slurry disposal systems create pollution all around the disposal area through seepage and flooding of waste slurry water. In order to reduce water consumption and minimize pollution, the pipeline disposal of these waste slurries at high solids concentrations may be considered as a viable option. The paper presents the rheological and pipeline flow characteristics of copper tailings samples in the solids concentration range of 65-72 % by weight. The tailings slurry indicated non-Newtonian behaviour at these solids concentrations and the rheological data were best fitted by Bingham plastic model. The influence of solids concentration on yield stress and plastic viscosity for the copper tailings samples were discussed. Using a high concentration test loop, pipeline experiments were conducted in a 50 mm nominal bore (NB) pipe by varying the pipe flow velocity from 1.5 to 3.5 m/s. A non-Newtonian Bingham plastic pressure drop model predicted the experimental data reasonably well for the concentrated tailings slurry. The pressure drop model was used for higher size pipes and the operating conditions for pipeline disposal of concentrated copper tailings slurry in a 200 mm NB pipe with respect to specific power consumption were discussed.

  6. Feasibility Studies on Pipeline Disposal of Concentrated Copper Tailings Slurry for Waste Minimization

    NASA Astrophysics Data System (ADS)

    Senapati, Pradipta Kumar; Mishra, Barada Kanta

    2016-06-01

    The conventional lean phase copper tailings slurry disposal systems create pollution all around the disposal area through seepage and flooding of waste slurry water. In order to reduce water consumption and minimize pollution, the pipeline disposal of these waste slurries at high solids concentrations may be considered as a viable option. The paper presents the rheological and pipeline flow characteristics of copper tailings samples in the solids concentration range of 65-72 % by weight. The tailings slurry indicated non-Newtonian behaviour at these solids concentrations and the rheological data were best fitted by Bingham plastic model. The influence of solids concentration on yield stress and plastic viscosity for the copper tailings samples were discussed. Using a high concentration test loop, pipeline experiments were conducted in a 50 mm nominal bore (NB) pipe by varying the pipe flow velocity from 1.5 to 3.5 m/s. A non-Newtonian Bingham plastic pressure drop model predicted the experimental data reasonably well for the concentrated tailings slurry. The pressure drop model was used for higher size pipes and the operating conditions for pipeline disposal of concentrated copper tailings slurry in a 200 mm NB pipe with respect to specific power consumption were discussed.

  7. Feasibility of sizing metallic nanoparticles in concentrated suspensions from effective optical properties

    NASA Astrophysics Data System (ADS)

    Morales-Luna, G.; Márquez-Islas, R.; Vázquez-Estrada, O.; Contreras-Tello, H.; García-Valenzuela, A.

    2015-08-01

    We explore using measurements of the effective refractive index of a metallic nanofluid to estimate the size of the particles in it. We assume the nanofluid consists of spherical metallic nanoparticles suspended in a transparent base liquid and discuss a way of measuring the real and imaginary parts of the effective refractive index for concentrated nanofluids to about 1% in particles' volume concentration. Specifically, we consider the case of copper nanoparticles suspended in water. We propose an unambiguous effective optical parameter as a candidate to evidence the particle size, potentially in real time. Limitations due to dependent scattering effects in concentrated nanofluids are briefly stated.

  8. Use of concentrated bone marrow aspirate and platelet rich plasma during minimally invasive decompression of the femoral head in the treatment of osteonecrosis

    PubMed Central

    Martin, John R.; Houdek, Matthew T.; Sierra, Rafael J.

    2013-01-01

    The aim of this paper is to describe our surgical procedure for the treatment of osteonecrosis of the femoral head using a minimally invasive technique. We have limited the use of this procedure for patients with pre-collapse osteonecrosis of the femoral head (Ficat Stage I or II). To treat osteonecrosis of the femoral head at our institution we currently use a combination of outpatient, minimally invasive iliac crest bone marrow aspirations and blood draw combined with decompressions of the femoral head. Following the decompression of the femoral head, adult mesenchymal stem cells obtained from the iliac crest and platelet rich plasma are injected into the area of osteonecrosis. Patients are then discharged from the hospital using crutches to assist with ambulation. This novel technique was utilized on 77 hips. Sixteen hips (21%) progressed to further stages of osteonecrosis, ultimately requiring total hip replacement. Significant pain relief was reported in 86% of patients (n = 60), while the rest of patients reported little or no pain relief. There were no significant complications in any patient. We found that the use of a minimally invasive decompression augmented with concentrated bone marrow and platelet rich plasma resulted in significant pain relief and halted the progression of disease in a majority of patients. PMID:23771751

  9. Application of a portable microscopic cell counter for the counting of residual leukocytes in leukoreduced apheresis platelet concentrates in a hospital blood bank.

    PubMed

    Chun, Sejong; Kim, Eun-Young; Cha, Seung-Yeon; Seo, Ji-Young; Koo, Hong Hoe; Cho, Duck

    2017-06-01

    While a portable microscopic cell counter has been evaluated to enumerate residual white blood cells (WBCs) in red blood cells and platelet concentrates at blood centers, it has not yet been assessed in a hospital blood bank. We investigated the performance of this device and evaluated its accuracy, along with its benefits in time management. Residual WBCs from each of 100 apheresis platelet specimens were measured manually using a Nageotte chamber, along with flow cytometry methods and an ADAM-rWBC automated instrument (NanoEnTek, Seoul, South Korea). The efficiency was calculated by measuring the time required for the analysis of one specimen ten times consecutively. Flow cytometry and the ADAM-rWBC were able to detect four sporadic cases that had residual WBCs exceeding 1/μL that were not detected by the manual method. Analysis time was the shortest with the ADAM-rWBC, followed by flow cytometry and the manual method. Our data suggest that hospital blood banks require quality control of residual WBCs; among the methods evaluated in this study, the portable microscopic cell counter offers the best time efficiency. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Covalent co-immobilization of heparin/laminin complex that with different concentration ratio on titanium surface for selectively direction of platelets and vascular cells behavior

    NASA Astrophysics Data System (ADS)

    Wang, Jian; Chen, Yuan; Liu, Tao; Wang, Xue; Liu, Yang; Wang, Yuan; Chen, Junying; Huang, Nan

    2014-10-01

    Surface biofunctional modification of coronary artery stent to improve the hemocompatibility and selectively accelerate endothelium regeneration but prevent restenosis have been become a new hotspot. For this, a novel method was developed in this work by co-immobilization of Ln and heparin complex on poly-L-lysine modified Ti surface. Take the advantage of the specific interaction between Ln and heparin, Ln and heparin complexes with different concentration ratios were set up for creating different exposure density of these two types of biomolecules. According to biocompatibility evaluation results, the Hep/Ln complexes modified surface displayed less platelet adhesion and activation. Especially, on L(150)H and L(200)H surface, the AT III binding quantity, APTT value and anti-coagulation property of modified surface were significantly promoted. Furthermore, the adherent density and proliferation activity of ECs and EPCs were positively correlated with Ln concentration. Notably, the proliferation of both ECs and EPCs on L(100)H, L(150)H and L(200)H surface were greatly promoted. Another hand, the proliferation activity of SMCs was significantly inhibited on Hep/Ln modified surfaces, which was considered mainly due to the inhibitory effect of heparin to SMCs. According to the existing results, this study demonstrated that in a certain range of heparin and laminin concentration ratio, the biological behavior of platelets, ECs, EPCs and SMCs could be selectively directed. We suggested that this article provided a potential method to construct an adequate platform on a stent surface for accelerate endothelialization with low side effects.

  11. Stability of buffer-free freeze-dried formulations: A feasibility study of a monoclonal antibody at high protein concentrations.

    PubMed

    Garidel, Patrick; Pevestorf, Benjamin; Bahrenburg, Sven

    2015-11-01

    We studied the stability of freeze-dried therapeutic protein formulations over a range of initial concentrations (from 40 to 160 mg/mL) and employed a variety of formulation strategies (including buffer-free freeze dried formulations, or BF-FDF). Highly concentrated, buffer-free liquid formulations of therapeutic monoclonal antibodies (mAbs) have been shown to be a viable alternative to conventionally buffered preparations. We considered whether it is feasible to use the buffer-free strategy in freeze-dried formulations, as an answer to some of the known drawbacks of conventional buffers. We therefore conducted an accelerated stability study (24 weeks at 40 °C) to assess the feasibility of stabilizing freeze-dried formulations without "classical" buffer components. Factors monitored included pH stability, protein integrity, and protein aggregation. Because the protein solutions are inherently self-buffering, and the system's buffer capacity scales with protein concentration, we included highly concentrated buffer-free freeze-dried formulations in the study. The tested formulations ranged from "fully formulated" (containing both conventional buffer and disaccharide stabilizers) to "buffer-free" (including formulations with only disaccharide lyoprotectant stabilizers) to "excipient-free" (with neither added buffers nor stabilizers). We evaluated the impacts of varying concentrations, buffering schemes, pHs, and lyoprotectant additives. At the end of 24 weeks, no change in pH was observed in any of the buffer-free formulations. Unbuffered formulations were found to have shorter reconstitution times and lower opalescence than buffered formulations. Protein stability was assessed by visual inspection, sub-visible particle analysis, protein monomer content, charge variants analysis, and hydrophobic interaction chromatography. All of these measures found the stability of buffer-free formulations that included a disaccharide stabilizer comparable to buffer

  12. Platelet fibrinogen

    PubMed Central

    Castaldi, P. A.; Caen, J.

    1965-01-01

    Platelet fibrinogen has been studied in normal, thrombasthenic, and hypofibrinogenaemic subjects. It has been differentiated into adsorbed (plasma) and extractable (intraplatelet) fractions. Isotopic studies suggest that exchange does not occur between intraplatelet and plasma fibrinogen and it appears possible that the intra-platelet fraction may be derived from the megakaryocyte. Six of nine thrombasthenic patients were found to have a severe deficiency of both adsorbed and extractable fibrinogen. Since the remaining three had near-normal platelet fibrinogen and all nine failed to aggregate it is improbable that the failure to adsorb fibrinogen is responsible for the defect in aggregation. Magnesium partially corrects adhesion to fibrin and clot retraction by these platelets, but has not been found to influence their fibrinogen adsorption. It is considered that the basic platelet surface defect, of varying severity, is responsible for the abnormalities of adsorption, aggregation, and adhesion in thrombasthenia. In the case of congenital hypofibrinogenaemia, fibrinogen transfusion corrects the long bleeding time, platelet-adsorbed fibrinogen, and the ability of platelets to spread on glass. It is possible that fibrinogen influences the surface properties of human platelets, although the final mechanism is not determined. Images PMID:5835438

  13. Olanzapine and risperidone plasma concentration therapeutic drug monitoring: A feasibility study.

    PubMed

    Law, Suzanne; Gudbrandsen, Maria; Magill, Nicholas; Sweetman, Isabel; Rose, Diana; Landau, Sabine; Flanagan, Robert J; David, Anthony S; Patel, Maxine X

    2015-08-01

    This study aimed to develop a clinically acceptable method of therapeutic drug monitoring (TDM) for olanzapine and risperidone and to evaluate the feasibility of its implementation. A non-randomised study of inpatients from five Mental Health Trusts was conducted, with a clinical interview at the time of TDM and a subsequent 6-week follow-up review of clinical notes. The TDM intervention comprised: (a) a venous blood sample taken 12 hours post-dose, 7-10 days after drug initiation, and (b) rapid results feedback, with interpretation algorithm guidance. Thirty-two participants provided samples (19 prescribed olanzapine, 13 risperidone). Twenty-six participants remained on the target drug at study end, with seven experiencing a dose change, for whom only four of the TDM results were confirmed as having been checked. Mean dose increased for olanzapine (0.9 mg/day, range 0-10) and decreased for risperidone (-0.3 mg/day, range -4-3). TDM can be implemented as part of routine clinical practice for both drugs. However, the lack of robust supporting evidence for or against antipsychotic TDM has probably led to a lack of enthusiasm for and interest in the results. Nevertheless, the advent of less invasive measures and the targeting of patients who might be more likely to benefit may facilitate uptake. © The Author(s) 2015.

  14. Nowcasting and Forecasting Concentrations of Biological Contaminants at Beaches: A Feasibility and Case Study

    EPA Science Inventory

    Public concern over microbial contamination of recreational waters has increased in recent years. A common approach to evaluating beach water quality has been to use the persistence model which assumes that day-old monitoring results provide accurate estimates of current concentr...

  15. The Feasibility of a Stretched Lens Concentrating Solar Array Direct-Driving an Electric Thruster

    NASA Astrophysics Data System (ADS)

    Brandhorst, Henry W.; Best, Steve R.; Rodiek, Julie A.

    2010-01-01

    As space exploration continues to be a primary focus of NASA, solar electric propulsion (SEP) becomes a forerunner in the mode of transportation to reach the moon and other planets in our solar system. The Stretched Lens Array (SLA) is a unique ultra-high-performance, ultra-light, cost-effective photovoltaic concentrator array using refractive concentrator technology. The SLA is capable of high voltage operation and sustainability in a high radiation environment and can be specifically optimized for SEP by the ability to direct-drive Hall-effect thrusters. Auburn University has performed a ``direct drive'' experiment using a high-voltage (600 Voc) ENTECH SunLine concentrator array powered with multijunction solar cells coupled to a Russian T-100 Hall Effect Thruster (HET). This appears to be the first time a Hall thruster has been run directly from III-V-based multi-junction solar cells and at this high voltage. This paper discusses the set-up and testing results. Testing includes the inclusion of ENTECH's Stretched Lens Array hardware in a vacuum chamber to measure plume impingement effects at various positions relative to the exhaust axis of the thruster. The goal is to define meaningful high voltage SLA concentrator array and Hall thruster demonstration tests relevant to SEP and to test SLA reliability.

  16. Economic feasibility of segregating dark northern spring wheat by protein concentration during harvest

    USDA-ARS?s Scientific Manuscript database

    In-line, optical sensing has been developed for on-combine measurement and mapping of grain protein concentration (GPC). The objective of this study was to estimate changes in costs and net returns from using this technology for segregation of the dark northern spring (DNS) subclass of hard red whe...

  17. Nowcasting and Forecasting Concentrations of Biological Contaminants at Beaches: A Feasibility and Case Study

    EPA Science Inventory

    Public concern over microbial contamination of recreational waters has increased in recent years. A common approach to evaluating beach water quality has been to use the persistence model which assumes that day-old monitoring results provide accurate estimates of current concentr...

  18. Haemostasis monitored in stored red blood cells, plasma and platelet concentrates in the proportion of 4 :  4 :  1 diluted with crystalloids and colloids.

    PubMed

    Ågren, Anna; Edgren, Gustaf; Ambrosio, Daniela; Gryfelt, Gunilla; Östlund, Anders; Wikman, Agneta

    2016-04-01

    The aim of this in-vitro study was to evaluate haemostasis analysed with thromboelastometry and blood gas and blood count variables, in stored blood components and the effects after dilution with Ringer[Combining Acute Accent]s acetate, albumin and hydroxyethyl starch (HES). Aliquots from stored red blood cells, plasma and platelet concentrates were mixed in the proportion of 4 : 4 : 1 and analysed with rotational thromboelastometry (ROTEM), blood count [haemoglobin (Hb), haematocrit, platelet count] and blood gas (pH, calcium, sodium, potassium, glucose levels). The blood mix was thereafter diluted 20 and 33% with Ringer's acetate, albumin or HES. The stored blood component mix in a ratio of 4 : 4 : 1 had a low pH (7.11 ± 0.03, mean ± standard deviation), nonmeasurable calcium level, and high concentrations of sodium, potassium and glucose but ROTEM curves within normal range after recalcification. With Ringer's acetate dilution, the ROTEM variables changed almost linearly with increasing dilution volume. When albumin was used in the 33% dilution, the clot firmness of the fibrin clot (FibTEM) was further reduced, and with HES dilution, there was a pronounced impairment. The stored blood mix had a low pH and calcium level, both of which might have a significant influence on the coagulation process but normal ROTEM curves after recalcification. Dilution with Ringer's acetate and albumin resulted in moderate deterioration, while dilution with HES showed severely impaired haemostasis.

  19. Establishment of a proficiency panel for an external quality assessment programme for the detection of bacterial contamination in platelet concentrates using rapid and cultural detection methods.

    PubMed

    Vollmer, T; Schmidt, M; Hourfar, K; Schottstedt, V; Pichl, L; Gubbe, K; Knabbe, C; Dreier, J

    2016-05-01

    Platelet concentrates (PCs) are the main focus regarding the residual risk of transfusion-transmitted bacterial infections. Rapid screening methods for bacterial detection in platelets have been optimized over the last decade, but their external evaluation represents a complicated process. We developed a new type of proficiency panel for bacterial detection in PCs using currently available screening methods (especially rapid methods) suitable for external quality assessment programmes (EQAP). PC samples were inoculated with different bacteria at two concentrations (10E+03 CFU/ml, 10E+05 CFU/ml) and stored under temperature-controlled conditions (1-5 days). Bacterial growth was further prevented by the addition of 0-20 μg/ml cotrimoxazole. Samples were analysed prior to and after storage using rapid detection methods (Bactiflow (BF), bacteria-generic NAT) and cultural methods to determine the influence of storage and antibiotic treatment on bacterial counts and the result outcome. A pilot EQAP was performed with four participants. Testing under the evaluated conditions demonstrated that bacterial counts remained constant prior to and after storage. The supplementation of 10 μg/ml cotrimoxazole did not influence bacterial detection using the two rapid detection methods BF and NAT. Furthermore, the detection of bacteria using cultural methods is still possible despite of antibiotic supplementation. The pilot EQAP confirmed these results. A storage time of up to 3 days proved practicable, showing no considerable influence on bacterial count and outcome of test results. The established proficiency panel provided PC matrix-conform samples with stabilized bacterial counts which can be analysed in parallel by rapid and cultural detection methods. © 2016 International Society of Blood Transfusion.

  20. Effect of ASA dose doubling versus switching to clopidogrel on plasma inflammatory markers concentration in patients with type 2 diabetes and high platelet reactivity: the AVOCADO study.

    PubMed

    Rosiak, Marek; Postula, Marek; Kaplon-Cieslicka, Agnieszka; Kondracka, Agnieszka; Trzepla, Ewa; Czlonkowski, Andrzej; Janicki, Piotr K; Filipiak, Krzysztof J; Opolski, Grzegorz

    2013-01-01

    The aim of the study was to compare the effects of 2 strategies of antiplatelet treatment (i.e., 150 mg ASA vs. 75 mg clpoidogrel) on plasma level of inflammatory markers in type 2 diabetes mellitus (T2DM) patients with high platelet reactivity (HPR). Study cohort consisted of 304 T2DM patients on chronic ASA therapy (75 mg per day) participating in the Aspirin Versus/Or Clopidogrel in Aspirin-resistant Diabetics inflammation Outcomes (AVOCADO) study. Patients with HPR defined as Platelet Function Analyzer (PFA)-100 collagene/epinephrine closure time (CEPI-CT) < 193 s (n = 80) were randomized to 150 mg of ASA or 75 mg of clopidogrel in 2:3 ratio, respectively. Concentrations of the selected inflammatory markers, including tumor necrosis factor (TNF)-α, interleukin (IL)-6, solubleCD40 ligand (sCD40L), and high sensitivity C-reactive protein (hsCRP), were measured and compared in both treatment groups before and after 8 weeks of treatment in both groups. Out of 234 patients included into final analysis, the total of 34.2% (n = 80) patients displayed HPR, of which 14.1% (n = 33) were randomized into 150 mg of ASA group and 20.1% (n = 47) into 75 mg of clopidogrel group. Treatment with clopidogrel was a positive predictor (stepwise multiple regression analysis) of reduction of sCD40L concentration (odds ratio [OR] 4.15; p = 0.013), while treatment with 150 mg ASA was a positive predictor of reduction of IL-6 concentration (OR 4.38; p = 0.033). There was no statistically significant differences between clopidogrel and ASA 150 mg treatment in respect to predictive value for decreased hsCRP concentrations or increased TNF-α concentrations. Increasing the dose of ASA from 75 mg to 150 mg daily or switching ASA 75 mg to clopidogrel 75 mg daily may reduce concentrations of some inflammatory markers (in particular hsCRP, IL-6 and CD40L) in T2DM patients with HPR treated previously with 75 mg of ASA.

  1. Feasibility of Estimating Constituent Concentrations and Loads Based on Data Recorded by Acoustic Instrumentation

    USGS Publications Warehouse

    Lietz, A.C.

    2002-01-01

    The acoustic Doppler current profiler (ADCP) and acoustic Doppler velocity meter (ADVM) were used to estimate constituent concentrations and loads at a sampling site along the Hendry-Collier County boundary in southwestern Florida. The sampling site is strategically placed within a highly managed canal system that exhibits low and rapidly changing water conditions. With the ADCP and ADVM, flow can be gaged more accurately rather than by conventional field-data collection methods. An ADVM velocity rating relates measured velocity determined by the ADCP (dependent variable) with the ADVM velocity (independent variable) by means of regression analysis techniques. The coefficient of determination (R2) for this rating is 0.99 at the sampling site. Concentrations and loads of total phosphorus, total Kjeldahl nitrogen, and total nitrogen (dependent variables) were related to instantaneous discharge, acoustic backscatter, stage, or water temperature (independent variables) recorded at the time of sampling. Only positive discharges were used for this analysis. Discharges less than 100 cubic feet per second generally are considered inaccurate (probably as a result of acoustic ray bending and vertical temperature gradients in the water column). Of the concentration models, only total phosphorus was statistically significant at the 95-percent confidence level (p-value less than 0.05). Total phosphorus had an adjusted R2 of 0.93, indicating most of the variation in the concentration can be explained by the discharge. All of the load models for total phosphorus, total Kjeldahl nitrogen, and total nitrogen were statistically significant. Most of the variation in load can be explained by the discharge as reflected in the adjusted R2 for total phosphorus (0.98), total Kjeldahl nitrogen (0.99), and total nitrogen (0.99).

