Congenital platelet function defects
Platelet storage pool disorder; Glanzmann's thrombasthenia; Bernard-Soulier syndrome; Platelet function defects - congenital ... This disorder may also cause severe bleeding. Platelet storage pool disorder (also called platelet secretion disorder) occurs ...
Studies of Platelet 5-Hydroxytryptamine (Serotonin) in Storage Pool Disease and Albinism
Weiss, Harvey J.; Tschopp, Thomas B.; Rogers, John; Brand, Harvey
1974-01-01
Platelets in patients with storage pool disease are markedly deficient in a nonmetabolic (storage) pool of ADP that is important in platelet aggregation. They are also deficient in ATP, although to a lesser degree. In seven patients with this disorder, including one with albinism, platelet 5-hydroxytryptamine (5-HT) levels were reduced in proportion to the reduction in ATP (r = 0.94). Their platelets show diminished capacity to absorb [14C]5-HT, and the type of defect was similar to that produced in normal platelets by reserpine, a drug known to inhibit the uptake of 5-HT by the platelet dense granules. Storage pool-deficient platelets also converted more [3H]5-HT to [3H]5-hydroxyindoleacetic acid than did normal platelets, and the platelets in one of two patients studied contained increased amounts of 5-HT metabolites. The above findings, together with those reported previously, support the conclusion that the capacity of the dense granules (which may be either diminished or functionally abnormal) for storing 5-HT is decreased in storage pool disease; as a result, the 5-HT that enters the platelet may be more exposed to monoamine oxidases present on mitochondrial membranes. This diminished storage capacity (for 5-HT) may also explain why preincubating platelet-rich plasma with 5-HT for 45 min without stirring inhibits subsequent platelet aggregation by 5-HT to a greater degree in patients with storage pool disease than in normal subjects. The latter finding is also consistent with the theory that the aggregation of platelets by 5-HT is mediated by the same receptors on the plasma membrane that are involved in its uptake. The diminished release of platelet-bound [14C]5-HT by collagen that we found in these patients, as well as findings in previous studies, suggests that the release reaction may also be abnormal in storage pool disease. Images PMID:4847252
Storage of platelets: effects associated with high platelet content in platelet storage containers.
Gulliksson, Hans; Sandgren, Per; Sjödin, Agneta; Hultenby, Kjell
2012-04-01
A major problem associated with platelet storage containers is that some platelet units show a dramatic fall in pH, especially above certain platelet contents. The aim of this study was a detailed investigation of the different in vitro effects occurring when the maximum storage capacity of a platelet container is exceeded as compared to normal storage. Buffy coats were combined in large-volume containers to create primary pools to be split into two equal aliquots for the preparation of platelets (450-520×10(9) platelets/unit) in SSP+ for 7-day storage in two containers (test and reference) with different platelet storage capacity (n=8). Exceeding the maximum storage capacity of the test platelet storage container resulted in immediate negative effects on platelet metabolism and energy supply, but also delayed effects on platelet function, activation and disintegration. Our study gives a very clear indication of the effects in different phases associated with exceeding the maximum storage capacity of platelet containers but throw little additional light on the mechanism initiating those negative effects. The problem appears to be complex and further studies in different media using different storage containers will be needed to understand the mechanisms involved.
Alhumaidan, Hiba; Cheves, Tracey; Holme, Stein; Sweeney, Joseph D
2011-10-01
The processing of whole blood-derived platelet-rich plasma (PRP) to a platelet concentrate and platelet-poor plasma is currently performed within 8 hours to comply with the requirements to manufacture fresh frozen plasma. Maintaining PRP at room temperature for a longer period can have the advantage of shifting the completion of component manufacture onto day shifts. Pairs of ABO-identical prepooled platelets were manufactured by the PRP method, using the current approach with platelet storage in a CLX HP container (Pall Medical, Covina, CA) and plasma, or a novel approach with an 18- to a 24-hour room temperature hold of the PRP and the manufacture of pooled platelets in a glucose-containing additive solution (AS) and storage in a new ELX container (Pall Medical). Standard in vitro assays were performed on days 2, 5, and 7. The results showed that the AS platelets in ELX have in vitro characteristics that are equivalent or superior to those of the standard product.
Maximising platelet availability by delaying cold storage.
Wood, B; Johnson, L; Hyland, R A; Marks, D C
2018-04-06
Cold-stored platelets may be an alternative to conventional room temperature (RT) storage. However, cold-stored platelets are cleared more rapidly from circulation, reducing their suitability for prophylactic transfusion. To minimise wastage, it may be beneficial to store platelets conventionally until near expiry (4 days) for prophylactic use, transferring them to refrigerated storage to facilitate an extended shelf life, reserving the platelets for the treatment of acute bleeding. Two ABO-matched buffy-coat-derived platelets (30% plasma/70% SSP+) were pooled and split to produce matched pairs (n = 8 pairs). One unit was stored at 2-6°C without agitation (day 1 postcollection; cold); the second unit was stored at 20-24°C with constant agitation until day 4 then stored at 2-6°C thereafter (delayed-cold). All units were tested for in vitro quality periodically over 21 days. During storage, cold and delayed-cold platelets maintained a similar platelet count. While pH and HSR were significantly higher in delayed-cold platelets, other metabolic markers, including lactate production and glucose consumption, did not differ significantly. Furthermore, surface expression of phosphatidylserine and CD62P, release of soluble CD62P and microparticles were not significantly different, suggesting similar activation profiles. Aggregation responses of delayed-cold platelets followed the same trend as cold platelets once transferred to cold storage, gradually declining over the storage period. The metabolic and activation profile of delayed-cold platelets was similar to cold-stored platelets. These data suggest that transferring platelets to refrigerated storage when near expiry may be a viable option for maximising platelet inventories. © 2018 International Society of Blood Transfusion.
Prüller, F; Rosskopf, K; Mangge, H; Mahla, E; von Lewinski, D; Weiss, E C; Riegler, A; Enko, D
2017-12-01
Essentials In platelet function testing, standardized internal controls (IQC) are not commercially provided. Platelet function testing was performed daily on aliquoted pooled platelet concentrates. Pooled platelet concentrates showed stability for control purposes from Monday to Friday. Pooled platelet concentrates provide the necessary steadiness to serve as IQC material. Background Standardized commercially available control material for internal quality control (IQC) of light transmission aggregometry (LTA) is still lacking. Moreover, the availability of normal blood donors to provide fresh platelets is difficult in small laboratories, where 'volunteers' may be in short supply. Objectives To evaluate the implementation of buffy-coat-derived pooled platelet concentrates (PCs) for IQC material for LTA. Methods We used buffy-coat-derived pooled PCs from the blood bank as IQC material for LTA. On each weekend one PC was prepared (> 200 mL) and aliquoted from the original storage bag on a daily basis in four baby bags (40-50 mL), which were delivered from Monday to Friday to our laboratory. The IQC measurements of at least 85 work-weeks (from Monday to Friday) were evaluated with this new IQC material. LTA was performed on a four-channel Chronolog 700 Aggregometer (Chronolog Corporation, Havertown, PA, USA) (agonists: collagen, adenosine diphosphate [ADP], arachidonic acid [AA] and thrombin receptor activator peptide-6 [TRAP-6]). Results The medians of platelet aggregation from IQC measurements with collagen, ADP and AA from Monday to Friday were 68.0-59.5, 3.0-2.0 and 51.0-50.0%, respectively, and the mean of platelet aggregation with TRAP-6 was 71.2-66.4%. Conclusions Buffy-coat-derived pooled PCs serve as a reliable and robust IQC material for LTA measurements and would be beneficial for the whole laboratory procedure and employees' safety. © 2017 International Society on Thrombosis and Haemostasis.
Saunders, C; Rowe, G; Wilkins, K; Holme, S; Collins, P
2011-08-01
The non-paired two-arm study compared the in vitro storage characteristics of platelets suspended as concentrates in either 100% plasma or a mixture of additive solution (SSP+™, MacoPharma, Mouveaux, France) and autologous plasma in a 70:30 ratio over a 14-day storage period. The buffy coat-derived pooled platelet concentrates were sampled on days 1, 2, 3, 6, 8, 10 and 14 and tests performed to determine platelet morphology, function, metabolism, activation and apoptosis-like activity. Swirling remained strong (score=3) in SSP+™, whilst scores of 1 and 0 were noted for plasma units by end of storage. In contrast to units in plasma, pH levels remained above seven in SSP+™ units, increasing after day 10. Percent positive expression of CD62P was similar in both groups on day 1 (median of 54% and 56% for plasma (n=13) and SSP+™ (n=12), respectively), with SSP+™ units showing a more moderate increase in activation after day 10. A progressive decrease in mitochondrial membrane potential was evident in both groups from day 1, whilst annexin V binding was relatively stable from days 1 to 3, with median values remaining below 6%. Subsequent to this, the percentage of platelets binding annexin V increased to approximately 30% by day 14. Platelets suspended in a medium of 70:30 SSP+™ to plasma ratio performed at least as well as platelets in 100% autologous plasma for up to 10 days of storage. Further, results are suggestive of an apoptosis-like process being involved in the platelet storage lesion. © 2011 The Author(s). Vox Sanguinis © 2011 International Society of Blood Transfusion.
Johnson, Lacey; Tan, Shereen; Wood, Ben; Davis, April; Marks, Denese C
2016-07-01
Alternatives to room temperature storage of platelets (PLTs) may be beneficial to extend the limited shelf life and support transfusion logistics in rural and military areas. The aim of this study was to assess the morphologic, metabolic, and functional aspects of PLTs stored at room temperature or in refrigerated conditions or cryopreserved. A three-arm pool-and-split study was carried out using buffy coat-derived PLTs stored in 30% plasma/70% SSP+. The three matched treatment arms were room temperature stored (20-24°C), cold-stored (2-6°C), and cryopreserved (-80°C with dimethyl sulfoxide). Liquid-stored PLTs were tested over a 21-day period, while cryopreserved PLTs were examined immediately after thawing and after 6 and 24 hours of storage at room temperature. Cold-stored and cryopreserved PLTs underwent a significant shape change, although the cryopreserved PLTs appeared to recover from this during subsequent storage. Glycolytic metabolism was reduced in cold-stored PLTs, but accelerated in cryopreserved PLTs, while oxidative phosphorylation was negatively affected by both storage conditions. PLT aggregation was potentiated by cold storage and diminished by cryopreservation in comparison to room temperature-stored PLTs. Cold storage and cryopreservation resulted in faster clot formation (R-time; thromboelastography), which was associated with an increase in microparticles. Cold storage and cryopreservation of PLTs led to morphologic and metabolic changes. However, storage under these conditions appears to maintain or even enhance certain aspects of in vitro PLT function. © 2016 AABB.
Sireis, W; Rüster, B; Daiss, C; Hourfar, M K; Capalbo, G; Pfeiffer, H-U; Janetzko, K; Goebel, M; Kempf, V A J; Seifried, E; Schmidt, M
2011-10-01
The Paul-Ehrlich-Institute analysed all fatalities due to bacterial infections between 1997 and 2007. Thereafter, the platelet shelf life was reduced to a maximum of 4 days after blood donation because the majority of all cases of severe transfusion-transmitted bacterial infections occurred with day 5 platelets. The current study compares the analytical sensitivity and the diagnostic specificity of four rapid bacterial detection procedures. Nine transfusion-relevant bacterial strains were spiked in pooled platelets or apheresis platelets at a low concentration (10 CFU/bag). Samples were collected after day 3, day 4 and day 5 and investigated by four rapid bacterial detection methods (modified BacT/ALERT, Bactiflow, FACS method and 16s DNA PCR methods). Seven out of nine bacterial strains were adequately detected by BacT/ALERT, Bactiflow and PCR in apheresis platelets and pooled platelets after sample collection at day 3, day 4 and day 5. For three bacterial strains, analytical sensitivity was reduced for the FACS method. Two bacterial strains did not grow under the storage conditions in either pooled or apheresis platelets. A late sample collection on day 3, day 4 or day 5 after blood donation in combination with a rapid bacterial detection method offers a new opportunity to improve blood safety and reduce errors due to sampling., BacT/ALERT, Bactiflow or 16s ID-NAT are feasible for late bacterial screening in platelets may provide data which support the extension of platelet shelf life in Germany to 5 days. © 2011 The Author(s). Vox Sanguinis © 2011 International Society of Blood Transfusion.
Improving platelet transfusion safety: biomedical and technical considerations
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
Improved extension of platelet storage in a polyolefin container with higher oxygen permeability.
Yuasa, Takeshi; Ohto, Hitoshi; Yasunaga, Reiko; Kai, Takanori; Shirahama, Noriaki; Ogata, Takashi
2004-07-01
This study evaluated a newly developed polyolefin bag, which has a 50%/m(2) higher oxygen permeability to extend the shelf-life of platelets. Single-donor aphaeresis platelets were pooled, separated equally into two bags, PO-80 (0.8 l capacity) and KBP-PO (1 l capacity) for control, and stored in plasma for up to 7 d. Platelet biochemical and functional parameters were monitored in bags containing high (4.2 x 10(11)/250 ml/bag, n = 9) and low (2.0 x 10(11)/200 ml/bag, n = 3) concentrations of platelets over the storage period. After 7 d of storage, the PO-80 bags containing high concentrations of platelets had a better pH (mean pH 6.74) than those stored in KBP-PO (pH 6.32, P < 0.01); none of the nine PO-80 bags with a high platelet concentration had a pH below 6.2, compared with four of nine controls (P < 0.05). Similarly, lactate values were 20.19 mmol/l and 28.09 mmol/l respectively (P < 0.05). Aerobic metabolism was maintained better with greater O(2) consumption and less lactate generation in high-platelet concentration PO-80 bags than in the control bags. A significant difference was also found in pH, pCO(2) and lactate levels between the two bags containing low concentrations of platelets during the 7-d period. The in vitro characteristics of platelets declined less over 7 d when stored in a higher oxygen permeable container than in a marketed bag.
Sellberg, Felix; Berglund, Erik; Ronaghi, Martin; Strandberg, Gabriel; Löf, Helena; Sommar, Pehr; Lubenow, Norbert; Knutson, Folke; Berglund, David
2016-12-01
Platelet lysate is a readily available source of growth factors, and other mediators, which has been used in a variety of clinical applications. However, the product remains poorly standardized and the present investigation evaluates the composition of platelet lysate obtained from either fresh or stored pathogen-inactivated platelet units. Platelet pooled units (n = 10) were obtained from healthy blood donors and tested according to standard procedures. All units were pathogen inactivated using amotosalen hydrochloride and UVA exposure. Platelet lysate was subsequently produced at two separate time-points, either from fresh platelet units or after 5 days of storage, by repeated freeze-thaw cycles. The following mediators were determined at each time-point: EGF, FGF-2, VEGF, IGF-1, PDGF-AB/BB, BMP-2, PF4, TGF-β isoform 1, IL-1β, IL-2, IL-6, IL-10, IL-12p70, 1L-17A, TNF-α, and IFN-γ. The concentration of growth factors and cytokines was affected by time in storage. Notably, TGF-β, PDGF-AB/BB, and PF4 showed an increase of 27.2% (p < 0.0001), 29.5% (p = 0.04) and 8.2% (p = 0.0004), respectively. A decrease was seen in the levels of IGF-1 and FGF-2 with 22% (p = 0.041) and 11% (p = 0.01), respectively. Cytokines were present only in very low concentrations and all other growth factors remained stable with time in storage. The composition of mediators in platelet lysate obtained from pathogen-inactivated platelet units differs when produced from fresh and stored platelet units, respectively. This underscores the need for further standardization and optimization of this important product, which potentially may influence the clinical effects. Copyright © 2016. Published by Elsevier Ltd.
Rinalducci, Sara; Zolla, Lello
2015-06-01
The mechanisms responsible for the reduced lifespan of transfused red blood cells (RBCs) and platelets (PLTs) are still under investigation, however one explanation refers to the detrimental biochemical changes occurring during ex vivo storage of these blood products. A myriad of historical and more recent studies has contributed to advance our understanding of storage lesion. Without any doubts, proteomics had great impact on transfusion medicine by profiling the storage-dependent changes in the total detectable protein pool of both RBCs and PLTs. This review article focuses on the role of oxidative/nitrosative stress in developing RBC and PLT storage lesions, with a special glance at its biochemistry and cross-talk with phosphorylative signal transduction. In this sense, we enlighten the potential contribution of new branches of proteomics in identifying novel points of intervention for the improvement of blood product quality. Copyright © 2015 Elsevier Ltd. All rights reserved.
Saunders, C; Rowe, G; Wilkins, K; Collins, P
2013-07-01
Glucose and acetate have been proposed to be required elements in platelet storage media. This study investigated the role of these compounds on the varied elements that comprise the platelet storage lesion. For each replicate, four pooled and split ABO group-specific buffy coat-derived platelet concentrates were suspended in an in-house additive solution with minimal plasma and varying final concentrations of acetate or glucose. Units were sampled on days 2, 3, 6, 8 and 10 and tested for markers of platelet morphology, activation, function, metabolism and indicators of cell death. The absence of glucose was associated with a decrease in ATP, falling to a mean of 1·1 ± 0·1 μmol/10(11) plts in units with no added glucose compared with 4·2 ± 0·6 μmol/10(11) plts (P < 0·001) in units with 30 mm glucose. As glucose became depleted, the decrease in ATP to levels below 3 μmol/10(11) plts was associated with an increase in both annexin V binding and intracellular free calcium. In units lacking exogenous acetate, ATP levels on day 10 were 5·2 ± 1·5 μmol/10(11) plts compared with 2·7 ± 0·9 μmol/10(11) plts in units with 56 mm acetate (P = 0·006). Higher concentrations of exogenous acetate were associated with a lower hypotonic shock response and higher surface expression of CD62P suggestive of a dose dependency. Under current physical storage conditions, glucose appears necessary for the maintenance of platelets stored as concentrates in minimal volumes of plasma. The addition of acetate was associated with increased platelet activation and reduced ATP levels. © 2013 International Society of Blood Transfusion.
Platelet storage in Fresenius/NPBI polyolefin and BTHC-PVC bags: a direct comparison.
Hornsey, V S; McColl, K; Drummond, O; Macgregor, I R; Prowse, C V
2008-08-01
New platelet storage systems, such as changes in the plastic of the storage bags, require validation. In this study, pooled buffy coat platelets stored in Fresenius/NPBI polyolefin bags were compared with those stored in Fresenius/NPBI butyryl-trihexyl citrate (BTHC) plasticized polyvinyl chloride (PVC). The CompoSelect thrombocyte polishing filter system (1000 mL polyolefin bag) and the CompoStop F730 system (1300 mL BTHC-PVC bag) were used to prepare paired, plasma-suspended, buffy coat platelet concentrates. Samples were taken up to day 7 for in vitro analysis. In a separate experiment, 12 units were prepared using the CompoStop F730 system and samples taken after leucofiltration for FXIIa assay. By day 7, platelet concentrates stored in BTHC-PVC demonstrated significantly higher pH levels (7.32 +/- 0.05 vs. 7.26 +/- 0.05) and a greater degree of cell lysis as shown by increased lactate dehydrogenase levels (497 +/- 107 vs. 392 +/- 81 U L(-1)). The supernatants contained higher concentrations of soluble P-selectin and the chemokine 'regulated on activation, normal T-cell expressed and presumably secreted', which are released from the alpha-granules during activation. The ATP concentrations were significantly lower in BTHC-PVC. Platelet counts, mean platelet volume and hypotonic shock response were similar for both bags. FXIIa antigen concentrations were 0.6 +/- 0.2 ng mL(-1) indicating that activation of the contact factor pathway had not occurred. Although the CompoStop F730 leucoreduction filter did not activate the contact system, platelets stored in 100% plasma in BTHC-PVC bags demonstrated different in vitro characteristics from those stored in polyolefin. Further work is required to demonstrate whether these differences will affect in vivo recovery and survival.
Sandgren, P; Hild, M; Sjödin, A; Gulliksson, H
2010-11-01
The novel TACSI system is designed for automated preparation of platelets (PLTs) from pooled buffy coats (BCs). One TACSI device will handle 6 units at the same time. The aim of our in vitro study is to investigate the effects of using this automated equipment with subsequent storage in two different plastic containers and to compare these results with PLTs prepared by the OrbiSac system. Buffy-coat-derived PLTs (n=8) were prepared by using the TACSI system, including storage in polyvinyl chloride (PVC)-based plastic containers with di, n-decyl phthalate (DnDP) (TACSI R) and BTHC (TACSI T)-based plasticizers. As a reference, the OrbiSac System was used to prepare PLTs (n=8) with subsequent storage in a PVC plastic container with a citrate-based plasticizer (BTHC). In total, 16 TACSI and eight reference units, supplied by approximately 30% plasma and 70% SSP+, were analysed for various in vitro variables during the 7-day storage period. No significant difference in PLT counts, LDH, mean platelet volume (MPV) and adenosine triphosphate between the groups was detected. Glucose was lower (P<0·05) and lactate was higher (P<0·05) in TACSI R vs. OrbiSac. With exception of day 7 (P<0·05 TACSI R vs. OrbiSac), HSR reactivity were not different between groups. Extent of shape change was lower and CD62P higher in TACSI T when compared with TACSI R and OrbiSac units (P<0·05). pH was maintained at >6·8 (day 7) and swirling remained at the highest level (score=2) for all units throughout storage. Platelets prepared by the TACSI system with subsequent storage in two different PVC-based plastic containers were equivalent to reference PLTs with regard to in vitro characteristics during 7 days of storage. © 2010 The Author(s). Vox Sanguinis © 2010 International Society of Blood Transfusion.
The role of microRNAs in platelet biology during storage.
Yan, Yuzhong; Zhang, Jingjun; Zhang, Qi; Chen, Yanping; Zhu, Xinfang; Xia, Rong
2017-04-01
Platelet storage lesions seriously affect the quality of stored platelets, even causing them to be ineffective in vivo after transfusion. Past research have been focused on what mechanism(s) cause the formation of storage lesions. One proposed mechanism is microRNAs (miRNAs)-based molecular regulation of the platelet mRNAs that are relevant to the storage lesion. Platelets continue to translate proteins from mRNA while in a storage environment. A strong correlation exists between the platelet transcriptome and its subsequent proteomic profile, which supports de novo platelet translational capabilities. Thus, miRNA may play a crucial role in platelet biology during storage conditions. Importantly, this suggests the exciting possibility of post-transcriptional regulation of gene expression in platelets that are in storage. Given this, the differential profiling of miRNAs could be a useful tool in identifying changes to ex vivo stored platelets. Any identified miRNAs could then be considered as potential markers to assess the viability of platelet concentrates. The present review summarizes the current experimental and clinical evidence that clarifies the role miRNAs play during platelet ex vivo storage. Copyright © 2016 Elsevier Ltd. All rights reserved.
Rogowska, Anna; Chabowska, Anna Małgorzata; Lipska, Alina; Boczkowska-Radziwon, Barbara; Bujno, Magdalena; Rusak, Tomasz; Dziemianczuk, Mateusz; Radziwon, Piotr
2016-05-01
In radiofrequency identification (RFID) systems used in labeling of blood components, blood cells are subjected to the direct influence of electromagnetic waves throughout the storage period. The aim of this study was to prove the safety of storage of platelet concentrates (PCs) in containers labeled with RFID tags. Ten pooled PCs obtained from 12 buffy coats each suspended in additive solution were divided into three separate containers that were assigned to three groups: control, PCs labeled with ultrahigh frequency (UHF) range tags and exposed to 915-MHz radio waves, and PCs labeled with high-frequency (HF) range tags and exposed to 13.56-MHz radio waves. PCs were stored at 20 to 24°C for 7 days. In vitro tests of platelet (PLT) function were performed on the first, fifth, and seventh days of storage. There were no significant differences in pH; hypotonic shock resistance; surface expression of CD62P, CD42a, or CD63; release of PLT-derived microparticles; PLT aggregation; and number of PLTs between PCs stored at a constant exposure to radio waves of two different frequencies and the control group on the first, fifth, and seventh days of storage. The results of the study indicate no impact of electromagnetic radiation generated in HF and UHF RFID systems and constant contact with the tags on the quality of stored PCs. © 2016 AABB.
Comparison of bacterial attachment to platelet bags with and without preconditioning with plasma.
Loza-Correa, M; Kalab, M; Yi, Q-L; Eltringham-Smith, L J; Sheffield, W P; Ramirez-Arcos, S
2017-07-01
Canadian Blood Services produces apheresis and buffy coat pooled platelet concentrates (PCs) stored in bags produced by two different manufacturers (A and B, respectively), both made of polyvinyl chloride-butyryl trihexyl citrate. This study was aimed at comparing Staphylococcus epidermidis adhesion to the inner surface of both bag types in the presence or absence of plasma factors. Sets (N = 2-6) of bags type A and B were left non-coated (control) or preconditioned with platelet-rich, platelet-poor or defibrinated plasma (PRP, PPP and DefibPPP, respectively). Each bag was inoculated with a 200-ml S. epidermidis culture adjusted to 0·5 colony-forming units/ml. Bags were incubated under platelet storage conditions for 7 days. After culture removal, bacteria attached to the plastic surface were either dislodged by sonication for bacterial quantification or examined in situ by scanning electron microscopy (SEM). Higher bacterial adhesion was observed to preconditioned PC bags than control containers for both bag types (P < 0·0001). Bacterial attachment to preconditioned bags was confirmed by SEM. Bacteria adhered equally to both types of containers in the presence of PRP, PPP and DefibPPP residues (P > 0·05). By contrast, a significant increase in bacterial adherence was observed to type A bags compared with type B bags in the absence of plasma (P < 0·05) [Correction added on 16 June 2017, after first online publication: this sentence has been corrected]. The ability of S. epidermidis to adhere to preconditioned platelet collection bags depends on the presence of plasma factors. Future efforts should be focused on reducing plasma proteins' attachment to platelet storage containers to decrease subsequent bacterial adhesion. © 2017 International Society of Blood Transfusion.
Exploratory studies of extended storage of apheresis platelets in a platelet additive solution (PAS)
Corson, Jill; Jones, Mary Kay; Christoffel, Todd; Pellham, Esther; Bailey, S. Lawrence; Bolgiano, Doug
2014-01-01
To evaluate the poststorage viability of apheresis platelets stored for up to 18 days in 80% platelet additive solution (PAS)/20% plasma, 117 healthy subjects donated platelets using the Haemonetics MCS+, COBE Spectra (Spectra), or Trima Accel (Trima) systems. Control platelets from the same subjects were compared with their stored test PAS platelets by radiolabeling their stored and control platelets with either 51chromium or 111indium. Trima platelets met Food and Drug Administration poststorage platelet viability criteria for only 7 days vs almost 13 days for Haemonetics platelets; ie, platelet recoveries after these storage times averaged 44 ± 3% vs 49 ± 3% and survivals were 5.4 ± 0.3 vs 4.6 ± 0.3 days, respectively. The differences in storage duration are likely related to both the collection system and the storage bag. The Spectra and Trima platelets were hyperconcentrated during collection, and PAS was added, whereas the Haemonetics platelets were elutriated with PAS, which may have resulted in less collection injury. When Spectra and Trima platelets were stored in Haemonetics’ bags, poststorage viability was significantly improved. Platelet viability is better maintained in vitro than in vivo, allowing substantial increases in platelet storage times. However, implementation will require resolution of potential bacterial overgrowth during storage. PMID:24258816
Pharmacologic Agents in the Management of Bleeding Disorders
1990-01-01
and the antifibrinolytic drugs tranexamic acid in preinfusion plasma,2 2 increase platelet adheience.23 (AMCA) and epsilon- aminocaproic acid (EACA...215-9. aminocaproic acid in the treatment of patients with acute pro- 54. Schulman S, Johnsson H, Egberg N, Blomback M. DDAVP- myclocytie leukemia and...shortens the bleeding time in storage pool 79. Gardner Fli, Helmer RE Ill. Aminocaproic acid . Use in control deficiency. Ann Intern Med 1988;108:65-7. of
EXTENDED STORAGE OF BUFFY-COAT PLATELET CONCENTRATES IN PLASMA OR A PLATELET ADDITIVE SOLUTION
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
Carvalho, Helena; Alguero, Carmen; Santos, Matilde; de Sousa, Gracinda; Trindade, Helder; Seghatchian, Jerard
2006-04-01
Platelets are known to undergo shape change, activation, a release reaction and apoptosis/necrosis during processing and storage, all of which are collectively known as the platelet storage lesion. Any additional processing may have some deleterious impact on platelet activability and functional integrity, which need to be investigated. This preliminary investigation was undertaken to establish the combined effects of standard platelet storage media and the intercept pathogen reduction technology on platelet activation and activability during 7 day storage, using buffy-coat derived platelets in standard storage media containing 35% plasma (N=24). P-selectin (CD62p) expression, a classical marker of platelet activation, and phosphatidylserine (PS) exposure on the platelet surface membrane, a hallmark of cellular necrosis/apoptosis, were both measured by flow cytometry. The results reveal significant increases in activation, from an average of 22.7% on day 1 before treatment to 31.6% on day 2 after treatment and 58.7% at the end of storage. Concomitantly, the basal expression of PS was slightly increased from 1.9% to 2.8% at day 2 after treatment and 7.3% at the end of storage. However, the functional reserve of platelets during storage, which reflects their capability to undergo activation and the release reaction when platelets were challenged with either calcium ionophore or thrombin, was relatively well maintained. These preliminary data confirm the earlier data on the use of intercept, and for the first time, based on the assessment of platelet functional integrity, suggest that platelet functional reserve is relatively well maintained, with little change in the formation of apoptotic cells.
Waters, L; Cameron, M; Padula, M P; Marks, D C; Johnson, L
2018-05-01
Conventional storage of platelet concentrates limits their shelf life to between 5 and 7 days due to the risk of bacterial proliferation and the development of the platelet storage lesion. Cold storage and cryopreservation of platelets may facilitate extension of the shelf life to weeks and years, and may also provide the benefit of being more haemostatically effective than conventionally stored platelets. Further, treatment of platelet concentrates with pathogen inactivation systems reduces bacterial contamination and provides a safeguard against the risk of emerging and re-emerging pathogens. While each of these alternative storage techniques is gaining traction individually, little work has been done to examine the effect of combining treatments in an effort to further improve product safety and minimize wastage. This review aims to discuss the benefits of alternative storage techniques and how they may be combined to alleviate the problems associated with conventional platelet storage. © 2018 International Society of Blood Transfusion.
Singh, Sukhi; Shams Hakimi, Caroline; Jeppsson, Anders; Hesse, Camilla
2017-12-01
Platelet storage lesion is characterized by morphological changes and impaired platelet function. The collection method and storage medium may influence the magnitude of the storage lesion. The aim of this study was to compare the newly introduced interim platelet unit (IPU) platelet concentrates (PCs) (additive solution SSP+, 40% residual plasma content) with the more established buffy-coat PCs (SSP, 20% residual plasma content) and apheresis PCs (autologous plasma) in terms of platelet storage lesions. Thirty PCs (n=10 for each type) were assessed by measuring metabolic parameters (lactate, glucose, and pH), platelet activation markers, and in vitro platelet aggregability on days 1, 4, and 7 after donation. The expression of platelet activation markers CD62p (P-selectin), CD63 (LAMP-3), and phosphatidylserine was measured using flow cytometry and in vitro aggregability was measured with multiple electrode aggregometry. Higher platelet activation and lower in vitro aggregability was observed in IPU than in buffy-coat PCs on day 1 after donation. In contrast, metabolic parameters, expression of platelet activation markers, and in vitro aggregability were better maintained in IPU than in buffy-coat PCs at the end of the storage period. Compared to apheresis PCs, IPU PCs had higher expression of activation markers and lower in vitro aggregability throughout storage. In conclusion, the results indicate that there are significant differences in platelet storage lesions between IPU, buffy-coat, and apheresis PCs. The quality of IPU PCs appears to be at least comparable to buffy-coat preparations. Further studies are required to distinguish the effect of the preparation methods from storage conditions. Copyright © 2017 Elsevier Ltd. All rights reserved.
Platelet concentrates for transfusion-metabolic and storage aspects.
Farrugia, A
1994-01-01
Transfusion of platelets concentrated from donated blood is an established therapeutic modality in clinical medicine. Over the past 25 years much effort has gone into optimising the conditions for the collection, preparation and storage of platelets for transfusion. Despite significant advances, platelet production is still a costly process requiring a dedicated environment and the use of specially formulated plastic storage containers. A progressive lesion over storage limits the shelf life and the availability of donated platelets, while the need to store platelets in the donor's autologous plasma also results in a loss of valuable fresh plasma for fractionation. Recent studies have addressed the issues of platelet quality and plasma economy by examining the possibility of storing platelets in a synthetic medium. Platelets stored in a variety of crystalloid solutions have been shown to retain in vitro and in vivo properties equivalent or superior to platelets stored in autologous donor plasma. Some additional insight has been gained on the metabolic patterns of stored platelets. In particular, studies have shown that, under these conditions, platelets are unable to oxidise dextrose to any significant extent, and that dextrose is invariably broken down to lactate, irrespective of the oxygen tensions in the platelet's environment. This in turn leads to the metabolic lesion of platelet storage, whereby low pH results in loss of platelet viability. Platelets stored in synthetic dextrose-free media are capable of maintaining aerobic ATP generation, and acetate-a component of many media studied-has been shown to be metabolised by platelets. Similarly, platelets prepared from blood collected into a dextrose-free anticoagulant have satisfactory properties both when suspended in autologous plasma or in a dextrose-free synthetic medium. The requirements for storage in special, high gas-permeable, containers, and for constant agitation during storage, were both found to be unnecessary when dextrose was excluded from the platelet's environment. These developments suggest that manipulation of the platelet's metabolic pattern during blood bank storage may allow significant benefits in plasma economy as well as in decreasing the cost of platelet delivery to patients.
Platelet Storage Lesions: What More Do We Know Now?
Ng, Monica Suet Ying; Tung, John-Paul; Fraser, John Francis
2018-04-17
Platelet concentrate (PC) transfusions are a lifesaving adjunct to control and prevent bleeding in cancer, hematologic, surgical, and trauma patients. Platelet concentrate availability and safety are limited by the development of platelet storage lesions (PSLs) and risk of bacterial contamination. Platelet storage lesions are a series of biochemical, structural, and functional changes that occur from blood collection to transfusion. Understanding of PSLs is key for devising interventions that prolong PC shelf life to improve PC access and wastage. This article will review advancements in clinical and mechanistic PSL research. In brief, exposure to artificial surfaces and high centrifugation forces during PC preparation initiate PSLs by causing platelet activation, fragmentation, and biochemical release. During room temperature storage, enhanced glycolysis and reduced mitochondrial function lead to glucose depletion, lactate accumulation, and product acidification. Impaired adenosine triphosphate generation reduces platelet capacity to perform energetically demanding processes such as hypotonic stress responses and activation/aggregation. Storage-induced alterations in platelet surface proteins such as thrombin receptors and glycoproteins decrease platelet aggregation. During storage, there is an accumulation of immunoactive proteins such as leukocyte-derive cytokines (tumor necrosis factor α, interleukin (IL) 1α, IL-6, IL-8) and soluble CD40 ligand which can participate in transfusion-related acute lung injury and nonhemolytic transfusion reactions. Storage-induced microparticles have been linked to enhanced platelet aggregation and immune system modulation. Clinically, stored PCs have been correlated with reduced corrected count increment, posttransfusion platelet recovery, and survival across multiple meta-analyses. Fresh PC transfusions have been associated with superior platelet function in vivo; however, these differences were abrogated after a period of circulation. There is currently insufficient evidence to discern the effect of PSLs on transfusion safety. Various bag and storage media changes have been proposed to reduce glycolysis and platelet activation during room temperature storage. Moreover, cryopreservation and cold storage have been proposed as potential methods to prolong PC shelf life by reducing platelet metabolism and bacterial proliferation. However, further work is required to elucidate and manage the PSLs specific to these storage protocols before its implementation in blood banks. Copyright © 2018 Elsevier Inc. All rights reserved.
Sorokin, Victoria; Alkhoury, Razan; Al-Rawabdeh, Sura; Houston, Ronald H; Thornton, David; Kerlin, Bryce; O'Brien, Sarah; Baker, Peter; Boesel, Carl; Uddin, Minhaj; Yin, Han; Kahwash, Samir
This study sought to determine delta granule normal ranges for children and to validate methodology for the appropriate diagnosis of delta granule deficiency (storage pool disease) by using the whole-mount technique in electron microscopy. Specimens obtained from 40 healthy volunteers (2 months of age through 21 years old, 21 females and 19 males) were tested. Results showed dense granules/platelet (DG/Plt) ranged from 1.78 to 5.25. The 5th percentile was 1.96 DG/Plt with an overall mean ± SEM 3.07 ± 0.12 DG/Plt. In comparison, a previously published lower cutoff value, 3.68 DG/Plt, was significantly higher than the mean from our volunteers (P < 0.0001). We found no variability in dense granules/platelet based on race or sex and no significant variation by age subgroup. Pending wider studies, the value of 2 DG/Plt is a more appropriate lower limit of normal. In the absence of wider studies (in healthy volunteers and patients), laboratories should consider establishing their own reference ranges.
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 platelet...
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 platelet...
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 platelet...
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 platelet...
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...
Loh, Y S; Dean, M M; Johnson, L; Marks, D C
2015-11-01
Pathogen inactivation (PI) and storage may alter the immunomodulatory capacity of platelets (PLTs). The aim of this study was to examine the effect of PI (Riboflavin and ultraviolet light treatment) and storage on the capacity of PLTs to induce cytokine responses in recipient inflammatory cells. A pool and split design was used to prepare untreated and PI-treated buffy coat-derived platelet concentrates (PCs). Samples were taken on days 2 and 7 postcollection and incubated with ABO/RhD-matched fresh whole blood for 6 h with or without lipopolysaccharide (LPS). The intracellular production of IP-10, MCP-1, MIP-1α, IL-8, IL-6, IL-10, IL-12, TNF-α and MIP-1β in monocytes and neutrophils was assessed using flow cytometry. Complement proteins in PLT supernatants were measured using a cytometric bead array. PLTs and PLT supernatant (both untreated and PI-treated) resulted in modulation of intracellular MIP-1β and IL-12 production in monocytes. Compared to untreated PLTs, PI-treated PLTs resulted in significantly lower LPS-induced monocyte IL-12 production (day 7). The concentration of C3a and C5a (and their desArg forms) was significantly increased in PLT supernatants following PI. PI results in decreased LPS-induced monocyte IL-12 production and increased complement activation. The association between platelet-induced complement activation and IL-12 production warrants further investigation. © 2015 International Society of Blood Transfusion.
Ghasemzadeh, Mehran; Hosseini, Ehteramolsadat; Roudsari, Zahra Oushyani; Zadkhak, Parvin
2018-03-01
Platelets storage lesion is mainly orchestrated by platelet activating signals during storage. Reactive oxygen species (ROS) are being considered as important signaling molecules modulating platelet function while their production has also been shown to be augmented by platelet activation. This study investigated to what extent endogenous ROS generation during platelet storage could be correlated with platelet receptor shedding, microvesiculation and adhesive function. 10 PRP-platelet concentrates were subjected to flow cytometry analysis to examine the generation of intraplatelet ROS on days 1, 5 and 7 after storage. In 5 day-stored platelets considering 40% of ROS generation as a cutoff point, samples were divided into two groups of those with higher or lower levels of ROS. The expression of adhesion receptors (GPVI, GPIbα), the amount of microparticles and phosphatidylserine exposure in each group were then examined by flow cytometry. Platelet receptor shedding and adhesion to collagen matrix were respectively measured by western blotting and microscopic assays. Our data showed lowered expression of GPIbα (p < 0.05) and GPVI in samples with ROS > 40% than those with ROS ≤ 40%, whereas receptors shedding and microvesiculation were (p < 0.05) elevated in platelets with higher levels of ROS. Functionally, we observed significantly (p < 0.05) lower levels of platelet adhesion to collagen matrix in samples with ROS generation more than 40%. Taken together, we showed correlations between intraplatelet ROS generation and either platelet receptors or microparticle shedding as well as platelet adhesive capacity to collagen. These findings suggest that augmented ROS generation during storage might be relevant to down-regulation of platelet adhesive function. Copyright © 2018 Elsevier Ltd. All rights reserved.
Södergren, A L; Tynngård, N; Berlin, G; Ramström, S
2016-02-01
Storage lesions may prevent transfused platelets to respond to agonists and arrest bleeding. The aim of this study was to evaluate and quantify the capacity of platelet activation during storage using flow cytometry and new markers of platelet activation. Activation responses of platelets prepared by apheresis were measured on days 1, 5, 7 and 12. In addition, comparisons were made for platelet concentrates stored until swirling was affected. Lysosome-associated membrane protein-1 (LAMP-1), P-selectin and phosphatidylserine (PS) exposure were assessed by flow cytometry on platelets in different subpopulations in resting state or following stimulation with platelet agonists (cross-linked collagen-related peptide (CRP-XL), PAR1- and PAR4-activating peptides). The ability to form subpopulations upon activation was significantly decreased already at day 5 for some agonist combinations. The agonist-induced exposure of PS and LAMP-1 also gradually decreased with time. Spontaneous exposure of P-selectin and PS increased with time, while spontaneous LAMP-1 exposure was unchanged. In addition, agonist-induced LAMP-1 expression clearly discriminated platelet concentrates with reduced swirling from those with retained swirling. This suggests that LAMP-1 could be a good marker to capture changes in activation capacity in stored platelets. The platelet activation potential seen as LAMP-1 exposure and fragmentation into platelet subpopulations is potential sensitive markers for the platelet storage lesion. © 2015 International Society of Blood Transfusion.
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.
The Slc35d3 gene, encoding an orphan nucleotide sugar transporter, regulates platelet-dense granules
Chintala, Sreenivasulu; Tan, Jian; Gautam, Rashi; Rusiniak, Michael E.; Guo, Xiaoli; Li, Wei; Gahl, William A.; Huizing, Marjan; Spritz, Richard A.; Hutton, Saunie; Novak, Edward K.; Swank, Richard T.
2007-01-01
Platelet dense granules are lysosome-related organelles which contain high concentrations of several biologically important low-molecular-weight molecules. These include calcium, serotonin, adenine nucleotides, pyrophosphate, and polyphosphate, which are necessary for normal blood hemostasis. The synthesis of dense granules and other lysosome-related organelles is defective in inherited diseases such as Hermansky-Pudlak syndrome (HPS) and Chediak-Higashi syndrome (CHS). HPS and CHS mutations in 8 human and at least 16 murine genes have been identified. Previous studies produced contradictory findings for the function of the murine ashen (Rab27a) gene in platelet-dense granules. We have used a positional cloning approach with one line of ashen mutants to establish that a new mutation in a second gene, Slc35d3, on mouse chromosome 10 is the basis of this discrepancy. The platelet-dense granule defect is rescued in BAC transgenic mice containing the normal Slc35d3 gene. Thus, Slc35d3, an orphan member of a nucleotide sugar transporter family, specifically regulates the contents of platelet-dense granules. Unlike HPS or CHS genes, it has no apparent effect on other lysosome-related organelles such as melanosomes or lysosomes. The ash-Roswell mouse mutant is an appropriate model for human congenital-isolated delta-storage pool deficiency. PMID:17062724
Wagner, S J; Skripchenko, A; Seetharaman, S; Kurtz, J
2015-04-01
Previous studies with p38MAPK inhibitors at room temperature demonstrated that they improve a large number of platelet storage parameters, but cannot substantially inhibit p38MAPK activation nor protect against widespread decrements in platelet quality parameters during 4 °C storage. In this study, platelet quality parameters and inhibition of p38MAPK by VX-702 were studied after incubation of platelets at 16 °C without agitation, suboptimal storage conditions which produce moderate platelet decrements. Trima apheresis units were collected and aliquoted into three 60-ml CLX storage bags: (i) a control aliquot which was held at 20-24 °C with constant agitation; (ii) a test aliquot which was held at 20-24 °C with agitation until Day 2, when it was reincubated at 16 ± 1 °C for 24 ± 0·5 h without agitation and then returned 20-24 °C with agitation; (iii) a test aliquot containing 1 μm VX-702 stored in an identical fashion as aliquot 2. Aliquots were tested for an array of platelet storage parameters and p38MAPK activation on Days 1, 4 and 7. Many platelet storage parameters and p38MAPK activation were adversely affected by 24-h incubation at 16 °C without agitation. With the exception of ESC, addition of VX-702 prevented p38MAPK activation and the decrements in most observed parameters. Unlike 4 °C storage, VX-702 prevents activation of p38MAPK and decrements in many platelet storage parameters after exposure to 16 °C without agitation for 24 h. © 2014 International Society of Blood Transfusion.
Thiele, Thomas; Iuga, Cristina; Janetzky, Susann; Schwertz, Hansjorg; Gesell Salazar, Manuela; Fürll, Birgit; Völker, Uwe; Greinacher, Andreas; Steil, Leif
2012-12-05
Production and storage of platelet concentrates (PC) induce protein changes in platelets leading to impaired platelet function. This study aimed to identify signaling pathways involved in the development of early platelet storage lesions in apheresis-PCs stored in plasma or additive solution (PAS). Apheresis-PCs from four donors were stored in plasma or in PAS at 22°C (n=4 each). Platelets were analyzed at day 0 (production day) and after 1, 6 and 9 days of storage. Platelet response to agonists (TRAP, collagen, ADP) and to hypotonic shock decreased, CD62P expression increased in both storage media over time. Using DIGE 1550 protein spots were monitored and compared to baseline values at day 0. Platelets in plasma displayed changes in 352 spots (166/day 1, 263/day 6 and 201/day 9); in PAS 325 spots changed (202/day 1, 221/day 6, 200/day 9). LC-ESI-MS/MS analysis of 405 platelet proteins revealed 32 proteins changed during storage in plasma (9/day 1, 15/day 6 and 26/day 9) and 28 in PAS (5/day 1, 20/day 6, 26/day 9). Ingenuity pathway analysis found integrin-αII(b)β(3) and focal adhesion signaling pathways involved in early alterations, being confirmed by Western blotting. Corresponding mRNAs in platelets were identified by next generation sequencing for 84 changed proteins. Integrin-αII(b)β(3) and focal adhesion signaling cause irreversible early storage lesions in apheresis platelets. This article is part of a Special Issue entitled: Integrated omics. Copyright © 2012 Elsevier B.V. All rights reserved.
Marcoux, Geneviève; Duchez, Anne-Claire; Rousseau, Matthieu; Lévesque, Tania; Boudreau, Luc H; Thibault, Louis; Boilard, Eric
2017-05-01
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.
Overview of platelet physiology and laboratory evaluation of platelet function.
Rodgers, G M
1999-06-01
Appropriate laboratory testing for the platelet-type bleeding disorders hinges on an adequate assessment in the history and physical examination. Patients with histories and screening laboratory results consistent with coagulation disorders (hemophilia, disseminated intravascular coagulation) are not appropriate candidates for platelet function testing. In contrast, patients with a lifelong history of platelet-type bleeding symptoms and perhaps a positive family history of bleeding would be appropriate for testing. Figure 6 depicts one strategy to evaluate these patients. Platelet morphology can easily be evaluated to screen for two uncommon qualitative platelet disorders: Bernard-Soulier syndrome (associated with giant platelets) and gray platelet syndrome, a subtype of storage pool disorder in which platelet granulation is morphologically abnormal by light microscopy. If the bleeding disorder occurred later in life (no bleeding with surgery or trauma early in life), the focus should be on acquired disorders of platelet function. For those patients thought to have an inherited disorder, testing for vWD should be done initially because approximately 1% of the population has vWD. The complete vWD panel (factor VIII coagulant activity, vWf antigen, ristocetin cofactor activity) should be performed because many patients will have abnormalities of only one particular panel component. Patients diagnosed with vWD should be classified using multimeric analysis to identify the type 1 vWD patients likely to respond to DDAVP. If vWD studies are normal, platelet aggregation testing should be performed, ensuring that no antiplatelet medications have been ingested at least 1 week before testing. If platelet aggregation tests are normal and if suspicion for an inherited disorder remains high, vWD testing should be repeated. The evaluation of thrombocytopenia may require bone marrow examination to exclude primary hematologic disorders. If future studies with thrombopoietin assays confirm preliminary results, however, the bone marrow examination of certain patients may be replaced by a thrombopoietin level.
Heuft, H-G; Goudeva, L; Krauter, J; Peest, D; Buchholz, S; Tiede, A
2013-07-01
To evaluate the clinical effect of platelet concentrate (PC) transfusions after PC storage time reduction to 4 days. This was a single-centre cohort study comparing two 3-month periods of time, before and after the reduction of PC storage time from 5 to 4 days. Seventy-seven consecutive patients with PC transfusions were enrolled after blood stem cell transplantation. Corrected platelet count increment (CCI) on the morning after transfusion, time to next platelet transfusion, need for red blood cell (RBC) transfusion and clinical bleeding symptoms were compared. Platelet concentrate storage time was reduced between period 1 (storage for up to 5 days, median storage time 78 h, range 11-136 h) and period 2 (storage for up to 4 days, median storage time 53 h, range 11-112 h). Patients were comparable for age, weight, body surface area, underlying disorder, type of transplantation and transfused platelet dose. The CCI increased from a median of 4 (range 0-20) to 8 (0-68) × 10(9) /l per 10(11) platelets/m(2) (P < 0·0001). Time to next PC transfusion increased from 1·1 to 2·0 days (P < 0·0001). Any bleeding symptom was noted in 20 of 36 patients (56%) vs. 9/41 patients (22%, P < 0·01). Nose bleeds, haematuria and bleeding at more than one site were significantly reduced. Frequency of RBC transfusion within 5 days after PC transfusion was reduced from 74 to 58% (P < 0·0001). Platelet concentrate storage time shortening was associated with highly significant CCI increase, reduced RC needs and lower patient numbers with bleeding events. © 2013 International Society of Blood Transfusion.
Platelet utilization: a Canadian Blood Services research and development symposium.
Webert, Kathryn E; Alam, Asim Q; Chargé, Sophie B; Sheffield, William P
2014-04-01
Considerable progress has been made in recent years in understanding platelet biology and in strengthening the clinical evidence base around platelet transfusion thresholds and appropriate platelet dosing. Platelet alloimmunization rates have also declined. Nevertheless, controversies and uncertainties remain that are relevant to how these products can best be used for the benefit of platelet transfusion recipients. Platelets are unique among the blood products directly derived from whole blood or apheresis donations in requiring storage, with shaking, at ambient temperature. Storage is accordingly constrained between the need to limit the growth of any microbes in the product and the need to minimize losses in platelet function associated with storage. Proteomic and genomic approaches are being applied to the platelet storage lesion. Platelet inventory management is made challenging by these constraints. Although bacterial screening has enhanced the safety of platelet transfusions, pathogen reduction technology may offer further benefits. Continuing clinical investigations are warranted to understand the value of transfusing platelets prophylactically or only in response to bleeding in different patient groups and how best to manage the most grievously injured trauma patients. Patients refractory to platelet transfusions also require expert clinical management. The engineering of platelet substitute products is an active area of research, but considerable hurdles remain before any clinical uses may be contemplated. Roles for platelets in biological areas distinct from hemostasis are also emerging. Platelet utilization is variably affected by all of the above factors, by demographic changes, by new medications, and by new patient care approaches. Copyright © 2014 Elsevier Inc. All rights reserved.
Platelet von Willebrand factor in Hermansky-Pudlak syndrome.
McKeown, L P; Hansmann, K E; Wilson, O; Gahl, W; Gralnick, H R; Rosenfeld, K E; Rosenfeld, S J; Horne, M K; Rick, M E
1998-10-01
The Hermansky-Pudlak Syndrome (HPS) is an autosomal recessive inherited disorder characterized by oculocutaneous albinism, tissue accumulation of ceroid pigment, and a mild to moderate bleeding diathesis attributed to storage-pool deficient (SPD) platlets. Patients have platelet aggregation and release abnormalities. In addition, low levels of plasma von Willebrand factor (vWF) antigen in some HPS patients have been associated with a greater bleeding tendency than would be predicted from either condition alone. Other HPS patients have severe bleeding despite normal levels of plasma vWF, suggesting that at least one additional factor is responsible for their bleeding diathesis. Because platelet vWF levels have been well correlated with clinical bleeding times in patients with von Willebrand's disease, we have measured the platelet vWF activity and antigen levels in 30 HPS patients and have attempted to correlate their clinical bleeding with these values. The platelet vWF activity levels in patients was significantly lower than that of normal subjects (P < 0.0001). The patients as a group also had slightly lower values of plasma vWF activity when compared with normals (P-0.03). In 11 of the HPS patients, the multimeric structure of plasma vWF showed a decrease in the high molecular weight multimers and an increase in the low molecular weight multimers. In correlating the platelet and plasma vWF values with the bleeding histories, we were not able to show a predictable relationship in the majority of the patients.
Scott Murphy, MD: platelet storage pioneer.
Vassallo, Ralph
2011-04-01
Dr Scott Murphy was a leading authority in the science of platelet storage for transfusion. Among the first to advocate room-temperature storage, his work enabled the extension of platelet shelf life, which facilitated the development of aggressive medical and surgical therapies requiring intensive platelet support. During a 38-year career, he characterized the metabolism of stored platelets, worked to optimize storage conditions, and developed a landmark standard for the assessment of platelet quality. As a clinical hematologist, he was devoted to his patients, providing compassionate, comforting care. He was also a cherished mentor, a sage advisor, a fast friend, and a dedicated family man. Dr Murphy was one of those rare, great, humble men who inspire and enrich everyone around him. Truly one of transfusion medicine's leaders, Scott is remembered by those whose lives he touched for his warmth, wit, and wisdom. Copyright © 2011 Elsevier Inc. All rights reserved.
Wood, Ben; Padula, Matthew P; Marks, Denese C; Johnson, Lacey
2016-10-01
Platelets (PLTs) are currently stored at room temperature (22°C), which limits their shelf life, primarily due to the risk of bacterial growth. Alternatives to room temperature storage include PLT refrigeration (2-6°C), which inhibits bacterial growth, thus potentially allowing an extension of shelf life. Additionally, refrigerated PLTs appear more hemostatically active than conventional PLTs, which may be beneficial in certain clinical situations. However, the mechanisms responsible for this hemostatic function are not well characterized. The aim of this study was to assess the protein profile of refrigerated PLTs in an effort to understand these functional consequences. Buffy coat PLTs were pooled, split, and stored either at room temperature (20-24°C) or under refrigerated (2-6°C) conditions (n = 8 in each group). PLTs were assessed for changes in external receptor expression and actin filamentation using flow cytometry. Intracellular proteomic changes were assessed using two-dimensional gel electrophoresis and Western blotting. PLT refrigeration significantly reduced the abundance of glycoproteins (GPIb, GPIX, GPIIb, and GPIV) on the external membrane. However, refrigeration resulted in the increased expression of high-affinity integrins (αIIbβ3 and β1) and activation and apoptosis markers (CD62P, CD63, and phosphatidylserine). PLT refrigeration substantially altered the abundance and localization of several cytoskeletal proteins and resulted in an increase in actin filamentation, as measured by phalloidin staining. Refrigerated storage of PLTs induces significant changes in the expression and localization of both surface-expressed and intracellular proteins. Understanding these proteomic changes may help to identify the mechanisms resulting in the refrigeration-associated alterations in PLT function and clearance. © 2016 AABB.
Handigund, Mallikarjun; Bae, Tae Won; Lee, Jaehyeon; Cho, Yong Gon
2016-02-01
Platelets play a vital role in hemostasis and thrombosis, and their demand and usage has multiplied many folds over the years. However, due to the short life span and storage constraints on platelets, it is allowed to store them for up to 7 days at room temperature (RT); thus, there is a need for an alternative storage strategy for extension of shelf life. Current investigation involves the addition of 50 mM N acetylcysteine (NAC) in refrigerated concentrates. Investigation results revealed that addition of NAC to refrigerated concentrates prevented platelet activation and reduced the sialidase activity upon rewarming as well as on prolonged storage. Refrigerated concentrates with 50 mM NAC expressed a 23.91 ± 6.23% of CD62P (P-Selectin) and 22.33 ± 3.42% of phosphotidylserine (PS), whereas RT-stored platelets showed a 46.87 ± 5.23% of CD62P and 25.9 ± 6.48% of phosphotidylserine (PS) after 5 days of storage. Further, key metabolic parameters such as glucose and lactate accumulation indicated reduced metabolic activity. Taken together, investigation and observations indicate that addition of NAC potentially protects refrigerated concentrates by preventing platelet activation, stabilizing sialidase activity, and further reducing the metabolic activity. Hence, we believe that NAC can be a good candidate for an additive solution to retain platelet characteristics during cold storage and may pave the way for extension of storage shelf life. Copyright © 2016 Elsevier Ltd. All rights reserved.
Beshkar, Pezhman; Hosseini, Ehteramolsadat; Ghasemzadeh, Mehran
2018-02-01
Regardless of different sources, methods or devices which are applied for preparation of therapeutic platelets, these products are generally isolated from whole blood by the sedimentation techniques which are based on PRP or buffy coat (BC) separation. As a general fact, platelet preparation and storage are also associated with some deleterious changes that known as platelet storage lesion (PSL). Although these alternations in platelet functional activity are aggravated during storage, whether technical issues within preparation can affect integrin activation and platelet adhesion to fibrinogen were investigated in this study. PRP- and BC-platelet concentrates (PCs) were subjected to flowcytometry analysis to examine the expression of platelet activation marker, P-selectin as well as active confirmation of the GPIIb/IIIa (α IIb β 3 ) on day 0, 1, 3 and 5 post-storage. Platelet adhesion to fibrinogen matrix was evaluated by fluorescence microscopy. Glucose concentration and LDH activity were also measured by colorimetric methods. The increasing P-selectin expression during storage was in a reverse correlation with PAC-1 binding (r = -0.67; p = .001). PRP-PCs showed the higher level of P-selectin expression than BC-PCs, whereas the levels of PAC-1 binding and platelet adhesion to fibrinogen matrix were significantly lower in PRP-PCs. Higher levels of active confirmation of the GPIIb/IIIa in BC-PCs were also associated with greater concentration of glucose in these products. We demonstrated the superior capacities of integrin activation and adhesion to fibrinogen for BC-PCs compared to those of PRP-PCs. These findings may provide more advantages for BC method of platelet preparation. Copyright © 2017 Elsevier Ltd. All rights reserved.
Schubert, Peter; Devine, Dana V
2010-01-03
Proteomics has brought new perspectives to the fields of hematology and transfusion medicine in the last decade. The steady improvement of proteomic technology is propelling novel discoveries of molecular mechanisms by studying protein expression, post-translational modifications and protein interactions. This review article focuses on the application of proteomics to the identification of molecular mechanisms leading to the deterioration of blood platelets during storage - a critical aspect in the provision of platelet transfusion products. Several proteomic approaches have been employed to analyse changes in the platelet protein profile during storage and the obtained data now need to be translated into platelet biochemistry in order to connect the results to platelet function. Targeted biochemical applications then allow the identification of points for intervention in signal transduction pathways. Once validated and placed in a transfusion context, these data will provide further understanding of the underlying molecular mechanisms leading to platelet storage lesion. Future aspects of proteomics in blood banking will aim to make use of protein markers identified for platelet storage lesion development to monitor proteome changes when alterations such as the use of additive solutions or pathogen reduction strategies are put in place in order to improve platelet quality for patients. (c) 2009 Elsevier B.V. All rights reserved.
Hexamoll DINCH plasticised PVC containers for the storage of platelets
Nair, C. S. Bhaskaran; Vidya, R.; Ashalatha, P. M.
2011-01-01
Introduction: Containers for the storage of platelets are made using polyvinyl chloride plasticised with di, (2-ethyl hexyl) phthalate, n-butyryl, tri (n-hexyl) citrate and tri (2-ethyl hexyl) mellitate or using special poly olefins without plasticiser. Of these, the first two have disadvantages such as plasticiser leaching and impairment of platelet function. Polyolefin bags cannot be HF welded or steam sterilized. Mellitate plasticised bags can store platelets well for five days but they are not completely phthalate free. Research and Development: We have developed a new generation of containers made of PVC plasticised with the non DEHP, non aromatic plasticiser,1,2- Cyclohexanedicarboxylic acid, diisononyl ester (Hexamoll DINCH) which can store platelets without loss of function for at least six days. Observation: The present studies show that DINCH plasticised PVC bags (TPL-167) are well suited for the storage of platelet concentrates for more than five days. Conclusion: The present studies show that the PVC plasticised with the non phthalate, non aromatic, non toxic plasticiser DINCH is a viable alternative to other existing containers for the storage of platelets for more than five days. PMID:21572709
Amorini, Angela M.; Tuttobene, Michele; Tomasello, Flora M.; Biazzo, Filomena; Gullotta, Stefano; De Pinto, Vito; Lazzarino, Giuseppe; Tavazzi, Barbara
2013-01-01
Background It is essential that the quality of platelet metabolism and function remains high during storage in order to ensure the clinical effectiveness of a platelet transfusion. New storage conditions and additives are constantly evaluated in order to achieve this. Using glucose as a substrate is controversial because of its potential connection with increased lactate production and decreased pH, both parameters triggering the platelet lesion during storage. Materials and methods In this study, we analysed the morphological status and metabolic profile of platelets stored for various periods in autologous plasma enriched with increasing glucose concentrations (13.75, 27.5 and 55 mM). After 0, 2, 4, 6 and 8 days, high energy phosphates (ATP, GTP, ADP, AMP), oxypurines (hypoxanthine, xanthine, uric acid), lactate, pH, mitochondrial function, cell lysis and morphology, were evaluated. Results The data showed a significant dose-dependent improvement of the different parameters in platelets stored with increasing glucose, compared to what detected in controls. Interestingly, this phenomenon was more marked at the highest level of glucose tested and in the period of time generally used for platelet transfusion (0–6 days). Conclusion These results indicate that the addition of glucose during platelet storage ameliorates, in a dose-dependent manner, the biochemical parameters related to energy metabolism and mitochondrial function. Since there was no correspondence between glucose addition, lactate increase and pH decrease in our experiments, it is conceivable that platelet derangement during storage is not directly caused by glucose through an increase of anaerobic glycolysis, but rather to a loss of mitochondrial functions caused by reduced substrate availability. PMID:22682337
Overview on platelet preservation: better controls over storage lesion.
Ohto, Hitoshi; Nollet, Kenneth E
2011-06-01
Platelet storage lesion (PSL), correlating with reduced in vivo recovery/survival and hemostatic capacity after transfusion, is characterized essentially by morphological and molecular evidence of platelet activation and energy consumption in the medium. Processes that limit shelf-life are multifactorial, and include both necrosis and apoptosis. PSL is greatly influenced by factors including duration of storage, temperature, ratio of platelet number to media volume, solution composition with respect to energy content and buffering capacity, and gas permeability of the container. Recent progress for slowing PSL has been made with storage media that more effectively fuel ATP production and buffer the inevitable effects of metabolism. Improved oxygen-permeability of containers also helps to maintain aerobic-dominant glycolysis. Patients stand to benefit from platelet products of higher intrinsic quality that store well until the moment of transfusion. Copyright © 2011. Published by Elsevier Ltd.
Salunkhe, Vishal; De Cuyper, Iris M; Papadopoulos, Petros; van der Meer, Pieter F; Daal, Brunette B; Villa-Fajardo, María; de Korte, Dirk; van den Berg, Timo K; Gutiérrez, Laura
2018-03-19
Platelet concentrates (PCs) represent a blood transfusion product with a major concern for safety as their storage temperature (20-24°C) allows bacterial growth, and their maximum storage time period (less than a week) precludes complete microbiological testing. Pathogen inactivation technologies (PITs) provide an additional layer of safety to the blood transfusion products from known and unknown pathogens such as bacteria, viruses, and parasites. In this context, PITs, such as Mirasol Pathogen Reduction Technology (PRT), have been developed and are implemented in many countries. However, several studies have shown in vitro that Mirasol PRT induces a certain level of platelet shape change, hyperactivation, basal degranulation, and increased oxidative damage during storage. It has been suggested that Mirasol PRT might accelerate what has been described as the platelet storage lesion (PSL), but supportive molecular signatures have not been obtained. We aimed at dissecting the influence of both variables, that is, Mirasol PRT and storage time, at the proteome level. We present comprehensive proteomics data analysis of Control PCs and PCs treated with Mirasol PRT at storage days 1, 2, 6, and 8. Our workflow was set to perform proteomics analysis using a gel-free and label-free quantification (LFQ) approach. Semi-quantification was based on LFQ signal intensities of identified proteins using MaxQuant/Perseus software platform. Data are available via ProteomeXchange with identifier PXD008119. We identified marginal differences between Mirasol PRT and Control PCs during storage. However, those significant changes at the proteome level were specifically related to the functional aspects previously described to affect platelets upon Mirasol PRT. In addition, the effect of Mirasol PRT on the platelet proteome appeared not to be exclusively due to an accelerated or enhanced PSL. In summary, semi-quantitative proteomics allows to discern between proteome changes due to Mirasol PRT or PSL, and proves to be a methodology suitable to phenotype platelets in an unbiased manner, in various physiological contexts.
Brailsford, S R; Tossell, J; Morrison, R; McDonald, C P; Pitt, T L
2018-05-24
Between February 2011 and December 2016, over 1·6 million platelet units, 36% pooled platelets, underwent bacterial screening prior to issue. Contamination rates for apheresis and pooled platelets were 0·02% and 0·07%, respectively. Staphylococcus aureus accounted for 21 contaminations, including four pooled platelets, one confirmed transfusion-transmitted infection (TTI) and three 'near-miss' incidents detected on visual inspection which were negative on screening. We describe follow-up investigations of 16 donors for skin carriage of S. aureus and molecular characterisation of donor and pack isolates. Units were screened by the BacT/ALERT 3D detection system. Contributing donors were interviewed and consent requested for skin and nasal swabbing. S. aureus isolates were referred for spa gene type and DNA macrorestriction profile to determine identity between carriage strains and packs. Donors of 10 apheresis and two pooled packs screen positive for S. aureus were confirmed as the source of contamination; seven had a history of skin conditions, predominantly eczema; 11 were nasal carriers. The 'near-miss' incidents were associated with apheresis donors, two donors harboured strains indistinguishable from the pack strain. The TTI was due to a screen-negative pooled unit, and a nasal isolate of one donor was indistinguishable from that in the unit. Staphylococcus aureus contamination is rare but potentially harmful in platelet units. Donor isolates showed almost universal correspondence in molecular type with pack isolates, thus confirming the source of contamination. The importance of visual inspection of packs prior to transfusion is underlined by the 'near-miss' incidents. © 2018 International Society of Blood Transfusion.
Platelet lysate formulations based on mucoadhesive polymers for the treatment of corneal lesions.
Sandri, Giuseppina; Bonferoni, Maria Cristina; Rossi, Silvia; Ferrari, Franca; Mori, Michela; Del Fante, Claudia; Perotti, Cesare; Scudeller, Luigia; Caramella, Carla
2011-02-01
Growth factors contained in platelet α-granules initiate and modulate tissue repair and are proposed for the treatment of soft and hard-tissue surgical conditions and in the management of non-healing wounds. Platelet lysate is a hemoderivative obtained from platelet-rich plasma and is capable of releasing a pool of growth factors. Many medical and surgical techniques have been proposed for the treatment of corneal lesions; management of these conditions remains problematic and healing with standard protocols is unattainable. The aim of this study was to develop formulations suitable for prolonging the contact of platelet lysate with the damaged cornea for the time necessary to exert a therapeutic effect. Two vehicles, one based on polyacrylic acid and one based on chitosan, were autoclaved and loaded with platelet lysate and the resultant formulations were characterized for rheology, mucoadhesion, vehicle compatibility and stability. The proliferation effect was tested on two cell culture types (rabbit corneal epithelial cells and fibroblasts). An in-vitro wound-healing test was performed on fibroblasts. In both cases the formulations were compared with platelet lysate diluted with saline at the same concentration. Both formulations maintained the rheological and mucoadhesive properties of the vehicles and the proliferative activity of platelet lysate. The chitosan formulation was able to significantly enhance epithelial cell growth even after storage of up to 2 weeks (in-use conditions), while the polyacrylic acid formulation was less efficient, probably due to the characteristics of the cell model used. The in-vitro wound-healing test performed on fibroblasts confirmed the differences between the two vehicles. The effect induced by the platelet lysate and chitosan formulation was faster than that of the polyacrylic acid formulation and complete in-vitro wound repair was achieved within 48 h. © 2010 The Authors. JPP © 2010 Royal Pharmaceutical Society.
Lotens, A; Najdovski, T; Cellier, N; Ernotte, B; Lambermont, M; Rapaille, A
2014-10-01
TACSI whole blood system is designed to combine primary and secondary processing of six whole blood bags into plasma units, buffy coat and red blood cell concentrates. The aim of this study was to investigate the specifications and in vitro storage parameters of blood components compared with standard centrifugation and separation processing. Whole blood bags, collected in CRC kits, were treated on a TACSI whole blood system. They were compared with whole blood bags collected in Composelect kits. In addition to routine quality control analyses, conservation studies were performed on red blood cell concentrates for 42 days and on plasma for 6 months. Platelets pools with five buffy coats were also created, and cellular contamination was evaluated. Red blood cell concentrates produced from TACSI whole blood met European quality requirements. For white blood cell count, one individual result exceeded 1 × 10(6) cells/unit. All plasma units fell within specifications for residual cellular contamination and storage parameters. The performances of the TACSI whole blood system allow for the preparation of low volume buffy coats with a recovery of 90% of whole blood platelets. Haemoglobin losses in TACSI BC are smaller, but this did not result in higher haemoglobin content of red cells. These BC are suitable for the production of platelet concentrates. From these in vitro data, red blood cell concentrates produced using TACSI whole blood are suitable for clinical use with a quality at least equivalent to the control group. © 2014 International Society of Blood Transfusion.
Johnson, Lacey; Tan, Shereen; Jenkins, Emily; Wood, Ben; Marks, Denese C
2018-04-01
Alternatives to room temperature storage of platelets (PLTs) are of interest to support blood banking logistics. The aim of this study was to compare the presence of biologic response modifiers (BRMs) in PLT concentrates stored under conventional room temperature conditions with refrigerated or cryopreserved PLTs. A three-arm pool-and-split study was carried out using buffy coat-derived PLTs stored in 30% plasma/70% SSP+. The three matched treatment arms were as follows: room temperature (20-24°C), cold (2-6°C), and cryopreserved (-80°C with DMSO). Liquid-stored PLTs were tested over a 21-day period, while cryopreserved PLTs were tested immediately after thawing and reconstitution in 30% plasma/70% SSP+ and after storage at room temperature. Coagulation factor activity was comparable between room temperature and cold PLTs, with the exception of protein S, while cryopreserved PLTs had reduced Factor (F)V and FVIII activity. Cold-stored PLTs retained α-granule proteins better than room temperature or cryopreserved PLTs. Cryopreservation resulted in 10-fold higher microparticle generation than cold-stored PLTs, but both groups contained significantly more microparticles than those stored at room temperature. The supernatant from both cold and cryopreserved PLTs initiated faster clot formation and thrombin generation than room temperature PLTs. Cold storage and cryopreservation alter the composition of the soluble fraction of stored PLTs. These differences in coagulation proteins, cytokines, and microparticles likely influence both the hemostatic capacity of the components and the auxiliary functions. © 2017 AABB.
Hermansky-Pudlak syndrome in a pregnant patient: a case report.
Harris-Glocker, Miranda; Thornburg, Loralei L; Pressman, Eva K
2013-01-01
Hermansky-Pudlak syndrome (HPS), a rare autosomal-recessive disorder encompassing multiple organs, is characterized by oculocutaneous albinism, platelet storage pool deficiency resulting in bleeding diathesis, and ceroid lipofuscin deposition which can lead to pulmonary fibrosis, colitis, cardiomyopathy and renal failure. Pregnancy in a patient with HPS can produce multiple complications such as peripartum hemorrhage and difficulties with administration of anesthesia, either regional or general. We present the case of a patient with HPS also complicated by spontaneous triplet pregnancy. A multidisciplinary approach, including the involvement of obstetric, anesthesia, and hematology teams, is the ideal for an HPS patient with the potential for multiple complications in the peripartum period.
Mageed, M; Ionita, C; Kissich, C; Brehm, W; Winter, K; Ionita, J-C
2015-01-01
To determine the influence of cryopreservation at two different temperatures on platelet concentration, growth factor (GF) levels and platelet activation parameters in equine ACP®; moreover, to determine if adding mechanical ACP® stimulation to freeze-thaw activation amplifies GF release from platelets. Firstly, blood from five horses was used to prepare ACP®. Platelet, platelet derived growth factor BB (PDGF-BB) and transforming growth factor β1 (TGF-β1) concentrations as well as mean platelet volume (MPV) and mean platelet component (MPC) were determined in fresh and corresponding ACP® samples after 2 months cryopreservation at -20 °C and -80 °C, respectively. Secondly, ACP® was prepared from blood of nine horses. Half of ACP® was activated using one freeze-thaw-cycle at -20 °C, whereas the rest was first vortexed. Their PDGF-BB and TGF-β1 concentrations were subsequently determined. Platelet concentration significantly decreased after -80 °C cryopreservation. PDGF-BB level augmented significantly after both storage methods, whereas TGF-β1 concentration was not significantly altered. MPV significantly increased after -20 °C cryopreservation. Both storage regimens induced a significant MPC decrease. No significant differences in GF concentrations between the vortexed and non-vortexed samples were detected. Both cryopreservation methods induced platelet activation, but storage at -80 °C apparently harmed the platelets without generating higher GF release than -20 °C. The mechanical stimulation process could not enhance GF release in subsequently frozen-thawed ACP®. Storage of ACP® at -20 °C could be useful in equine practice, but, before this procedure can be recommended, further qualitative tests are needed. The mechanical stimulation technique should be adjusted in order to increase platelet activation.
Gitz, Eelo; Koekman, Cornelis A; van den Heuvel, Dave J.; Deckmyn, Hans; Akkerman, Jan W.; Gerritsen, Hans C.; Urbanus, Rolf T.
2012-01-01
Background Storing platelets for transfusion at room temperature increases the risk of microbial infection and decreases platelet functionality, leading to out-date discard rates of up to 20%. Cold storage may be a better alternative, but this treatment leads to rapid platelet clearance after transfusion, initiated by changes in glycoprotein Ibα, the receptor for von Willebrand factor. Design and Methods: We examined the change in glycoprotein Ibα distribution using Förster resonance energy transfer by time-gated fluorescence lifetime imaging microscopy. Results Cold storage induced deglycosylation of glycoprotein Ibα ectodomain, exposing N-acetyl-Dglucosamine residues, which sequestered with GM1 gangliosides in lipid rafts. Raft-associated glycoprotein Ibα formed clusters upon binding of 14-3-3ζ adaptor proteins to its cytoplasmic tail, a process accompanied by mitochondrial injury and phosphatidyl serine exposure. Cold storage left glycoprotein Ibα surface expression unchanged and although glycoprotein V decreased, the fall did not affect glycoprotein Ibα clustering. Prevention of glycoprotein Ibα clustering by blockade of deglycosylation and 14-3-3ζ translocation increased the survival of cold-stored platelets to above the levels of platelets stored at room temperature without compromising hemostatic functions. Conclusions We conclude that glycoprotein Ibα translocates to lipid rafts upon cold-induced deglycosylation and forms clusters by associating with 14-3-3ζ. Interference with these steps provides a means to enable cold storage of platelet concentrates in the near future. PMID:22733027
Vamvakas, Eleftherios C
2011-01-01
The risks of known and emerging transfusion-transmitted infections (TTIs) from reducing the current lifetime blood donation deferral for men who have had sex with men (MSM) to 1 or 5 years were compared to the risk from continuing to transfuse in the United States 12.5% of platelet doses as pooled whole-blood-derived (rather than single-donor) platelets. Assumptions made in mathematical models and blood donor/transfusion studies of the risks of TTIs since 2000 were evaluated. The number of HIV, hepatitis B virus, or hepatitis C virus TTIs from reducing the MSM deferral to 1 year is, respectively, 0.88, 2.94, or 66.9, many more than 10 times smaller than the risk from pooled platelets. If erroneous release of HIV-positive units (a risk independent of a donor's source of infection) is not considered, the MSM risk is 1 HIV-infectious donation per 17 to 56 million MSM donations. Any purportedly increased risk of human herpesvirus-8 transmission from MSM donors is far smaller than the risk of transfusion-associated sepsis from pooled platelets. Single-donor platelets from MSM after 5 years' abstinence are as safe or 5 times safer than our current pooled platelets--if the next TTI to emerge were transmitted, respectively, sexually or by another route. Thus, acceptance of MSM as blood donors after 1 or 5 years' abstinence may result in a postulated increase in risk that is so much smaller than the currently tolerated transfusion risk and so small in absolute terms that the ethical question of fairness to the MSM group justifies the change in policy. Copyright © 2011 Elsevier Inc. All rights reserved.
Browning, Lucy M; Walker, Celia G; Mander, Adrian P; West, Annette L; Madden, Jackie; Gambell, Joanna M; Young, Stephen; Wang, Laura; Jebb, Susan A; Calder, Philip C
2012-10-01
Estimation of the intake of oily fish at a population level is difficult. The measurement of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in biological samples may provide a useful biomarker of intake. We identified the most appropriate biomarkers for the assessment of habitual oily fish intake and changes in intake by elucidating the dose- and time-dependent response of EPA and DHA incorporation into various biological samples that represent roles in fatty acid transport, function, and storage. This was a double-blind, randomized, controlled intervention trial in 204 men and women that lasted 12 mo. EPA and DHA capsules were provided in a manner to reflect sporadic consumption of oily fish (ie, 1, 2, or 4 times/wk). EPA and DHA were assessed at 9 time points over 12 mo in 9 sample types (red blood cells, mononuclear cells, platelets, buccal cells, adipose tissue, plasma phosphatidylcholine, triglycerides, cholesteryl esters, and nonesterified fatty acids). A dose response (P < 0.05) was observed for EPA and DHA in all pools except for red blood cell EPA (P = 0.057). EPA and DHA measures in plasma phosphatidylcholine and platelets were best for the discrimination between different intakes (P < 0.0001). The rate of incorporation varied between sample types, with the time to maximal incorporation ranging from days (plasma phosphatidylcholine) to months (mononuclear cells) to >12 mo (adipose tissue). Plasma phosphatidylcholine EPA plus DHA was identified as the most suitable biomarker of acute changes in EPA and DHA intake, and platelet and mononuclear cell EPA plus DHA were the most suitable biomarkers of habitual intake.
1992-11-02
linking agents. Water replacement molecules, such as trehalose or sucrose at concentrations up to 1 M, did not suffice to stabilize platelets for...concentration of albumin (5% w/v) or trehalose (250 mM - I M) is required in the final suspension prior to freeze-drying to ensure platelet yield and...structurally or functionally intact platelets post- rehydration; the presence of albumin or trehalose raises this yield to 90-100%. Lvophilization and Storage
The behaviour of platelets in natural diamonds and the development of a new mantle thermometer
NASA Astrophysics Data System (ADS)
Speich, L.; Kohn, S. C.; Bulanova, G. P.; Smith, C. B.
2018-05-01
Platelets are one of the most common defects occurring in natural diamonds but their behaviour has not previously been well understood. Recent technical advances, and a much improved understanding of the correct interpretation of the main infrared (IR) feature associated with platelets (Speich et al. 2017), facilitated a systematic study of platelets in 40 natural diamonds. Three different types of platelet behaviour were identified here. Regular diamonds show linear correlations between both B-centre concentrations and platelet density and also between platelet size and platelet density. Irregular diamonds display reduced platelet density due to platelet breakdown, anomalously large or small platelets and a larger platelet size distribution. These features are indicative of high mantle storage temperatures. Finally, a previously unreported category of subregular diamonds is defined. These diamonds experienced low mantle residence temperatures and show smaller than expected platelets. Combining the systematic variation in platelet density with temperatures of mantle storage, determined by nitrogen aggregation, we can demonstrate that platelet degradation proceeds at a predictable rate. Thus, in platelet-bearing diamonds where N aggregation is complete, an estimate of annealing temperature can now be made for the first time.
Saito, Shunnichi; Nollet, Kenneth E; Ngoma, Alain M; Ono, Takako; Ohto, Hitoshi
2018-02-01
Storage lesion, including microparticle formation, has been partially characterised in whole blood, but not in all combinations of pre-storage leucofiltration and/or irradiation. Single-donor whole blood products were processed into four subunits: with and without leucofiltration, with and without X-irradiation (25 Gy). Platelet-, leucocyte-, and erythrocyte-derived microparticles and free haemoglobin were measured periodically throughout 42 days of storage. Pre-storage leucofiltration substantially reduced platelet- and leucocyte-derived microparticle counts throughout storage. Irradiation, in contrast, had no significant effect on microparticle counts. A gate for all microparticles showed a substantial time-dependent increase in unfiltered whole blood. A time-dependent increase in free haemoglobin was greatest in unfiltered, irradiated whole blood. This study indicates that leucofiltration can prevent the formation of leucocyte- and platelet-derived microparticles, and might reduce haemolysis in irradiated whole blood, either by removing factors that provoke haemolysis, or by selective retention of senescent or effete red cells most prone to haemolysis.
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.
Prolonging shelf-life of platelets by low-level laser
NASA Astrophysics Data System (ADS)
Zhang, Qi; Lu, Min; Wu, Mei X.
2018-02-01
It remains significant challenges to extend a shelf life of platelets beyond the conventional five days. Unlike red blood cells that can be stored at 4°C for a few weeks, platelets are stored at room temperature only, which results in a gradual loss of their quality owing to a switch of energy metabolism from aerobic oxidative phosphorylation toward anaerobic glycolysis. Given the well-documented beneficial effect of near infrared low-level laser (LLL) on mitochondrial functions in a variety of cells under stress, we explored a potential for LLL to extend the shelf life of platelets beyond the five days. We found that exposure of a platelet-containing storage bag to LLL at 830nm at 0.5J/cm2 prior to storage could significantly retain a pH value and viability of the platelets stored within the bag under a standard condition for eight days with improved quality compared to those platelets stored similarly for five days in controls. LLL inhibited reactive oxygen species (ROS) and lactate production, but sustained ATP production, mitochondrial membrane potential, and morphology in the stored platelets. While preserving their metabolic activity, LLL didn't activate platelets but increased their aggregation capacity and in vivo survival as suggested by similar levels of surface CD62p expression and enhanced agonist-induced aggregation and recovery following infusion in the presence compared to the absence of LLL treatment. This simple, addition-free, cost-effective, noninvasive laser illumination can be readily incorporated into the current platelet storage system to prolong shelf life of platelets with improved quality of stored platelets.
Maloney, James P; Ambruso, Daniel R; Voelkel, Norbert F; Silliman, Christopher C
The occurrence of non-hemolytic transfusion reactions is highest with platelet and plasma administration. Some of these reactions are characterized by endothelial leak, especially transfusion related acute lung injury (TRALI). Elevated concentrations of inflammatory mediators secreted by contaminating leukocytes during blood product storage may contribute to such reactions, but platelet-secreted mediators may also contribute. We hypothesized that platelet storage leads to accumulation of the endothelial permeability mediator vascular endothelial growth factor (VEGF), and that intravascular administration of exogenous VEGF leads to extensive binding to its lung receptors. Single donor, leukocyte-reduced apheresis platelet units were sampled over 5 days of storage. VEGF protein content of the centrifuged supernatant was determined by ELISA, and the potential contribution of VEGF from contaminating leukocytes was quantified. Isolated-perfused rat lungs were used to study the uptake of radiolabeled VEGF administered intravascularly, and the effect of unlabeled VEGF on lung leak. There was a time-dependent release of VEGF into the plasma fraction of the platelet concentrates (62 ± 9 pg/ml on day one, 149 ± 23 pg/ml on day 5; mean ± SEM, p<0.01, n=8) and a contribution by contaminating leukocytes was excluded. Exogenous 125I-VEGF bound avidly and specifically to the lung vasculature, and unlabeled VEGF in the lung perfusate caused vascular leak. Rising concentrations of VEGF occur during storage of single donor platelet concentrates due to platelet secretion or disintegration, but not due to leukocyte contamination. Exogenous VEGF at these concentrations rapidly binds to its receptors in the lung vessels. At higher VEGF concentrations, VEGF causes vascular leak in uninjured lungs. These data provide further evidence that VEGF may contribute to the increased lung permeability seen in TRALI associated with platelet products.
Stochastic response of human blood platelets to stimulation of shape changes and secretion.
Deranleau, D A; Lüthy, R; Lüscher, E F
1986-01-01
Stopped-flow turbidimetric data indicate that platelets stimulated with low levels of thrombin undergo a shape transformation from disc to "sphere" to smaller spiny sphere that is indistinguishable from the shape change induced by ADP through different membrane receptor sites and a dissimilar receptor trigger mechanism. Under conditions where neither secretion nor aggregation occur, the extinction coefficients for total scattering by each of the three platelet forms are independent of the stimulus applied, and both reaction mechanisms can be described as stochastic (Poisson) processes in which the rate constant for the formation of the transient species is equal to the rate constant for its disappearance. This observation is independent of the shape assignment, and as the concentration of thrombin is increased and various storage organelles secrete increasing amounts of their contents into the external medium, the stochastic pattern persists. Progressively larger decreases in the extinction coefficients of the intermediate and final platelet forms, over and above those that reflect shape alterations alone, accompany or parallel the reaction induced by the higher thrombin concentrations. The excess turbidity decrease observed when full secretion occurs can be wholly accounted for by a decrease in platelet volume equal in magnitude to the fraction of the total platelet volume occupied by alpha granules. Platelet activation, as reported by the whole body light scattering of either shape changes alone or shape changes plus parallel (but not necessarily also stochastic) alpha granule secretion, thus manifests itself as a random series of transient events conceivably with its origins in the superposition of a set of more elementary stochastic processes that could include microtubule depolymerization, actin polymerization, and possibly diffusion. Although the real nature of the control mechanism remains obscure, certain properties of pooled stochastic processes suggest that a reciprocal connection between microtubule fragmentation and the assembly of actin-containing pseudopodal structures and contractile elements--processes that may exhibit reciprocal requirements for calcium--might provide a hypothetical basis for a rate-limiting step. PMID:3457375
Initial blood storage experiment
NASA Technical Reports Server (NTRS)
Surgenor, Douglas MACN.
1988-01-01
The possibility of conducting experiments with the formed elements of the blood under conditions of microgravity opens up important opportunities to improve the understanding of basic formed element physiology, as well as, contribution to improved preservation of the formed elements for use in transfusion. The physiological, biochemical, and physical changes of the membrane of the erythrocyte, platelet, and leukocyte was studied during storage under two specific conditions: standard blood bank conditions and microgravity, utilizing three FDA approved plastic bags. Storage lesions; red cell storage on Earth; platelet storage on Earth; and leukocyte storage Earth were examined. The interaction of biomaterials and blood cells was studied during storage.
Johnson, Lacey; Schubert, Peter; Tan, Shereen; Devine, Dana V; Marks, Denese C
2016-02-01
The storage of platelets (PLTs) in additive solution (AS) may facilitate improved PLT quality and possibly extension of the PLT shelf life. A minimum amount of plasma is required when PLTs are stored in AS, as a source of glucose. The aim of this study was to assess the effect of reducing the plasma carryover to 20% on PLT quality when stored in SSP+ for an extended period. Using a pool-and-split design, buffy coat-derived PLTs were stored in either 30% plasma/SSP+ or 20% plasma/SSP+. In vitro analyses were carried out to Day 10. Metabolites and markers of PLT activation and apoptosis were measured using a blood gas analyzer and flow cytometry. PLT apoptotic protein expression was investigated by Western blotting. Glucose exhaustion occurred in the 20% plasma group between Day 7 and Day 10. The surface expression of P-selectin and PAC-1 was comparable on Day 10 in both groups, suggesting that the PLTs were not activated. However, the exposure of phosphatidylserine and the number of phosphatidylserine-positive microparticles were significantly higher in the 20% group on Day 10. The expression of the proapoptotic proteins Bak, Bax, and cleaved caspase-3 were higher in the 20% plasma group by Day 7 of storage, compared to the 30% plasma group. Exhaustion of glucose was associated with a proapoptotic phenotype. Results such as these should be considered before extending the PLT shelf life beyond 7 days, particularly when stored in ASs lacking glucose with low plasma carryover. © 2015 AABB.
Saito, Shunnichi; Ngoma, Alain M.; Ono, Takako; Ohto, Hitoshi
2018-01-01
Background Storage lesion, including microparticle formation, has been partially characterised in whole blood, but not in all combinations of pre-storage leucofiltration and/or irradiation. Materials and methods Single-donor whole blood products were processed into four subunits: with and without leucofiltration, with and without X-irradiation (25 Gy). Platelet-, leucocyte-, and erythrocyte-derived microparticles and free haemoglobin were measured periodically throughout 42 days of storage. Results Pre-storage leucofiltration substantially reduced platelet- and leucocyte-derived microparticle counts throughout storage. Irradiation, in contrast, had no significant effect on microparticle counts. A gate for all microparticles showed a substantial time-dependent increase in unfiltered whole blood. A time-dependent increase in free haemoglobin was greatest in unfiltered, irradiated whole blood. Discussion This study indicates that leucofiltration can prevent the formation of leucocyte- and platelet-derived microparticles, and might reduce haemolysis in irradiated whole blood, either by removing factors that provoke haemolysis, or by selective retention of senescent or effete red cells most prone to haemolysis. PMID:27893349
Sergeeva, N S; Shanskii, Ya D; Sviridova, I K; Karalkin, P A; Kirsanova, V A; Akhmedova, S A; Kaprin, A D
2016-11-01
Platelet lysate prepared from donor platelet concentrate and pooled according to a developed technique stimulates migration of multipotent mesenchymal stromal cells of the human adipose tissue and promotes healing of the monolayer defect in cultures of human fibroblasts and multipotent mesenchymal stromal cells in vitro in concentrations close those of fetal calf serum (5-10%). Lysate of platelets from platelet-rich rat blood plasma stimulated healing of the skin defect by promoting epithelialization and granulation tissue formation. The regenerative properties of platelet lysate in vivo increased with increasing its concentration.
Störmer, M; Cassens, U; Kleesiek, K; Dreier, J
2007-02-01
Bacteria show differences in their growth kinetics depending on the type of blood component. On to storage at 22 degrees C, platelet concentrates (PCs) seem to be more prone to bacterial multiplication than red cell concentrates. Knowledge of the potential for bacterial proliferation in blood components, which are stored at a range of temperatures, is essential before considering implementation of a detection strategy. The efficacy of bacterial detection was determined, using real-time reverse transcriptase-polymerase chain reaction (RT-PCR), following bacterial growth in blood components obtained from a deliberately contaminated whole-blood (WB) unit. Cultivation was used as the reference method. WB was spiked with 2 colony-forming units mL(-1)Staphylococcus epidermidis or Klebsiella pneumoniae, kept for 15 h at room temperature and component preparation was processed. Samples were drawn, at intervals throughout the whole separation process, from each blood component. Nucleic acids were extracted using an automated high-volume extraction method. The 15-h storage revealed an insignificant increase in bacterial titre. No bacterial growth was detected in red blood cell or plasma units. K. pneumoniae showed rapid growth in the pooled PC and could be detected immediately after preparation using RT-PCR. S. epidermidis grew slowly and was detected 24 h after separation. These experiments show that sampling is indicative at 24 h after preparation of PCs at the earliest to minimize the sampling error.
Microparticles variability in fresh frozen plasma: preparation protocol and storage time effects
Kriebardis, Anastasios G.; Antonelou, Marianna H.; Georgatzakou, Hara T.; Tzounakas, Vassilis L.; Stamoulis, Konstantinos E.; Papassideri, Issidora S.
2016-01-01
Background Extracellular vesicles or microparticles exhibiting procoagulant and thrombogenic activity may contribute to the haemostatic potential of fresh frozen plasma. Materials and methods Fresh frozen plasma was prepared from platelet-rich plasma at 20 °C (Group-1 donors) or directly from whole blood at 4 °C (Group-2 donors). Each unit was aseptically divided into three parts, stored frozen for specific periods of time, and analysed by flow cytometry for procoagulant activity immediately after thaw or following post-thaw storage for 24 h at 4 °C. Donors’ haematologic, biochemical and life-style profiles as well as circulating microparticles were analysed in parallel. Results Circulating microparticles exhibited a considerable interdonor but not intergroup variation. Fresh frozen plasma units were enriched in microparticles compared to plasma in vivo. Duration of storage significantly affected platelet- and red cell-derived microparticles. Fresh frozen plasma prepared directly from whole blood contained more residual platelets and more platelet-derived microparticles compared to fresh frozen plasma prepared from platelet-rich plasma. Consequently, there was a statistically significant difference in total, platelet- and red cell-derived microparticles between the two preparation protocols over storage time in the freezer. Preservation of the thawed units for 24 h at 4 °C did not significantly alter microparticle accumulation. Microparticle accumulation and anti-oxidant capacity of fresh frozen plasma was positively or negatively correlated, respectively, with the level of circulating microparticles in individual donors. Discussion The preparation protocol and the duration of storage in the freezer, independently and in combination, influenced the accumulation of microparticles in fresh frozen plasma units. In contrast, storage of thawed units for 24 h at 4 °C had no significant effect on the concentration of microparticles. PMID:27136430
Microparticles variability in fresh frozen plasma: preparation protocol and storage time effects.
Kriebardis, Anastasios G; Antonelou, Marianna H; Georgatzakou, Hara T; Tzounakas, Vassilis L; Stamoulis, Konstantinos E; Papassideri, Issidora S
2016-05-01
Extracellular vesicles or microparticles exhibiting procoagulant and thrombogenic activity may contribute to the haemostatic potential of fresh frozen plasma. Fresh frozen plasma was prepared from platelet-rich plasma at 20 °C (Group-1 donors) or directly from whole blood at 4 °C (Group-2 donors). Each unit was aseptically divided into three parts, stored frozen for specific periods of time, and analysed by flow cytometry for procoagulant activity immediately after thaw or following post-thaw storage for 24 h at 4 °C. Donors' haematologic, biochemical and life-style profiles as well as circulating microparticles were analysed in parallel. Circulating microparticles exhibited a considerable interdonor but not intergroup variation. Fresh frozen plasma units were enriched in microparticles compared to plasma in vivo. Duration of storage significantly affected platelet- and red cell-derived microparticles. Fresh frozen plasma prepared directly from whole blood contained more residual platelets and more platelet-derived microparticles compared to fresh frozen plasma prepared from platelet-rich plasma. Consequently, there was a statistically significant difference in total, platelet- and red cell-derived microparticles between the two preparation protocols over storage time in the freezer. Preservation of the thawed units for 24 h at 4 °C did not significantly alter microparticle accumulation. Microparticle accumulation and anti-oxidant capacity of fresh frozen plasma was positively or negatively correlated, respectively, with the level of circulating microparticles in individual donors. The preparation protocol and the duration of storage in the freezer, independently and in combination, influenced the accumulation of microparticles in fresh frozen plasma units. In contrast, storage of thawed units for 24 h at 4 °C had no significant effect on the concentration of microparticles.
Larsen, C P; Ezligini, F; Hermansen, N O; Kjeldsen-Kragh, J
2005-02-01
Approximately 1 in every 2000 units of platelets is contaminated with bacteria. The BacT/ALERT automated blood culture system can be used to screen platelet concentrates (PCs) for bacterial contamination. Data were collected from May 1998 until May 2004. The number of PCs tested during this period was 36 896, most of which were produced from pools of four buffy-coats. On the day following blood collection or platelet apheresis, a 5-10 ml sample of the PC was aseptically transferred to a BacT/ALERT culture bottle for detection of aerobic bacteria. The sample was monitored for bacterial growth during the entire storage period of the PC (6.5 days). When a positive signal was generated, the culture bottle, the PC and the erythrocyte concentrates were tested for bacterial growth. In order to determine the frequency of false-negative BacT/ALERT signals, 1061 outdated PCs were tested during the period from May 2002 to May 2004. Eighty-eight positive signals were detected by the BacT/ALERT system, of which 12 were interpreted as truly positive. Fourteen signals were interpreted as truly false positive. Thirty-three signals were interpreted to be probably false positive. Two of 1061 outdated units tested positive, and Bacillus spp. and Staphylococcus epidermidis, respectively, were isolated from these PCs. Between 0.03% and 0.12% of the PCs were contaminated with bacteria. BacT/ALERT is an efficient tool for monitoring PCs for bacterial contamination; however, it is important to realize that false-negative results may occur.
Platelets in blood stored in untreated and siliconed glass bottles and plastic bags
Kissmeyer-Nielsen, F.; Madsen, C. B.; Nedergaard, Jytte
1961-01-01
Platelet survival was determined using untreated and siliconed glass bottles and plastic bags (Fenwal) for collecting and storing blood. The platelets were tagged in vivo with P32 in six polycythaemic patients undergoing treatment with P32. The results showed that fresh ACD blood collected in untreated glass, siliconed glass, and plastic gave the same recovery of platelets in the recipients. The use of EDTA (Fenwal formula) as anticoagulant gave results inferior to those obtained with blood using ACD as anticoagulant. Even after storage up to 24 hours in untreated glass bottles (ordinary bank blood) a satisfactory recovery of platelets was observed. After storage for 72 hours the recovery was less but not negligible. PMID:14456481
The Non-Hemostatic Aspects of Transfused Platelets
Sut, Caroline; Tariket, Sofiane; Aubron, Cécile; Aloui, Chaker; Hamzeh-Cognasse, Hind; Berthelot, Philippe; Laradi, Sandrine; Greinacher, Andreas; Garraud, Olivier; Cognasse, Fabrice
2018-01-01
Platelets transfusion is a safe process, but during or after the process, the recipient may experience an adverse reaction and occasionally a serious adverse reaction (SAR). In this review, we focus on the inflammatory potential of platelet components (PCs) and their involvement in SARs. Recent evidence has highlighted a central role for platelets in the host inflammatory and immune responses. Blood platelets are involved in inflammation and various other aspects of innate immunity through the release of a plethora of immunomodulatory cytokines, chemokines, and associated molecules, collectively termed biological response modifiers that behave like ligands for endothelial and leukocyte receptors and for platelets themselves. The involvement of PCs in SARs—particularly on a critically ill patient’s context—could be related, at least in part, to the inflammatory functions of platelets, acquired during storage lesions. Moreover, we focus on causal link between platelet activation and immune-mediated disorders (transfusion-associated immunomodulation, platelets, polyanions, and bacterial defense and alloimmunization). This is linked to the platelets’ propensity to be activated even in the absence of deliberate stimuli and to the occurrence of time-dependent storage lesions. PMID:29536007
Wagner, Stephen J; Myrup, Andrew; Awatefe, Helen; Thompson-Montgomery, Dedeene; Hirayama, Junichi; Skripchenko, Andrey
2008-12-01
Extensive periods without agitation can occasionally occur during platelet (PLT) shipment and can affect PLT quality during 5- to 7-day storage. The use of buffer-containing PLT additive solutions (ASs) may better preserve PLT quality during storage by maintaining PLT pH and other in vitro variables. A newly described bicarbonate-containing AS, M-sol, was compared to plasma for preservation of whole blood-derived PLT concentrates in which a 30-hour interruption of agitation was included. ABO-identical PLT-rich plasma intermediate products were pooled in sets of four, split, and centrifuged with subsequent plasma expression (n = 12). Two units were resuspended with M-sol AS to produce a 70 percent solution/30 percent plasma PLT concentrate; 2 units were resuspended in 100 percent plasma. One M-sol resuspended unit and 1 plasma unit were held on a laboratory bench in a standard shipping box for 30 hours between Day 2 and Day 3, while the other M-sol and plasma unit were continuously agitated. Standard in vitro testing for PLT quality variables on each set of 4 units was performed during storage (n = 12). Interrupting agitation of PLTs suspended in M-sol resulted in less of a pH decrement during storage than that of PLTs suspended in 100 percent plasma. On Days 5 and 7, the pH differences between M-sol and plasma units were 0.56 and 0.75 pH units, respectively (p < 0.0003). In addition, PLTs suspended in M-sol and subjected to an interruption of agitation had lesser Day 7 CD62+ cells, glucose utilization, and lactate production and greater hypotonic stress response, morphology, swirling, and aggregation response than those suspended in plasma (p = 0.005). The in vitro properties of PLTs suspended in 70 percent M-sol/30 percent plasma and subjected to a 30-hour interruption of agitation are better maintained during 7-day storage than those of matched units suspended in plasma.
Hlavac, N; Lasta, C S; Dalmolin, M L; Lacerda, L A; de Korte, D; Marcondes, N A; Terra, S R; Fernandes, F B; González, F H D
2017-11-15
Platelet transfusion therapy poses many challenges in veterinary clinical practice. Lack of readily available blood donors, short shelf-life, and inability to administer a sufficient number of platelets to meet a dog's transfusion need are the major difficulties encountered. Platelet additive solutions are already in use at American and European human blood banks, showing to be a realistic alternative. This study compares the in vitro platelet function in plasma, Composol, or SSP+ during storage for 13 days. Platelet rich plasma-platelet concentrate with 35% plasma and 65% platelet additive solutions (Composol or SSP+) and a control group (100% plasma) were prepared. Swirling, platelet count, blood gases, metabolic variables, platelet activation markers, and apoptosis markers were analyzed on days 1, 5, 9 and 13. Swirling was well preserved and pH was acceptable (> 6.2) during storage for all platelet additive solutions units until day 9. SSP + units showed more stable pH and metabolic variables until day 13. Platelets in plasma showed higher glucose consumption than in Composol or in SSP+. The platelet additive solutions units showed better platelet metabolism maintenance, reduced glucose consumption and lactate production. The apoptotic markers were still low for 9 days in platelet concentrates with platelet additive solutions, suggesting the possibility to extend the shelf life with the use of SSP+ or Composol. Our findings suggest that the uses of Composol and SSP+ in canine platelet concentrates are potential alternatives in veterinary blood banks.
Electrospun silk fibroin fibers for storage and controlled release of human platelet lysate.
Pignatelli, Cataldo; Perotto, Giovanni; Nardini, Marta; Cancedda, Ranieri; Mastrogiacomo, Maddalena; Athanassiou, Athanassia
2018-04-17
Human platelet lysate (hPL) is a pool of growth factors and cytokines able to induce regeneration of different tissues. Despite its good potentiality as therapeutic tool for regenerative medicine applications, hPL has been only moderately exploited in this field. A more widespread adoption has been limited because of its rapid degradation at room temperature that decreases its functionality. Another limiting factor for its extensive use is the difficulty of handling the hPL gels. In this work, silk fibroin-based patches were developed to address several points: improving the handling of hPL, enabling their delivery in a controlled manner and facilitating their storage by creating a device ready to use with expanded shelf life. Patches of fibroin loaded with hPL were synthesized by electrospinning to take advantage of the fibrous morphology. The release kinetics of the material was characterized and tuned through the control of fibroin crystallinity. Cell viability assays, performed with primary human dermal fibroblasts, demonstrated that fibroin is able to preserve the hPL biological activity and prolong its shelf-life. The strategy of storing and preserving small active molecules within a naturally-derived, protein-based fibrous scaffold was successfully implemented, leading to the design of a biocompatible device, which can potentially simplify the storage and the application of the hPL on a human patient, undergoing medical procedures such as surgery and wound care. Human platelets lysate (hPL) is a mixture of growth factors and cytokines able to induce the regeneration of damaged tissues. This study aims at enclosing hPL in a silk fibroin electrospun matrix to expand its utilization. Silk fibroin showed the ability to preserve the hPL activity at temperature up to 60 °C and the manipulation of fibroin's crystallinity provided a tool to modulate the hPL release kinetic. This entails the possibility to fabricate the hPL silk fibroin patches in advance and store them, resulting in an easy and fast accessibility and an expanded use of hPL for wound healing. Copyright © 2018 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
Schubert, Peter; Thon, Jonathan N.; Walsh, Geraldine M.; Chen, Cindy H.I.; Moore, Edwin D.; Devine, Dana V.; Kast, Juergen
2015-01-01
BACKGROUND The term platelet storage lesion (PSL) describes the structural and biochemical changes in platelets (PLTs) during storage. These are typified by alterations of morphologic features and PLT metabolism leading to reduced functionality and hence reduced viability for transfusion. While the manifestations of the storage lesion are well characterized, the biochemical pathways involved in the initiation of this process are unknown. STUDY DESIGN AND METHODS A complementary proteomic approach has recently been applied to analyze changes in the PLT proteome during storage. By employing stringent proteomic criteria, 12 proteins were identified as significantly and consistently changing in relative concentration over a 7-day storage period. Microscopy, Western blot analysis, flow cytometry, and PLT functionality analyses were used to unravel the involvement of a subset of these 12 proteins, which are connected through integrin signaling in one potential signaling pathway underlying storage lesion development. RESULTS Microscopic analysis revealed changes in localization of glycoprotein IIIa, Rap1, and talin during storage. Rap1 activation was observed to correlate with expression of the PLT activation marker CD62P. PLTs incubated for 7 days with the PI3-kinase inhibitor LY294002 showed diminished Rap1 activation as well as a moderate reduction in integrin αIIbβ3 activation and release of α-granules. Furthermore, this inhibitor seemed to improve PLT integrity and quality during storage as several in vitro probes showed a deceleration of PLT activation. CONCLUSION These results provide the first evidence for a signaling pathway mediating PSL in which PI3-kinase–dependent Rap1 activation leads to integrin αIIbβ3 activation and PLT degranulation. PMID:19497060
Granat, Fanny; Geffré, Anne; Bourgès-Abella, Nathalie; Braun, Jean-Pierre; Trumel, Catherine
2013-06-01
In veterinary medicine a complete blood cell count (CBC) cannot always be performed within 24 h as usually recommended, particularly for specimens shipped to a reference laboratory. This raises the question of the stability of the variables, especially in ethylenediamine tetra-acetic acid (EDTA) feline blood specimens, known to be prone to in vitro platelet aggregation. Citrate, theophylline, adenosine and dipyridamole (CTAD) has been reported to limit platelet aggregation in feline blood specimens. The aim of this study was to measure the stability of the haematological variables and the platelet aggregation score in EDTA and EDTA plus CTAD (EDCT) feline blood specimens during 48 h of storage at room temperature. Forty-six feline EDTA and EDCT blood specimens were analysed with a Sysmex XT-2000iV analyser, and the platelet count and score of platelet aggregation were estimated immediately and after 24 and 48 h of storage. A significant increase in mean corpuscular volume, haematocrit, reticulocyte and eosinophil counts, and a significant decrease in mean corpuscular haemoglobin concentration and monocyte count were observed. Haemoglobin, mean corpuscular haemoglobin, and red blood cell, white blood cell, neutrophil and lymphocyte counts remained stable. Changes in reticulocyte indexes with time (low fluorescence ratio, medium fluorescence ratio, high fluorescence ratio and immature reticulocyte fraction) were not significant. Changes were generally more pronounced in EDTA than in EDCT. Platelet aggregation decreased markedly in initially highly aggregated EDTA specimens, and increased slightly in initially non- or mildly-aggregated EDTA or EDCT specimens. Platelet counts increased and decreased, or remained stable, respectively. CTAD can reduce storage-induced changes of the haematological variables in feline samples, thus improving the reliability of a CBC and limiting clinical misinterpretations.
1994-10-27
paraformaidehyde in 500 mM Trehalose stored desiccated at RT, 4* C, or at -70,° C. Neither prep maintained good morphology at any temperature, and there...platelets, or para-platelets dried in Trehalose are as susceptible to loss of integrity over time as other preps. Our platelet handling techniques have
George, Mitchell J; Bynum, James; Nair, Prajeeda; Cap, Andrew P; Wade, Charles E; Cox, Charles S; Gill, Brijesh S
2018-07-01
The purpose of this review is to explore the relationship between platelet bioenergetics and biomechanics and how this relationship affects the clinical interpretation of platelet function devices. Recent experimental and technological advances highlight platelet bioenergetics and biomechanics as alternative avenues for collecting clinically relevant data. Platelet bioenergetics drive energy production for key biomechanical processes like adhesion, spreading, aggregation, and contraction. Platelet function devices like thromboelastography, thromboelastometry, and aggregometry measure these biomechanical processes. Platelet storage, stroke, sepsis, trauma, or the activity of antiplatelet drugs alters measures of platelet function. However, the specific mechanisms governing these alterations in platelet function and how they relate to platelet bioenergetics are still under investigation.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Gear, A.R.L.; Hallam, T.J.
1986-03-01
Interest in phosphatidylinositol metabolism has been greatly stimulated by the findings that diglyceride and inositol phosphates may serve as second messengers in modulating cellular function. Formation of 1,4,5-inositol trisphosphate (IP/sub 3/), in particular, has been linked to mobilization of intracellular calcium in a number of cell types. The authors have examined the ability of IP/sub 3/ to mobilize calcium in human platelets permeabilized by either saponin or high-voltage discharge. Saponin at 15 ..mu..g/ml effectively permeabilized platelets to exogenous inositol 1,4,5-trisphosphate which released bound (/sup 45/Ca) within 1 min and with a Ka of 7.4 +/- 4.1 ..mu..M. A small (25%)more » azide-sensitive pool was also responsive to inositol trisphosphate. The calcium pools were completely discharged by A-23187 and the ATP-dependent uptake was prevented by dinitrophenol. In contrast to the result with saponin, platelets accessed by high-voltage discharge were insensitive to challenge by inositol 1,4,5-trisphosphate. The data suggest that while inositol 1,4,5-trisphosphate can rapidly mobilize platelet calcium, the ability to demonstrate this depends on the method of permeabilization.« less
[Allergic transfusion reactions in a patient with multiple food allergies].
Strobel, E; Schöniger, M; Münz, M; Hiefinger-Schindlbeck, R
2012-07-01
A 13-year-old girl with an osteosarcoma was treated by surgery and chemotherapy. During three transfusions of apheresis platelet concentrates allergic reactions occurred, partly in spite of premedication with an antihistamine and a corticoid. As the patient declared to be allergic to some foods, in-vitro tests for allergen-specific IgE antibodies were performed and showed markedly positive results for specific IgE to carrot and celery, less so to hazelnut, peanut and a lot of other food antigens. The donor of one of the unsuitable platelet concentrates remembered when questioned, that he had eaten carrots and chocolate with hazelnuts during the evening before platelet donation. Two washed platelet concentrates were transfused without any problem. Furthermore, transfusions of nine red blood cell concentrates and one unit of virus-inactivated frozen pooled plasma were well tolerated. Patients should be asked for allergies previous to transfusions to be alert to allergic reactions in patients with a positive history of food or drug allergies. If premedication with antihistamines does not prevent severe allergic transfusion reactions, transfusion of washed platelet concentrates and of virus-inactivated frozen pooled plasma can be considered. © Georg Thieme Verlag KG Stuttgart · New York.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Lin, L.; Wiesehahn, G.P.; Morel, P.A.
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 R17more » 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.« less
Atar, Oliver D; Eisert, Christian; Pokov, Ilya; Serebruany, Victor L
2010-07-01
Sample fixation for storage and/or transportation represents an unsolved challenge for multicenter clinical trials assessing serial changes in platelet activity, or monitoring various antiplatelet regimens. Whole blood flow cytometry represents a major advance in defining platelet function, although special training and expensive equipment is required. We sought to determine how fixation with 2% paraformaldehyde (PFA), and storage of blood samples over 1 week affects the flow cytometry readings for both intact and thrombin-activating four major surface platelet receptors. Whole blood platelet expression of PECAM-1, P-selectin, PAR-1 inactive receptor (SPAN-12), and cleaved (WEDE-15) epitope was assessed immediately after blood draw, after staining with 2% PFA, and at day 1, 3, 5, and 7. The study was performed in 6 volunteers with multiple risk factors for vascular disease, not receiving any antiplatelet agents. Staining with PFA resulted in a slight decrease of fluorescence intensity, especially for PECAM-1, while antigen expression at day 1, 3 and 5 remains consistent, and highly reproducible. At day 7 there was a small but inconsistent trend towards diminished fluorescence intensity. The platelet data were consistent while validated with the isotype-matched irrelevant antibody. These data suggest that there is a 5 day window to perform final flow cytometry readings of whole blood PFA-fixed inactivated platelet samples. In contrast, thrombin activation cause gradual loss of flow cytometry signal, and cannot be recommended for long-term storage. This is critical logistic information for conducting multicenter platelet substudies within the framework of major clinical trials.
Du, Lily M; Nurden, Paquita; Nurden, Alan T; Nichols, Timothy C; Bellinger, Dwight A; Jensen, Eric S; Haberichter, Sandra L; Merricks, Elizabeth; Raymer, Robin A; Fang, Juan; Koukouritaki, Sevasti B; Jacobi, Paula M; Hawkins, Troy B; Cornetta, Kenneth; Shi, Qizhen; Wilcox, David A
2013-01-01
It is essential to improve therapies for controlling excessive bleeding in patients with haemorrhagic disorders. As activated blood platelets mediate the primary response to vascular injury, we hypothesize that storage of coagulation Factor VIII within platelets may provide a locally inducible treatment to maintain haemostasis for haemophilia A. Here we show that haematopoietic stem cell gene therapy can prevent the occurrence of severe bleeding episodes in dogs with haemophilia A for at least 2.5 years after transplantation. We employ a clinically relevant strategy based on a lentiviral vector encoding the ITGA2B gene promoter, which drives platelet-specific expression of human FVIII permitting storage and release of FVIII from activated platelets. One animal receives a hybrid molecule of FVIII fused to the von Willebrand Factor propeptide-D2 domain that traffics FVIII more effectively into α-granules. The absence of inhibitory antibodies to platelet-derived FVIII indicates that this approach may have benefit in patients who reject FVIII replacement therapies. Thus, platelet FVIII may provide effective long-term control of bleeding in patients with haemophilia A.
Du, Lily M.; Nurden, Paquita; Nurden, Alan T.; Nichols, Timothy C.; Bellinger, Dwight A.; Jensen, Eric S.; Haberichter, Sandra L.; Merricks, Elizabeth; Raymer, Robin A.; Fang, Juan; Koukouritaki, Sevasti B.; Jacobi, Paula M.; Hawkins, Troy B.; Cornetta, Kenneth; Shi, Qizhen; Wilcox, David A.
2013-01-01
It is essential to improve therapies for controlling excessive bleeding in patients with haemorrhagic disorders. As activated blood platelets mediate the primary response to vascular injury, we hypothesize that storage of coagulation Factor VIII within platelets may provide a locally inducible treatment to maintain haemostasis for haemophilia A. Here we show that haematopoietic stem cell gene therapy can prevent the occurrence of severe bleeding episodes in dogs with haemophilia A for at least 2.5 years after transplantation. We employ a clinically relevant strategy based on a lentiviral vector encoding the ITGA2B gene promoter, which drives platelet-specific expression of human FVIII permitting storage and release of FVIII from activated platelets. One animal receives a hybrid molecule of FVIII fused to the von Willebrand Factor propeptide-D2 domain that traffics FVIII more effectively into α-granules. The absence of inhibitory antibodies to platelet-derived FVIII indicates that this approach may have benefit in patients who reject FVIII replacement therapies. Thus, platelet FVIII may provide effective long-term control of bleeding in patients with haemophilia A. PMID:24253479
Proteome changes in platelets after pathogen inactivation--an interlaboratory consensus.
Prudent, Michel; D'Alessandro, Angelo; Cazenave, Jean-Pierre; Devine, Dana V; Gachet, Christian; Greinacher, Andreas; Lion, Niels; Schubert, Peter; Steil, Leif; Thiele, Thomas; Tissot, Jean-Daniel; Völker, Uwe; Zolla, Lello
2014-04-01
Pathogen inactivation (PI) of platelet concentrates (PCs) reduces the proliferation/replication of a large range of bacteria, viruses, and parasites as well as residual leucocytes. Pathogen-inactivated PCs were evaluated in various clinical trials showing their efficacy and safety. Today, there is some debate over the hemostatic activity of treated PCs as the overall survival of PI platelets seems to be somewhat reduced, and in vitro measurements have identified some alterations in platelet function. Although the specific lesions resulting from PI of PCs are still not fully understood, proteomic studies have revealed potential damages at the protein level. This review merges the key findings of the proteomic analyses of PCs treated by the Mirasol Pathogen Reduction Technology, the Intercept Blood System, and the Theraflex UV-C system, respectively, and discusses the potential impact on the biological functions of platelets. The complementarities of the applied proteomic approaches allow the coverage of a wide range of proteins and provide a comprehensive overview of PI-mediated protein damage. It emerges that there is a relatively weak impact of PI on the overall proteome of platelets. However, some data show that the different PI treatments lead to an acceleration of platelet storage lesions, which is in agreement with the current model of platelet storage lesion in pathogen-inactivated PCs. Overall, the impact of the PI treatment on the proteome appears to be different among the PI systems. Mirasol impacts adhesion and platelet shape change, whereas Intercept seems to impact proteins of intracellular platelet activation pathways. Theraflex influences platelet shape change and aggregation, but the data reported to date are limited. This information provides the basis to understand the impact of different PI on the molecular mechanisms of platelet function. Moreover, these data may serve as basis for future developments of PI technologies for PCs. Further studies should address the impact of both the PI and the storage duration on platelets in PCs because PI may enable the extension of the shelf life of PCs by reducing the bacterial contamination risk. Copyright © 2014 Elsevier Inc. All rights reserved.
Schubert, Peter; Johnson, Lacey; Marks, Denese C.; Devine, Dana V.
2018-01-01
Transfusions of platelets are an important cornerstone of medicine; however, recipients may be subject to risk of adverse events associated with the potential transmission of pathogens, especially bacteria. Pathogen inactivation (PI) technologies based on ultraviolet illumination have been developed in the last decades to mitigate this risk. This review discusses studies of platelet concentrates treated with the current generation of PI technologies to assess their impact on quality, PI capacity, safety, and clinical efficacy. Improved safety seems to come with the cost of reduced platelet functionality, and hence transfusion efficacy. In order to understand these negative impacts in more detail, several molecular analyses have identified signaling pathways linked to platelet function that are altered by PI. Because some of these biochemical alterations are similar to those seen arising in the context of routine platelet storage lesion development occurring during blood bank storage, we lack a complete picture of the contribution of PI treatment to impaired platelet functionality. A model generated using data from currently available publications places the signaling protein kinase p38 as a central player regulating a variety of mechanisms triggered in platelets by PI systems. PMID:29868586
Novel and unexpected clearance mechanisms for cold platelets
Rumjantseva, Viktoria; Hoffmeister, Karin M.
2015-01-01
Storage at room temperature is limited to 5 days because of the risk of bacterial growth and loss of platelet functionality. Platelet refrigeration remains impossible, because once chilled, platelets are rapidly removed from circulation. Chilling platelets (<4 h) clusters glycoprotein (GP) Ibα receptors, and β2 integrins on hepatic macrophages recognize clustered βGlcNAc residues leading to rapid clearance of acutely chilled platelets. Prolonged refrigeration increases the exposure of galactose residues such that, unexpectedly, hepatocytes remove platelets using their asialoglycoprotein receptors. Here we review current knowledge of the mechanisms of platelet removal, the existing knowledge of refrigerated platelet function, and methods to preserve platelet concentrates long-term for transfusion. PMID:19932055
Evaluation of four methods for platelet compatibility testing
DOE Office of Scientific and Technical Information (OSTI.GOV)
McFarland, J.G.; Aster, R.H.
1987-05-01
Four platelet compatibility assays were performed on serum and platelet or lymphocyte samples from 38 closely HLA-matched donor/recipient pairs involved in 55 single-donor platelet transfusions. The 22 patients studied were refractory to transfusions of pooled random-donor platelets. Of the four assays (platelet suspension immunofluorescence, PSIFT; /sup 51/Cr release; microlymphocytotoxicity; and a monoclonal anti-IgG assay, MAIA), the MAIA was most predictive of platelet transfusion outcome (predictability, 74% for one-hour posttransfusion platelet recovery and 76% for 24-hour recovery). The only other assay to reach statistical significance was the PSIFT (63% predictability for one-hour posttransfusion recovery). The degree of HLA compatibility between donormore » and recipient (exact matches v those utilizing cross-reactive associations) was unrelated to the ability of the MAIA to predict transfusion results. The MAIA may be capable of differentiating HLA antibodies, ABO antibodies, and platelet-specific antibodies responsible for failure of HLA-matched and selectively mismatched single-donor platelet transfusions.« less
Platelet Dynamics during Natural and Pharmacologically Induced Torpor and Forced Hypothermia
de Vrij, Edwin L.; Vogelaar, Pieter C.; Goris, Maaike; Houwertjes, Martin C.; Herwig, Annika; Dugbartey, George J.; Boerema, Ate S.; Strijkstra, Arjen M.; Bouma, Hjalmar R.; Henning, Robert H.
2014-01-01
Hibernation is an energy-conserving behavior in winter characterized by two phases: torpor and arousal. During torpor, markedly reduced metabolic activity results in inactivity and decreased body temperature. Arousal periods intersperse the torpor bouts and feature increased metabolism and euthermic body temperature. Alterations in physiological parameters, such as suppression of hemostasis, are thought to allow hibernators to survive periods of torpor and arousal without organ injury. While the state of torpor is potentially procoagulant, due to low blood flow, increased viscosity, immobility, hypoxia, and low body temperature, organ injury due to thromboembolism is absent. To investigate platelet dynamics during hibernation, we measured platelet count and function during and after natural torpor, pharmacologically induced torpor and forced hypothermia. Splenectomies were performed to unravel potential storage sites of platelets during torpor. Here we show that decreasing body temperature drives thrombocytopenia during torpor in hamster with maintained functionality of circulating platelets. Interestingly, hamster platelets during torpor do not express P-selectin, but expression is induced by treatment with ADP. Platelet count rapidly restores during arousal and rewarming. Platelet dynamics in hibernation are not affected by splenectomy before or during torpor. Reversible thrombocytopenia was also induced by forced hypothermia in both hibernating (hamster) and non-hibernating (rat and mouse) species without changing platelet function. Pharmacological torpor induced by injection of 5′-AMP in mice did not induce thrombocytopenia, possibly because 5′-AMP inhibits platelet function. The rapidness of changes in the numbers of circulating platelets, as well as marginal changes in immature platelet fractions upon arousal, strongly suggest that storage-and-release underlies the reversible thrombocytopenia during natural torpor. Possibly, margination of platelets, dependent on intrinsic platelet functionality, governs clearance of circulating platelets during torpor. PMID:24722364
[Platelet function in acute myeloid leukemia. II. Aggregation of isolated platelets].
Zawilska, K; Komarnicki, M; Mańka, B
1978-01-01
In 22 patients with acute myeloid leukaemia (17 cases of myeloblastic leukaemia, 4 cases of myelomonocytic leukaemia and 1 case of undifferentiated-cell leukaemia) platelets were isolated from the plasma by the method of Nicholls and Hampton as modified by Levy-Toledano by centrifugation in albumin gradient. The aim of platelet isolation was their "concentration" in cases of thrombocytopenia to values making possible aggregation tests, and platelet separation from the influence of plasma factors. Then aggregation of isolated platelets caused by ADP was studied. In 16 out of 22 patients a fall of aggregation was observed, with the mean values of aggregation rate and intensity were significantly lower. Parallelly done determinations of aggregating activity released from the platelets by thrombin showed lower values as compared with platelets from healthy subjects. In might be thought, in this connection, that the demonstrated reduction of isolated platelets is associated with a diminution of the nucleotide pool or disturbances of the platelet release reaction. The disturbances of the platelet release reaction. The disturbances of aggregation of isolated platelets and reduction of the aggregating activity were most pronounced in acute myelomonocytic leukaemia.
Amable, Paola Romina; Carias, Rosana Bizon Vieira; Teixeira, Marcus Vinicius Telles; da Cruz Pacheco, Italo; Corrêa do Amaral, Ronaldo José Farias; Granjeiro, José Mauro; Borojevic, Radovan
2013-06-07
Platelet-rich plasma (PRP) is nowadays widely applied in different clinical scenarios, such as orthopedics, ophthalmology and healing therapies, as a growth factor pool for improving tissue regeneration. Studies into its clinical efficiency are not conclusive and one of the main reasons for this is that different PRP preparations are used, eliciting different responses that cannot be compared. Platelet quantification and the growth factor content definition must be defined in order to understand molecular mechanisms behind PRP regenerative strength. Standardization of PRP preparations is thus urgently needed. PRP was prepared by centrifugation varying the relative centrifugal force, temperature, and time. Having quantified platelet recovery and yield, the two-step procedure that rendered the highest output was chosen and further analyzed. Cytokine content was determined in different fractions obtained throughout the whole centrifugation procedure. Our method showed reproducibility when applied to different blood donors. We recovered 46.9 to 69.5% of total initial platelets and the procedure resulted in a 5.4-fold to 7.3-fold increase in platelet concentration (1.4 × 10(6) to 1.9 × 10(6) platelets/μl). Platelets were highly purified, because only <0.3% from the initial red blood cells and leukocytes was present in the final PRP preparation. We also quantified growth factors, cytokines and chemokines secreted by the concentrated platelets after activation with calcium and calcium/thrombin. High concentrations of platelet-derived growth factor, endothelial growth factor and transforming growth factor (TGF) were secreted, together with the anti-inflammatory and proinflammatory cytokines interleukin (IL)-4, IL-8, IL-13, IL-17, tumor necrosis factor (TNF)-α and interferon (IFN)-α. No cytokines were secreted before platelet activation. TGF-β3 and IFNγ were not detected in any studied fraction. Clots obtained after platelet coagulation retained a high concentration of several growth factors, including platelet-derived growth factor and TGF. Our study resulted in a consistent PRP preparation method that yielded a cytokine and growth factor pool from different donors with high reproducibility. These findings support the use of PRP in therapies aiming for tissue regeneration, and its content characterization will allow us to understand and improve the clinical outcomes.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Ripley, S.; Wakefield, T.; Spaulding, S.