  2. Feasibility of Topical Applications of Natural High-Concentration Capsaicinoid Solutions in Patients with Peripheral Neuropathic Pain: A Retrospective Analysis

    PubMed Central

    Mouraux, Andre; Deumens, Ronald; Leerink, Marjolein; le Polain de Waroux, Bernard; Joëlle, Quetin-Leclercq

    2016-01-01

    Background. Capsaicin, one of several capsaicinoid compounds, is a potent TRPV1 agonist. Topical application at high concentration (high concentration, >1%) induces a reversible disappearance of epidermal free nerve endings and is used to treat peripheral neuropathic pain (PNP). While the benefit of low-concentration capsaicin remains controversial, the 8%-capsaicin patch (Qutenza®, 2010, Astellas, Netherlands) has shown its effectiveness. This patch is, however, costly and natural high-concentration capsaicinoid solutions may represent a cheaper alternative to pure capsaicin. Methods. In this retrospective study, 149 patients were screened, 132 were included with a diagnosis of neuropathic pain, and eighty-four were retained in the final analyses (median age: 57.5 years [IQR25–75: 44.7–67.1], male/female: 30/54) with PNP who were treated with topical applications of natural high-concentration capsaicinoid solutions (total number of applications: 137). Indications were postsurgical PNP (85.7%) and nonsurgical PNP (14.3%) (posttraumatic, HIV-related, postherpetic, and radicular PNP). Objectives. To assess the feasibility of topical applications of natural high-concentration capsaicinoid solutions for the treatment of PNP. Results. The median treated area was 250 cm2 [IQR25–75: 144–531]. The median amount of capsaicinoids was 55.1 mg [IQR25–75: 28.7–76.5] per plaster and the median concentration was 172.3 μg/cm2 [IQR25–75: 127.6–255.2]. Most patients had local adverse effects on the day of treatment, such as mild to moderate burning pain and erythema. 13.6–19.4% of the patients experienced severe pain or erythema. Following treatment, 62.5% of patients reported a lower pain intensity or a smaller pain surface, and 35% reported a sustained pain relief lasting for at least 4 weeks. Conclusion. Analgesic topical treatment with natural high-concentration capsaicinoid is feasible and may represent a low cost alternative to alleviate PNP in

  3. Current status of additive solutions for platelets.

    PubMed

    Alhumaidan, Hiba; Sweeney, Joseph

    2012-01-01

    The storage of platelets in additive solution (PAS) had lagged behind red cell concentrates, especially in North America. The partial or complete removal of anticoagulated plasma and storage of platelet concentrates in AS presents many advantages. The PAS can be formulated to optimize aerobic metabolism or decrease platelet activation, thus abrogating the platelet storage lesion and potentially improving in vivo viability. Plasma removal has been shown to reduce allergic reactions and the plasma harvested could contribute to the available plasma pool for transfusion or fractionation. PAS coupled to pathogen reduction technology results in a platelet product of equivalent hemostatic efficacy to conventionally stored platelets. Given the above, the likely future direction of platelet storage will be in new generation designer PAS with an extended shelf life and a superior safety profile to plasma stored platelets. J. Clin. Apheresis, 2012. © 2012 Wiley Periodicals, Inc.

  4. [Effects of riboflavin combined with photosensitization on reduction of Gram-positive and Gram-negative indicating germs in plasma and P-selectin expression of apheresis platelet concentrates].

    PubMed

    Zhou, Xue-Yin; Xiong, Wen; Kong, Ling-Kui

    2010-08-01

    This study was purposed to confirm the practical efficacy of reducing indicating germs suspended in plasma by riboflavin and photosensitized inactivation and to evaluate its influence on activation of apheresis platelet concentrates. The synergistic effects of riboflavin combined with ultraviolet irradiation on inactivation of germs were investigated by using Escherichia Coli (E. coli) and Staphylococcus Aureus (S. aureus) as Gram⁻ and Gram(+) indicating germs, respectively. The activation status of apheresis-platelet concentrates treated with riboflavin combined with ultraviolet irradiation was detected by flow cytometry. The results showed that when 50 μmol/L of riboflavin was combined with 6.2 J/ml of ultraviolet irradiation, the T/E ratios reached 1.42 for E. coli and 1.68 for S. Aureus, and reduction of E. Coli and S. Aureus were 3.87 Logs and 3.82 Logs respectively; the CD62p expression level on germ-inactivated platelets stored at 22 degrees C for 0 and 5 days were 4.92% and 36.18% respectively, which slightly increased as compared with controls (3.94% and 32.03)% (p < 0.05). It is concluded that combination of riboflavin with ultraviolet irradiation displays well synergistic effects which can reduce E. Coli and S. Aureus counts, but no significantly influence on platelets. The partial activation of liquid platelets mainly presents metabolism damage during storage, which is found at an acceptable level.

  5. EFFECTS OF BACTERIAL ENDOTOXIN ON RABBIT PLATELETS

    PubMed Central

    Des Prez, Roger M.; Bryant, Richard E.

    1966-01-01

    The divalent ion requirements of rabbit platelet injury by endotoxin have been defined by the use of various anticoagulant solutions and have been compared to the divalent ion requirements of platelet injury produced by addition of antigen to immune platelet-rich plasma. The endotoxin-platelet interaction takes place in citrated blood. Platelet damage by antigen is inhibited by citrate, but preincubation of antigen and immune platelet-poor plasma in the absence of citrate results in a substance, presumably antigen-antibody complement complex, which then does injure platelets in the presence of citrate. Neither endotoxin nor preincubated antigen injures platelets in the presence of sodium EDTA in concentrations sufficient to interact with all divalent cations present in plasma. These observations have been interpreted by viewing the platelet-endotoxin interaction as a consequence of platelet phagocytosis of endotoxin, a reaction not requiring complement but requiring definite small concentrations of divalent cations. The interaction of antigen and platelets is regarded as a two phase reaction, the first requiring the participation of complement and concentrations of divalent cation larger than those provided in citrated plasma, the second requiring smaller concentrations of divalent cation, no further participation of complement, and active in citrated plasma. This second phase is regarded as representing platelet phagocytosis of immune complexes. PMID:5951281

  6. Biologic nanoparticles and platelet reactivity

    PubMed Central

    Miller, Virginia M; Hunter, Larry W; Chu, Kevin; Kaul, Vivasvat; Squillace, Phillip D; Lieske, John C; Jayachandran, Muthuvel

    2009-01-01

    Aim Nanosized particles (NPs) enriched in hydroxyapatite and protein isolated from calcified human tissue accelerate occlusion of endothelium-denuded arteries when injected intravenously into rabbits. Since platelet aggregation and secretory processes participate in normal hemostasis, thrombosis and vascular remodeling, experiments were designed to determine if these biologic NPs alter specific platelet functions in vitro. Methods Platelet-rich plasma was prepared from citrate anticoagulated human blood. Platelet aggregation and ATP secretion were monitored in response to thrombin receptor agonists peptide (10 μM) or convulxin (50 μg/ml) prior to and following 15 min incubation with either control solution, human-derived NPs, bovine-derived NPs or crystals of hydroxyapatite at concentrations of 50 and 150 nephelometric turbidity units. Results Incubation of platelets for 15 min with either human- or bovine-derived NPs reduced aggregation induced by thrombin receptor activator peptide and convulxin in a concentration-dependent manner. Hydroxyapatite caused a greater inhibition than either of the biologically derived NPs. Human-derived NPs increased ATP secretion by unstimulated platelets during the 15 min incubation period. Conclusion Effects of bovine-derived and hydroxyapatite NPs on basal release of ATP were both time and concentration dependent. These results suggest that biologic NPs modulate both platelet aggregation and secretion. Biologically derived NPs could modify platelet responses within the vasculature, thereby reducing blood coagulability and the vascular response to injury. PMID:19839809

  7. Feasibility of hydroxyl concentration measurements by laser-saturated fluorescence in high-pressure flames

    NASA Technical Reports Server (NTRS)

    Carter, Campbell D.; King, Galen B.; Laurendeau, Normand M.; Salmon, J. Thaddeus

    1987-01-01

    The effect of pressure on the laser-saturated fluorescence method for measuring OH concentration in high-pressure flames is studied using calculations for the burned-gas region of a stoichiometric H2-O2 flame at 2000 K. A numerical model of the excitation dynamics of OH is developed to explore the validity of the balanced cross-rate model at higher pressures. It is shown that depopulation of the laser-coupled levels is sensitive to collisions which depopulate v-double-prime (VDP) = 0 and to rate coefficients for rotational transfer in the ground state which are smaller than those in the excited state. In particular, it is shown that the depopulation of VDP = 0, and hence the laser-coupled levels, depends on the probability of electronic quenching to vibrational levels for which VDP is greater than 0 and vibrational relaxation to VDP = 0.

  8. Analysis of a concentric-tube robot design and feasibility for endoscopic deployment

    NASA Astrophysics Data System (ADS)

    Ponten, Ryan; Black, Caroline B.; Russ, Andrew J.; Rucker, D. Caleb

    2017-03-01

    An intraluminal endoscopic approach is desirable for most colonoscopic procedures and is growing in favor for other surgeries as tools are enhanced. Flexible robotic manipulators could further enhance the dexterity and precision of commercial endoscopic systems. In this paper, we explore the capabilities of concentric tube robots to work as tool manipulators at the tip of a colonoscope to perform endoscopic submucousal dissection (ESD) and endoscopic full thickness resection (EFTR). We provide an overview of the kinematic modeling of these manipulators, a design of a prototype manipulator and the transmission actuation system. Our analysis examines the workspace and stiffness of these manipulators being controlled at the tip of a colonoscope. We compare the results to reported surgical requirements and propose solutions for enhancing their effectiveness including notching tubes with a larger Young's Modulus. We also determine the resolution and accuracy of the actuation system.

  9. PLATELET FORMATION

    PubMed Central

    Thon, Jonathan N.; Italiano, Joseph E.

    2010-01-01

    Thrombocytopenia is the underlying cause of a number of major clinical conditions and genetic disorders worldwide. While therapeutic agents that bind and stimulate the thrombopoietin receptor are currently available, the development of drugs that directly stimulate megakaryocytes to generate platelets has lagged behind. To improve the management of thrombocytopenia, we will need to define the cell biological pathways that drive the production of platelets from megakaryocytes. This review integrates the latest research of platelet biogenesis and focuses on the molecular pathways that power and regulate proplatelet production. PMID:20620432

  10. Feasibility of antibody-poly(glutamic acid) complexes: preparation of high-concentration antibody formulations and their pharmaceutical properties.

    PubMed

    Izaki, Shunsuke; Kurinomaru, Takaaki; Maruyama, Takuya; Uchida, Takayuki; Handa, Kenji; Kimoto, Tomoaki; Shiraki, Kentaro

    2015-06-01

    Development of high-concentration antibody formulations for subcutaneous administration remains challenging. Recently, a precipitation-redissolution method was proposed to prepare suspensions or precipitates of salt-dissociable protein-poly(amino acid) complexes. To elucidate the utility of this method for protein therapy, we investigated the feasibility of a precipitation-redissolution method using poly(amino acid) for high-concentration antibody formulation. Omalizumab and adalimumab formulations of 150 mg/mL could be prepared using poly-l-glutamic acid (polyE) from low-concentration stock solutions. Enzyme-linked immunosorbent assay, circular dichroism, and size-exclusion chromatography revealed that the formation of antibody-polyE complex and precipitation-redissolution process did not significantly affect the immunoreactivity or secondary structure of the antibodies. The precipitation-redissolution method was less time-consuming and more effective than lyophilization-redissolution, evaporation-redissolution, and ultrafiltration from the viewpoint of final yield. Scalability was confirmed from 400 μL to 1.0 L. The general toxicity and pharmacokinetic profiles of the antibody-polyE complex formulations were similar to those of conventional antibody formulations. These results suggested that the precipitation-redissolution method using poly(amino acid) has great potential as a concentration method for antibody formulation and medicinal use.

  11. Osmotic stability of blood platelets.

    PubMed

    Fantl, P

    1968-09-01

    1. Hypotonic solutions added to human platelet-containing plasma cause a transient decrease of absorbancy of light at 610 mmu which is followed by a gradual increase of absorbancy.2. When platelets are stored for 7 hr at 4 degrees C the absorbancy changes with variations of osmolarity and their aggregation with adenosine diphosphate (ADP) remain the same. However, the reversal of absorbancy declines during storage of platelet-containing plasma.3. Platelets are not aggregated by stearate. Platelets appear to be only slightly affected by stearate concentration higher than 0.8 mM, but oleate has no effect.4. Hypertonic solutions of NaCl and urea cause increase in absorbancy of platelet-containing human plasma. Hypertonic sucrose solutions produce no more change than isotonic solutions. Hypertonic NaCl produces permanent increases in absorbancy. In human platelet-containing plasma the increased absorbancy caused by hypertonic urea is transient and declines.5. The osmotic platelet changes occur in isolated platelets as well as in platelet-containing plasma.6. The absorbancy of frozen and thawed platelet-containing plasma is not significantly altered by hypotonic solutions but the absorbancy changes caused by hypertonic solutions are similar to that of unfrozen plasma.7. The immediate absorbancy changes caused by hypo- and by hypertonic solutions are the same at 5 degrees C and 30 degrees C and are therefore probably of a physical nature. The reversal of absorbancy and aggregation of platelets by added adenosine diphosphate have Q(10) > 1 and are therefore probably of a chemical-enzymic nature.8. Divalent cations and contact activation are not required for the osmotic platelet changes and 10(-3)M-Cu(2+) and Zn(2+) do not interfere. Inhibitors of oxidative phosphorylation, electron transfer, sodium, potassium activated adenosine triphosphatases and adenosine triphosphate do not inhibit reversal of absorbancy of platelets exposed to hypotonic solutions. Cyanide, 5 x 10

  12. Osmotic stability of blood platelets

    PubMed Central

    Fantl, P.

    1968-01-01

    1. Hypotonic solutions added to human platelet-containing plasma cause a transient decrease of absorbancy of light at 610 mμ which is followed by a gradual increase of absorbancy. 2. When platelets are stored for 7 hr at 4° C the absorbancy changes with variations of osmolarity and their aggregation with adenosine diphosphate (ADP) remain the same. However, the reversal of absorbancy declines during storage of platelet-containing plasma. 3. Platelets are not aggregated by stearate. Platelets appear to be only slightly affected by stearate concentration higher than 0·8 mM, but oleate has no effect. 4. Hypertonic solutions of NaCl and urea cause increase in absorbancy of platelet-containing human plasma. Hypertonic sucrose solutions produce no more change than isotonic solutions. Hypertonic NaCl produces permanent increases in absorbancy. In human platelet-containing plasma the increased absorbancy caused by hypertonic urea is transient and declines. 5. The osmotic platelet changes occur in isolated platelets as well as in platelet-containing plasma. 6. The absorbancy of frozen and thawed platelet-containing plasma is not significantly altered by hypotonic solutions but the absorbancy changes caused by hypertonic solutions are similar to that of unfrozen plasma. 7. The immediate absorbancy changes caused by hypo- and by hypertonic solutions are the same at 5° C and 30° C and are therefore probably of a physical nature. The reversal of absorbancy and aggregation of platelets by added adenosine diphosphate have Q10 > 1 and are therefore probably of a chemical-enzymic nature. 8. Divalent cations and contact activation are not required for the osmotic platelet changes and 10-3 M-Cu2+ and Zn2+ do not interfere. Inhibitors of oxidative phosphorylation, electron transfer, sodium, potassium activated adenosine triphosphatases and adenosine triphosphate do not inhibit reversal of absorbancy of platelets exposed to hypotonic solutions. Cyanide, 5 × 10-3 M, fluoride, 1

  13. Bench Test for the Detection of Bacterial Contamination in Platelet Concentrates Using Rapid and Cultural Detection Methods with a Standardized Proficiency Panel.

    PubMed

    Vollmer, Tanja; Knabbe, Cornelius; Geilenkeuser, Wolf-Jochen; Schmidt, Michael; Dreier, Jens

    2015-07-01

    The most frequent infectious complication in transfusion therapy in developed countries is related to the bacterial contamination of platelet concentrates (PCs). Rapid and cultural screening methods for bacterial detection in platelets are available, but external performance evaluation, especially of rapid methods, has been difficult to realize so far. Here we summarize the results of three individual collaborative trials using an external quality assessment program (EQAP) for the application of current rapid and cultural screening methods. Three different modules were available for the detection of bacterial contamination: module 1: rapid methods, module 2: culture methods, module 3: bacterial identification methods. The sample set-up included up to six different bacterial strains, 1-2 negative samples and 4-6 positive samples with stabilized bacterial cell counts (approximately 10(3)/10(4)/10(5) CFU/ml). Time schedule for testing was limited (module 1: 6 h, module 2 and 3: 7 days). Samples of module 1 were analyzed with two different rapid methods (BactiFlow, NAT). The results of the three individual collaborative trials showed that all participants detected the negative samples with both assays correctly. Samples spiked with 10(4) to 10(5) CFU/ml of bacteria obtained positive results with both rapid screening methods, whereas samples spiked with only 10(3) CFU/ml disclosed a lower number of correctly identified positive results by NAT (86.6-93.8% sensitivity) compared to BactiFlow (100% sensitivity). The results for modules 2 and 3 revealed a 100% diagnostic sensitivity and specificity in all three collaborative trials. This proficiency panel facilitates the verification of the analytical sensitivity of rapid and cultural bacterial detection systems under controlled routine conditions. The concept of samples provided in this EQAP has three main advantages: i) samples can be examined by both rapid and culture methods, ii) the provided material is matrix

  14. Efficiency of riboflavin and ultraviolet light treatment against high levels of biofilm-derived Staphylococcus epidermidis in buffy coat platelet concentrates.

    PubMed

    Taha, M; Culibrk, B; Kalab, M; Schubert, P; Yi, Q-L; Goodrich, R; Ramirez-Arcos, S

    2017-07-01

    Staphylococcus epidermidis forms surface-attached aggregates (biofilms) in platelet concentrates (PCs), which are linked to missed detection during PC screening. This study was aimed at evaluating the efficacy of riboflavin-UV treatment to inactivate S. epidermidis biofilms in buffy coat (BC) PCs. Biofilm and non-biofilm cells from S. epidermidis ST-10002 and S. epidermidis AZ-66 were individually inoculated into whole blood (WB) units (~10(6) colony-forming units (CFU)/ml) (N = 4-5). One spiked and three unspiked WB units were processed to produce a BC-PC pool. Riboflavin was added to the pool which was then split into two bags: one for UV treatment and the second was untreated. Bacterial counts were determined before and after treatment. In vitro PC quality was assessed by flow cytometry and dynamic light scattering. Bacterial counts were reduced during BC-PC production from ~10(6) CFU/ml in WB to 10(3) -10(4) CFU/ml in PCs (P < 0·0001). Riboflavin-UV treatment resulted in significantly higher reduction of S. epidermidis AZ-66 than strain ST-10002 (≥3·5 log reduction and 2·6-2·8 log reduction, respectively, P < 0·0001). Remaining bacteria post-treatment were able to proliferate in PCs. No differences in S. epidermidis inactivation were observed in PCs produced from WB inoculated with biofilm or non-biofilm cells (P > 0·05). Platelet activation was enhanced in PCs produced with WB inoculated with biofilms compared to non-biofilm cells (P < 0·05). Riboflavin-UV treatment was similarly efficacious in PCs produced from WB inoculated with S. epidermidis biofilm or non-biofilm cells. Levels of biofilm-derived S. epidermidis ≥10(3) CFU/ml were not completely inactivated; however, further testing is necessary with lower (real-life) bacterial levels. © 2017 International Society of Blood Transfusion.

  15. Concentrating-collector mass-production feasibility. Volume I. Final report

    SciTech Connect

    Not Available

    1981-11-02

    The Performance Prototype Trough (PPT) Concentrating Collector consists of four 80-foot modules in a 320-foot row. The collector was analyzed, including cost estimates and manufacturing processes to produce collectors in volumes from 100 to 100,000 modules per year. The four different reflector concepts considered were the sandwich reflector structure, sheet metal reflector structure, molded reflector structure, and glass laminate structure. The sheet metal and glass laminate structures are emphasized with their related structure concepts. A preliminary manufacturing plan is offered that includes: documentation of the manufacturing process with production flow diagrams; labor and material costs at various production levels; machinery and equipment requirements including preliminary design specifications; and capital investment costs for a new plant. Of five reflector designs considered, the two judged best and considered at length are thin annealed glass and steel laminate on steel frame panel and thermally sagged glass. Also discussed are market considerations, costing and selling price estimates, design cost analysis and make/buy analysis. (LEW)

  16. Platelets actively sequester angiogenesis regulators

    PubMed Central

    Yip, Tai-Tung; Cassiola, Flavia; Kikuchi, Lena; Cervi, David; Podust, Vladimir; Italiano, Joseph E.; Wheatley, Erin; Abou-Slaybi, Abdo; Bender, Elise; Almog, Nava; Kieran, Mark W.; Folkman, Judah

    2009-01-01

    Clinical trials with antiangiogenic agents have not been able to validate plasma or serum levels of angiogenesis regulators as reliable markers of cancer presence or therapeutic response. We recently reported that platelets contain numerous proteins that regulate angiogenesis. We now show that accumulation of angiogenesis regulators in platelets of animals bearing malignant tumors exceeds significantly their concentration in plasma or serum, as well as their levels in platelets from non–tumor-bearing animals. This process is selective, as platelets do not take up a proportional amount of other plasma proteins (eg, albumin), even though these may be present at higher concentrations. We also find that VEGF-enriched Matrigel pellets implanted subcutaneously into mice or the minute quantities of VEGF secreted by microscopic subcutaneous tumors (0.5-1 mm3) result in an elevation of VEGF levels in platelets, without any changes in its plasma levels. The profile of other angiogenesis regulatory proteins (eg, platelet-derived growth factor, basic fibroblast growth factor) sequestered by platelets also reflects the presence of tumors in vivo before they can be macroscopically evident. The ability of platelets to selectively take up angiogenesis regulators in cancer-bearing hosts may have implications for the diagnosis and management of many angiogenesis-related diseases and provide a guide for antiangiogenic therapies. PMID:19036702

  17. The feasibility of concentrated rural settlement in a context of post-disaster reconstruction: a study of China.

    PubMed

    Peng, Yi; Shen, Liyin; Zhang, Xiaoling; Ochoa, J Jorge

    2014-01-01

    There is growing appreciation of the use of concentrated rural settlement as an effective means of implementing infrastructure projects and helping to achieve sustainable development in rural areas. This occurs in China through the exchange of rural residential land for urban construction. However, this policy has not been effective under normal circumstances (called development-driven conditions) as frequently farmers are reluctant to accept such an exchange. By contrast, in a time of disaster, such as after the 2008 earthquake in Sichuan Province, China, rural victims have accepted this policy of rural residential land exchange. Employing game theory, this paper identifies the reasons for the different outcomes and it contends that the implementation of concentrated rural settlement practice under disaster-induced conditions is more effective than its introduction under development-driven conditions. The results of the analysis indicate that, in China, concentrated rural settlement is feasible in a context of post-disaster reconstruction. © 2014 The Author(s). Disasters © Overseas Development Institute, 2014.