1985-05-01
In this investigation platelet deposition in polytetrafluroethylene (PTFE) thoracoabdominal grafts (TAC's) was evaluated using two different semi-quantitative techniques. Ten PTFE TAG's 6 mm in diameter and 30 cm in length were inserted into 10 mongrel dogs. One, 4 and 6 weeks after graft implantation the animals were injected with autologous In-111 platelets labelled by a modified Thakur technique. Platelet imaging in grafts was performed 48 hrs after injection. Blood pool was determined by Tc99m labelled RBC's (in vivo/in vitro technique). Semi-quantitative analysis was performed by subdividing the imaged graft into three major regions and selecting a reference region from eithermore » the native aorta or common iliac artery. Excess platelet deposition was determined by two methods: 1) the ratio of In-111 counts in the graft ROI''s to the reference region and 2) the percent In-111 excess using the Tc99m blood pool subtraction technique (TBPST). Animals were sacrificed 7 weeks after implantation and radioactivity in the excised grafts was determined using a well counter. A positive correlation was found to exist between the In-111 ratio percent analysis (IRPA) and the direct gamma counting (DCC) for all three segments of the prosthetic graft. Correlation coefficients for the thorax, midsegment and abdominal segments were 0.80, 0.73 and 0.48 respectivly. There was no correlation between TBPST and DGC. Using the IRPA technique the thrombogenicity of TAG's can be routinely assessed and is clinically applicable for patient use. TBPST should probably be limited to the extremities to avoid error due to free Tc99m counts from kidneys and ureters.« less
Improved BC method of Compomat G4 for expression of BCs twice from whole blood in top and top bags.
Deng, Xiao-Yan; Wu, Xiao-Man; Zhao, Yang; Luo, Hong; Jia, Hong-Yun; Wang, Zhong-Ying; He, Bo; Wang, Chuan-Xi
2011-05-01
The aim of this paper was to evaluate an improved buffy coat (BC) method of Compomat G4 for automated expression of BCs twice from whole blood (WB) in top and top (T&T) bags. WB was separated using hard spin centrifugation (2,988g, 10 min) into layers of blood components by specific gravity, and different components were subsequently expressed into satellite bags in the T&T system using the manual BC method, the conventional BC method of G4, and our improved BC method of G4. In the improved BC method, an accessorial device we have named a 'gravity press' was designed and installed on the top flat of G4 to produce gravitational pressure on the plasma bag so as to exclude air and some of plasma to the upper compartment of the slide after BCs were expressed for the first time. The residual BCs in the upper compartment were expressed a second time by extending the upper press once more. All of the pooled BCs were centrifuged by soft spin (402g, 10 min) and upper platelet-rich supernatant was manually expressed into a platelet container by the plasma extractor. In vitro studies of blood components and pooled platelet concentrates (PCs) revealed no significant differences in BC blood components and platelet recovery of pooled platelets (61 ± 9 vs. 60 ± 7%, n = 12, p > 0.05) between the improved BC method and the conventional BC method; all components met our specifications for blood products. We suggest that the new BC method for use of T&T bags may improve the collection of BCs.
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.
Messenger RNA profiling of human platelets by microarray hybridization.
Bugert, Peter; Dugrillon, Alex; Günaydin, Ayse; Eichler, Hermann; Klüter, Harald
2003-10-01
Platelets are generally believed to be inactive in terms of de novo protein synthesis. On the other hand, the presence of ribosomes and mRNA molecules is well established. Many studies have used reverse transcriptase (RT) -PCR for detection of gene transcripts in platelets. As RT-PCR is a very sensitive method, any leukocyte contamination of platelet preparations can lead to false results. We performed three filtration procedures to minimize leukocyte contamination of pooled buffy-coat platelet concentrates prior to RNA isolation. Furthermore, by applying a genomic PCR approach with 50 amplification cycles we demonstrated that nucleated cells were not detectable. Microarray hybridization was used to analyze 9,850 individual human genes in RNA from purified platelets. In total we identified 1,526 (15.5%) positive genes. The data were confirmed in six individual experiments each performed on a PC pooled from four individual blood donations. Genes specific for nucleated blood cells such as CD4, CD83 and others were negative and verified the purity of PC. Overrepresentation of positive genes was found in the functional categories of glycoproteins/integrins (22.6% vs. 15.5%, p=0.029) and receptors (20.7% vs. 15.5%, p<0.001). Gene transcripts encoding RANTES, GRO-alpha, MIP-1alpha, MIP-1beta, and others were found at high levels of signal intensity and confirmed literature data. This work provides a mRNA profile of human platelets and a complete list of results can be downloaded from the website of our institute www.ma.uni-heidelberg.de/inst/iti/plt_array.xls. The knowledge about gene transcripts may have an impact on the characterization of novel proteins and their functions in platelets.
Storage of red blood cells in a novel polyolefin blood container: a pilot in vitro study.
Gulliksson, H; Meinke, S; Ravizza, A; Larsson, L; Höglund, P
2017-01-01
The present general plasticizer di-2-ethylhexyl-phthalate in polyvinylchloride (PVC) blood bags is only physically dispersed in PVC and will therefore leach into blood components. The objective of this study was to perform a first preliminary red blood cell (RBC) storage evaluation in a new blood bag manufactured of polyolefin without any inclusion of potentially migrating substances. This is a RBC storage study for 42 days. Blood collection was performed in a polyolefin-based PVC-free blood bag. RBCs were prepared within 8 h. Two different RBC additive solutions were used, either PAGGS-M or PAGGG-M. We weekly measured pH, K + , glucose, lactate, haemolysis, red cell ATP and 2,3-DPG. RBC storage in PAGGS-M resulted in high haemolysis levels already after 21 days, exceeding the European maximum limit of 0·8%, and low ATP levels by the end of the storage period. With PAGGG-M, haemolysis exceeded 0·8% after 28 days of storage. For additional parameters, the results were comparable to those of previous studies in conventional blood bags. This is a first preliminary study of RBC storage in a new type of blood bags. PAGGG-M gave encouraging results except for its inability to prevent increased haemolysis. There will be room for further development of RBC additive solutions to address the haemolysis problems. Plasma should also be tested regarding the stability of coagulation and activation pathway variables. There may also be a potential for future use of the bag for preparation of pooled buffy-coat-derived platelets. © 2016 International Society of Blood Transfusion.
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.
The clearance mechanism of chilled blood platelets.
Hoffmeister, Karin M; Felbinger, Thomas W; Falet, Hervé; Denis, Cécile V; Bergmeier, Wolfgang; Mayadas, Tanya N; von Andrian, Ulrich H; Wagner, Denisa D; Stossel, Thomas P; Hartwig, John H
2003-01-10
Platelet transfusion is a very common lifesaving medical procedure. Not widely known is the fact that platelets, unlike other blood cells, rapidly leave the circulation if refrigerated prior to transfusion. This peculiarity requires blood services to store platelets at room temperature, limiting platelet supplies for clinical needs. Here, we describe the mechanism of this clearance system, a longstanding mystery. Chilling platelets clusters their von Willebrand (vWf) receptors, eliciting recognition of mouse and human platelets by hepatic macrophage complement type 3 (CR3) receptors. CR3-expressing but not CR3-deficient mice exposed to cold rapidly decrease platelet counts. Cooling primes platelets for activation. We propose that platelets are thermosensors, primed at peripheral sites where most injuries occurred throughout evolution. Clearance prevents pathologic thrombosis by primed platelets. Chilled platelets bind vWf and function normally in vitro and ex vivo after transfusion into CR3-deficient mice. Therefore, GPIb modification might permit cold platelet storage.
Schubert, Peter; Culibrk, Brankica; Karwal, Simrath; Serrano, Katherine; Levin, Elena; Bu, Daniel; Bhakta, Varsha; Sheffield, William P; Goodrich, Raymond P; Devine, Dana V
2015-04-01
Pathogen inactivation (PI) technologies are currently licensed for use with platelet (PLT) and plasma components. Treatment of whole blood (WB) would be of benefit to the blood banking community by saving time and costs compared to individual component treatment. However, no paired, pool-and-split study directly assessing the impact of WB PI on the subsequently produced components has yet been reported. In a "pool-and-split" study, WB either was treated with riboflavin and ultraviolet (UV) light or was kept untreated as control. The buffy coat (BC) method produced plasma, PLT, and red blood cell (RBC) components. PLT units arising from the untreated WB study arm were treated with riboflavin and UV light on day of production and compared to PLT concentrates (PCs) produced from the treated WB units. A panel of common in vitro variables for the three types of components was used to monitor quality throughout their respective storage periods. PCs derived from the WB PI treatment were of significantly better quality than treated PLT components for most variables. RBCs produced from the WB treatment deteriorated earlier during storage than untreated units. Plasma components showed a 3% to 44% loss in activity for several clotting factors. Treatment of WB with riboflavin and UV before production of components by the BC method shows a negative impact on all three blood components. PLT units produced from PI-treated WB exhibited less damage compared to PLT component treatment. © 2014 AABB.
Seghatchian, Jerard; Amiral, Jean
2016-08-01
Blood cells generate heterogeneous populations of vesicles that are delivered, as small-specialized packages of highly active cell fragments in blood circulation, having almost similar functional activities, as the mother cells. These so called extracellular vesicles are the essential part of an energy-dependent natural apoptotic process; hence their beneficial and harmful biological functions cannot be ignored. Evidence is accumulating, that cellular derived vesicles, originate from all viable cells including: megakaryocytes, platelets, red blood cells, white blood cells and endothelial cells, the highest in proportions from platelets. Shedding can also be triggered by pathological activation of inflammatory processes and activation of coagulation or complement pathways, or even by shear stress in the circulation. Structurally, so called MV/EV appear to be, sometimes inside-out and sometimes outside-in cell fragments having a bilayered phospholipid structure exposing coagulant-active phosphatidylserine, expressing various membrane receptors, and they serve as cell-to-cell shuttles for bioactive molecules such as lipids, growth factors, microRNAs, and mitochondria. Ex vivo processing of blood into its components, embodying centrifugation, processing by various apheresis procedures, leukoreduction, pathogen reduction, and finally storage in different media and different types of blood bags, also have major impacts on the generation and retention of MV content. These artificially generated small, but highly liable packages, together with the original pool of MVs collected from the donor, do exhibit differing biological activities, and are not inert elements and should be considered as a parameter of blood safety in haemovigilance programmes. Harmonization and consensus in sampling protocols, sample handling, processing, and assessment methods, in particular converting to full automation, are needed to achieve consensual interpretations. This review focuses on some of our past personal studies on the role of MV/EV focusing on characterization of platelet storage lesion and platelet therapy that shows the highest transfusion hazards [up to 25%], and loss of 25% platelet efficacy after various leukoreduction and validated platelet pathogen reduction treatments. The planned paths for the future of EV/MV involvement in immunological and viral/ non-viral transfusion hazards are also discussed. Whilst considerable advances made on the characterization of EV/MV, but disparity still exists between various surrogate markers, showing some subtle differences in the levels of MV/ EV & BRMs in platelet preparations, and the clinical outcome showing platelets derived by all current technologies are equivalents in vivo. One possible reason for such a disparity may be relatedto the fact that MVs, being the end products of apoptotic cells, have little specificity and clear rapidly from circulation [<6 h in thrombocytopoenia]. This makes their clinical usefulness rather short lived. The recent findings that pegylating smaller subsets of EV increases its circulatory life from <15 minutes to approximately about one hour is highly promising, in particular, for drug delivery on specific sides. Hence a promising clinical utility of EV/MV continues, as a journey without end, indeed. This manuscript is based mainly on the selected key readings listed below. Copyright © 2016. Published by Elsevier Ltd.
Arf6 controls platelet spreading and clot retraction via integrin αIIbβ3 trafficking
Huang, Yunjie; Joshi, Smita; Xiang, Binggang; Kanaho, Yasunori; Li, Zhenyu; Bouchard, Beth A.; Moncman, Carole L.
2016-01-01
Platelet and megakaryocyte endocytosis is important for loading certain granule cargo (ie, fibrinogen [Fg] and vascular endothelial growth factor); however, the mechanisms of platelet endocytosis and its functional acute effects are understudied. Adenosine 5'-diphosphate–ribosylation factor 6 (Arf6) is a small guanosine triphosphate–binding protein that regulates endocytic trafficking, especially of integrins. To study platelet endocytosis, we generated platelet-specific Arf6 knockout (KO) mice. Arf6 KO platelets had less associated Fg suggesting that Arf6 affects αIIbβ3-mediated Fg uptake and/or storage. Other cargo was unaffected. To measure Fg uptake, mice were injected with biotinylated- or fluorescein isothiocyanate (FITC)–labeled Fg. Platelets from the injected Arf6 KO mice showed lower accumulation of tagged Fg, suggesting an uptake defect. Ex vivo, Arf6 KO platelets were also defective in FITC-Fg uptake and storage. Immunofluorescence analysis showed initial trafficking of FITC-Fg to a Rab4-positive compartment followed by colocalization with Rab11-positive structures, suggesting that platelets contain and use both early and recycling endosomes. Resting and activated αIIbβ3 levels, as measured by flow cytometry, were unchanged; yet, Arf6 KO platelets exhibited enhanced spreading on Fg and faster clot retraction. This was not the result of alterations in αIIbβ3 signaling, because myosin light-chain phosphorylation and Rac1/RhoA activation were unaffected. Consistent with the enhanced clot retraction and spreading, Arf6 KO mice showed no deficits in tail bleeding or FeCl3-induced carotid injury assays. Our studies present the first mouse model for defining the functions of platelet endocytosis and suggest that altered integrin trafficking may affect the efficacy of platelet function. PMID:26738539
Arf6 controls platelet spreading and clot retraction via integrin αIIbβ3 trafficking.
Huang, Yunjie; Joshi, Smita; Xiang, Binggang; Kanaho, Yasunori; Li, Zhenyu; Bouchard, Beth A; Moncman, Carole L; Whiteheart, Sidney W
2016-03-17
Platelet and megakaryocyte endocytosis is important for loading certain granule cargo (ie, fibrinogen [Fg] and vascular endothelial growth factor); however, the mechanisms of platelet endocytosis and its functional acute effects are understudied. Adenosine 5'-diphosphate-ribosylation factor 6 (Arf6) is a small guanosine triphosphate-binding protein that regulates endocytic trafficking, especially of integrins. To study platelet endocytosis, we generated platelet-specific Arf6 knockout (KO) mice. Arf6 KO platelets had less associated Fg suggesting that Arf6 affects αIIbβ3-mediated Fg uptake and/or storage. Other cargo was unaffected. To measure Fg uptake, mice were injected with biotinylated- or fluorescein isothiocyanate (FITC)-labeled Fg. Platelets from the injected Arf6 KO mice showed lower accumulation of tagged Fg, suggesting an uptake defect. Ex vivo, Arf6 KO platelets were also defective in FITC-Fg uptake and storage. Immunofluorescence analysis showed initial trafficking of FITC-Fg to a Rab4-positive compartment followed by colocalization with Rab11-positive structures, suggesting that platelets contain and use both early and recycling endosomes. Resting and activated αIIbβ3 levels, as measured by flow cytometry, were unchanged; yet, Arf6 KO platelets exhibited enhanced spreading on Fg and faster clot retraction. This was not the result of alterations in αIIbβ3 signaling, because myosin light-chain phosphorylation and Rac1/RhoA activation were unaffected. Consistent with the enhanced clot retraction and spreading, Arf6 KO mice showed no deficits in tail bleeding or FeCl3-induced carotid injury assays. Our studies present the first mouse model for defining the functions of platelet endocytosis and suggest that altered integrin trafficking may affect the efficacy of platelet function. © 2016 by The American Society of Hematology.
Fekete, Natalie; Gadelorge, Mélanie; Fürst, Daniel; Maurer, Caroline; Dausend, Julia; Fleury-Cappellesso, Sandrine; Mailänder, Volker; Lotfi, Ramin; Ignatius, Anita; Sensebé, Luc; Bourin, Philippe; Schrezenmeier, Hubert; Rojewski, Markus Thomas
2012-01-01
Background aims The clinical use of human mesenchymal stromal cells (MSC) requires ex vivo expansion in media containing supplements such as fetal bovine serum or, alternatively, human platelet lysate (PL). Methods Platelet concentrates were frozen, quarantine stored, thawed and sterile filtered to obtain PL. PL content and its effect on fibroblast-colony-forming unit (CFU-F) formation, MSC proliferation and large-scale expansion were studied. Results PL contained high levels of basic fibroblast growth factor (bFGF), soluble CD40L (sCD40L), vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), platelet-derived growth factor AA (PDGF-AA), platelet-derived growth factor AB/BB (PDGF-AB/BB), chemokine (C-C) ligand 5 (CCL5; RANTES) transforming growth factor-β1 (TGF-β1) and chemokine (C-X-C) ligand 1/2/3 (GRO), with low batch-to-batch variability, and most were stable for up to 14 days. Inhibition of PDGF-BB and bFGF decreased MSC proliferation by about 20% and 50%, respectively. The strongest inhibition (about 75%) was observed with a combination of anti-bFGF + anti-PDGF-BB and anti-bFGF + anti-TGF-β1 + anti-PDGF-BB. Interestingly, various combinations of recombinant PDGF-BB, bFGF and TGF-β1 were not sufficient to promote cell proliferation. PL from whole blood-derived pooled platelet concentrates and apheresis platelet concentrates did not differ significantly in their growth-promoting activity on MSC. Conclusions PL enhances MSC proliferation and can be regarded as a safe tool for MSC expansion for clinical purposes. \\in particular, PDGF-BB and bFGF are essential components for the growth-promoting effect of PL, but are not sufficient for MSC proliferation. PMID:22296115
1991-01-31
that a short incubation of platelets with 0.1% (final conc.) KMnO4 seems optimal, but a protective agent such as trehalose , sucrose, or albumin must...yields were improved also by inclusion of albumin during platelet washing and trehalose during the lyophilization step (74% recovery under best... trehalose is advantageous in this protocol has yet to be decided. Vesiculation of platelet membranes is still evident by flow cytometry in the
Blood platelets: computerized morphometry applied on optical images
NASA Astrophysics Data System (ADS)
Korobova, Farida V.; Ivanova, Tatyana V.; Gusev, Alexander A.; Shmarov, Dmitry A.; Kozinets, Gennady I.
2000-11-01
The new technology of computerized morphometric image analysis of platelets on blood smears was developed. In a basis of the device is included analysis of cytophotometric and morphometric parameters of platelets. Geometrical and optical parameters of platelets on 35 donors, platelet concentrates and 15 patients with haemorrhagic thrombocythaemia were investigated, average meanings for the area, diameter, its logarithms and optical density of platelets in norm were received. Distribution of the areas, diameters and optical densities of platelets of patients with haemorrhagic thrombocythaemia differed from those at the healthy people. After a course of treatment these meanings came nearer to normal. The important characteristics of platelets in platelet concentrates after three days of storage were in limits of normal meanings, but differed from those in whole blood platelets. Obtained data allow to enter the quantitative standards into investigation of platelets of the healthy people and at various alteration of thrombocytopoieses.
Dual roles for hepatic lectin receptors in the clearance of chilled platelets.
Rumjantseva, Viktoria; Grewal, Prabhjit K; Wandall, Hans H; Josefsson, Emma C; Sørensen, Anne Louise; Larson, Göran; Marth, Jamey D; Hartwig, John H; Hoffmeister, Karin M
2009-11-01
Rapid chilling causes glycoprotein-Ib (GPIb) receptors to cluster on blood platelets. Hepatic macrophage beta(2) integrin binding to beta-N-acetylglucosamine (beta-GlcNAc) residues in the clusters leads to rapid clearance of acutely chilled platelets after transfusion. Although capping the beta-GlcNAc moieties by galactosylation prevents clearance of short-term-cooled platelets, this strategy is ineffective after prolonged refrigeration. We report here that prolonged refrigeration increased the density and concentration of exposed galactose residues on platelets such that hepatocytes, through Ashwell-Morell receptor binding, become increasingly involved in platelet removal. Macrophages rapidly removed a large fraction of transfused platelets independent of their storage conditions. With prolonged platelet chilling, hepatocyte-dependent clearance further diminishes platelet recovery and survival after transfusion. Inhibition of chilled platelet clearance by both beta(2) integrin and Ashwell-Morell receptors may afford a potentially simple method for storing platelets in the cold.
Pitman, John P.; Basavaraju, Sridhar V.; Shiraishi, Ray W.; Wilkinson, Robert; von Finckenstein, Bjorn; Lowrance, David W.; Marfin, Anthony A.; Postma, Maarten; Mataranyika, Mary; Sibinga, Cees Th. Smit
2015-01-01
BACKGROUND Few African countries separate blood donations into components; however, demand for platelets (PLTs) is increasing as regional capacity to treat causes of thrombocytopenia, including chemotherapy, increases. Namibia introduced single-donor apheresis PLT collections in 2007 to increase PLT availability while reducing exposure to multiple donors via pooling. This study describes the impact this transition had on PLT availability and safety in Namibia. STUDY DESIGN AND METHODS Annual national blood collections and PLT units issued data were extracted from a database maintained by the Blood Transfusion Service of Namibia (NAMBTS). Production costs and unit prices were analyzed. RESULTS In 2006, NAMBTS issued 771 single and pooled PLT doses from 3054 whole blood (WB) donations (drawn from 18,422 WB donations). In 2007, NAMBTS issued 486 single and pooled PLT doses from 1477 WB donations (drawn from 18,309 WB donations) and 131 single-donor PLT doses. By 2011, NAMBTS issued 837 single-donor PLT doses per year, 99.1% of all PLT units. Of 5761 WB donations from which PLTs were made in 2006 to 2011, a total of 20 (0.35%) were from donors with confirmed test results for human immunodeficiency virus or other transfusion-transmissible infections (TTIs). Of 2315 single-donor apheresis donations between 2007 and 2011, none of the 663 donors had a confirmed positive result for any pathogen. As apheresis replaced WB-derived PLTs, apheresis production costs dropped by a mean of 8.2% per year, while pooled PLT costs increased by an annual mean of 21.5%. Unit prices paid for apheresis- and WB-derived PLTs increased by 9 and 7.4% per year on average, respectively. CONCLUSION Namibia’s PLT transition shows that collections from repeat apheresis donors can reduce TTI risk and production costs. PMID:25727921
NASA Technical Reports Server (NTRS)
Otsuka, K.; Watanabe, M.; Yue, Q.; McCarron, D. A.; Hatton, D.
1997-01-01
Spontaneously hypertensive rats (SHR) are known to be blood pressure sensitive to dietary calcium. The effects of dietary calcium on platelet aggregation and intracellular Ca2+ mobilization were assessed by turbidimetric methods and fura-2 methods, respectively, in washed platelets of SHR. Ca2+ ATPase activity was examined in aortic membrane fractions. Six weeks of dietary calcium supplementation attenuated the increase of systolic blood pressure (SBP 199 +/- 16 v 170 +/- 9 mm Hg, P < .001) and thrombin-induced platelet aggregation (84.5 +/- 3.7 v 73.7 +/- 7.4%, P < .004) at 9 weeks of age. The ionomycin-induced intracellular calcium ([Ca2+]i) peak in the absence of external Ca2+, which reflects [Ca2+]i storage size, and thrombin-evoked [Ca2+]i release from [Ca2+]i storage were decreased by 2.0% Ca diet (472 +/- 55 v 370 +/- 23 nmol/L, P < .001, 339 +/- 29 v 278 +/- 33 nmol/L, P < .002). In addition, SBP was positively correlated with platelet aggregation (r = 0.703, P = .0088), thrombin-evoked [Ca2+]i (r = 0.739, P = .0044), and ionomycin-induced [Ca2+]i (r = 0.591, P = .0415), respectively. However, there was no significant effect of dietary calcium on Ca2+-ATPase activity in aortic membranes. These results suggest that dietary calcium supplementation had a beneficial effect on platelets of SHR by attenuating [Ca2+]i mobilization from [Ca2+]i storage. The hypotensive effect of dietary calcium might be associated with attenuated [Ca2+]i mobilization in SHR.
Allocation to carbon storage pools in Norway spruce saplings under drought and low CO2.
Hartmann, Henrik; McDowell, Nate G; Trumbore, Susan
2015-03-01
Non-structural carbohydrates (NSCs) are critical to maintain plant metabolism under stressful environmental conditions, but we do not fully understand how NSC allocation and utilization from storage varies with stress. While it has become established that storage allocation is unlikely to be a mere overflow process, very little empirical evidence has been produced to support this view, at least not for trees. Here we present the results of an intensively monitored experimental manipulation of whole-tree carbon (C) balance (young Picea abies (L.) H Karst.) using reduced atmospheric [CO2] and drought to reduce C sources. We measured specific C storage pools (glucose, fructose, sucrose, starch) over 21 weeks and converted concentration measurement into fluxes into and out of the storage pool. Continuous labeling ((13)C) allowed us to track C allocation to biomass and non-structural C pools. Net C fluxes into the storage pool occurred mainly when the C balance was positive. Storage pools increased during periods of positive C gain and were reduced under negative C gain. (13)C data showed that C was allocated to storage pools independent of the net flux and even under severe C limitation. Allocation to below-ground tissues was strongest in control trees followed by trees experiencing drought followed by those grown under low [CO2]. Our data suggest that NSC storage has, under the conditions of our experimental manipulation (e.g., strong progressive drought, no above-ground growth), a high allocation priority and cannot be considered an overflow process. While these results also suggest active storage allocation, definitive proof of active plant control of storage in woody plants requires studies involving molecular tools. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.
Antimicrobial effect of platelet-rich plasma and platelet-rich fibrin.
Badade, Pallavi S; Mahale, Swapna A; Panjwani, Alisha A; Vaidya, Prutha D; Warang, Ayushya D
2016-01-01
Platelet concentrates have been extensively used in a variety of medical fields to promote soft- and hard-tissue regeneration. The significance behind their use lies in the abundance of growth factors (GFs) in platelets α-granules that promote wound healing. Other than releasing a pool of GFs upon activation, platelets also have many features that indicate their role in the anti-infective host defense. The aim of this study is to evaluate the antimicrobial activities of platelet-rich plasma (PRP) and platelet-rich fibrin (PRF) against periodontal disease-associated bacteria. Blood samples were obtained from ten adult male patients. PRP and PRF were procured using centrifugation. The antimicrobial activity of PRP and PRF was evaluated by microbial culturing using bacterial strains of Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans. P. gingivalis and A. actinomycetemcomitans were inhibited by PRP but not by PRF. PRP is a potentially useful substance in the fight against periodontal pathogens. This might represent a valuable property in adjunct to the enhancement of tissue regeneration.
van der Wal, Dianne E.; Gitz, Eelo; Du, Vivian X.; Lo, Kimberly S.L.; Koekman, Cornelis A.; Versteeg, Sabine; Akkerman, Jan Willem N.
2012-01-01
Background Cold storage of platelets reduces bacterial growth and preserves their hemostatic properties better than current procedures do. However, storage at 0°C induces [14-3-3ζ-glycoprotein Ibα] association, 14-3-3ζ release from phospho-Bad, Bad activation and apoptosis. Design and Methods We investigated whether arachidonic acid, which also binds 14-3-3ζ, contributes to coldinduced apoptosis. Results Cold storage activated P38-mitogen-activated protein kinase and released arachidonic acid, which accumulated due to cold inactivation of cyclooxygenase-1/thromboxane synthase. Accumulated arachidonic acid released 14-3-3ζ from phospho-Bad and decreased the mitochondrial membrane potential, which are steps in the induction of apoptosis. Addition of arachidonic acid did the same and its depletion made platelets resistant to cold-induced apoptosis. Incubation with biotin-arachidonic acid revealed formation of an [arachidonic acid-14-3-3ζ-glycoprotein Ibα] complex. Indomethacin promoted complex formation by accumulating arachidonic acid and released 14-3-3ζ from cyclo-oxygenase-1. Arachidonic acid depletion prevented the cold-induced reduction of platelet survival in mice. Conclusions We conclude that cold storage induced apoptosis through an [arachidonic acid-14-3-3ζ-glycoprotein Ibα] complex, which released 14-3-3ζ from Bad in an arachidonic acid-dependent manner. Although arachidonic acid depletion reduced agonist-induced thromboxane A2 formation and aggregation, arachidonic acid repletion restored these functions, opening ways to reduce apoptosis during storage without compromising hemostatic functions post-transfusion. PMID:22371179
Qadri, Syed M; Chen, Deborah; Schubert, Peter; Perruzza, Darian L; Bhakta, Varsha; Devine, Dana V; Sheffield, William P
2017-03-01
Pathogen reduction treatment using riboflavin and ultraviolet light illumination (Mirasol) effectively reduces the risk of transfusion-transmitted infections. This treatment is currently licensed for only platelets and plasma products, while its application to whole blood (WB) to generate pathogen-inactivated red blood cells (RBCs) is under development. RBC storage lesion, constituting numerous morphologic and biochemical changes, influences RBC quality and limits shelf life. Stored RBCs further show enhanced susceptibility to RBC programmed cell death (eryptosis) characterized by increased cytosolic Ca 2+ -provoked membrane phosphatidylserine (PS) externalization. Using a "pool-and-split" approach, we examined multiple variables of RBC storage lesion and eryptosis in RBC units, derived from Mirasol-treated or untreated WB, after 4 to 42 days of storage, under blood bank conditions. In comparison to untreated RBC units, Mirasol treatment significantly altered membrane microvesiculation, supernatant hemoglobin, osmotic fragility, and intracellular adenosine triphosphate levels but did not influence membrane CD47 expression and 2,3-diphosphoglycerate levels. Mirasol-treated RBCs showed significantly higher PS exposure after 42, but not after not more than 21, days of storage, which was accompanied by enhanced cytosolic Ca 2+ activity, ceramide abundance, and oxidative stress, but not p38 kinase activation. Mirasol treatment significantly augmented PS exposure, Ca 2+ entry, and protein kinase C activation after energy depletion, a pathophysiologic cell stressor. Mirasol-treated RBCs were, however, more resistant to cell shrinkage. Prolonged storage of Mirasol-treated RBCs significantly increases the proportion of eryptotic RBCs, while even short-term storage enhances the susceptibility of RBCs to stress-induced eryptosis, which could reduce posttransfusion RBC recovery in patients. © 2016 AABB.
Jansen, A J Gerard; Josefsson, Emma C; Rumjantseva, Viktoria; Liu, Qiyong Peter; Falet, Hervé; Bergmeier, Wolfgang; Cifuni, Stephen M; Sackstein, Robert; von Andrian, Ulrich H; Wagner, Denisa D; Hartwig, John H; Hoffmeister, Karin M
2012-02-02
When refrigerated platelets are rewarmed, they secrete active sialidases, including the lysosomal sialidase Neu1, and express surface Neu3 that remove sialic acid from platelet von Willebrand factor receptor (VWFR), specifically the GPIbα subunit. The recovery and circulation of refrigerated platelets is greatly improved by storage in the presence of inhibitors of sialidases. Desialylated VWFR is also a target for metalloproteinases (MPs), because GPIbα and GPV are cleaved from the surface of refrigerated platelets. Receptor shedding is inhibited by the MP inhibitor GM6001 and does not occur in Adam17(ΔZn/ΔZn) platelets expressing inactive ADAM17. Critically, desialylation in the absence of MP-mediated receptor shedding is sufficient to cause the rapid clearance of platelets from circulation. Desialylation of platelet VWFR therefore triggers platelet clearance and primes GPIbα and GPV for MP-dependent cleavage.
Jansen, A. J. Gerard; Josefsson, Emma C.; Rumjantseva, Viktoria; Liu, Qiyong Peter; Falet, Hervé; Bergmeier, Wolfgang; Cifuni, Stephen M.; Sackstein, Robert; von Andrian, Ulrich H.; Wagner, Denisa D.; Hartwig, John H.
2012-01-01
When refrigerated platelets are rewarmed, they secrete active sialidases, including the lysosomal sialidase Neu1, and express surface Neu3 that remove sialic acid from platelet von Willebrand factor receptor (VWFR), specifically the GPIbα subunit. The recovery and circulation of refrigerated platelets is greatly improved by storage in the presence of inhibitors of sialidases. Desialylated VWFR is also a target for metalloproteinases (MPs), because GPIbα and GPV are cleaved from the surface of refrigerated platelets. Receptor shedding is inhibited by the MP inhibitor GM6001 and does not occur in Adam17ΔZn/ΔZn platelets expressing inactive ADAM17. Critically, desialylation in the absence of MP-mediated receptor shedding is sufficient to cause the rapid clearance of platelets from circulation. Desialylation of platelet VWFR therefore triggers platelet clearance and primes GPIbα and GPV for MP-dependent cleavage. PMID:22101895
Functional factor XIII-A is exposed on the stimulated platelet surface
Mitchell, Joanne L.; Lionikiene, Ausra S.; Fraser, Steven R.; Whyte, Claire S.; Booth, Nuala A.
2014-01-01
Factor XIII (FXIII) stabilizes thrombi against fibrinolysis by cross-linking α2-antiplasmin (α2AP) to fibrin. Cellular FXIII (FXIII-A) is abundant in platelets, but the extracellular functions of this pool are unclear because it is not released by classical secretion mechanisms. We examined the function of platelet FXIII-A using Chandler model thrombi formed from FXIII-depleted plasma. Platelets stabilized FXIII-depleted thrombi in a transglutaminase-dependent manner. FXIII-A activity on activated platelets was unstable and was rapidly lost over 1 hour. Inhibiting platelet activation abrogated the ability of platelets to stabilize thrombi. Incorporating a neutralizing antibody to α2AP into FXIII-depleted thrombi revealed that the stabilizing effect of platelet FXIII-A on lysis was α2AP dependent. Platelet FXIII-A activity and antigen were associated with the cytoplasm and membrane fraction of unstimulated platelets, and these fractions were functional in stabilizing FXIII-depleted thrombi against lysis. Fluorescence confocal microscopy and flow cytometry revealed exposure of FXIII-A on activated membranes, with maximal signal detected with thrombin and collagen stimulation. FXIII-A was evident in protruding caps on the surface of phosphatidylserine-positive platelets. Our data show a functional role for platelet FXIII-A through exposure on the activated platelet membrane where it exerts antifibrinolytic function by cross-linking α2AP to fibrin. PMID:25331118
Rank, A; Nieuwland, R; Liebhardt, S; Iberer, M; Grützner, S; Toth, B; Pihusch, R
2011-02-01
Microparticles (MP) are membrane vesicles with thrombogenic and immunomodulatory properties. We determined MP subgroups from resting platelets, activated platelets and endothelial cells in donors and apheresis platelet concentrates (PC). MP were double stained with annexin V and CD61 (platelet-derived MP; PMP), P-selectin or CD63 (MP from activated platelets) and CD144 plus E-selectin (endothelial cell-derived MP; EMP) and detected by flow cytometry in platelet donors (n=36) and apheresis PC (n=11; Trima™). PC contained MP, mainly from resting platelets [93% (90-95)], and minor fractions of PMP from activated platelets [P-selectin(+) or CD63(+); 4·8% (3·2-7·7) and 2·6% (2·0-4·0)]. Compared to donors, levels of annexin V+ MP, PMP, P-selectin(+) and CD63(+) MP were 1·7-, 2·3-, 8·6- and 3·1-fold higher in PC (all P<0·05). During storage (1-5 days), levels of annexin V+ MP and PMP did not increase, although small increases in the fraction of P-selectin(+) or CD63(+) MP occurred (both P<0·05). PC also contained EMP, which were 2·6- to 3·7-fold enriched in PC compared to donors (P<0·05). Transfusion of apheresis PC also results in transfusion of HLA-carrying PMP and EMP. This might counteract the aim of reducing transfused HLA load by leucodepletion. The increases in PMP exposing P-selectin or CD63 reflect mild platelet activation during storage. We conclude that in leucodepleted platelet apheresis using fluidized particle bed technology, MP are harvested mainly from the donor by apheresis. Improvement in apheresis technology might reduce MP load. © 2010 The Author(s). Vox Sanguinis © 2010 International Society of Blood Transfusion.
Homozygous factor V Leiden mutation in type IV Ehlers-Danlos patient
Refaat, Marwan; Hotait, Mostafa; Winston, Brion
2014-01-01
Ehlers-Danlos syndrome (EDS) is a group of inherited connective tissue disorders caused by collagen synthesis defects. Several hemostatic abnormalities have been described in EDS patients that increase the bleeding tendencies of these patients. This case report illustrates a patient with an unusual presentation of a patient with type IV EDS, platelet δ-storage pool disease and factor V Leiden mutation. Young woman having previous bilateral deep vein thrombosis and pulmonary emboli coexisting with ruptured splenic aneurysm and multiple other aneurysms now presented with myocardial infarction. Presence of factor V Leiden mutation raises the possibility that the infarct was due to acute coronary thrombosis, although coronary artery aneurysm and dissection with myocardial infarction is known to occur in vascular type EDS. This is the first report in the medical literature of factor V Leiden mutation in an EDS patient which made the management of our patient challenging with propensity to both bleeding and clotting. PMID:24653990
Kao, Yu-Chun; Bailey, Andy; Samminger, Bernhard; Tanimoto, Junji; Burnouf, Thierry
2016-07-01
Pooled human platelet lysate (HPL) is becoming the new gold standard as supplement for ex vivo cell culture for clinical protocols. However, the risk of pathogen contamination of HPL increases with the platelet pool size. We hypothesized that hollow fiber anion exchange membrane chromatography using QyuSpeed D (QSD) could remove resistant and untested bloodborne pathogens, such as parvoviruses and prions, from HPL-supplemented growth media without substantially affecting their capacity to support ex vivo cell expansion. Frozen or thawed platelet concentrates were serum-converted and centrifuged for obtaining HPL that was added to various growth media (ca. 100 mL), filtered through a 0.6-mL QSD membrane and characterized for proteins, growth factors and chemical composition. Capacity to expand Chinese hamster ovary, periodontal ligament, gingival fibroblast cells and Wharton's jelly mesenchymal stromal cells was studied. Removal of porcine parvovirus (PPV) and of the 263K prion strain of hamster-adapted scrapie was studied by spiking experiments following international guidelines. QSD had minimal impact on HPL-supplemented medium composition in proteins, growth factors and chemical content, nor capacity to expand and differentiate cells. In addition, QSD could remove ≥5.58 log10 [TCID50/mL] and ≥3.72 log10 of PPV and the 263K prion, respectively. QSD hollow fiber chromatography can be used to improve the virus and prion safety of HPL-supplemented media to safely expand cells for clinical protocols. These data bring new perspectives for increasingly safer use of pooled HPL in cell therapy and regenerative medicine applications. Copyright © 2016 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.
Aghabeigi, N; Lindsey, N; Zamani, A; Shishaeyan, B
2012-01-01
Background: The autoimmune disease known as Idiopathic (immune thrombocytopenic purpura thrombocytopenic purpura (ITP) is clinically defined by a low numbers of platelets in the circulation blood. This study aimed to isolate autoantibodies made against the platelet glycoproteins using platelets from healthy volunteers, to determine their specificity and further elucidate their effects on platelet function. Methods: This study used a phage display system to recognize Fab anti-platelet antibodies. Anti-platelet After isolation, the anti-platelet Fab-expressing phage was characterized by ELISA and Western blotting. The Fab-bearing phage pool obtained from five rounds of panning was analysed in order to determine its anti-platelet reactivity. Of the phage colonies obtained, 100 colonies of different sizes were randomly selected for reaction with whole platelets, using M13 phage as a negative control. Results: Twelve colonies of them had strong reactions against the whole platelet preparation, but only four colonies showed substantial reactivity against the lysed platelet preparation (lysate). Three of the four colonies showed three bands representing proteins with different molecular weights. The fourth colony showed only a single band. The final experiment to characterise the protein isolated from the phage library was a DNA gel agarose test. Conclusion: Each colony showed a DNA band that corresponded with the molecular size marker for 5.4 kbase pairs, and this suggested the presence of heavy and light antibody chains in the phage. PMID:23113135
Federal Register 2010, 2011, 2012, 2013, 2014
2011-07-07
... Project Dominion is proposing to extend the perimeter of its Woodhull Natural Gas Storage Pool (Pool... Pool Boundary Project and Request for Comments on Environmental Issues The staff of the Federal Energy... the environmental impacts of the proposed Woodhull Storage Pool Boundary Project (Project) involving...
Weymann, Alexander; Ali-Hasan-Al-Saegh, Sadeq; Sabashnikov, Anton; Popov, Aron-Frederik; Mirhosseini, Seyed Jalil; Nombela-Franco, Luis; Testa, Luca; Lotfaliani, Mohammadreza; Zeriouh, Mohamed; Liu, Tong; Dehghan, Hamidreza; Yavuz, Senol; de Oliveira Sá, Michel Pompeu Barros; Baker, William L.; Jang, Jae-Sik; Gong, Mengqi; Benedetto, Umberto; Dohmen, Pascal M.; D’Ascenzo, Fabrizio; Deshmukh, Abhishek J.; Biondi-Zoccai, Giuseppe; Calkins, Hugh; Stone, Gregg W.
2017-01-01
Background This systematic review with meta-analysis aimed to determine the strength of evidence for evaluating the association of platelet cellular and functional characteristics including platelet count (PC), MPV, platelet distribution width (PDW), platelet factor 4, beta thromboglobulin (BTG), and p-selectin with the occurrence of atrial fibrillation (AF) and consequent stroke. Material/Methods We conducted a meta-analysis of observational studies evaluating platelet characteristics in patients with paroxysmal, persistent and permanent atrial fibrillations. A comprehensive subgroup analysis was performed to explore potential sources of heterogeneity. Results Literature search of all major databases retrieved 1,676 studies. After screening, a total of 73 studies were identified. Pooled analysis showed significant differences in PC (weighted mean difference (WMD)=−26.93 and p<0.001), MPV (WMD=0.61 and p<0.001), PDW (WMD=−0.22 and p=0.002), BTG (WMD=24.69 and p<0.001), PF4 (WMD=4.59 and p<0.001), and p-selectin (WMD=4.90 and p<0.001). Conclusions Platelets play a critical and precipitating role in the occurrence of AF. Whereas distribution width of platelets as well as factors of platelet activity was significantly greater in AF patients compared to SR patients, platelet count was significantly lower in AF patients. PMID:28302997
Extended Storage of Pathogen-Reduced Platelet Concentrates (PRECON)
2016-10-01
J. Slichter, MD CONTRACTING ORGANIZATION: Bloodworks Northwest Seattle, WA 98104 REPORT DATE : October 2016 TYPE OF REPORT: Annual Report...NOT RETURN YOUR FORM TO THE ABOVE ADDRESS. 1. REPORT DATE October 2016 2. REPORT TYPE Annual 3. DATES COVERED 26-SEP-15 to 25-SEP-16 4. TITLE AND...Platelet Concentrates (PRECON) dated 7/21/14 • Protocol - Cold Apheresis Platelets in Isoplate (CAPI) • Quad Chart 10 Date : 07-21-2014 Project Title
Handling and storage of human body fluids for analysis of extracellular vesicles
Yuana, Yuana; Böing, Anita N.; Grootemaat, Anita E.; van der Pol, Edwin; Hau, Chi M.; Cizmar, Petr; Buhr, Egbert; Sturk, Auguste; Nieuwland, Rienk
2015-01-01
Because procedures of handling and storage of body fluids affect numbers and composition of extracellular vesicles (EVs), standardization is important to ensure reliable and comparable measurements of EVs in a clinical environment. We aimed to develop standard protocols for handling and storage of human body fluids for EV analysis. Conditions such as centrifugation, single freeze–thaw cycle, effect of time delay between blood collection and plasma preparation and storage were investigated. Plasma is the most commonly studied body fluid in EV research. We mainly focused on EVs originating from platelets and erythrocytes and investigated the behaviour of these 2 types of EVs independently as well as in plasma samples of healthy subjects. EVs in urine and saliva were also studied for comparison. All samples were analysed simultaneously before and after freeze–thawing by resistive pulse sensing, nanoparticle tracking analysis, conventional flow cytometry (FCM) and transmission (scanning) electron microscopy. Our main finding is that the effect of centrifugation markedly depends on the cellular origin of EVs. Whereas erythrocyte EVs remain present as single EVs after centrifugation, platelet EVs form aggregates, which affect their measured concentration in plasma. Single erythrocyte and platelet EVs are present mainly in the range of 100–200 nm, far below the lower limit of what can be measured by conventional FCM. Furthermore, the effects of single freeze–thaw cycle, time delay between blood collection and plasma preparation up to 1 hour and storage up to 1 year are insignificant (p>0.05) on the measured concentration and diameter of EVs from erythrocyte and platelet concentrates and EVs in plasma, urine and saliva. In conclusion, in standard protocols for EV studies, centrifugation to isolate EVs from collected body fluids should be avoided. Freezing and storage of collected body fluids, albeit their insignificant effects, should be performed identically for comparative EV studies and to create reliable biorepositories. PMID:26563735
Handling and storage of human body fluids for analysis of extracellular vesicles.
Yuana, Yuana; Böing, Anita N; Grootemaat, Anita E; van der Pol, Edwin; Hau, Chi M; Cizmar, Petr; Buhr, Egbert; Sturk, Auguste; Nieuwland, Rienk
2015-01-01
Because procedures of handling and storage of body fluids affect numbers and composition of extracellular vesicles (EVs), standardization is important to ensure reliable and comparable measurements of EVs in a clinical environment. We aimed to develop standard protocols for handling and storage of human body fluids for EV analysis. Conditions such as centrifugation, single freeze-thaw cycle, effect of time delay between blood collection and plasma preparation and storage were investigated. Plasma is the most commonly studied body fluid in EV research. We mainly focused on EVs originating from platelets and erythrocytes and investigated the behaviour of these 2 types of EVs independently as well as in plasma samples of healthy subjects. EVs in urine and saliva were also studied for comparison. All samples were analysed simultaneously before and after freeze-thawing by resistive pulse sensing, nanoparticle tracking analysis, conventional flow cytometry (FCM) and transmission (scanning) electron microscopy. Our main finding is that the effect of centrifugation markedly depends on the cellular origin of EVs. Whereas erythrocyte EVs remain present as single EVs after centrifugation, platelet EVs form aggregates, which affect their measured concentration in plasma. Single erythrocyte and platelet EVs are present mainly in the range of 100-200 nm, far below the lower limit of what can be measured by conventional FCM. Furthermore, the effects of single freeze-thaw cycle, time delay between blood collection and plasma preparation up to 1 hour and storage up to 1 year are insignificant (p>0.05) on the measured concentration and diameter of EVs from erythrocyte and platelet concentrates and EVs in plasma, urine and saliva. In conclusion, in standard protocols for EV studies, centrifugation to isolate EVs from collected body fluids should be avoided. Freezing and storage of collected body fluids, albeit their insignificant effects, should be performed identically for comparative EV studies and to create reliable biorepositories.
Progress in bio-manufacture of platelets for transfusion.
Heazlewood, Shen Y; Nilsson, Susan K; Cartledge, Kellie; Be, Cheang Ly; Vinson, Andrew; Gel, Murat; Haylock, David N
2017-11-01
Blood transfusion services face an ever-increasing demand for donor platelets to meet clinical needs. Whilst strategies for increasing platelet storage life and improving the efficiency of donor platelet collection are important, in the longer term, platelets generated by bio-manufacturing processes will be required to meet demands. Production of sufficient numbers of in vitro-derived platelets for transfusion represents a significant bioengineering challenge. In this review, we highlight recent progress in this area of research and outline the main technical and biological obstacles that need to be met before this becomes feasible and economic. A critical consideration is assurance of the functional properties of these cells as compared to their fresh, donor collected, counterparts. We contend that platelet-like particles and in vitro-derived platelets that phenotypically resemble fresh platelets must deliver the same functions as these cells upon transfusion. We also note recent progress with immortalized megakaryocyte progenitor cell lines, molecular strategies for reducing expression of HLA Class I to generate universal donor platelets and the move to early clinical studies with in vitro-derived platelets.
Platelet-rich therapies for musculoskeletal soft tissue injuries.
Moraes, Vinícius Y; Lenza, Mário; Tamaoki, Marcel Jun; Faloppa, Flávio; Belloti, João Carlos
2013-12-23
Platelet-rich therapies are being used increasingly in the treatment of musculoskeletal soft tissue injuries such as ligament, muscle and tendon tears and tendinopathies. These therapies can be used as the principal treatment or as an augmentation procedure (application after surgical repair or reconstruction). Platelet-rich therapies are produced by centrifuging a quantity of the patient's own blood and extracting the active, platelet-rich, fraction. The platelet-rich fraction is applied to the injured tissue; for example, by injection. Platelets have the ability to produce several growth factors, so these therapies should enhance tissue healing. There is a need to assess whether this translates into clinical benefit. To assess the effects (benefits and harms) of platelet-rich therapies for treating musculoskeletal soft tissue injuries. We searched the Cochrane Bone, Joint and Muscle Trauma Group Specialised Register (25 March 2013), the Cochrane Central Register of Controlled Trials (CENTRAL 2013 Issue 2), MEDLINE (1946 to March 2013), EMBASE (1980 to 2013 Week 12) and LILACS (1982 to March 2012). We also searched trial registers (to Week 2 2013) and conference abstracts (2005 to March 2012). No language or publication restrictions were applied. We included randomised and quasi-randomised controlled trials that compared platelet-rich therapy with either placebo, autologous whole blood, dry needling or no platelet-rich therapy for people with acute or chronic musculoskeletal soft tissue injuries. Primary outcomes were functional status, pain and adverse effects. Two review authors independently extracted data and assessed each study's risk of bias. Disagreement was resolved by discussion or by arbitration by a third author. We contacted trial authors for clarification of methods or missing data. Treatment effects were assessed using risk ratios for dichotomous data and mean differences (MD) or standardised mean differences (SMD) for continuous data, together with 95% confidence intervals. Where appropriate, data were pooled using the fixed-effect model for RR and MD, and the random-effects model for SMD. The quality of the evidence for each outcome was assessed using GRADE criteria. We included data from 19 small single centre trials (17 randomised and two quasi-randomised; 1088 participants) that compared platelet-rich therapy with placebo, autologous whole blood, dry needling or no platelet-rich therapy. These trials covered eight clinical conditions: rotator cuff tears (arthroscopic repair) (six trials); shoulder impingement syndrome surgery (one trial); elbow epicondylitis (three trials); anterior cruciate ligament (ACL) reconstruction (four trials), ACL reconstruction (donor graft site application) (two trials), patellar tendinopathy (one trial), Achilles tendinopathy (one trial) and acute Achilles rupture surgical repair (one trial). We also grouped trials into 'tendinopathies' where platelet-rich therapy (PRT) injections were the main treatment (five trials), and surgical augmentation procedures where PRT was applied during surgery (14 trials). Trial participants were mainly male, except in trials including rotator cuff tears, and elbow and Achilles tendinopathies.Three trials were judged as being at low risk of bias; the other 16 were at high or unclear risk of bias relating to selection, detection, attrition or selective reporting, or combinations of these. The methods of preparing platelet-rich plasma (PRP) varied and lacked standardisation and quantification of the PRP applied to the patient.We were able to pool data for our primary outcomes (function, pain, adverse events) for a maximum of 11 trials and 45% of participants. The evidence for all primary outcomes was judged as being of very low quality.Data assessing function in the short term (up to three months) were pooled from five trials that assessed PRT in three clinical conditions and used four different measures. These showed no significant difference between PRT and control (SMD 0.24; 95% confidence interval (CI) -0.07 to 0.56; P value 0.13; I² = 35%; 273 participants; positive values favour PRT). Medium-term function data (at six months) were pooled from six trials that assessed PRT in five clinical conditions and used six different measures. These also showed no difference between groups (SMD 0.06; 95% CI -0.39 to 0.51; P value 0.79; I² = 64%; 262 participants). Long-term function data (at one year) were pooled from 10 trials that assessed PRT in five clinical conditions and used six different measures. These also showed no difference between groups (SMD 0.25, 95% CI -0.07 to 0.57; P value 0.12; I² = 66%; 484 participants). Although the 95% confidence intervals indicate the possibility of a slightly poorer outcome in the PRT group up to a moderate difference in favour of PRT at short- and long-term follow-up, these do not translate into clinically relevant differences.Data pooled from four trials that assessed PRT in three clinical conditions showed a small reduction in short-term pain in favour of PRT on a 10-point scale (MD -0.95, 95% CI -1.41 to -0.48; I² = 0%; 175 participants). The clinical significance of this result is marginal.Four trials reported adverse events; another seven trials reported an absence of adverse events. There was no difference between treatment groups in the numbers of participants with adverse effects (7/241 versus 5/245; RR 1.31, 95% CI 0.48 to 3.59; I² = 0%; 486 participants).In terms of individual conditions, we pooled heterogeneous data for long-term function from six trials of PRT application during rotator cuff tear surgery. This showed no statistically or clinically significant differences between the two groups (324 participants). Pooled data for short-term function for three elbow epicondylitis trials (179 participants) showed a statistically significant difference in favour of PRT, but the clinical significance of this finding is uncertain.The available evidence is insufficient to indicate whether the effects of PRT will differ importantly in individual clinical conditions. Overall, and for the individual clinical conditions, there is currently insufficient evidence to support the use of PRT for treating musculoskeletal soft tissue injuries. Researchers contemplating RCTs should consider the coverage of currently ongoing trials when assessing the need for future RCTs on specific conditions. There is need for standardisation of PRP preparation methods.
Platelet-rich therapies for musculoskeletal soft tissue injuries.
Moraes, Vinícius Y; Lenza, Mário; Tamaoki, Marcel Jun; Faloppa, Flávio; Belloti, João Carlos
2014-04-29
Platelet-rich therapies are being used increasingly in the treatment of musculoskeletal soft tissue injuries such as ligament, muscle and tendon tears and tendinopathies. These therapies can be used as the principal treatment or as an augmentation procedure (application after surgical repair or reconstruction). Platelet-rich therapies are produced by centrifuging a quantity of the patient's own blood and extracting the active, platelet-rich, fraction. The platelet-rich fraction is applied to the injured tissue; for example, by injection. Platelets have the ability to produce several growth factors, so these therapies should enhance tissue healing. There is a need to assess whether this translates into clinical benefit. To assess the effects (benefits and harms) of platelet-rich therapies for treating musculoskeletal soft tissue injuries. We searched the Cochrane Bone, Joint and Muscle Trauma Group Specialised Register (25 March 2013), the Cochrane Central Register of Controlled Trials (CENTRAL 2013 Issue 2), MEDLINE (1946 to March 2013), EMBASE (1980 to 2013 Week 12) and LILACS (1982 to March 2012). We also searched trial registers (to Week 2 2013) and conference abstracts (2005 to March 2012). No language or publication restrictions were applied. We included randomised and quasi-randomised controlled trials that compared platelet-rich therapy with either placebo, autologous whole blood, dry needling or no platelet-rich therapy for people with acute or chronic musculoskeletal soft tissue injuries. Primary outcomes were functional status, pain and adverse effects. Two review authors independently extracted data and assessed each study's risk of bias. Disagreement was resolved by discussion or by arbitration by a third author. We contacted trial authors for clarification of methods or missing data. Treatment effects were assessed using risk ratios for dichotomous data and mean differences (MD) or standardised mean differences (SMD) for continuous data, together with 95% confidence intervals. Where appropriate, data were pooled using the fixed-effect model for RR and MD, and the random-effects model for SMD. The quality of the evidence for each outcome was assessed using GRADE criteria. We included data from 19 small single centre trials (17 randomised and two quasi-randomised; 1088 participants) that compared platelet-rich therapy with placebo, autologous whole blood, dry needling or no platelet-rich therapy. These trials covered eight clinical conditions: rotator cuff tears (arthroscopic repair) (six trials); shoulder impingement syndrome surgery (one trial); elbow epicondylitis (three trials); anterior cruciate ligament (ACL) reconstruction (four trials), ACL reconstruction (donor graft site application) (two trials), patellar tendinopathy (one trial), Achilles tendinopathy (one trial) and acute Achilles rupture surgical repair (one trial). We also grouped trials into 'tendinopathies' where platelet-rich therapy (PRT) injections were the main treatment (five trials), and surgical augmentation procedures where PRT was applied during surgery (14 trials). Trial participants were mainly male, except in trials including rotator cuff tears, and elbow and Achilles tendinopathies.Three trials were judged as being at low risk of bias; the other 16 were at high or unclear risk of bias relating to selection, detection, attrition or selective reporting, or combinations of these. The methods of preparing platelet-rich plasma (PRP) varied and lacked standardisation and quantification of the PRP applied to the patient.We were able to pool data for our primary outcomes (function, pain, adverse events) for a maximum of 11 trials and 45% of participants. The evidence for all primary outcomes was judged as being of very low quality.Data assessing function in the short term (up to three months) were pooled from four trials that assessed PRT in three clinical conditions and used four different measures. These showed no significant difference between PRT and control (SMD 0.26; 95% confidence interval (CI) -0.19 to 0.71; P value 0.26; I² = 51%; 162 participants; positive values favour PRT). Medium-term function data (at six months) were pooled from five trials that assessed PRT in five clinical conditions and used five different measures. These also showed no difference between groups (SMD -0.09, 95% CI -0.56 to 0.39; P value 0.72; I² = 50%; 151 participants). Long-term function data (at one year) were pooled from 10 trials that assessed PRT in five clinical conditions and used six different measures. These also showed no difference between groups (SMD 0.25, 95% CI -0.07 to 0.57; P value 0.12; I² = 66%; 484 participants). Although the 95% confidence intervals indicate the possibility of a poorer outcome in the PRT group up to a moderate difference in favour of PRT at short- and long-term follow-up, these do not translate into clinically relevant differences.Data pooled from four trials that assessed PRT in three clinical conditions showed a small reduction in short-term pain in favour of PRT on a 10-point scale (MD -0.95, 95% CI -1.41 to -0.48; I² = 0%; 175 participants). The clinical significance of this result is marginal.Four trials reported adverse events; another seven trials reported an absence of adverse events. There was no difference between treatment groups in the numbers of participants with adverse effects (7/241 versus 5/245; RR 1.31, 95% CI 0.48 to 3.59; I² = 0%; 486 participants).In terms of individual conditions, we pooled heterogeneous data for long-term function from six trials of PRT application during rotator cuff tear surgery. This showed no statistically or clinically significant differences between the two groups (324 participants).The available evidence is insufficient to indicate whether the effects of PRT will differ importantly in individual clinical conditions. Overall, and for the individual clinical conditions, there is currently insufficient evidence to support the use of PRT for treating musculoskeletal soft tissue injuries. Researchers contemplating RCTs should consider the coverage of currently ongoing trials when assessing the need for future RCTs on specific conditions. There is need for standardisation of PRP preparation methods.
Mazutis, Linas; Wu, Stephen; Sylman, Joanna L.; Ehrlicher, Allen; Machlus, Kellie R.; Feng, Qiang; Lu, Shijiang; Lanza, Robert; Neeves, Keith B.; Weitz, David A.; Italiano, Joseph E.
2014-01-01
Platelet transfusions total >2.17 million apheresis-equivalent units per year in the United States and are derived entirely from human donors, despite clinically significant immunogenicity, associated risk of sepsis, and inventory shortages due to high demand and 5-day shelf life. To take advantage of known physiological drivers of thrombopoiesis, we have developed a microfluidic human platelet bioreactor that recapitulates bone marrow stiffness, extracellular matrix composition, micro-channel size, hemodynamic vascular shear stress, and endothelial cell contacts, and it supports high-resolution live-cell microscopy and quantification of platelet production. Physiological shear stresses triggered proplatelet initiation, reproduced ex vivo bone marrow proplatelet production, and generated functional platelets. Modeling human bone marrow composition and hemodynamics in vitro obviates risks associated with platelet procurement and storage to help meet growing transfusion needs. PMID:25606631
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.
Effect of platelet lysate on human cells involved in different phases of wound healing.
Barsotti, Maria Chiara; Chiara Barsotti, Maria; Losi, Paola; Briganti, Enrica; Sanguinetti, Elena; Magera, Angela; Al Kayal, Tamer; Feriani, Roberto; Di Stefano, Rossella; Soldani, Giorgio
2013-01-01
Platelets are rich in mediators able to positively affect cell activity in wound healing. Aim of this study was to characterize the effect of different concentrations of human pooled allogeneic platelet lysate on human cells involved in the different phases of wound healing (inflammatory phase, angiogenesis, extracellular matrix secretion and epithelialization). Platelet lysate effect was studied on endothelial cells, monocytes, fibroblasts and keratinocytes, in terms of viability and proliferation, migration, angiogenesis, tissue repair pathway activation (ERK1/2) and inflammatory response evaluation (NFκB). Results were compared both with basal medium and with a positive control containing serum and growth factors. Platelet lysate induced viability and proliferation at the highest concentrations tested (10% and 20% v/v). Whereas both platelet lysate concentrations increased cell migration, only 20% platelet lysate was able to significantly promote angiogenic activity (p<0.05 vs. control), comparably to the positive control. Both platelet lysate concentrations activated important inflammatory pathways such as ERK1/2 and NFκB with the same early kinetics, whereas the effect was different for later time-points. These data suggest the possibility of using allogeneic platelet lysate as both an alternative to growth factors commonly used for cell culture and as a tool for clinical regenerative application for wound healing.
Effect of Platelet Lysate on Human Cells Involved in Different Phases of Wound Healing
Briganti, Enrica; Sanguinetti, Elena; Magera, Angela; Al Kayal, Tamer; Feriani, Roberto; Di Stefano, Rossella; Soldani, Giorgio
2013-01-01
Background Platelets are rich in mediators able to positively affect cell activity in wound healing. Aim of this study was to characterize the effect of different concentrations of human pooled allogeneic platelet lysate on human cells involved in the different phases of wound healing (inflammatory phase, angiogenesis, extracellular matrix secretion and epithelialization). Methodology/Principal Findings Platelet lysate effect was studied on endothelial cells, monocytes, fibroblasts and keratinocytes, in terms of viability and proliferation, migration, angiogenesis, tissue repair pathway activation (ERK1/2) and inflammatory response evaluation (NFκB). Results were compared both with basal medium and with a positive control containing serum and growth factors. Platelet lysate induced viability and proliferation at the highest concentrations tested (10% and 20% v/v). Whereas both platelet lysate concentrations increased cell migration, only 20% platelet lysate was able to significantly promote angiogenic activity (p<0.05 vs. control), comparably to the positive control. Both platelet lysate concentrations activated important inflammatory pathways such as ERK1/2 and NFκB with the same early kinetics, whereas the effect was different for later time-points. Conclusion/Significance These data suggest the possibility of using allogeneic platelet lysate as both an alternative to growth factors commonly used for cell culture and as a tool for clinical regenerative application for wound healing. PMID:24386412
Pathogen-Reduced, Extended Platelet Storage in Platelet Additive Solution (PAS)
2016-10-01
Sherrill J. Slichter, MD CONTRACTING ORGANIZATION: Bloodworks Northwest Seattle, WA 98104 REPORT DATE : October 2016 TYPE OF REPORT: Annual...PLEASE DO NOT RETURN YOUR FORM TO THE ABOVE ADDRESS. 1. REPORT DATE October 2016 2. REPORT TYPE Annual 3. DATES COVERED 15-SEP-15 to 14-SEP-16 4...the key research outcomes to date . Brrr Study Our study of platelets stored as whole blood at 4°C demonstrated that end-over-end rotation is required
Extended Storage of Pathogen Reduced Platelet Concentrates (PRECON)
2015-10-01
J. Slichter, MD CONTRACTING ORGANIZATION: Bloodworks Northwest Seattle, WA 98104 REPORT DATE : October 2015 TYPE OF REPORT: Annual PREPARED FOR...YOUR FORM TO THE ABOVE ADDRESS. 1. REPORT DATE October 2015 2. REPORT TYPE Annual 3. DATES COVERED 26 Sep 2014 - 25 Sep 2015 4. TITLE AND SUBTITLE...accomplishments to date . Brrr Study Our study of platelets stored as whole blood at 4°C demonstrated that end-over-end rotation is required to reduce platelet
Improved circulating microparticle analysis in acid-citrate dextrose (ACD) anticoagulant tube.
György, Bence; Pálóczi, Krisztina; Kovács, Alexandra; Barabás, Eszter; Bekő, Gabriella; Várnai, Katalin; Pállinger, Éva; Szabó-Taylor, Katalin; Szabó, Tamás G; Kiss, Attila A; Falus, András; Buzás, Edit I
2014-02-01
Recently extracellular vesicles (exosomes, microparticles also referred to as microvesicles and apoptotic bodies) have attracted substantial interest as potential biomarkers and therapeutic vehicles. However, analysis of microparticles in biological fluids is confounded by many factors such as the activation of cells in the blood collection tube that leads to in vitro vesiculation. In this study we aimed at identifying an anticoagulant that prevents in vitro vesiculation in blood plasma samples. We compared the levels of platelet microparticles and non-platelet-derived microparticles in platelet-free plasma samples of healthy donors. Platelet-free plasma samples were isolated using different anticoagulant tubes, and were analyzed by flow cytometry and Zymuphen assay. The extent of in vitro vesiculation was compared in citrate and acid-citrate-dextrose (ACD) tubes. Agitation and storage of blood samples at 37 °C for 1 hour induced a strong release of both platelet microparticles and non-platelet-derived microparticles. Strikingly, in vitro vesiculation related to blood sample handling and storage was prevented in samples in ACD tubes. Importantly, microparticle levels elevated in vivo remained detectable in ACD tubes. We propose the general use of the ACD tube instead of other conventional anticoagulant tubes for the assessment of plasma microparticles since it gives a more realistic picture of the in vivo levels of circulating microparticles and does not interfere with downstream protein or RNA analyses. Copyright © 2013 Elsevier Ltd. All rights reserved.
Multiwavelength UV/visible spectroscopy for the quantitative investigation of platelet quality
NASA Astrophysics Data System (ADS)
Mattley, Yvette D.; Leparc, German F.; Potter, Robert L.; Garcia-Rubio, Luis H.
1998-04-01
The quality of platelets transfused is vital to the effectiveness of the transfusion. Freshly prepared, discoid platelets are the most effective treatment for preventing spontaneous hemorrhage or for stopping an abnormal bleeding event. Current methodology for the routine testing of platelet quality involves random pH testing of platelet rich plasma and visual inspection of platelet rich plasma for a swirling pattern indicative of the discoid shape of the cells. The drawback to these methods is that they do not provide a quantitative and objective assay for platelet functionality that can be used on each platelet unit prior to transfusion. As part of a larger project aimed at characterizing whole blood and blood components with multiwavelength UV/vis spectroscopy, isolated platelets and platelet in platelet rich plasma have been investigated. Models based on Mie theory have been developed which allow for the extraction of quantitative information on platelet size, number and quality from multi-wavelength UV/vis spectra. These models have been used to quantify changes in platelet rich plasma during storage. The overall goal of this work is to develop a simple, rapid quantitative assay for platelet quality that can be used prior to platelet transfusion to ensure the effectiveness of the treatment. As a result of this work, the optical properties for isolated platelets, platelet rich plasma and leukodepleted platelet rich plasma have been determined.
Quebec platelet disorder: features, pathogenesis and treatment.
Diamandis, Maria; Veljkovic, D Kika; Maurer-Spurej, Elisabeth; Rivard, Georges E; Hayward, Catherine P M
2008-03-01
Quebec platelet disorder (QPD) is a rare, autosomal-dominant, inherited bleeding disorder that is associated with unique abnormalities in fibrinolysis. Its hallmark features are delayed-onset bleeding following hemostatic challenges that responds to fibrinolytic inhibitor therapy and increased expression and storage of the fibrinolytic enzyme urokinase plasminogen activator in platelets, without increased plasma urokinase plasminogen activator or systemic fibrinolysis. The increased urokinase plasminogen activator in QPD platelets is only partially inhibited, and, as a result, there is intraplatelet generation of plasmin, and secondary degradation of many platelet alpha-granule proteins. During clot formation, the urokinase plasminogen activator released by QPD platelets leads to platelet-dependent increased fibrinolysis, and this is postulated to be a major contributor to QPD bleeding. The focus of the present review is to summarize the current state of knowledge on QPD, including the history of this disorder, its clinical and laboratory features, and recommended approaches for its diagnosis and treatment.
Ródenas, J; Abarca, A; Gallardo, S
2011-08-01
BWR control rods are activated by neutron reactions in the reactor. The dose produced by this activity can affect workers in the area surrounding the storage pool, where activated rods are stored. Monte Carlo (MC) models for neutron activation and dose assessment around the storage pool have been developed and validated. In this work, the MC models are applied to verify the expected reduction of dose when the irradiated control rod is hanged in an inverted position into the pool. 2010 Elsevier Ltd. All rights reserved.
Platelet lysate obtained via plateletpheresis performed in standing and awake equine donors.
Sumner, Scarlett M; Naskou, Maria C; Thoresen, Merrilee; Copland, Ian; Peroni, John F
2017-07-01
Platelet preparations containing growth factors, attachment factors, and enzymes are appealing to enhance healing of injured tissues and as an alternative to xenogenic serum in cell culture media. Plateletpheresis is commonly used to collect platelets in human medicine but has not been validated in horses. Plateletpheresis to collect platelet concentrate was performed on six female, mixed breed, chemically restrained horses using commercially available apheresis equipment. Before and immediately after plateletpheresis, we performed physical examinations and collected blood for chemistry and coagulation panels and then again at 8, 16, 24, and 48 hours after the procedure. To produce platelet lysate, the platelet concentrate underwent two freeze-thaw cycles followed by centrifugation and filtration processing. The platelet lysate was then analyzed for cellular debris, fibrinogen, and growth factors. The collected platelet concentration contained a mean platelet yield of 390 × 10 3 /μL. Donor platelet count decreased from a mean of 193 × 10 3 /μL to 138 × 10 3 /μL after plateletpheresis, but no individual was at risk for hemorrhage. Pooled platelet lysate had minimal cellular residue and contained growth factor concentrations at 6.1 ng/mL for transforming growth factor-β1, at 3.5 ng/mL for platelet-derived growth factor-BB, and at 13.8 ng/mL for vascular endothelial growth factor-A. Plateletpheresis using commercially available apheresis equipment is a feasible option for collecting platelet concentrate from equine donors. The lysate generated from the apheresis product contains growth factors and has potential to be used as a fetal bovine serum substitute for cell culture. © 2017 AABB.
Code of Federal Regulations, 2011 CFR
2011-04-01
... plasma used for resuspension of the platelets shall be determined by the maintenance of a pH of not less than 6.2 during the storage period. The pH shall be measured on a sample of platelets which has been... an adverse effect upon the safety, purity, potency, or efficacy of the product. At the time of...
Code of Federal Regulations, 2013 CFR
2013-04-01
... plasma used for resuspension of the platelets shall be determined by the maintenance of a pH of not less than 6.2 during the storage period. The pH shall be measured on a sample of platelets which has been... an adverse effect upon the safety, purity, potency, or efficacy of the product. At the time of...
Zaer, F; Metz, S; Scornik, J C
1997-01-15
The enzyme-linked immunosorbent assay (ELISA) using HLA class I molecules purified from pooled platelets has the potential to detect HLA antibodies with increased efficiency without sacrificing sensitivity or specificity. This test, which was originally developed in our institution, has been independently validated by recent studies and is now commercially available. We now present evidence of its usefulness as a routine HLA antibody screening test for renal transplant patients. A total of 515 patients were tested monthly by ELISA (13.9 tests/patient) and by antiglobulin-enhanced panel reactivity (6.3 tests/patient). In patients found to be unsensitized, the incidence of false-positive results was less for ELISA than for the panel studies. In patients who were highly sensitized, both tests performed equally well, whereas discordant results were registered mainly in cases of mild sensitization. Because 66% of our patients were not sensitized, the ELISA was effective in reducing the number of more involved tests aimed at characterizing the antibodies. These results provide a foundation to use the pooled platelet HLA ELISA on a routine basis for HLA antibody screening.
Pathogen reduction of blood components.
Solheim, Bjarte G
2008-08-01
Thanks to many blood safety interventions introduced in developed countries the risk of transfusion transmitted infections has become exceedingly small in these countries. However, emerging pathogens still represent a serious challenge, as demonstrated by West Nile virus in the US and more recently by Chikungunya virus in the Indian Ocean. In addition bacterial contamination, particularly in platelets, and protozoa transmitted by blood components still represent sizeable risks in developed countries. In developing countries the risk of all transfusion transmitted infections is still high due to insufficient funding and organisation of the health service. Pathogen reduction of pooled plasma products has virtually eliminated the risk of transfusion transmitted infections, without compromising the quality of the products significantly. Pathogen reduction of blood components has been much more challenging. Solvent detergent treatment which has been so successfully applied for plasma products dissolves cell membranes, and can, therefore, only be applied for plasma and not for cellular blood components. Targeting of nucleic acids has been another method for pathogen inactivation of plasma and the only approach possible for cellular blood products. As documented in more than 15 year's track record, solvent detergent treatment of pooled plasma can yield high quality plasma. The increased risk for contamination by unknown viruses due to pooling is out weighed by elimination of TRALI, significant reduction in allergic reactions and standardisation of the product. Recently, a promising method for solvent detergent treatment of single donor plasma units has been published. Methylene blue light treatment of single donor plasma units has a similar long track record as pooled solvent detergent treated plasma; but the method is less well documented and affects coagulation factor activity more. Psoralen light treated plasma has only recently been introduced (CE marked in Europe, but not licensed by the FDA), while the method of Riboflavin light treatment of plasma still is under development. In addition to pathogen reduction the methods, however, result in some reduction of coagulation factor activity. For platelets only Psoralen and Riboflavin light treatment have been implemented. Both are CE marked products in Europe but only approved for clinical trials in the USA. The methods affect platelet activity, but result in clinically acceptable platelets with only slightly reduced CCI and increased demand for platelet transfusions. Pathogen reduction of red blood cells with FRALE (S-303) or INACTINE (PEN110) has so far resulted in the formation of antibodies against neo-epitopes on red blood cells. A promising method for Riboflavin treatment of red blood cells is under development. This manuscript reviews the current experience and discusses future trends.
Ng, Ashley P.; Kauppi, Maria; Metcalf, Donald; Hyland, Craig D.; Josefsson, Emma C.; Lebois, Marion; Zhang, Jian-Guo; Baldwin, Tracey M.; Di Rago, Ladina; Hilton, Douglas J.; Alexander, Warren S.
2014-01-01
Thrombopoietin (TPO) acting via its receptor, the cellular homologue of the myeloproliferative leukemia virus oncogene (Mpl), is the major cytokine regulator of platelet number. To precisely define the role of specific hematopoietic cells in TPO-dependent hematopoiesis, we generated mice that express the Mpl receptor normally on stem/progenitor cells but lack expression on megakaryocytes and platelets (MplPF4cre/PF4cre). MplPF4cre/PF4cre mice displayed profound megakaryocytosis and thrombocytosis with a remarkable expansion of megakaryocyte-committed and multipotential progenitor cells, the latter displaying biological responses and a gene expression signature indicative of chronic TPO overstimulation as the underlying causative mechanism, despite a normal circulating TPO level. Thus, TPO signaling in megakaryocytes is dispensable for platelet production; its key role in control of platelet number is via generation and stimulation of the bipotential megakaryocyte precursors. Nevertheless, Mpl expression on megakaryocytes and platelets is essential to prevent megakaryocytosis and myeloproliferation by restricting the amount of TPO available to stimulate the production of megakaryocytes from the progenitor cell pool. PMID:24711413
Ng, Ashley P; Kauppi, Maria; Metcalf, Donald; Hyland, Craig D; Josefsson, Emma C; Lebois, Marion; Zhang, Jian-Guo; Baldwin, Tracey M; Di Rago, Ladina; Hilton, Douglas J; Alexander, Warren S
2014-04-22
Thrombopoietin (TPO) acting via its receptor, the cellular homologue of the myeloproliferative leukemia virus oncogene (Mpl), is the major cytokine regulator of platelet number. To precisely define the role of specific hematopoietic cells in TPO-dependent hematopoiesis, we generated mice that express the Mpl receptor normally on stem/progenitor cells but lack expression on megakaryocytes and platelets (Mpl(PF4cre/PF4cre)). Mpl(PF4cre/PF4cre) mice displayed profound megakaryocytosis and thrombocytosis with a remarkable expansion of megakaryocyte-committed and multipotential progenitor cells, the latter displaying biological responses and a gene expression signature indicative of chronic TPO overstimulation as the underlying causative mechanism, despite a normal circulating TPO level. Thus, TPO signaling in megakaryocytes is dispensable for platelet production; its key role in control of platelet number is via generation and stimulation of the bipotential megakaryocyte precursors. Nevertheless, Mpl expression on megakaryocytes and platelets is essential to prevent megakaryocytosis and myeloproliferation by restricting the amount of TPO available to stimulate the production of megakaryocytes from the progenitor cell pool.
The origin and function of platelet glycosyltransferases
Rumjantseva, Viktoria; Sørensen, Anne Louise Tølbøll; Patel-Hett, Sunita; Josefsson, Emma C.; Bennett, Eric P.; Italiano, Joseph E.; Clausen, Henrik; Hartwig, John H.; Hoffmeister, Karin M.
2012-01-01
Platelets are megakaryocyte subfragments that participate in hemostatic and host defense reactions and deliver pro- and antiangiogenic factors throughout the vascular system. Although they are anucleated cells that lack a complex secretory apparatus with distinct Golgi/endoplasmic reticulum compartments, past studies have shown that platelets have glycosyltransferase activities. In the present study, we show that members of 3 distinct glycosyltransferase families are found within and on the surface of platelets. Immunocytology and flow cytometry results indicated that megakaryocytes package these Golgi-derived glycosyltransferases into vesicles that are sent via proplatelets to nascent platelets, where they accumulate. These glycosyltransferases are active, and intact platelets glycosylate large exogenous substrates. Furthermore, we show that activation of platelets results in the release of soluble glycosyltransferase activities and that platelets contain sufficient levels of sugar nucleotides for detection of glycosylation of exogenously added substrates. Therefore, the results of the present study show that blood platelets are a rich source of both glycosyltransferases and donor sugar substrates that can be released to function in the extracellular space. This platelet-glycosylation machinery offers a pathway to a simple glycoengineering strategy improving storage of platelets and may serve hitherto unknown biologic functions. PMID:22613794
Guo, Ya-Huan; Sun, Hai-Feng; Zhang, Yan-Bing; Liao, Zi-Jun; Zhao, Lei; Cui, Jie; Wu, Tao; Lu, Jian-Rong; Nan, Ke-Jun; Wang, Shu-Hong
2017-03-21
Conflicting evidence exists regarding the effects of platelet/lymphocyte ratio (PLR) and lymphocyte/monocyte ratio(LMR) on the prognosis of colorectal cancer (CRC) patients. This study aimed to evaluate the roles of the PLR and LMR in predicting the prognosis of CRC patients via meta-analysis. Eligible studies were retrieved from the PubMed, Embase,andChina National Knowledge Infrastructure (CNKI) databases, supplemented by a manual search of references from retrieved articles. Pooled hazard ratios (HR) with 95% confidence intervals (95% CI) were calculated using the generic inverse variance and random-effect model to evaluate the association of PLR and LMR with prognostic variables in CRC, including overall survival (OS), cancer-specific survival (CSS) and disease-free survival (DFS). Thirty-three studies containing 15,404 patients met criteria for inclusion. Pooled analysis suggested that elevated PLR was associated with poorer OS (pooled HR = 1.57, 95% CI: 1.41 - 1.75, p< 0.00001, I2=26%) and DFS (pooled HR = 1.58, 95% CI: 1.31 - 1.92, p< 0.00001, I2=66%). Conversely, high LMR correlated with more favorable OS (pooled HR = 0.59, 95% CI: 0.50 - 0.68, p< 0.00001, I2=44%), CSS (pooled HR = 0.54, 95% CI: 0.40 - 0.72, p< 0.00001, I2=11%) and DFS (pooled HR = 0.82, 95% CI: 0.71- 0.94,p=0.005, I2=29%). Elevated PLR was associated with poor prognosis, while high LMR correlated with more favorable outcomes in CRC patients. Pretreatment PLR and LMR could serve as prognostic predictors in CRC patients.
Human platelet lysate: Replacing fetal bovine serum as a gold standard for human cell propagation?
Burnouf, Thierry; Strunk, Dirk; Koh, Mickey B C; Schallmoser, Katharina
2016-01-01
The essential physiological role of platelets in wound healing and tissue repair builds the rationale for the use of human platelet derivatives in regenerative medicine. Abundant growth factors and cytokines stored in platelet granules can be naturally released by thrombin activation and clotting or artificially by freeze/thaw-mediated platelet lysis, sonication or chemical treatment. Human platelet lysate prepared by the various release strategies has been established as a suitable alternative to fetal bovine serum as culture medium supplement, enabling efficient propagation of human cells under animal serum-free conditions for a multiplicity of applications in advanced somatic cell therapy and tissue engineering. The rapidly increasing number of studies using platelet derived products for inducing human cell proliferation and differentiation has also uncovered a considerable variability of human platelet lysate preparations which limits comparability of results. The main variations discussed herein encompass aspects of donor selection, preparation of the starting material, the possibility for pooling in plasma or additive solution, the implementation of pathogen inactivation and consideration of ABO blood groups, all of which can influence applicability. This review outlines the current knowledge about human platelet lysate as a powerful additive for human cell propagation and highlights its role as a prevailing supplement for human cell culture capable to replace animal serum in a growing spectrum of applications. Copyright © 2015 Elsevier Ltd. All rights reserved.
Influence of cardiopulmonary bypass on platelet and neutrophil accumulations in internal organs.
Bhujle, R; Li, J; Shastri, P; Gaffke, J N; Clift, J E; Ye, Y W; Dollar, M L; Ching, P; Chao, R; Constantinescu, A; Kulkarni, P V; Cheng, Q C; Wians, F; Jessen, M E; Eberhart, R C
1997-01-01
The authors employed gamma scintigraphy to quantify the post bypass accumulations of platelets and neutrophils in the lung, liver, and heart of adult pigs subjected to a standard 90 min regimen of normothermic cardiopulmonary bypass (CPB). Coated and uncoated microporous polypropylene oxygenator circuits were studied for Cobe Duo (Arvada, CO) oxygenators (amphophilic silicone-caprolactone oligomer [SMA] coating, n = 8 each) and Medtronic Maxima (Irvine, CA) oxygenators (Carmeda heparin coating, n = 5 each). Images of cells in the organs (deposited + blood pool) were corrected for tissue absorption and other factors and compared for a 2 hr period post CPB, using repeat measures ANOVA and rank tests. Platelet accumulations in internal organs correlated positively with whole blood platelet counts and negatively with platelet deposits in oxygenators during CPB. In general, uncoated CPB circuits significantly reduced platelet and neutrophil accumulations in lung, liver, and heart versus preCPB controls for the post CPB interval, for both systems. The SMA treatment significantly increased platelet accumulations versus uncoated controls in lung, liver, and heart for the 2 hr period, including the majority of the post CPB sampling intervals; platelet densities did not reach preCPB levels. Neutrophil accumulations were unaffected by the SMA coating. Carmeda heparin treatment significantly increased platelet accumulations in the liver, but not lung or heart. Despite preservation of circulating neutrophils observed with the Carmeda heparin treatment, neutrophil accumulations in internal organs were not elevated post CPB.
Russell, K A; Koch, T G
2016-03-01
Multipotent mesenchymal stromal cells (MSC) are often culture-expanded in vitro. Presently, expansion medium (EM) for MSC is supplemented with fetal bovine serum (FBS). However, increasing cost, variable composition and potential risks associated with bovine antigens call for alternatives. Platelet lysate (PL) has shown promise as an alternative supplement. To determine how equine umbilical cord blood (CB) MSC proliferate in EM enriched with PL or FBS at various concentrations. Randomised dose escalation study. Platelet concentrate was generated from 5 equine whole blood samples through a double centrifugation method and standardised to 1 × 10(12) platelets/l prior to a freeze/thaw cycle to produce PL. Pooled PL or pooled FBS was added to EM at concentrations of 5% to 60%. Proliferation of 4 equine CB-MSC cultures was determined after 4 days using a resazurin semiquantitative assay. Cord blood-MSC proliferated with a dose-dependent response with no significant difference found between PL and FBS up to a 30% concentration. Beyond 30%, proliferation fell in the PL-cultured cells, while continued dose-dependent proliferation was noted in the FBS-cultured cells. Despite reduced cell numbers in high PL concentrations, live/dead staining revealed that adherent cells remained viable. Expansion medium enriched with PL can support short-term equine CB-MSC proliferation at conventional culture concentrations. Based on the unexpected suppression of CB-MSC at higher PL concentrations, an in vivo dose study is indicated to investigate if combinational therapies of CB-MSC and platelet-rich plasma are associated with synergistic or antagonistic effect on CB-MSC function. © 2015 EVJ Ltd.
A&M. TAN607. Detail of fuel storage pool under construction. Camera ...
A&M. TAN-607. Detail of fuel storage pool under construction. Camera is on berm and facing northwest. Note depth of excavation. Formwork underway for floor and concrete walls of pool; wall between pool and vestibule. At center left of view, foundation for liquid waste treatment plant is poured. Date: August 25, 1953. INEEL negative no. 8541 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID
Propionibacterium acnes lacks the capability to proliferate in platelet concentrates.
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.
Wilson, Brooke H; Cole, Brian J; Goodale, Margaret B; Fortier, Lisa A
2018-04-01
The aim of this study was to provide clinical recommendations about the use of platelet-rich plasma (PRP) that was subjected to short-term storage at room temperature. We determined bioactive growth factor and cytokine concentrations as indicators of platelet and white blood cell degranulation in blood and PRP. Additionally, this study sought to validate the use of manual, direct smear analysis as an alternative to automated methods for platelet quantification in PRP. Blood was used to generate low-leukocyte PRP (Llo PRP) or high-leukocyte PRP (Lhi PRP). Blood was either processed immediately or kept at room temperature for 2 or 4 hours prior to generation of PRP, which was then held at room temperature for 0, 1, 2, or 4 hours. Subsequently, bioactive transforming growth factor beta-1 and matrix metalloproteinase-9 were measured by ELISA (enzyme-linked immunosorbent assay). Manual and automated platelet counts were performed on all blood and PRP samples. There were no differences in growth factor or cytokine concentration when blood or Llo PRP or Lhi PRP was retained at room temperature for up to 4 hours. Manual, direct smear analysis for platelet quantification was not different from the use of automated machine counting for PRP samples, but in the starting blood samples, manual platelet counts were significantly higher than those generated using automated technology. When there is a delay of up to 4 hours in the generation of PRP from blood or in the application of PRP to the patient, bioactive growth factor and cytokine concentrations remain stable in both blood and PRP. A manual direct counting method is a simple, cost-effective, and valid method to measure the contents of the PRP product being delivered to the patient.
Wang, Shichun; Jiang, Tianlun; Fan, Yahan; Zhao, Shuming
2018-03-29
Cryopreservation can slow down the metabolism and decrease the risk of bacterial contamination. But, chilled platelets (PLTs) show a reduced period in circulation due to the rapid clearance by hepatic cells or spleen macrophages after transfusion. The deleterious changes that PLTs undergo are mainly considered the result of PLT protein variation. However, the basis for proteomic variation of stored PLTs remains poorly understood. Besides count, activation markers (CD62P and Annexin V), and aggregation, we used quantitative mass spectrometry to create the first comprehensive and quantitative human PLT proteome of samples stored at different temperatures (22°C, 10°C and -80°C). We found different conditions caused different platelet storage lesion (PSL). PLT count was decreased no matter at what temperature stored. PLTs viability at low temperature dropped by 21.78% and 11.21%, respectively, as compared 10.26% at room temperature, there were no significant differences between the storage methods. Membrane expression of CD62P gradually increased in all groups especially stored at 22°C up to 40% and 10°C up to 30%. However, exposure of PS on the PLT membrane was below 1% in every group. The PLT proteome showed there were 575 and 454 potential proteins identified by general iTRAQ analysis and phosphorylation iTRAQ a nalysis, respectively, among them, 33 common differentially expressed proteins caused by storage time and 44 caused by storage temperature Especially, membrane-bound proteins (such as FERMT3, STX4, MYL9 and TAGLN2) played key roles in PLT storage lesion. The pathways "Endocytosis", "Fc gamma R-mediated phagocytosis" and "Regulation of actin cytoskeleton" were affected predominantly by storage time. And the pathways "SNARE interactions in vesicular transport" and "Vasopressin-regulated water reabsorption" were affected by cold storage in our study. Proteomic results can help us to understand PLT biochemistry and physiology and thus unravel the mechanisms of PSL in time and space for more successful PLT transfusion therapy.
Gao, Xiao-Ming; Moore, Xiao-Lei; Liu, Yang; Wang, Xin-Yu; Han, Li-Ping; Su, Yidan; Tsai, Alan; Xu, Qi; Zhang, Ming; Lambert, Gavin W; Kiriazis, Helen; Gao, Wei; Dart, Anthony M; Du, Xiao-Jun
2016-07-01
Acute myocardial infarction (AMI) is characterized by a rapid increase in circulating platelet size but the mechanism for this is unclear. Large platelets are hyperactive and associated with adverse clinical outcomes. We determined mean platelet volume (MPV) and platelet-monocyte conjugation (PMC) using blood samples from patients, and blood and the spleen from mice with AMI. We further measured changes in platelet size, PMC, cardiac and splenic contents of platelets and leucocyte infiltration into the mouse heart. In AMI patients, circulating MPV and PMC increased at 1-3 h post-MI and MPV returned to reference levels within 24 h after admission. In mice with MI, increases in platelet size and PMC became evident within 12 h and were sustained up to 72 h. Splenic platelets are bigger than circulating platelets in normal or infarct mice. At 24 h post-MI, splenic platelet storage was halved whereas cardiac platelets increased by 4-fold. Splenectomy attenuated all changes observed in the blood, reduced leucocyte and platelet accumulation in the infarct myocardium, limited infarct size and alleviated cardiac dilatation and dysfunction. AMI-induced elevated circulating levels of adenosine diphosphate and catecholamines in both human and the mouse, which may trigger splenic platelet release. Pharmacological inhibition of angiotensin-converting enzyme, β1-adrenergic receptor or platelet P2Y12 receptor reduced platelet abundance in the murine infarct myocardium albeit having diverse effects on platelet size and PMC. In conclusion, AMI evokes release of splenic platelets, which contributes to the increase in platelet size and PMC and facilitates myocardial accumulation of platelets and leucocytes, thereby promoting post-infarct inflammation. © 2016 The Author(s). published by Portland Press Limited on behalf of the Biochemical Society.
Lu, Shi-Jiang; Li, Feng; Yin, Hong; Feng, Qiang; Kimbrel, Erin A; Hahm, Eunsil; Thon, Jonathan N; Wang, Wei; Italiano, Joseph E; Cho, Jaehyung; Lanza, Robert
2011-01-01
Platelets play an essential role in hemostasis and atherothrombosis. Owing to their short storage time, there is constant demand for this life-saving blood component. In this study, we report that it is feasible to generate functional megakaryocytes and platelets from human embryonic stem cells (hESCs) on a large scale. Differential-interference contrast and electron microscopy analyses showed that ultrastructural and morphological features of hESC-derived platelets were indistinguishable from those of normal blood platelets. In functional assays, hESC-derived platelets responded to thrombin stimulation, formed microaggregates, and facilitated clot formation/retraction in vitro. Live cell microscopy demonstrated that hESC-platelets formed lamellipodia and filopodia in response to thrombin activation, and tethered to each other as observed in normal blood. Using real-time intravital imaging with high-speed video microscopy, we have also shown that hESC-derived platelets contribute to developing thrombi at sites of laser-induced vascular injury in mice, providing the first evidence for in vivo functionality of hESC-derived platelets. These results represent an important step toward generating an unlimited supply of platelets for transfusion. Since platelets contain no genetic material, they are ideal candidates for early clinical translation involving human pluripotent stem cells. PMID:21221130
Thibaut, Julien; Mérieux, Yves; Rigal, Dominique; Gillet, Germain
2012-01-01
Background Neonatal alloimmune thrombocytopenia is mostly due to the presence of maternal antibodies against the fetal platelet antigen HPA-1a on the platelet integrin GPIIb-IIIa. Accurate detection of anti-HPA-1a antibodies in the mother is, therefore, critical. Current diagnostic assays rely on the availability of pools of human platelets that vary according to donors and blood centers. There is still no satisfactory standardization of these assays. Design and Methods Peptide aptamer was used to detect and identify HPA-1a-specific antibodies in human serum that do not require human platelets. A peptide aptamer library was screened using an anti-HPA-1a human monoclonal antibody as a bait to isolate an aptamer that mimics the human platelet antigen HPA-1a. Results This is the first report in platelet immunology of the use of a peptide aptamer for diagnostic purposes. This assay gives better results than the MAIPA currently in use, detecting around 90% of the expected alloantibodies. Conclusions This assay could help define a standard for the quantitation of anti-HPA antibodies. This report also demonstrates that peptide aptamers can potentially detect a variety of biomarkers in body fluids; this is of particular interest for diagnostic purposes. PMID:22133781
Sebastian, Roby; Ratliff, Todd; Winch, Peter D; Tumin, Dmitry; Gomez, Daniel; Tobias, Joseph; Galantowicz, Mark; Naguib, Aymen N
2017-01-01
The majority of allogeneic transfusions occur in the perioperative setting, especially during cardiac surgery. In addition to the economic implications, there is emerging evidence that blood transfusion may increase both morbidity and mortality. Acute normovolemic hemodilution (ANH) may limit the need for blood products. The primary objective of this study was to determine if the method of blood collection (syringe or bag) during the ANH process impacted the platelet count and function. The secondary objectives included the need for perioperative blood transfusions during the procedure and in the intensive care unit. In addition, we assessed these outcomes' associations with ANH parameters including the method of collection, time of storage, and volume removed. Data were collected prospectively from 50 patients undergoing cardiac surgery on cardiopulmonary bypass over a 6-month period. Platelet count and function were measured for the ANH blood immediately after collection and again prior to transfusing to the patient at the end of cardiopulmonary bypass. Other data collected included ANH volume, length of storage, and the quantity of all blood products given throughout the perioperative period. No change in platelet count or function was noted regardless of the length of time or collection method for the ANH blood. Twenty-three patients received blood or blood products in the operating room or the intensive care unit, while 27 patients received no blood transfusion during their entire hospitalization. Higher ANH volume (ml·kg -1 ) and longer storage time were associated with a greater need for intraoperative transfusions. Acute normovolemic hemodilution protects the platelets from the untoward effects of cardiopulmonary bypass and offers an important autologous blood product that improves hemostasis at the conclusion of surgery. Platelet count and function are preserved regardless of the method of collection or the length of storage. The volume of ANH removed appears to be an important determinant of blood product use and further understanding of the impact of this variable is a future direction of upcoming prospective research. © 2016 John Wiley & Sons Ltd.
Davey, Sue; Navarrete, Cristina; Brown, Colin
2017-06-01
Twenty-nine human platelet antigen systems have been described to date, but the majority of current genotyping methods are restricted to the identification of those most commonly associated with alloantibody production in a clinical context. This can result in a protracted investigation if causative human platelet antigens are rare or novel. A targeted next-generation sequencing approach was designed to detect all known human platelet antigens with the additional capability of identifying novel mutations in the encoding genes. A targeted enrichment, high-sensitivity HaloPlex assay was designed to sequence all exons and flanking regions of the six genes known to encode human platelet antigens. Indexed DNA libraries were prepared from 47 previously human platelet antigen-genotyped samples and subsequently combined into one of three pools for sequencing on an Illumina MiSeq platform. The generated FASTQ files were aligned and scrutinized for each human platelet antigen polymorphism using SureCall data analysis software. Forty-six samples were successfully genotyped for human platelet antigens 1 through 29bw, with an average per base coverage depth of 1144. Concordance with historical human platelet antigen genotypes was 100%. A putative novel mutation in Exon 10 of the integrin β-3 (ITGB3) gene from an unsolved case of fetal neonatal alloimmune thrombocytopenia was also detected. A next-generation sequencing-based method that can accurately define all known human platelet antigen polymorphisms was developed. With the ability to sequence up to 96 samples simultaneously, our HaloPlex design could be used for high-throughput human platelet antigen genotyping. This method is also applicable for investigating fetal neonatal alloimmune thrombocytopenia when rare or novel human platelet antigens are suspected. © 2017 AABB.
Serra-Millàs, Montserrat
2016-01-01
Brain-derived neurotrophic factor (BDNF) plays an important role in central nervous system development, neurogenesis and neuronal plasticity. BDNF is also expressed in several non-neuronal tissues, and it could play an important role in other processes, such as cancer, angiogenesis, etc. Platelets are the major source of peripheral BDNF. However, platelets also contain high amounts of serotonin; they express specific surface receptors during activation, and a multitude of pro-inflammatory and immunomodulatory bioactive compounds are secreted from the granules. Until recently, there was insufficient knowledge regarding the relationship between BDNF and platelets. Recent studies showed that BDNF is present in two distinct pools in platelets, in α-granules and in the cytoplasm, and only the BDNF in the granules is secreted following stimulation, representing 30% of the total BDNF in platelets. BDNF has an important role in the pathophysiology of depression. Low levels of serum BDNF have been described in patients with major depressive disorder, and BDNF levels increased with chronic antidepressant treatment. Interestingly, there is an association between depression and platelet function. This review analyzed studies that evaluated the relationship between BDNF and platelet activation and the effect of treatments on both parameters. Only a few studies consider this possible confounding factor, and it could be very important in diseases such as depression, which show changes in both parameters. PMID:27014600
The Need for Integrating the Back End of the Nuclear Fuel Cycle in the United States of America
Bonano, Evaristo J.; Kalinina, Elena A.; Swift, Peter N.
2018-02-26
Current practice for commercial spent nuclear fuel management in the United States of America (US) includes storage of spent fuel in both pools and dry storage cask systems at nuclear power plants. Most storage pools are filled to their operational capacity, and management of the approximately 2,200 metric tons of spent fuel newly discharged each year requires transferring older and cooler fuel from pools into dry storage. In the absence of a repository that can accept spent fuel for permanent disposal, projections indicate that the US will have approximately 134,000 metric tons of spent fuel in dry storage by mid-centurymore » when the last plants in the current reactor fleet are decommissioned. Current designs for storage systems rely on large dual-purpose (storage and transportation) canisters that are not optimized for disposal. Various options exist in the US for improving integration of management practices across the entire back end of the nuclear fuel cycle.« less
The Need for Integrating the Back End of the Nuclear Fuel Cycle in the United States of America
DOE Office of Scientific and Technical Information (OSTI.GOV)
Bonano, Evaristo J.; Kalinina, Elena A.; Swift, Peter N.
Current practice for commercial spent nuclear fuel management in the United States of America (US) includes storage of spent fuel in both pools and dry storage cask systems at nuclear power plants. Most storage pools are filled to their operational capacity, and management of the approximately 2,200 metric tons of spent fuel newly discharged each year requires transferring older and cooler fuel from pools into dry storage. In the absence of a repository that can accept spent fuel for permanent disposal, projections indicate that the US will have approximately 134,000 metric tons of spent fuel in dry storage by mid-centurymore » when the last plants in the current reactor fleet are decommissioned. Current designs for storage systems rely on large dual-purpose (storage and transportation) canisters that are not optimized for disposal. Various options exist in the US for improving integration of management practices across the entire back end of the nuclear fuel cycle.« less
Balint, Bela; Vucetić, Dusan; Trajković-Lakić, Zlatija; Petakov, Marijana; Bugarski, Diana; Brajusković, Goran; Taseski, Jovan
2002-01-01
Cryopreservation of platelets is of great interest, since it could extend the shelf life of therapeutic platelet concentrates and facilitate stockpiling and inventory control in blood banking. Despite the use of many cryopreservation procedures the optimal cryopreservation procedure is not defined yet. We have compared the cryopreservation of human platelets by various protocols employing controlled-rate and non-controlled-rate freezing procedures in combination with different concentrations of DMSO (6% and 10%) or 5% DMSO + 6% HES combination. After storage for 1 to 3 months, samples were thawed and analyzed. Measurements included cell recovery, platelet viability according to hypotonic shock response (HSR), platelet aggregation with ADP, morphological and ultrastructural properties of defrozen platelets. Our findings show that the application of our original procedure for controlled-rate freezing consisting of six cooling steps (cooling rate 1 degree C/min) with compensation of released heat of fusion (cooling rate 2 degrees C/min) has significantly influenced the quality of thawed platelets. At the same time, a concentration of 6% DMSO proved to be the most effective. In summary, cryopreservation of human platelets using controlled-rate freezing procedure in combination with lower (6%) DMSO concentration resulted in less damage from freezing and higher recovered function of platelets.
The influence of platelets, plasma and red blood cells on functional haemostatic assays.
Bochsen, Louise; Johansson, Pär I; Kristensen, Annemarie T; Daugaard, Gedske; Ostrowski, Sisse R
2011-04-01
Functional whole blood haemostatic assays are used increasingly to guide transfusion therapy and monitor medical treatment and are also applied for in-vitro evaluations of the haemostatic potential of stored platelets. We investigated how the cellular and plasmatic elements, both isolated and combined, influenced the two methodologically different assays, thrombelastography (TEG) and impedance aggregometry (Multiplate). Platelet-rich plasma (200 × 10/l) or pure plasma (0 platelets), with and without added red blood cells (RBCs), hematocrit 0, 0.15 or 0.29, were produced in vitro from platelet concentrates, fresh frozen plasma and stored RBC. Pure platelets were investigated by removing plasma components from platelet concentrates by diafiltration against the platelet storage solution Intersol. Plasma was readded by diafiltration against plasma in Intersol. Haemostatic function was evaluated by TEG and Multiplate. In the TEG, increasing amounts of RBC reduced clot strength and clot kinetics (α-angle), most markedly in plasma/RBC without platelets. In contrast, RBC in a platelet concentrate matrix enhanced Multiplate aggregation in response to weak agonists (ADP and arachidonic acid). Furthermore, removing plasma from platelet concentrates eliminated the TEG response and diminished the Multiplate aggregation response, but readding plasma to the pure platelet concentrates restored the response. Each of the elements in whole blood, plasma, platelets and RBC, affected the Multiplate and TEG results differently. The results emphasize that the concentrations of all cellular and plasmatic components in whole blood should be taken into account when interpreting results obtained by TEG and multiplate.
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 consistent impact on cytokine content after PRP processing.
Detection of microbial contamination in platelets
NASA Astrophysics Data System (ADS)
Berg, Tracy L.; Leparc, German; Huffman, Debra E.; Gennaccaro, Angela L.; Garcia-Lopez, Alicia; Klungness, Greta; Stephans, Christie; Garcia-Rubio, Luis H.
2005-03-01
In the United States, approximately 100 patients develop fatal sepsis associated with platelet transfusions every year. Current culture methods take 24-48 hours to acquire results, which in turn decrease the shelf life of platelets. Many of the microorganisms that contaminate platelets can replicate easily at room temperature, which is the necessary storage temperature to keep platelets functional. Therefore, there is a need for in-situ quality control assessment of the platelet quality. For this purpose, a real time spectrophotometric technique has been developed. The Spectral Acquisition Processing Detection (SAPD) method, comprised of a UV-vis spectrophotometer and modeling algorithms, is a rapid method that can be performed prior to platelet transfusion to decrease the risk of bacterial infection to patients. The SAPD method has been used to determine changes in cell suspensions, based on size, shape, chemical composition and internal structure. Changes in these cell characteristics can in turn be used to determine microbial contamination, platelet aging and other physiologic changes. Detection limits of this method for platelet suspensions seeded with bacterial contaminants were identified to be less than 100 cfu/ml of sample. Bacterial counts below 1000 cfu/ml are not considered clinically significant. The SAPD method can provide real-time identification of bacterial contamination of platelets affording patients an increased level of safety without causing undue strain on laboratory budgets or personnel while increasing the time frame that platelets can be used by dramatically shortening contaminant detection time.
FlnA-null megakaryocytes prematurely release large and fragile platelets that circulate poorly
Jurak Begonja, Antonija; Hoffmeister, Karin M.; Hartwig, John H.
2011-01-01
Filamin A (FlnA) is a large cytoplasmic protein that crosslinks actin filaments and anchors membrane receptors and signaling intermediates. FlnAloxP PF4-Cre mice that lack FlnA in the megakaryocyte (MK) lineage have a severe macrothrombocytopenia because of accelerated platelet clearance. Macrophage ablation by injection of clodronate-encapsulated liposomes increases blood platelet counts in FlnAloxP PF4-Cre mice and reveals the desintegration of FlnA-null platelets into microvesicles, a process that occurs spontaneously during storage. FlnAloxP PF4-Cre bone marrows and spleens have a 2.5- to 5-fold increase in MK numbers, indicating increased thrombopoiesis in vivo. Analysis of platelet production in vitro reveals that FlnA-null MKs prematurely convert their cytoplasm into large CD61+ platelet-sized particles, reminiscent of the large platelets observed in vivo. FlnA stabilizes the platelet von Willebrand factor receptor, as surface expression of von Willebrand factor receptor components is normal on FlnA-null MKs but decreased on FlnA-null platelets. Further, FlnA-null platelets contain multiple GPIbα degradation products and have increased expression of the ADAM17 and MMP9 metalloproteinases. Together, the findings indicate that FlnA-null MKs prematurely release large and fragile platelets that are removed rapidly from the circulation by macrophages. PMID:21652675
Maurer-Spurej, Elisabeth; Larsen, Rune; Labrie, Audrey; Heaton, Andrew; Chipperfield, Kate
2016-08-01
In circulation, shedding of microparticles from a variety of viable cells can be triggered by pathological activation of inflammatory processes, by activation of coagulation or complement systems, or by physical stress. Elevated microparticle content (MPC) in donor blood might therefore indicate a clinical condition of the donor which, upon transfusion, might affect the recipient. In blood products, elevated MPC might also represent product stress. Surprisingly, the MPC in blood collected from normal blood donors is highly variable, which raises the question whether donor microparticles are present in-vivo and transfer into the final blood component, and how production methods and post-production processing might affect the MPC. We measured MPC using ThromboLUX in (a) platelet-rich plasma (PRP) of 54 apheresis donors and the corresponding apheresis products, (b) 651 apheresis and 646 pooled platelet concentrates (PCs) with plasma and 414 apheresis PCs in platelet additive solution (PAS), and (c) apheresis PCs before and after transportation, gamma irradiation, and pathogen inactivation (N = 8, 7, and 12 respectively). ThromboLUX-measured MPC in donor PRP and their corresponding apheresis PC samples were highly correlated (r = 0.82, P = .001). The average MPC in pooled PC was slightly lower than that in apheresis PC and substantially lower in apheresis PC stored with PAS rather than plasma. Mirasol Pathogen Reduction treatment significantly increased MPC with age. Thus, MPC measured in donor samples might be a useful predictor of product stability, especially if post-production processes are necessary. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.
Pidcoke, Heather F.; McFaul, Steve J.; Ramasubramanian, Anand K.; Parida, Bijaya K.; Mora, Alex G.; Fedyk, Chriselda G.; Valdez-Delgado, Krystal K.; Montgomery, Robbie K.; Reddoch, Kristin M.; Rodriguez, Armando C.; Aden, James K.; Jones, John A.; Bryant, Ron S.; Scherer, Michael R.; Reddy, Heather L.; Goodrich, Raymond P.; Cap, Andrew P.
2014-01-01
BACKGROUND Whole blood (WB) has been used in combat since World War I as it is readily available and replaces every element of shed blood. Component therapy has become standard; however, recent military successes with WB resuscitation have revived the debate regarding wider WB use. Characterization of optimal WB storage is needed. We hypothesized that refrigeration preserves WB function and that a pathogen reduction technology (PRT) based on riboflavin and ultraviolet light has no deleterious effect over 21 days of storage. STUDY DESIGN AND METHODS WB units were stored for 21 days either at 4°C or 22°C. Half of each temperature group underwent PRT, yielding four final treatment groups (n = 8 each): CON 4 (WB at 4°C); CON 22 (WB at 22°C); PRT 4 (PRT WB at 4°C); and PRT 22 (PRT WB at 22°C). Testing was at baseline, Days 1–7, 10, 14, and 21. Assays included coagulation factors; platelet activation, aggregation, and adhesion; and thromboelastography (TEG). RESULTS Prothrombin time (PT) and partial thromboplastin time increased over time; refrigeration attenuated the effects on PT (p ≤ 0.009). Aggregation decreased over time (p ≤ 0.001); losses were attenuated by refrigeration (p ≤ 0.001). Refrigeration preserved TEG parameters (p ≤ 0.001) and PRT 4 samples remained within normal limits throughout the study. Refrigeration in combination with PRT inhibited fibrinolysis (p ≤ 0.001) and microparticle formation (p ≤ 0.031). Cold storage increased shear-induced platelet aggregation and ristocetin-induced platelet agglutination (p ≥ 0.032), as well as GPIb-expressing platelets (p ≤ 0.009). CONCLUSION The in vitro hemostatic function of WB is largely unaffected by PRT treatment and better preserved by cold storage over 21 days. Refrigerated PRT WB may be suitable for trauma resuscitation. Clinical studies are warranted. PMID:23301966
Daurat, Aurélien; Roger, Claire; Gris, JeanChristophe; Daurat, Gérald; Feissel, Michel; Le Manach, Yannick; Lefrant, JeanYves; Muller, Laurent
2016-06-01
Controversy exists regarding the safety of the different types of platelet (PLT) concentrates. This study was aimed at comparing the rate of adverse reactions associated with apheresis PLT concentrates (APCs) and pooled PLT concentrates (PPCs) both in donors and in recipients. From the French national hemovigilance system, types and numbers of recipient adverse reactions were compared over a period from 2009 to 2011. Donor adverse reactions were available for 2010 and 2011. This study involved 23 of 26 French regions. Main outcomes were the rates of adverse reaction in recipients and serious adverse reaction in donors. There were 790,854 PLT transfusions during the study period (477,747 [60%] with APCs, 313,107 [40%] with PPCs). APCs were associated with more adverse reactions (6244 vs. 2469 per 1,000,000, p < 0.001) and more severe and life-threatening reactions (respectively, 241 vs. 131 per 1,000,000, p < 0.001; and 182 vs. 121 per 1,000,000, p = 0.04). Mortality rates due to an adverse transfusion reaction were similar (15 vs. 6 per 1,000,000, p = 0.5). In donors, the number of whole blood (WB) donations was 4,722,685 whereas 266,095 apheresis procedures were performed. Serious adverse reactions were more frequent for apheresis procedures than for WB donations (5445 vs. 803 per 1,000,000, p < 0.001). Our findings suggest that apheresis PLTs may be more hazardous than pooled PLTs both in recipients and in donors. This study calls for randomized trials to confirm or refute these results. © 2016 AABB.
Zhao, Zhe; Zhao, Xinrui; Lu, Jingjing; Xue, Jing; Liu, Peishu; Mao, Hongluan
2018-04-01
The systemic inflammatory response markers have been reported to be associated with the prognosis of various cancers. We conducted this meta-analysis of retrospective studies to evaluate and identify the prognostic impact of neutrophil to lymphocyte ratio (NLR) and platelet to lymphocyte ratio (PLR) on ovarian cancer. PubMed, EMBASE, and China National Knowledge Infrastructure databases were included to search for eligible studies. The following terms were used: "neutrophil to lymphocyte ratio", "NLR", "platelet to lymphocyte ratio", "PLR", "ovarian cancer", "ovary cancer", "ovarian carcinoma", "ovary carcinoma", "ovarian neoplasm", "ovary neoplasm", "ovarian tumor", and "ovary tumor". The random-effects model was chosen to estimate the pooled HR with 95% CI. Heterogeneity between studies was assessed by Higgins I 2 value. The stability and heterogeneity of studies were analyzed by sensitivity analysis. Publication bias was examined by Egger's test and Begg's test with the funnel plots. 13 studies consisting of 3467 patients were considered for meta-analysis. We found that the high NLR had a poor prognostic impact on OS and PFS in ovarian cancer, with a pooled HR 1.70, 95% CI 1.35-2.15 and HR 1.77, 95% CI 1.48-2.12, respectively. Similarly, the results showed the high PLR adversely affected OS and PFS in ovarian cancer, with a pooled HR 2.05, 95% CI 1.70-2.48 and HR 1.85, 95% CI 1.53-2.25, respectively. In conclusion, we found that both NLR and PLR had an unfavorable impact on PFS and OS of patients with ovarian cancer. Our meta-analysis supported that NLR/PLR could be effective prognostic predictors of ovarian cancer.
Blood-Banking Techniques for Plateletpheresis in Swine
Sondeen, Jill L; Prince, Malcolm D; Polykratis, Irene A; Hernandez, Orlando; Torres-Mendoza, Jaime; Guzman, Rodolfo De; Aden, James K; Dubick, Michael A
2014-01-01
During the past several years, trauma resuscitation in human patients has evolved from decreased use of crystalloids to increased use of blood products. Of high interest is the role of platelets in trauma resuscitation. Because conducting prehospital resuscitation in human trauma patients is very difficult, swine are often the animal model of choice for such studies because their coagulation and hemodynamic systems are similar to those in humans. However, consistent production of sufficient swine platelets for such studies has not previously been achieved. We developed a method for producing swine platelets by using standard human techniques and equipment. We assessed pH, pO2, pCO2, lactate, thromboelastography, and platelet aggregation over 5 d of storage to determine whether the swine platelet product met the American Association of Blood Banks (AABB) standards for transfusion. Swine platelets met AABB standards at 24 h but not at later time points. In addition, we fluorescently labeled nonautologous platelets and then measured their percentage recovery over 5 h (the time used in subsequent experimental studies) when transfused into a recipient pig. We showed that 80% of the platelets stored for 24 h remained in the circulation and increased the recipient pigs’ thromboelastographic responses, indicating that the platelets were viable and active. Therefore, swine platelets stored for 24 h by using standard human products met the AABB criteria and were functional. PMID:24827574
Acute effects of 30 minutes of exposure to a smartphone call on in vitro platelet function
Lippi, Giuseppe; Danese, Elisa; Brocco, Giorgio; Gelati, Matteo; Salvagno, Gian Luca; Montagnana, Martina
2017-01-01
Background Significant concerns are now regularly raised about the safety of excessive mobile phone use. This study was aimed to assess the acute effects of radiofrequency waves emitted by a commercial smartphone on platelet function. Materials and methods Two sequential citrated blood samples were collected from 16 healthy volunteers recruited from laboratory staff. The first sample was placed in a plastic rack, 1 cm distant from a commercial smartphone receiving a 30-min call and emitting 900 MHz radiofrequency waves. The second sample was placed in another plastic rack, isolated from radiofrequency wave sources, for the same period. The platelet count and the mean platelet volume were then assessed in all blood samples, whereas platelet function was evaluated using the platelet function analyser-100 (PFA-100). Results A 30-min exposure of citrated blood to smartphone radiofrequency waves induced significant prolongation of collagen-epinephrine aggregation (median increase, 10%) and a considerable increase of mean platelet volume (median increase, 5%), whereas collagen-adenosine diphosphate aggregation and platelet count remained unchanged. Discussion This study demonstrates that smartphone radiofrequency waves induce significant perturbation of platelet structure and function, thus providing further support to concerns regarding excessive use of mobile phones. Caution should also be taken with regards to blood products containing platelets, which should be kept far away from mobile phones and smartphones throughout the production pipeline and storage period. PMID:27177410
Acute effects of 30 minutes of exposure to a smartphone call on in vitro platelet function.
Lippi, Giuseppe; Danese, Elisa; Brocco, Giorgio; Gelati, Matteo; Salvagno, Gian Luca; Montagnana, Martina
2017-05-01
Significant concerns are now regularly raised about the safety of excessive mobile phone use. This study was aimed to assess the acute effects of radiofrequency waves emitted by a commercial smartphone on platelet function. Two sequential citrated blood samples were collected from 16 healthy volunteers recruited from laboratory staff. The first sample was placed in a plastic rack, 1 cm distant from a commercial smartphone receiving a 30-min call and emitting 900 MHz radiofrequency waves. The second sample was placed in another plastic rack, isolated from radiofrequency wave sources, for the same period. The platelet count and the mean platelet volume were then assessed in all blood samples, whereas platelet function was evaluated using the platelet function analyser-100 (PFA-100). A 30-min exposure of citrated blood to smartphone radiofrequency waves induced significant prolongation of collagen-epinephrine aggregation (median increase, 10%) and a considerable increase of mean platelet volume (median increase, 5%), whereas collagen-adenosine diphosphate aggregation and platelet count remained unchanged. This study demonstrates that smartphone radiofrequency waves induce significant perturbation of platelet structure and function, thus providing further support to concerns regarding excessive use of mobile phones. Caution should also be taken with regards to blood products containing platelets, which should be kept far away from mobile phones and smartphones throughout the production pipeline and storage period.
Spore-forming organisms in platelet concentrates: a challenge in transfusion bacterial safety.
Störmer, M; Vollmer, T; Kleesiek, K; Dreier, J
2008-12-01
Bacterial detection and pathogen reduction are widely used methods of minimizing the risk of transfusion-transmitted bacterial infection. But, bacterial spores are highly resistant to chemical and physical agents. In this study, we assessed the bacterial proliferation of spore-forming organisms seeded into platelet concentrates (PCs) to demonstrate that spores can enter the vegetative state in PCs during storage. In the in vitro study, PCs were inoculated with 1-10 spores mL(-1)of Bacillus cereus (n = 1), Bacillus subtilis (n = 2) and Clostridium sporogenes (n = 2). Sampling was performed during 6-day aerobic storage at 22 degrees C. The presence of bacteria was assessed by plating culture, automated culture and real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Spores of the C. sporogenes do not enter the vegetative phase under PC storage conditions, whereas B. subtilis and B. cereus showed growth in the PC and could be detected using RT-PCR and automated culture. Depending on the species and inoculums, bacterial spores may enter the vegetative phase during PC storage and can be detected by bacterial detection methods.
Townsend, Harold E.; Barbanti, Giancarlo
1994-01-01
A nuclear fuel bundle fuel transfer system includes a transfer pool containing water at a level above a reactor core. A fuel transfer machine therein includes a carriage disposed in the transfer pool and under the water for transporting fuel bundles. The carriage is selectively movable through the water in the transfer pool and individual fuel bundles are carried vertically in the carriage. In a preferred embodiment, a first movable bridge is disposed over an upper pool containing the reactor core, and a second movable bridge is disposed over a fuel storage pool, with the transfer pool being disposed therebetween. A fuel bundle may be moved by the first bridge from the reactor core and loaded into the carriage which transports the fuel bundle to the second bridge which picks up the fuel bundle and carries it to the fuel storage pool.
Townsend, H.E.; Barbanti, G.
1994-03-01
A nuclear fuel bundle fuel transfer system includes a transfer pool containing water at a level above a reactor core. A fuel transfer machine therein includes a carriage disposed in the transfer pool and under the water for transporting fuel bundles. The carriage is selectively movable through the water in the transfer pool and individual fuel bundles are carried vertically in the carriage. In a preferred embodiment, a first movable bridge is disposed over an upper pool containing the reactor core, and a second movable bridge is disposed over a fuel storage pool, with the transfer pool being disposed therebetween. A fuel bundle may be moved by the first bridge from the reactor core and loaded into the carriage which transports the fuel bundle to the second bridge which picks up the fuel bundle and carries it to the fuel storage pool. 6 figures.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Rafique, Rashid; Xia, Jianyang; Hararuk, Oleksandra
Land models are valuable tools to understand the dynamics of global carbon (C) cycle. Various models have been developed and used for predictions of future C dynamics but uncertainties still exist. Diagnosing the models’ behaviors in terms of structures can help to narrow down the uncertainties in prediction of C dynamics. In this study three widely used land surface models, namely CSIRO’s Atmosphere Biosphere Land Exchange (CABLE) with 9 C pools, Community Land Model (version 3.5) combined with Carnegie-Ames-Stanford Approach (CLM-CASA) with 12 C pools and Community Land Model (version 4) (CLM4) with 26 C pools were driven by themore » observed meteorological forcing. The simulated C storage and residence time were used for analysis. The C storage and residence time were computed globally for all individual soil and plant pools, as well as net primary productivity (NPP) and its allocation to different plant components’ based on these models. Remotely sensed NPP and statistically derived HWSD, and GLC2000 datasets were used as a reference to evaluate the performance of these models. Results showed that CABLE exhibited better agreement with referenced C storage and residence time for plant and soil pools, as compared with CLM-CASA and CLM4. CABLE had longer bulk residence time for soil C pools and stored more C in roots, whereas, CLM-CASA and CLM4 stored more C in woody pools due to differential NPP allocation. Overall, these results indicate that the differences in C storage and residence times in three models are largely due to the differences in their fundamental structures (number of C pools), NPP allocation and C transfer rates. Our results have implications in model development and provide a general framework to explain the bias/uncertainties in simulation of C storage and residence times from the perspectives of model structures.« less
Effects of delayed laboratory processing on platelet serotonin levels.
Sanner, Jennifer E; Frazier, Lorraine; Udtha, Malini
2013-01-01
Despite the availability of established guidelines for measuring platelet serotonin, these guidelines may be difficult to follow in a hospital setting where time to processing may vary from sample to sample. The purpose of this study was to evaluate the effect of the time to processing of human blood samples on the stability of the enzyme-linked immunosorbent assay (ELISA) for the determination of platelet serotonin levels in human plasma. Human blood samples collected from a convenience sample of eight healthy volunteers were analyzed to determine platelet serotonin levels from plasma collected in ethylene diamine tetra acetic acid (EDTA) tubes and stored at 4°C for 3 hr, 5 hr, 8 hr, and 12 hr. Refrigeration storage at 4°C for 3 hr, 5 hr, 8 hr, and 12 hr altered the platelet serotonin measurement when compared to immediate processing. The bias for the samples stored at 4°C for 3 hr was 102.3 (±217.39 ng/10(9) platelets), for 5 hr was 200.1 (±132.76 ng/10(9) platelets), for 8 hr was 146.9 (±221.41 ng/10(9) platelets), and for 12 hr was -67.6 (±349.60 ng/10(9) platelets). Results from this study show that accurate measurement of platelet serotonin levels is dependent on time to processing. Researchers should therefore follow a standardized laboratory guideline for obtaining immediate platelet serotonin levels after blood sample collection.
Skripchenko, Andrey; Myrup, Andrew; Thompson-Montgomery, Dedeene; Awatefe, Helen; Wagner, Stephen J
2013-04-01
Platelet (PLT) aliquots for pediatric use have been shown to retain in vitro properties when stored in gas-impermeable syringes for up to 6 hours. As an alternative, PLT aliquots can be stored for longer periods in containers used for storage of whole blood-derived PLTs. These containers are not available separate from whole blood collection sets and PLT volumes less than 35 mL either have not been evaluated or may be unsuitable for PLT storage. Gas-permeable fluoroethylene propylene (FEP) containers have been used in the storage of cell therapy preparations and are available in multiple sizes as single containers but have not been evaluated for PLT storage. A single apheresis unit was divided on Day 3 into small aliquots with volume ranging from 20 to 60 mL, transferred using a sterile connection device, and stored for an additional 2 days either in CLX (control) or in FEP containers. PLT storage properties of PLTs stored in FEP containers were compared to those stored in CLX containers. Standard PLT in vitro assays were performed (n =6). PLT storage properties were either similar to those of CLX containers or differed by less than 20% excepting carbon dioxide levels, which varied less than 60%. Pediatric PLT aliquots of 20, 30, and 60mL transferred on Day 3 into FEP cell culture containers adequately maintain PLT properties for an additional 2days of storage. © 2012 American Association of Blood Banks.
Extended Storage of Pathogen-Reduced Platelet Concentrates (PRECON)
2017-12-01
J. Slichter, MD CONTRACTING ORGANIZATION: Bloodworks Northwest Seattle, WA 98104 REPORT DATE : December 2017 TYPE OF REPORT: Final PREPARED FOR...YOUR FORM TO THE ABOVE ADDRESS. 1. REPORT DATE December 2017 2. REPORT TYPE Final 3. DATES COVERED 26-SEP-13 to 25-SEP-17 4. TITLE AND SUBTITLE...comparator. Both the stored and the fresh platelets 4 were administered simultaneously using two different radioisotopes ( ≤15 μCi of indium for the
Leitner, G C; List, J; Horvath, M; Eichelberger, B; Panzer, S; Jilma-Stohlawetz, P
2016-01-01
Pathogen inactivation (PI) of platelet concentrates with extension of shelf life to 7 days requires the use of platelet additive solutions (PAS). We examined the quality of platelets resuspended in three different PAS stored for up to 7 days. Twelve triple adult dose platelet concentrates (PC) were collected using the TrimaAccel® collection system. Each highly concentrated product was divided into three equal parts, and the additive solutions (Composol® or SSP+® or Intersol™) were added to a final concentration of 56% PAS and 44% plasma. Samples were drawn on days 1, 5 and 7 to measure pH, glucose, lactate dehydrogenase (LDH), lactate, mean platelet volume (MPV) and the aggregation response to collagen and the thrombin receptor agonist peptide-6. Further, p-selectin expression on platelets was assessed. No statistically significant changes were observed for pH and MPV during 7 days of storage in all PAS containing PCs, whereas glucose decreased and LDH and lactate increased over time (P < 0·05). These changes were particularly evident in Intersol PCs on days 5 and 7 compared with Composol® PCs or SSP+® PCs (P < 0·05). Platelets from Intersol PCs exhibited the highest baseline activation of p-selectin and showed reduced collagen- and TRAP-6-induced aggregation. Resuspension of platelets in Intersol for 7 days results in increased platelet activation and platelet metabolism compared with SSP+® or Composol®. Further clinical studies are needed to evaluate whether the observed differences in PAS-PCs affect the recovery rate or the life span of transfused platelets. © 2015 International Society of Blood Transfusion.
Abaeva, Anastasia A.; Canault, Matthias; Kotova, Yana N.; Obydennyy, Sergey I.; Yakimenko, Alena O.; Podoplelova, Nadezhda A.; Kolyadko, Vladimir N.; Chambost, Herve; Mazurov, Aleksei V.; Ataullakhanov, Fazoil I.; Nurden, Alan T.; Alessi, Marie-Christine; Panteleev, Mikhail A.