  18. Estimating the concentration of aluminum-substituted hematite and goethite using diffuse reflectance spectrometry and rock magnetism: Feasibility and limitations

    NASA Astrophysics Data System (ADS)

    Hu, Pengxiang; Jiang, Zhaoxia; Liu, Qingsong; Heslop, David; Roberts, Andrew P.; Torrent, José; Barrón, Vidal

    2016-06-01

    Hematite and goethite in soils are often aluminum (Al) substituted, which can dramatically change their reflectance and magnetic properties and bias abundance estimates using diffuse reflectance spectroscopy (DRS) and magnetic techniques. In this study, synthetic Al-substituted hematites and goethites and two Chinese loess/paleosol sequences were investigated to test the feasibility and limitations of estimating Al-hematite and Al-goethite concentration. When Al substitution is limited (Al/(Al + Fe) molar ratio < ~8%), the reflectance spectrum provides a reliable estimate of the goethite/hematite concentration ratio. New empirical relationships between the DRS band intensity ratio and the true concentration goethite/hematite ratio are estimated as goethite/hematite = 1.56 × (I425 nm/I535 nm) or goethite/hematite = 6.32 × (I480 nm/I535 nm), where I425 nm, I480 nm, and I535 nm are the amplitudes of DRS second-derivative curves for characteristic bands at ~425 nm, ~480 nm, and ~535 nm, respectively. High Al substitution (> ~8%) reduces DRS band intensity, which leads to biased estimates of mineral concentration. Al substitution and grain size exert a control on coercivity distributions of hematite and goethite and, thus, affect the hard isothermal remanent magnetization. By integrating DRS and magnetic methods, we suggest a way to constrain hematite and goethite Al substitution in natural loess. Results indicate that hematite and goethite in Chinese loess have Al contents lower than ~8% and, thus, that DRS can be used to trace hematite and goethite concentration variations.

  19. Dynamic light scattering can determine platelet function

    NASA Astrophysics Data System (ADS)

    Lee, Nathan

    2011-10-01

    Platelet transfusions are life-saving procedures for patients who are bleeding or undergoing chemotherapy. The effectiveness of transfusions depends on the number of platelets transfused and the platelet function. Platelet function correlates with proportion of discoid to activated platelets, morphology response to temperature stress, and inversely correlates with microparticle content. ThromboLUX is a novel device that determines platelet function by measuring all of these characteristics using dynamic light scattering (DLS). During periods of stress, such as decreased temperature, cytoskeletal rearrangements will cause normal, discoid platelets to activate and become spiny spheres. The formation of pseudopods of various lengths facilitates the clotting cascade and also increases the apparent size of platelets. ThromboLUX uses a 37-20-37 C temperature cycle that mimics the bleeding, storage, and transfusion process. As the temperature fluctuates, DLS will measure the changing platelet hydrodynamic radius and the size of any microparticles present. ThromboLUX analysis of platelet concentrates in vitro would allow determination of high platelet function units before transfusion and would therefore improve transfusion outcomes and patient safety. This study examined how DLS is able to distinguish between discoid and activated platelets as well as measure the parameters that contribute to high platelet function.

  20. Feasibility of using a two-wavelength photometer to estimate the concentration of circulating near-infrared extinguishing nanoparticles.

    PubMed

    Michalak, Gregory J; Anderson, Heather A; O'Neal, D Patrick

    2010-02-01

    We demonstrate a photometer based on pulse oximeter technology designed to test the feasibility of using non-invasive optics to quantify in vivo circulation parameters of optically-active particles by measuring changes in optical extinction introduced by the particles in a murine animal model. A real-time estimate of relative concentration was produced by collecting log-scaled bandpass pulsatile and non-pulsatile intensity (760 nm or 940 nm) near the extinction peak of the employed gold nanoshells and mathematically subtracting the pre-injection intensity through the murine subject. The circulation half-lives in four mice were estimated between 3 and 43 minutes compared to direct optical measurement of 5 microL blood draws with UV/Vis spectrophotometry which demonstrated nanoparticle extinctions ranging from 0.246 to 7.408 optical density (OD). A linear model fit relating the two methods produced an R2 value of 0.75. The 1.795 OD negative bias (-4.98 x 10(9) nanoparticles/ml) between the two methods describes the 35.5% (or 12.0 minutes) average error of prediction of the half-life. This report demonstrates that the circulation parameters of optically-active particles employed at therapeutically-relevant concentrations can be monitored in real-time using non-invasive optical techniques and advises further refinement.

  1. Numerical simulation of platelet margination in microcirculation

    NASA Astrophysics Data System (ADS)

    Zhao, Hong; Shaqfeh, Eric

    2009-11-01

    The adhesion of platelets to vascular walls is the first step in clotting. This process critically depends on the preferential concentration of platelets near walls. The presence of red blood cells, which are the predominant blood constituents, is known to affect the steady state platelet concentration and the dynamic platelet margination, but the underlying mechanism is not well understood to-day. We use a direct numerical simulation to study the platelet margination process, with particular emphasis on the Stokesian hydrodynamic interactions among red cells, platelets, and vessel walls. Well-known mechanical models are used for the shearing and bending stiffness of red cell membranes, and the stiffer platelets are modeled as rigid discoids. A boundary integral formulation is used to solve the flow field, where the numerical solution procedure is accelerated by a parallel O(N N) smooth particle-mesh Ewald method. The effects of red cell hematocrit and deformability will be discussed.

  2. Simple tube centrifugation for processing platelet-rich plasma in the horse

    PubMed Central

    Fontenot, Robin L.; Sink, Carolyn A.; Werre, Stephen R.; Weinstein, Nicole M.; Dahlgren, Linda A.

    2012-01-01

    This study evaluated the quality and bacteriologic safety of platelet-rich plasma (PRP) produced by 3 simple, inexpensive tube centrifugation methods and a commercial system. Citrated equine blood collected from 26 normal horses was processed by 4 methods: blood collection tubes centrifuged at 1200 and 2000 × g, 50-mL conical tube, and a commercial system. White blood cell (WBC), red blood cell (RBC), and platelet counts and mean platelet volume (MPV) were determined for whole blood and PRP, and aerobic and anaerobic cultures were performed. Mean platelet concentrations ranged from 1.55- to 2.58-fold. The conical method yielded the most samples with platelet concentrations greater than 2.5-fold and within the clinically acceptable range of > 250 000 platelets/λL. White blood cell counts were lowest with the commercial system and unacceptably high with the blood collection tubes. The conical tube method may offer an economically feasible and comparatively safe alternative to commercial PRP production systems. PMID:23729823

  3. Hemolysis after ABO-incompatible platelet transfusions.

    PubMed

    Chow, M P; Yung, C H; Hu, H Y; Tzeng, C H

    1991-08-01

    An 18 year old girl, with acute myeloid leukemia, developed progressive hemolysis after receiving multiple transfusions with ABO-incompatible platelets. It was caused by passive transfusion of anti-A and -B isoagglutinin from the donor plasma. Her hemoglobin level returned to normal after giving group compatible or pooled and reduced volume platelet concentrates. Transfusing group-incompatible platelets is not contraindicated, but donor plasma reduction should be considered for those patients who need prolonged platelet support. Testing for isoagglutinin titer in group O donors is an alternate method to reduce the incidence of plasma-induced hemolysis in group-incompatible platelet transfusions.

  4. Effect of Sclerovit on endothelial dysfunction, hemorheological parameters, platelet aggregation, plasma concentration of homocysteine and progression of atherosclerosis in patients with vascular diseases.

    PubMed

    Horvath, Beata; Szapary, Laszlo; Debreceni, Laszlo; Feher, Gergely; Kenyeres, Peter; Fulop, Adrienn; Battyani, Istvan; Toth, Kalman

    2009-01-01

    In our prospective study the effect of Sclerovit (0.8 mg folic acid, 20 mug vitamin B12,5 mg vitamin B6,100 mg vitamin E) on inflammatory markers, hemorheological parameters, platelet aggregation, von Willebrand factor activity as a marker of endothelium dysfunction, plasma lipids, plasma levels of folic acid, vitamin B12 and homocysteine (hcy), flow mediated vasodilatation (FMD) and thickness of carotis intima-media after 1 and 6 months of treatment in patients with vascular diseases (10 patients took 1 capsule, 10 patients 2 capsules of Sclerovit and 10 patients placebo) was determined.Plasma level of vitamin B12, folic acid and elongation index of red blood cells (RBC) increased significantly (p<0.05-0.001), hcy and triglyceride concentrations decreased significantly (p<0.05-0.001) in patients taking Sclerovit. HDL-cholesterol, RBC count, hematocrit, plasma and whole blood viscosity increased significantly (p<0.05-0.001) both in patients taking placebo or vitamins. Fibrinogen and CRP showed a significant (p<0.05-0.01) increase in patients on placebo, but did not change in patients on Sclerovit therapy. FMD showed a significant (p<0.05) amelioration in patients on 1 capsule of Sclerovit.Beside the favorable effects of Sclerovit on some of the measured parameters, the observed deterioration in hemorheological parameters can correlate with the contradictory results of large prospective studies with vitamins.

  5. The Augmentation of a Collagen/Glycosaminoglycan Biphasic Osteochondral Scaffold with Platelet-Rich Plasma and Concentrated Bone Marrow Aspirate for Osteochondral Defect Repair in Sheep

    PubMed Central

    Henson, Frances; Skelton, Carrie; Herrera, Emilio; Brooks, Roger; Fortier, Lisa A.; Rushton, Neil

    2012-01-01

    Objective: This study investigates the combination of platelet-rich plasma (PRP) or concentrated bone marrow aspirate (CBMA) with a biphasic collagen/glycosaminoglycan (GAG) osteochondral scaffold for the treatment of osteochondral defects in sheep. Design: Acute osteochondral defects were created in the medial femoral condyle (MFC) and the lateral trochlea sulcus (LTS) of 24 sheep (n = 6). Defects were left empty or filled with a 6 × 6-mm scaffold, either on its own or in combination with PRP or CBMA. Outcome measures at 6 months included mechanical testing, International Cartilage Repair Society (ICRS) repair score, modified O’Driscoll histology score, qualitative histology, and immunohistochemistry for type I, II, and VI collagen. Results: No differences in mechanical properties, ICRS repair score, or modified O’Driscoll score were detected between the 4 groups. However, qualitative assessments of the histological architecture, Safranin O content, and collagen immunohistochemistry indicated that in the PRP/scaffold groups, there was a more hyaline cartilage–like tissue repair. In addition, the addition of CBMA and PRP to the scaffold reduced cyst formation in the subchondral bone of healed lesions. Conclusion: There was more hyaline cartilage–like tissue formed in the PRP/scaffold group and less subchondral cystic lesion formation in the CBMA and PRP/scaffold groups, although there were no quantitative differences in the repair tissue formed. PMID:26069645

  6. Degradation of methylene blue (MB) using ZnO/CeO2/nanographene platelets (NGP) photocatalyst: Effect of various concentration of NGP

    NASA Astrophysics Data System (ADS)

    Tju, H.; Shabrany, H.; Taufik, A.; Saleh, R.

    2017-07-01

    ZnO/CeO2 photocatalysts were loaded on to nanographene platelets (NGP) layer sheets to form ZnO/CeO2/NGP composites. The synthesized process was achieved by using two-step methods: Sol-gel followed by the hydrothermal method, the concentration of NGP was varied by 5, 10, and 15 wt.%. The as-prepared ZnO/CeO2/NGP samples were characterized by X-ray diffraction (XRD) and Brunner Emett-Teller (BET). The XRD spectra of all samples exhibit a good agreement with hexagonal wurtzite ZnO and face centered cubic phase of CeO2, additionally a new peak could be indexed for graphitic-like structures from NGP. The BET result shows that the incorporation of NGP could enhance the specific surface area of ZnO/CeO2 composites. Furthermore, it is found that addition of NGP in ZnO/CeO2 composites could enhance photocatalytic activities in methylene blue (MB) dye degradation compared to ZnO/CeO2 composites. Our results show that addition of 10 wt.% NGP to ZnO/CeO2 composites exhibits the highest photocatalytic activity. The enhancement of photocatalytic activities can be ascribed to the function of NGP as trap state for the electron. The scavenger tests results indicated that the photo-generated hole would play an important role in the degradation of MB.

  7. Platelet function and constituents of platelet rich plasma.

    PubMed

    Pelletier, M H; Malhotra, A; Brighton, T; Walsh, W R; Lindeman, R

    2013-01-01

    Platelet Rich Plasma (PRP) therapies require blood to be processed prior to application, however, the full assessment of the output of platelet sequestration devices is lacking. In this study the products of the Autologous Fluid Concentrator (Circle BiologicsTM, Minneapolis, MN) and the Gravitational Platelet Separation System (GPS, Biomet, Warsaw, IN, USA) were evaluated in terms of platelet viability and PRP constituents. The AFC and GPS produced 6.4 (±1.0) ml and 6.3 (±0.4) ml of PRP, with platelet recovery of 46.4% (±14.7%) and 59.8% (±24.2%) producing fold increases of platelets of 4.19 (±1.62) and 5.19 (±1.62), respectively. Fibrinogen concentration was increased above baseline PPP produced with the AFC. pH was lower for both of the processed samples than for whole blood. White Blood Cell count was increased around 5 fold. Functional tests showed preserved viability with both devices. This represents essential knowledge that every treating physician should have before they can confidently administer PRP therapy produced by any method. These are the first published results of platelet function for the GPS system and the first performance results of the AFC system. The PRP produced is classified according to broad classifications as Leukocyte-PRP (L-PRP) for both devices.

  8. Measurement of platelet aggregation, independently of patient platelet count: a flow-cytometric approach.

    PubMed

    Vinholt, P J; Frederiksen, H; Hvas, A-M; Sprogøe, U; Nielsen, C

    2017-06-01

    Essentials Platelet function may influence bleeding risk in thrombocytopenia, but useful tests are needed. A flow cytometric platelet aggregation test independent of the patient platelet count was made. Platelet aggregation was reduced in thrombocytopenic patients with hematological cancer. High platelet aggregation ruled out bleeding tendency in thrombocytopenic patients. Background Methods for testing platelet aggregation in thrombocytopenia are lacking. Objective To establish a flow-cytometric test of in vitro platelet aggregation independently of the patient's platelet count, and examine the association of aggregation with a bleeding history in thrombocytopenic patients. Patients/methods We established a flow-cytometric assay of platelet aggregation, and measured samples from healthy individuals preincubated with antiplatelet drugs, and samples from two patients with inherited platelet disorders. Then, we included 19 healthy individuals and 20 patients with platelet counts of ≤ 50 × 10(9) L(-1) , diagnosed with acute myeloid leukemia or myelodysplastic syndrome. We measured platelet aggregation and platelet activation by platelet surface expression of activated glycoprotein IIb-IIIa, P-selectin and CD63 after addition of agonists: collagen-related peptide, thrombin receptor-activating peptide (TRAP), and ADP. Results The platelet aggregation assay showed a low intraserial coefficient of variation of ≤ 3%. Similar results were obtained for platelet-rich plasma and isolated platelets at platelet counts of > 10 × 10(9) L(-1) ; otherwise, platelet isolation was required. The platelet aggregation percentage decreased with increasing antiplatelet drug concentration. Platelet aggregation in patients was reduced as compared with healthy individuals: 42% (interquartile range [IQR] 27-58) versus 66% (IQR 60-67) for TRAP; 41% (IQR 25-48) versus 70% (IQR 69-72) for collagen-related peptide; and 44% (IQR 30-53) versus 65% (IQR 46-72) for ADP. Platelet activation after

  9. Acetal phosphatidic acids: novel platelet aggregating agents.

    PubMed

    Brammer, J P; Maguire, M H; Walaszek, E J; Wiley, R A

    1983-05-01

    1 Palmitaldehyde, olealdehyde and linolealdehyde acetal phosphatidic acids induced rapid shape change and dose-dependent biphasic aggregation of human platelets in platelet-rich plasma; aggregation was reversible at low doses and irreversible at high doses of the acetal phosphatidic acids. The palmitaldehyde congener elicited monophasic dose-dependent aggregation of sheep platelets in platelet-rich plasma.2 The threshold concentration for palmitaldehyde acetal phosphatidic acid (PGAP)-induced platelet aggregation was 2.5-5 muM for human platelets and 0.25-0.5 muM for sheep platelets. PGAP was 4-5 times as potent versus human platelets as the olealdehyde and linolealdehyde acetal phosphatidic acids, which were equipotent.3 PGAP-induced irreversible aggregation of [(14)C]-5-hydroxytryptamine ([(14)C]-5-HT)-labelled human platelets in platelet-rich plasma was accompanied by release of 44.0+/-2.4% (s.e.) of the platelet [(14)C]-5-HT; reversible aggregation was not associated with release. In contrast, PGAP-induced release of [(14)C]-5-HT-labelled sheep platelets was dose-dependent.4 The adenosine diphosphate (ADP) antagonist, 2-methylthio-AMP, and the cyclo-oxygenase inhibitor, aspirin, abolished PGAP-induced second phase aggregation and release in human platelets but did not affect the first, reversible, phase of aggregation. Both the first and second phases of PGAP-induced aggregation were abolished by chlorpromazine, by the phospholipase A(2) inhibitor, mepacrine, and by nmolar concentrations of prostaglandin E(1) (PGE(1)); these agents abolished the second, but not the first phase of ADP-induced aggregation.5 The related phospholipids, lecithin, lysolecithin and phosphatidic acid, at <100 muM, neither induced aggregation of human platelets in platelet-rich plasma, nor modified PGAP-induced aggregation; 1-palmityl lysophosphatidic acid elicited aggregation of human platelets at a threshold concentration of 100 muM.6 It is concluded that the acetal phosphatidic acids

  10. Producing accurate platelet counts for platelet rich plasma: validation of a hematology analyzer and preparation techniques for counting.

    PubMed

    Woodell-May, Jennifer E; Ridderman, Dayna N; Swift, Matthew J; Higgins, Joel

    2005-09-01

    Platelet rich plasma (PRP) has been shown to clinically accelerate healing of both soft and hard tissues. As a result, it has gained increasing popularity. However, the clinical effectiveness of each type of PRP preparation method can vary in technique and efficiency, and current methods to evaluate the platelet concentration efficiency of PRP systems have several limitations. Therefore, the purpose of this study was to validate an automated hematology analyzer, the Cell-Dyn 3700, to accurately count platelets in concentration ranges of approximately 2,000,000-4,800,000 platelets/microL. PRP platelets were counted by way of a manual counting method and on the Cell-Dyn 3700, and the statistical evaluation indicated no difference between the groups (P > 0.05). Dilution of the PRP was not required, and accurate platelet counts could be achieved up to platelet concentrations of 4,800,000 platelets/microL. PRPs must be resuspended on a rocker for at least 5 minutes before platelet counts, and the entire PRP sample must be resuspended to allow for equal distribution of platelets before counting. With use of the validated Cell-Dyn 3700, a platelet concentrate system was used to prepare 153 PRPs. The baseline whole blood platelet concentration (328,000 platelets/microL +/- 69,000 platelets/microL) and the average PRP samples (2,645,000 platelets/microL +/- 680,000 platelets/microL) were compared, resulting in an eightfold increase in concentration and an average platelet percent recovery of approximately 76%. Automated hematology analyzers can be used to accurately count platelets in PRP given the system has been validated appropriately and the PRP samples are prepared properly to provide adequate platelet suspension.

  11. False-positive alarms for bacterial screening of platelet concentrates with BacT/ALERT new-generation plastic bottles: a multicenter pilot study.

    PubMed

    Hundhausen, T; Müller, T H

    2005-08-01

    The microbial detection system BacT/ALERT (bioMérieux) is widely used to monitor bacterial contamination of platelet concentrates (PCs). Recently, the manufacturer introduced polycarbonate culture bottles and a modified pH-sensitive liquid emulsion sensor as microbial growth indicator. This reconfigured assay was investigated in a routine setting. In each of eight transfusion centers, samples from 500 consecutive PCs were monitored for 1 week. For all PCs with a positive BacT/ALERT signal, retained samples and, if available, original PC containers and concomitant red blood cell concentrates were analyzed independently. Initially BacT/ALERT-positive PCs without bacterial identification in any sample were defined as false-positive. BacT/ALERT-positive PCs with bacteria in the first sample only were called potentially positive. PCs with bacteria in the first sample and the same strain in at least one additional sample were accepted as positive. Five PCs (0.13%) were positive, 9 PCs (0.23%) were potentially positive, and 35 PCs (0.9%) were false-positive. The rate of false-positive BacT/ALERT results varied substantially between centers (<0.2%-3.2%). Tracings from false-positive cultures lacked an exponential increase of the signal during incubation. Most of these false-positives were due to malfunctioning cells in various BacT/ALERT incubation units. Careful assessment of individual tracings of samples with positive signals helps to identify malfunctioning incubation units. Their early shutdown or replacement minimizes the high rate of unrectifiable product rejects attributed to false-positive alarms and avoids unnecessary concern of doctors and patients after conversion to a reconfigured BacT/ALERT assay.

  12. High glucose concentration induces the overexpression of transforming growth factor-beta through the activation of a platelet-derived growth factor loop in human mesangial cells.