2013-01-01
Strongly activated “coated” platelets are characterized by increased phosphatidylserine (PS) surface expression, α-granule protein retention, and lack of active integrin αIIbβ3. To study how they are incorporated into thrombi despite a lack of free activated integrin, we investigated the structure, function, and formation of the α-granule protein “coat.” Confocal microscopy revealed that fibrin(ogen) and thrombospondin colocalized as “cap,” a single patch on the PS-positive platelet surface. In aggregates, the cap was located at the point of attachment of the PS-positive platelets. Without fibrin(ogen) retention, their ability to be incorporated in aggregates was drastically reduced. The surface fibrin(ogen) was strongly decreased in the presence of a fibrin polymerization inhibitor GPRP and also in platelets from a patient with dysfibrinogenemia and a fibrinogen polymerization defect. In contrast, a fibrinogen-clotting protease ancistron increased the amount of fibrin(ogen) and thrombospondin on the surface of the PS-positive platelets stimulated with collagen-related peptide. Transglutaminases are also involved in fibrin(ogen) retention. However, platelets from patients with factor XIII deficiency had normal retention, and a pan-transglutaminase inhibitor T101 had only a modest inhibitory effect. Fibrin(ogen) retention was normal in Bernard-Soulier syndrome and kindlin-3 deficiency, but not in Glanzmann thrombasthenia lacking the platelet pool of fibrinogen and αIIbβ3. These data show that the fibrin(ogen)-covered cap, predominantly formed as a result of fibrin polymerization, is a critical mechanism that allows coated (or rather “capped”) platelets to become incorporated into thrombi despite their lack of active integrins. PMID:23995838
Muthard, Ryan W; Diamond, Scott L
2012-12-01
Blood clots form under flow during intravascular thrombosis or vessel leakage. Prevailing hemodynamics influence thrombus structure and may regulate contraction processes. A microfluidic device capable of flowing human blood over a side channel plugged with collagen (± tissue factor) was used to measure thrombus permeability (κ) and contraction at controlled transthrombus pressure drops. The collagen (κ(collagen)=1.98 × 10(-11) cm(2)) supported formation of a 20-µm thick platelet layer, which unexpectedly underwent massive platelet retraction on flow arrest. This contraction resulted in a 5.34-fold increase in permeability because of collagen restructuring. Without stopping flow, platelet deposits (no fibrin) had a permeability of κ(platelet)=5.45 × 10(-14) cm(2) and platelet-fibrin thrombi had κ(thrombus)=2.71 × 10(-14) cm(2) for ΔP=20.7 to 23.4 mm Hg, the first ever measurements for clots formed under arterial flow (1130 s(-1) wall shear rate). Platelet sensing of flow cessation triggered a 4.6- to 6.5-fold (n=3, P<0.05) increase in contraction rate, which was also observed in a rigid, impermeable parallel-plate microfluidic device. This triggered contraction was blocked by the myosin IIA inhibitor blebbistatin and by inhibitors of thromboxane A2 (TXA(2)) and ADP signaling. In addition, flow arrest triggered platelet intracellular calcium mobilization, which was blocked by TXA(2)/ADP inhibitors. As clots become occlusive or blood pools following vessel leakage, the flow diminishes, consequently allowing full platelet retraction. Flow dilution of ADP and thromboxane regulates platelet contractility with prevailing hemodynamics, a newly defined flow-sensing mechanism to regulate clot function.
21 CFR 606.121 - Container label.
Code of Federal Regulations, 2013 CFR
2013-04-01
... prepared in a system that might compromise sterility, the hour of expiration. (ii) If Source Plasma... collection date for each unit in the pool. (5) For Whole Blood, Plasma, Platelets, and partial units of Red... the container label for Source Plasma is not required to list the negative results of serological...
21 CFR 606.121 - Container label.
Code of Federal Regulations, 2014 CFR
2014-04-01
... prepared in a system that might compromise sterility, the hour of expiration. (ii) If Source Plasma... collection date for each unit in the pool. (5) For Whole Blood, Plasma, Platelets, and partial units of Red... the container label for Source Plasma is not required to list the negative results of serological...
Fuel handling system for a nuclear reactor
Saiveau, James G.; Kann, William J.; Burelbach, James P.
1986-01-01
A pool type nuclear fission reactor has a core, with a plurality of core elements and a redan which confines coolant as a hot pool at a first end of the core separated from a cold pool at a second end of the core by the redan. A fuel handling system for use with such reactors comprises a core element storage basket located outside of the redan in the cold pool. An access passage is formed in the redan with a gate for opening and closing the passage to maintain the temperature differential between the hot pool and the cold pool. A mechanism is provided for opening and closing the gate. A lifting arm is also provided for manipulating the fuel core elements through the access passage between the storage basket and the core when the redan gate is open.
Fuel handling system for a nuclear reactor
Saiveau, James G.; Kann, William J.; Burelbach, James P.
1986-12-02
A pool type nuclear fission reactor has a core, with a plurality of core elements and a redan which confines coolant as a hot pool at a first end of the core separated from a cold pool at a second end of the core by the redan. A fuel handling system for use with such reactors comprises a core element storage basket located outside of the redan in the cold pool. An access passage is formed in the redan with a gate for opening and closing the passage to maintain the temperature differential between the hot pool and the cold pool. A mechanism is provided for opening and closing the gate. A lifting arm is also provided for manipulating the fuel core elements through the access passage between the storage basket and the core when the redan gate is open.
Quebec platelet disorder: update on pathogenesis, diagnosis, and treatment.
Blavignac, Jessica; Bunimov, Natalia; Rivard, Georges E; Hayward, Catherine P M
2011-09-01
Quebec platelet disorder (QPD) is an autosomal dominant bleeding disorder associated with reduced platelet counts and a unique gain-of-function defect in fibrinolysis due to increased expression and storage of urokinase plasminogen activator (uPA) by megakaryocytes. QPD increases risks for bleeding and its key clinical feature is delayed-onset bleeding, following surgery, dental procedures or trauma, which responds only to treatment with fibrinolytic inhibitors. The genetic cause of the disorder is a tandem duplication mutation of the uPA gene, PLAU, which upregulates uPA expression in megakaryocytes by an unknown mechanism. The increased platelet stores of uPA trigger plasmin-mediated degradation of QPD α-granule proteins. The gain-of-function defect in fibrinolysis is thought to be central to the pathogenesis of QPD bleeding as the activation of QPD platelets leads to release of uPA from α-granules and accelerated clot lysis. The purpose of this review is to summarize current knowledge on QPD pathogenesis and the recommended approaches to QPD diagnosis and treatment. Thieme Medical Publishers.
NASA Astrophysics Data System (ADS)
Aubrey, D. P.; Mims, J. T.; Oswald, S. W.; Teskey, R. O.; Mitchell, R. J.
2016-12-01
Allocation of assimilated carbon to storage provides a critical carbohydrate buffer when metabolic demands exceed current photosynthetic supply; however, our process-level understanding of controls on carbon storage pools and fluxes remains relatively poor. Recent studies have shifted the paradigm from the concept that stored carbon pools are a sink of low priority that accumulate passively when photosynthetic inputs exceed demand toward the concept that these pools are active sinks of high priority. It follows that allocation toward storage—at the expense of growth—is a trait that would be under selective pressure since species that allocate toward storage should be more resilient to disturbance. Using fire-dependent longleaf pine in a series of manipulative and observational studies, we explore how stored carbon dynamics are controlled by a combination of evolutionary, physiological, and ecological pressures. Our manipulative studies revealed large stored carbon pools in roots that maintained belowground metabolism for a year after current photosynthetic supply was restricted. Likewise, the concentration of stored carbon in the smallest, most metabolically active roots was not influenced until nearly one year later. Our observational studies indicated that stored carbon pools differ among closely related species with overlapping natural distributions, but evolutionary histories of different disturbance frequencies and thus, different selective pressures on carbon storage. Our comparisons of stored carbon pools between longleaf trees growing under xeric or mesic soil moisture regimes indicated that allocation toward storage exhibits plasticity through space and time in response to both short- and long-term variations in resource availability. We expect a continuum of responses to disturbances related to ecological niche and evolutionary adaptation that influence the availability of carbohydrates for metabolic demands. We also expect a continuum in stored carbon pools and metabolic buffering capacity among species as well as spatially, temporally, and developmentally within individual species.
Duration of platelet storage and outcomes of critically ill patients.
Flint, Andrew; Aubron, Cécile; Bailey, Michael; Bellomo, Rinaldo; Pilcher, David; Cheng, Allen C; Hegarty, Colin; Reade, Michael C; McQuilten, Zoe
2017-03-01
The storage duration of platelet (PLT) units is limited to 5 to 7 days. This study investigates whether PLT storage duration is associated with patient outcomes in critically ill patients. This study was a retrospective analysis of critically ill patients admitted to the intensive care unit (ICU) of two hospitals in Australia who received one or more PLT transfusions from 2008 to 2014. Storage duration was approached in several different ways. Outcome variables were hospital mortality and ICU-acquired infection. Associations between PLT storage duration and outcomes were evaluated using multiple logistic regression and also by Cox regression. Among 2250 patients who received one or more PLT transfusions while in the ICU, the storage duration of PLTs was available for 64% of patients (1430). In-hospital mortality was 22.1% and ICU infection rate 7.2%. When comparing patients who received PLTs of a maximum storage duration of not more than 3, 4, or 5 days, there were no significant differences in baseline characteristics. After confounders were adjusted for, the storage duration of PLTs was not independently associated with mortality (4 days vs. ≤3 days, odds ratio [OR] 0.88, 95% confidence interval [CI] 0.59-1.30; 5 days vs. ≤3 days, OR 0.97, 95% CI 0.68-1.37) or infection (4 days vs. ≤3 days, OR 0.71, 95% CI 0.39-1.29; 5 days vs. ≤3 days, OR 1.11, 95% CI 0.67-1.83). Similar results were obtained regardless of how storage duration of PLTs was approached. In this large observational study in a heterogeneous ICU population, storage duration of PLTs was not associated with an increased risk of mortality or infection. © 2017 AABB.
NASA Astrophysics Data System (ADS)
Zhao, X.; Chang, Y.; Peng, F.; Wu, J.
2016-12-01
Study of solute transport in karst conduit is of great significance for prediction and prevention of groundwater pollution in southwest karst region. Solute transport in karst conduit is strongly influenced by pools which often develop along karst conduit. In order to investigate the effect of transient storage within pools on solute transport in the conduit, a pool-pipe system was built in the laboratory and some tracer tests were performed in various flow conditions to characterize the solute transport in different pool-pipe structures. The Qtracer2 program was used to obtain solute transport parameters. We used retardation coefficient R to characterize the difference between the 1-D analytical solution of the classical advection-dispersion equation and experimental results. The experimental results reveal that the concentration peak decreases with the number of pools whereas the dispersion coefficient and dispersivity increase gradually. Adding transient storage increases retardation as tailing of the breakthrough curve(BTC) is growing with the number of pools. This demonstrates that transient storage within pools is transformed to retardation. The symmetrical pool has longer tails compared to the asymmetrical pool. The concentration peak lag behind significantly due to the asymmetrical pool. A decrease in dispersivity and tailing of the BTC is observed in all pipes with the increase of flow velocities. The 1-D analytical solution of the classical advection-dispersion equation is well fitted to BTC of a single pipe in maximum flow velocity but is poorly fitted to other BTCs with appreciable tails. Therefore, it requires an appropriate model to explain tailing of the BTC. The conclusion has important significance for understanding of solute transport process in karst conduit. Future work will focus on using the appropriate model to explain tailing of the BTC.
Detrital carbon pools in temperate forests: magnitude and potential for landscape-scale assessment
John B. Bradford; Peter Weishampel; Marie-Louise Smith; Randall Kolka; Richard A. Birdsey; Scott V. Ollinger; Michael G. Ryan
2009-01-01
Reliably estimating carbon storage and cycling in detrital biomass is an obstacle to carbon accounting. We examined carbon pools and fluxes in three small temperate forest landscapes to assess the magnitude of carbon stored in detrital biomass and determine whether detrital carbon storage is related to stand structural properties (leaf area, aboveground biomass,...
Marot, Astrid; Trépo, Eric; Doerig, Christopher; Schoepfer, Alain; Moreno, Christophe; Deltenre, Pierre
2017-05-01
The 2015 Baveno VI guidelines recommend against performing upper gastrointestinal endoscopy in patients with compensated cirrhosis who have a liver stiffness <20 kPa and a platelet count >150 000/mm³ because of a low prevalence of varices at risk of bleeding in this population. The aim was to synthesize the available evidence on the usefulness of the combined use of liver stiffness and platelet count to identify patients without oesophageal varices. Meta-analysis of trials evaluating the usefulness of a given cut-off for liver stiffness and platelet count to rule out the presence of oesophageal varices. Fifteen studies were included. All studies excepting five used the Baveno VI criteria. Compared to patients with either high liver stiffness or low platelet count, those with low liver stiffness and normal platelet count had a lower risk of varices at risk of bleeding (OR=0.22, 95% CI=0.13-0.39, P<.001) with low heterogeneity between studies (I 2 =21%). They also had a lower risk of varices (OR=0.23, 95% CI=0.17-0.32, P<.001) with moderate heterogeneity between studies (I 2 =28%). In patients with low liver stiffness and normal platelet count, the pooled estimate rates for varices at risk of bleeding was 0.040 (95% CI=0.027-0.059) with low heterogeneity between studies (I 2 =3%). Patients with low liver stiffness and normal platelet count have a lower risk of varices than those with either high liver stiffness or low platelet count. Varices at risk of bleeding are found in no more than 4% of patients when liver stiffness is <20 kPa and platelet count is normal. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Insights into abnormal hemostasis in the Quebec platelet disorder from analyses of clot lysis.
Diamandis, M; Adam, F; Kahr, W H A; Wang, P; Chorneyko, K A; Arsenault, A L; Rivard, G E; Hayward, C P M
2006-05-01
The Quebec platelet disorder (QPD) is inherited and characterized by delayed-onset bleeding following hemostatic challenge. Other characteristics include increased expression and storage of active urokinase-type plasminogen activator (u-PA) in platelets in the setting of normal to increased u-PA in plasma. There is also consumption of platelet plasminogen activator inhibitor-1 and increased generation of plasmin in platelets accompanied by proteolysis of stored alpha-granule proteins, including Factor V. Although fibrinolysis has been proposed to contribute to QPD bleeding, the effects of QPD blood and platelets on clot lysis have not been evaluated. We used thromboelastography (TEG), biochemical evaluations of whole blood clot lysis, assessments of clot ultrastructure, and perfusion of blood over preformed fibrin to gain insights into the disturbed hemostasis in the QPD. Thromboelastography was not sensitive to the increased u-PA in QPD blood. However, there was abnormal plasmin generation in QPD whole blood clots, generated at low shear, with biochemical evidence of increased fibrinolysis. The incorporation of QPD platelets into a forming clot led to progressive disruption of fibrin and platelet aggregates unless drugs were added to inhibit plasmin. In whole blood perfusion studies, QPD platelets showed normal adherence to fibrin, but their adhesion was followed by accelerated fibrinolysis. The QPD is associated with "gain-of-function" abnormalities that increase the lysis of forming or preformed clots. These findings suggest accelerated fibrinolysis is an important contributor to QPD bleeding.
NASA Astrophysics Data System (ADS)
Kellman, L. M.; Gabriel, C. E.
2015-12-01
Soil organic matter (SOM) in northern forest soils is associated with a suite of minerals that can confer SOM stability, resulting in the potential for long-term storage of carbon. Increasingly, evidence is suggesting that SOM in certain mineral phases is dynamic and vulnerable to soil disturbance. The objective of this research was to investigate changes in a suite of mineral-associated pools of SOM through depth in a temperate forest soil to determine which mineral-associated carbon pools are most sensitive to forest harvesting disturbance. Sequential selective dissolutions representing increasingly stable SOM pools (soluble minerals (deionized water); humus-mineral complexes (Na-pyrophosphate); poorly crystalline minerals (HCl hydroxylamine); and crystalline secondary minerals (Na-dithionite + HCl)) of mineral soils through depth to 50 cm were carried out in podzolic soils sampled from temperate red spruce forests of contrasting stand age in Nova Scotia, Canada. Results of this analysis point to a loss of carbon from SOM within the B-horizon of the most recently harvested site from the pyrophosphate-extracted humus mineral complexed SOM, suggesting that it is this exchangeable pool that appears to be destabilized following clearcut harvesting at these study sites. This suggests that recovery from this landuse disturbance is dependent upon increasing storage of this SOM pool, and that mineral-associated pools, particularly pyrophosphate-extractable SOM, may be a useful indicator of changes to soil carbon storage following land use change.
Intravenous hemostats: challenges in translation to patients
NASA Astrophysics Data System (ADS)
Lashof-Sullivan, Margaret; Shoffstall, Andrew; Lavik, Erin
2013-10-01
Excessive bleeding and the resulting complications are a leading killer of young people globally. There are many successful methods to halt bleeding in the extremities, including compression, tourniquets, and dressings. However, current treatments for internal hemorrhage (including from head or truncal injuries), termed non-compressible bleeding, are inadequate. For these non-compressible injuries, blood transfusions are the current treatment standard. However, they must be refrigerated, may potentially transfer disease, and are of limited supply. In addition, time is of the essence for halting hemorrhage, since more than a third of civilian deaths due to hemorrhage from trauma occur before the patient even reaches the hospital. As a result, particles that can cross-link activated platelets through the glycoprotein IIb/IIIa receptor expressed on activated platelets are being investigated as an alternative treatment for non-compressible bleeding. Ideally, these particles would interact specifically with platelets to stabilize the platelet plug. Initial designs used biologically derived microparticles with red blood cell fragment or albumin cores decorated with RGD or fibrinogen, which bind to GPIIb/IIIa. More recently there has been research into the use of fully synthetic nanoparticles with liposomal or polymer cores that crosslink platelets through a targeting peptide bound to the surface. Some of the challenges for the development of these particles include appropriate sizing to prevent blocking the capillaries of the lungs, immune system evasion to prevent strong reactions and increase circulation time, and storage and resuspension so that first responders can easily use the particles. In addition, the effectiveness of the variety of animal bleeding models in predicting outcomes must be examined before test results can be fully understood. Progress has been made in the development of particles to combat hemorrhage, but issues of immune sensitivity and storage must be resolved before these types of particles can be translated for human use.
Platelet Senescence and Phosphatidylserine Exposure
Dasgupta, Swapan Kumar; Argaiz, Eduardo Rios; Chedid Mercado, Jose Emmanel; Elizondo Maul, Hector Omar; Garza, Jorge; Enriquez, Ana Bety; Abdel-Monem, Hanan; Prakasam, Anthony; Andreeff, Michael; Thiagarajan, Perumal
2010-01-01
Background The exposure of phosphatidylserine occurs during platelet activation and during in vitro storage. Phosphatidylserine exposure also occurs during apoptosis following the release of mitochondrial cytochrome c. We have examined the role of cytochrome c release, mitochondrial membrane potential (ΔΨm), and cyclophilin D (CypD) in phosphatidylserine exposure due to activation and storage. Study Design and Methods The exposure of phosphatidylserine and the loss ΔΨm were determined in a flow cytometer using FITC-lactadherin and JC-1, a lipophilic cationic reporter dye. The role of CypD was determined with cyclosporine A and CypD-deficient murine platelets. Cytochrome C induced caspase-3 and Rho associated kinase I (ROCK1) activation were determined by immunoblotting and using their inhibitors. Results Collagen and thrombin-induced exposure of phosphatidylserine was accompanied by a decrease in ΔΨm. Cyclosporin A inhibited the phosphatidylserine exposure and the loss of ΔΨm. CypD-/- mice had decreased loss of ΔΨm and impaired phosphatidylserine exposure. Collagen and thrombin did not induce the release of cytochrome c nor the activation of caspase-3 and ROCK1. In contrast, in platelets stored for more than 5 days, the phosphatidylserine exposure was associated with cytochrome c induced caspase-3 and ROCK1 activation. ABT737, a BH3 mimetic that induces mitochondrial pathway of apoptosis, induced cytochrome c release and activation of caspase-3 and ROCK1 and phosphatidylserine exposure independent of CypD. Conclusion These results show that in stored platelets cytochrome c release and the subsequent activation of caspase-3 and ROCK1 mediate phosphatidylserine exposure and it is distinct from activation-induced phosphatidylserine exposure. PMID:20456701
Carbon pools and productivity in a 1-km2 heterogeneous forest and peatland mosaic in Minnesota, USA
Peter Weishampel; Randall Kolka; Jennifer Y. King
2009-01-01
Determining the magnitude of carbon (C) storage in forests and peatlands is an important step towards predicting how regional carbon balance will respond to climate change. However, spatial heterogeneity of dominant forest and peatland cover types can inhibit accurate C storage estimates. We evaluated ecosystem C pools and productivity in the Marcell Experimental...
Carbon storage in managed forests of the northern Great Lake States
Jeanette L. Rollinger; Terry F. Strong
1996-01-01
Carbon (C) storage in forest ecosystems is a significant part of the total terrestrial C pool, and may potentially be manipulated as an important C sink. The influence of management on C pools must be understood before guidelines can be suggested for maximizing C sequestration in forests. Studies of hardwood, red pine (Pinus resinosa Ait.), aspen and...
DOE Office of Scientific and Technical Information (OSTI.GOV)
Shukla, S.D.; Hanahan, D.J.
1984-08-01
When 32P-labeled rabbits platelet were incubated with 5 X 10(-10) M 1-O-alkyl-2-acetyl-sn-glyceryl-3-phosphorylcholine (AGEPC), either in the presence or absence (0.1 mM EGTA) of added Ca2+, there was a three- to five-fold increase in the (32P)phosphatidic acid (PA) pool within 15 to 20 s. This event was followed by a gradual decrease in the (32P)PA level to near basal level in 5 min. AGEPC effected this change in (32P)PA in a characteristic dose- and time-dependent manner. Polar head group analogs of AGEPC, such as AGEDME and AGEMME, also effected an increase in PA labeling at levels comparable to those previously reportedmore » for their activity toward rabbit platelets. Other analogs, i.e., lysoGEPC and the enantiomer, sn-1-AGEPC, which are inactive toward rabbit platelets, also showed no effect on the level of (32P)PA. The finding that the PA level in rabbit platelets could be manipulated by the addition of AGEPC, without any added Ca2+, provided an excellent model system for establishing a correlation between the uptake of Ca2+, serotonin release, and PA level. Thus, PA must be regarded as a sensitive indicator of a reaction mechanism important to the platelet response to AGEPC, and could be the focal point in promoting calcium uptake during the stimulation process.« less
A novel second-generation polyolefin container for storage of single-donor apheresis platelets.
Shimizu, T; Kouketsu, K; Kamiya, T; Futagawa, H; Hirose, S
1989-01-01
A non-blow-molded LE-2 polyolefin (PO) container was developed to store single-donor apheresis platelet concentrates (PCs) processed by the Haemonetics Plasma Collecting System (PCS) at 22 degrees C for over 1 day. We molded PO containers with heat-sealing of light-weight polymer alloy films with a thickness of 0.25 mm. The film was made of polymer blends consisting of polypropylene, styrene ethylene butylene styrene-block copolymer, and ethylene ethylacrylate copolymer. The PO container with enough strength and flexibility in routine practice has 2 and 1.6 times higher oxygen and carbon dioxide gas transfer properties than standard polyvinyl chloride (PVC) plastic containers. PCs (1-1.9 x 10(11) platelets) processed from 450 ml platelet-rich plasma were stored in 0.6-liter PO containers at 22 degrees C with flatbed agitation for up to 6 days. The pH of PCs was well maintained at the mean values of 7.0 in PO containers after 6 days in the well-oxygenated condition. The energy metabolism of stored platelets was determined. Oxygen consumption rates of platelets stored in PO containers averaged 1.5 nmol/min/10(9) platelets. The rates of glucose consumption and lactate production were 0.4 and 0.8 nmol/min/10(11) platelets, respectively. The rates of adenosine triphosphate (ATP) generation of platelets, 9.7 nmol/min/10(9) platelets, in PO containers did not differ from those in the PVC containers. Aggregation responses to adenosine diphosphate and hypotonic shock response of platelets were better maintained in PO containers. The morphological changes into sphere forms with projections and the appearance of unclassified forms were more frequently observed in PO than in PVC containers.(ABSTRACT TRUNCATED AT 250 WORDS)
Pathogen-free, plasma-poor platelet lysate and expansion of human mesenchymal stem cells.
Iudicone, Paola; Fioravanti, Daniela; Bonanno, Giuseppina; Miceli, Michelina; Lavorino, Claudio; Totta, Pierangela; Frati, Luigi; Nuti, Marianna; Pierelli, Luca
2014-01-27
Supplements to support clinical-grade cultures of mesenchymal stem cells (MSC) are required to promote growth and expansion of these cells. Platelet lysate (PL) is a human blood component which may replace animal serum in MSC cultures being rich in various growth factors. Here, we describe a plasma poor pathogen-free platelet lysate obtained by pooling 12 platelet (PLT) units, to produce a standardized and safe supplement for clinical-grade expansion of MSC. PL lots were obtained by combining 2 6-unit PLT pools in additive solution (AS) following a transfusional-based procedure including pathogen inactivation (PI) by Intercept technology and 3 cycles of freezing/thawing, followed by membrane removal. Three PI-PL and 3 control PL lots were produced to compare their ability to sustain bone marrow derived MSC selection and expansion. Moreover, two further PL, subjected to PI or not, were also produced starting from the same initial PLT pools to evaluate the impact of PI on growth factor concentration and capacity to sustain cell growth. Additional PI-PL lots were used for comparison with fetal bovine serum (FBS) on MSC expansion. Immunoregulatory properties of PI-PL-generated MSC were documented in vitro by mixed lymphocyte culture (MLC) and peripheral blood mononuclear cells (PBMC) mitogen induced proliferation. PI-PL and PL control lots had similar concentrations of 4 well-described growth factors endowed with MSC stimulating ability. Initial growth and MSC expansion by PI-PL and PL controls were comparable either using different MSC populations or in head to head experiments. Moreover, PI-PL and PL control sustained similar MSC growth of frozen/thawed MSC. Multilineage differentiation of PI-derived and PI-PL-derived MSC were maintained in any MSC cultures as well as their immunoregulatory properties. Finally, no direct impact of PI on growth factor concentration and MSC growth support was observed, whereas the capacity of FBS to sustain MSC expansion in basic medium was irrelevant as compared to PL and PI-PL. The replacement of animal additives with human supplements is a basic issue in MSC ex vivo production. PI-PL represents a standardized, plasma-poor, human preparation which appears as a safe and good candidate to stimulate MSC growth in clinical-scale cultures.
Pathogen-free, plasma-poor platelet lysate and expansion of human mesenchymal stem cells
2014-01-01
Background Supplements to support clinical-grade cultures of mesenchymal stem cells (MSC) are required to promote growth and expansion of these cells. Platelet lysate (PL) is a human blood component which may replace animal serum in MSC cultures being rich in various growth factors. Here, we describe a plasma poor pathogen-free platelet lysate obtained by pooling 12 platelet (PLT) units, to produce a standardized and safe supplement for clinical-grade expansion of MSC. Methods PL lots were obtained by combining 2 6-unit PLT pools in additive solution (AS) following a transfusional-based procedure including pathogen inactivation (PI) by Intercept technology and 3 cycles of freezing/thawing, followed by membrane removal. Three PI-PL and 3 control PL lots were produced to compare their ability to sustain bone marrow derived MSC selection and expansion. Moreover, two further PL, subjected to PI or not, were also produced starting from the same initial PLT pools to evaluate the impact of PI on growth factor concentration and capacity to sustain cell growth. Additional PI-PL lots were used for comparison with fetal bovine serum (FBS) on MSC expansion. Immunoregulatory properties of PI-PL-generated MSC were documented in vitro by mixed lymphocyte culture (MLC) and peripheral blood mononuclear cells (PBMC) mitogen induced proliferation. Results PI-PL and PL control lots had similar concentrations of 4 well-described growth factors endowed with MSC stimulating ability. Initial growth and MSC expansion by PI-PL and PL controls were comparable either using different MSC populations or in head to head experiments. Moreover, PI-PL and PL control sustained similar MSC growth of frozen/thawed MSC. Multilineage differentiation of PI-derived and PI-PL-derived MSC were maintained in any MSC cultures as well as their immunoregulatory properties. Finally, no direct impact of PI on growth factor concentration and MSC growth support was observed, whereas the capacity of FBS to sustain MSC expansion in basic medium was irrelevant as compared to PL and PI-PL. Conclusion The replacement of animal additives with human supplements is a basic issue in MSC ex vivo production. PI-PL represents a standardized, plasma-poor, human preparation which appears as a safe and good candidate to stimulate MSC growth in clinical-scale cultures. PMID:24467837
Moisture sorption characteristics of freeze-dried human platelets*
Xu, Meng-jie; Chen, Guang-ming; Fan, Ju-li; Liu, Jin-hui; Xu, Xian-guo; Zhang, Shao-zhi
2011-01-01
Freeze-drying is a promising method for a long-term storage of human platelets. The moisture sorption characteristics of freeze-dried human platelets (FDHPs) were studied in this paper. The moisture sorption isotherms of FDHPs and freeze-dried lyophilization buffer (FDLB) were measured at 4, 25, and 37 °C. The experimental data were fitted to Brunauer-Emmett-Teller (BET) and Guggenheim-Anderson-de Boer (GAB) equations. There were no significant statistical differences (P>0.05) between the sorption characteristics of FDHPs and FDLB at 4 and 25 °C, while FDHPs absorbed more water at 37 °C. The net isosteric heat of sorption was derived. The heat for FDHPs showed an abnormal negative value at low moisture contents when 25 and 37 °C data were used. Dynamic sorption experiments were carried out at 25 °C with environmental water activity controlled at 0.75, 0.85, and 0.90. The moisture diffusion coefficient was fitted to be 8.24×10−12 m2/s when experimental data at initial time were used. These results would be helpful in choosing prehydration and storage condition for FDHPs. PMID:21370506
Chemical Safety Alert: Safe Storage and Handling of Swimming Pool Chemicals
Hazards of pool water treatment and maintenance chemicals (e.g., chlorine), and the protective measures pool owners should take to prevent fires, toxic vapor releases, and injuries. Triggered by improper wetting, mixing, or self-reactivity over time.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Guest, Geoffrey, E-mail: geoffrey.guest@ntnu.no; Bright, Ryan M., E-mail: ryan.m.bright@ntnu.no; Cherubini, Francesco, E-mail: francesco.cherubini@ntnu.no
2013-11-15
Temporary and permanent carbon storage from biogenic sources is seen as a way to mitigate climate change. The aim of this work is to illustrate the need to harmonize the quantification of such mitigation across all possible storage pools in the bio- and anthroposphere. We investigate nine alternative storage cases and a wide array of bio-resource pools: from annual crops, short rotation woody crops, medium rotation temperate forests, and long rotation boreal forests. For each feedstock type and biogenic carbon storage pool, we quantify the carbon cycle climate impact due to the skewed time distribution between emission and sequestration fluxesmore » in the bio- and anthroposphere. Additional consideration of the climate impact from albedo changes in forests is also illustrated for the boreal forest case. When characterizing climate impact with global warming potentials (GWP), we find a large variance in results which is attributed to different combinations of biomass storage and feedstock systems. The storage of biogenic carbon in any storage pool does not always confer climate benefits: even when biogenic carbon is stored long-term in durable product pools, the climate outcome may still be undesirable when the carbon is sourced from slow-growing biomass feedstock. For example, when biogenic carbon from Norway Spruce from Norway is stored in furniture with a mean life time of 43 years, a climate change impact of 0.08 kg CO{sub 2}eq per kg CO{sub 2} stored (100 year time horizon (TH)) would result. It was also found that when biogenic carbon is stored in a pool with negligible leakage to the atmosphere, the resulting GWP factor is not necessarily − 1 CO{sub 2}eq per kg CO{sub 2} stored. As an example, when biogenic CO{sub 2} from Norway Spruce biomass is stored in geological reservoirs with no leakage, we estimate a GWP of − 0.56 kg CO{sub 2}eq per kg CO{sub 2} stored (100 year TH) when albedo effects are also included. The large variance in GWPs across the range of resource and carbon storage options considered indicates that more accurate accounting will require case-specific factors derived following the methodological guidelines provided in this and recent manuscripts. -- Highlights: • Climate impacts of stored biogenic carbon (bio-C) are consistently quantified. • Temporary storage of bio-C does not always equate to a climate cooling impact. • 1 unit of bio-C stored over a time horizon does not always equate to − 1 unit CO{sub 2}eq. • Discrepancies of climate change impact quantification in literature are clarified.« less
Carbohydrate storage and light requirements of tropical moist and dry forest tree species.
Poorter, Lourens; Kitajima, Kaoru
2007-04-01
In many plant communities, there is a negative interspecific correlation between relative growth rates and survival of juveniles. This negative correlation is most likely caused by a trade-off between carbon allocation to growth vs. allocation to defense and storage. Nonstructural carbohydrates (NSC) stored in stems allow plants to overcome periods of stress and should enhance survival. In order to assess how species differ in carbohydrate storage in relation to juvenile light requirements, growth, and survival, we quantified NSC concentrations and pool sizes in sapling stems of 85 woody species in moist semi-evergreen and dry deciduous tropical forests in the rainy season in Bolivia. Moist forest species averaged higher NSC concentrations than dry forest species. Carbohydrate concentrations and pool sizes decreased with the light requirements of juveniles of the species in the moist forest but not in the dry forest. Combined, these results suggest that storage is especially important for species that regenerate in persistently shady habitats, as in the understory of moist evergreen forests. For moist forest species, sapling survival rates increased with NSC concentrations and pool sizes while growth rates declined with the NSC concentrations and pool sizes. No relationships were found for dry forest species. Carbon allocation to storage contributes to the growth-survival trade-off through its positive effect on survival. And, a continuum in carbon storage strategies contributes to a continuum in light requirements among species. The link between storage and light requirements is especially strong in moist evergreen forest where species sort out along a light gradient, but disappears in dry deciduous forest where light is a less limiting resource and species sort out along drought and fire gradients.
Effect of platelet-derived β-thromboglobulins on coagulation.
Egan, Karl; van Geffen, Johanna P; Ma, Hui; Kevane, Barry; Lennon, Aine; Allen, Seamus; Neary, Elaine; Parsons, Martin; Maguire, Patricia; Wynne, Kieran; O' Kennedy, Richard; Heemskerk, Johan W M; Áinle, Fionnuala Ní
2017-06-01
β-thromboglobulins are derived from the cleavage of the CXC chemokine platelet basic protein and are released in high concentrations by activated platelets. Platelet-derived β-thromboglobulins (βTG) share 70% homology with platelet factor 4 (PF4), another CXC chemokine released by activated platelets. PF4 modulates coagulation by inhibiting heparin-antithrombin interactions, promoting protein C activation, and attenuating the activity of activated protein C. In contrast, the effect of βTG on coagulation is unknown. Clotting times, thrombin generation, chromogenic clotting factor assays, and surface plasmon resonance (SPR) were used to assess the effect of purified βTG on coagulation. In normal pooled plasma, βTG shortened the lagtime and time to peak thrombin generation of tissue factor (TF)-dependent and TF-independent thrombin generation. In factor VIII and factor IX-deficient plasmas, βTG induced thrombin generation in the absence of a TF stimulus and in the presence of anti-TF and factor VIIa inhibitory antibodies. The procoagulant effect was not observed when thrombin generation was independent of factor X activation (supplementation of factor X-deficient plasma with factor Xa). Cleavage of a factor Xa-specific chromogenic substrate was observed when βTG was incubated with factor X, suggesting a direct interaction between βTG and factor X. Using SPR, βTG were found to bind to immobilised factor X in a dose dependent manner. βTG modulate coagulation in vitro via an interaction with factor X. Copyright © 2017 Elsevier Ltd. All rights reserved.
Pavlů, J; Auner, H W; Szydlo, R M; Sevillano, B; Palani, R; O'Boyle, F; Chaidos, A; Jakob, C; Kanfer, E; MacDonald, D; Milojkovic, D; Rahemtulla, A; Bradshaw, A; Olavarria, E; Apperley, J F; Pello, O M
2017-12-01
Hematopoietic precursor cells (HPC) are able to restore hematopoiesis after high-dose chemotherapy and their cryopreservation is routinely employed prior to the autologous hematopoietic cell transplantation (AHCT). Although previous studies showed feasibility of long-term HPC storage, concerns remain about possible negative effects on their potency. To study the effects of long-term cryopreservation, we compared time to neutrophil and platelet recovery in 50 patients receiving two AHCT for multiple myeloma at least 2 years apart between 2006 and 2016, using HPC obtained from one mobilization and collection attempt before the first transplant. This product was divided into equivalent fractions allowing a minimum of 2 × 10 6 CD34+ cells/kg recipient's weight. One fraction was used for the first transplant after median storage of 60 days (range, 17-165) and another fraction was used after median storage of 1448 days (range, 849-3510) at the second AHCT. Neutrophil recovery occurred at 14 days (median; range, 11-21) after the first and 13 days (10-20) after the second AHCT. Platelets recovered at a median of 16 days after both procedures. Considering other factors, such as disease status, conditioning and HPC dose, this single institution data demonstrated no reduction in the potency of HPC after long-term storage.
Robinson, Scott T; Douglas, Alison M; Chadid, Tatiana; Kuo, Katie; Rajabalan, Ajai; Li, Haiyan; Copland, Ian B; Barker, Thomas H; Galipeau, Jacques; Brewster, Luke P
2016-05-01
Mesenchymal stem cells (MSC) hold promise in promoting vascular regeneration of ischemic tissue in conditions like critical limb ischemia of the leg. However, this approach has been limited in part by poor cell retention and survival after delivery. New biomaterials offer an opportunity to localize cells to the desired tissue after delivery, but also to improve cell survival after delivery. Here we characterize the mechanical and microstructural properties of a novel hydrogel composed of pooled human platelet lysate (PL) and test its ability to promote MSC angiogenic activity using clinically relevant in vitro and in vivo models. This PL hydrogel had comparable storage and loss modulus and behaved as a viscoelastic solid similar to fibrin hydrogels despite having 1/4-1/10th the fibrin content of standard fibrin gels. Additionally, PL hydrogels enabled sustained release of endogenous PDGF-BB for up to 20days and were resistant to protease degradation. PL hydrogel stimulated pro-angiogenic activity by promoting human MSC growth and invasion in a 3D environment, and enhancing endothelial cell sprouting alone and in co-culture with MSCs. When delivered in vivo, the combination of PL and human MSCs improved local tissue perfusion after 8days compared to controls when assessed with laser Doppler perfusion imaging in a murine model of hind limb ischemia. These results support the use of a PL hydrogel as a scaffold for MSC delivery to promote vascular regeneration. Innovative strategies for improved retention and viability of mesenchymal stem cells (MSCs) are needed for cellular therapies. Human platelet lysate is a potent serum supplement that improves the expansion of MSCs. Here we characterize our novel PL hydrogel's desirable structural and biologic properties for human MSCs and endothelial cells. PL hydrogel can localize cells for retention in the desired tissue, improves cell viability, and augments MSCs' angiogenic activity. As a result of these unique traits, PL hydrogel is ideally suited to serve as a cell delivery vehicle for MSCs injected into ischemic tissues to promote vascular regeneration, as demonstrated here in a murine model of hindlimb ischemia. Published by Elsevier Ltd.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Isaka, Y.; Kimura, K.; Etani, H.
The antiplatelet effects of aspirin and ticlopidine were studied by a dual-tracer method, using indium-111 labeled platelets and technetium-99m human serum albumin, in a group of 12 patients with suspected ischemic cerebrovascular disease. The magnitude of platelet accumulation at the carotid bifurcation was expressed as the ratio of radioactivity of indium-111 platelets deposited on the vascular wall to those circulating in the blood-pool (PAI, platelet accumulation index), 48 hr after injection of labeled platelets. PAI values were measured before (baseline studies) and after the antithrombotic therapies (aspirin studies: 325 mg bid for 22.3 +/- 1.3 days, ticlopidine studies: 100 mgmore » tid for 21.8 +/- 2.1 days). At the baseline, the mean PAI value at 24 carotid bifurcations in the patient group was 15.7 +/- 15.3% (mean +/- S.D.) compared to -4.3 +/- 9.1 at 24 carotid bifurcations in 12 normal subjects (p less than 0.01). We defined the upper limit for a normal PAI (%) value to be +13.9, namely the mean PAI plus 2 SD for the carotid bifurcation in normal subjects and used this value for semiquantitative analysis. At the baseline, significant elevation of PAI (more than 13.9%; positive scintigram) was observed at 12 of 24 vessels, while 12 other regions were negative (less than 13.9%). In the lesions with positive scintigraphic results at the baseline, the mean PAI (%) value from the baseline, aspirin and ticlopidine studies was 29.5 +/- 7.0, 11.2 +/- 8.5 (p less than 0.01 versus baseline) and 21.4 +/- 21.3 (not significant from baseline), respectively.« less
Something old, something new: Why models need a multi-pool representation of storage reserves
NASA Astrophysics Data System (ADS)
Richardson, Andrew; Carbone, Mariah
2015-04-01
We know surprisingly little about processes regulating the allocation of photosynthetic assimilates to growth, storage, and other metabolic functions. Storage of nonstructural carbon (NSC, principally sugars and starch) is critically important for woody plants, because these reserves enable sessile, long-lived organisms to tolerate biotic and abiotic stress, including pests, disturbance, and drought. But, critical questions about the size and turnover of these reserves remain unanswered. Labeling studies have generally shown rapid use of new (labeled) NSC and inferred fast mixing between old and new NSC, both of which suggest quick turnover of storage reserves. However, recent studies have shown that some of the reserves stored in stem and root tissue are not only a decade old, but also still available to support new tissue growth following catastrophic disturbance. We characterized the distribution of NSC in the stemwood, branches, and roots of two temperate trees, and we used the continuous label offered by the radiocarbon (14C) bomb spike to estimate the mean age of NSC in different tissues of two temperate trees. NSC in branches and outermost stemwood growth rings had the 14C signature of the current growing season. However, NSC in older above- and below-ground tissues was enriched in 14C, indicating that it was produced from older assimilates. Radial patterns of 14C in stemwood NSC showed strong mixing of NSC across the youngest growth rings, with limited "mixing in" of younger NSC to older rings. Sugars in the outermost 5 growth rings, accounting for two-thirds of the stemwood pool, had a mean age < 1 y, whereas sugars in older growth rings had a mean age > 5 y. Thus, there is not a single, well-mixed "storage pool," and indeed "young" and "old" storage compounds appear to be physically isolated from each other. We will discuss the implications of these results for improving model representation of NSC storage and consumption by forest trees. We will suggest that there are conceptual similarities between modeling NSC pools and modeling soil C pools. We will propose future directions for modeling NSCs and also identify key questions that still need to be answered with new experimental work.
Saegusa, J; Kurikami, H; Yasuda, R; Kurihara, K; Arai, S; Kuroki, R; Matsuhashi, S; Ozawa, T; Goto, H; Takano, T; Mitamura, H; Nagano, T; Naganawa, H; Yoshida, Z; Funaki, H; Tokizawa, T; Nakayama, S
2013-03-01
Because of radioactive fallout resulting from the Fukushima Daiichi Nuclear Power Plant (NPP) accident, water discharge from many outdoor swimming pools in Fukushima was suspended out of concern that radiocesium in the pool water would flow into farmlands. The Japan Atomic Energy Agency has reviewed the existing flocculation method for decontaminating pool water and established a practical decontamination method by demonstrating the process at eight pools in Fukushima. In this method, zeolite powder and a flocculant are used for capturing radiocesium present in pool water. The supernatant is discharged if the radiocesium concentration is less than the targeted level. The radioactive residue is collected and stored in a temporary storage space. Radioactivity concentration in water is measured with a NaI(Tl) or Ge detector installed near the pool. The demonstration results showed that the pool water in which the radiocesium concentration was more than a few hundred Bq L was readily purified by the method, and the radiocesium concentration was reduced to less than 100 Bq L. The ambient dose rates around the temporary storage space were slightly elevated; however, the total increase was up to 30% of the background dose rates when the residue was shielded with sandbags.
Podda, Gian Marco; Pugliano, Mariateresa; Femia, Eti Alessandra; Mezzasoma, Anna Maria; Gresele, Paolo; Carpani, Giovanni; Cattaneo, Marco
2012-07-01
Spuriously low platelet counts (PCs) can be observed in normal blood samples anticoagulated with ethylenediamine tetra-acetic acid (EDTA)and, much less frequently, with citrate-tris-pyridossalphosphate (CPT),due to time-dependent in vitro platelet agglutination. Accuracy in PC determination is essential as PC is one of the parameters that usually guides treatment for thrombocytopenic patients. PCs of 93 thrombocy to penic patients were measured in EDTA- or CPT-anticoagulated blood samples immediately after sampling (t0) and 90 min (t90) after storage at room temperature. The presence of platelet agglutinates in blood samples was determined by examining blood smears using optical microscopy.PCs decreased at t90 with both anticoagulants. Platelet agglutinates were present at t90 in 27% of EDTA-samples vs. 2% of CPT-samples with decreased PCs (P < 0.001). Based on PCs in EDTA-samples, 15 patients (16%) shifted from a lower bleeding risk at t0 to a higher bleeding risk category at t90 (P 5 0.019), compared to 5 (5%) patients, based on PCs in CPT-samples. Therefore, time-dependent in vitro platelet agglutination in EDTA-blood samples may cause underestimation of PCs in thrombocytopenic patients, possibly leading to improper management.
16 CFR 1207.10 - Handling, storage, and marking.
Code of Federal Regulations, 2011 CFR
2011-01-01
... 16 Commercial Practices 2 2011-01-01 2011-01-01 false Handling, storage, and marking. 1207.10... REGULATIONS SAFETY STANDARD FOR SWIMMING POOL SLIDES § 1207.10 Handling, storage, and marking. (a) Marking... identification of the manufacturer. (b) Shipping, handling, and storage. The slide shall be designed, constructed...
16 CFR 1207.10 - Handling, storage, and marking.
Code of Federal Regulations, 2014 CFR
2014-01-01
... 16 Commercial Practices 2 2014-01-01 2014-01-01 false Handling, storage, and marking. 1207.10... REGULATIONS SAFETY STANDARD FOR SWIMMING POOL SLIDES § 1207.10 Handling, storage, and marking. (a) Marking... identification of the manufacturer. (b) Shipping, handling, and storage. The slide shall be designed, constructed...
16 CFR 1207.10 - Handling, storage, and marking.
Code of Federal Regulations, 2012 CFR
2012-01-01
... 16 Commercial Practices 2 2012-01-01 2012-01-01 false Handling, storage, and marking. 1207.10... REGULATIONS SAFETY STANDARD FOR SWIMMING POOL SLIDES § 1207.10 Handling, storage, and marking. (a) Marking... identification of the manufacturer. (b) Shipping, handling, and storage. The slide shall be designed, constructed...
16 CFR 1207.10 - Handling, storage, and marking.
Code of Federal Regulations, 2010 CFR
2010-01-01
... 16 Commercial Practices 2 2010-01-01 2010-01-01 false Handling, storage, and marking. 1207.10... REGULATIONS SAFETY STANDARD FOR SWIMMING POOL SLIDES § 1207.10 Handling, storage, and marking. (a) Marking... identification of the manufacturer. (b) Shipping, handling, and storage. The slide shall be designed, constructed...
Alamgir, Mohammed; Campbell, Mason J; Turton, Stephen M; Pert, Petina L; Edwards, Will; Laurance, William F
2016-07-20
Tropical forests are major contributors to the terrestrial global carbon pool, but this pool is being reduced via deforestation and forest degradation. Relatively few studies have assessed carbon storage in degraded tropical forests. We sampled 37,000 m(2) of intact rainforest, degraded rainforest and sclerophyll forest across the greater Wet Tropics bioregion of northeast Australia. We compared aboveground biomass and carbon storage of the three forest types, and the effects of forest structural attributes and environmental factors that influence carbon storage. Some degraded forests were found to store much less aboveground carbon than intact rainforests, whereas others sites had similar carbon storage to primary forest. Sclerophyll forests had lower carbon storage, comparable to the most heavily degraded rainforests. Our findings indicate that under certain situations, degraded forest may store as much carbon as intact rainforests. Strategic rehabilitation of degraded forests could enhance regional carbon storage and have positive benefits for tropical biodiversity.
Alamgir, Mohammed; Campbell, Mason J.; Turton, Stephen M.; Pert, Petina L.; Edwards, Will; Laurance, William F.
2016-01-01
Tropical forests are major contributors to the terrestrial global carbon pool, but this pool is being reduced via deforestation and forest degradation. Relatively few studies have assessed carbon storage in degraded tropical forests. We sampled 37,000 m2 of intact rainforest, degraded rainforest and sclerophyll forest across the greater Wet Tropics bioregion of northeast Australia. We compared aboveground biomass and carbon storage of the three forest types, and the effects of forest structural attributes and environmental factors that influence carbon storage. Some degraded forests were found to store much less aboveground carbon than intact rainforests, whereas others sites had similar carbon storage to primary forest. Sclerophyll forests had lower carbon storage, comparable to the most heavily degraded rainforests. Our findings indicate that under certain situations, degraded forest may store as much carbon as intact rainforests. Strategic rehabilitation of degraded forests could enhance regional carbon storage and have positive benefits for tropical biodiversity. PMID:27435389
NASA Astrophysics Data System (ADS)
Alamgir, Mohammed; Campbell, Mason J.; Turton, Stephen M.; Pert, Petina L.; Edwards, Will; Laurance, William F.
2016-07-01
Tropical forests are major contributors to the terrestrial global carbon pool, but this pool is being reduced via deforestation and forest degradation. Relatively few studies have assessed carbon storage in degraded tropical forests. We sampled 37,000 m2 of intact rainforest, degraded rainforest and sclerophyll forest across the greater Wet Tropics bioregion of northeast Australia. We compared aboveground biomass and carbon storage of the three forest types, and the effects of forest structural attributes and environmental factors that influence carbon storage. Some degraded forests were found to store much less aboveground carbon than intact rainforests, whereas others sites had similar carbon storage to primary forest. Sclerophyll forests had lower carbon storage, comparable to the most heavily degraded rainforests. Our findings indicate that under certain situations, degraded forest may store as much carbon as intact rainforests. Strategic rehabilitation of degraded forests could enhance regional carbon storage and have positive benefits for tropical biodiversity.
Fluorescence quencher improves SCANSYSTEM for rapid bacterial detection.
Schmidt, M; Hourfar, M K; Wahl, A; Nicol, S-B; Montag, T; Roth, W K; Seifried, E
2006-05-01
The optimized scansystem could detect contaminated platelet products within 24 h. However, the system's sensitivity was reduced by a high fluorescence background even in sterile samples, which led to the necessity of a well-trained staff for confirmation of microscope results. A new protocol of the optimized scansystem with the addition of a fluorescence quencher was evaluated. Pool platelet concentrates contaminated with five transfusion-relevant bacterial strains were tested in a blind study. In conjunction with new analysis software, the new quenching dye was able to reduce significantly unspecific background fluorescence. Sensitivity was best for Bacillus cereus and Escherichia coli (3 CFU/ml). The application of a fluorescence quencher enables automated discrimination of positive and negative test results in 60% of all analysed samples.
Frankincense tapping reduces the carbohydrate storage of Boswellia trees.
Mengistu, Tefera; Sterck, Frank J; Fetene, Masresha; Bongers, Frans
2013-06-01
Carbohydrates fixed by photosynthesis are stored in plant organs in the form of starch or sugars. Starch and sugars sum to the total non-structural carbohydrate pool (TNC) and may serve as intermediate pools between assimilation and utilization. We examined the impact of tapping on TNC concentrations in stem-wood, bark and root tissues of the frankincense tree (Boswellia papyrifera (Del.) Hochst) in two natural woodlands of Ethiopia. Two tapping treatments, one without tapping (control) and the other with tapping at 12 incisions, are applied on experimental trees. Trees are tapped in the leafless dry period, diminishing their carbon storage pools. If storage pools are not refilled by assimilation during the wet season, when crowns are in full leaf, tapping may deplete the carbon pool and weaken Boswellia trees. The highest soluble sugar concentrations were in the bark and the highest starch concentrations in the stem-wood. The stem-wood contains 12 times higher starch than soluble sugar concentrations. Hence, the highest TNC concentrations occurred in the stem-wood. Moreover, wood volume was larger than root or bark volumes and, as a result, more TNC was stored in the stem-wood. As predicted, tapping reduced the TNC concentrations and pool sizes in frankincense trees during the dry season. During the wet season, these carbon pools were gradually filled in tapped trees, but never to the size of non-tapped trees. We conclude that TNC is dynamic on a seasonal time scale and offers resilience against stress, highlighting its importance for tree carbon balance. But current resin tapping practices are intensive and may weaken Boswellia populations, jeopardizing future frankincense production.
Shi, Q; Schroeder, J A; Kuether, E L; Montgomery, R R
2015-07-01
Our previous studies have demonstrated that targeting FVIII expression to platelets results in FVIII storage together with von Willebrand factor (VWF) in platelet α-granules and that platelet-derived FVIII (2bF8) corrects the murine hemophilia A phenotype even in the presence of high-titer anti-FVIII inhibitory antibodies (inhibitors). To explore how VWF has an impact on platelet gene therapy for hemophilia A with inhibitors. 2bF8 transgenic mice in the FVIII(-/-) background (2bF8(tg+/-) F8(-/-) ) with varying VWF phenotypes were used in this study. Animals were analyzed by VWF ELISA, FVIII activity assay, Bethesda assay and tail clip survival test. Only 18% of 2bF8(tg+/-) F8(-/-) VWF(-/-) animals, in which VWF was deficient, survived the tail clip challenge with inhibitor titers of 3-8000 BU mL(-1) . In contrast, 82% of 2bF8(tg+/-) F8(-/-) VWF(+/+) mice, which had normal VWF levels, survived tail clipping with inhibitor titers of 10-50,000 BU mL(-1) . All 2bF8(tg+/-) F8(-/-) VWF(-/-) mice without inhibitors survived tail clipping and no VWF(-/-) F8(-/-) mice survived this challenge. Because VWF is synthesized by endothelial cells and megakaryocytes and is distributed in both plasma and platelets in peripheral blood, we further investigated the effect of each compartment of VWF on platelet-FVIII gene therapy for hemophilia A with inhibitors. In the presence of inhibitors, 42% of animals survived tail clipping in the group with plasma-VWF and 50% survived in the platelet-VWF group. VWF is essential for platelet gene therapy for hemophilia A with inhibitors. Both platelet-VWF and plasma-VWF are required for optimal platelet-derived FVIII gene therapy for hemophilia A in the presence of inhibitors. © 2015 International Society on Thrombosis and Haemostasis.
A scalable, micropore, platelet rich plasma separation device.
Dickson, Mary Nora; Amar, Levy; Hill, Michael; Schwartz, Joseph; Leonard, Edward F
2012-12-01
We have designed a novel, low energy platelet-rich-plasma (PRP) separator capable of producing 50 mL of PRP in 30 min, intended for military and emergency applications. Blood flows over a 3 mm length of sieve at high rates of shear. A plasma-platelet filtrate passes through the sieve's pores while erythrocytes remain. The filtrate is flowed over a second 3 mm length of smaller-pored sieve that withdraws plasma. Bulk blood volume is maintained by returning platelet-free plasma to the erythrocyte pool, enabling a nearly complete multi-pass platelet extraction. The total percentage of platelets extracted is:θ(T)=1-exp (-V(f)(T)Φ(P)/V) where V is the original plasma volume, V ( f )(T) is the total filtered volume, and ϕ ( P ) is platelet passage ratio (filtrate concentration/bulk average concentration) taken to be constant. Maximum θ(T) occurs at maximum V ( f )(T)× ϕ ( P ) Test microsieves, 3 mm long × 3 mm wide, were used. ϕ ( P ) values measured at various filtrate flow rates (20-100 uL/min) and utilizing various filter pore sizes (1.2-3.5 μm), was as high as 150 %. Maximum V ( f )(T)× ϕ ( P ) was achieved utilizing the 3.5 um filters at the highest flow rate, 100 uL/min. Erythrocyte leakages were always below 2,000/uL, far below the allowable limit stipulated by the American Association of Blood Banking. These data imply that a 13.7 cm(2) filter area is sufficient to achieve the target separation of 50 mL of platelet concentrate in 30 min. The filtration cartridge would consist of multiple microporous strips of 3 mm width arranged in parallel so that each element would see the conditions used in the prototype experiments presented here. Other microfiltration schemes suggest no method of scaling to practical levels.
Kwon, S Y; Kim, I S; Bae, J E; Kang, J W; Cho, Y J; Cho, N S; Lee, S W
2014-10-01
This study was conducted to evaluate the efficacy of pathogen inactivation (PI) in non-leucoreduced platelet-rich plasma-derived platelets suspended in plasma using the Mirasol PRT System and the Intercept Blood System. Platelets were pooled using the Acrodose PL system and separated into two aliquots for Mirasol and Intercept treatment. Four replicates of each viral strain were used for the evaluation. For bacteria, both low-titre (45-152 CFU/unit) inoculation and high-titre (7·34-10·18 log CFU/unit) inoculation with two replicates for each bacterial strain were used. Platelets with non-detectable bacterial growth and platelets inoculated with a low titre were stored for 5 days, and culture was performed with the BacT/ALERT system. The inactivation efficacy expressed as log reduction for Mirasol and Intercept systems for viruses was as follows: human immunodeficiency virus 1, ≥4·19 vs. ≥4·23; bovine viral diarrhoea virus, 1·83 vs. ≥6·03; pseudorabies virus, 2·73 vs. ≥5·20; hepatitis A virus, 0·62 vs. 0·76; and porcine parvovirus, 0·28 vs. 0·38. The inactivation efficacy for bacteria was as follows: Escherichia coli, 5·45 vs. ≥9·22; Staphylococcus aureus, 4·26 vs. ≥10·11; and Bacillus subtilis, 5·09 vs. ≥7·74. Postinactivation bacterial growth in platelets inoculated with a low titre of S. aureus or B. subtilis was detected only with Mirasol. Pathogen inactivation efficacy of Intercept for enveloped viruses was found to be satisfactory. Mirasol showed satisfactory inactivation efficacy for HIV-1 only. The two selected non-enveloped viruses were not inactivated by both systems. Inactivation efficacy of Intercept was more robust for all bacteria tested at high or low titres. © 2014 International Society of Blood Transfusion.
Gazit, Salomé L; Mariko, Boubacar; Thérond, Patrice; Decouture, Benoit; Xiong, Yuquan; Couty, Ludovic; Bonnin, Philippe; Baudrie, Véronique; Le Gall, Sylvain M; Dizier, Blandine; Zoghdani, Nesrine; Ransinan, Jessica; Hamilton, Justin R; Gaussem, Pascale; Tharaux, Pierre-Louis; Chun, Jerold; Coughlin, Shaun R; Bachelot-Loza, Christilla; Hla, Timothy; Ho-Tin-Noé, Benoit; Camerer, Eric
2016-09-30
Sphingosine-1-phosphate (S1P) signaling is essential for vascular development and postnatal vascular homeostasis. The relative importance of S1P sources sustaining these processes remains unclear. To address the level of redundancy in bioactive S1P provision to the developing and mature vasculature. S1P production was selectively impaired in mouse platelets, erythrocytes, endothelium, or smooth muscle cells by targeted deletion of genes encoding sphingosine kinases -1 and -2. S1P deficiency impaired aggregation and spreading of washed platelets and profoundly reduced their capacity to promote endothelial barrier function ex vivo. However, and in contrast to recent reports, neither platelets nor any other source of S1P was essential for vascular development, vascular integrity, or hemostasis/thrombosis. Yet rapid and profound depletion of plasma S1P during systemic anaphylaxis rendered both platelet- and erythrocyte-derived S1P essential for survival, with a contribution from blood endothelium observed only in the absence of circulating sources. Recovery was sensitive to aspirin in mice with but not without platelet S1P, suggesting that platelet activation and stimulus-response coupling is needed. S1P deficiency aggravated vasoplegia in this model, arguing a vital role for S1P in maintaining vascular resistance during recovery from circulatory shock. Accordingly, the S1P2 receptor mediated most of the survival benefit of S1P, whereas the endothelial S1P1 receptor was dispensable for survival despite its importance for maintaining vascular integrity. Although source redundancy normally secures essential S1P signaling in developing and mature blood vessels, profound depletion of plasma S1P renders both erythrocyte and platelet S1P pools necessary for recovery and high basal plasma S1P levels protective during anaphylactic shock. © 2016 American Heart Association, Inc.
Wagner, Stephen J; Skripchenko, Andrey; Myrup, Andrew; Thompson-Montgomery, Dedeene; Awatefe, Helen; Moroff, Gary
2010-05-01
Commercially available additive solutions (ASs) require 30% to 35% plasma for optimal storage of platelets (PLTs). PLTs suspended in M-sol, a bicarbonate-based experimental platelet additive solution (PAS), maintain in vitro PLT properties during storage with low levels of plasma (< or =5%). Four different formulations of M-sol were prepared at the optimal pH (6.1): M-sol, M-sol without calcium, M-sol without citric acid, and M-sol without calcium and citric acid. Apheresis PLT units (100% plasma) were equally divided into five 50-mL aliquots in PL732 containers, centrifuged, and resuspended to prepare units suspended in the four different PASs (95%) with 5% plasma and 1 unit in 100% plasma. Units (n = 10) were stored under standard conditions and assayed for in vitro properties on Days 1, 5, and 7. The data were analyzed by analysis of variance for repeated measures (n = 10, p < 0.001). On Day 5 of storage, PLTs suspended in the M-sol formulation containing calcium but lacking citric acid had similar pH, extent of shape change (ESC) values, and percentage of CD62-positive PLTs and greater hypotonic shock response (HSR) and percentage of discoid PLTs compared to those of PLTs suspended in 100% plasma. In contrast, PLTs suspended in the M-sol formulation lacking calcium had lesser ESC values, greater percentage of CD62-positive PLTs, and similar HSR values and percentage of discoid PLTs compared to those of PLTs suspended in 100% plasma on Day 5 (p < 0.001). Calcium plays an important role in maintaining CD62-negative PLTs and relatively high ESC in 5% plasma. The removal of citric acid from M-sol may improve PLT storage properties with low plasma levels.
16 CFR § 1207.10 - Handling, storage, and marking.
Code of Federal Regulations, 2013 CFR
2013-01-01
... 16 Commercial Practices 2 2013-01-01 2013-01-01 false Handling, storage, and marking. § 1207.10... REGULATIONS SAFETY STANDARD FOR SWIMMING POOL SLIDES § 1207.10 Handling, storage, and marking. (a) Marking... identification of the manufacturer. (b) Shipping, handling, and storage. The slide shall be designed, constructed...
LPT. Shield test facility test building interior (TAN646). Camera facing ...