    PubMed Central

    Di Paolo, S.; Gesualdo, L.; Ranieri, E.; Grandaliano, G.; Schena, F. P.

    1996-01-01

    High glucose concentration has been shown to induce the overexpression of transforming growth factor (TGF)-beta 1 mRNA and protein in different cell types, including murine mesangial cells, thus possibly accounting for the expansion of mesangial extracellular matrix observed in diabetic glomerulopathy. In the present study, we evaluated platelet-derived growth factor (PDGF) B-chain and PDGF-beta receptor gene expression in human mesangial cells (HMCs) exposed to different concentrations of glucose and then sought a possible relationship between a PDGF loop and the modulation of TGF-beta 1 expression. HMC [3H]thymidine incorporation was upregulated by 30 mmol/L glucose (HG) up to 24 hours, whereas it was significantly inhibited at later time points. Neutralizing antibodies to PDGF BB abolished the biphasic response to HG, whereas anti-TGF-beta antibodies reversed only the late inhibitory effect of hyperglycemic medium. HG induced an early and persistent increase of PDGF B-chain gene expression, as evaluated by reverse transcriptase polymerase chain reaction, whereas PDGF-beta receptor mRNA increased by twofold after 6 hours, thereafter declining at levels 70% lower than in controls after 24 hours. 125I-Labeled PDGF BB binding studies in HMCs exposed to HG for 24 hours confirmed the decrease of PDGF-beta receptor expression. TGF-beta 1-specific transcripts showed 43 and 78% increases after 24 and 48 hours of incubation in HG, respectively, which was markedly diminished by anti-PDGF BB neutralizing antibodies or suramin. We conclude that HG induces an early activation of a PDGF loop that, in turn, causes an increase of TGF-beta 1 gene expression, thus modulating both HMC proliferation and mesangial matrix production. Images Figure 2 Figure 3 Figure 4 Figure 5 PMID:8952542

  13. Development of a method to quantify platelet adhesion and aggregation under static conditions

    PubMed Central

    Baker-Groberg, Sandra M.; Cianchetti, Flor A.; Phillips, Kevin G.; McCarty, Owen J.T.

    2014-01-01

    Platelets are important players in hemostasis and thrombosis. Thus, accurate assessment of platelet function is crucial for identifying platelet function disorders and measuring the efficacy of antiplatelet therapies. We have developed a novel platelet aggregation technique that utilizes the physical parameter of platelet concentration in conjunction with volume and mass measurements to evaluate platelet adhesion and aggregation. Platelet aggregates were formed by incubating purified platelets on fibrinogen- or fibrillar collagen-coated surfaces at platelet concentrations ranging from 20,000 to 500,000 platelets/ L. Platelets formed aggregates under static conditions in a platelet concentration-dependent manner, with significantly greater mean volume and mass at higher platelet concentrations ( 400,000 platelets/ L). We show that a platelet glycoprotein IIb/IIIa inhibitor abrogated platelet-platelet aggregation, which significantly reduced the volume and mass of the platelets on the collagen surface. This static platelet aggregation technique is amenable to standardization and represents a useful tool to investigate the mechanism of platelet activation and aggregation under static conditions. PMID:24883127

  14. Development of a method to quantify platelet adhesion and aggregation under static conditions.

    PubMed

    Baker-Groberg, Sandra M; Cianchetti, Flor A; Phillips, Kevin G; McCarty, Owen J T

    2014-06-01

    Platelets are important players in hemostasis and thrombosis. Thus, accurate assessment of platelet function is crucial for identifying platelet function disorders and measuring the efficacy of antiplatelet therapies. We have developed a novel platelet aggregation technique that utilizes the physical parameter of platelet concentration in conjunction with volume and mass measurements to evaluate platelet adhesion and aggregation. Platelet aggregates were formed by incubating purified platelets on fibrinogen- or fibrillar collagen-coated surfaces at platelet concentrations ranging from 20,000 to 500,000 platelets/ L. Platelets formed aggregates under static conditions in a platelet concentration-dependent manner, with significantly greater mean volume and mass at higher platelet concentrations ( 400,000 platelets/ L). We show that a platelet glycoprotein IIb/IIIa inhibitor abrogated platelet-platelet aggregation, which significantly reduced the volume and mass of the platelets on the collagen surface. This static platelet aggregation technique is amenable to standardization and represents a useful tool to investigate the mechanism of platelet activation and aggregation under static conditions.

  15. A dynamic magnetic shift method to increase nanoparticle concentration in cancer metastases: a feasibility study using simulations on autopsy specimens

    PubMed Central

    Nacev, Alek; Kim, Skye H; Rodriguez-Canales, Jaime; Tangrea, Michael A; Shapiro, Benjamin; Emmert-Buck, Michael R

    2011-01-01

    A nanoparticle delivery system termed dynamic magnetic shift (DMS) has the potential to more effectively treat metastatic cancer by equilibrating therapeutic magnetic nanoparticles throughout tumors. To evaluate the feasibility of DMS, histological liver sections from autopsy cases of women who died from breast neoplasms were studied to measure vessel number, size, and spatial distribution in both metastatic tumors and normal tissue. Consistent with prior studies, normal tissue had a higher vascular density with a vessel-to-nuclei ratio of 0.48 ± 0.14 (n = 1000), whereas tumor tissue had a ratio of 0.13 ± 0.07 (n = 1000). For tumors, distances from cells to their nearest blood vessel were larger (average 43.8 μm, maximum 287 μm, n ≈ 5500) than normal cells (average 5.3 μm, maximum 67.8 μm, n ≈ 5500), implying that systemically delivered nanoparticles diffusing from vessels into surrounding tissue would preferentially dose healthy instead of cancerous cells. Numerical simulations of magnetically driven particle transport based on the autopsy data indicate that DMS would correct the problem by increasing nanoparticle levels in hypovascular regions of metastases to that of normal tissue, elevating the time-averaged concentration delivered to the tumor for magnetic actuation versus diffusion alone by 1.86-fold, and increasing the maximum concentration over time by 1.89-fold. Thus, DMS may prove useful in facilitating therapeutic nanoparticles to reach poorly vascularized regions of metastatic tumors that are not accessed by diffusion alone. PMID:22131836

  16. The platelet defect in leukemia. Platelet ultrastructure, adenine nucleotide metabolism, and the release reaction.

    PubMed Central

    Cowan, D H; Graham, R C; Baunach, D

    1975-01-01

    The ultrastructure and adenine nucleotide metabolism of platelets from patients with acute leukemia were studied to elucidate possible mechanisms for the platelet dysfunction observed in this clinical setting. Nonstimulated (resting) platelets from leukemic patients varied greatly in size; exhibited marked variation in the number of alpha granules present per cell; had poorly delineated circumferential bands of microtubules; and often grossly dilated open channel systems or cytoplasmic vacuolization. The intracellular concentrations of ATP and ADP were significantly below normal, and the specific radioactivity of ATP and ADP of nonstimulated platelets in leukemia was equivalent to or exceeded that seen in stimulated normal platelets. Addition of ADP or collagen to platelets from leukemic patients was followed by retarded and incomplete shape change, delayed and incomplete centripetal migration of subcellular organelles, impaired degranulation, and the formation of loose aggregates composed of relatively few platelets. Stimulation of "leukemic" platelets with collagen led to the release of significantly subnormal amounts of ATP and ADP and no significant change in the specific radioactivity of the intracellular nucleotides. In contrast to the results in normal platelets, the conversion of ATP to inosine monophosphate and hypoxanthine in platelets in leukemia failed to increase significantly with collagen stimulation. The results indicate that abnormalities exist in the storage pool of adenine nucleotides and the release mechanism of platelets in acute leukemia. These defects appear to contribute to an impairment in the release reaction in these platelets. Many of the ultrastructural and metabolic defects seen in acute leukemia occur in platelets in preleukemia. Images PMID:45818

  17. [Assessment study on a set of platelet-rich plasma preparation].

    PubMed

    Li, Ming; Zhang, Changqing; Yuan, Ting; Chen, Shengbao; Lü, Ruju

    2011-01-01

    To calculate the recovery rate and enrichment factor and to analyse the correlation by measuring the concentrations of platelets, leukocyte, and growth factors in platelet-rich plasma (PRP) so as to evaluate the feasibility and stability of a set of PRP preparation. The peripheral blood (40 mL) was collected from 30 volunteers accorded with the inclusion criteria, and then 4 mL PRP was prepared using the package produced by Shandong Weigao Group Medical Polymer Company Limited. Automatic hematology analyzer was used to count the concentrations of platelets and leukocyte in whole blood and PRP. The enrichment factor and recovery rate of platelets or leukocyte were calculated; the platelet and leukocyte concentrations of male and female volunteers were measured, respectively. The concentrations of platelet-derived growth factor (PDGF), transforming growth factor beta (TGF-beta), and vascular endothelial growth factor (VEGF) were assayed by ELISA. The platelet concentrations of whole blood and PRP were (131.40 +/- 29.44) x 10(9)/L and (819.47 +/- 136.32) x 10(9)/L, respectively, showing significant difference (t = 27.020, P = 0.000). The recovery rate of platelets was 60.85% +/- 8.97%, and the enrichment factor was 6.40 +/- 1.06. The leukocyte concentrations of whole blood and PRP were (5.57 +/- 1.91) x 10(12)/L and (32.20 +/- 10.42) x 10(12)/L, respectively, showing significant difference (t = 13.780, P = 0.000). The recovery rate of leukocyte was 58.30% +/- 19.24%, and the enrichment factor was 6.10 +/- 1.93. The concentrations of platelets and leukocyte in PRP were positively correlated with the platelet concentration (r = 0.652, P = 0.000) and leukocyte concentration (r = 0.460, P = 0.011) in whole blood. The concentrations of platelet and leukocyte in PRP between male and female were not significantly different (P > 0.05). The concentrations of PDGF, TGF-beta, and VEGF in PRP were (698.15 +/- 64.48), (681.36 +/- 65.90), and (1071.55 +/- 106.04) ng/mL, which were

  18. Insulin enhances platelet activation in vitro.

    PubMed

    Yngen, M; Li, N; Hjemdahl, P; Wallén, N H

    2001-10-15

    Diabetes mellitus is associated with an increased risk of atherothrombotic complications. There is accumulating evidence of platelet hyperreactivity in diabetes, which may be of importance in the pathogenesis of diabetic vascular complications. Platelets possess insulin receptors, but their physiological relevance is not clear, and data on insulin effects on platelet function in the literature are less than consistent. We therefore investigated the influence of insulin on platelet activation in vitro. Fasting blood samples were collected in 20 healthy males, using citrate or hirudin as anticoagulants. Platelet activation was measured as platelet P-selectin expression and fibrinogen binding using whole blood flow cytometry in unstimulated and adenosine diphosphate (ADP) stimulated samples, incubated with 0-10000 microU/ml insulin for 20 min. The effect of insulin (30 or 300 microU/ml, incubated for 3 min) on platelet aggregation was studied using Born aggregometry in platelet-rich plasma (PRP). Insulin enhanced platelet fibrinogen binding more than P-selectin expression in unstimulated and ADP stimulated samples (P<.001 by analysis of variance [ANOVA]; n=20). Insulin (30 or 300 microU/ml) also enhanced ADP-induced platelet aggregation in PRP (P<.01 or P<.001; n=14). The platelet activating effect of insulin was verified in hirudinized samples (n=12), indicating that it was not dependent on unphysiologically low extracellular calcium concentrations. Thus, insulin enhances platelet activation in vitro, independently of extracellular calcium concentrations. Beneficial effects of insulin treatment on platelet function in vivo are probably related to improved metabolic control, rather than to direct platelet stabilizing effects.

  19. Establishing proof of concept: Platelet-rich plasma and bone marrow aspirate concentrate may improve cartilage repair following surgical treatment for osteochondral lesions of the talus.

    PubMed

    Smyth, Niall A; Murawski, Christopher D; Haleem, Amgad M; Hannon, Charles P; Savage-Elliott, Ian; Kennedy, John G

    2012-07-18

    Osteochondral lesions of the talus are common injuries in the athletic patient. They present a challenging clinical problem as cartilage has a poor potential for healing. Current surgical treatments consist of reparative (microfracture) or replacement (autologous osteochondral graft) strategies and demonstrate good clinical outcomes at the short and medium term follow-up. Radiological findings and second-look arthroscopy however, indicate possible poor cartilage repair with evidence of fibrous infill and fissuring of the regenerative tissue following microfracture. Longer-term follow-up echoes these findings as it demonstrates a decline in clinical outcome. The nature of the cartilage repair that occurs for an osteochondral graft to become integrated with the native surround tissue is also of concern. Studies have shown evidence of poor cartilage integration, with chondrocyte death at the periphery of the graft, possibly causing cyst formation due to synovial fluid ingress. Biological adjuncts, in the form of platelet-rich plasma (PRP) and bone marrow aspirate concentrate (BMAC), have been investigated with regard to their potential in improving cartilage repair in both in vitro and in vitro settings. The in vitro literature indicates that these biological adjuncts may increase chondrocyte proliferation as well as synthetic capability, while limiting the catabolic effects of an inflammatory joint environment. These findings have been extrapolated to in vitro animal models, with results showing that both PRP and BMAC improve cartilage repair. The basic science literature therefore establishes the proof of concept that biological adjuncts may improve cartilage repair when used in conjunction with reparative and replacement treatment strategies for osteochondral lesions of the talus.

  20. Inhibition of activated protein C by platelets.

    PubMed Central

    Jane, S M; Mitchell, C A; Hau, L; Salem, H H

    1989-01-01

    Activated protein C (APC), an anticoagulant that acts by inactivating Factors Va and VIIIa, is dependent on a suitable surface for its action. In this study we examined the ability of human platelets to provide this surface and support APC-mediated anticoagulant effects. The activity of APC was examined in three systems: the Factor Xa recalcification time of Al(OH)3 adsorbed plasma, studies of thrombin generation in recalcified plasma, and assessment of the rate of inactivation of purified Factor Va. In comparison with phospholipid, intact platelets required significantly greater concentrations of APC to achieve a similar degree of anticoagulation. When washed platelet membranes were substituted for intact platelets, adequate support of APC was observed and the anticoagulant effect was similar to that obtained with phospholipid. Platelet releasate obtained by stimulation of platelets with thrombin and epinephrine contained an inhibitor that interfered with the ability of phospholipid and washed platelet membranes to catalyze the anticoagulant effects of APC. A noncompetitive inhibition was suggested by Dixon plot analysis of the interaction between platelet releasate and APC. The activity of the platelet APC inhibitor was immediate and was not enhanced by heparin, distinguishing it from the circulating protein C inhibitor. The presence of this inhibitor in the platelet and its release with platelet stimulation emphasizes the procoagulant role of this cell. PMID:2910909

  1. Agonist concentration-dependent differential responsivity of a human platelet purinergic receptor: pharmacological and kinetic studies of aggregation, deaggregation and shape change responses mediated by the purinergic P2Y1 receptor in vitro.

    PubMed

    Maayani, Saul; Schwarz, Todd E; Patel, Nayana D; Craddock-Royal, Barbara D; Tagliente, Thomas M

    2003-01-01

    Platelet shape change (SC), aggregation and deaggregation responses are integral components of hemostasis that are elicited and modulated in vivo by the simultaneous activation of several membrane receptors. Selective activation of the purinergic P2Y1 receptor in vivo elicits a sustained SC and a small, transient aggregation response that is reversed rapidly by a robust deaggregation response (Platelets 2003; 14: 89). Using a kinetics-based turbidimetric approach to study the modulation of these concurrent components of human platelet responses, we demonstrate that these P2Y1 receptor-related responses and a number of their kinetic and steady-state characteristics are differentially elicited and modulated. P2Y1 receptor agonist concentrations that elicited aggregation (pEC50 for ADP, 2-MeSADP; 5.88, 6.69) were 10-fold greater than those that elicited SC (7.33, 7.67). The magnitude of the aggregation response was agonist concentration-dependent, saturable and was associated with an agonist concentration-dependent deceleration of the deaggregation response. Gi-coupled receptor (alpha 2A-adrenoceptor, EP3 and P2Y12 receptors) agonists also enhanced aggregation through deceleration of the deaggregation response, and an inhibitor of PI3K activity (wortmannin) inhibited aggregation through acceleration of the deaggregation response. Neither treatment affected the extent or the kinetics of the SC response. The aggregation but not the SC response was rapidly desensitized by P2Y1 receptor activation by ADP. The affinity of the presence of a single P2Y1 receptor subtype. The differential characteristics and modulation of the SC and aggregation responses by a single receptor support the idea that different signaling pathways activated at different occupancy states of the same receptor underlie the two responses. P2Y1 receptor-mediated platelet aggregation and SC responses provide a convenient model for studying the phenomenon of agonist-directed signaling by differential

  2. Coronary CT angiography using low concentrated contrast media injected with high flow rates: Feasible in clinical practice.

    PubMed

    Mihl, Casper; Kok, Madeleine; Wildberger, Joachim E; Altintas, Sibel; Labus, David; Nijssen, Estelle C; Hendriks, Babs M F; Kietselaer, Bas L J H; Das, Marco

    2015-11-01

    Aim of this study was to test the hypothesis that peak injection pressures and image quality using low concentrated contrast media (CM) (240 mg/mL) injected with high flow rates will be comparable to a standard injection protocol (CM: 300 mg/mL) in coronary computed tomographic angiography (CCTA). One hundred consecutive patients were scanned on a 2nd generation dual-source CT scanner. Group 1 (n=50) received prewarmed Iopromide 240 mg/mL at an injection rate of 9 mL/s, followed by a saline chaser. Group 2 (n=50) received the standard injection protocol: prewarmed Iopromide 300 mg/mL; flow rate: 7.2 mL/s. For both protocols, the iodine delivery rate (IDR, 2.16 gI/s) and the total iodine load (22.5 gI) were kept identical. Injection pressure (psi) was continuously monitored by a data acquisition program. Contrast enhancement was measured in the thoracic aorta and all proximal and distal coronary segments. Subjective and objective image quality was evaluated between both groups. No significant differences in peak injection pressures were found between both CM groups (121 ± 5.6 psi vs. 120 ± 5.3 psi, p=0.54). Flow rates of 9 mL/s were safely injected without any complications. No significant differences in contrast-to-noise ratio, signal-to-noise ratio and subjective image quality were found (all p>0.05). No significant differences in attenuation levels were found in the thoracic aorta and all segments of the coronary arteries (all p>0.05). Usage of low iodine concentration CM and injection with high flow rates is feasible. High flow rates (9 mL/s) of Iopromide 240 were safely injected without complications and should not be considered a drawback in clinical practice. No significant differences in peak pressure and image quality were found. This creates a doorway towards applicability of a broad variety in flow rates and IDRs and subsequently more individually tailored injection protocols. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  3. In vitro platelet adhesion to dialysis membranes.

    PubMed

    Remuzzi, A; Boccardo, P; Benigni, A

    1991-01-01

    This work describes an in vitro system developed to quantitate platelet deposition on different dialysis membranes. The system is based on the use of small dialysis filters and reproduces the haemodynamic pattern of blood flowing through hollow fibres during in vivo dialysis. We have determined the in vitro platelet adhesion to cuprophan and to a non-cellulosic membrane, polymethylmethacrylate. When albumin concentration in the platelet suspension was low (0.35%) platelet deposition to cuprophan and to polymethylmethacrylate was comparable. When albumin concentration was increased to a physiological value (3.5%) platelet adhesion to both cuprophan and to polymethylmethacrylate membranes significantly decreased. This effect of albumin was greatest for the high-permeable polymethylmethacrylate membrane (BK). These data suggest that platelet membrane interaction is significantly influenced by circulating albumin.

  4. Development of white blood cell fragments, during the preparation and storage of platelet concentrates, as measured by using real-time polymerase chain reaction.

    PubMed

    Dijkstra-Tiekstra, M J; van der Schoot, C E; Pietersz, R N I; Huijgens, P C; van der Meer, P F; Reesink, H W

    2004-11-01

    White blood cell (WBC) fragments may cause human leucocyte antigen (HLA) immunization in recipients. We investigated the occurrence and production of WBC fragments in platelet concentrates (PCs) and plasma units, during storage and filtration, by using real-time polymerase chain reaction (PCR) and flow cytometry. To study the occurrence of WBC fragments, 'male' WBCs were spiked into double-filtered 'female' PCs in a concentration series of 0.03-100 WBCs/microl (n = 4 per level). To study the production of WBC fragments, 'male' WBCs were spiked into 'female' plasma units to 4 x 10(9) WBCs/l and stored at room temperature prior to filtration (n = 4 per storage time; t = 0, 24 or 48 h). DNA was measured by both albumin real-time PCR and Y real-time PCR. Intact WBCs were counted by using flow cytometry. The number of WBC fragments was calculated by subtracting cell-free DNA (real-time PCR on supernatant) and intact WBCs (flow cytometry) from the total DNA amount (real-time PCR). Spiking of 'male' WBCs into 'female' PCs showed that the Y real-time PCR is linear and has a reproducible quantitative range down to 0.03 WBC/microl, but that the albumin-PCR, in unspiked samples, revealed a total of 6-10 WBC equivalents/microl (eq/microl). After centrifugation, half of this was observed as cell-free DNA in the supernatant, suggesting that the remaining DNA is derived from WBC fragments. The number of intact WBCs, amount of cell-free DNA and number of WBC fragments after filtration increased significantly when filtration was delayed for up to 48 h, from 0.1 WBC/microl, 1.3 WBC eq/microl and 0.6 WBC eq/microl at t = 0 h to 25 WBC/microl, 38 WBC eq/microl and 57 WBC eq/microl at t = 48 h, respectively. WBC fragments occur in WBC-reduced PCs and increase when products are stored, prior to filtration, up to levels that are equivalent to the amounts of intact WBCs that induce HLA immunization (i.e. > 5 x 10(6)/unit).

  5. Platelet Function in Basset Hound Hereditary Thrombopathy.

    DTIC Science & Technology

    1986-01-01

    Chrono -Lume. Thromb Res 32(5):509, 1983. Mills DCB. Platelet aggregation and the adenylate cyclase system. In Platelets and Thromosis, DCB Mills and FI ...44 Figure ATP release to varying ADP concentrations ............. 9 Chrono -Lume potentiation of aggregation...............l0 Platelet ATP release...hours of blood collection. Storage pool adenine nucleotide release was monitored using the Lumi- Aggregometer ( Chrono -Log Corp., Havertown, Pa

  6. Platelet serotonin modulates immune functions.

    PubMed

    Mauler, M; Bode, C; Duerschmied, D

    2016-01-01

    This short review addresses immune functions of platelet serotonin. Platelets transport serotonin at a high concentration in dense granules and release it upon activation. Besides haemostatic, vasotonic and developmental modulation, serotonin also influences a variety of immune functions (mediated by different serotonin receptors). First, platelet serotonergic effects are directed against invading pathogens via activation and proliferation of lymphocytes, modulation of cytokine release, and recruitment of neutrophils to sites of acute inflammation by induction of selectin expression on endothelial cells. Second, serotonin levels are elevated in autoimmune diseases, such as asthma or rheumatoid arthritis, and during tissue regeneration after ischemia of myocardium or brain. Specific antagonism of serotonin receptors appears to improve survival after myocardial infarction or sepsis and to attenuate asthmatic attacks in animal models. It will be of great clinical relevance if these findings can be translated into human applications. In conclusion, targeting immune modulatory effects of platelet serotonin may provide novel therapeutic options for common health problems.

  7. [Platelet-washing solution optimization].

    PubMed

    Grossin, E; Chamfly, V

    2005-10-01

    Different washing and homogénéisation solutions are hereby analysed by comparing the evolution of functional indicators during the preservation of washed aphaeresis platelet concentrates: physiological pH 4.5 and 6 solutions, buffered physiological pH 6.8 glucose solution, and two physiological pH 7 citrate solutions with acetate. Prior acidification of platelet concentrates proved to be essential. Two washings with manual or automated technique, guarantee residual proteins at a level of less than 0.5 g. Solutions T-Sol Baxter or SSP Macopharma allow us to obtain a product that meet the PSL specifications. Routine since June 2004, washings are done with a physiological pH 6 solution, then homogeneised with T-Sol solution. Platelet recovery, swirling phenomenon, lack of agrgegates, pH maintenance, low increase in the platelet average volume and maintenance of intra-cell potassium level, suggest that platelet entirety is preserved beyond the product's expiration date. The platelet transfusion yield of these products is satisfactory.

  8. Platelet associated antibodies

    MedlinePlus

    ... medlineplus.gov/ency/article/003552.htm Platelet-associated antibodies blood test To use the sharing features on ... JavaScript. This blood test shows if you have antibodies against platelets in your blood. Platelets are a ...

  9. Thrombospondin-induced adhesion of human platelets.

    PubMed Central

    Tuszynski, G P; Kowalska, M A

    1991-01-01

    Washed human unactivated platelets attached and spread on thrombospondin (TSP)-coated microtiter plates. Platelet adhesion was promoted by divalent cations Mn2+, Mg2+, and Ca2+ as compared to buffer having all divalent cations complexed with EDTA. TSP-dependent adhesion was inhibited by anti-TSP fab fragments, an anti-TSP monoclonal antibody, an RGD-containing peptide, complex-specific anti-glycoprotein (GP)IIb-IIIa monoclonal antibodies (A2A9 or AP-2) and anti-VLA-2 monoclonal antibodies (6F1 and Gi9), but not by rabbit preimmune fab fragments, mouse IgG, an anti-GPIIIa monoclonal antibody, or monoclonal antibodies against either the human vitronectin receptor, glycocalicin, or GPIV. At saturating concentrations, anti-GPIIb-IIIa inhibited adhesion by 40-60%. Glanzman's thrombasthenic platelets, which lack GPIIb-IIIa, adhered to TSP to the same extent as anti-GPIIb-IIIa-treated normal platelets or 40-60% as well as untreated normal platelets. Antibody 6F1 (5-10 micrograms/ml) inhibited platelet adhesion of both normal and thrombasthenic platelets by 84-100%. Both VLA-2 antibodies also inhibited collagen-induced platelet adhesion, but had no effect on fibronectin-induced adhesion of normal platelets. These data indicate that platelets specifically adhere to TSP and that this adhesion is mediated through GPIIb-IIIa and/or VLA-2. Images PMID:2010551

  10. Bulk fluid phase behaviour of colloidal platelet-sphere and platelet-polymer mixtures.

    PubMed

    de las Heras, Daniel; Schmidt, Matthias

    2013-04-13

    Using a geometry-based fundamental measure density functional theory, we calculate bulk fluid phase diagrams of colloidal mixtures of vanishingly thin hard circular platelets and hard spheres. We find isotropic-nematic phase separation, with strong broadening of the biphasic region, upon increasing the pressure. In mixtures with large size ratio of platelet and sphere diameters, there is also demixing between two nematic phases with differing platelet concentrations. We formulate a fundamental measure density functional for mixtures of colloidal platelets and freely overlapping spheres, which represent ideal polymers, and use it to obtain phase diagrams. We find that, for low platelet-polymer size ratio, in addition to isotropic-nematic and nematic-nematic phase coexistence, platelet-polymer mixtures also display isotropic-isotropic demixing. By contrast, we do not find isotropic-isotropic demixing in hard-core platelet-sphere mixtures for the size ratios considered.

  11. Status Report on Cryopreservation of Human Platelets.

    DTIC Science & Technology

    1979-03-27

    anticoagulated with 63 mlPost-Thaw Platelet Count X Volume of Thawed Platelets of CPD. The blood was centrifuged+ * at 4500 g for 2.5 minutes at 22 ± 2...C to isolate PRP , the PRP was centrifuged51-Chromium Platelet Survival In Vivo and then resuspended in 0.5 ml at 4500 g to concentrate the platelets...the fol- anticoagulated with 2.5 ml of 1.2% manually. The esuspended plateletminor modifications: EDTA in saline, were obtained I manate w suseled

  12. The effects of drugs used in anaesthesia on platelet membrane receptors and on platelet function.

    PubMed

    Kozek-Langenecker, Sibylle A

    2002-06-01

    Platelet dysfunctions are known origins of perioperative bleeding disorders which are a major concern in the management of surgical patients. Among multiple factors, interactions of drugs used in anaesthesia with platelets have been implicated to aggravate the risk of haemorrhagic complications. This paper reviews in vitro and in vivo studies which have examined the effects of inhalational, intravenous, and local anaesthetics, opioids, and muscle relaxants on platelets. A brief summary of platelet physiology, function tests, and flow cytometric assessment of membrane receptors is included. Although the results of many studies have been conflicting, it appears that halothane, sevoflurane, and propofol inhibit platelet function in a reversible and dose-related manner at concentrations used clinically. Ilalothane affects the intracellular activating second messenger inositol triphosphate, platelet calcium homeostasis, thromboxane A2 formation, and the inhibiting signal transduction pathway including cyclic adenosine monophosphate. The proposed platelet inhibiting mechanism of sevoflurane involves the suppression of thromboxane A2 formation. Propofol appears to cause platelet dysfunctions by inhibiting calcium mobilisation upon agonist stimulation. Nitrous oxide causes a modest suppression of calcium mobilisation. An interaction of local anaesthetics with components in the platelet membrane appears to account for their inhibiting effect, but only at concentrations far higher than that found during clinical use. A clinically relevant antithrombotic effect of regional anaesthesia has been observed, though. Isoflurane, enflurane, desflurane, barbiturates, etomidate, opioids, and muscle relaxants seem to have negligible effects on platelets at therapeutic concentrations. Anaesthetists should be aware of the potential impairment of the coagulation profile by anaesthetic agents.

  13. Blood platelet aggregation and personality traits.

    PubMed

    Jenkins, C D; Thomas, G; Olewine, D; Zyzanski, S J; Simpson, M T; Hames, C G

    1975-12-01

    Changes in blood platelet aggregation may precipitate episodes of arterial occlusive diseases. Little is known, however, regarding the influence of psychological traits, emotional states and other behavioral stressors on platelet aggregation phenomena. This study examined 46 healthy college men at rest and after submaximal treadmill exercise. Associations were found between the duration of platelet aggregation and a number of scores from the California Psychological Inventory and self-administered anxiety scales. The more socially adequate, poised and dominant persons--those with more mature ego development and less overt anxiety--had platelets with more prolonged aggregation reactions to the in vitro introduction of noradrenalin. Irreversible aggregation of platelets occurred more regularly to lower in vitro concentrations of noradrenalin in platelet samples drawn from subjects who were less anxious and tended to be more rigidly defensive. It is premature to attempt to derive clinical implications from this exploratory work, but some implications for the design of future research are discussed.

  14. Identification of platelet refractoriness in oncohematologic patients

    PubMed Central

    Ferreira, Aline Aparecida; Zulli, Roberto; Soares, Sheila; de Castro, Vagner; Moraes-Souza, Helio

    2011-01-01

    OBJECTIVES: To identify the occurrence and the causes of platelet refractoriness in oncohematologic patients. INTRODUCTION: Platelet refractoriness (unsatisfactory post-transfusion platelet increment) is a severe problem that impairs the treatment of oncohematologic patients and is not routinely investigated in most Brazilian services. METHODS: Forty-four episodes of platelet concentrate transfusion were evaluated in 16 patients according to the following parameters: corrected count increment, clinical conditions and detection of anti-platelet antibodies by the platelet immunofluorescence test (PIFT) and panel reactive antibodies against human leukocyte antigen class I (PRA-HLA). RESULTS: Of the 16 patients evaluated (median age: 53 years), nine (56%) were women, seven of them with a history of pregnancy. An unsatisfactory increment was observed in 43% of the transfusion events, being more frequent in transfusions of random platelet concentrates (54%). Platelet refractoriness was confirmed in three patients (19%), who presented immunologic and non-immunologic causes. Alloantibodies were identified in eight patients (50%) by the PIFT and in three (19%) by the PRA-HLA. Among alloimmunized patients, nine (64%) had a history of transfusion, and three as a result of pregnancy (43%). Of the former, two were refractory (29%). No significant differences were observed, probably as a result of the small sample size. CONCLUSION: The high rate of unsatisfactory platelet increment, refractoriness and alloimmunization observed support the need to set up protocols for the investigation of this complication in all chronically transfused patients, a fundamental requirement for the guarantee of adequate management. PMID:21437433

  15. Short-term exposure of platelets to glucose impairs inhibition of platelet aggregation by cyclooxygenase inhibitors.

    PubMed

    Kobzar, Gennadi; Mardla, Vilja; Samel, Nigulas

    2011-01-01

    Aspirin treatment reduces cardiovascular events and deaths in high-risk non-diabetic patients, but not in patients suffering from diabetes. In these patients, hyperglycemia has been found to cause reduced platelet sensitivity to aspirin. It is supposed that long-term exposure of platelets to glucose leads to non-enzymatic glycosylation and impairs aspirin inhibition of platelet aggregation. On the other hand, short-term exposure of platelets to glucose also attenuates the effect of aspirin on platelets. The aim of the present work was to analyse the effect of short-term exposure of glucose on the inhibition of platelet aggregation by aspirin and other cyclooxygenase (COX) inhibitors. Already a 15 min exposure of platelets to glucose impaired aspirin inhibition of the platelet aggregation induced by collagen, thrombin, adenosine diphosphate (ADP), and arachidonic acid (AA). Aspirin inhibition of platelet aggregation in platelet-rich plasma (PRP) was attenuated by 5.6, 11.2, 16.8, and 22.4 mM of glucose in a concentration-dependent way. The same effect was observed with indomethacin and acetaminophen used as cyclooxygenase inhibitors instead of aspirin. N-methyl-L-arginine, an inhibitor of nitric oxide synthase, prevented the effect of glucose on aspirin, indomethacin and acetaminophen inhibition of platelet aggregation. Other monosaccharides, for example fructose and galactose, impaired aspirin inhibition as did glucose. Lactic acid (0.1, 0.2, 0.4, 0.8 mM), the end product of anaerobic glycolysis in platelets, impaired the inhibition of platelet aggregation with aspirin in a concentration-dependent way but did not affect indomethacin. It is suggested that lactic acid might be a mediator of the effect of glucose on aspirin inhibition in platelets.

  16. A randomised controlled feasibility study investigating the use of eccentric and concentric strengthening exercises in the treatment of rotator cuff tendinopathy.

    PubMed

    Bateman, Marcus; Adams, Nicola

    2014-01-01

    To conduct a feasibility study to compare concentric and eccentric rotator cuff strengthening exercises for rotator cuff tendinopathy. A total of 11 patients with rotator cuff tendinopathy who were on the waiting list for arthroscopic subacromial decompression surgery were randomised to perform eccentric rotator cuff strengthening exercises, concentric strengthening exercises or no exercises. Patients were evaluated in terms of levels of pain and function using the Oxford Shoulder Score and a Visual Analogue Scale initially, at 4 weeks and at 8 weeks. The study design was found to be acceptable to patients and achieved a high level of 86% compliance. The drop-out rate was 0%. Two patients performing eccentric strengthening exercises improved sufficiently to cancel their planned surgery. Further research in this area is recommended. The study design was feasible and power calculations have been conducted to aid future research planning.

  17. A manual method to obtain platelet rich plasma.

    PubMed

    Marques, Fabiana Paulino; Ingham, Sheila Jean McNeill; Forgas, Andrea; Franciozi, Carlos Eduardo da Silveira; Sasaki, Pedro Henrique; Abdalla, Rene Jorge

    2014-01-01

    This study is to report a manual method to obtain platelet rich plasma (PRP). For this study 61 ml of peripheral blood was obtained and submitted to centrifugation at 541g for 5 min. The centrifugation separates the blood into three components: red blood cells, buffy coat and platelet rich plasma. Blood and platelet rich plasma samples were sent to the Hospital's Laboratory and platelets and leukocytes were measured. A sample of 637 blood donors was evaluated. The platelet yield efficiency was 86.77% and the increase in platelet concentration factor was 2.89 times. The increase in leukocyte concentration factor was 1.97 times. The method described here produces leukocyte-rich and platelet-rich plasma with a high platelet and leukocyte increased factor. Level of Evidence IV, Controlled Laboratory Study.

  18. Simulation of Polymer Physical Gel With Platelet Fillers

    NASA Astrophysics Data System (ADS)

    Xu, Di; Gerssape, Dilip

    Platelet filler such as clays have superior effects on the properties of polymer gels. We used molecular dynamic simulations to study platelet filled composite gels system, in which small hexagonal disks simulate the platelets and gelation is due to short-range attraction between end-monomers and platelets. The properties of platelet filled composites are studied as a function of filler concentration. The mechanism of gelation was found similar to those of pure polymer gels; the polymers and platelets formed organic-inorganic networks, which percolate at high enough filler concentration. It was observed platelets aggregated into local intercalation structure, which significantly differs from typical spherical fillers. This unique intercalation structure is examined by radial distribution function and ordering parameters. We discussed how intercalation would affect the properties of the platelet composites by comparing them with spherical fillers.

  19. Binding of activated platelets to WBCs in vivo after transfusion.

    PubMed

    Gutensohn, Kai; Geidel, Katja; Brockmann, Marc; Siemensen, Margaux; Krueger, William; Kroeger, Nico; Kuehnl, Peter

    2002-10-01

    During preparation and storage of apheresis concentrates, platelets are being activated. One of the alterations that occur during this process is an increased expression of P-selectin (CD62p) on the cytoplasmic surface of platelets. This neoepitope represents a ligand for the binding of platelets to WBCs. It has been suggested that the activation of platelets is associated with the sequestration of platelets after transfusion. In this in vivo study, the binding of platelets to WBCs was analyzed following transfusion of platelet concentrates (PCs). Double apheresis concentrates were prepared with two different cell separators. One of the split products was stored for 1 to 2 days and the other one for 3 to 5 days. Flow cytometry was applied to analyze the degree of platelet activation in vitro, and also to measure the extent of platelet binding to WBC subclasses in vivo after transfusion into patients. The results of this study show that platelet activation occurs during apheresis and storage of PCs. After transfusion of the PCs, no significant binding of platelets to T or B-cells could be detected. However, a significant binding of platelets to monocytes and neutrophil granulocytes occurs. While in Baxter PCs stored for 1-2 days the amount of platelet-leukocyte aggregates in vivo was higher compared to COBE PCs, no such difference could be detected anymore for the PCs stored for 3-5 days. This study demonstrates that binding of activated platelets occurs to monocytes and neutrophil granulocytes but not to T- and B-cells in the circulation after transfusion. In addition, the interaction of platelets and WBCs is dependent on the degree of P-selectin expression. Platelets showing a higher degree of activation adhere to WBCs to a higher degree than nonactivated platelets.

  20. Modelling gas exchange during platelet storage without agitation.

    PubMed

    Torres, R; Tormey, C A

    2016-11-01

    The aim of this study was to create a model of oxygen distribution within platelet storage bags to evaluate implications of reduced agitation approaches. Based on our model, platelet concentration and surface area most affect internal partial pressure of oxygen, while temperature modifications have least effect, indicating primary potential approaches for optimization of platelet storage with reduced or absent agitation.

  1. Generation of functional platelets from human embryonic stem cells in vitro via ES-sacs, VEGF-promoted structures that concentrate hematopoietic progenitors.

    PubMed

    Takayama, Naoya; Nishikii, Hidekazu; Usui, Joichi; Tsukui, Hiroko; Sawaguchi, Akira; Hiroyama, Takashi; Eto, Koji; Nakauchi, Hiromitsu

    2008-06-01

    Human embryonic stem cells (hESCs) could potentially represent an alternative source for blood transfusion therapies and a promising tool for studying the ontogeny of hematopoiesis. When we cultured hESCs on either C3H10T1/2 or OP-9 cells to facilitate hematopoiesis, we found that exogenous administration of vascular endothelial growth factor promoted the emergence of sac-like structures, which we named embryonic stem cell-derived sacs (ES-sacs). These ES-sacs consisted of multiple cysts demarcated by cellular monolayers that retained some of the properties of endothelial cells. The spherical cells inside ES-sacs expressed primarily CD34, along with VE-cadherin, CD31, CD41a, and CD45, and were able to form hematopoietic colonies in semisolid culture and to differentiate into mature megakaryocytes by day 24 in the presence of thrombopoietin. Apparently, ES-sacs provide a suitable environment for hematopoietic progenitors. Relatively large numbers of mature megakaryocytes could be induced from the hematopoietic progenitors within ES-sacs, which were then able to release platelets that displayed integrin alpha IIb beta 3 activation and spreading in response to ADP or thrombin. This novel protocol thus provides a means of generating platelets from hESCs, which could serve as the basis for efficient production of platelets for clinical transfusion and studies of thrombopoiesis.

  2. Platelet activation determines the severity of thrombocytopenia in dengue infection

    PubMed Central

    Ojha, Amrita; Nandi, Dipika; Batra, Harish; Singhal, Rashi; Annarapu, Gowtham K.; Bhattacharyya, Sankar; Seth, Tulika; Dar, Lalit; Medigeshi, Guruprasad R.; Vrati, Sudhanshu; Vikram, Naval K.; Guchhait, Prasenjit

    2017-01-01

    Thrombocytopenia is common in patients with dengue virus (DENV) infections. With a focus on understanding the possible mechanism of thrombocytopenia in DENV infections we described a direct correlation between activation and depletion of platelets in patients. Our data showed a sharp decrease in platelet counts at day 4 of fever in patients. The high DENV genome copies in platelets correlated directly with the elevated platelet activation along with increased binding of complement factor C3 and IgG on their surface at day 4. Recovery in platelet count was observed on day 10 through day 6 and 8 with simultaneous decrease in platelet activation markers. Further, our in vitro data supported the above observations describing a concentration-dependent increase in platelet activation by DENV serotype-2. The high copy number of DENV2 genome in the platelet pellet correlated directly with platelet activation, microparticle generation and clot formation. Furthermore the DENV2-activated platelets were phagocytosed in large numbers by the monocytes. The DENV2-mediated lysis and clearance of platelets were abrogated in presence of platelet activation inhibitor, prostacyclin. These observations collectively suggest that platelet activation status is an important determinant of thrombocytopenia in dengue infections. A careful strategy of inactivation of platelets may rescue them from rapid destruction during DENV infections. PMID:28139770

  3. High concentration plasma-reduced plateletapheresis concentrates.

    PubMed

    Perseghin, Paolo

    2011-06-01

    Single-donor hyperconcentrated plateletapheresis (dry-platelets) collection has been introduced in the 90's as a part of the newly developed multi-component collection strategy. This approach allowed to safely collect multiple components from a single apheresis donation, i.e. RBC, FFP and/or plateletpheresis units. Dry-platelets are usually resuspended in additive solution to maintain an adequate pH during the storage period until use. Some concern existed about possible higher degrees of platelet activation in dry-platelets units when compared to standard concentration (1.0-1.6 × 10(6)/μL platelets) units and its possible correlation with lower in vivo efficiency and/or survival of the former units. Several authors investigated this specific issue, and dry-platelets units proved to be equally effective than standard concentration plateletpheresis units in recipients. The use of dry-platelets units may reduce (i) the risk of passive infusion of naturally occurring ABO-related hemolytic antibodies when donor O platelets are given to group A, B, or AB recipient, (ii) the risk of TRALI when multiparous donors undergo plateletpheresis. Furthermore, dry-platelet collection may allow for an increased amount of FFP sent to industry. Finally, hyperconcentrated platelet units may be used for "niche" indications, such as intrauterine platelet transfusion or, in case of autologous dry-platelet collection, for further freezing for long term storage in selected patients within onco-hematological settings.

  4. Platelet lysate mucohadesive formulation to treat oral mucositis in graft versus host disease patients: a new therapeutic approach.