LPT. Shield test facility test building interior (TAN-646). Camera facing south. Distant pool contained EBOR reactor; near pool was intended for fuel rod storage. Other post-1970 activity equipment remains in pool. INEEL negative no. HD-40-9-4 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID
Erica F. Wadl; William Lakel; Michael Aust; John Seiler
2010-01-01
Streamside management zones (SMZs) are used to protect water quality. Monitoring carbon pools and fluxes in SMZs may a good indicator of the SMZâs overall function and health. In this project we evaluated some of these pools and fluxes from three different SMZ widths (30.5, 15.3, and 7.6 m) in the Piedmont of Virginia. We quantified carbon storage in the soil (upper 10...
Fire effects on temperate forest soil C and N storage.
Nave, Lucas E; Vance, Eric D; Swanston, Christopher W; Curtis, Peter S
2011-06-01
Temperate forest soils store globally significant amounts of carbon (C) and nitrogen (N). Understanding how soil pools of these two elements change in response to disturbance and management is critical to maintaining ecosystem services such as forest productivity, greenhouse gas mitigation, and water resource protection. Fire is one of the principal disturbances acting on forest soil C and N storage and is also the subject of enormous management efforts. In the present article, we use meta-analysis to quantify fire effects on temperate forest soil C and N storage. Across a combined total of 468 soil C and N response ratios from 57 publications (concentrations and pool sizes), fire had significant overall effects on soil C (-26%) and soil N (-22%). The impacts of fire on forest floors were significantly different from its effects on mineral soils. Fires reduced forest floor C and N storage (pool sizes only) by an average of 59% and 50%, respectively, but the concentrations of these two elements did not change. Prescribed fires caused smaller reductions in forest floor C and N storage (-46% and -35%) than wildfires (-67% and -69%), and the presence of hardwoods also mitigated fire impacts. Burned forest floors recovered their C and N pools in an average of 128 and 103 years, respectively. Among mineral soils, there were no significant changes in C or N storage, but C and N concentrations declined significantly (-11% and -12%, respectively). Mineral soil C and N concentrations were significantly affected by fire type, with no change following prescribed burns, but significant reductions in response to wildfires. Geographic variation in fire effects on mineral soil C and N storage underscores the need for region-specific fire management plans, and the role of fire type in mediating C and N shifts (especially in the forest floor) indicates that averting wildfires through prescribed burning is desirable from a soils perspective.
Wyka, T.P.; Karolewski, P.; Żytkowiak, R.; Chmielarz, P.; Oleksyn, J.
2016-01-01
In evergreen plants, old leaves may contribute photosynthate to initiation of shoot growth in the spring. They might also function as storage sites for carbohydrates and nitrogen (N). We hence hypothesized that whole-plant allocation of carbohydrates and N to storage in stems and roots may be lower in evergreen than in deciduous species. We selected three species pairs consisting of an evergreen and a related deciduous species: Mahonia aquifolium (Pursh) Nutt. and Berberis vulgaris L. (Berberidaceae), Prunus laurocerasus L. and Prunus serotina Ehrh. (Rosaceae), and Viburnum rhytidophyllum Hemsl. and Viburnum lantana L. (Adoxaceae). Seedlings were grown outdoors in pots and harvested on two dates during the growing season for the determination of biomass, carbohydrate and N allocation ratios. Plant size-adjusted pools of nonstructural carbohydrates in stems and roots were lower in the evergreen species of Berberidaceae and Adoxaceae, and the slope of the carbohydrate pool vs plant biomass relationship was lower in the evergreen species of Rosaceae compared with the respective deciduous species, consistent with the leading hypothesis. Pools of N in stems and roots, however, did not vary with leaf habit. In all species, foliage contained more than half of the plant’s nonstructural carbohydrate pool and, in late summer, also more than half of the plant’s N pool, suggesting that in juvenile individuals of evergreen species, leaves may be a major storage site. Additionally, we hypothesized that concentration of defensive phenolic compounds in leaves should be higher in evergreen than in deciduous species, because the lower carbohydrate pool in stems and roots of the former restricts their capacity for regrowth following herbivory and also because of the need to protect their longer-living foliage. Our results did not support this hypothesis, suggesting that evergreen plants may rely predominantly on structural defenses. In summary, our study indicates that leaf habit has consequences for storage economics at the whole-plant level, with evergreen shrub species storing less carbohydrates (but not N) per unit plant biomass than deciduous species. PMID:26507271
Wyka, T P; Karolewski, P; Żytkowiak, R; Chmielarz, P; Oleksyn, J
2016-05-01
In evergreen plants, old leaves may contribute photosynthate to initiation of shoot growth in the spring. They might also function as storage sites for carbohydrates and nitrogen (N). We hence hypothesized that whole-plant allocation of carbohydrates and N to storage in stems and roots may be lower in evergreen than in deciduous species. We selected three species pairs consisting of an evergreen and a related deciduous species: Mahonia aquifolium (Pursh) Nutt. and Berberis vulgaris L. (Berberidaceae), Prunus laurocerasus L. and Prunus serotina Ehrh. (Rosaceae), and Viburnum rhytidophyllum Hemsl. and Viburnum lantana L. (Adoxaceae). Seedlings were grown outdoors in pots and harvested on two dates during the growing season for the determination of biomass, carbohydrate and N allocation ratios. Plant size-adjusted pools of nonstructural carbohydrates in stems and roots were lower in the evergreen species of Berberidaceae and Adoxaceae, and the slope of the carbohydrate pool vs plant biomass relationship was lower in the evergreen species of Rosaceae compared with the respective deciduous species, consistent with the leading hypothesis. Pools of N in stems and roots, however, did not vary with leaf habit. In all species, foliage contained more than half of the plant's nonstructural carbohydrate pool and, in late summer, also more than half of the plant's N pool, suggesting that in juvenile individuals of evergreen species, leaves may be a major storage site. Additionally, we hypothesized that concentration of defensive phenolic compounds in leaves should be higher in evergreen than in deciduous species, because the lower carbohydrate pool in stems and roots of the former restricts their capacity for regrowth following herbivory and also because of the need to protect their longer-living foliage. Our results did not support this hypothesis, suggesting that evergreen plants may rely predominantly on structural defenses. In summary, our study indicates that leaf habit has consequences for storage economics at the whole-plant level, with evergreen shrub species storing less carbohydrates (but not N) per unit plant biomass than deciduous species. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Luo, Yiqi; Shi, Zheng; Lu, Xingjie
Terrestrial ecosystems have absorbed roughly 30 % of anthropogenic CO 2 emissions over the past decades, but it is unclear whether this carbon (C) sink will endure into the future. Despite extensive modeling and experimental and observational studies, what fundamentally determines transient dynamics of terrestrial C storage under global change is still not very clear. Here we develop a new framework for understanding transient dynamics of terrestrial C storage through mathematical analysis and numerical experiments. Our analysis indicates that the ultimate force driving ecosystem C storage change is the C storage capacity, which is jointly determined by ecosystem C inputmore » (e.g., net primary production, NPP) and residence time. Since both C input and residence time vary with time, the C storage capacity is time-dependent and acts as a moving attractor that actual C storage chases. The rate of change in C storage is proportional to the C storage potential, which is the difference between the current storage and the storage capacity. The C storage capacity represents instantaneous responses of the land C cycle to external forcing, whereas the C storage potential represents the internal capability of the land C cycle to influence the C change trajectory in the next time step. The influence happens through redistribution of net C pool changes in a network of pools with different residence times. Moreover, this and our other studies have demonstrated that one matrix equation can replicate simulations of most land C cycle models (i.e., physical emulators). As a result, simulation outputs of those models can be placed into a three-dimensional (3-D) parameter space to measure their differences. The latter can be decomposed into traceable components to track the origins of model uncertainty. In addition, the physical emulators make data assimilation computationally feasible so that both C flux- and pool-related datasets can be used to better constrain model predictions of land C sequestration. Overall, this new mathematical framework offers new approaches to understanding, evaluating, diagnosing, and improving land C cycle models.« less
Used fuel extended storage security and safeguards by design roadmap
DOE Office of Scientific and Technical Information (OSTI.GOV)
Durbin, Samuel G.; Lindgren, Eric Richard; Jones, Robert
2016-05-01
In the United States, spent nuclear fuel (SNF) is safely and securely stored in spent fuel pools and dry storage casks. The available capacity in spent fuel pools across the nuclear fleet has nearly reached a steady state value. The excess SNF continues to be loaded in dry storage casks. Fuel is expected to remain in dry storage for periods beyond the initial dry cask certification period of 20 years. Recent licensing renewals have approved an additional 40 years. This report identifies the current requirements and evaluation techniques associated with the safeguards and security of SNF dry cask storage. Amore » set of knowledge gaps is identified in the current approaches. Finally, this roadmap identifies known knowledge gaps and provides a research path to deliver the tools and models needed to close the gaps and allow the optimization of the security and safeguards approaches for an interim spent fuel facility over the lifetime of the storage site.« less
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.
Yang, Li-Chiu; Hu, Suh-Woan; Yan, Min; Yang, Jaw-Ji; Tsou, Sing-Hua; Lin, Yuh-Yih
2015-02-01
In addition to releasing a pool of growth factors during activation, platelets have many features that indicate their role in the anti-infective host defense. The antimicrobial activities of platelet-rich plasma (PRP) and related plasma preparations against periodontal disease-associated bacteria were evaluated. Four distinct plasma fractions were extracted in the formulation used commonly in dentistry and were tested for their antibacterial properties against three periodontal bacteria: Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Fusobacterium nucleatum. The minimum inhibitory concentration of each plasma preparation was determined, and in vitro time-kill assays were used to detect their abilities to inhibit bacterial growth. Bacterial adhesion interference and the susceptibility of bacterial adherence by these plasma preparations were also conducted. All plasma preparations can inhibit bacterial growth, with PRP showing the superior activity. Bacterial growth inhibition by PRP occurred in the first 24 hours after application in the time-kill assay. PRP interfered with P. gingivalis and A. actinomycetemcomitans attachment and enhanced exfoliation of attached P. gingivalis but had no influences on F. nucleatum bacterial adherence. PRP expressed antibacterial properties, which may be attributed to platelets possessing additional antimicrobial molecules. The application of PRP on periodontal surgical sites is advisable because of its regenerative potential and its antibacterial effects.
Wang, Quan-Li; Wang, Xiao-Wei; Zhuo, Hai-Long; Shao, Chun-Yan; Wang, Jie; Wang, Hai-Ping
2013-04-01
Compared to ISBT128 code labels, radiofrequency identification (RFID) tags have incomparable advantages and gradually applied in blood management system. However, there is no global standard for the uses of RFID frequency. Even though ISBT recommended high-frequency RFID with 13.56MHz, 820- to 960-MHz ultrahigh frequency (UHF) RFID technology in many ways has even more advantages. For this reason, we studied the effect of UHF RFID tags with 820- to 960-MHz exposure on storage quality of red blood cells (RBCs) and platelets (PLTs). Thirty units of collected and prepared suspended RBCs (sRBCs) and PLTs were divided into two bags, one each for the test and control groups. The sRBCs were stored in 4±2°C refrigerator and the PLTs in a 22±2°C rocking box. The test groups were exposed to RF reader continuously during storage. Sampling at different time points and biologic changes were tested. As the extension of storage and the pH and chlorine levels in the supernatant of sRBCs were reduced, free hemoglobin, potassium, and sodium increased, but were not significant between test and control groups (p>0.05). During the storage period, the pH levels, PLT count, and PLT aggregation rate were decreased in both test and control groups, but were not significant (p>0.05). When exposed to 820- to 960-MHz RF, the biologic and biochemical indexes are not found to be exacerbated during 35 days of storage for sRBCs and 5 days for PLTs, respectively. © 2012 American Association of Blood Banks.
Mojica-Henshaw, Mariluz P; Jacobson, Pam; Morris, Julie; Kelley, Linda; Pierce, Jan; Boyer, Michael; Reems, Jo-Anna
2013-12-01
Fetal bovine serum (FBS) is commonly used as a serum supplement for culturing human mesenchymal stromal cells (hMSCs). However, human cells grown in FBS, especially for extended periods, risk potential exposure to bovine immunogenic proteins and infectious agents. To address this issue, we investigated the ability of a novel human platelet serum supplement to substitute for FBS in hMSC cultures. Platelet lysate-serum (PL-serum) was converted from platelet lysate-plasma (PL-plasma) that was manufactured from pooled platelet-rich plasma (PRP) apheresis units. Growth factor levels and the number of residual intact platelets in PL-serum and PL-plasma were compared with enzyme-linked immunosorbent assays and flow cytometry, respectively. Proliferation responses of hMSCs cultured in PL-serum, PL-plasma, or FBS were assessed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, the immunophenotype of harvested hMSCs was evaluated by flow cytometry and tri-lineage differentiation potential was evaluated by assessing adipogenic, osteogenic and chondrogenic development. Selected growth factor levels in PL-serum were not significantly different from PL-plasma (P > 0.05). hMSC cultures supplemented with PL-serum had comparable growth kinetics to PL-plasma, and hMSC yields were consistently greater than with FBS. hMSCs harvested from cultures supplemented with PL-serum, PL-plasma or FBS had similar cell surface phenotypes and maintained tri-lineage differentiation potential. PL-serum, similar to PL-plasma, can substitute for FBS in hMSC cultures. Use of PL-serum, in contrast to PL-plasma, has an added advantage of not requiring addition of a xenogeneic source of heparin, providing a completely xeno-free culture medium. Copyright © 2013 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.
Fryk, Jesse J; Marks, Denese C; Hobson-Peters, Jody; Watterson, Daniel; Hall, Roy A; Young, Paul R; Reichenberg, Stefan; Tolksdorf, Frank; Sumian, Chryslain; Gravemann, Ute; Seltsam, Axel; Faddy, Helen M
2017-11-01
Zika virus (ZIKV) has emerged as a potential threat to transfusion safety worldwide. Pathogen inactivation is one approach to manage this risk. In this study, the efficacy of the THERAFLEX UV-Platelets system and THERAFLEX MB-Plasma system to inactivate ZIKV in platelet concentrates (PCs) and plasma was investigated. PCs spiked with ZIKV were treated with the THERAFLEX UV-Platelets system at 0.05, 0.10, 0.15, and 0.20 J/cm 2 UVC. Plasma spiked with ZIKV was treated with the THERAFLEX MB-Plasma system at 20, 40, 60, and 120 J/cm 2 light at 630 nm with at least 0.8 µmol/L methylene blue (MB). Samples were taken before the first and after each illumination dose and tested for residual virus. For each system the level of viral reduction was determined. Treatment of PCs with THERAFLEX UV-Platelets system resulted in a mean of 5 log reduction in ZIKV infectivity at the standard UVC dose (0.20 J/cm 2 ), with dose dependency observed with increasing UVC dose. For plasma treated with MB and visible light, ZIKV infectivity was reduced by a mean of at least 5.68 log, with residual viral infectivity reaching the detection limit of the assay at 40 J/cm 2 (one-third the standard dose). Our study demonstrates that the THERAFLEX UV-Platelets system and THERAFLEX MB-Plasma system can reduce ZIKV infectivity in PCs and pooled plasma to the detection limit of the assays used. These findings suggest both systems have the capacity to be an effective option to manage potential ZIKV transfusion transmission risk. © 2017 AABB.
Temporal deconvolution of vascular plant signatures delivered to coastal sediments
NASA Astrophysics Data System (ADS)
Vonk, J.; Drenzek, N. J.; Hughen, K. A.; Stanley, R.; Montluçon, D. B.; McIntyre, C.; Southon, J. R.; Santos, G.; Andersson, A.; Sköld, M.; Eglinton, T. I.
2017-12-01
Presently, relatively little is known about the amount of time that lapses between the photosynthetic fixation of carbon by vascular land plants and its incorporation into the marine sedimentary record. It is clear that there are multiple potential intermediate storage pools and transport trajectories that vascular plant carbon may experience, and the age of vascular plant carbon accumulating in marine sediments will reflect these different pre-depositional histories. Here we use molecular-level radiocarbon (14C) analysis to develop down-core 14C profiles for higher plant leaf wax-derived fatty acids isolated from sediments from three sites across a 60-degrees latitudinal gradient (Cariaco Basin, Saanich Inlet, and Mackenzie Delta). The sediment profiles were used as a direct measure of the storage and transport times experienced by these biomolecular tracer compounds. Residence times are evaluated by comparing these records to the 14C history of atmospheric CO2. Using a modeling framework, we conclude that there is, in addition to a variable "young" pool, a millennial pool of compounds that consists of 49-78 % of the fractional contribution of organic carbon (OC) that exhibits variable ages for the different depositional settings. For the Mackenzie Delta sediments, we find a mean age of the millennial pool of 28 ky, suggesting pre-aging in permafrost soils, whereas the millennial pool in Saanich Inlet and Cariaco Basin sediments is younger with 7.9 and 2.4-3.2 ky, respectively, suggesting limited storage in terrestrial reservoirs. The "young" pool, conditionally defined as < 50 years showed clear annual contributions for Saanich Inlet and Mackenzie Delta sediments (24% and 16% of young pool, respectively) that can likely be explained by transport of OC from steep hillside slopes near the Saanich Inlet and annual spring flood deposition in the Mackenzie Delta. These results show that a significant fraction of vascular plant C in deltaic and marine settings undergoes pre-aging in terrestrial reservoirs. The age distribution, reflecting storage and transport times, depends on landscape-specific factors such as local topography, hydrographic characteristics, and degree of soil build-up and preservation.
Transient dynamics of terrestrial carbon storage: Mathematical foundation and numeric examples
Luo, Yiqi; Shi, Zheng; Lu, Xingjie; ...
2016-09-16
Terrestrial ecosystems absorb roughly 30% of anthropogenic CO 2 emissions since preindustrial era, but it is unclear whether this carbon (C) sink will endure into the future. Despite extensive modeling, experimental, and observational studies, what fundamentally determines transient dynamics of terrestrial C storage under climate change is still not very clear. Here we develop a new framework for understanding transient dynamics of terrestrial C storage through mathematical analysis and numerical experiments. Our analysis indicates that the ultimate force driving ecosystem C storage change is the C storage capacity, which is jointly determined by ecosystem C input (e.g., net primary production,more » NPP) and residence time. Since both C input and residence time vary with time, the C storage capacity is time-dependent and acts as a moving attractor that actual C storage chases. The rate of change in C storage is proportional to the C storage potential, the difference between the current storage and the storage capacity. The C storage capacity represents instantaneous responses of the land C cycle to external forcing, whereas the C storage potential represents the internal capability of the land C cycle to influence the C change trajectory in the next time step. The influence happens through redistribution of net C pool changes in a network of pools with different residence times. Furthermore, this and our other studies have demonstrated that one matrix equation can exactly replicate simulations of most land C cycle models (i.e., physical emulators). As a result, simulation outputs of those models can be placed into a three-dimensional (3D) parameter space to measure their differences. The latter can be decomposed into traceable components to track the origins of model uncertainty. Moreover, the emulators make data assimilation computationally feasible so that both C flux- and pool-related datasets can be used to better constrain model predictions of land C sequestration. We also propose that the C storage potential be the targeted variable for research, market trading, and government negotiation for C credits.« less
Transient dynamics of terrestrial carbon storage: Mathematical foundation and numeric examples
DOE Office of Scientific and Technical Information (OSTI.GOV)
Luo, Yiqi; Shi, Zheng; Lu, Xingjie
Terrestrial ecosystems absorb roughly 30% of anthropogenic CO 2 emissions since preindustrial era, but it is unclear whether this carbon (C) sink will endure into the future. Despite extensive modeling, experimental, and observational studies, what fundamentally determines transient dynamics of terrestrial C storage under climate change is still not very clear. Here we develop a new framework for understanding transient dynamics of terrestrial C storage through mathematical analysis and numerical experiments. Our analysis indicates that the ultimate force driving ecosystem C storage change is the C storage capacity, which is jointly determined by ecosystem C input (e.g., net primary production,more » NPP) and residence time. Since both C input and residence time vary with time, the C storage capacity is time-dependent and acts as a moving attractor that actual C storage chases. The rate of change in C storage is proportional to the C storage potential, the difference between the current storage and the storage capacity. The C storage capacity represents instantaneous responses of the land C cycle to external forcing, whereas the C storage potential represents the internal capability of the land C cycle to influence the C change trajectory in the next time step. The influence happens through redistribution of net C pool changes in a network of pools with different residence times. Furthermore, this and our other studies have demonstrated that one matrix equation can exactly replicate simulations of most land C cycle models (i.e., physical emulators). As a result, simulation outputs of those models can be placed into a three-dimensional (3D) parameter space to measure their differences. The latter can be decomposed into traceable components to track the origins of model uncertainty. Moreover, the emulators make data assimilation computationally feasible so that both C flux- and pool-related datasets can be used to better constrain model predictions of land C sequestration. We also propose that the C storage potential be the targeted variable for research, market trading, and government negotiation for C credits.« less
( sup 3 H)Dopamine uptake by platelet storage granules in schizophrenia
DOE Office of Scientific and Technical Information (OSTI.GOV)
Rabey, J.M.; Graff, E.; Oberman, Z.
1992-01-01
({sup 3}H)Dopamine (DA) uptake by platelet storage granules was determined in 26 schizophrenic male patients, paranoid type (14 acute stage; 12 in remission) and 20 age-matched, normal controls. maximum velocity (Vmax) of DA uptake was significantly higher in acute patients, than patients in remission or controls (p>0.05). The apparent Michaelis constant (kM) of DA uptake in acute patients was also significantly different from chronic patients a substantial diminution of DA uptake, while haloperidol produced a substantial diminution of DA uptake, while haloperidol (10{sup {minus}4}, 10{sup {minus}5} M) did not affect the assay. Considering that a DA disequilibrium in schizophrenia maymore » be expressed not only in the brain, but also in the periphery and that an increased amount of DA accumulated in the vesicles, implies that an increased quantity of catecholamine is available for release, our findings suggest additional evidence for the role of DA overactivity in the pathophysiology of this disorder.« less
Todd, Christopher D; Gifford, David J
2002-05-01
Following loblolly pine (Pinus taeda L.) seed germination, storage-protein breakdown in the megagametophyte and in the seedling results in a large increase in the seedling's free amino acid pool. A substantial portion of both the storage proteins and the amino acid pool is arginine, a very efficient nitrogen-storage compound. Free arginine is hydrolyzed in the seedling by the enzyme arginase (EC 3.5.3.1), which is under strong developmental control. At present, regulation of arginase in conifers is not well understood. Here we report the utilization of an in vitro culture system to address the separate impacts of the seedling and megagametophyte tissues on arginase enzyme activity, protein levels and patterns of gene expression. We also describe the generation of an anti-arginase antibody prepared from a histidine-tagged loblolly pine arginase fusion protein expressed in Escherichia coli. Our results indicate that arginase gene expression in the seedling is initiated by the seedling itself and then maintained or up-regulated by the megagametophyte. The contribution of storage-protein breakdown and the free amino acid pool, particularly arginine, in this regulation is also addressed.
Mathur, Aabhas; Chowdhury, Raquibul; Hillyer, Christopher D; Mitchell, W Beau; Shaz, Beth H
2016-12-01
Each unit of blood donated is processed and stored individually resulting in variability in the amount of red blood cells (RBCs) collected, RBC properties, and the 24-hour posttransfusion RBC survivability. As a result, each unit differs in its ability to deliver oxygen and potentially its effects on the recipient. The goal of this study was to investigate the storage of pooled RBCs from multiple donors in comparison to control standard RBC units. Two units of irradiated, leukoreduced RBCs of same ABO, D, E, C, and K antigen phenotype were collected from each of five donors using apheresis. One unit from each donor was pooled in a 2-L bag and remaining units were used as controls. After being pooled, RBCs were separated in five bags and stored at 4°C along with the controls. Quality indexes were measured on Days 2, 14, and 28 for all the units. Adenosine triphosphate assays for both pooled and controls showed a slight decrease from Day 2 to Day 28 (pooled/control from 5.22/5.24 to 4.35/4.33 µmol/g hemoglobin [Hb]). 2,3-Diphosphoglycerate was successfully rejuvenated for all RBC units on Day 28 (pooled 11.46 µmol/g Hb; control 11.86 µmol/g Hb). The results showed a nonsignificant difference between pooled and control units, with a general trend of lower standard deviation for pooled units when compared to controls. Pooled units have reduced unit-to-unit variability. Future exploration of their immunogenicity is required before using pooled units for transfusion. © 2016 AABB.
Phorbol ester stimulates calcium sequestration in saponized human platelets
DOE Office of Scientific and Technical Information (OSTI.GOV)
Yoshida, K.; Nachmias, V.T.
1987-11-25
When platelets are activated by agonists, calcium (Ca2+) is released from an intracellular storage site. Recent studies using fura-2 show that, after thrombin stimulation, the rise in free calcium is transient and returns to base-line levels in 2-3 min, while the transient following ADP stimulation lasts only 15-20 s. We reported previously that the phorbol ester 12,13-phorbol myristate acetate (PMA), added at nanomolar levels after thrombin, immediately accelerated the rate of return of calcium to the base line severalfold. In the present study, we used both intact and saponized platelets to determine whether this is due to stimulation of calciummore » sequestration. Using fura-2 and intact platelets, we found 1) that PMA stimulated the restoration of free Ca2+ levels after ADP as well as after thrombin, and 2) that H-7, an inhibitor of protein kinase C (Ca2+/phospholipid-dependent enzyme), slowed the return of Ca2+ to baseline levels. Using saponized platelets, we also found 3) that pretreatment of platelets with PMA before saponin treatment increased the ATP-dependent /sup 45/Ca2+ uptake 2-fold, with a half-maximal effect at 5 nm; 4) that most of the Ca2+ released by ionomycin or by myoinositol 1,4,5-trisphosphate; and 5) that a GTP-binding protein inhibitor, guanosine 5'-O-(2-thiodiphosphate), decreased basal or PMA-stimulated /sup 45/Ca2+ uptake in saponin-treated platelets. Our data suggest that activation of protein kinase C stimulates the sequestration of Ca2+ independently of cAMP or myoinositol 1,4,5-trisphosphate.« less
Madabhushi, Sri R; Shang, Chengwei; Dayananda, Kannayakanahalli M; Rittenhouse-Olson, Kate; Murphy, Mary; Ryan, Thomas E; Montgomery, Robert R; Neelamegham, Sriram
2012-05-17
Noncovalent association between the von Willebrand factor (VWF) propeptide (VWFpp) and mature VWF aids N-terminal multimerization and protein compartmentalization in storage granules. This association is currently thought to dissipate after secretion into blood. In the present study, we examined this proposition by quantifying the affinity and kinetics of VWFpp binding to mature VWF using surface plasmon resonance and by developing novel anti-VWF D'D3 mAbs. Our results show that the only binding site for VWFpp in mature VWF is in its D'D3 domain. At pH 6.2 and 10mM Ca(2+), conditions mimicking intracellular compartments, VWFpp-VWF binding occurs with high affinity (K(D) = 0.2nM, k(off) = 8 × 10(-5) s(-1)). Significant, albeit weaker, binding (K(D) = 25nM, k(off) = 4 × 10(-3) s(-1)) occurs under physiologic conditions of pH 7.4 and 2.5mM Ca(2+). This interaction was also observed in human plasma (K(D) = 50nM). The addition of recombinant VWFpp in both flow-chamber-based platelet adhesion assays and viscometer-based shear-induced platelet aggregation and activation studies reduced platelet adhesion and activation partially. Anti-D'D3 mAb DD3.1, which blocks VWFpp binding to VWF-D'D3, also abrogated platelet adhesion, as shown by shear-induced platelet aggregation and activation studies. Our data demonstrate that VWFpp binding to mature VWF occurs in the circulation, which can regulate the hemostatic potential of VWF by reducing VWF binding to platelet GpIbα.
Advances of blood cell-based drug delivery systems.
Sun, Yanan; Su, Jing; Liu, Geyi; Chen, Jianjun; Zhang, Xiumei; Zhang, Ran; Jiang, Minhan; Qiu, Mingfeng
2017-01-01
Blood cells, including erythrocytes, leukocytes and platelets are used as drug carriers in a wide range of applications. They have many unique advantages such as long life-span in circulation (especially erythrocytes), target release capacities (especially platelets), and natural adhesive properties (leukocytes and platelets). These properties make blood cell based delivery systems, as well as their membrane-derived carriers, far superior to other drug delivery systems. Despite the advantages, the further development of blood cell-based delivery systems was hindered by limitations in the source, storage, and mass production. To overcome these problems, synthetic biomaterials that mimic blood cell and nanocrystallization of blood cells have been developed and may represent the future direction for blood cell membrane-based delivery systems. In this paper, we review recent progress of the rising blood cell-based drug delivery systems, and also discuss their challenges and future tendency of development. Copyright © 2016. Published by Elsevier B.V.
Federal Register 2010, 2011, 2012, 2013, 2014
2010-03-11
... with zero soluble boron in the spent fuel pool, and that k eff remains less than or equal to 0.95 for the entire pool with credit for soluble boron under non-accident and accident conditions with a 95... k eff through fuel storage requirements and boron concentration controls in the spent fuel pool. The...
Lisenko, Katharina; Pavel, Petra; Kriegsmann, Mark; Bruckner, Thomas; Hillengass, Jens; Goldschmidt, Hartmut; Witzens-Harig, Mathias; Ho, Anthony D; Wuchter, Patrick
2017-04-01
Peripheral blood stem cells (PBSCs) are widely used for autologous blood stem cell transplantation (ABSCT). These cells must be stored for months or even years, usually at temperatures ≤-140°C, until their use. Although several in vitro studies on CD34 + viability and clonogenic assays of PBSCs after long-term storage have been reported, only a few publications have investigated the influence of long-term storage on in vivo hematopoietic reconstitution. In this study, we retrospectively analyzed hematopoietic recovery after storage of PBSCs via controlled-rate freezing (CRF) and cryostorage in 10% DMSO at ≤-140°C in 105 patients with multiple myeloma who received high-dose melphalan before ABSCT. Three groups of PBSC transplantation (n = 247) were delineated based on the storage period: short-term (≤12 months, n = 143), medium-term (>12 and ≤60 months, n = 75), and long-term storage (>60 months, n = 29). A neutrophil increase of ≥.5 × 10 9 /L in medium-term or long-term PBSC cryopreservation groups was observed at day 14 after ABSCT; this increase was comparable to patients who received briefly stored PBSCs (day 15). No negative effect of PBSC storage duration was observed on leucocyte or neutrophil reconstitution. Platelet reconstitutions of ≥20 × 10 9 /L and 50 × 10 9 /L were observed after median times of 10 to 11 and 13 to 14 days after ABSCT, respectively. No influence of PBSC storage duration on platelet recovery of ≥20 × 10 9 /L and ≥50 × 10 9 /L was observed in the 3 storage groups (P = .07, P = .32). The number of previous ABSCTs also had no significant impact upon hematopoietic reconstitution. In conclusion, these results indicate that long-term cryopreservation of PBSC products at vapor nitrogen temperature after CRF does not have a negative effect on hematopoietic recovery even after prolonged storage. Copyright © 2017 The American Society for Blood and Marrow Transplantation. Published by Elsevier Inc. All rights reserved.
Transient dynamics of terrestrial carbon storage: Mathematical foundation and its applications
Luo, Yiqi; Shi, Zheng; Lu, Xingjie; ...
2017-01-12
Terrestrial ecosystems have absorbed roughly 30% of anthropogenic CO 2 emissions over the past decades, but it is unclear whether this carbon (C) sink will endure into the future. Despite extensive modeling and experimental and observational studies, what fundamentally determines transient dynamics of terrestrial C storage under global change is still not very clear. Here we develop a new framework for understanding transient dynamics of terrestrial C storage through mathematical analysis and numerical experiments. Our analysis indicates that the ultimate force driving ecosystem C storage change is the C storage capacity, which is jointly determined by ecosystem C input (e.g.,more » net primary production, NPP) and residence time. Since both C input and residence time vary with time, the C storage capacity is time-dependent and acts as a moving attractor that actual C storage chases. The rate of change in C storage is proportional to the C storage potential, which is the difference between the current storage and the storage capacity. The C storage capacity represents instantaneous responses of the land C cycle to external forcing, whereas the C storage potential represents the internal capability of the land C cycle to influence the C change trajectory in the next time step. The influence happens through redistribution of net C pool changes in a network of pools with different residence times. Moreover, this and our other studies have demonstrated that one matrix equation can replicate simulations of most land C cycle models (i.e., physical emulators). As a result, simulation outputs of those models can be placed into a three-dimensional (3-D) parameter space to measure their differences. The latter can be decomposed into traceable components to track the origins of model uncertainty. In addition, the physical emulators make data assimilation computationally feasible so that both C flux- and pool-related datasets can be used to better constrain model predictions of land C sequestration. Altogether, this new mathematical framework offers new approaches to understanding, evaluating, diagnosing, and improving land C cycle models.« less
Transient dynamics of terrestrial carbon storage: Mathematical foundation and its applications
DOE Office of Scientific and Technical Information (OSTI.GOV)
Luo, Yiqi; Shi, Zheng; Lu, Xingjie
Terrestrial ecosystems have absorbed roughly 30% of anthropogenic CO 2 emissions over the past decades, but it is unclear whether this carbon (C) sink will endure into the future. Despite extensive modeling and experimental and observational studies, what fundamentally determines transient dynamics of terrestrial C storage under global change is still not very clear. Here we develop a new framework for understanding transient dynamics of terrestrial C storage through mathematical analysis and numerical experiments. Our analysis indicates that the ultimate force driving ecosystem C storage change is the C storage capacity, which is jointly determined by ecosystem C input (e.g.,more » net primary production, NPP) and residence time. Since both C input and residence time vary with time, the C storage capacity is time-dependent and acts as a moving attractor that actual C storage chases. The rate of change in C storage is proportional to the C storage potential, which is the difference between the current storage and the storage capacity. The C storage capacity represents instantaneous responses of the land C cycle to external forcing, whereas the C storage potential represents the internal capability of the land C cycle to influence the C change trajectory in the next time step. The influence happens through redistribution of net C pool changes in a network of pools with different residence times. Moreover, this and our other studies have demonstrated that one matrix equation can replicate simulations of most land C cycle models (i.e., physical emulators). As a result, simulation outputs of those models can be placed into a three-dimensional (3-D) parameter space to measure their differences. The latter can be decomposed into traceable components to track the origins of model uncertainty. In addition, the physical emulators make data assimilation computationally feasible so that both C flux- and pool-related datasets can be used to better constrain model predictions of land C sequestration. Altogether, this new mathematical framework offers new approaches to understanding, evaluating, diagnosing, and improving land C cycle models.« less
Systems biology of stored blood cells: can it help to extend the expiration date?
Paglia, Giuseppe; Palsson, Bernhard Ø; Sigurjonsson, Olafur E
2012-12-05
With increasingly stringent regulations regarding deferral and elimination of blood donors it will become increasingly important to extend the expiration date of blood components beyond the current allowed storage periods. One reason for the storage time limit for blood components is that platelets and red blood cells develop a condition called storage lesions during their storage in plastic blood containers. Systems biology provides comprehensive bio-chemical descriptions of organisms through quantitative measurements and data integration in mathematical models. The biological knowledge for a target organism can be translated in a mathematical format and used to compute physiological properties. The use of systems biology represents a concrete solution in the study of blood cell storage lesions, and it may open up new avenues towards developing better storage methods and better storage media, thereby extending the storage period of blood components. This article is part of a Special Issue entitled: Integrated omics. Copyright © 2012 Elsevier B.V. All rights reserved.
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.
2006-03-01
12,000 square feet each). An Olympic-size swimming pool (approximately 71 feet by 164 feet). Twelve covered bus stops. A skateboard park...11,644 Covered Bus Stop 100 12 1,200 Skateboard Park 10,890 1 10,890 Storage Unit 100 534 53,400 Total N/A N/A 125,134 Location to be determined...Olympic-size Swimming Pool 1 1 Covered Bus Stop 5 4 3 12 Skateboard Park 1 1 Storage Unit 0 294 160 80 534 2.6 ALTERNATIVE 1 (IMMEDIATE
NASA Astrophysics Data System (ADS)
Jordan, P. D.
2015-12-01
Annual CO2 emissions from large fixed sources in the southern San Joaquin Valley and vicinity in California are about 20 million metric tons per year (MMT/Y). Cumulative net fluid production due to oil and gas extracted from below the minimum depth for geologic carbon storage (taken as 1,500 m) was 1.4 billion m3 at reservoir conditions as of 2010. At an average CO2 storage density of 0.5 metric tons per m3, this implies 35 years of storage capacity at current emission rates just to refill the vacated volume, neglecting possible reservoir consolidation. However, the production occurred from over 300 pools. The production rate relative to average pressure decline in the more productive pools analyzed suggests they could receive about 2 MMT/Y raising the field average pressure to nearly the fracturing pressure. This would require well fields as extensive as those used for production, instead of the single to few wells per project typically envisioned. Even then, the actual allowable injection rate to the larger pools would be less than 2 MMT/Y in order to keep pressures at the injection well below the fracture pressure. This implies storing 20 MMT/Y would require developing storage operations in tens of pools with hundreds, if not over a thousand, wells. This utilization of one of the basins with the most storage capacity in the state would result in reducing the state's fixed source emissions by only one eighth relative to current emissions. The number of fields and wells involved in achieving this suggests a different strategy might provide more capacity at similar cost. Specifically, staging wells that initially produce water in the vicinity of fewer injection wells could result in both more storage. This water could be directed to a shallower zone, or supplied to the surface at a similar cost. The commencement of ocean water desalination in the state indicates the economics of water supply might support treating this water for beneficial use, particularly if it has a lower salinity than sea water.
Fission matrix-based Monte Carlo criticality analysis of fuel storage pools
DOE Office of Scientific and Technical Information (OSTI.GOV)
Farlotti, M.; Ecole Polytechnique, Palaiseau, F 91128; Larsen, E. W.
2013-07-01
Standard Monte Carlo transport procedures experience difficulties in solving criticality problems in fuel storage pools. Because of the strong neutron absorption between fuel assemblies, source convergence can be very slow, leading to incorrect estimates of the eigenvalue and the eigenfunction. This study examines an alternative fission matrix-based Monte Carlo transport method that takes advantage of the geometry of a storage pool to overcome this difficulty. The method uses Monte Carlo transport to build (essentially) a fission matrix, which is then used to calculate the criticality and the critical flux. This method was tested using a test code on a simplemore » problem containing 8 assemblies in a square pool. The standard Monte Carlo method gave the expected eigenfunction in 5 cases out of 10, while the fission matrix method gave the expected eigenfunction in all 10 cases. In addition, the fission matrix method provides an estimate of the error in the eigenvalue and the eigenfunction, and it allows the user to control this error by running an adequate number of cycles. Because of these advantages, the fission matrix method yields a higher confidence in the results than standard Monte Carlo. We also discuss potential improvements of the method, including the potential for variance reduction techniques. (authors)« less
Koltun, G.F.
2009-01-01
This report describes the results of a study to determine frequency characteristics of postregulation annual peak flows at streamflow-gaging stations at or near the Lockington, Taylorsville, Englewood, Huffman, and Germantown dry dams in the Miami Conservancy District flood-protection system (southwestern Ohio) and five other streamflow-gaging stations in the Great Miami River Basin further downstream from one or more of the dams. In addition, this report describes frequency characteristics of annual peak elevations of the dry-dam pools. In most cases, log-Pearson Type III distributions were fit to postregulation annual peak-flow values through 2007 (the most recent year of published peak-flow values at the time of this analysis) and annual peak dam-pool storage values for the period 1922-2008 to determine peaks with recurrence intervals of 2, 5, 10, 25, 50, 100, 200, and 500 years. For one streamflow-gaging station (03272100) with a short period of record, frequency characteristics were estimated by means of a process involving interpolation of peak-flow yields determined for an upstream and downstream gage. Once storages had been estimated for the various recurrence intervals, corresponding dam-pool elevations were determined from elevation-storage ratings provided by the Miami Conservancy District.
pH value promotes growth of Staphylococcus epidermidis in platelet concentrates.
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.
Platelet concentrates prepared after a 20- to 24-hour hold of the whole blood at 22°C.
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.
K. J. McFarlane; S. H. Schoenholtz; R. F. Powers
2009-01-01
Belowground C and N storage is important in maintaining forestproductivity and to CO2 sequestration. How these pools respondto management is poorly understood. We investigated effectsof repeated applications of complete fertilizer and competing...
Kathleen A. Dwire; Ellen E. Wohl; Nicholas A. Sutfin; Roberto A. Bazan; Lina Polvi-Pilgrim
2012-01-01
Headwaters are known to be important in the global carbon cycle, yet few studies have investigated carbon (C) pools along stream-riparian corridors. To better understand the spatial distribution of C storage in headwater fluvial networks, we estimated above- and below-ground C pools in 100-m-long reaches in six different valley types in Rocky Mountain National Park,...
Benefits of tree mixes in carbon plantings
NASA Astrophysics Data System (ADS)
Hulvey, Kristin B.; Hobbs, Richard J.; Standish, Rachel J.; Lindenmayer, David B.; Lach, Lori; Perring, Michael P.
2013-10-01
Increasingly governments and the private sector are using planted forests to offset carbon emissions. Few studies, however, examine how tree diversity -- defined here as species richness and/or stand composition -- affects carbon storage in these plantings. Using aboveground tree biomass as a proxy for carbon storage, we used meta-analysis to compare carbon storage in tree mixtures with monoculture plantings. Tree mixes stored at least as much carbon as monocultures consisting of the mixture's most productive species and at times outperformed monoculture plantings. In mixed-species stands, individual species, and in particular nitrogen-fixing trees, increased stand biomass. Further motivations for incorporating tree richness into planted forests include the contribution of diversity to total forest carbon-pool development, carbon-pool stability and the provision of extra ecosystem services. Our findings suggest a two-pronged strategy for designing carbon plantings including: (1) increased tree species richness; and (2) the addition of species that contribute to carbon storage and other target functions.
Distribution and dosimetry of In-111 labeled leukocytes and platelets in humans
DOE Office of Scientific and Technical Information (OSTI.GOV)
Goodwin, D.A.; Finston, R.A.; Smith, S.I.
1981-06-01
The distribution of In-111 labeled leukocytes and platelets was studied by whole body gamma camera imaging in patients. Images were made approximately one hour and 24 hours after IV injection, and stored in digital form in computer memory. Estimates of the quantitative organ distribution were made from the geometric mean of the anterior and posterior region of interest counts after suitable background subtraction. Nearly quantitative retention of cell activity was observed with little or no excretion seen in either gut or kidneys. Mixed leukocytes distributed in spleen, liver and bone marrow in decreasing order of concentration, similar at both times,more » with a transient lung uptake noted at the one hour time only. The dose from 0.5 mCi In-111-WBC's was: liver, 1.4 rad; spleen, 8.5 rad; marrow, 2.3 rad. Lymphocytes had similar distribution with the addition of inguinal and cervical lymph nodes. The dose from 0.5 mCi In-111-lymphocytes was: liver, 0.8 rad; spleen, 6.7 rad; marrow and lymphatic tissue, 1.4 rad. Platelets distributed primarily in the blood pool with most of the remainder concentrating in the spleen, with a small amount in the penis. The dose from 0.5 mCi of In-111-platelets was: liver, 3.2 rad; spleen, 8.6 rad; and whole body, 0.3 rad.« less
Burnouf, Thierry; Goubran, Hadi Alphonse; Chou, Ming-Li; Devos, David; Radosevic, Mirjana
2014-07-01
There is increasing research on and clinical interest in the physiological role played by platelet microparticles (PMPs). PMPs are 0.1-1-μm fragments shed from plasma membranes of platelets that are undergoing activation, stress, or apoptosis. They have a phospholipid-based structure and express functional receptors from platelet membranes. As they are the most abundant microparticles in the blood and they express the procoagulant phosphatidylserine, PMPs likely complement, if not amplify, the functions of platelets in hemostasis, thrombosis, cancer, and inflammation, but also act as promoters of tissue regeneration. Their size and structure make them instrumental in platelet-cell communications as a delivery tool of platelet-borne bioactive molecules including growth factors, other signaling molecules and micro (mi)RNA. PMPs can therefore be a pathophysiological threat or benefit to the cellular environment when interacting with the blood vasculature. There is also increasing evidence that PMP generation is triggered during blood collection, separation into components, and storage, a phenomenon potentially leading to thrombotic and inflammatory side effects in transfused patients. Evaluating PMPs requires strict pre-analytical and analytical procedures to avoid artifactual generation and ensure accurate assessment of the number, size repartitioning, and functional properties. This review describes the physical and functional methods developed for analyzing and quantifying PMPs. It then presents the functional roles of PMPs as markers or triggers of diseases like thrombosis, atherosclerosis, and cancer, and discusses the possible detrimental immunological impact of their generation in blood components. Finally we review the potential function of PMPs in tissue regeneration and the prospects for their use in therapeutic strategies for human health. Copyright © 2014 Elsevier Ltd. All rights reserved.
Proliferation resistance assessment of various methods of spent nuclear fuel storage and disposal
NASA Astrophysics Data System (ADS)
Kollar, Lenka
Many countries are planning to build or already are building new nuclear power plants to match their growing energy needs. Since all nuclear power plants handle nuclear materials that could potentially be converted and used for nuclear weapons, they each present a nuclear proliferation risk. Spent nuclear fuel presents the largest build-up of nuclear material at a power plant. This is a proliferation risk because spent fuel contains plutonium that can be chemically separated and used for a nuclear weapon. The International Atomic Energy Agency (IAEA) safeguards spent fuel in all non-nuclear weapons states that are party to the Non-Proliferation Treaty. Various safeguards methods are in use at nuclear power plants and research is underway to develop safeguards methods for spent fuel in centralized storage or underground storage and disposal. Each method of spent fuel storage presents different proliferation risks due to the nature of the storage method and the safeguards techniques that are utilized. Previous proliferation resistance and proliferation risk assessments have mainly compared nuclear material through the whole fuel cycle and not specifically focused on spent fuel storage. This project evaluates the proliferation resistance of the three main types of spent fuel storage: spent fuel pool, dry cask storage, and geological repository. The proliferation resistance assessment methodology that is used in this project is adopted from previous work and altered to be applicable to spent fuel storage. The assessment methodology utilizes various intrinsic and extrinsic proliferation-resistant attributes for each spent fuel storage type. These attributes are used to calculate a total proliferation resistant (PR) value. The maximum PR value is 1.00 and a greater number means that the facility is more proliferation resistant. Current data for spent fuel storage in the United States and around the world was collected. The PR values obtained from this data are 0.49 for the spent fuel pool, 0.42 for dry cask storage, 0.36 for the operating geological repository, and 0.28 for the closed geological repository. Therefore, the spent fuel pool is currently the most proliferation resistant method for storing spent fuel. The extrinsic attributes, mainly involving safeguards measures, affect the total PR value the most. As a result, several recommendations are made to improve the proliferation resistance of spent fuel. These recommendations include employing more advanced safeguards measures, such as verification techniques and remote monitoring, for dry cask storage and the geological repository. Dry cask storage facilities should also be located at the plant and in a secure building to minimize the proliferation risk. Finally, the cost-benefit analysis of increased safeguards needs to be considered. Taking these recommendations into account, the PR values of dry cask storage and the closed geological would be significantly increased, to 0.57 and 0.51, respectively. As a result, with increased safeguards to the safeguards level of the spent fuel pool, dry cask storage would be the most proliferation resistant method to store spent fuel. Therefore, the IAEA should continue to develop remote monitoring and cask storage verification techniques in order to improve the proliferation resistance of spent fuel.
Vamvakas, Eleftherios C
2009-12-01
Risks of transfusion-transmitted infections (TTIs), transfusion-associated sepsis (TAS), and transfusion-related acute lung injury (TRALI) were compared between pooled whole blood-derived (PWBD) and single-donor platelets (PLTs) transfused in the United States. The literature was searched for estimates of the risk of TTIs and TAS and of the effect on bacterial contamination of PLTs of process improvements, bacterial culture, and surrogate methods to detect bacteria. Seven studies published between January 2005 and December 2008 and comparing bacterial contamination frequency between PWBD and single-donor PLTs after implementing bacterial culture testing of both components were subjected to meta-analysis. The three retrieved studies diagnosing TRALI based on the 2004 consensus definition in settings transfusing both PWBD and single-donor PLTs were not amenable to meta-analysis and were assessed qualitatively. Under a best-case scenario, if 100% (from the current 12.5%) of PLT doses were provided as PWBD PLTs, the number of additional transmissions of human immunodeficiency virus, hepatitis C virus, hepatitis B virus, bacteria, or a novel pathogen annually could be 1.2, 1.3, 9.0, 105.3, or 69.2 to 252.6, respectively. Compared with single-donor PLTs, US PLT pools of five concentrates have a 5.6-fold higher risk of bacterial contamination (summary odds ratio, 5.58; 95% confidence interval, 2.60-11.98; p < 0.05). The three studies that diagnosed TRALI based on the consensus definition did not demonstrate a difference in risk between PWBD and single-donor PLTs. TTIs and TAS determine the relative safety of PWBD versus single-donor PLTs. The available limited data do not support a higher risk of TRALI from single-donor (compared with PWBD) PLTs.
Lip and tooth injuries at public swimming pools in Austria.
Lechner, Katharina; Connert, Thomas; Kühl, Sebastian; Filippi, Andreas
2017-06-01
There is an increased risk of orofacial injuries in swimming pool facilities. Nevertheless, only a few studies have addressed this issue. The aim of this study was to identify the frequency of lip and tooth injuries at public swimming pools in Austria. A further aim was to examine which gender and age groups were affected, where and why these injuries occurred, and whether pool attendants had sufficient knowledge of dental first-aid measures. A total of 764 pool attendants in Austria were contacted by telephone and 689 participated in the study (90.2%). The attendants were interviewed retrospectively about accident occurrences in 2014 by a standardized questionnaire. Responses to the provision of first aid and choice of storage medium for avulsed teeth were subsequently evaluated. The frequency of lip injuries was 19.0%, and tooth injuries were 11.3%. Male bathers (P < .05) and children under 12 years (P < .001) most frequently suffered injuries. The waterslide was the most common accident site. The most common cause of lip injuries was slipping on wet surfaces (39.0%), and for tooth injuries it was collisions with other persons or objects (each 28.1%). The pool attendants' responses were predominantly good or sufficient on first aid, with the exception of what storage medium to choose. Tooth rescue boxes were available in only 8.6% of all pool facilities. Orofacial injuries are a frequently occurring problem in swimming pool facilities. The pool attendants' knowledge on first-aid care of tooth injuries could still be improved. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Elkin, L L; Harden, D G; Saldanha, S; Ferguson, H; Cheney, D L; Pieniazek, S N; Maloney, D P; Zewinski, J; O'Connell, J; Banks, M
2015-06-01
Compound pooling, or multiplexing more than one compound per well during primary high-throughput screening (HTS), is a controversial approach with a long history of limited success. Many issues with this approach likely arise from long-term storage of library plates containing complex mixtures of compounds at high concentrations. Due to the historical difficulties with using multiplexed library plates, primary HTS often uses a one-compound-one-well approach. However, as compound collections grow, innovative strategies are required to increase the capacity of primary screening campaigns. Toward this goal, we have developed a novel compound pooling method that increases screening capacity without compromising data quality. This method circumvents issues related to the long-term storage of complex compound mixtures by using acoustic dispensing to enable "just-in-time" compound pooling directly in the assay well immediately prior to assay. Using this method, we can pool two compounds per well, effectively doubling the capacity of a primary screen. Here, we present data from pilot studies using just-in-time pooling, as well as data from a large >2-million-compound screen using this approach. These data suggest that, for many targets, this method can be used to vastly increase screening capacity without significant reduction in the ability to detect screening hits. © 2015 Society for Laboratory Automation and Screening.
Steam-cooking rapidly destroys and reverses onion-induced antiplatelet activity
2012-01-01
Background Foods in the diet that can aid in the prevention of diseases are of major interest. Onions are key ingredients in many cuisines around the world and moreover, onion demand has trended higher over the past three decades. An important pharmacological aspect of onion is the ability to inhibit platelet aggregation. Raw onions inhibit platelet aggregation; however, when onions are boiled or heated, antiplatelet activity may be abolished. Methods Onion quarters were steamed for 0, 1, 3, 6, 10, and 15 min. The in vitro antiplatelet activity of a yellow hybrid storage onion was examined at these times on the blood of 12 human subjects using in vitro whole blood aggregometry. Results Contrary to findings reported for boiling, antiplatelet activity was destroyed between 3 and 6 min of steaming, and at 10 min of steaming, cooked onions stimulated platelet activity. Extracts from cooked onion had the potential to reverse the inhibitory effect on blood platelets by 25%. Responses were consistent across all donors. Total polyphenolic concentration and soluble solids were not affected by steaming time. Conclusions The potential value of cooked onion preparations may result in destruction or reversal of antiplatelet activity, without affecting the polyphenolic concentration. PMID:22992282
Steam-cooking rapidly destroys and reverses onion-induced antiplatelet activity.
Hansen, Emilie A; Folts, John D; Goldman, Irwin L
2012-09-20
Foods in the diet that can aid in the prevention of diseases are of major interest. Onions are key ingredients in many cuisines around the world and moreover, onion demand has trended higher over the past three decades. An important pharmacological aspect of onion is the ability to inhibit platelet aggregation. Raw onions inhibit platelet aggregation; however, when onions are boiled or heated, antiplatelet activity may be abolished. Onion quarters were steamed for 0, 1, 3, 6, 10, and 15 min. The in vitro antiplatelet activity of a yellow hybrid storage onion was examined at these times on the blood of 12 human subjects using in vitro whole blood aggregometry. Contrary to findings reported for boiling, antiplatelet activity was destroyed between 3 and 6 min of steaming, and at 10 min of steaming, cooked onions stimulated platelet activity. Extracts from cooked onion had the potential to reverse the inhibitory effect on blood platelets by 25%. Responses were consistent across all donors. Total polyphenolic concentration and soluble solids were not affected by steaming time. The potential value of cooked onion preparations may result in destruction or reversal of antiplatelet activity, without affecting the polyphenolic concentration.
Pohler, Petra; Müller, Meike; Winkler, Carla; Schaudien, Dirk; Sewald, Katherina; Müller, Thomas H; Seltsam, Axel
2015-02-01
Residual white blood cells (WBCs) in cellular blood components induce a variety of adverse immune events, including nonhemolytic febrile transfusion reactions, alloimmunization to HLA antigens, and transfusion-associated graft-versus-host disease (TA-GVHD). Pathogen reduction (PR) methods such as the ultraviolet C (UVC) light-based THERAFLEX UV-Platelets system were developed to reduce the risk of transfusion-transmitted infection. As UVC light targets nucleic acids, it interferes with the replication of both pathogens and WBCs. This preclinical study aimed to evaluate the ability of UVC light to inactivate contaminating WBCs in platelet concentrates (PCs). The in vitro and in vivo function of WBCs from UVC-treated PCs was compared to that of WBCs from gamma-irradiated and untreated PCs by measuring cell viability, proliferation, cytokine secretion, antigen presentation in vitro, and xenogeneic GVHD responses in a humanized mouse model. UVC light was at least as effective as gamma irradiation in preventing GVHD in the mouse model. It was more effective in suppressing T-cell proliferation (>5-log reduction in the limiting dilution assay), cytokine secretion, and antigen presentation than gamma irradiation. The THERAFLEX UV-Platelets (MacoPharma) PR system can substitute gamma irradiation for TA-GVHD prophylaxis in platelet (PLT) transfusion. Moreover, UVC treatment achieves suppression of antigen presentation and inhibition of cytokine accumulation during storage of PCs, which has potential benefits for transfusion recipients. © 2014 AABB.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Dawson, D.M.; Guerra, G.; Neider, T.
1995-12-01
This report describes the system developed by EPRI/DOE for the dry transfer of spent fuel assemblies outside the reactor spent fuel pool. The system is designed to allow spent fuel assemblies to be removed from a spent fuel pool in a small cask, transported to the transfer facility, and transferred to a larger cask, either for off-site transportation or on-site storage. With design modifications, this design is capable of transferring single spent fuel assemblies from dry storage casks to transportation casks or visa versa. One incentive for the development of this design is that utilities with limited lifting capacity ormore » other physical or regulatory constraints are limited in their ability to utilize the current, more efficient transportation and storage cask designs. In addition, DOE, in planning to develop and implement the multi-purpose canister (MPC) system for the Civilian Radioactive Waste Management System, included the concept of an on-site dry transfer system to support the implementation of the MPC system at reactors with limitations that preclude the handling of the MPC system transfer casks. This Dry Transfer System can also be used at reactors wi decommissioned spent fuel pools and fuel in dry storage in non-MPC systems to transfer fuel into transportation casks. It can also be used at off-reactor site interim storage facilities for the same purpose.« less
Human Milk Adrenomedullin Is Unstable During Cold Storage at 4°C.
Peila, Chiara; Coscia, Alessandra; Bertino, Enrico; Li Volti, Giovanni; Galvano, Fabio; Barbagallo, Ignazio; Gazzolo, Diego
2017-11-01
Under some circumstances human milk (HM) extraction and refrigerated storage may be necessary. Depending on the length and on the type of cold storage, milk may lose some important properties, but current advices on safe HM storage are discordant. Moreover until now no data in literature were present on the effect of prolonged cold storage on biologically active components of the HM such as adrenomedullin (AM). This important peptide is involved in response to hypoxia and inflammation, associated with neovascularization, in several tissues. The aim is to evaluate: (a) the presence of AM in preterm and term HM and (b) the concentration of AM in refrigerated milk at 4°C at 24-hour intervals, up to 96 hours of storage. The experiment was repeated four times. Immediately after collection, each HM sample deriving from each mother was divided into two parts as follows: "Pool" line and "Single Mother" line. One part (Pool line) was pooled and then divided into five aliquots. The other part (Single Mother line) was divided into five aliquots. From each line, one aliquot was analyzed within 3 hours, while the others were stored in the refrigerator for 24, 48, 72, and 96 hours, respectively, and then analyzed. AM levels were determined using a specific ELISA test. AM was detectable in all samples. Its concentration was significantly higher in preterm milk with respect to term milk (p < 0.05). Significant differences were observed during the cold storage: the AM levels decreased steadily during the storage and the remaining concentration at 96 hours is ∼2%. This study provides evidences regarding the presence of AM in HM, regardless of the gestational age. In particular, the refrigeration of fresh HM in controlled conditions significantly affected its bioactivity and nutritional quality related with AM, already at 24 hours.
NASA Astrophysics Data System (ADS)
Vourlitis, G. L.; Hentz, C. S.
2015-12-01
Mediterranean-type shublands are subject to periodic fire and high levels of atmospheric nitrogen (N) deposition. Little is known how N inputs interact with post-fire secondary succession to affect ecosystem carbon (C) and N storage and cycling. Thus, a field experiment was conducted in a chaparral stand located in NE San Diego County, USA that burned during a wildfire in July 2003 to test the hypotheses that rates of C and N storage would significantly increase in response to experimental N addition. The experimental layout consists of a randomized design where four-10 x 10 m plots received 5 gN m-2 (added N) in the fall of each year since 2003 and four-10 x 10 m plots served as un-manipulated controls. Aboveground biomass C and N pools and fluxes, including biomass and litter C and N pool size, litter production, net primary production (NPP), N uptake, and litter C and N mineralization were measured seasonally (every 3 months) for a period of 10 years. Belowground surface (0-10 cm) soil extractable N, pH, and total soil N and C pools and surface root biomass C and N pools were also measured seasonally for a period of 10 years, while N losses from leaching were measured over a shorted (8 year) period of time. Added N led to a rapid increase in soil extractable N and a decline in soil pH; however, total soil C and N storage have yet to be affected by N input. Added N plots initially had significantly lower C and N storage than control plots; however, rates of aboveground N and C storage became significantly higher added N plots after 4-5 years of exposure. N losses from leaching continue to be significantly higher in added N plots even with an increase in aboveground C and N storage. The impact of N enrichment on ecosystem C and N storage varied depending on the stage of succession, but the eventual N-induced increase in NPP has implications for fuel buildup and future fire intensity. While N enrichment acted to increase aboveground C and N storage, plots exposed to high N inputs lost substantially more N from leaching than control plots. These results indicate that post-fire chaparral shrublands tend to be "leaky" even though they are not yet "N-saturated." Recovering stands in high-N deposition areas will likely be large sources of N to groundwater and/or streams regardless of whether NPP is stimulated by N input.
Distribution of di(2-ethylhexyl) phthalate and products in blood and blood components.
Rock, G; Labow, R S; Tocchi, M
1986-01-01
In order to impart flexibility, plastic medical devices incorporate liquid plasticizers into their structure. Data from several laboratories, including ours, have shown that these compounds leach from blood bags and tubing during collection of blood, storage of various blood components and during kidney dialysis and cell and plasma apheresis procedures. After the plasticizer di(2-ethylhexyl) phthalate leaches from poly(vinyl chloride) blood packs, it is converted by a plasma enzyme to a more toxic metabolite, mono(2-ethylhexyl) phthalate. Blood fractionation products from outdated plasma contain mono(2-ethylhexyl) phthalate, the highest level being found in normal serum albumin. Recently, we have reported that di(2-ethylhexyl) phthalate actually binds to the red blood cell membrane and reduces its osmotic fragility. Current methods of red cells storage, which permit utilization up to 35 days after collection, are not possible without this membrane stabilization. Platelets are now stored for 5 days in the Fenwal PL 732 polyolefin bag. Although stated to be essentially free of liquid plasticizers, a significant level of leaching from this bag into the extracts of stored platelet concentrates was observed. PMID:3709456
Capacity estimation of soil organic carbon pools in the intertidal zone of the Bohai Bay
NASA Astrophysics Data System (ADS)
Tian-Yu, Mao; Ting-Ting, Shi; Ya-Juan, Li
2018-03-01
Based on the data obtained from the field survey in the intertidal zone of the Binhai New Area of Tianjin Bay in October 2014, the distribution characteristics of soil organic carbon pool in intertidal zone were studied. The results showed that the highest organic carbon content of soil is 22.913g/kg; the average is 16.304g/kg. The soil organic carbon pool in the intertidal zone is in the 6.58-30.40kg/m3, almost close the level of forest soil in the Binhai New Area. Moreover, close to the surrounding wetland such as Yellow River Estuary or Liaohe River Estuary. In conclusion, the soil carbon storage of the beach tidal flats is higher in the coastal zone, and the carbon storage will be significantly reduced after artificial backfilling.
NASA Astrophysics Data System (ADS)
Gabriel, C. E.; Kellman, L. M.; Ziegler, S.