    PubMed

    Del Fante, Claudia; Perotti, Cesare; Bonferoni, Maria Cristina; Rossi, Silvia; Sandri, Giuseppina; Ferrari, Franca; Scudeller, Luigia; Caramella, Carla Marcella

    2011-09-01

    Optimal treatment of oral mucositis (OM) due to graft versus host disease (GvHD) is currently not available. Platelet-derived growth factors (PDGFs) have high capability for tissue healing and may play a role in repairing the mucosal barrier. The aim of the present work was to develop a mucoadhesive formulation to administer platelet lysate to oral cavity prolonging contact time of platelet lysate with oral mucosa. The mucoadhesive formulation was characterized for in vitro properties (PDGF-AB concentration, mucoadhesive properties, cytotoxicity, fibroblast proliferation, wound healing). Moreover, a preliminary clinical study on seven GvHD patients with OM refractory to other therapies was conducted, to evaluate feasibility, safety, and efficacy. GVPL (mucoadhesive gel vehicle mixed with platelet lysate)showed good mucoadhesive properties; additionally, it was characterized by good biocompatibility in vitro on fibroblasts and it was able to enhance fibroblast proliferation and wound healing, maintaining the efficacy for up to 14 days following storage at 2-8°C. In vivo, clinical response was good-to-complete in five, fair in one, none in the remaining one. The in vitro results indicate that GVPL has optimal mucoadhesive and healing enhancer properties, maintained over time (up to 14 days); preliminary clinical results suggest that oral application of platelet lysate-loaded mucoadhesive formulation is feasible, safe, well tolerated, and effective. A larger controlled randomized study is needed.

  5. Human Platelet Senescence.

    DTIC Science & Technology

    1976-04-30

    electrophoresis platelet subpopulations membranes enrichment of platelet collections with miigathrombocytes 20. A DST RAC T (Cpntinue on reverse&. "~gaU...label which enters megakaryocytes but not peripheral blood platelets. Platelets re- leased from the bone marrow however, do contain the isotope . With...their own platelet-rich plasma (anticoagulated with ACD-A) at 800 RP’M) 1200 RPM, 1600 RPM, 1800 RPM and 2000 RPM in a Sorvall RC3 Centrifugue . The

  6. [In vitro platelet production].

    PubMed

    Dunois-Lardé, C; Baruch, D

    2011-04-01

    This review aims at presenting a state of the art on platelet functions, not only in well-characterized hemostasis and thrombosis, but also in various domains such as inflammation, immunity, angiogenesis, source of growth factors, metastasis and vascular remodelling. This multivalent phenotype of platelets suggests new potential applications of platelets. The second objective is to present new advances in platelet formation from megakaryocytes and direct platelet release, as initially shown by our group and more recently by others.

  7. In vitro effect of sodium nitrite on platelet aggregation in human platelet rich plasma--preliminary report.

    PubMed

    Kadan, M; Doğanci, S; Yildirim, V; Özgür, G; Erol, G; Karabacak, K; Avcu, F

    2015-10-01

    The role of nitrates and nitric oxide on platelet functions has obtained an increasing attention with respect to their potential effects on cardiovascular disorders. In this study we aimed to analyze the effect of sodium nitrite on platelet functions in human platelets. This in vitro study was designed to show the effect of sodium nitrite on platelet functions in seven healthy volunteers. Blood samples were centrifuged to prepare platelet rich plasma and platelet poor plasma. Platelet rich plasma was diluted with the platelet poor plasma to have a final count of 300,000 ± 25,000 platelets. Platelet rich plasma was incubated with six different increasing doses (from 10 μM to 5 mM) of sodium nitrite for 1 hour at 37°C. Then stimulating agents including collagen (3 μg ml-1), adenosine diphosphate (10 μM), and epinephrine (10 μM) were added to the cuvette. Changes in light transmission were observed for 10 minutes. In addition spontaneous aggregation were performed in control group with all aggregating agents separately. Effect of sodium nitrite on agonist-induced platelet aggregation depends on the concentration of sodium nitrite. Compared with control group, agonist-induced platelet aggregations were significantly suppressed by sodium nitrite at the concentration of 5, 1.0 and 0.5 mM. Our results suggested that sodium nitrite has inhibitory effects in vitro on platelet aggregation in a dose-dependent manner.

  8. Platelet factor 4 is chemotactic for neutrophils and monocytes.

    PubMed Central

    Deuel, T F; Senior, R M; Chang, D; Griffin, G L; Heinrikson, R L; Kaiser, E T

    1981-01-01

    Platelet factor 4 is shown to be a chemotactic protein for human polymorphonuclear leukocytes and monocytes at concentrations found in human serum and reached locally in injured tissue. The maximum chemotactic response to platelet factor 4 nearly equals that achieved with saturating concentrations of the chemotactic activity derived from the fifth component of human complement, C5. Cells desensitized to C5 chemotactic activity retain chemotactic responsiveness to platelet factor 4. Serum contains inhibitory capacity against the chemotactic activity associated with platelet factor 4. Our results suggest that the local release of platelet factor 4 may be an important stimulus attracting inflammatory cells to sites of blood vessel injury. PMID:6945600

  9. Biochemical and functional abnormalities in hypercholesterolemic rabbit platelets

    SciTech Connect

    Dalal, K.B.; Ebbe, S.; Mazoyer, E.; Carpenter, D.; Yee, T. )

    1990-02-01

    This study was designed to elucidate changes in rabbit platelet lipids induced by a cholesterol rich diet and to explore the possible correlation of these lipid changes with platelet abnormalities. Pronounced biochemical alterations were observed when serum cholesterol levels of 700-1000 mg% were reached. Hypercholesterolemic (HC) platelets contained 37% more neutral lipids and 16% less phospholipids than the controls. Lysolecithin, cholesterol esters and phosphatidylinositol (PI) levels were increased in HC platelets, and the levels of phosphatidylcholine (PC) were decreased. The cholesterol/phospholipid molar ratio of lipidemic platelets increased from 0.55 +/- 0.011 to 0.89 +/- 0.016 (P less than 0.01) in eight weeks. HC platelets had 90% more arachidonic acid (AA) in the PI than normal platelets. No significant changes in AA of PC were observed. Platelet function was monitored by the uptake and release of (14C)serotonin in platelet rich plasma (PRP), using varying concentrations of collagen as an aggregating agent. The uptake of (14C)serotonin in HC and normal platelets ranged from 78-94%. The percent of (14C)serotonin released from normal and HC platelets was proportional to the concentration of collagen. However, lipidemic platelets were hyperreactive to low concentrations of collagen. Incorporation of 50 microM acetylsalicylic acid into the aggregating medium suppressed the release of (14C)serotonin in normal PRP by more than 90%, but had only a partial effect on lipidemic PRP.

  10. Modulatory effect of coffee on platelet function.

    PubMed

    Bhaskar, Shobha; Rauf, Arun A

    2010-01-01

    Blood platelets play a major role in cardiovascular disease (CVD) and thrombosis. Conflicting information exists regarding the effect of coffee consumption on the cardiovascular system. We have investigated whether the consumption of moderate amount of coffee affect platelet functions and primary hemostasis in vivo in normal and high fat diet fed rats. Coffee fed group showed significant (P < 0.05) decrease in mean platelet volume, platelet crit and platelet distribution width as compared to high fat diet (HFD) group. The concentration of malondialdehyde in platelets increased in atherosclerotic group indicates the increased thromboxane A2 (TXA2) production from membrane arachidonic acid and it was decreased in coffee treated group. Platelet aggregation studies with ADP, collagen, arachidonic acid and epinephrine showed significant (P < 0.05) decrease in aggregation in coffee fed group. Scanning electron microscopic studies revealed that platelet aggregation tendency increased in HFD group and was reduced in coffee fed group. These results indicate that coffee is active in inhibiting platelet aggregation, a critical step involved in thrombosis.

  11. Design and feasibility of high temperature nanoparticle fluid filter in hybrid thermal/photovoltaic concentrating solar power

    NASA Astrophysics Data System (ADS)

    DeJarnette, Drew; Brekke, Nick; Tunkara, Ebrima; Hari, Parameswar; Roberts, Kenneth; Otanicar, Todd

    2015-09-01

    A nanoparticle fluid filter used with concentrating hybrid solar/thermal collector design is presented. Nanoparticle fluid filters could be situated on any given concentrating system with appropriate customized engineering. This work shows the design in the context of a trough concentration system. Geometric design and physical placement in the optical path was modeled using SolTrace. It was found that a design can be made that blocks 0% of the traced rays. The nanoparticle fluid filter is tunable for different concentrating systems using various PV cells or operating at varying temperatures.

  12. Feasibility Studies for Production of Pellet Grade Concentrate from Sub Grade Iron Ore Using Multi Gravity Separator

    NASA Astrophysics Data System (ADS)

    Rao, Gottumukkala Venkateswara; Markandeya, R.; Kumar, Rajan

    2017-07-01

    An attempt has been made to utilise Sub Grade Iron Ore by producing pellet grade concentrate from Deposit 5, Bacheli Complex, Bailadila, Chhattisgarh, India. The `as received' Run of Mine (ROM) sample assayed 40.80% Fe, 40.90% SiO2. Mineralogical studies indicated that the main ore mineral is Hematite and lone gangue mineral is Quartz. Mineral liberation studies indicated that, the ore mineral Hematite and gangue mineral Quartz are getting liberated below 100 microns. The stage crushed and ground sample was subjected to concentration by using a Multi Gravity Separator (MGS). Rougher Multi Gravity Separation (MGS) experimental results were optimised to recover highest possible iron values. A concentrate of 55.80% Fe with a yield of 61.73% by weight with a recovery of 84.42% Iron values was obtained in rougher MGS concentrate. Further experiments were carried out with rougher MGS concentrate to produce a concentrate suitable for commercial grade pellet concentrate. It was proved that a concentrate assaying 66.67% Fe, 3.12% SiO2 with an yield of 45.08% by weight and with a recovery of 73.67% iron values in the concentrate.

  13. Inhibitory effect of trichothecene mycotoxins on bovine platelets stimulated by platelet activating factor.

    PubMed Central

    Gentry, P A; Ross, M L; Bondy, G S

    1987-01-01

    Several species of fungi, which infect cereals and grains, can produce a class of compounds, known as trichothecene mycotoxins, which is characterized by a substituted epoxy-trichothecene ring structure. Cattle are susceptible to intoxication from feeds contaminated with T-2 toxin, one of the more potent trichothecene mycotoxins, while swine refuse to ingest feed contaminated with T-2 toxin. The bovine platelet has been used as a model cell system to evaluate the effects of T-2 toxin and its natural metabolites, HT-2 toxin and T-2 tetraol, on cell function in vitro. Due to the lipophilic nature of these mycotoxins, a biologically active phospholipid was used to stimulate the platelets in the presence and absence of the toxins. The mycotoxin T-2 toxin and its major metabolite HT-2 toxin inhibited platelet activating factor-stimulated bovine platelets, suspended in homologous plasma, in a concentration but not time dependent manner. Significant inhibition of platelet function (p less than 0.01) occurred with 135 ng T-2 toxin per 10(6) platelets and with 77 ng HT-2 toxin per 10(6) platelets. These mycotoxins exerted an additive inhibitory effect on the platelet aggregation response. In contrast, the minor metabolite T-2 tetraol had no inhibitory effect on platelet function and had no influence on the responses of T-2 toxin or HT-2 toxin when the mycotoxins were present together in the platelet suspensions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:3453270

  14. Decrease in platelet activating factor stimulated phosphoinositide turnover during storage of human platelets in plasma

    SciTech Connect

    Carter, M.G.; Shukla, S.D. )

    1987-05-01

    Human platelet concentrate from the American Red Cross Blood Center was stored at 24{degree}C in a shaker and aliquots were taken out at time intervals aseptically. Platelet activating factor (PAF) stimulated turnover of phosphoinositide (PPI) was monitored by assaying {sup 32}P incorporation into phosphoinositides using platelet rich plasma (PRP). Platelets in PRP were incubated with 1 {times} 10{sup {minus}7} M PAF at 37{degree}C with gentle shaking and after 5 min their lipids were extracted and analysed by TLC for {sup 32}P-phosphoinositides. The percent stimulation of {sup 32}P incorporation by PAF (over control) into PPI was approximately 250, 100, 60, 25 and 20 on days 1, 2, 3, 5 and 6, respectively. This indicated a dramatic decrease in PAF responsive turnover of platelet PPI during storage. These findings have important implications in relation to PAF receptor activity and viability of platelets at different periods of storage.

  15. Functional platelet defects in children with severe chronic ITP as tested with 2 novel assays applicable for low platelet counts.

    PubMed

    van Bladel, Esther R; Laarhoven, Annemieke G; van der Heijden, Laila B; Heitink-Pollé, Katja M; Porcelijn, Leendert; van der Schoot, C Ellen; de Haas, Masja; Roest, Mark; Vidarsson, Gestur; de Groot, Philip G; Bruin, Marrie C A

    2014-03-06

    Immune thrombocytopenia (ITP) is an autoimmune disease with a complex heterogeneous pathogenesis and a bleeding phenotype that is not necessarily correlated to platelet count. In this study, the platelet function was assessed in a well-defined cohort of 33 pediatric chronic ITP patients. Because regular platelet function test cannot be performed in patients with low platelet counts, 2 new assays were developed to determine platelet function: first, the microaggregation test, measuring in platelets isolated from 10 mL of whole blood the platelet potential to form microaggregates in response to an agonist; second, the platelet reactivity assay, measuring platelet reactivity to adenosine diphosphate (ADP), convulxin (CVX), and thrombin receptor activator peptide in only 150 μL of unprocessed whole blood. Patients with a severe bleeding phenotype demonstrated a decreased aggregation potential upon phorbol myristate acetate stimulation, decreased platelet degranulation following ADP stimulation, and a higher concentration of ADP and CVX needed to activate the glycoprotein IIbIIIa complex compared with patients with a mild bleeding phenotype. In conclusion, here we have established 2 functional tests that allow for evaluation of platelet function in patients with extremely low platelet counts (<10(9)). These tests show that platelet function is related to bleeding phenotype in chronic ITP.

  16. The Role of Platelet Membrane Phospholipids in the Platelet Release Reaction

    PubMed Central

    Schick, Paul K.; Yu, Byung P.

    1974-01-01

    The structure and function of the platelet surface was probed by phospholipase C (Clostridium perfringens) which hydrolyzes membrane phospholipids, particularly phosphatidylcholine. Platelet phospholipids were susceptible to phospholipase C, and extent of hydrolysis was dependent on concentration of phospholipase C and Ca++. Phospholipase C (0.15 U/ml) with Ca++ (0.55 mM) hydrolyzed 15.6% phospholipids during 5 min. Phospholipase C released platelet serotonin (5HT), ADP, and platelet factor 4. Hydrolysis of 5% phospholipids resulted in release of 70% 5HT. Platelet 5HT release was rapid, occurring within 2 min. Phospholipase C (0.2 U/ml) with Ca++ (0.55 mM) also released 10.35 nmol sotrage pool ADP/109 platelets and 63% platelet factor 4 during 3 min. Phospholipase C did not cause leakage of cytoplasmic metabolic pool ADP, since only 6.6% [3H]ADP was released. Ultrastructural analysis of phospholipase C-modified platelets showed that platelets were intact. After 2% phospholipid hydrolysis, centralization of granules and contraction of microtubules were evident. After 18% phospholipid hydrolysis, there were morphological indications of degranulation. Phospholipase C-induced phospholipid hydrolysis caused the release of ADP and 5HT since: (a) Phospholipase C purified by heating was shown to be free of protease and neuraminidase activity and capable of inducing the platelet release reaction. (b) Antitoxin (Cl. perfringens) neutralized phospholipase C-induced 5HT release which rules out a contaminant. (c) Phosphorylcholine, the hydrolysis product, did not induce platelet 5HT release. This study demonstrates that minimal hydrolysis of platelet phospholipids triggers the release reaction. Our hypothesis is that phospholipids, presumably phosphatidylcholine, are situated at or near active site or “receptor” on the platelet surface and function as the modulator for the release reaction. Images PMID:4371563

  17. Feasibility of estimating heavy metal concentrations in water column using hyperspectral data and partial least squares regression

    NASA Astrophysics Data System (ADS)

    Chen, Yiyun; Liu, Yaolin; Wang, Dun; Kong, XueSong; Zeng, Chen

    2009-10-01

    Mining and smelting often produce acidic wastes that can cause severe biogeochemical changes downstream from these mines. Dexing copper mine, as the largest open cast mine in China, is connected to Poyang Lake by Le An river. Water and spectra samples were taken from Le An River and two of its branches, and afterward the concentrations of Cd, Cu, Pb and Zn were measured in the lab. Different spectral pre-processing methods were applied to the spectra, including Savitzky-Golay spectral smoothing, SNV, first derivative, second derivative spectral transforming. On the purpose of estimating metal concentrations from differently pre-processed spectra, partial least squares regression was then used in model calibrations. For deciding the optimal number of PLS factors included in the PLS model, the model with the lowest root mean square error of validation is chosen. The coefficient of determination (R2v) between the predicted and the reference values from the test set are used as an evaluation mean. For estimating Pb concentration, R2v = 0.915, which is acceptable. For Cd concentration, R2v = 0.697 and 0.683 for Zn. PLS model seems to failed in estimating Cu concentration, for the best R2v for PLS model of Cu is lower than 0.5. From the aspects of spectral pre-processing methods, first derivative after Savitzky-Golay smoothing performs superior to others. In conclusion, PLS models based on carefully pre-processed hyperspectral data turn out to be a promising solution for detecting certain heavy metals concentrations in river.

  18. Clinical application of radiolabelled platelets

    SciTech Connect

    Kessler, C. )

    1990-01-01

    This book presents papers on the clinical applications of radiolabelled platelets. The papers are grouped into six sections on platelet labelling techniques, radiolabelled platelets in cardiology, monitoring of antiplatelet therapy, platelet scintigraphy in stroke patients, platelet scintigraphy in angiology, and platelet scintigraphy in hematology and other clinical applications, including renal transplant rejection.

  19. Platelet granule release is associated with reactive oxygen species generation during platelet storage: A direct link between platelet pro-inflammatory and oxidation states.

    PubMed

    Ghasemzadeh, Mehran; Hosseini, Ehteramolsadat

    2017-08-01

    Upon platelet stimulation with agonists, reactive oxygen species (ROS) generation enhances platelet activation and granule release. Whether ROS generation during platelet storage could be directly correlated with the expression of proinflammatory molecules and granule release has been investigated in this study. PRP-platelet concentrates were subjected to flowcytometry analysis to assess the expression of platelet activation marker, P-selectin and CD40L during storage. Intracellular ROS generation was also detected in platelet by flowcytometry using dihydrorhodamine (DHR) 123. Through the dual staining, ROS production was analyzed in either P-selectin positive or negative populations. ROS formation in platelet population was significantly increased by either TRAP (a potent agonist that induces granule release) or PMA (a classic inducer of ROS generation), while the effects of each agonists on P-selectin expression and ROS generation in platelets were comparable. Platelet storage was also associated with the increasing levels of ROS (day 0 vs. day 5; p<0.001) while this increasing pattern was directly correlated with the either expressed P-selectin or CD40L. In addition, in 5 day-stored platelets, samples with ROS levels above 40% showed significantly higher levels of P-selectin and CD40L expression. P-selectin negative population of platelet did not show significant amount of ROS. Our data demonstrated decreased levels of important platelet pro-inflammatory molecules in stored platelets with lower levels of intraplatelet ROS. However, whether quenching of ROS generation during platelet storage can attenuate adverse transfusion reactions raised by platelet pro-inflammatory status is required to be further studied. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Guidance on platelet transfusion for patients with hypoproliferative thrombocytopenia.

    PubMed

    Nahirniak, Susan; Slichter, Sherrill J; Tanael, Susano; Rebulla, Paolo; Pavenski, Katerina; Vassallo, Ralph; Fung, Mark; Duquesnoy, Rene; Saw, Chee-Loong; Stanworth, Simon; Tinmouth, Alan; Hume, Heather; Ponnampalam, Arjuna; Moltzan, Catherine; Berry, Brian; Shehata, Nadine

    2015-01-01

    Patients with hypoproliferative thrombocytopenia are at an increased risk for hemorrhage and alloimmunization to platelets. Updated guidance for optimizing platelet transfusion therapy is needed as data from recent pivotal trials have the potential to change practice. This guideline, developed by a large international panel using a systematic search strategy and standardized methods to develop recommendations, incorporates recent trials not available when previous guidelines were developed. We found that prophylactic platelet transfusion for platelet counts less than or equal to 10 × 10(9)/L is the optimal approach to decrease the risk of hemorrhage for patients requiring chemotherapy or undergoing allogeneic or autologous transplantation. A low dose of platelets (1.41 × 10(11)/m2) is hemostatically as effective as higher dose of platelets but requires more frequent platelet transfusions suggesting that low-dose platelets may be used in hospitalized patients. For outpatients, a median dose (2.4 × 10(11)/m2) may be more cost-effective to prevent clinic visits only to receive a transfusion. In terms of platelet products, whole blood-derived platelet concentrates can be used interchangeably with apheresis platelets, and ABO-compatible platelet should be given to improve platelet increments and decrease the rate of refractoriness to platelet transfusion. For RhD-negative female children or women of child-bearing potential who have received RhD-positive platelets, Rh immunoglobulin should probably be given to prevent immunization to the RhD antigen. Providing platelet support for the alloimmunized refractory patients with ABO-matched and HLA-selected or crossmatched products is of some benefit, yet the degree of benefit needs to be assessed in the era of leukoreduction.

  1. Transcellular activation of platelets and endothelial cells by bioactive lipids in platelet microparticles.

    PubMed Central

    Barry, O. P.; Pratico, D.; Lawson, J. A.; FitzGerald, G. A.

    1997-01-01

    , which it may then use to synthesize PGI2. Both PGE2 and iloprost, a stable PGI2 analog, evoke human umbilical vein endothelial cell COX-2 expression, albeit with kinetics that differ from the response to platelet microparticles. These studies indicate a novel mechanism of transcellular lipid metabolism whereby platelet activation may be amplified or modulated by concentrated delivery of arachidonic acid to adjacent platelets and endothelial cells. PMID:9151784

  2. Loss of 111Indium as indicator of platelet injury

    SciTech Connect

    Joist, J.H.; Baker, R.K.

    1981-08-01

    We previously demonstrated that platelets can be labeled with 111Inoxine with high labeling efficiency and that 111In is not liberated from labeled platelets during the platelet release reaction or prolonged in vitro storage. In view of these findings, we examined the potential usefulness of loss of 111In from labeled platelets as an indicator of platelet damage by comparing the loss of 111In with that of 51Cr and LDH (in some experiments also with platelet factor 3 availability) under different conditions of platelet injury. When washed human platelets labeled with either 51Cr-chromate or 111In-oxine were exposed to increasing concentrations of detergents (Triton X-100, lysolecithin), threshold, rate, and extent of loss of 111In, 51Cr and, LDH were similar. In contrast, when labeled platelets were depleted of metabolic energy by incubation in glucose-free Tyrode albumin solution or glucose-depleted plasma in the presence of antimycin A and 2-deoxy-D-glucose, loss of 51Cr (and PF3a) occurred earlier and progressed at a faster rate than that of 111In or LDH. Similar results were obtained when platelets were exposed to increasing concentrations of PlA1 antibody, causing complement-mediated immune injury. The findings indicate that with certain agents that cause rapid platelet disruption (lysis), different platelet constituents are lost at similar rates. However, under conditions of more subtle or slowly progressive platelet injury, small molecules such as adenine nucleotides (51Cr) may escape earlier and at faster rates than larger molecules such as LDH or 111In-binding platelet protein. Thus, neither 111In loss nor LDH loss appear to be suitable indicators for sublytic or prelytic platelet injury.

  3. Platelets contribute to growth and metastasis in hepatocellular carcinoma.

    PubMed

    Bihari, Chhagan; Rastogi, Archana; Shasthry, Saggere Muralikrishna; Bajpai, Meenu; Bhadoria, Ajeet Singh; Rajesh, S; Mukund, Amar; Kumar, Anupam; Sarin, Shiv K

    2016-09-01

    To determine the association of platelets with hepatocellular carcinoma (HCC) growth and its metastasis. We examined platelets, laboratory, and radiological data of consecutive 420 HCC and 1008 cirrhosis cases. Follow-up information of platelet count in cirrhosis to HCC, pre- to post-therapy, and post-therapy to HCC outcome was analyzed. Cytokine profiling was performed in HCC and cirrhosis (n = 10 each). On the basis of imaging, HCC was divided into six subgroups. Cytosmears of HCC were assessed for platelet clustering around tumor cells. An in vitro Matrigel invasion assay was performed on human HCC cell lines using graded concentration of platelets. Baseline platelet numbers and platelet/lymphocyte ratios (PLRs) were significantly higher (p < 0.001) in HCC than cirrhosis. IL-1, IL-6, FGF, G-CSF, thrombopoietin, and VEGF were higher in HCC than cirrhosis. Platelet counts were increased after HCC conversion of cirrhosis (p < 0.001) and decreased (p < 0.001) after therapy. Platelets and PLR in recurrence cases were higher than in responders at baseline. AFP, PIVKAII, platelets, and PLR increase (p < 0.001 each) with advancement in HCC growth. Multivariate analysis showed platelets (p = 0.002), PLR (p = 0.004), and AFP (p < 0.001) associated with distant metastasis. Platelet clustering seen in 75.7% of HCC group 3, 45% in group 2, and 12.5% in group 1 cases (p < 0.001). Invaded cells in Matrigel assay positively correlated with platelet concentration. Platelets can contribute to the development, growth, invasion, and metastasis of HCC. Rising platelet count after HCC therapy is indicative of incomplete response or recurrence.

  4. The effect of desmopressin on platelet function: a selective enhancement of procoagulant COAT platelets in patients with primary platelet function defects.

    PubMed

    Colucci, Giuseppe; Stutz, Monika; Rochat, Sophie; Conte, Tiziana; Pavicic, Marko; Reusser, Marianne; Giabbani, Evelyne; Huynh, Anh; Thürlemann, Charles; Keller, Peter; Alberio, Lorenzo

    2014-03-20

    1-deamino-8-d-arginine vasopressin (desmopressin [DDAVP]) is clinically efficacious in patients with mild platelet function disorders but it is not known which mechanisms mediate this effect. Our aim was to evaluate the impact of in vivo DDAVP administration in these patients. We assessed von Willebrand factor (VWF), factor VIII, platelet activation and aggregation, platelet-dependent thrombin generation, and platelet intracellular Na(+)/Ca(2+) fluxes, before and 2 and 4 hours after DDAVP (0.3 µg/kg). We found (1) no significant changes for P-selectin expression, PAC-1 binding, δ-granule content and secretion, and platelet-aggregation; (2) significant decreases of secretion of α-granules and GPIIb-IIIa activation induced by adenosine 5'-diphosphate, convulxin, and thrombin; (3) significant increases of procoagulant platelets induced by convulxin/thrombin and platelet-dependent thrombin generation; and (4) significant increases of intracellular Na(+)/Ca(2+) concentrations. We show that in vivo DDAVP selectively and markedly enhances the ability to form procoagulant platelets and increases platelet-dependent thrombin generation by enhancing Na(+)/Ca(2+) mobilization. This report indicates that the beneficial hemostatic effect of DDAVP is not limited to an increase in large VWF multimers. An enhancement of platelet procoagulant activity appears to be an additional and (at least in platelet disorders) -possibly clinically relevant mechanism of DDAVP's action.

  5. Diltiazem potentiates the inhibitory effect of aspirin on platelet aggregation.

    PubMed

    Altman, R; Scazziota, A; Dujovne, C

    1988-09-01

    Platelet aggregation in vivo occurs through the combined effects of many agonists. Aspirin inhibits platelet aggregation but its antiaggregate effects can be overcome by the synergistic action of sodium arachidonate (AA) plus platelet activating factor (PAF). We tested the effect of a calcium entry-blocking agent, diltiazem, on AA-PAF-induced platelet aggregation in platelet-rich plasma from seven healthy volunteers. The studies were done before and after aspirin (100 mg/day) administration for 7 to 10 days. Stimulation of platelet was done in vitro by AA, PAF, or both. Before aspirin treatment, diltiazem (2 micrograms/ml) added in vitro to the platelet-rich plasma inhibited platelet aggregation induced by AA (0.75 mmol/L) by 50%. When PAF was used the inhibition of aggregation was obtained at a lower concentration of diltiazem (0.4 to 1 microgram/ml). After aspirin treatment, AA-induced aggregation was inhibited, and PAF alone (30 nmol/L) produced a first-wave aggregation followed by complete disaggregation. When AA and PAF were added together a full aggregation of postaspirin treatment platelets was obtained. Diltiazem added in vitro at the clinically attainable concentration of 0.1 microgram/ml produced a complete inhibition of this AA-PAF synergism on platelet aggregation. These results suggest that administration of a combination of low-dose aspirin and diltiazem may be of greater benefit than aspirin alone for prophylaxis of cardiovascular diseases where platelets are involved in the pathogenesis.

  6. Labile aggregation stimulating substance, free fatty acids, and platelet aggregation.

    PubMed

    Gerrard, J M; White, J G; Krivit, W

    1976-01-01

    Labile aggregation stimulating substance (LASS), an intermediate produced during platelet biosynthesis of PGE2 and PGF2alpha, acts as a physiologic intercellular messenger to promote platelet aggregation and the release reaction. The activity is formed by intact cells after physiologic stimulation or can be generated from platelet membrane fractions after combination with arachidonate. In the present investigation, small amounts of polyunsaturated fatty acids added to an incubation mixture of platelet microsomes and arachidonate were found to significantly inhibit subsequent platelet aggregation. Saturated and mono-unsaturated fatty acids in the same concentrations were without effect. However, in higher concentrations mono-unsaturated fatty acids were found to be inhibitory and stearic acid was found to enhance subsequent platelet aggregation. The inhibition caused by the polyunsaturated fatty acid, linoleate, was shown to be the result of an effect on the production of LASS through an interaction with the platelet enzyme responsible for conversion of arachidonate to LASS. In contrast, stearic acid was found to enhance platelet aggregation by acting on the platelets and not directly on LASS production. The results suggest that small changes in the fatty acid composition of platelet phospholipids could significantly influence platelet reactivity.

  7. The omnipotent platelet.

    PubMed

    Steinberg, L A

    1996-03-01

    This information was derived from the increase in platelets of patients following fractures and/or bone surgery and in conjunction with a vast amount of published literature. The increase in numbers of platelets reflects the extent of bone involvement, especially noted in the hip, knee, post-coronary artery bypass graft, and multiple fractures. The role of the platelet in any and all tissues, i.e. soft tissue or bone, whether beneficial or detrimental, is multifunctional. The platelet responds to all physiologic and pathologic states and, if tissue involved is sufficient, the role of the platelet becomes obvious.

  8. Cangrelor attenuates coated-platelet formation.

    PubMed

    Norgard, Nicholas B; Hann, Callie L; Dale, George L

    2009-01-01

    P2Y(12) inhibitors were introduced clinically as effective inhibitors of adenosine-5'-diphosphate (ADP) mediated platelet activation and aggregation. This class of pharmacological agents has enjoyed considerable success. Cangrelor is a recently developed P2Y(12) inhibitor that has the advantage of being an active drug not requiring metabolic conversion, although it is not orally available. Coated-platelets are a subclass of activated platelets generated on dual agonist activation with collagen plus thrombin; the primary hallmark of coated-platelets is their ability to support prothrombinase activity. Interestingly, we recently observed that the relatively weak agonist ADP potentiates the production of coated-platelets by the very strong agonists collagen plus thrombin, a previously unknown role for ADP. The authors sought in this study to determine if P2Y(12) inhibitors, such as cangrelor, were capable of attenuating this augmentation of coated-platelet generation. Cangrelor, at physiologically relevant concentrations, was able to eliminate the ADP-dependent increase in coated-platelet production with an IC(50) of 1.4 nM. Cangrelor, however, had no effect on thrombin-dependent platelet activation as measured by P-selectin expression. Although this in vitro study does not address the question of whether the effectiveness of cangrelor in vivo is partially due to an attenuation of coated-platelet production in addition to its documented antiaggregatory effects, it does reveal an unexpected action of cangrelor. Additional studies will be required to determine if all P2Y(12) inhibitors are equally effective in attenuating coated-platelet production.

  9. Rhesus monkey platelets

    SciTech Connect

    Harbury, C.B.

    1986-03-01

    The purpose of this abstract is to describe the adenine nucleotide metabolism of Rhesus monkey platelets. Nucleotides are labelled with /sup 14/C-adenine and extracted with EDTA-ethanol (EE) and perchlorate (P). Total platelet ATP and ADP (TATP, TADP) is measured in the Holmsen Luciferase assay, and expressed in nanomoles/10/sup 8/ platelets. TR=TATP/TADP. Human platelets release 70% of their TADP, with a ratio of released ATP/ADP of 0.7. Rhesus platelets release 82% of their TADP, with a ratio of released ATP/ADP of 0.33. Thus, monkey platelets contain more ADP than human platelets. Thin layer chromatography of EE gives a metabolic ratio of 11 in human platelets and 10.5 in monkey platelets. Perchlorate extracts metabolic and actin bound ADP. The human and monkey platelets ratios were 5, indicating they contain the same proportion of actin. Thus, the extra ADP contained in monkey platelets is located in the secretory granules.

  10. Human anterior cruciate ligament fibroblasts from immature patients have a stronger in vitro response to platelet concentrates than those from mature individuals

    PubMed Central

    Magarian, Elise M.; Vavken, Patrick; Murray, Martha M.

    2010-01-01

    A number of recently published studies have established a substantial age dependence of the response of ACL fibroblasts to stimulation by platelet-rich plasma (PRP). Further in-depth research of this age dependence revealed negative effects on both histological and biomechanical results in a large animal model. However, while it has been postulated that this association could affect potential human applications negatively too it remains to be proven that the same effects occur in human cells. Thus it was the objective of this study to search for age dependence in human fibroblasts before further human experiments are done. Human fibroblasts were obtained from 10 immature and adolescent patients, based on a-priori power calculations, and culture in a collagen-PRP composite. Three parameters that are pivotal for defect remodeling and wound healing – cell migration, cell proliferation, and scaffold contraction – were chosen as endpoints. Both migration and proliferation were significantly higher in immature cells, but no differences were seen in wound contraction. The former findings suggest that immature patients respond more favorably to treatment with PRP, which consequently might translate into better results in ACL tissue engineering. PMID:20728363

  11. Adsorption of methylene blue using Fe3O4/CuO/ZnO/ nanographene platelets (NGP) composites with various NGP concentration

    NASA Astrophysics Data System (ADS)

    Tju, H.; Taufik, A.; Saleh, R.

    2016-11-01

    This study will examine the use of Fe3O4/CuO/ZnO nanocomposites that have been modified by Nanographene Platelets (NGP) as an adsorbent to degrade organic dye waste Methylene Blue (MB). The nanocomposites were synthesized using the sol-gel method then combined with three variatons of NGP weight percents by simple hydrothermal method. The Fe3O4/CuO/ZnO/NGP composites were characterized using the X-Ray Diffraction (XRD) spectroscopy, Fourier Transform Infrared (FTIR), Energy Dispersive X-Ray (EDX), Thermogravimetric Analysis (TGA) and Vibrating Sample Magnetometer (VSM). The composites exhibit ferromagnetic behaviour. The presence of hexagonal wurtzite of ZnO, monoclinic of CuO and cubic spinel of Fe3O4 were found in the composites. The graphitic-like structure represents the presence of the NGP in the composites. However, the addition of NGP weight percent reduces the thermal stability of the composites. The adsorption capability of the composites are analyzed by observing the degradation of organic dye Methylene Blue (MB) under dark condition. The NGP addition of 15 wt% show the best result of the composites to degrade Methylene Blue in alkaline condition. Adsorption mechanism of the composites with NGP addition tend to follow the model Langmuir adsorption kinetic models.

  12. Aspirin treatment reduces platelet resistance to deformation.

    PubMed

    Burris, S M; Smith, C M; Rao, G H; White, J G

    1987-01-01

    The present investigation has evaluated the influence of aspirin, its constituents, and other nonsteroidal anti-inflammatory agents on the resistance of human platelets to aspiration into micropipettes. Aspirin increased the length of platelet extensions into the micropipette over the entire negative tension range of 0.04 to 0.40 dynes/cm after exposure to the drug in vitro or after ingestion of the agent. Other cyclooxygenase inhibitors, ibuprofen and indomethacin, did not increase platelet deformability. The influence of aspirin was mimicked to some degree by high concentrations of salicylic acid, but acetylation of platelets with acetic anhydride had little influence on platelet deformability. Incubation of platelets with both salicylic acid and acetic anhydride had no more effect than salicylic acid alone. Benzoic acid, chemically similar to salicylic acid, had a minimal effect. The studies demonstrate that aspirin makes platelets more deformable, while components of the drug or other nonsteroidal antiinflammatory agents and cyclooxygenase inhibitors do not have the same influence on resistance to deformation.

  13. Improving platelet transfusion safety: biomedical and technical considerations

    PubMed Central

    Garraud, Olivier; Cognasse, Fabrice; Tissot, Jean-Daniel; Chavarin, Patricia; Laperche, Syria; Morel, Pascal; Lefrère, Jean-Jacques; Pozzetto, Bruno; Lozano, Miguel; Blumberg, Neil; Osselaer, Jean-Claude

    2016-01-01

    Platelet concentrates account for near 10% of all labile blood components but are responsible for more than 25% of the reported adverse events. Besides factors related to patients themselves, who may be particularly at risk of side effects because of their underlying illness, there are aspects of platelet collection and storage that predispose to adverse events. Platelets for transfusion are strongly activated by collection through disposal equipment, which can stress the cells, and by preservation at 22 °C with rotation or rocking, which likewise leads to platelet activation, perhaps more so than storage at 4 °C. Lastly, platelets constitutively possess a very large number of bioactive components that may elicit pro-inflammatory reactions when infused into a patient. This review aims to describe approaches that may be crucial to minimising side effects while optimising safety and quality. We suggest that platelet transfusion is complex, in part because of the complexity of the “material” itself: platelets are highly versatile cells and the transfusion process adds a myriad of variables that present many challenges for preserving basal platelet function and preventing dysfunctional activation of the platelets. The review also presents information showing - after years of exhaustive haemovigilance - that whole blood buffy coat pooled platelet components are extremely safe compared to the gold standard (i.e. apheresis platelet components), both in terms of acquired infections and of immunological/inflammatory hazards. PMID:26674828

  14. Platelet function, activation and apoptosis during and after apheresis.

    PubMed

    Bakry, Rania; Sayed, Douaa; Galal, Hanan; Shaker, Sanaa

    2010-10-01

    Platelets are known to undergo shape change, activation, release reaction and apoptosis/necrosis during processing and storage. Apheresis may have a deleterious impact on platelet achievability and functional integrity. Platelet concentrates from 50 male volunteers obtained by COBE spectra were screened for platelet activation (CD62 and CD154) and apoptosis (phosphatidylserine detected by Annexin V). Donor samples before separation, during apheresis and at the third day of storage were used as baseline donor samples. Platelet aggregation to adenosine diphosphate (ADP) and collagen was performed. There was a statistically significant increase in the expression of activation markers in two different samples (during separation samples and third day samples). Although the increase in Annexin V expression was not so observable, it showed a significant increase also. There was marked decline in the platelet aggregation. The correlations between the values of CD62, CD154 and Annexin V were detected in baseline samples and increased during separation and at the third day of platelets storage. Correlation between values of platelet aggregation to collagen and Annexin V was relevant only in the baseline samples. No other correlations were encountered between platelet aggregation and markers of activation and apoptosis during apheresis and storage. Initial platelet activation induced by apheresis may have an impact on phosphatidylserine expression with no impact on aggregation function of platelets during storage.

  15. Glutamate mediates platelet activation through the AMPA receptor

    PubMed Central

    Morrell, Craig N.; Sun, Henry; Ikeda, Masahiro; Beique, Jean-Claude; Swaim, Anne Marie; Mason, Emily; Martin, Tanika V.; Thompson, Laura E.; Gozen, Oguz; Ampagoomian, David; Sprengel, Rolf; Rothstein, Jeffrey; Faraday, Nauder; Huganir, Richard; Lowenstein, Charles J.

    2008-01-01

    Glutamate is an excitatory neurotransmitter that binds to the kainate receptor, the N-methyl-D-aspartate (NMDA) receptor, and the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor (AMPAR). Each receptor was first characterized and cloned in the central nervous system (CNS). Glutamate is also present in the periphery, and glutamate receptors have been identified in nonneuronal tissues, including bone, heart, kidney, pancreas, and platelets. Platelets play a central role in normal thrombosis and hemostasis, as well as contributing greatly to diseases such as stroke and myocardial infarction. Despite the presence of glutamate in platelet granules, the role of glutamate during hemostasis is unknown. We now show that activated platelets release glutamate, that platelets express AMPAR subunits, and that glutamate increases agonist-induced platelet activation. Furthermore, we demonstrate that glutamate binding to the AMPAR increases intracellular sodium concentration and depolarizes platelets, which are important steps in platelet activation. In contrast, platelets treated with the AMPAR antagonist CNQX or platelets derived from GluR1 knockout mice are resistant to AMPA effects. Importantly, mice lacking GluR1 have a prolonged time to thrombosis in vivo. Our data identify glutamate as a regulator of platelet activation, and suggest that the AMPA receptor is a novel antithrombotic target. PMID:18283118

  16. Indium-111-labeled platelet scintigraphy in carotid atherosclerosis

    SciTech Connect

    Minar, E.; Ehringer, H.; Dudczak, R.; Schoefl, R.J.; Jung, M.; Koppensteiner, R.; Ahmadi, R.; Kretschmer, G.

    1989-01-01

    We evaluated platelet accumulation in carotid arteries by means of a dual-radiotracer method, using indium-111-labeled platelets and technetium-99m-labeled human serum albumin, in 123 patients (92 men, 31 women; median age 60 years). Sixty patients had symptoms of transient ischemic carotid artery disease, and 63 patients with peripheral arterial occlusive disease served as controls. Antiplatelet treatment with acetylsalicylic acid was taken by 53 of the 123 patients. In 36 of the 60 symptomatic patients, platelet scintigraphy was repeated 3-4 days after carotid endarterectomy. Comparison of different scintigraphic parameters (platelet accumulation index and percent of the injected dose of labeled platelets at the carotid bifurcation) showed no significant differences between symptomatic and asymptomatic patients, and the severity of stenosis and the presence of plaque ulceration also had no influence on the parameters. There was no difference between patients with a short (less than 4 weeks) or long (greater than 4 weeks) interval from the last transient ischemic attack to scintigraphy and no difference between patients with or without antiplatelet treatment. Classifying the patients according to plaque morphology judged by high-resolution real-time ultrasonography also demonstrated no differences. No significant correlation was found between any scintigraphic parameter and other platelet function parameters such as platelet survival time, platelet turnover rate, and concentration of platelet-specific proteins. Quantification of platelet deposition after carotid endarterectomy in 36 patients demonstrated a significant increase of the median platelet accumulation index and the percent injected dose index.

  17. Storage of platelet concentrates from pooled buffy coats made of fresh and overnight-stored whole blood processed on the novel Atreus 2C+ system: in vitro study.

    PubMed

    Sandgren, Per; Callaert, Martine; Shanwell, Agneta; Gulliksson, Hans

    2008-04-01

    The Atreus 2C+ system (Gambro BCT) automatically separates whole blood (WB) into buffy coat (BC), red blood cells (RBC), and plasma and transfers the components into separate containers. After processing with the Atreus, 4 to 6 BC units can be pooled and processed into leukoreduced platelets (PLTs) by use of the automated OrbiSac BC system (Gambro BCT). The aim of our in vitro study was to investigate the effects of holding either WB or BC overnight before preparation of PLTs by use of the Atreus 2C+ system for BC preparation. A standard routine procedure involving conventional blood containers for the preparation of BC combined with the OrbiSac process (top-and-top system; Terumo) was used as a reference. WB was either processed within 8 hours after collection ("fresh blood") or stored overnight before processing. WB units were separated into BC, RBC, and plasma units and transferred into individual containers. Either the BC or the WB units rested overnight at 22 +/- 2 degrees C. Six ABO-identical BCs, obtained from either fresh or overnight-stored WB, were pooled and processed with the OrbiSac BC system to obtain leukoreduced PLTs. In total, 20 Atreus and 10 reference (leukoreduced PLTs) samples were analyzed for various in vitro variables during the 7-day storage period. No significant difference in glucose consumption, lactate production, mean PLT volume, LDH activity, bicarbonate, ATP, RANTES, and the expression of CD62p and CD42b between groups was detected. pH was maintained at greater than 7.0 (Day 7). Swirling remained at the highest levels (score, 2) for all units throughout storage. PLTs derived from BCs, obtained from either fresh or overnight-stored WB processed on the novel automated Atreus 2C+ system, were equivalent to control PLTs with regard to PLT in vitro characteristics during 7 days of storage. Stable recovery of PLTs and satisfactory PLT content according to current standards were also found.