2016-12-01
Mineral soil organic matter (SOM) is associated with a suite of secondary minerals that can confer stability, resulting in the potential for long-term storage of carbon (C). Not all interactions impart the same level of stability, however; evidence is suggesting that SOM in certain mineral phases is dynamic and vulnerable to soil disturbance, such as forest harvesting. The objective of this research was to characterize SOM-mineral interactions in horizons of harvested soils of contrasting stand age. Sequential selective dissolutions representing increasingly stable SOM pools from soluble minerals (deionized water (DI)), non-crystalline (Na-pyrophosphate), poorly-crystalline minerals (HCl hydroxylamine), to crystalline secondary minerals (Na-dithionite HCl)) were carried out for Ae, Bf and BC horizons sampled from a young and mature forest site (35 and 110 years post-harvest) in Mooseland, Nova Scotia, Canada. Selective dissolution extracts were analyzed for dissolved organic carbon (DOC), its δ13C, Fe and Al. Initial isotopic analysis indicates that separate operational SOM pools were isolated: δ13C values of pyrophosphate-extracted non-crystalline (NC) phases were -27 to -28‰, similar to δ13C of bulk C and to plant-derived humic acids and fungal biomass, whereas the δ13C of DI extracts were more depleted in 13C (1-2 ‰). These SOM pools retained their isotopic signature through depth despite an enrichment in bulk SOM δ13C. NC dominated the C distribution for all horizons, followed by poorly crystalline (PC) minerals, and the C content of these two phases explained the variation in bulk C, while C in crystalline pools were similar for the two sites through depth. The mature site had twice as much C in the NC pool as the young site in the Bf horizons, supported by higher C/Fe+Al ratios, suggesting a change in loading following harvesting. Despite the destabilizing processes that occur with forest harvesting and evidence for the increased destabilization of NC and PC pools of SOM, those pools associated with crystalline OM remain stable, suggesting that the nature of mineral-SOM binding determines its stability and therefore its potential for long-term storage.
Case series of ultrasound-guided platelet-rich plasma injections for sacroiliac joint dysfunction.
Ko, Gordon D; Mindra, Sean; Lawson, Gordon E; Whitmore, Scott; Arseneau, Leigh
2017-01-01
Two-thirds of adults worldwide will experience low back pain at some point in their life. In the following case series, we present four patients with sacroiliac (SI) joint instability and severe chronic low back pain, which was refractory to other treatment modalities. We investigated the efficacy of platelet-rich plasma (PRP) injections, a novel orthobiologic therapy, for reducing SI joint pain, improving quality of life, and maintaining a clinical effect. Short-form McGill Pain Questionnaire (SFM), Numeric Rating Scale (NRS), and Oswestry Low Back Pain and Disability Index were used for evaluation of treatment at pretreatment, 12-months and 48-months after treatment. At follow-up 12-months post-treatment, pooled data from all patients reported a marked improvement in joint stability, a statistically significant reduction in pain, and improvement in quality of life. The clinical benefits of PRP were still significant at 4-years post-treatment. Platelet-rich plasma therapy exhibits clinical usefulness in both pain reduction and for functional improvement in patients with chronic SI joint pain. The improvement in joint stability and low back pain was maintained at 1- and 4-years post-treatment.
Roethig, Hans J; Koval, Tamara; Muhammad-Kah, Raheema; Jin, Yan; Mendes, Paul; Unverdorben, Martin
2010-01-01
Previous studies indicate that cigarette smokers have a 5-30% higher white blood cell counts (WBC) compared to non-smokers and higher red blood cell counts. This study was to pool hematology data from three similar studies and analyze the data for effects on WBC, its subpopulations, platelets, red blood cell count (RBC) and hematocrit in adult cigarette smokers three days after using an electrically heated cigarette smoking system (EHCSS) as a potential reduced exposure product (PREP) or no-smoking compared to smoking a conventional cigarette. Lower exposure to cigarette smoke in adult, long term smokers, by using an EHCSS or stopping smoking, leads to statistically significant decreases of up to 9% in WBC, neutrophils, lymphocytes, platelets, RBC and hematocrit within three days. Switching from CC-smoking to EHCSS-smoking or no-smoking resulted in lower WBC and vice versa within 3 days. This clinical model may be used as a screening tool to find new technologies that could provide insights on changes in inflammation resulting from the change in cigarette smoke. Copyright 2010 Elsevier Inc. All rights reserved.
Federal Register 2010, 2011, 2012, 2013, 2014
2013-02-11
... Administrative Rules Regarding the Transfer and Storage of Excess Spearmint Oil AGENCY: Agricultural Marketing... such oil to the Committee or its designees for storage, from November 1 to December 1. This rule also... storage, from November 1 to December 1. This rule also changes the date that the Committee must pool...
Daniel M. Kashian; William H. Romme; Daniel B. Tinker; Monica G. Turner; Michael G. Ryan
2013-01-01
A warming climate may increase the frequency and severity of stand-replacing wildfires, reducing carbon (C) storage in forest ecosystems. Understanding the variability of postfire C cycling on heterogeneous landscapes is critical for predicting changes in C storage with more frequent disturbance. We measured C pools and fluxes for 77 lodgepole pine (Pinus contorta...
Kim, Haegyeom; Lim, Hee-Dae; Kim, Sung-Wook; Hong, Jihyun; Seo, Dong-Hwa; Kim, Dae-chul; Jeon, Seokwoo; Park, Sungjin; Kang, Kisuk
2013-01-01
High-performance and cost-effective rechargeable batteries are key to the success of electric vehicles and large-scale energy storage systems. Extensive research has focused on the development of (i) new high-energy electrodes that can store more lithium or (ii) high-power nano-structured electrodes hybridized with carbonaceous materials. However, the current status of lithium batteries based on redox reactions of heavy transition metals still remains far below the demands required for the proposed applications. Herein, we present a novel approach using tunable functional groups on graphene nano-platelets as redox centers. The electrode can deliver high capacity of ~250 mAh g−1, power of ~20 kW kg−1 in an acceptable cathode voltage range, and provide excellent cyclability up to thousands of repeated charge/discharge cycles. The simple, mass-scalable synthetic route for the functionalized graphene nano-platelets proposed in this work suggests that the graphene cathode can be a promising new class of electrode. PMID:23514953
Swarm Intelligence-Enhanced Detection of Non-Small-Cell Lung Cancer Using Tumor-Educated Platelets.
Best, Myron G; Sol, Nik; In 't Veld, Sjors G J G; Vancura, Adrienne; Muller, Mirte; Niemeijer, Anna-Larissa N; Fejes, Aniko V; Tjon Kon Fat, Lee-Ann; Huis In 't Veld, Anna E; Leurs, Cyra; Le Large, Tessa Y; Meijer, Laura L; Kooi, Irsan E; Rustenburg, François; Schellen, Pepijn; Verschueren, Heleen; Post, Edward; Wedekind, Laurine E; Bracht, Jillian; Esenkbrink, Michelle; Wils, Leon; Favaro, Francesca; Schoonhoven, Jilian D; Tannous, Jihane; Meijers-Heijboer, Hanne; Kazemier, Geert; Giovannetti, Elisa; Reijneveld, Jaap C; Idema, Sander; Killestein, Joep; Heger, Michal; de Jager, Saskia C; Urbanus, Rolf T; Hoefer, Imo E; Pasterkamp, Gerard; Mannhalter, Christine; Gomez-Arroyo, Jose; Bogaard, Harm-Jan; Noske, David P; Vandertop, W Peter; van den Broek, Daan; Ylstra, Bauke; Nilsson, R Jonas A; Wesseling, Pieter; Karachaliou, Niki; Rosell, Rafael; Lee-Lewandrowski, Elizabeth; Lewandrowski, Kent B; Tannous, Bakhos A; de Langen, Adrianus J; Smit, Egbert F; van den Heuvel, Michel M; Wurdinger, Thomas
2017-08-14
Blood-based liquid biopsies, including tumor-educated blood platelets (TEPs), have emerged as promising biomarker sources for non-invasive detection of cancer. Here we demonstrate that particle-swarm optimization (PSO)-enhanced algorithms enable efficient selection of RNA biomarker panels from platelet RNA-sequencing libraries (n = 779). This resulted in accurate TEP-based detection of early- and late-stage non-small-cell lung cancer (n = 518 late-stage validation cohort, accuracy, 88%; AUC, 0.94; 95% CI, 0.92-0.96; p < 0.001; n = 106 early-stage validation cohort, accuracy, 81%; AUC, 0.89; 95% CI, 0.83-0.95; p < 0.001), independent of age of the individuals, smoking habits, whole-blood storage time, and various inflammatory conditions. PSO enabled selection of gene panels to diagnose cancer from TEPs, suggesting that swarm intelligence may also benefit the optimization of diagnostics readout of other liquid biopsy biosources. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.
Kim, Jinseon; Kwon, Sanghyuk; Cho, Dae-Hyun; Kang, Byunggil; Kwon, Hyukjoon; Kim, Youngchan; Park, Sung O; Jung, Gwan Yeong; Shin, Eunhye; Kim, Wan-Gu; Lee, Hyungdong; Ryu, Gyeong Hee; Choi, Minseok; Kim, Tae Hyeong; Oh, Junghoon; Park, Sungjin; Kwak, Sang Kyu; Yoon, Suk Wang; Byun, Doyoung; Lee, Zonghoon; Lee, Changgu
2015-09-15
The high-volume synthesis of two-dimensional (2D) materials in the form of platelets is desirable for various applications. While water is considered an ideal dispersion medium, due to its abundance and low cost, the hydrophobicity of platelet surfaces has prohibited its widespread use. Here we exfoliate 2D materials directly in pure water without using any chemicals or surfactants. In order to exfoliate and disperse the materials in water, we elevate the temperature of the sonication bath, and introduce energy via the dissipation of sonic waves. Storage stability greater than one month is achieved through the maintenance of high temperatures, and through atomic and molecular level simulations, we further discover that good solubility in water is maintained due to the presence of platelet surface charges as a result of edge functionalization or intrinsic polarity. Finally, we demonstrate inkjet printing on hard and flexible substrates as a potential application of water-dispersed 2D materials.
Anitua, Eduardo; de la Fuente, María; Riestra, Ana; Merayo-Lloves, Jesús; Muruzábal, Francisco; Orive, Gorka
2015-09-01
To analyze whether plasma rich in growth factors (PRGF) eye drops preserve their biological characteristics and activity after storage for 3 and 6 months at -20°C, at 4°C, and at room temperature for 72 hours, compared with fresh samples (t0). Blood from 6 healthy donors was harvested and centrifuged to obtain PRGF free of leukocytes. Resulting PRGF eye drops were stored for 3 and 6 months at -20°C. At each time, 2 aliquots were maintained at room temperature or at 4°C for 72 hours. Platelet-derived growth factor-AB, transforming growth factor-β1, vascular endothelial growth factor, epidermal growth factor, insulin-like growth factor-1, angiopoietin-1, and thrombospondin-1 were quantified at each time and temperature of storage. Also, the effect of PRGF eye drops on proliferation of primary human keratocytes was evaluated. All the analyzed growth factor levels remained constant at each time and storage condition. No differences were observed in the proliferative activity of keratocytes after treatment with PRGF eye drops at any studied time or temperature. Finally, there was no microbial contamination in any of the PRGF eye drops. The preservation of the PRGF eye drops at -20°C for up to 3 and 6 months does not mean reduction of the main growth factors and proteins implicated in ocular surface wound healing. Eye drop characteristics and in vitro biological activity were not affected by their usage and conservation for 72 hours at 4°C or at room temperature.
Brisbois, Elizabeth J.; Handa, Hitesh; Major, Terry C.; Bartlett, Robert H.; Meyerhoff, Mark E.
2013-01-01
Nitric oxide (NO) is known to be a potent inhibitor of platelet activation and adhesion. Healthy endothelial cells that line the inner walls of all blood vessels exhibit a NO flux of 0.5~4×10−10 mol cm−2 min−1 that helps prevent thrombosis. Materials with a NO flux that is equivalent to this level are expected to exhibit similar anti-thrombotic properties. In this study, five biomedical grade polymers doped with S-nitroso-N-acetylpenicillamine (SNAP) were investigated for their potential to control the release of NO from the SNAP within the polymers, and further control the release of SNAP itself. SNAP in the Elast-eon E2As polymer creates an inexpensive, homogeneous coating that can locally deliver NO (via thermal and photochemical reactions) as well slowly release SNAP. Furthermore, SNAP is surprisingly stable in the E2As polymer, retaining 82% of the initial SNAP after 2 months storage at 37°C. The E2As polymer containing SNAP was coated on the walls of extracorporeal circuits (ECC) and exposed to 4 h blood flow in a rabbit model of extracorporeal circulation to examine the effects on platelet count, platelet function, clot area, and fibrinogen adsorption. After 4 h, platelet count was preserved at 100±7% of baseline for the SNAP/E2As coated loops, compared to 60±6% for E2As control circuits (n=4). The SNAP/E2As coating also reduced the thrombus area when compared to the control (2.3±0.6 and 3.4±1.1 pixels/cm2, respectively). The results suggest that the new SNAP/E2As coating has potential to improve the thromboresistance of intravascular catheters, grafts, and other blood contacting medical devices, and exhibits excellent storage stability compared to previously reported NO release polymeric materials. PMID:23777908
Characterization of neutron sources from spent fuel casks. [Skyshine
DOE Office of Scientific and Technical Information (OSTI.GOV)
Parks, C.V.; Pace, J.V. III
1987-01-01
In the interim period prior to the acceptance of spent fuel for disposal by the USDOE, utilities are beginning to choose dry cask storage as an alternative to pool re-racking, transshipments, or new pool construction. In addition, the current MRS proposal calls for interim dry storage of consolidated spent fuel in concrete casks. As part of the licensing requirements for these cask storage facilities, calculations are typically necessary to determine the yearly radiation dose received at the site boundary. Unlike wet facilities, neutron skyshine can be an important contribution to the total boundary dose from a dry storage facility. Calculationmore » of the neutron skyshine is in turn heavily dependent on the source characteristics and source model selected for the analysis. This paper presents the basic source characteristics of the spent fuel stored in dry casks and discusses factors that must be considered in evaluating and modeling the radiation sources for the subsequent skyshine calculation. 4 refs., 1 tab.« less
Muhr, Jan; Messier, Christian; Delagrange, Sylvain; Trumbore, Susan; Xu, Xiaomei; Hartmann, Henrik
2016-03-01
While trees store substantial amounts of nonstructural carbon (NSC) for later use, storage regulation and mobilization of stored NSC in long-lived organisms like trees are still not well understood. At two different sites with sugar maple (Acer saccharum), we investigated ascending sap (sugar concentration, δ(13) C, Δ(14) C) as the mobilized component of stored stem NSC during early springtime. Using the bomb-spike radiocarbon approach we were able to estimate the average time elapsed since the mobilized carbon (C) was originally fixed from the atmosphere and to infer the turnover time of stem storage. Sites differed in concentration dynamics and overall δ(13) C, indicating different growing conditions. The absence of temporal trends for δ(13) C and Δ(14) C indicated sugar mobilization from a well-mixed pool with average Δ(14) C consistent with a mean turnover time (TT) of three to five years for this pool, with only minor differences between the sites. Sugar maple trees hence appear well buffered against single or even several years of negative plant C balance from environmental stress such as drought or repeated defoliation by insects. Manipulative investigations (e.g. starvation via girdling) combined with Δ(14) C measurements of this mobilized storage pool will provide further new insights into tree storage regulation and functioning. © 2015 The Authors. New Phytologist © 2015 New Phytologist Trust.
Optimization of spent fuel pool weir gate driving mechanism
NASA Astrophysics Data System (ADS)
Liu, Chao; Du, Lin; Tao, Xinlei; Wang, Shijie; Shang, Ertao; Yu, Jianjiang
2018-04-01
Spent fuel pool is crucial facility for fuel storage and nuclear safety, and the spent fuel pool weir gate is the key related equipment. In order to achieve a goal of more efficient driving force transfer, loading during the opening/closing process is analyzed and an optimized calculation method for dimensions of driving mechanism is proposed. The result of optimizing example shows that the method can be applied to weir gates' design with similar driving mechanism.
Whendee L. Silver; Jason Neff; Megan McGroddy; Ed Veldkamp; Michael Keller; Raimundo Cosme
2000-01-01
Soil texture plays a key role in belowground C storage in forest ecosystems and strongly influences nutrient availability and retention, particularly in highly weathered soils. We used field data and the Century ecosystem model to explore the role of soil texture in belowground C storage, nutrient pool sizes, and N fluxes in highly weathered soils in an Amazonian...
Carbon dioxide emitted from live stems of tropical trees is several years old.
Muhr, Jan; Angert, Alon; Negrón-Juárez, Robinson I; Muñoz, Waldemar Alegria; Kraemer, Guido; Chambers, Jeffrey Q; Trumbore, Susan E
2013-07-01
Storage carbon (C) pools are often assumed to contribute to respiration and growth when assimilation is insufficient to meet the current C demand. However, little is known of the age of stored C and the degree to which it supports respiration in general. We used bomb radiocarbon ((14)C) measurements to determine the mean age of carbon in CO2 emitted from and within stems of three tropical tree species in Peru. Carbon pools fixed >1 year previously contributed to stem CO2 efflux in all trees investigated, in both dry and wet seasons. The average age, i.e., the time elapsed since original fixation of CO2 from the atmosphere by the plant to its loss from the stem, ranged from 0 to 6 years. The average age of CO2 sampled 5-cm deep within the stems ranged from 2 to 6 years for two of the three species, while CO2 in the stem of the third tree species was fixed from 14 to >20 years previously. Given the consistency of (14)C values observed for individuals within each species, it is unlikely that decomposition is the source of the older CO2. Our results are in accordance with other studies that have demonstrated the contribution of storage reserves to the construction of stem wood and root respiration in temperate and boreal forests. We postulate the high (14)C values observed in stem CO2 efflux and stem-internal CO2 result from respiration of storage C pools within the tree. The observed age differences between emitted and stem-internal CO2 indicate an age gradient for sources of CO2 within the tree: CO2 produced in the outer region of the stem is younger, originating from more recent assimilates, whereas the CO2 found deeper within the stem is older, fueled by several-year-old C pools. The CO2 emitted at the stem-atmosphere interface represents a mixture of young and old CO2. These observations were independent of season, even during a time of severe regional drought. Therefore, we postulate that the use of storage C for respiration occurs on a regular basis challenging the assumption that storage pools serve as substrates for respiration only during times of limited assimilation.
NASA Astrophysics Data System (ADS)
Hutchinson, A.; Woodside, G.; Ralph, F. M.
2017-12-01
Stormwater represents a significant source of water used by the Orange County Water District (OCWD) to recharge the Orange County groundwater basin. Over the last 20 years, OCWD has captured and recharged an average of 50,000 acre-feet per year (afy) of stormwater. Much of this recharge is made possible by the capture of stormwater in the Prado Dam Conservation Pool. OCWD has and continues to work closely with the US Army Corps of Engineers (USACE) to manage the conservation pool and to increase the amount of water that can be temporarily impounded in the conservation pool. Currently, the Conservation Pool is allowed to rise to elevation 498 ft msl (approx. 10,000 af of storage) during the storm season and to 505 ft msl (approx. 20,000 af of storage) during the non-storm season. OCWD has been working with the USACE on a Feasibility Study to permanently allow for storage of stormwater up to elevation 505 msl year-round. Even though increasing the Conservation Pool will increase the amount of stormwater captured, the weather forecasting used to manage the conservation pool can be improved in order to minimize lost opportunities to capture water or unnecessary releases of water to the ocean. To increase the efficiency of stormwater capture, OCWD is partnering with the Center for Western Weather and Water Extremes (http://cw3e.ucsd.edu/) to study the viability of using Forecast-Informed Reservoir Operations (FIRO) at Prado Dam. FIRO represents the next generation of operating water reservoirs using the best available technology. Moreover, given the importance of atmospheric river (AR) storms on water supplies in California, FIRO represents a methodology to take advantage of our increasing understanding of AR storms which are infrequent but provide a large percentage of total precipitation.
Jawahar, P; Balasubramanian, M
2006-12-01
Glass fiber reinforced polyester composite and hybrid nanoclay-fiber reinforced composites were prepared by hand lay-up process. The mechanical behavior of these materials and the changes as a result of the incorporation of both nanosize clay and glass fibers were investigated. Composites were prepared with a glass fibre content of 25 vol%. The proportion of the nanosize clay platelets was varied from 0.5 to 2.5 vol%. Hybrid clay-fiber reinforced polyester composite posses better tensile, flexural, impact, and barrier properties. Hybrid clay-fiber reinforced polyester composites also posses better shear strength, storage modulus, and glass transition temperature. The optimum properties were found to be with the hybrid laminates containing 1.5 vol% nanosize clay.
The Protective Effect of Poloxamer-188 on Platelet Functions.
Guler, Nil; Abro, Schuharazad; Emanuele, Marty; Iqbal, Omer; Hoppensteadt, Debra; Fareed, Jawed
2017-11-01
Poloxamer-188 (MST-188) is effective in the repair/recovery of damaged cell membranes. MST-188 is a promising agent for protecting blood cell viability. The aim of the study is to test the hypothesis that MST-188 can extend the duration of platelet function. Blood samples were collected from 20 healthy volunteers. MST-188 (10 or 2 mg/mL) containing platelet-rich plasma (PRP) was prepared with 2 procedures. First, PRP prepared from MST-188 added whole blood (WB); second, MST-188 was added to PRP. These were referred to MST-188-WB preparation (WBP) and MST-188-PRP preparation (PRPP), respectively. For control, saline was used in the same manner. Agonist-induced aggregation (AIA) studies were performed at 30, 180, and 300 minutes using Platelet Aggregation Profiler (PAP-8) aggregometer (Bio/Data Corporation, Horsham, Pennsylvania) and Adenosine diphosphate (ADP), arachidonic acid, collagen, and epinephrine as agonists at final concentration of 20 µM, 500 µg/mL, 0.19 mg/mL, and 100 µM, respectively. There was a protective effect of MST-188 on ADP and collagen AIA. At 300 minutes, ADP AIA was found to be 50.2% higher than saline control in 2-mg WBP, 43% at 10-mg PRPP, and 10.4% at 2-mg PRPP. Protective effect of on collagen AIA was 65.9% in 2-mg WBP, 42.74% at 10-mg PRPP, and 11.42% at 2-mg PRPP. In comparison between 30 and 300 minutes, MST-188 showed significant protection in terms of ADP and collagen receptors and for both types of preparations (WBP and PRPP). The protective effects of MST-188 on ADP- and collagen-induced platelet aggregation may contribute to the preservation of platelet functionality upon storage in blood banks.
Deori, Kalyanjyoti; Ujjain, Sanjeev Kumar; Sharma, Raj Kishore; Deka, Sasanka
2013-11-13
Cubic spinel Co3O4 nanoparticles with spherical (0D) and hexagonal platelet (2D) morphologies were synthesized using a simple solvothermal method by tuning the reaction time. XRD and HRTEM analyses revealed pure phase with growth of Co3O4 particles along [111] and [110] directions. UV-vis studies showed two clear optical absorption peaks corresponding to two optical band gaps in the range of 400-500 nm and 700-800 nm, respectively, related to the ligand to metal charge transfer events (O(2-) → Co(2+,3+)). Under the electrochemical study in two electrode assembly system (Co3O4/KOH/Co3O4) without adding any large area support or a conductive filler, the hexagonal platelet Co3O4 particles exhibited comparatively better characteristics with high specific capacitance (476 F g(-1)), energy density 42.3 Wh kg(-1) and power density 1.56 kW kg(-1) at current density of 0.5 Ag(-1), that suited for potential applications in supercapacitors. The observed better electrochemical properties of the nanoporous Co3O4 particles is attributed to the layered platelet structural arrangement of the hexagonal platelet and the presence of exceptionally high numbers of regularly ordered pores.
Evaluation of Plasma Platelet Microparticles in Thrombotic Thrombocytopenic Purpura.
Tahmasbi, Leila; Karimi, Mehran; Kafiabadi, Sedigheh Amini; Nikougoftar, Mahin; Haghpanah, Sezaneh; Ranjbaran, Reza; Moghadam, Mohamad
2017-01-01
Platelet microparticles (PMPs) have a procoagulant activity about 50-100 times greater than active platelets due to high expression of negatively charged phospholipids on their surfaces. In this study, we evaluated microparticle immunophenotyping and also plasma PMPs level in patients with Thrombotic Thrombocytopenic Purpura (TTP) in Southern Iran. We had two study groups: 15 TTP patients and 15 healthy control group and PMPs from platelet concentrate (PC) at the 5 th day of storage. Microparticles were prepared in two steps, by low and high centrifugation followed by size confirmation via 'Dynamic Light Scattering (DLS)' Zetasizer. Immunophenotyping of PMPs was done via flow cytometry, using a FACS Calibur flow cytometer (BD, USA). PMPs counts were obtained using Partec-cyflow and Polysciences Microbeads (1 micron in diameter). Results were analyzed using FlowJo 7.6 (Treestar, USA) and Partec FlowMax software. Our results showed that the majority of microparticles in TTP patients and normal individuals were PMPs and also demonstrated that the plasma PMPs level in TTP patients was higher than the normal control group ( P -value<0.001). It seems that elevated PMPs level in TTP patients could be related to thrombotic events. Nevertheless, more studies are needed to confirm these results. © 2017 by the Association of Clinical Scientists, Inc.
Blood transfusion in trauma patients: unresolved questions.
Cushing, M; Shaz, B H
2011-03-01
Massive transfusion is an essential part of resuscitation efforts in acute trauma patients. The goal is to quickly correct trauma-induced coagulopathy and replace red blood cell (RBC) mass with the minimal number as well as the appropriate choice of blood components to minimize the possible adverse effects of transfusions. Early trauma induced coagulopathy (ETIC) is present in about 20% of patients upon hospital admission and predicts for decreased survival. The mechanism of ETIC is still being elucidated; however, most theories of ETIC's pathophysiology justify the early use of plasma. Most massive transfusion protocol (MTP) ratios deliver blood products in a ratio of 1:1:1 for RBCs:plasma:platelets, which is supported by the majority of the literature demonstrating improved patient survival with higher ratios (>1 plasma and platelet for every 2 RBCs transfused). Indeed, formula-driven MTPs allow trauma services to react quickly to ETIC and provide coagulation factors and platelets in these ratios without having to wait for the results of coagulation assays while the patient's coagulopathy worsens. New MTPs are being created which are adjusted according to an individual's coagulation laboratory values based on point-of-care laboratory tests, such as thromboelastography. When creating an MTP, product wastage due to inappropriate activation and improper product storage should be considered and closely monitored. Another area of discussion regarding transfusion in trauma includes the potential association of prolonged storage of RBCs and adverse outcomes, which has yet to be confirmed. Significant progress has been made in the transfusion management of trauma patients, but further studies are required to optimize patient care and outcomes.
Devasahayam, Raj; Georges, Pierre; Hodge, Christopher; Treloggen, Jane; Cooper, Simon; Petsoglou, Con; Sutton, Gerard; Zhu, Meidong
2016-09-01
Organ Culture corneal storage offers an extended storage time and increased donor pool and tissue assessment opportunities. In September 2011, the Lions New South Wales Eye Bank (LNSWEB) moved from hypothermic storage to Organ Culture corneal storage. This study evaluates the impact of implementation of Organ Culture on donor eye retrieval and the corneal transplant waiting list over a 3 year period in NSW, Australia. Retrospective review of the LNSWEB data from September 2011 to August 2014. Tissue collection, waiting list and tissue utilization data were recorded. The data from September 2008 to August 2011 for Optisol-GS storage was used for comparison. The annual donor and cornea collection rate increased 35 % and 44 % respectively with Organ Culture compared to Optisol-GS storage. The utilization rate of corneal tissue increased from 73.4 % with hypothermic storage to 77.2 % with Organ Culture storage. The transplant wait list decreased by 77.3 % from September 2011 to August 2014 and correlated with the increased rate of corneal transplantation (r = -0.9381, p < 0.0001). No other factors impacting the wait list changed over this period. Corneas not used from either storage method were due to unacceptable endothelial cell density/viability. The contamination rate of corneas stored in Organ Culture medium was low at 1.74 %. The Organ Culture storage method increases the corneal donor pool available to Eye banks. The practical benefits of the extended storage time and increased donor assessment opportunities have directly led to an increase in corneal utilization rate and a significant decrease in recipient wait list time.
Code of Federal Regulations, 2011 CFR
2011-07-01
... shall have the meaning given them in the Act and in this section. Capacity means the volume of liquid... or emptying means the partial or complete removal of stored liquid from a storage vessel. Storage vessels that contain liquid only as wall or bottom clingage, or in pools due to bottom irregularities, are...
Code of Federal Regulations, 2013 CFR
2013-07-01
... shall have the meaning given them in the Act and in this section. Capacity means the volume of liquid... or emptying means the partial or complete removal of stored liquid from a storage vessel. Storage vessels that contain liquid only as wall or bottom clingage, or in pools due to bottom irregularities, are...
Code of Federal Regulations, 2014 CFR
2014-07-01
... shall have the meaning given them in the Act and in this section. Capacity means the volume of liquid... or emptying means the partial or complete removal of stored liquid from a storage vessel. Storage vessels that contain liquid only as wall or bottom clingage, or in pools due to bottom irregularities, are...
Code of Federal Regulations, 2010 CFR
2010-07-01
... shall have the meaning given them in the Act and in this section. Capacity means the volume of liquid... or emptying means the partial or complete removal of stored liquid from a storage vessel. Storage vessels that contain liquid only as wall or bottom clingage, or in pools due to bottom irregularities, are...
Code of Federal Regulations, 2012 CFR
2012-07-01
... shall have the meaning given them in the Act and in this section. Capacity means the volume of liquid... or emptying means the partial or complete removal of stored liquid from a storage vessel. Storage vessels that contain liquid only as wall or bottom clingage, or in pools due to bottom irregularities, are...
Afforestation effects on soil carbon storage in the United States: a synthesis
L.E. Nave; C.W. Swanston; U. Mishra; K.J. Nadelhoffer
2013-01-01
Afforestation (tree establishment on nonforested land) is a management option for increasing terrestrial C sequestration and mitigating rising atmospheric carbon dioxide because, compared to nonforested land uses, afforestation increases C storage in aboveground pools. However, because terrestrial ecosystems typically store most of their C in soils, afforestation...
Bacterial detection of platelets: current problems and possible resolutions.
Blajchman, Morris A; Beckers, Erik A M; Dickmeiss, Ebbe; Lin, Lilly; Moore, Gillian; Muylle, Ludo
2005-10-01
The greatest transfusion-transmitted disease risk facing a transfusion recipient is that of bacterial sepsis. The prevalence of bacterial contamination in platelets and red blood cells is approximately 1 in 3,000 units transfused. The available data indicate that transfusion-associated sepsis develops after 1 in 25,000 platelet transfusions and 1 in 250,000 red blood cell transfusions. One of the most widely used strategies for decreasing bacterial sepsis risk is bacterial detection. A roundtable meeting of experts was convened during the XXVIII Annual Congress of the International Society of Blood Transfusion (Edinburgh, UK, July 2004) to provide a forum for experts to share their experiences in the routine bacterial detection of platelet products. This article summarizes the presentations, discussions, and recommendations of the panel. The data presented indicate that some of the current bacterial screening technology is useful for blocking the issuance of platelet units that contain relatively high levels of contaminating bacteria. Platelet units are usually released based on a test-negative status, which often become test-positive only upon longer storage. These data thus suggest that bacterial screening may not prevent all transfusion-transmitted bacterial infections. Two transfusion-transmitted case reports further highlighted the limitation of the routine bacterial screening of platelet products. It was felt that newer technologies, such as pathogen inactivation, may represent a more reliable process, with a higher level of safety. The panel thus recommended that the Transfusion Medicine community may need to change its thinking (paradigm) about bacterial detection, toward the possibility of the pathogen inactivation of blood products, to deal with the bacterial contamination issue. It was suggested, where permitted by regulatory agencies, that blood centers should consider adopting first-generation pathogen inactivation systems as a more effective approach to reducing the risk of transfusion-associated sepsis than some of the approaches currently available.
Gifford, Sean C; Strachan, Briony C; Xia, Hui; Vörös, Eszter; Torabian, Kian; Tomasino, Taylor A; Griffin, Gary D; Lichtiger, Benjamin; Aung, Fleur M; Shevkoplyas, Sergey S
2018-01-01
The use of centrifugation-based approaches for processing donated blood into components is routine in the industrialized world, as disparate storage conditions require the rapid separation of 'whole blood' into distinct red blood cell (RBC), platelet, and plasma products. However, the logistical complications and potential cellular damage associated with centrifugation/apheresis manufacturing of blood products are well documented. The objective of this study was to evaluate a proof-of-concept system for whole blood processing, which does not employ electromechanical parts, is easily portable, and can be operated immediately after donation with minimal human labor. In a split-unit study (n = 6), full (~500mL) units of freshly-donated whole blood were divided, with one half processed by conventional centrifugation techniques and the other with the new blood separation system. Each of these processes took 2-3 hours to complete and were performed in parallel. Blood products generated by the two approaches were compared using an extensive panel of cellular and plasma quality metrics. Comparison of nearly all RBC parameters showed no significant differences between the two approaches, although the portable system generated RBC units with a slight but statistically significant improvement in 2,3-diphosphoglyceric acid concentration (p < 0.05). More notably, several markers of platelet damage were significantly and meaningfully higher in products generated with conventional centrifugation: the increase in platelet activation (assessed via P-selectin expression in platelets before and after blood processing) was nearly 4-fold higher for platelet units produced via centrifugation, and the release of pro-inflammatory mediators (soluble CD40-ligand, thromboxane B2) was significantly higher for centrifuged platelets as well (p < 0.01). This study demonstrated that a simple, passive system for separating donated blood into components may be a viable alternative to centrifugation-particularly for applications in remote or resource-limited settings, or for patients requiring highly functional platelet product.
Peters, A L; Vervaart, M A T; van Bruggen, R; de Korte, D; Nieuwland, R; Kulik, W; Vlaar, A P J
2017-01-01
The accumulation of non-polar lipids arachidonic acid, 5-hydroxyeicosatetraenoic acid (HETE), 12-HETE and 15-HETE during storage of transfusion products may play a role in the onset of transfusion-related acute lung injury (TRALI), a syndrome of respiratory distress after transfusion. We investigated non-polar lipid accumulation in red blood cells (RBCs) stored for 42 days, plasma stored for 7 days at either 4 or 20°C and platelet (PLT) transfusion products stored for 7 days. Furthermore, we investigated whether transfusion of RBCs with increased levels of non-polar lipids induces TRALI in a 'two-hit' human volunteer model. All products were produced following Dutch Blood Bank protocols and are according to European standards. Non-polar lipids were measured with high-performance liquid chromotography followed by mass spectrometry. All non-polar lipids increased in RBCs after 21 days of storage compared to baseline. The non-polar lipid concentration in plasma increased significantly, and the increase was even more pronounced in products stored at 20°C. In platelets, baseline levels of 5-HETE and 15-HETE were higher than in RBCs or plasma. However, the non-polar lipids did not change significantly during storage of PLT products. Infusion of RBCs with increased levels of non-polar lipids did not induce TRALI in LPS-primed human volunteers. We conclude that non-polar lipids accumulate in RBC and plasma transfusion products and that accumulation is temperature dependent. Accumulation of non-polar lipids does not appear to explain the onset of TRALI (Dutch Trial Register - NTR4455). © 2016 International Society of Blood Transfusion.
Seghatchian, Jerard
2005-06-16
The Publisher regrets that this article was an accidental duplication of an article that has already been published in Transfus Apher Sci, 34 (1) 125 - 130, doi:10.1016/j.transci.2005.09.002. The duplicate article has therefore been withdrawn.
In vitro platelet quality in storage containers used for pediatric transfusions.
Weiss, Sandra; Scammell, Kenneth; Levin, Elena; Culibrk, Brankica; Zolfaghari, Sima; Gyöngyössy-Issa, Maria I C; Acker, Jason P
2012-08-01
The in vitro quality of small-volume platelet (PLT) aliquots for pediatric transfusions was assessed to determine the best practice approach. Small volumes (50 mL) of single apheresis PLT components (APCs), collected on either CaridianBCT Trima or Haemonetics MCS+ instruments, were aliquoted on Days 2, 3, 4, and 5 postcollection into Fenwal PL1240 or 4R2014 bags or 60-mL polypropylene syringes. Samples were tested for in vitro quality at their recommended expiry times (4 hr for 4R2014 bags and syringes or Day 5 for PL1240 bags). Assays included pH, CD62P expression, and metabolic measures. CD62P expression increased throughout storage in all containers. Among the small-volume containers, pH, pCO(2) , lactate, and bicarbonate varied considerably. Regardless of the day of aliquoting, pCO(2) was significantly higher and pO(2) was significantly lower in gas-impermeable syringes than other containers. No bacterial growth was detected in any sample. The quality of APCs aliquoted into small-volume containers meets regulatory requirements and is generally equivalent to that of full-volume APCs at expiry. © 2012 American Association of Blood Banks.
Sono-Koree, N K; Crist, R A; Frank, E L; Rodgers, G M; Smock, K J
2016-02-01
The serotonin release assay (SRA) is considered the gold standard laboratory test for heparin-induced thrombocytopenia (HIT). The historic SRA method uses platelets loaded with radiolabeled serotonin to evaluate platelet activation by HIT immune complexes. However, a nonradioactive method is desirable. We report the performance characteristics of a high-performance liquid chromatography (HPLC) SRA method. We validated the performance characteristics of an HPLC-SRA method, including correlation with a reference laboratory using the radioactive method. Serotonin released from reagent platelets was quantified by HPLC using fluorescent detection. Results were expressed as % release and classified as positive, negative, or indeterminate based on previously published cutoffs. Serum samples from 250 subjects with suspected HIT were tested in the HPLC-SRA and with the radioactive method. Concordant classifications were observed in 230 samples (92%). Sera from 41 healthy individuals tested negative. Between-run imprecision studies showed standard deviation of <6 (% release) for positive, weak positive, and negative serum pools. Stability studies demonstrated stability after two freeze-thaw cycles or up to a week of refrigeration. The HPLC-SRA has robust performance characteristics, equivalent to the historic radioactive method, but avoids the complexities of working with radioactivity. © 2015 John Wiley & Sons Ltd.
Selmants, Paul C; Litton, Creighton M; Giardina, Christian P; Asner, Gregory P
2014-09-01
Theory and experiment agree that climate warming will increase carbon fluxes between terrestrial ecosystems and the atmosphere. The effect of this increased exchange on terrestrial carbon storage is less predictable, with important implications for potential feedbacks to the climate system. We quantified how increased mean annual temperature (MAT) affects ecosystem carbon storage in above- and belowground live biomass and detritus across a well-constrained 5.2 °C MAT gradient in tropical montane wet forests on the Island of Hawaii. This gradient does not systematically vary in biotic or abiotic factors other than MAT (i.e. dominant vegetation, substrate type and age, soil water balance, and disturbance history), allowing us to isolate the impact of MAT on ecosystem carbon storage. Live biomass carbon did not vary predictably as a function of MAT, while detrital carbon declined by ~14 Mg of carbon ha(-1) for each 1 °C rise in temperature - a trend driven entirely by coarse woody debris and litter. The largest detrital pool, soil organic carbon, was the most stable with MAT and averaged 48% of total ecosystem carbon across the MAT gradient. Total ecosystem carbon did not vary significantly with MAT, and the distribution of ecosystem carbon between live biomass and detritus remained relatively constant across the MAT gradient at ~44% and ~56%, respectively. These findings suggest that in the absence of alterations to precipitation or disturbance regimes, the size and distribution of carbon pools in tropical montane wet forests will be less sensitive to rising MAT than predicted by ecosystem models. This article also provides needed detail on how individual carbon pools and ecosystem-level carbon storage will respond to future warming. © 2014 John Wiley & Sons Ltd.
Human Platelet Lysate as a Replacement for Fetal Bovine Serum in Limbal Stem Cell Therapy.
Suri, Kunal; Gong, Hwee K; Yuan, Ching; Kaufman, Stephen C
2016-10-01
To evaluate the use of human platelet lysate (HPL) as an alternative supplement for limbal explant culture. Culture media were prepared using either 10% pooled HPL (PHPL), single donor HPL, or fetal bovine serum (FBS). Limbal tissues, obtained from the Minnesota Lions Eye Bank, were cultured in each medium on plastic plates or on denuded amniotic membrane (AM). Immunofluorescence staining was performed for ABCG2, tumor protein p63α, and cytokeratin 3 (K3). Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) was used to evaluate the expression of ABCG2 and p63. Limbal explants grown in each medium were labeled with bromodeoxyuridine (BrdU) to assess the proliferative capacity in each medium. Concentration of growth factors including epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), transforming growth factor-β (TGF-β), and platelet derived growth factor (PDGF) in HPL and PHPL was compared to that in human serum (HS). Immunofluorescence staining on AM showed prominent expression of ABCG2, p63α but sparse expression of K3 in HPL and PHPL supplemented medium. Real time-PCR showed 1.7 fold higher expression of ABCG2 in PHPL supplemented medium (p = 0.03), and similar expression of p63 in HPL and PHPL supplemented medium compared to FBS medium. The proliferation assay showed that LSCs retained their proliferative potential in HPL supplemented medium. Higher concentration of growth factors were found in HPL, compared to HS. Human platelet lysate has higher concentration of grown factors and is effective in maintaining growth and stem cell phenotype of corneal limbal explant cultures.
Carbon storage by urban soils in the United States
Richard V. Pouyat; Ian D. Yesilonis; David J. Nowak
2006-01-01
We used data available from the literature and measurements from Baltimore, Maryland to (i) assess inter-city variability of soil oganic carbon (SOC) pools (1-m depth) of six cities (Atlanta, Baltimore, Boston, Chicago, Oakland, and Syracuse); (ii) calculate the net effect of urban land-use conversion on SOC pools for the same cities; (iii) use the National Land Cover...
Fine bed material in pools of natural gravel bed channels
Thomas E. Lisle; Sue Hilton
1999-01-01
Abstract - Natural gravel bed channels commonly contain a fine mode of sand and fine gravel that fills voids of the bed framework of coarser gravel. If the supply of fine bed material exceeds the storage capacity of framework voids, excess fine material forms surficial patches, which can be voluminous in pools during low flow. Data collected in 34 natural channels in...
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. Copyright© by Royan Institute. All rights reserved.
What do we know about the role and regulation of stored non-structural carbon compounds in trees?
NASA Astrophysics Data System (ADS)
Sala, A.; Martinez-Vilalta, J.; Lloret, F.
2012-12-01
Despite the critical role of forests on the global C cycle and recent increases in drought-induced forest mortality, remarkable knowledge gaps exist to accurately predict tree growth and survival under climate change. In particular, storage of non-structural carbon compounds (NSCC) is thought to be critical for tree survival under drought but its regulation and function is the least understood of the tree's C budget components. Our current understanding of the role and regulation of stored NSCC relies on several assumptions. First, stored NSCC is generally assumed to be a passive buffer between source and sink demand for growth and respiration and, therefore, is an integrator of the tree C balance. Second, most process-based models commonly assume that C availability drives growth and ignore storage and environmental regulation of sink activity. Third, trees under C deficits are assumed to rely on stored C until normal conditions are restored or reserves are exhausted, whichever comes first. Implicit is this is that stored NSCC increases survival under drought, and that access to stored NSCC is unlimited. For the most part, these assumptions have not been experimentally tested, and increasing evidence suggests that some of them are not necessarily correct. Here we assess the validity of some of the assumptions above from a review of the published data. Several studies so far are consistent with the notion that stored NSCC serve as a passive buffer between C assimilation and C demand for growth and respiration. In contrast, other studies indicate that C may be partitioned to storage at the expense of growth. In any case, unequivocal evidence of whether and when C is or is not partitioned to storage at the expense of growth in woody plants is lacking, leaving a critical void in our knowledge. Many studies in woody plants indicate that growth is more sensitive to water availability than photosynthesis, and that NSCC accumulate as a result. This indicates that growth is not solely driven by C assimilation as most process-based models currently assume, and that sink activity is directly affected by environmental conditions. These results also suggest that fluctuations in NSCC storage also arise as a passive response to imbalances between C assimilation and growth. Differences in relative sensitivities of growth and photosynthesis to N availability are much less clear than those to water availability. Data also suggests that woody plants rarely fully deplete their pool of stored NSCC, tentatively suggesting that they regulate their storage pool to maintain certain minimums that are well above those needed to maintain cell turgor. Most often, studies reporting nearly exhausted storage pools as a consequence of drought also report imminent death of trees. This suggests that most of the stored NSCC pool remains available for use, particularly under extreme conditions. Whether the turning point for death is when C availability is insufficient to sustain cell metabolism or when stored NSCC pools are depleted below minimum thresholds is unclear. In the latter case, the point of no recovery could occur well before their NSCC pools are exhausted, although visual death symptoms may lag behind and occur at the point of NSCC exhaustion.
Kapala, J.; Copes, D.; Sproston, A.; Patel, J.; Jang, D.; Petrich, A.; Mahony, J.; Biers, K.; Chernesky, M.
2000-01-01
Specimen pooling to achieve efficiency when testing urine specimens for Chlamydia trachomatis nucleic acids has been suggested. We pooled endocervical swabs from 1,288 women and also tested individual swabs by ligase chain reaction (LCR). Out of 53 positive specimens, pools of 4 or 8 specimens missed two positives, providing 96.2% accuracy compared to individual test results. Dilution and positive-control spiking experiments showed that negative specimens with inhibitors of LCR in the pool reduced the signal. Conversely, two extra positives, detected only through pooling, were negative by individual testing but became positive after storage, suggesting that fresh positive specimens with labile inhibitors may be positive in a pool because of dilution of inhibitors. For this population of women with a 4% prevalence of C. trachomatis infection, substantial savings in cost of reagents (55 to 63%) and technologist time (50 to 63%) made pooling strategies a desirable alternative to individual testing. PMID:10878029
Mitchell, Thomas A; Herzig, Maryanne C; Fedyk, Chriselda G; Salhanick, Marc A; Henderson, Aaron T; Parida, Bijaya K; Prat, Nicolas J; Dent, Daniel L; Schwacha, Martin G; Cap, Andrew P
2017-06-01
Autotransfusion of shed blood from traumatic hemothorax is an attractive option for resuscitation of trauma patients in austere environments. However, previous analyses revealed that shed hemothorax (HX) blood is defibrinated, thrombocytopenic, and contains elevated levels of D-dimer. Mixing studies with normal pooled plasma demonstrated hypercoagulability, evoking concern for potentiation of acute traumatic coagulopathy. We hypothesized that induction of coagulopathic changes by shed HX blood may be due to increases in cellular microparticles (MP) and that these may also affect recipient platelet function. Shed HX blood was obtained from 17 adult trauma patients under an Institutional Review Board approved prospective observational protocol. Blood samples were collected every hour up to 4 h after thoracostomy tube placement. The corresponding plasma was isolated and frozen for analysis. The effects of shed HX frozen plasma (HFP) and isolated HX microparticles (HMP) on coagulation and platelet function were assessed through mixing studies with platelet-rich plasma at various dilutions followed by analysis with thromboelastometry (ROTEM), platelet aggregometry (Multiplate), enzyme-linked immunosorbent assays, and flow cytometry. Furthermore, HFP was assessed for von Willebrand factor antigen levels and multimer content, and plasma-free hemoglobin. ROTEM analysis demonstrated that diluted HFP and isolated HMP samples decreased clotting time, clotting formation time, and increased α angle, irrespective of sample concentrations, when compared with diluted control plasma. Isolated HMP inhibited platelet aggregation in response to adenosine diphosphate, arachidonic acid, and collagen. HFP contained elevated levels of fibrin-degradation products and tissue factor compared with control fresh frozen plasma samples. MP concentrations in HFP were significantly increased and enriched in events positive for phosphatidylserine, tissue factor, CD235, CD45, CD41a, and CD14. von Willebrand factor (vWF) multimer analysis revealed significant loss of high molecular weight multimers in HFP samples. Plasma-free hemoglobin levels were 8-fold higher in HFP compared with fresh frozen plasma. HFP induces plasma hypercoagulability that is likely related to increased tissue factor and phosphatidylserine expression originating from cell-derived MP. In contrast, platelet dysfunction is induced by HMP, potentially aggravated by depletion of high molecular weight multimers of vWF. Thus, autologous transfusion of shed traumatic hemothorax blood may induce a range of undesirable effects in patients with acute traumatic coagulopathy.
Modeling evaporation from spent nuclear fuel storage pools: A diffusion approach
NASA Astrophysics Data System (ADS)
Hugo, Bruce Robert
Accurate prediction of evaporative losses from light water reactor nuclear power plant (NPP) spent fuel storage pools (SFPs) is important for activities ranging from sizing of water makeup systems during NPP design to predicting the time available to supply emergency makeup water following severe accidents. Existing correlations for predicting evaporation from water surfaces are only optimized for conditions typical of swimming pools. This new approach modeling evaporation as a diffusion process has yielded an evaporation rate model that provided a better fit of published high temperature evaporation data and measurements from two SFPs than other published evaporation correlations. Insights from treating evaporation as a diffusion process include correcting for the effects of air flow and solutes on evaporation rate. An accurate modeling of the effects of air flow on evaporation rate is required to explain the observed temperature data from the Fukushima Daiichi Unit 4 SFP during the 2011 loss of cooling event; the diffusion model of evaporation provides a significantly better fit to this data than existing evaporation models.
Vollmer, T; Engemann, J; Kleesiek, K; Dreier, J
2011-06-01
Bacterial contamination is currently the major infectious hazard of platelet transfusion in developed countries. It has been demonstrated that a significant transfusion risk remains, in particular with older platelet concentrates (PCs). In 2009, the shelf life of PCs was therefore reduced in Germany to 4 days after the day of production according to Vote 38. The aim of the present study was the application and implementation of a recently developed flow cytometry-based rapid screening method (BactiFlow) for bacterial contamination at the end of PC shelf life as a routine in-process control. A total of 472 apheresis-derived PCs were tested using the BactiFlow flow cytometric assay to detect and count bacteria based on esterase activity in viable bacterial cells, while the BacT/Alert automated culture system served as the reference method. The automation potential of the flow cytometric assay was analysed by applying the semi-automated BactiFlow ALS system. An algorithm was developed for use in routine blood bank operations to extend the storage period of PCs. Two of the 472 apheresis PCs tested were positive in culture and identified as Propionibacterium species. One PC was positive for Staphylococcus aureus by both methods. All remaining specimens were tested negative by both methods. Our study demonstrates that routine bacterial testing of PCs was successfully implemented and the established algorithm proved efficient. The BactiFlow flow cytometric assay is the first rapid screening method which is suitable for a routine application combined with a high sensitivity. © 2011 The Authors. Transfusion Medicine © 2011 British Blood Transfusion Society.
USDA-ARS?s Scientific Manuscript database
The relationship between soil organic carbon (SOC) content and quality of SOC as impacted by land management is not well understood and may influence long-term storage of SOC. To better understand the potential for SOC storage in specific aggregate pools (i.e. physically protected intra-aggregate C)...
Claire M. Ruffing; Kathleen A. Dwire; Melinda D. Daniels
2016-01-01
Streams and their accompanying riparian environments are intrinsic components of terrestrial carbon cycling. However, they have been understudied in terms of the magnitude of their storage components and the role of disturbance in determining carbon storage capacity. This study presents partial carbon budgets for stream-riparian corridors along six study...
Identifying the Dynamic Catchment Storage That Does Not Drive Runoff
NASA Astrophysics Data System (ADS)
Dralle, D.; Hahm, W. J.; Rempe, D.; Karst, N.; Thompson, S. E.; Dietrich, W. E.
2017-12-01
The central importance of subsurface water storage in hydrology has resulted in numerous attempts to develop hydrograph and mass balance based techniques to quantify catchment storage state or capacity. In spite of these efforts, relatively few studies have linked catchment scale storage metrics to Critical Zone (CZ) structure and the status of water in hillslopes. Elucidating these relationships would increase the interpretability of catchment storage metrics, and aid the development of hydrologic models. Here, we propose that catchment storage consists of a dynamic component that varies on seasonal timescales, and a static component with negligible time variation. Discharge is assumed to be explicitly sensitive to changes in some fraction of the dynamic storage, while the remaining dynamic storage varies without directly influencing flow. We use a coupled mass balance and storage-discharge function approach to partition dynamic storage between these driving and non-driving storage pools, and compare inferences with direct observations of saturated and unsaturated dynamic water storages at two field sites in Northern California. We find that most dynamic catchment water storage does not drive streamflow in both sites, even during the wettest times of year. Moreover, the physical character of non-driving dynamic storage depends strongly on catchment CZ structure. At a site with a deep profile of weathered rock, the dynamic storage that drives streamflow occurs as a seasonally perched groundwater table atop fresh bedrock, and that which does not drive streamflow resides as seasonally dynamic unsaturated water in shallow soils and deep, weathered rock. At a second site with a relatively thin weathered zone, water tables rapidly rise to intersect the ground surface with the first rains of the wet season, yet only a small fraction of this dynamic saturated zone storage drives streamflow. Our findings emphasize how CZ structure governs the overlap in time and space of three pools of subsurface water: (i) seasonally dynamic vs. static; (ii) unsaturated vs. saturated, and (iii) storage whose magnitude directly influences runoff vs. that which does not. These results highlight the importance of hillslope monitoring for physically interpreting methods of runoff-based hydrologic analysis.
Glycomic Analysis of Prostate Cancer
2012-07-01
allowed measurements of N-glycans and the Clinical Molecular Epidemiology Shared Resources which provided services for biological sample storage and...select N-glycans for the detection of prostate cancer. Aim3. Perform an exploratory study of N-glycans in urine of the participants and correlation of...cases. We have designed a pooled-unpooled study where initial discovery is conducted in smaller number of pooled samples followed by analysis of
2012-01-01
Background Global forests capture and store significant amounts of CO2 through photosynthesis. When carbon is removed from forests through harvest, a portion of the harvested carbon is stored in wood products, often for many decades. The United States Forest Service (USFS) and other agencies are interested in accurately accounting for carbon flux associated with harvested wood products (HWP) to meet greenhouse gas monitoring commitments and climate change adaptation and mitigation objectives. This paper uses the Intergovernmental Panel on Climate Change (IPCC) production accounting approach and the California Forest Project Protocol (CFPP) to estimate HWP carbon storage from 1906 to 2010 for the USFS Northern Region, which includes forests in northern Idaho, Montana, South Dakota, and eastern Washington. Results Based on the IPCC approach, carbon stocks in the HWP pool were increasing at one million megagrams of carbon (MgC) per year in the mid 1960s, with peak cumulative storage of 28 million MgC occurring in 1995. Net positive flux into the HWP pool over this period is primarily attributable to high harvest levels in the mid twentieth century. Harvest levels declined after 1970, resulting in less carbon entering the HWP pool. Since 1995, emissions from HWP at solid waste disposal sites have exceeded additions from harvesting, resulting in a decline in the total amount of carbon stored in the HWP pool. The CFPP approach shows a similar trend, with 100-year average carbon storage for each annual Northern Region harvest peaking in 1969 at 937,900 MgC, and fluctuating between 84,000 and 150,000 MgC over the last decade. Conclusions The Northern Region HWP pool is now in a period of negative net annual stock change because the decay of products harvested between 1906 and 2010 exceeds additions of carbon to the HWP pool through harvest. However, total forest carbon includes both HWP and ecosystem carbon, which may have increased over the study period. Though our emphasis is on the Northern Region, we provide a framework by which the IPCC and CFPP methods can be applied broadly at sub-national scales to other regions, land management units, or firms. PMID:22244260
Acute Fluoxetine Treatment Induces Slow Rolling of Leukocytes on Endothelium in Mice
Herr, Nadine; Mauler, Maximilian; Witsch, Thilo; Stallmann, Daniela; Schmitt, Stefanie; Mezger, Julius; Bode, Christoph; Duerschmied, Daniel
2014-01-01
Objective Activated platelets release serotonin at sites of inflammation where it acts as inflammatory mediator and enhances recruitment of neutrophils. Chronic treatment with selective serotonin reuptake inhibitors (SSRI) depletes the serotonin storage pool in platelets, leading to reduced leukocyte recruitment in murine experiments. Here, we examined the direct and acute effects of SSRI on leukocyte recruitment in murine peritonitis. Methods C57Bl/6 and Tph1−/− (Tryptophan hydroxylase1) mice underwent acute treatment with the SSRI fluoxetine or vehicle. Serotonin concentrations were measured by ELISA. Leukocyte rolling and adhesion on endothelium was analyzed by intravital microscopy in mesentery venules with and without lipopolysaccharide challenge. Leukocyte extravasation in sterile peritonitis was measured by flow cytometry of abdominal lavage fluid. Results Plasma serotonin levels were elevated 2 hours after fluoxetine treatment (0.70±0.1 µg/ml versus 0.27±0.1, p = 0.03, n = 14), while serum serotonin did not change. Without further stimulation, acute fluoxetine treatment increased the number of rolling leukocytes (63±8 versus 165±17/0.04 mm2min−1) and decreased their velocity (61±6 versus 28±1 µm/s, both p<0.0001, n = 10). In Tph1−/− mice leukocyte rolling was not significantly influenced by acute fluoxetine treatment. Stimulation with lipopolysaccharide decreased rolling velocity and induced leukocyte adhesion, which was enhanced after fluoxetine pretreatment (27±3 versus 36±2/0.04 mm2, p = 0.008, n = 10). Leukocyte extravasation in sterile peritonitis, however, was not affected by acute fluoxetine treatment. Conclusions Acute fluoxetine treatment increased plasma serotonin concentrations and promoted leukocyte-endothelial interactions in-vivo, suggesting that serotonin is a promoter of acute inflammation. E-selectin was upregulated on endothelial cells in the presence of serotonin, possibly explaining the observed increase in leukocyte-endothelial interactions. However transmigration of neutrophils in sterile peritonitis was not affected by higher serotonin concentrations, indicating that the effect of fluoxetine was restricted to early steps in the leukocyte recruitment. Whether SSRI use in humans alters leukocyte recruitment remains to be investigated. PMID:24520366
Kobayashi, J; Yanagisawa, R; Ono, T; Tatsuzawa, Y; Tokutake, Y; Kubota, N; Hidaka, E; Sakashita, K; Kojima, S; Shimodaira, S; Nakamura, T
2018-02-01
Adverse reactions to platelet transfusions are a problem. Children with primary haematological and malignant diseases may experience allergic transfusion reactions (ATRs) to platelet concentrates (PCs), which can be prevented by giving washed PCs. A new platelet additive solution, using bicarbonated Ringer's solution and acid-citrate-dextrose formula A (BRS-A), may be better for platelet washing and storage, but clinical data are scarce. A retrospective cohort study for consecutive cases was performed between 2013 and 2017. For 24 months, we transfused washed PCs containing BRS-A to children with primary haematological and malignant diseases and previous adverse reactions. Patients transfused with conventional PCs (containing residual plasma) were assigned as controls, and results were compared in terms of frequency of ATRs, corrected count increment (CCI) and occurrence of bleeding. We also studied children transfused with PCs washed by a different system as historical controls. Thirty-two patients received 377 conventional PC transfusions. ATRs occurred in 12 (37·5%) patients from transfused with 18 (4·8%) bags. Thirteen patients, who experienced reactions to regular PCs in plasma, then received 119 transfusion bags of washed PCs containing BRS-A, and none had ATRs to washed PCs containing BRS-A. Before study period, six patients transfused 137 classical washed PCs with different platelet additive solution, under same indication, ATRs occurred in one (16·7%) patient from transfused with one (0·7%) bags. CCIs (24 h) in were lower with classical washed PCs (1·26 ± 0·54) compared to regular PCs in plasma (2·07 ± 0·76) (P < 0·001), but there was no difference between washed PCs containing BRS-A (2·14 ± 0·77) and regular PCs (2·21 ± 0·79) (P = 0·769), and we saw no post-transfusion bleeding. Washed PCs containing BRS-A appear to prevent ATRs without loss of transfusion efficacy in children with primary haematological and malignant diseases. Their efficacy should be further evaluated through larger prospective clinical trials. © 2017 International Society of Blood Transfusion.
Innate immune responses of equine monocytes cultured in equine platelet lysate.
Naskou, Maria C; Norton, Natalie A; Copland, Ian B; Galipeau, Jacques; Peroni, John F
2018-01-01
Platelet lysate (PL) has been extensively used for the laboratory expansion of human mesenchymal stem cells (MSC) in order to avoid fetal bovine serum (FBS) which has been associated with immune-mediated host reactions and transmission of bovine-origin microbial contaminants. Before suggesting the routine use of PL for MSC culture, we wanted to further investigate whether PL alone might trigger inflammatory responses when exposed to reactive white blood cells such as monocytes. Our objectives were to evaluate the inflammatory profile of equine monocytes cultured with equine PL (ePL) and to determine if ePL can modulate the expression of inflammatory cytokines in lipopolysaccharide (LPS)-stimulated monocytes. In a first experiment, equine monocytes were isolated and incubated with donor horse serum (DHS), FBS, six individual donors ePL or pooled ePL from all horses. In a second experiment, monocytes were stimulated with E. coli LPS in the presence of 1, 5 or 10% DHS and/or pooled ePL. After 6h of incubation, cell culture supernatants were assayed via ELISA for production of the pro-inflammatory cytokines tumor necrosis factor alpha (TNF-α) and Interleukin 1β (IL-1β) as well as for the anti-inflammatory Interleukin 10 (IL-10). Equine monocytes incubated with pooled ePL produced significantly less TNF-α and significantly more IL-10 than monocytes incubated in FBS. A statistically significant difference was not identified for the production of IL-1β. The second experiment showed that pooled ePL added to LPS-stimulated equine monocytes resulted in a significant reduction in TNF-α and IL-1β production. IL-10 production was not significantly upregulated by the addition of ePL to LPS-stimulated monocytes. Finally, the addition of ePL to LPS-stimulated monocytes in the presence of various concentrations of DHS resulted to statistically significant decrease of TNF-α and IL-1β compared to the control groups. This is the first study to demonstrate that ePL suppresses the release of pro-inflammatory cytokines from stimulated equine monocytes. These results encourage further exploration of PL as a homologous media substitute for FBS but also opens the possibility of investigating its use as means to suppress cell-mediated inflammation. Published by Elsevier B.V.
Girona-Llobera, Enrique; Jimenez-Marco, Teresa; Galmes-Trueba, Ana; Muncunill, Josep; Serret, Carmen; Serra, Neus; Sedeño, Matilde
2014-01-01
Pathogen inactivation (PI) technology for blood components enhances blood safety by inactivating viruses, bacteria, parasites, and white blood cells. Additionally, PI for platelet (PLT) components has the potential to extend PLT storage time from 5 to 7 days. A retrospective analysis was conducted into the percentage of outdated PLT components during the 3 years before and after the adoption of PLT PI technology in our institution. The PLT transfusion dose for both pre-PI and post-PI periods was similar. A retrospective analysis to study clinical safety and component utilization was also performed in the Balearic Islands University Hospital. As a result of PI implementation in our institution, the PLT production cost increased by 85.5%. However, due to the extension of PLT storage time, the percentage of outdated PLT units substantially decreased (-83.9%) and, consequently, the cost associated with outdated units (-69.8%). This decrease represented a 13.7% reduction of the initial cost increase which, together with the saving in blood transportation (0.1%), led to a saving of 13.8% over the initial cost. Therefore, the initial 85.5% increase in the cost of PLT production was markedly reduced to 71.7%. The mean number of PLT concentrates per patient was similar during both periods. The extension of PLT storage time can substantially contribute to reducing the financial impact of PI by decreasing the percentage of outdated PLTs while improving blood safety. Since the adoption of PI, there have been no documented cases of PLT transfusion-related sepsis in our region. © 2013 American Association of Blood Banks.