  18. Assessment of quality of platelets preserved in plasma and platelet additive solution: A Malaysian experience

    PubMed Central

    Mokhtar, Munirah Binti; Hashim, Hasna Binti; Joshi, Sanmukh R

    2016-01-01

    Background: A use of platelet additives solution (PAS) improves storage conditions so as to give increased shelf life to platelets and to maintain hemostatic function. Objective: The present study was aimed to compare in vitro quality of platelet rich plasma (PRP)-derived platelet concentrate (PC) during extended period of storage in plasma and in additive solution (Composol PS and Fresenius). Study Design: Randomized 19 PCs each were used in the study for plasma and PAS as the storage medium. The measurement parameters, including pH, total white blood cell (WBC) count, total platelet count, and platelet activation rate, were studied on day 1, day 5, and day 8 of the storage period. The sterility test was carried out on the eighth day of storage. Results: pH of PC suspended in PAS was significantly lower as compared to that in plasma (P < 0.001) for all the three days of sampling. The WBC count, both in plasma and in PAS, showed an acceptable values of being <0.2 Χ 109 /unit during the storage period. Platelet count in PAS was higher as compared to that in plasma, though it was not statistically significant. While both the groups showed increased platelet activation rate during the storage, the PCs suspended in PAS showed significantly higher platelet activation rate (p0.001). Results from sterility test showed no bacterial growth in the PCs in both the groups. Conclusion: Most parameters studied on platelet storage in suspending medium of native plasma and PAS remained well within the acceptable limits. However, the pH values and platelet activation rate significantly differed in PAS as compared with plasma. PMID:27011678

  19. Feasibility of Single Scan for Simultaneous Evaluation of Regional Krypton and Iodine Concentrations with Dual-Energy CT: An Experimental Study.

    PubMed

    Hong, Sae Rom; Chang, Suyon; Im, Dong Jin; Suh, Young Joo; Hong, Yoo Jin; Hur, Jin; Kim, Young Jin; Choi, Byoung Wook; Lee, Hye-Jeong

    2016-11-01

    Purpose To evaluate the feasibility of a simultaneous single scan of regional krypton and iodine concentrations by using dual-energy computed tomography (CT). Materials and Methods The study was approved by the institutional animal experimental committee. An airway obstruction model was first made in 10 beagle dogs, and a pulmonary arterial occlusion was induced in each animal after 1 week. For each model, three sessions of dual-energy CT (80% krypton ventilation [krypton CT], 80% krypton ventilation with iodine enhancement [mixed-contrast agent CT], and iodine enhancement [iodine CT]) were performed. Krypton maps were made from krypton and mixed-contrast agent CT, and iodine maps were made from iodine and mixed-contrast agent CT. Observers measured overlay Hounsfield units of the diseased and contralateral segments on each map. Values were compared by using the Wilcoxon signed-rank test. Results In krypton maps of airway obstruction, overlay Hounsfield units of diseased segments were significantly decreased compared with those of contralateral segments in both krypton and mixed-contrast agent CT (P = .005 for both). However, the values of mixed-contrast agent CT were significantly higher than those of krypton CT for both segments (P = .005 and .007, respectively). In iodine maps of pulmonary arterial occlusion, values were significantly lower in diseased segments than in contralateral segments for both iodine and mixed-contrast agent CT (P = .005 for both), without significant difference between iodine and mixed-contrast agent CT for both segments (P = .126 and .307, respectively). Conclusion Although some limitations may exist, it might be feasible to analyze regional krypton and iodine concentrations simultaneously by using dual-energy CT. (©) RSNA, 2016.

  20. Assessment of the correlation of platelet morphology with in vivo recovery and survival.

    PubMed

    Mintz, Paul D; Anderson, Garth; Avery, Nancy; Clark, Pamela; Bonner, Robert F

    2005-08-01

    There is continuing interest in the development of in vitro tests evaluating the in vivo function, recovery, and survival of platelets stored for transfusion. A recent forum concluded that no completely reliable test exists, although discoid morphology indicates a platelet's good health. We evaluated a novel device, the NAPSAC (Noninvasive Assessment of Platelet Shape and Concentration), designed to determine noninvasively the proportion of discoid platelets in a stored concentrate, as well as platelet concentration. Twenty-eight plateletapheresis concentrates stored 24 hours in PL-146 were evaluated. Percent discoid platelet results were correlated with radiolabeled autologous recovery and survival performed using 111Indium oxyquinoline and calculated using linear (L) and multiple-hit (M) models. pH of 8 concentrates was raised at the end of storage with 6N NaOH. Platelet concentration measured by NAPSAC and Coulter Thrombocounter C was compared in 256 plateletapheresis products. Percent discoid platelets at 24 hours did not correlate significantly with platelet recovery or survival (recovery L = 0.29, M = 0.28; survival L = 0.16, M = 0.03). Raising the pH (mean 6.38 to 6.94) resulted in a significant increase in percent discoid platelets (21% to 41%). Platelet concentration values for both methods studied were linearly correlated with a slope of 1.01 +/- 0.03, r = 0.81. Percent discoid platelets was not predictive of posttransfusion platelet recovery or survival. The results suggest that non-discoid platelets may survive posttransfusion and even revert to discoid shape, since raising the pH approximately doubled the percent of discoid platelets. The NAPSAC was shown to be a reliable instrument for noninvasively determining platelet concentration in PL-146 concentrates.

  1. Assessment of platelet function in healthy sedated cats using three whole blood platelet function tests.

    PubMed

    Ho, Kimberly K; Abrams-Ogg, Anthony C G; Wood, R Darren; O'Sullivan, M Lynne; Kirby, Gordon M; Blois, Shauna L

    2015-05-01

    The objectives of this study were to establish feline references intervals for 3 commercial whole blood platelet function test analyzer systems: Multiplate analyzer (MP; Roche Diagnostics International Ltd., Rotkreuz, Switzerland), Platelet Function Analyzer-100 (PF: Siemens Canada, Mississauga, Ontario, Canada), and Plateletworks Combo-25 kit (PW; Helena Laboratories, Beaumont, TX). Venipuncture was performed on 55 healthy sedated cats, and platelet aggregation in response to adenosine diphosphate (ADP), collagen (COL), and arachidonic acid (AA; MP only) was assessed using citrated blood. For the MP analyzer, median (95% confidence intervals [CIs]) area under curve (Units) for ADP, COL, and AA agonists were 87 (11-176), 81 (32-129), and 91 (59-129), respectively. For the PF analyzer, median (95% CIs) closure time, using COL-ADP cartridges, was 69 (46-89) sec. For the PW assay, median (95% CIs) percent aggregations for ADP and COL agonists were 71 (18-92) and 49 (9-96), respectively, using impedance hematology analyzer platelet counts, and 94 (25-98) and 68 (14-119), respectively, using flow cytometry hematology analyzer platelet counts. There were low correlations between the PF analyzer (COL-ADP cartridge) and MP analyzer (COL agonist; ρ = 0.11), and between the PF analyzer (COL-ADP cartridge) and PW assay (COL agonist using impedance platelet counts; ρ = 0.14). The PW assay percent aggregations using impedance and flow cytometric platelet counts were correlated for both ADP (ρ = 0.64) and COL (ρ = 0.64) agonists. Platelet function testing using these tests are feasible in cats, but 95% CIs are wide, so single results may be difficult to interpret. Platelet counting by impedance or flow cytometry may be used for the PW assay but are not interchangeable.

  2. Polyphenols enhance platelet nitric oxide by inhibiting protein kinase C-dependent NADPH oxidase activation: effect on platelet recruitment.

    PubMed

    Pignatelli, P; Di Santo, S; Buchetti, B; Sanguigni, V; Brunelli, A; Violi, F

    2006-06-01

    Several studies demonstrated an inverse association between polyphenol intake and cardiovascular events. Platelet recruitment is an important phase of platelet activation at the site of vascular injury, but it has never been investigated whether polyphenols influence platelet recruitment. The aim of the study was to analyze in vitro whether two polyphenols, quercetin and catechin, were able to affect platelet recruitment. Platelet recruitment was reduced by NO donors and by NADPH oxidase inhibitors and was enhanced by L-NAME, an inhibitor of NO synthase. Quercetin and catechin, but not single polyphenol, significantly inhibited platelet recruitment in a concentration-dependent fashion. The formation of superoxide anion was significantly inhibited in platelets incubated with quercetin and catechin but was unaffected by a single polyphenol. Incubation of platelets with quercetin and catechin resulted in inhibition of PKC and NADPH oxidase activation. Treatment of platelets with quercetin and catechin resulted in an increase of NO and also down-regulated the expression of GpIIb/IIIa glycoprotein. This study shows that the polyphenols quercetin and catechin synergistically act in reducing platelet recruitment via inhibition of PKC-dependent NADPH oxidase activation. This effect, resulting in NO-mediated platelet glycoprotein GpIIb/IIIa down-regulation, could provide a novel mechanism through which polyphenols reduce cardiovascular disease.

  3. Thrombotic thrombocytopenic purpura: a syndrome of intravascular platelet consumption.

    PubMed Central

    Neame, P. B.; Hirsh, J.; Browman, G.; Denburg, J.; D'Souza, T. J.; Gallus, A.; Brain, M. C.

    1976-01-01

    In four of five patients with thrombotic thrombocytopenic purpura (TTP) in whom serial tests of hemostatic function were performed, severe thrombocytopenia, normal plasma fibrinogen concentrations and mildly increased concentrations of fibrinogen/fibrin degradation products were observed. Widespread platelet thrombi were found in arterioles and capillaries. Fibrin could be seen around some of the platelet clumps and was the main component in a small number of the thrombi in two patients. The observations show that TTP is a disorder in which intravascular platelet consumption results in disseminated platelet thrombosis. The coagulation system is apparently activated secondarily to platelet aggregation and variable quantities of fibrin are incorporated into the thrombi. Clinical improvement resulted from combined therapy with corticosteroids, heparin and drugs that suppress platelet function. Images FIG. 3 FIG. 4 FIG. 5 FIG. 6 PMID:1084215

  4. Antioxidants change platelet responses to various stimulating events

    PubMed Central

    Sobotková, Alžběta; Mášová-Chrastinová, Leona; Suttnar, Jiří; Štikarová, Jana; Májek, Pavel; Reicheltová, Zuzana; Kotlín, Roman; Weisel, John W.; Malý, Martin; Dyr, Jan E.

    2010-01-01

    The role of platelets in hemostasis may be influenced by alteration of the platelet redox state—the presence of antioxidants and the formation of reactive oxygen and nitrogen species. We investigated the effects of two antioxidants, resveratrol and trolox, on platelet activation. Trolox and resveratrol inhibited aggregation of washed platelets and platelet-rich plasma activated by ADP, collagen, and thrombin receptor-activating peptide. Resveratrol was a more effective agent in reducing platelet static and dynamic adhesion in comparison with trolox. The antioxidant capacity of resveratrol was, however, the same as that of trolox. After incubation of platelets with antioxidants, the resveratrol intraplatelet concentration was about five times lower than the intracellular concentration of trolox. Although both antioxidants comparably lowered hydroxyl radical and malondialdehyde production in platelets stimulated with collagen, TxB2 levels were decreased by resveratrol much more effectively than by trolox. Cyclooxygenase 1 was inhibited by resveratrol and not by trolox. Our data indicate that antioxidants, apart from nonspecific redox or radical-quenching mechanisms, inhibit platelet activation also by specific interaction with target proteins. The results also show the importance of studying platelet activation under conditions of real blood flow in contact with reactive surfaces, e.g., using dynamic adhesion experiments. PMID:19766712

  5. Feasibility of low-concentration iodinated contrast medium with lower-tube-voltage dual-source CT aortography using iterative reconstruction: comparison with automatic exposure control CT aortography.

    PubMed

    Shin, Hee Jeong; Kim, Song Soo; Lee, Jae-Hwan; Park, Jae-Hyeong; Jeong, Jin-Ok; Jin, Seon Ah; Shin, Byung Seok; Shin, Kyung-Sook; Ahn, Moonsang

    2016-06-01

    To evaluate the feasibility of low-concentration contrast medium (CM) for vascular enhancement, image quality, and radiation dose on computed tomography aortography (CTA) using a combined low-tube-voltage and iterative reconstruction (IR) technique. Ninety subjects underwent dual-source CT (DSCT) operating in dual-source, high-pitch mode. DSCT scans were performed using both high-concentration CM (Group A, n = 50; Iomeprol 400) and low-concentration CM (Group B, n = 40; Iodixanol 270). Group A was scanned using a reference tube potential of 120 kVp and 120 reference mAs under automatic exposure control with IR. Group B was scanned using low-tube-voltage (80 or 100 kVp if body mass index ≥25 kg/m(2)) at a fixed current of 150 mAs, along with IR. Images of the two groups were compared regarding attenuation, image noise, signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), iodine load, and radiation dose in various locations of the CTA. In comparison between Group A and Group B, the average mean attenuation (454.73 ± 86.66 vs. 515.96 ± 101.55 HU), SNR (25.28 ± 4.34 vs. 31.29 ± 4.58), and CNR (21.83 ± 4.20 vs. 27.55 ± 4.81) on CTA in Group B showed significantly greater values and significantly lower image noise values (18.76 ± 2.19 vs. 17.48 ± 3.34) than those in Group A (all Ps < 0.05). Homogeneous contrast enhancement from the ascending thoracic aorta to the infrarenal abdominal aorta was significantly superior in Group B (P < 0.05). Low-concentration CM and a low-tube-voltage combination technique using IR is a feasible method, showing sufficient contrast enhancement and image quality.

  6. The influence of prostaglandin G2 on platelet ultrastructure and platelet secretion.

    PubMed Central

    Gerrard, J. M.; Townsend, D.; Stoddard, S.; Witkop, C. J.; White, J. G.

    1977-01-01

    Prostaglandin G2 (PGG2) is a labile endoperoxide produced physiologically following exposure of platelets to aggregating agents. We report here studies using isolated PGG2. This agent stimulates a concentration-dependent internal platelet contraction very similar to that produced by the calcium ionophore A23187. EDTA prevented platelet aggregation but did not prevent PGG2-stimulated internal contraction or secretion. In contrast, prostaglandin E1 and dibutyryl cyclic AMP inhich selectively labilizes platelet granules, was added to platelets together with PGG2 there was a superadditive effect on platelet secretion. Thus, granule labilization induced by PMA is a separable phenomenon and complementary to the effect of PGG2 on contraction. The ultimate degree of secretion is dependent on both processes. Studies using additional inhibitors supported the hypothesis that PGG2 activates platelets (either directly or following conversion to thromboxane A2) by transporting calcium from an intracellular store to the cytoplasmic site of the platelet contractile proteins. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 PMID:188341

  7. Redox Proteomics and Platelet Activation: Understanding the Redox Proteome to Improve Platelet Quality for Transfusion

    PubMed Central

    Sonego, Giona; Abonnenc, Mélanie; Tissot, Jean-Daniel; Prudent, Michel; Lion, Niels

    2017-01-01

    Blood banks use pathogen inactivation (PI) technologies to increase the safety of platelet concentrates (PCs). The characteristics of PI-treated PCs slightly differ from those of untreated PCs, but the underlying reasons are not well understood. One possible cause is the generation of oxidative stress during the PI process. This is of great interest since reactive oxygen species (ROS) act as second messengers in platelet functions. Furthermore, there are links between protein oxidation and phosphorylation, another mechanism that is critical for cell regulation. Current research efforts focus on understanding the underlying mechanisms and identifying new target proteins. Proteomics technologies represent powerful tools for investigating signaling pathways involving ROS and post-translational modifications such as phosphorylation, while quantitative techniques enable the comparison of the platelet resting state versus the stimulated state. In particular, redox cysteine is a key player in platelet activation upon stimulation by different agonists. This review highlights the experiments that have provided insights into the roles of ROS in platelet function and the implications for platelet transfusion, and potentially in diseases such as inflammation and platelet hyperactivity. The review also describes the implication of redox mechanism in platelet storage considerations. PMID:28208668

  8. Umbilical Cord Blood Platelet Lysate as Serum Substitute in Expansion of Human Mesenchymal Stem Cells.

    PubMed

    Shirzad, Negin; Bordbar, Sima; Goodarzi, Alireza; Mohammad, Monire; Khosravani, Pardis; Sayahpour, Froughazam; Baghaban Eslaminejad, Mohamadreza; Ebrahimi, Marzieh

    2017-10-01

    The diverse clinical applications for human mesenchymal stem cells (hMSCs) in cellular therapy and regenerative medicine warrant increased focus on developing adequate culture supplements devoid of animal-derived products. In the present study, we have investigated the feasibility of umbilical cord blood-platelet lysate (UCB-PL) as a standard substitute for fetal bovine serum (FBS) and human peripheral blood-PL (PB-PL). In this experimental study, platelet concentrates (PC) from UCB and human PB donors were frozen, melted, and sterilized to obtain PL. Quality control included platelet cell counts, sterility testing (viral and microbial), total protein concentrations, growth factor levels, and PL stability. The effects of UCB-PL and PB-PL on hMSCs proliferation and differentiation into osteocytes, chondrocytes, and adipocytes were studied and the results compared with FBS. UCB-PL contained high levels of protein content, platelet-derived growth factor- AB (PDGF-AB), and transforming growth factor (TGF) compared to PB-PL. All growth factors were stable for at least nine months post-storage at -70˚C. hMSCs proliferation enhanced following treatment with UCB-PL. With all three supplements, hMSCs could differentiate into all three lineages. PB-PL and UCB-PL both were potent in hMSCs proliferation. However, PB promoted osteoblastic differentiation and UCB-PL induced chondrogenic differentiation. Because of availability, ease of use and feasible standardization of UCB-PL, we have suggested that UCB-PL be used as an alternative to FBS and PB-PL for the cultivation and expansion of hMSCs in cellular therapy.

  9. Analysis of aggregation of platelets in thrombosis

    NASA Astrophysics Data System (ADS)

    Ahuja, Suresh

    Platelets are key players in thrombus formation by first rolling over collagen bound von Willebrand factor followed by formation of a stable interaction with collagen. The first adhered platelets bind additional platelets until the whole injury is sealed off by a platelet aggregate. The coagulation system stabilizes the formed platelet plug by creating a tight fibrin network, and then wound contraction takes place because of morphological changes in platelets. Coagulation takes place by platelet activation and aggregation mainly through fibrinogen polymerization into fibrin fibers. The process includes multiple factors, such as thrombin, plasmin, and local shear-rate which regulate and control the process. Coagulation can be divided into two pathways: the intrinsic pathway and the extrinsic pathway. The intrinsic pathway is initiated by the exposure of a negatively charged. It is able to activate factor XII, using a complex reaction that includes prekallikrein and high-molecular-weight kininogen as cofactors.. Thrombin is the final enzyme that is needed to convert fibrinogen into fibrin. The extrinsic pathway starts with the exposure of tissue factor to the circulating blood, which is the major initiator of coagulation. There are several feedback loops that reinforce the coagulation cascade, resulting in large amounts of thrombin. It is dependent on the presence of pro-coagulant surfaces of cells expressing negatively charged phospholipids--which include phosphatidylserine (PS)--on their outer membrane. PS-bearing surfaces are able to increase the efficiency of the reactions by concentrating and co-localizing coagulation factors.. Aggregation of platelets are analyzed and compared to adhesion of platelet to erythrocyte and to endothelial cells. This abstract is replacing MAR16-2015-020003.

  10. Functional fractionation of platelets.

    PubMed

    Haver, V M; Gear, A R

    1981-02-01

    Studies of platelet populations suggest that they are heterogeneous in size, age, and metabolic parameters. In an attempt to correlate these parameters with efficiency of aggregation, a new technique, functional fractionation, was developed. Platelet populations are separated by their differential reactivity to aggregating agents. For example, low doses of ADP (0.1 to 0.7 microM) are added to stirred PRP, after which gentle centrifugation is used to remove aggregates from single unreacted platelets. The loose aggregates can be readily dispersed for comparison of the physical or biochemical properties of the reacted versus unreacted platelets. It was found that reactive platelets were larger (6.5 micrometer3) than unreacted platelets (5.51 micrometer3). No significant difference in density existed between the two populations, and no release of [14C]serotonin from prelabeled platelets occurred during functional fractionation. Scanning and transmission electron microscopy confirmed the size difference and revealed that in both populations platelets were structurally intact with a normal discoid shape and no significant difference in organelle content. Human platelets most reactive to ADP were also enriched in glycogen (3.6-fold), ATP (1.6-fold), and ADP (twofold), compared with less reactive cells. These "reactive" cells took up more 51[Cr] and contained 1.9 times more surface sialic acid. In an in vivo aging experiment, rats were injected with 75[Se]methionine. Shortly after labeling (1 day), the most reactive platelets possessed the highest amount of 75[Se]. These results reveal that functionally active platelets, which are also larger, are more active metabolically than less reactive platelets, possess a higher negative surface charge, and may be a younger population.

  11. Endotoxin Interactions with Platelets

    DTIC Science & Technology

    1985-01-01

    irreversible aggregation of human platelets (Hamberg and Sainuelsson 1974; Hlamberg et al 1975). Acetylsalicylic acid , an inhibitor of cyclooxygenase aud...exposure to endotoxin (100 ttg/nil). To simulate the lipopolysac- charide of endotoxin, several different fatty acids were added individually to platelet...platelet</