Elizabeth M. Powers; John D. Marshall; Jianwei Zhang; Liang Wei
2013-01-01
Forests mitigate climate change by sequestering CO2 from the atmosphere and accumulating it in biomass storage pools. However, in dry conifer forests, fire occasionally returns large quantities of CO2 to the atmosphere. Both the total amount of carbon stored and its susceptibility to loss may be altered by post-fire land...
2007-07-12
Nuclear Waste Storage Act of 2007. Requires commercial nuclear power plants to transfer spent fuel from pools to dry storage ...enrichment, spent fuel recycling (also called reprocessing), and other fuel cycle facilities that could be used to produce nuclear weapons materials...that had used the leased fuel , along with supplies of fresh nuclear fuel , according to the GNEP concept; see [http://www.gnep.energy.gov].
Meintker, Lisa; Haimerl, Maria; Ringwald, Jürgen; Krause, Stefan W
2013-11-01
Measurement of immature platelets was introduced into routine diagnostics by Sysmex as immature platelet fraction (IPF) some years ago and recently by Abbott as reticulated platelet fraction (rPT). Here, we compare both methods. We evaluated the precision and agreement of these parameters between Sysmex XE-5000 and Abbott CD-Sapphire in three distinct thrombocytopaenic cohorts: 30 patients with beginning thrombocytopaenia and 64 patients with recovering platelets (PLT) after chemotherapy, 16 patients with immune thrombocytopaenia (ITP) or heparin-induced thrombocytopaenia type 2 (HIT) and 110 additional normal controls. Furthermore, we analysed, how IPF/rPT differed between these thrombocytopaenic cohorts and controls. Both analysers demonstrated acceptable overall precision (repeatability) of IPF/rPT with lower precision at low PLT counts. IPF/rPT artificially increased during storage of blood samples overnight. Inter-instrument comparison showed a moderate correlation (Pearson r²=0.38) and a systematic bias of 1.04 towards higher IPF-values with the XE-5000. IPF/rPT was highest in recovering thrombopoesis after chemotherapy and moderately increased in ITP/HIT. The normal range deduced from control samples was much narrower with CD-Sapphire (1.0%-3.8%, established here for the first time) in comparison to XE-5000 (0.8%-7.9%) leading to a smaller overlap of samples with increased PLT turnover and normal controls. IPF and rPT both give useful information on PLT turnover, although the two analysers only show a moderate inter-instrument correlation and have different reference ranges. A better separation of patient groups with high PLT turnover like ITP/HIT from normal controls is obtained by CD-Sapphire.
Jurado, Marisa; Algora, Manuel; Garcia-Sanchez, Félix; Vico, Santiago; Rodriguez, Eva; Perez, Sonia; Barbolla, Luz
2012-01-01
Background The Community Transfusion Centre in Madrid currently processes whole blood using a conventional procedure (Compomat, Fresenius) followed by automated processing of buffy coats with the OrbiSac system (CaridianBCT). The Atreus 3C system (CaridianBCT) automates the production of red blood cells, plasma and an interim platelet unit from a whole blood unit. Interim platelet unit are pooled to produce a transfusable platelet unit. In this study the Atreus 3C system was evaluated and compared to the routine method with regards to product quality and operational value. Materials and methods Over a 5-week period 810 whole blood units were processed using the Atreus 3C system. The attributes of the automated process were compared to those of the routine method by assessing productivity, space, equipment and staffing requirements. The data obtained were evaluated in order to estimate the impact of implementing the Atreus 3C system in the routine setting of the blood centre. Yield and in vitro quality of the final blood components processed with the two systems were evaluated and compared. Results The Atreus 3C system enabled higher throughput while requiring less space and employee time by decreasing the amount of equipment and processing time per unit of whole blood processed. Whole blood units processed on the Atreus 3C system gave a higher platelet yield, a similar amount of red blood cells and a smaller volume of plasma. Discussion These results support the conclusion that the Atreus 3C system produces blood components meeting quality requirements while providing a high operational efficiency. Implementation of the Atreus 3C system could result in a large organisational improvement. PMID:22044958
Jurado, Marisa; Algora, Manuel; Garcia-Sanchez, Félix; Vico, Santiago; Rodriguez, Eva; Perez, Sonia; Barbolla, Luz
2012-01-01
The Community Transfusion Centre in Madrid currently processes whole blood using a conventional procedure (Compomat, Fresenius) followed by automated processing of buffy coats with the OrbiSac system (CaridianBCT). The Atreus 3C system (CaridianBCT) automates the production of red blood cells, plasma and an interim platelet unit from a whole blood unit. Interim platelet unit are pooled to produce a transfusable platelet unit. In this study the Atreus 3C system was evaluated and compared to the routine method with regards to product quality and operational value. Over a 5-week period 810 whole blood units were processed using the Atreus 3C system. The attributes of the automated process were compared to those of the routine method by assessing productivity, space, equipment and staffing requirements. The data obtained were evaluated in order to estimate the impact of implementing the Atreus 3C system in the routine setting of the blood centre. Yield and in vitro quality of the final blood components processed with the two systems were evaluated and compared. The Atreus 3C system enabled higher throughput while requiring less space and employee time by decreasing the amount of equipment and processing time per unit of whole blood processed. Whole blood units processed on the Atreus 3C system gave a higher platelet yield, a similar amount of red blood cells and a smaller volume of plasma. These results support the conclusion that the Atreus 3C system produces blood components meeting quality requirements while providing a high operational efficiency. Implementation of the Atreus 3C system could result in a large organisational improvement.
Culture of human cell lines by a pathogen-inactivated human platelet lysate.
Fazzina, R; Iudicone, P; Mariotti, A; Fioravanti, D; Procoli, A; Cicchetti, E; Scambia, G; Bonanno, G; Pierelli, L
2016-08-01
Alternatives to the use of fetal bovine serum (FBS) have been investigated to ensure xeno-free growth condition. In this study we evaluated the efficacy of human platelet lysate (PL) as a substitute of FBS for the in vitro culture of some human cell lines. PL was obtained by pools of pathogen inactivated human donor platelet (PLT) concentrates. Human leukemia cell lines (KG-1, K562, JURKAT, HL-60) and epithelial tumor cell lines (HeLa and MCF-7) were cultured with either FBS or PL. Changes in cell proliferation, viability, morphology, surface markers and cell cycle were evaluated for each cell line. Functional characteristics were analysed by drug sensitivity test and cytotoxicity assay. Our results demonstrated that PL can support growth and expansion of all cell lines, although the cells cultured in presence of PL experienced a less massive proliferation compared to those grown with FBS. We found a comparable percentage of viable specific marker-expressing cells in both conditions, confirming lineage fidelity in all cultures. Functionality assays showed that cells in both FBS- and PL-supported cultures maintained their normal responsiveness to adriamycin and NK cell-mediated lysis. Our findings indicate that PL is a feasible serum substitute for supporting growth and propagation of haematopoietic and epithelial cell lines with many advantages from a perspective of process standardization, ethicality and product safety.
Bordin, Antonella; Ponti, Donatella; Iudicone, Paola; Rendina, Erino Angelo; Calogero, Antonella; Pierelli, Luca; Ibrahim, Mohsen; De Falco, Elena
2015-01-01
Human adipose tissue-derived mesenchymal stem cells (ADMSCs) are considered eligible candidates for cardiovascular stem cell therapy applications due to their cardiac transdifferentiation potential and immunotolerance. Over the years, the in vitro culture of ADMSCs by platelet lysate (PL), a hemoderivate containing numerous growth factors and cytokines derived from platelet pools, has allowed achieving a safe and reproducible methodology to obtain high cell yield prior to clinical administration. Nevertheless, the biological properties of PL are still to be fully elucidated. In this brief report we show the potential ability of PL to induce a permissive state of cardiac-like transdifferentiation and to cause epigenetic modifications. RTPCR results indicate an upregulation of Cx43, SMA, c-kit, and Thy-1 confirmed by immunofluorescence staining, compared to standard cultures with foetal bovine serum. Moreover, PL-cultured ADMSCs exhibit a remarkable increase of both acetylated histones 3 and 4, with a patient-dependent time trend, and methylation at lysine 9 on histone 3 preceding the acetylation. Expression levels of p300 and SIRT-1, two major regulators of histone 3, are also upregulated after treatment with PL. In conclusion, PL could unravel novel biological properties beyond its routine employment in noncardiac applications, providing new insights into the plasticity of human ADMSCs. PMID:26495284
Siciliano, Camilla; Chimenti, Isotta; Bordin, Antonella; Ponti, Donatella; Iudicone, Paola; Peruzzi, Mariangela; Rendina, Erino Angelo; Calogero, Antonella; Pierelli, Luca; Ibrahim, Mohsen; De Falco, Elena
2015-01-01
Human adipose tissue-derived mesenchymal stem cells (ADMSCs) are considered eligible candidates for cardiovascular stem cell therapy applications due to their cardiac transdifferentiation potential and immunotolerance. Over the years, the in vitro culture of ADMSCs by platelet lysate (PL), a hemoderivate containing numerous growth factors and cytokines derived from platelet pools, has allowed achieving a safe and reproducible methodology to obtain high cell yield prior to clinical administration. Nevertheless, the biological properties of PL are still to be fully elucidated. In this brief report we show the potential ability of PL to induce a permissive state of cardiac-like transdifferentiation and to cause epigenetic modifications. RTPCR results indicate an upregulation of Cx43, SMA, c-kit, and Thy-1 confirmed by immunofluorescence staining, compared to standard cultures with foetal bovine serum. Moreover, PL-cultured ADMSCs exhibit a remarkable increase of both acetylated histones 3 and 4, with a patient-dependent time trend, and methylation at lysine 9 on histone 3 preceding the acetylation. Expression levels of p300 and SIRT-1, two major regulators of histone 3, are also upregulated after treatment with PL. In conclusion, PL could unravel novel biological properties beyond its routine employment in noncardiac applications, providing new insights into the plasticity of human ADMSCs.
Effect of Cold Storage on Shear-induced Platelet Aggregation and Clot Strength
2014-09-01
stored as 1-mL aliquots in 15-mL polypropylene centri- fuge tubes with gentle agitation either at RT (20Y24-C) or at 4-C (1Y6-C). Single AP units...vivo aging at 37-C. Brit J Haematol. 1995;91:212 218. 16. Babic AM, Josefsson EC, Bergmeier W, Wagner DD, Kaufman RM, Silberstein LE , et al. In vitro
A Civilian/Military Trauma Institute: National Trauma Coordinating Center
2016-12-01
temperature , higher inci- dence of both early (G6 hours) red blood cell transfusion and massive transfusion, higher mechanical ventilation require- ments, and... temperature on maintenance of platelet viabilityVdeleterious effect of refrigerated storage. N Engl J Med. 1969;280(20):1094Y1098. 9. Murphy DL, Colburn...Cohen, MD, University of California, San Francisco. b) Comparative Effectiveness of Clinical Care Processes in Resuscitation and Management of
Deng, Juhong; Zhang, Peng; Sun, Yue; Peng, Ping; Huang, Yu
2018-03-01
The prognostic and clinicopathological significance of the platelet to lymphocyte ratio (PLR) has been studied in various cancers. However, studies examining the role of PLR in esophageal cancer have not yielded consistent results. The purpose of this meta-analysis was to study the prognostic and clinicopathological significance of PLR in esophageal cancer patients. We performed a literature search in three major databases: PubMed, Web of Science and Embase (up until May 1, 2017). The clinicopathologic significance of PLR and its prognostic significance were analyzed. Our meta-analysis consisted of 13 studies with 4,621 patients. The pooled hazard ratios (HRs) showed that a high PLR was associated with poor survival of esophageal cancer [HR =1.283; 95% confidence interval (CI): 1.173-1.404; P<0.001]. Subgroup analysis revealed that elevated PLR was associated with poor survival in esophageal squamous cell carcinoma (HR =1.281; 95% CI: 1.098-1.493; P=0.002). The pooled odds ratio (OR) indicated that high PLR was also associated with the depth of tumor invasion (OR =1.543, 95% CI: 1.269-1.876, P<0.001), lymph node metastasis (OR =1.427, 95% CI: 1.195-1.705, P<0.001), tumor length (OR =1.81, 95% CI: 1.331-2.461, P<0.001), and Tumor stage (OR =1.459, 95% CI: 1.235-1.724, P<0.001). Our results demonstrate that elevated PLR was significantly associated with poor prognosis of esophageal cancer. Furthermore, the high PLR might predict worse clinicopathological features of esophageal cancer patients.
Gowland, Peter; Fontana, Stefano; Stolz, Martin; Andina, Nicola; Niederhauser, Christoph
2016-01-01
Summary Background Pathogen reduction methods for blood components are effective for a large number of viruses though less against small, non-enveloped viruses such as Parvovirus B19 (B19V). This article describes the passive transmission by transfusion of two B19V-contaminated pooled platelet concentrates (PCs) which were treated with the Intercept® blood pathogen reduction system. Case Reports Two transfusion cases of B19V-contaminated Intercept-treated pooled PCs were described. Due to the analysis delay, the PCs were already transfused. The viral content of each donation was 4.87 × 1010 IU/ml in case 1and 1.46 × 108 IU/ml in case 2. B19V (52 IU/ml) was detected in the recipient of the case 1 PC, whereas no virus could be detected in the case 2 PC recipient. A B19V IgM response and a transient boost of the underlying B19V IgG immune status and was observed in recipient 1. Recipient of the case 2 PC remained B19V IgG- and IgM-negative. B19V DNA sequence and phylogenetic analysis revealed a 100% homology between donor and recipient. Conclusion This report describes passive B19V transmission by a PC with very high B19 viral load which elicited a transient boost of the B19V immunity, but not by a PC with a lower B19V content, suggesting that there is a B19 viral load threshold value at which B19V inactivation is exceeded. PMID:27403092
Wei, Jing; Zhao, Jing-zhu; Deng, Hong-bing; Wu, Gang; Hao, Ying-jie; Shang, Wen-yan
2005-03-01
The nitrogen bio-cycle was discussed in the alpine tundra ecosystem of Changbai Mountain through compartment model. The alpine tundra of Changbai Mountain was compared with Arctic tundra by the common ratio of genus and species in this paper. It was found that the 89.3% of genus and 58.6% of species was the common between Changbai alpine tundra and Arctic tundra while 95.5% of lichen genus and 58.7% lichen species, 82.1% of moss genus and 76.3% of moss species, 93.1% of vascular bundle genus and 40.5% of vascular bundle species were the common, respectively, which made vegetation type or community to be similar between Changbai alpine tundra and Arctic tundra. The total storage of nitrogen was 65220.6 t in the vegetation-plant system of Changbai Mountain, of which soil pool amounted to 99.3%. The nitrogen storage of each compartment was as follows: the vegetation pool, litterfall pool and soil pool were 237.4 t, 145.3 t and 64837.9 t respectively. The transferable amounts of nitrogen were 131.7 t x a(-1), 58 t/a and 73.7 t x a(-1) in the aboveground plant, belowground root system and litterfall of alpine tundra ecosystem of Changbai Mountain.
van der Meer, Pieter F; Cancelas, Jose A; Cardigan, Rebecca; Devine, Dana V; Gulliksson, Hans; Sparrow, Rosemary L; Vassallo, Ralph R; de Wildt-Eggen, Janny; Baumann-Baretti, Bärbel; Hess, John R
2011-01-01
Whole blood (WB) can be held at room temperature (18-25°C) up to 8 hours after collection; thereafter the unit must be refrigerated, rendering it unsuitable for platelet (PLT) production. Overnight hold at room temperature before processing has logistic advantages, and we evaluated this process in an international multicenter study for both buffy coat (BC)- and PLT-rich plasma (PRP)-based blood components and compared three red blood cell (RBC) additive solutions (ASs) for their ability to offset effects of overnight hold. Nine centers participated; seven used the BC method, and two used the PRP method. Four WB units were pooled and split; 1 unit was processed less than 8 hours from collection (Group A), and the other three (Groups B, C, and D) were held at room temperature and processed after 24 to 26 hours. RBCs in Groups A and B were resuspended in saline-adenine-glucose-mannitol, Group C in phosphate-adenine-guanosine-glucose-saline-mannitol, and Group D in ErythroSol-4 RBCs were stored at 2 to 6°C for 49 days. RBCs from overnight-held WB had lower 2,3-diphosphoglycerate (2,3-DPG) and higher adenosine triphosphate (ATP). At the end of storage there were no differences between groups, apart from a slightly higher hemolysis in Group B. ErythroSol-4 showed a slightly higher initial ATP and 2,3-DPG content, but at the end of storage no differences were found. Overnight hold of WB before processing has no lasting deleterious effects on in vitro quality of subsequently prepared components. The use of different RBC ASs did not appear to offer significant advantages in terms of RBC quality at the end, regardless of the processing method. © 2010 American Association of Blood Banks.
Sediment transport through self-adjusting, bedrock-walled waterfall plunge pools
NASA Astrophysics Data System (ADS)
Scheingross, Joel S.; Lamb, Michael P.
2016-05-01
Many waterfalls have deep plunge pools that are often partially or fully filled with sediment. Sediment fill may control plunge-pool bedrock erosion rates, partially determine habitat availability for aquatic organisms, and affect sediment routing and debris flow initiation. Currently, there exists no mechanistic model to describe sediment transport through waterfall plunge pools. Here we develop an analytical model to predict steady-state plunge-pool depth and sediment-transport capacity by combining existing jet theory with sediment transport mechanics. Our model predicts plunge-pool sediment-transport capacity increases with increasing river discharge, flow velocity, and waterfall drop height and decreases with increasing plunge-pool depth, radius, and grain size. We tested the model using flume experiments under varying waterfall and plunge-pool geometries, flow hydraulics, and sediment size. The model and experiments show that through morphodynamic feedbacks, plunge pools aggrade to reach shallower equilibrium pool depths in response to increases in imposed sediment supply. Our theory for steady-state pool depth matches the experiments with an R2 value of 0.8, with discrepancies likely due to model simplifications of the hydraulics and sediment transport. Analysis of 75 waterfalls suggests that the water depths in natural plunge pools are strongly influenced by upstream sediment supply, and our model provides a mass-conserving framework to predict sediment and water storage in waterfall plunge pools for sediment routing, habitat assessment, and bedrock erosion modeling.
Military walking blood bank and the civilian blood service.
Berséus, Olle; Hervig, Tor; Seghatchian, Jerard
2012-06-01
In most countries whole blood transfusions have been replaced by component therapy. This has allowed for both better usage of the blood donations and better quality during storage. While this strategy was initially motivated by the commercial need for plasma the plasma reduction also reduced the levels of low grade proteases and sialidase, hence minimizing the cellular storage lesion/microvesiculation during prolonged storage. Plasma reduction also reduces transfusion reactions associated with plasma. During special military conditions, however, blood transfusion is urgently needed without corresponding access to blood components, in particular platelets. Accordingly, new focus on whole blood has aroused and added a new challenge to the blood transfusion services. This special issue of "what is happening" highlights the planed efforts by Swedish and Norwegian groups in the developments of military walking blood bank, which is applicable to civil blood services. Copyright © 2012 Elsevier Ltd. All rights reserved.
Osmotically inactive sodium and potassium storage: lessons learned from the Edelman and Boling data.
Nguyen, Minhtri K; Nguyen, Dai-Scott; Nguyen, Minh-Kevin
2016-09-01
Because changes in the plasma water sodium concentration ([Na(+)]pw) are clinically due to changes in the mass balance of Na(+), K(+), and H2O, the analysis and treatment of the dysnatremias are dependent on the validity of the Edelman equation in defining the quantitative interrelationship between the [Na(+)]pw and the total exchangeable sodium (Nae), total exchangeable potassium (Ke), and total body water (TBW) (Edelman IS, Leibman J, O'Meara MP, Birkenfeld LW. J Clin Invest 37: 1236-1256, 1958): [Na(+)]pw = 1.11(Nae + Ke)/TBW - 25.6. The interrelationship between [Na(+)]pw and Nae, Ke, and TBW in the Edelman equation is empirically determined by accounting for measurement errors in all of these variables. In contrast, linear regression analysis of the same data set using [Na(+)]pw as the dependent variable yields the following equation: [Na(+)]pw = 0.93(Nae + Ke)/TBW + 1.37. Moreover, based on the study by Boling et al. (Boling EA, Lipkind JB. 18: 943-949, 1963), the [Na(+)]pw is related to the Nae, Ke, and TBW by the following linear regression equation: [Na(+)]pw = 0.487(Nae + Ke)/TBW + 71.54. The disparities between the slope and y-intercept of these three equations are unknown. In this mathematical analysis, we demonstrate that the disparities between the slope and y-intercept in these three equations can be explained by how the osmotically inactive Na(+) and K(+) storage pool is quantitatively accounted for. Our analysis also indicates that the osmotically inactive Na(+) and K(+) storage pool is dynamically regulated and that changes in the [Na(+)]pw can be predicted based on changes in the Nae, Ke, and TBW despite dynamic changes in the osmotically inactive Na(+) and K(+) storage pool. Copyright © 2016 the American Physiological Society.
Federal Register 2010, 2011, 2012, 2013, 2014
2012-03-30
... consist of the following: (1) An upper reservoir formed by a 160-foot-high by 6,750-foot-long, roller- compacted concrete (RCC) dam (an open ``U''--shaped structure varying from grade to roughly 160-foot-high) having a total storage capacity of 5,530 acre-feet and a water surface area of 90 acres at full pool...
Influence of prescribed fire on ecosystem biomass, carbon, and nitrogen in a pinyon juniper woodland
Benjamin M. Rau; Robin Tausch; Alicia Reiner; Dale W. Johnson; Jeanne C. Chambers; Robert R. Blank; Annmarrie Lucchesi
2010-01-01
Increases in pinyon and juniper woodland cover associated with land-use history are suggested to provide offsets for carbon emissions in arid regions. However, the largest pools of carbon in arid landscapes are typically found in soils, and aboveground biomass cannot be considered long-term storage in fire-prone ecosystems. Also, the objectives of carbon storage may...
Federal Register 2010, 2011, 2012, 2013, 2014
2011-04-28
... seasonal flash boards; (2) a 168-acre reservoir with a gross storage capacity of 3,234 acre-feet and a...; (2) a 159-acre reservoir with a gross storage capacity of 2,646 acre-feet and a normal maximum pool... reservoir, with concrete core walls and partially equipped with 10-inch-high flashboards; (3) a 79.2- acre...
... and Women with Hemophilia Inheritance of Hemophilia Definitions & Terminology Bleeding Symptoms Carrier Diagnosis When to Test for ... and Women with Hemophilia Inheritance of Hemophilia Definitions & Terminology Bleeding Symptoms Carrier Diagnosis When to Test for ...
E.S. Kane; J.G. Vogel
2009-01-01
To understand how carbon (C) pools in boreal ecosystems may change with warming, we measured above- and belowground C pools and C increment along a soil temperature gradient across 16 mature upland black spruce (Picea mariana Mill. [Bâ¢S.P]) forests in interior Alaska. Total spruce C stocks (stand and root C) increased from 1.3 to 8.5 kg C m
NASA Technical Reports Server (NTRS)
Soltis, Steven R.; Ruwart, Thomas M.; OKeefe, Matthew T.
1996-01-01
The global file system (GFS) is a prototype design for a distributed file system in which cluster nodes physically share storage devices connected via a network-like fiber channel. Networks and network-attached storage devices have advanced to a level of performance and extensibility so that the previous disadvantages of shared disk architectures are no longer valid. This shared storage architecture attempts to exploit the sophistication of storage device technologies whereas a server architecture diminishes a device's role to that of a simple component. GFS distributes the file system responsibilities across processing nodes, storage across the devices, and file system resources across the entire storage pool. GFS caches data on the storage devices instead of the main memories of the machines. Consistency is established by using a locking mechanism maintained by the storage devices to facilitate atomic read-modify-write operations. The locking mechanism is being prototyped in the Silicon Graphics IRIX operating system and is accessed using standard Unix commands and modules.
Estrogen Receptor Mutants/Variants in Human Breast Cancer.
1995-12-01
Effect of dystrophin gene deletions on mRNA levels and processing in Duchenne and Becker dystrophies . Cell 1990, 63:1239-1248 4 Patriotis C, Makris A...the most common inherited disorder of platelets, Aspartylblucosaminuria, 2 an inherited lysosomal storage disorder, Duchenne and Becker muscular... dystrophies 3 or cancer progression.4m 5 Several estrogen receptor (ER) variant mRNAs have also been identified in human breast cancer biopsies.6,7,8, 9
Effects of Lower Drying-Storage Temperature on the Ductility of High-Burnup PWR Cladding
DOE Office of Scientific and Technical Information (OSTI.GOV)
Billone, M. C.; Burtseva, T. A.
2016-08-30
The purpose of this research effort is to determine the effects of canister and/or cask drying and storage on radial hydride precipitation in, and potential embrittlement of, high-burnup (HBU) pressurized water reactor (PWR) cladding alloys during cooling for a range of peak drying-storage temperatures (PCT) and hoop stresses. Extensive precipitation of radial hydrides could lower the failure hoop stresses and strains, relative to limits established for as-irradiated cladding from discharged fuel rods stored in pools, at temperatures below the ductile-to-brittle transition temperature (DBTT).
Effects of Management on Soil Carbon Pools in California Rangeland Ecosystems
NASA Astrophysics Data System (ADS)
Silver, W. L.; Ryals, R.; Lewis, D. J.; Creque, J.; Wacker, M.; Larson, S.
2008-12-01
Rangeland ecosystems managed for livestock production represent the largest land-use footprint globally, covering more than one-quarter of the world's land surface (Asner et al. 2004). In California, rangelands cover an estimated 17 million hectares or approximately 40% of the land area (FRAP 2003). These ecosystems have considerable potential to sequester carbon (C) in soil and offset greenhouse gas emissions through changes in land management practices. Climate policies and C markets may provide incentives for rangeland managers to pursue strategies that optimize soil C storage, yet we lack a thorough understanding of the effects of management on soil C pools in rangelands over time and space. We sampled soil C pools on rangelands in a 260 km2 region of Marin and Sonoma counties to determine if patterns in soil C storage exist with management. Replicate soil samples were collected from 35 fields that spanned the dominant soil orders, plant communities, and management practices in the region while controlling for slope and bioclimatic zone (n = 1050). Management practices included organic amendments, intensive (dairy) and extensive (other) grazing practices, and subsoiling. Soil C pools ranged from approximately 50 to 140 Mg C ha-1 to 1 m depth, with a mean of 99 ± 22 (sd) Mg C ha-1. Differences among sites were due primarily to C concentrations, which exhibited a much larger coefficient of variation than bulk density at all depths. There were no statistically significant differences among the dominant soil orders. Subsoiling appeared to significantly increase soil C content in the top 50 cm, even though subsoiling had only occurred for the first time the previous Nov. Organic amendments also appeared to greatly increase soil C pools, and was the dominant factor that distinguished soil C pools in intensive and extensive land uses. Our results indicate that management has the potential to significantly increase soil C pools. Future research will determine the location of sequestered C within the soil matrix and its turnover time.
Wei, Yawei; Li, Maihe; Chen, Hua; Lewis, Bernard J; Yu, Dapao; Zhou, Li; Zhou, Wangming; Fang, Xiangmin; Zhao, Wei; Dai, Limin
2013-01-01
The northeastern forest region of China is an important component of total temperate and boreal forests in the northern hemisphere. But how carbon (C) pool size and distribution varies among tree, understory, forest floor and soil components, and across stand ages remains unclear. To address this knowledge gap, we selected three major temperate and two major boreal forest types in northeastern (NE) China. Within both forest zones, we focused on four stand age classes (young, mid-aged, mature and over-mature). Results showed that total C storage was greater in temperate than in boreal forests, and greater in older than in younger stands. Tree biomass C was the main C component, and its contribution to the total forest C storage increased with increasing stand age. It ranged from 27.7% in young to 62.8% in over-mature stands in boreal forests and from 26.5% in young to 72.8% in over-mature stands in temperate forests. Results from both forest zones thus confirm the large biomass C storage capacity of old-growth forests. Tree biomass C was influenced by forest zone, stand age, and forest type. Soil C contribution to total forest C storage ranged from 62.5% in young to 30.1% in over-mature stands in boreal and from 70.1% in young to 26.0% in over-mature in temperate forests. Thus soil C storage is a major C pool in forests of NE China. On the other hand, understory and forest floor C jointly contained less than 13% and <5%, in boreal and temperate forests respectively, and thus play a minor role in total forest C storage in NE China.
Wei, Yawei; Li, Maihe; Chen, Hua; Lewis, Bernard J.; Yu, Dapao; Zhou, Li; Zhou, Wangming; Fang, Xiangmin; Zhao, Wei; Dai, Limin
2013-01-01
The northeastern forest region of China is an important component of total temperate and boreal forests in the northern hemisphere. But how carbon (C) pool size and distribution varies among tree, understory, forest floor and soil components, and across stand ages remains unclear. To address this knowledge gap, we selected three major temperate and two major boreal forest types in northeastern (NE) China. Within both forest zones, we focused on four stand age classes (young, mid-aged, mature and over-mature). Results showed that total C storage was greater in temperate than in boreal forests, and greater in older than in younger stands. Tree biomass C was the main C component, and its contribution to the total forest C storage increased with increasing stand age. It ranged from 27.7% in young to 62.8% in over-mature stands in boreal forests and from 26.5% in young to 72.8% in over-mature stands in temperate forests. Results from both forest zones thus confirm the large biomass C storage capacity of old-growth forests. Tree biomass C was influenced by forest zone, stand age, and forest type. Soil C contribution to total forest C storage ranged from 62.5% in young to 30.1% in over-mature stands in boreal and from 70.1% in young to 26.0% in over-mature in temperate forests. Thus soil C storage is a major C pool in forests of NE China. On the other hand, understory and forest floor C jointly contained less than 13% and <5%, in boreal and temperate forests respectively, and thus play a minor role in total forest C storage in NE China. PMID:23977252
Cost implications of implementation of pathogen-inactivated platelets
McCullough, Jeffrey; Goldfinger, Dennis; Gorlin, Jed; Riley, William J; Sandhu, Harpreet; Stowell, Christopher; Ward, Dawn; Clay, Mary; Pulkrabek, Shelley; Chrebtow, Vera; Stassinopoulos, Adonis
2015-01-01
BACKGROUND Pathogen inactivation (PI) is a new approach to blood safety that may introduce additional costs. This study identifies costs that could be eliminated, thereby mitigating the financial impact. STUDY DESIGN AND METHODS Cost information was obtained from five institutions on tests and procedures (e.g., irradiation) currently performed, that could be eliminated. The impact of increased platelet (PLT) availability due to fewer testing losses, earlier entry into inventory, and fewer outdates with a 7-day shelf life were also estimated. Additional estimates include costs associated with managing 1) special requests and 2) test results, 3) quality control and proficiency testing, 4) equipment acquisition and maintenance, 5) replacement of units lost to positive tests, 6) seasonal or geographic testing, and 7) health department interactions. RESULTS All costs are mean values per apheresis PLT unit in USD ($/unit). The estimated test costs that could be eliminated are $71.76/unit and a decrease in transfusion reactions corresponds to $2.70/unit. Avoiding new tests (e.g., Babesia and dengue) amounts to $41.80/unit. Elimination of irradiation saves $8.50/unit, while decreased outdating with 7-day storage can be amortized to $16.89/unit. Total potential costs saved with PI is $141.65/unit. Costs are influenced by a variety of factors specific to institutions such as testing practices and the location in which such costs are incurred and careful analysis should be performed. Additional benefits, not quantified, include retention of some currently deferred donors and scheduling flexibility due to 7-day storage. CONCLUSIONS While PI implementation will result in additional costs, there are also potential offsetting cost reductions, especially after 7-day storage licensing. PMID:25989465
Age of platelet concentrates and time to the next transfusion.
Caram-Deelder, Camila; van der Bom, Johanna G; Putter, Hein; Leyte, Anja; Kerkhof, Daan van de; Evers, Dorothea; Beckers, Erik A; Weerkamp, Floor; Hudig, Francisca; Zwaginga, Jaap Jan; Rondeel, Jan M M; de Vooght, Karen M K; Péquériaux, Nathalie C V; Visser, Otto; Wallis, Jonathan P; Middelburg, Rutger A
2018-01-01
Storage time of platelet (PLT) concentrates has been negatively associated with clinical efficacy outcomes. The aim of this study was to quantify the association between storage time of PLT concentrates and interval to the next PLT transfusion for different types of PLT components, stored for up to 7 days and transfused to transfusion-dependent hematooncology patients with thrombocytopenia. From a cohort of patients from 10 major Dutch hospitals, patients were selected whose transfusion patterns were compatible with PLT transfusion dependency due to hematooncologic disease. Mean time to the next transfusion and mean differences in time to the next transfusion for different storage time categories (i.e., fresh, <4 days; intermediate, 4-5 days; and old, >5 days) were estimated, per component type, using multilevel mixed-effects linear models. Among a cohort of 29,761 patients who received 140,896 PLT transfusions we selected 4441 hematooncology patients who had received 12,724 PLT transfusions during periods of PLT transfusion dependency. Transfusion of fresh, compared to old, buffy coat-derived PLTs in plasma was associated with a delay to the next transfusion of 6.2 hours (95% confidence interval [CI], 4.5-8.0 hr). For buffy coat-derived PLTs in PAS-B and -C this difference was 7.7 hours (95% CI, 2.2-13.3 hr) and 3.9 hours (95% CI, -2.1 to 9.9 hr) while for apheresis PLTs in plasma it was only 1.8 hours (95% CI, -3.5 to 7.1 hr). Our results indicate that the time to the next transfusion shortens with increasing age of transfused buffy coat-derived PLT concentrates. This association was not observed for apheresis PLTs. © 2017 AABB.
Cost implications of implementation of pathogen-inactivated platelets.
McCullough, Jeffrey; Goldfinger, Dennis; Gorlin, Jed; Riley, William J; Sandhu, Harpreet; Stowell, Christopher; Ward, Dawn; Clay, Mary; Pulkrabek, Shelley; Chrebtow, Vera; Stassinopoulos, Adonis
2015-10-01
Pathogen inactivation (PI) is a new approach to blood safety that may introduce additional costs. This study identifies costs that could be eliminated, thereby mitigating the financial impact. Cost information was obtained from five institutions on tests and procedures (e.g., irradiation) currently performed, that could be eliminated. The impact of increased platelet (PLT) availability due to fewer testing losses, earlier entry into inventory, and fewer outdates with a 7-day shelf life were also estimated. Additional estimates include costs associated with managing (1) special requests and (2) test results, (3) quality control and proficiency testing, (4) equipment acquisition and maintenance, (5) replacement of units lost to positive tests, (6) seasonal or geographic testing, and (7) health department interactions. All costs are mean values per apheresis PLT unit in USD ($/unit). The estimated test costs that could be eliminated are $71.76/unit and a decrease in transfusion reactions corresponds to $2.70/unit. Avoiding new tests (e.g., Babesia and dengue) amounts to $41.80/unit. Elimination of irradiation saves $8.50/unit, while decreased outdating with 7-day storage can be amortized to $16.89/unit. Total potential costs saved with PI is $141.65/unit. Costs are influenced by a variety of factors specific to institutions such as testing practices and the location in which such costs are incurred and careful analysis should be performed. Additional benefits, not quantified, include retention of some currently deferred donors and scheduling flexibility due to 7-day storage. While PI implementation will result in additional costs, there are also potential offsetting cost reductions, especially after 7-day storage licensing. © 2015 The Authors Transfusion published by Wiley Periodicals, Inc. on behalf of AABB.
Soil organic carbon and total nitrogen pools in permafrost zones of the Qinghai-Tibetan Plateau.
Zhao, Lin; Wu, Xiaodong; Wang, Zhiwei; Sheng, Yu; Fang, Hongbing; Zhao, Yonghua; Hu, Guojie; Li, Wangping; Pang, Qiangqiang; Shi, Jianzong; Mo, Bentian; Wang, Qian; Ruan, Xirui; Li, Xiaodong; Ding, Yongjian
2018-02-26
There are several publications related to the soil organic carbon (SOC) on the Qinghai-Tibetan Plateau (QTP). However, most of these reports were from different parts of the plateau with various sampling depth. Here, we present the results from a systematic sampling and analysis of 200 soil pits. Most of the pits were deeper than 2 m from an east-west transect across the plateau. The SOC and total nitrogen (TN) pools of the 148 × 10 4 km 2 , the area of the permafrost zone, for the upper 2 m soils calculated from the vegetation map were estimated to be 17.07 Pg (interquartile range: 11.34-25.33 Pg) and 1.72 Pg (interquartile range: 1.08-2.06 Pg), respectively. We also predicted the distribution of land cover types in 2050 and 2070 using decision tree rules and climate scenarios, and then predicted SOC and TN pools of this region. The results suggested that the SOC and TN pools will decrease in the future. The results not only contribute to the carbon and nitrogen storage and stocks in the permafrost regions as a whole but most importantly, to our knowledge of the possible changes of C and N storage on the QTP in the future.
Fine-grained sediment storage conditioned by Large Woody Debris in a gravel-bed river
NASA Astrophysics Data System (ADS)
Skalak, K. J.; Narinesingh, P.; Pizzuto, J. E.
2006-05-01
The purposes of this study are 1) to determine the quantity of mud and sand stored in the channel margins and near-bank regions of South River, a steep gravel-bedded stream in western Virginia, and 2) to understand the geomorphic and hydrologic processes that control the erosion and deposition of these fine-grained deposits. The volume of storage in these deposits is equivalent to about 5-10 percent of the river's annual suspended sediment load. Sediment storage in the near-bank regions is a result of reduced velocity caused by the bank obstructions. Storage occurs in four different geomorphic settings: 1) long pooled sections caused by bedrock or old mill dams, 2) regions downstream of riffles in channel margins with LWD accumulations, 3) bank obstructions usually caused by trees, 4) side channel backwaters where flow separates around islands. Most storage occurs in regions downstream of riffles (approximately 44 percent of the total). Long pooled sections account for roughly 37 percent of the total storage, bank obstructions account for 13 percent, and backwaters account for roughly 6 percent. In approximately 17 km of river, there are 38 separate fine-grained deposits (total volume more than 1600 m3). On average, these deposits are about 35 cm deep, 20 m long, and 4 m wide. They average 30 percent mud, 68 percent sand, and 2 percent gravel. These deposits have been cored and analyzed for Hg, grain size, loss-on-ignition, and bomb radiocarbon. High Hg concentrations in fish tissue are an ongoing problem along South River, further motivating detailed study of these deposits.
NASA Astrophysics Data System (ADS)
Telles, Everaldo De Carvalho ConceiçÃ.£O.; de Camargo, PlíNio Barbosa; Martinelli, Luiz A.; Trumbore, Susan E.; da Costa, Enir Salazar; Santos, Joaquim; Higuchi, Niro; Oliveira, Raimundo Cosme
2003-06-01
Stable and radiocarbon isotopes were used to investigate the role of soil clay content in the storage and dynamics of soil carbon in tropical forest soils. Organic matter in clay-rich Oxisols and Ultisols contains at least two distinct components: (1) material with light δ13C signatures and turnover times of decades or less; and (2) clay-associated, 13C-enriched, carbon with turnover times of decades at the surface to millennia at depths >20 cm. Soil texture, in this case clay content, exerts a major control on the amount of slowly cycling carbon and therefore influences the storage and dynamics of carbon in tropical forest soils. Soils in primary tropical forest have been proposed as a potentially large sink for anthropogenic carbon. Comparison of carbon stocks in Oxisols sampled near Manaus, Brazil, shows no measurable change in organic carbon stocks over the past 20 years. Simple models estimating the response of soil carbon pools to a sustained 0.5% yr-1 increase in productivity result in C storage rates of 0.09 to 0.13 MgC ha-1 yr-1 in soil organic matter, with additional potential storage of 0.18 to 0.27 MgC ha-1 yr-1 in surface litter and roots. Most storage occurs in organic matter pools with turnover times less than a decade. Export of carbon in dissolved form from upland terra firme Oxisols likely accounts for <0.2 MgC ha-1 yr-1, but more work is required to assess the export potential for periodically inundated Spodosols.
de Korte, Dirk; Thibault, Louis; Handke, Wiebke; Harm, Sarah K; Morrison, Alex; Fitzpatrick, Aine; Marks, Denese C; Yi, Qi-Long; Acker, Jason P
2018-04-01
There are few studies investigating the effect of irradiation on red blood cells (RBCs) during storage. This study analyzed changes in in vitro quality of RBCs irradiated at several points during storage with the aim of providing evidence to support current maximum pre- and postirradiation storage limits. Each of seven participating centers produced four pools of 7 standard RBC units (SAGM, AS-3, or PAGGSM), which were then split back into 7 units. All units in a pool were from sex-matched blood donors. Every week during 6 weeks of refrigerated storage, 1 unit was irradiated, while 1 unit was not irradiated (control). Units were tested weekly for biochemical variables, morphology, and mechanical fragility. The earlier during storage that units were irradiated, the higher the hemolysis and K + at end of storage. Irrespective of the timing of irradiation, there was a rapid increase in extracellular K + , followed by a more gradual increase in hemolysis. ATP levels decreased faster in irradiated units and were reduced below accepted values if irradiated early. Irradiated female RBCs had an absolute lower hemolysis and K + level compared to male RBCs at all time points. The method of blood component manufacturing determined the absolute levels of hemolysis and potassium in irradiated and nonirradiated units, but did not influence the effect that timing of irradiation had on the in vitro quality characteristics. This study provides support for the current Council of Europe guidelines on the time limitations for the irradiation of RBCs. © 2017 AABB.
NASA Astrophysics Data System (ADS)
Vogel, J. G.; Bacon, A. R.; Bracho, R. G.; Gonzalez-Benecke, C. A.; Fox, T. D.; Laviner, M. A.; Kane, M.; Burkhart, H.; Martin, T.; Will, R.; Ross, C. W.; Grunwald, S.; Jokela, E. J.; Meek, C.
2016-12-01
Extending from Virginia to east Texas in the southeastern United States, managed pine plantations are an important component of the region's carbon cycle. An objective of the Pine Integrated Network: Education, Mitigation, and Adaptation project (PINEMAP) is to improve estimates of how ecosystem carbon pools respond to the management strategies used to increase the growth of loblolly pine plantations. Experimental studies (108 total) that have been used to understand plantation productivity and stand dynamics by university-forest industry cooperatives were measured for the carbon stored in the trees, roots, coarse-wood, detritus in soil, forest floor, understory and soils to 1-meter. The age of the studied plantations ranged from 4-26 years at the time of sampling, with 26 years very near the period when these plantations are commonly harvested. Across all study sites, 455 experimental plots were measured. The average C storage across all pools, sites, and treatments was 192 Mg C ha-1, with the average percentage of the total coming from soil (44%), tree biomass (40%), forest floor (8%), root (5%), soil detritus (2%), understory biomass (1%), and coarse-wood (<1%) pools. Plots had as a treatment either fertilization, competition control, and stand density control (thinning), and every possible combination of treatments including `no treatment'. A paired plot analysis was used where two plots at a site were examined for relative differences caused by a single treatment and these differences averaged across the region. Thinning as a stand-alone treatment significantly reduced forest floor mass by 60%, and the forest floor in the thinned plus either competition control or fertilization was 18.9% and 19.2% less, respectively, than unthinned stands combined with the same treatments. Competition control increased C storage in tree biomass by 12% and thinning decreased tree biomass by 32%. Thinning combined with fertilization had lower soil carbon (0-1 m) than unthinned-fertilized plots (22%), although the replication for this combination was relatively low (n=6). Overall these results suggest that maintaining higher tree densities increases ecosystem carbon storage across multiple pools of C in loblolly pine plantations.
Canine and feline blood transfusions: controversies and recent advances in administration practices.
Kisielewicz, Caroline; Self, Ian A
2014-05-01
To discuss and review blood transfusion practices in dogs and cats including collection and storage of blood and administration of products. To report new developments, controversial practices, less conventional blood product administration techniques and where applicable, describe the relevance to anaesthetists and anaesthesia. PubMed and Google Scholar using dog, cat, blood transfusion, packed red blood cells and whole blood as keywords. Blood transfusions improve oxygen carrying capacity and the clinical signs of anaemia. However there are numerous potential risks and complications possible with transfusions, which may outweigh their benefits. Storage of blood products has improved considerably over time but whilst extended storage times may improve their availability, a phenomenon known as the storage lesion has been identified which affects erythrocyte viability and survival. Leukoreduction involves removing leukocytes and platelets thereby preventing their release of cytokines and bioactive compounds which also contribute to storage lesions and certain transfusion reactions. Newer transfusion techniques are being explored such as cell salvage in surgical patients and subsequent autologous transfusion. Xenotransfusions, using blood and blood products between different species, provide an alternative to conventional blood products. © 2014 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia.
Chahal, Jaskarndip; Van Thiel, Geoffrey S; Mall, Nathan; Heard, Wendell; Bach, Bernard R; Cole, Brian J; Nicholson, Gregory P; Verma, Nikhil N; Whelan, Daniel B; Romeo, Anthony A
2012-11-01
Despite the theoretic basis and interest in using platelet-rich plasma (PRP) to improve the potential for rotator cuff healing, there remains ongoing controversy regarding its clinical efficacy. The objective of this systematic review was to identify and summarize the available evidence to compare the efficacy of arthroscopic rotator cuff repair in patients with full-thickness rotator cuff tears who were concomitantly treated with PRP. We searched the Cochrane Central Register of Controlled Trials, Medline, Embase, and PubMed for eligible studies. Two reviewers selected studies for inclusion, assessed methodologic quality, and extracted data. Pooled analyses were performed using a random effects model to arrive at summary estimates of treatment effect with associated 95% confidence intervals. Five studies (2 randomized and 3 nonrandomized with comparative control groups) met the inclusion criteria, with a total of 261 patients. Methodologic quality was uniformly sound as assessed by the Detsky scale and Newcastle-Ottawa Scale. Quantitative synthesis of all 5 studies showed that there was no statistically significant difference in the overall rate of rotator cuff retear between patients treated with PRP and those treated without PRP (risk ratio, 0.77; 95% confidence interval, 0.48 to 1.23). There were also no differences in the pooled Constant score; Simple Shoulder Test score; American Shoulder and Elbow Surgeons score; University of California, Los Angeles shoulder score; or Single Assessment Numeric Evaluation score. PRP does not have an effect on overall retear rates or shoulder-specific outcomes after arthroscopic rotator cuff repair. Additional well-designed randomized trials are needed to corroborate these findings. Level III, systematic review of Level I, II, and III studies. Copyright © 2012 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.
Tchang, Laurent A; Pippenger, Benjamin E; Todorov, Atanas; Wolf, Francine; Burger, Maximilian G; Jaquiery, Claude; Bieback, Karen; Martin, Ivan; Schaefer, Dirk J; Scherberich, Arnaud
2017-05-01
The use of fetal bovine serum (FBS) as a culture medium supplement in cell therapy and clinical tissue engineering is challenged by immunological concerns and the risk of disease transmission. Here we tested whether human, thrombin-activated, pooled, platelet-rich plasma (tPRP) can be substituted for FBS in the engineering of osteogenic and vasculogenic grafts, using cells from the stromal vascular fraction (SVF) of human adipose tissue. SVF cells were cultured under perfusion flow into porous hydroxyapatite scaffolds for 5 days, with the medium supplemented with either 10% tPRP or 10% FBS and implanted in an ectopic mouse model. Following in vitro culture, as compared to FBS, the use of tPRP did not modify the fraction of clonogenic cells or the different cell phenotypes, but increased by 1.9-fold the total number of cells. After 8 weeks in vivo, bone tissue was formed more reproducibly and in higher amounts (3.7-fold increase) in constructs cultured with tPRP. Staining for human-specific ALU sequences and for the human isoforms of CD31/CD34 revealed the human origin of the bone, the formation of blood vessels by human vascular progenitors and a higher density of human cells in implants cultured with tPRP. In summary, tPRP supports higher efficiency of bone formation by SVF cells than FBS, likely by enhancing cell expansion in vitro while maintaining vasculogenic properties. The use of tPRP may facilitate the clinical translation of osteogenic grafts with intrinsic capacity for vascularization, based on the use of adipose-derived cells. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.
Horizontal modular dry irradiated fuel storage system
Fischer, Larry E.; McInnes, Ian D.; Massey, John V.
1988-01-01
A horizontal, modular, dry, irradiated fuel storage system (10) includes a thin-walled canister (12) for containing irradiated fuel assemblies (20), which canister (12) can be positioned in a transfer cask (14) and transported in a horizontal manner from a fuel storage pool (18), to an intermediate-term storage facility. The storage system (10) includes a plurality of dry storage modules (26) which accept the canister (12) from the transfer cask (14) and provide for appropriate shielding about the canister (12). Each module (26) also provides for air cooling of the canister (12) to remove the decay heat of the irradiated fuel assemblies (20). The modules (26) can be interlocked so that each module (26) gains additional shielding from the next adjacent module (26). Hydraulic rams (30) are provided for inserting and removing the canisters (12) from the modules (26).
Fresh Whole Blood Use for Hemorrhagic Shock: Preserving Benefit While Avoiding Complications
2012-10-01
coagulation factor deficiency), and improved blood bank economics.1–3 However, component usage in austere settings (developing world and combat) is often...limited by storage requirements for blood products (refrig- eration for red blood cells [RBCs], room temperature with agitation for platelets [PLTs...red blood cell (RBC) or plasma product inven- tory” (Ref. 1 in Table 1). Most civilian institutions define it as fresh if it is stored for 48 hours
Noninvasive pH monitoring of platelet concentrates: a large field test.
Gkoumassi, Effimia; Klein-Bosgoed, Christa; Dijkstra-Tiekstra, Margriet J; de Korte, Dirk; de Wildt-Eggen, Janny
2013-10-01
Developing new quality control methods for platelet concentrates (PCs) can contribute to increasing transfusion safety and efficiency. The aim of this study was to investigate in a large field test the quality of expired PCs and whether 100% noninvasive pH monitoring can be used to predict PC quality. The pH of 13,693 PCs produced for transfusion was monitored daily using Blood Storage, Inc.'s pH sterile, automated fluoroscopic evaluation technology. Upon indication of compromised quality or expiration, PCs were returned and in vitro tests were performed. A total of 998 PCs were returned, of which 962 outdated, 26 had a positive BacT/ALERT reaction, seven had aggregates, one was without swirl, one had low pH, and one had high pH. BacT/ALERT was faster in identifying bacterial contamination than pH measurements. The pH at the end of the storage period was significantly lower than at the beginning. In vitro tests indicated that while the PC quality was acceptable upon expiration, it rapidly declined after expiration. In this setting where the vast majority of PCs were of good quality and within acceptable pH limits, daily, noninvasive routine pH measurement has limited added value in identifying quality-compromised PCs. © 2013 Sanquin Research. Transfusion © 2013 American Association of Blood Banks.
Wang, Bo; Liu, Anmin; Abdulla, Wael Al; Wang, Dianlong; Zhao, X S
2015-05-21
Electron transfer and lithium ion diffusion rates are the key factors limiting the lithium ion storage in anisotropic LiFePO4 electrodes. In this work, we employed a facile solvothermal method to synthesize a "platelet-on-sheet" LiFePO4/graphene composite (LFP@GNs), which is LiFePO4 nanoplatelets in situ grown on graphene sheets with highly oriented (010) facets of LiFePO4 crystals. Such a two-phase contact mode with graphene sheets cross-linked to form a three-dimensional porous network is favourable for both fast lithium ion and electron transports. As a result, the designed LFP@GNs displayed a high rate capability (∼56 mA h g(-1) at 60 C) and long life cycling stability (∼87% capacity retention over 1000 cycles at 10 C). For comparison purposes, samples ex situ modified with graphene (LFP/GNs) as well as pure LiFePO4 platelets (LFP) were also prepared and investigated. More importantly, the obtained LFP@GNs can be used as a basic unit for constructing more complex structures to further improve electrochemical performance, such as coating the exposed LFP surface with a thin layer of carbon to build a C@LFP@GN composite to further enhance its cycling stability (∼98% capacity retention over 1000 cycles at 10 C).
Bacterial growth kinetics in ACD-A apheresis platelets: comparison of plasma and PAS III storage.
Dumont, Larry J; Wood, Tammara A; Housman, Molly; Herschel, Louise; Brantigan, Barbara; Heber, Cheryl; Houghton, Jaime
2011-05-01
Our objective was to determine the growth kinetics of bacteria in leukoreduced apheresis platelets (LR-AP) in a platelet (PLT) additive solution (PAS; InterSol, Fenwal, Inc.) compared to LR-AP stored in plasma. Hyperconcentrated, double-dose LR-AP were collected from healthy donors with a separator (AMICUS, Fenwal, Inc.). LR-AP were evenly divided, InterSol was added to half (65% InterSol:35% plasma [PAS]), and PLTs in autologous plasma were used for a paired control (PL). Bacteria were inoculated into each LR-AP PAS/PL pair (0.5-1.6 colony-forming units [CFUs]/mL), and bacterial growth was followed for up to 7 days. Time to the end of the lag phase, doubling times, maximum concentration (conc-max), and time to maximum concentration (time-max) were estimated. Streptococcus viridans did not grow to detectable levels in either PAS or PL units. The other bacteria had no significant overall difference in the conc-max (p = 0.47) or time-max (p = 0.7) between PL and PAS LR-AP; PL had a 0.14 hours faster doubling rate (p = 0.023); and PAS had a 4.7 hours shorter lag time (p = 0.016). We observed that five index organisms will grow in LR-AP stored in a 35%:65% ratio of plasma to InterSol where initial bacterial concentrations are 0.5 to 1.6 CFUs/mL. The more rapid initiation of log-phase growth for bacteria within a PAS storage environment resulted in a bacterial concentration up to 4 logs higher in the PAS units compared to the plasma units at 24 hours, but with no difference in the conc-max. This may present an early bacterial detection advantage for PAS-stored PLTs. © 2010 American Association of Blood Banks.
Burnouf, Thierry; Chou, Ming-Li; Goubran, Hadi; Cognasse, Fabrice; Garraud, Olivier; Seghatchian, Jerard
2015-10-01
Blood cells and tissues generate heterogeneous populations of cell-derived vesicles, ranging from approximately 50 nm to 1 µm in diameter. Under normal physiological conditions and as an essential part of an energy-dependent natural process, microparticles (MPs) are continuously shed into the circulation from membranes of all viable cells such as megakaryocytes, platelets, red blood cells, white blood cells and endothelial cells. MP shedding can also be triggered by pathological activation of inflammatory processes and activation of coagulation or complement systems, or even by shear stress in the circulation. Structurally, MPs have a bilayered phospholipid structure exposing coagulant-active phosphatidylserine and expressing various membrane receptors, and they serve as cell-to-cell shuttles for bioactive molecules such as lipids, growth factors, microRNAs, and mitochondria. It was established that ex vivo processing of blood into its components, involving centrifugation, processing by various apheresis procedures, leucoreduction, pathogen reduction, and finally storage in different media and different types of blood bags, can impact MP generation and content. This is mostly due to exposure of the collected blood to anticoagulant/storage media and due to shear stresses or activation, contact with artificial surfaces, or exposure to various leucocyte-removal filters and pathogen-reduction treatments. Such artificially generated MPs, which are added to the original pool of MPs collected from the donor, may exhibit specific functional characteristics, as MPs are not an inert element of blood components. Not surprisingly, MPs' roles and functionality are therefore increasingly seen to be fully relevant to the field of transfusion medicine, and as a parameter of blood safety that must be considered in haemovigilance programmes. Continual advancements in assessment methods of MPs and storage lesions are gradually leading to a better understanding of the impacts of blood collection on MP generation, while clinical research should clarify links of MPs with transfusion reactions and certain clinical disorders. Harmonization and consensus in sampling protocols, sample handling and processing, and assessment methods are needed to achieve consensual interpretations. This review focuses on the role of MPs as an essential laboratory tool and as a most effective player in transfusion science and medicine and in health and disease. Copyright © 2015 Elsevier Ltd. All rights reserved.
NASA Astrophysics Data System (ADS)
Alvarez, Alejandro; Beche, Alexandre; Furano, Fabrizio; Hellmich, Martin; Keeble, Oliver; Rocha, Ricardo
2012-12-01
The Disk Pool Manager (DPM) is a lightweight solution for grid enabled disk storage management. Operated at more than 240 sites it has the widest distribution of all grid storage solutions in the WLCG infrastructure. It provides an easy way to manage and configure disk pools, and exposes multiple interfaces for data access (rfio, xroot, nfs, gridftp and http/dav) and control (srm). During the last year we have been working on providing stable, high performant data access to our storage system using standard protocols, while extending the storage management functionality and adapting both configuration and deployment procedures to reuse commonly used building blocks. In this contribution we cover in detail the extensive evaluation we have performed of our new HTTP/WebDAV and NFS 4.1 frontends, in terms of functionality and performance. We summarize the issues we faced and the solutions we developed to turn them into valid alternatives to the existing grid protocols - namely the additional work required to provide multi-stream transfers for high performance wide area access, support for third party copies, credential delegation or the required changes in the experiment and fabric management frameworks and tools. We describe new functionality that has been added to ease system administration, such as different filesystem weights and a faster disk drain, and new configuration and monitoring solutions based on the industry standards Puppet and Nagios. Finally, we explain some of the internal changes we had to do in the DPM architecture to better handle the additional load from the analysis use cases.
Johnson, Lacey; Coorey, Craig P; Marks, Denese C
2014-08-01
Cryopreservation of platelets (PLTs) at -80°C with dimethyl sulfoxide (DMSO) can extend the shelf life from 5 days to 2 years. Cryopreserved PLTs are reported to have a greater in vivo hemostatic effect than liquid-stored PLTs. As such, the aim of this study was to understand the mechanisms responsible for the hemostatic potential of cryopreserved PLTs and the contribution of the reconstitution solution to this activity. DMSO (5% final concentration) was added to buffy coat-derived PLTs, followed by prefreeze removal of DMSO and storage at -80°C. Cryopreserved PLTs (n=8 per group) were thawed at 37°C, reconstituted with either 1 unit of thawed frozen plasma or PLT additive solution (PAS-G). In vitro assays were performed before freezing and after thawing to assess the hemostatic activity of PLTs. Cryopreserved PLTs expressed high levels of phosphatidylserine and contained significantly more phosphatidylserine-positive PLT microparticles than liquid-stored PLTs. This was accompanied by a significant decrease in the time to clot formation and clot strength, as measured by thromboelastography. The supernatant from cryopreserved PLTs was sufficient to reduce the phosphatidylserine-dependent clotting time and increase the thrombin generation potential. Overall, plasma-reconstituted cryopreserved PLTs were more procoagulant than those reconstituted in PAS-G. PLT cryopreservation results in the generation of phosphatidylserine-expressing PLT microparticles which contribute to the hemostatic activity. Understanding the hemostatic activity of these components may assist in extending the use of these specialized components beyond military applications. © 2014 Australian Red Cross Blood Service. Transfusion © 2014 AABB.
Carbon Uptake and Storage in Old-Growth and Second-Growth Forests in Central Vermont
NASA Astrophysics Data System (ADS)
Lloyd, A. H.; Weisser, O.
2013-12-01
Managing forests towards the goal of maximizing carbon uptake and storage provides an important tool for climate change mitigation. There is significant spatial and temporal variation among forests, even within an ecosystem type, in annual uptake and storage of carbon. Understanding the causes for that variation is important in refining management practices and restoration goals that promote carbon storage. We explore the variation in carbon storage and uptake among forests differing in age in central Vermont, comparing young, intermediate-aged, and old-growth forests. We generally expected that younger forests would have a higher annual uptake of carbon than older forests. Significant uncertainty exists, however, about the temporal trajectory from a young, rapidly growing forest to an old-growth forest that may be in a steady-state, with no net uptake of carbon. Within each forest, we compare differences among functional groups of species (e.g., hardwoods versus softwoods) in contribution to overall forest carbon uptake and storage. Our study sites include an old-growth hemlock/mixed hardwood forest that has not been directly affected by human activities, and which contains trees upwards of 350 years old; a 130-year-old mixed hardwood forest that has recolonized former pasture land; and a 90-year-old mixed hardwood forest on formerly agricultural floodplain land. Carbon storage in live and dead biomass pools was estimated from allometric equations, based on repeated measurements of tree diameters in permanently marked study plots. Historical patterns of carbon storage in living biomass were estimated by reconstructing tree diameter from measured increment cores, and then estimating the living biomass in each year. As expected, the old-growth forest stored almost twice the C in live biomass as the two second-growth forests, which stored equivalent amounts of carbon, despite the difference in age. Dead biomass was a larger pool of C in the old-growth forest than in the two second-growth forests, but still contained only a quarter of the C of the live biomass pool. Both repeated measurements of tree diameters and tree-ring reconstructions of historical patterns of C accumulation suggested that all three forests were continuing to accumulate C in biomass, but the rate differed substantially among sites, with the lowest rates of accumulation occurring in the old-growth forest. Within the old-growth forest, the fastest rates of biomass accumulation occurred in younger hardwoods, which appear to have colonized old canopy gaps in the mid-1800s. Together, these results are consistent with prior research suggesting that C continues to accumulate in temperate forests for hundreds of years. Both species differences and forest age, however, have a significant effect on C uptake and storage.
The legacy of forest harvest and burning on ecosystem carbon storage in the northern midwest, USA
NASA Astrophysics Data System (ADS)
Gough, C. M.; Vogel, C. S.; Harrold, K. H.; George, K. D.; Curtis, P. S.
2005-12-01
Over 90 % of the forested area in the upper Great Lakes region was harvested by the early 20th century. In many cases, harvests were followed by uncontrolled burns, similar to current patterns of disturbance in many developing countries. While afforestation in the northern midwest has resulted in increased regional carbon (C) storage, the rate of C storage by forests will depend on the severity of prior disturbance and consequent changes in site quality. We were interested in how long the legacy of poor management practices from the early 20th century would be reflected in forest C storage rates. We investigated C cycling and storage following disturbance in mixed deciduous forests of northern lower Michigan, USA. Study plots ranged in age from 6 to 68 yrs and were created following experimental clear-cut harvesting and fire disturbance. Annual C storage was estimated biometrically from measurements of wood, leaf, fine root, and woody debris mass, mass losses to herbivory, soil carbon content, and soil respiration. Maximum annual carbon storage, or net ecosystem production (NEP), in the disturbed stands was 50 % lower than that of adjacent, undisturbed forest. This decrease was caused by a reduction in site quality following disturbance. However, during regrowth the cut and burned forest rapidly became a net C sink, storing 0.86 Mg C ha-1 yr-1 after six yrs. Carbon storage reached a peak of 1.00 Mg C ha-1 yr-1 after 50 yrs and declined to 0.57 Mg C ha-1 yr-1 after 68 yrs. Above- and below-ground net primary production (NPP) averaged 42 and 59 % of total NPP, respectively, with fine root litter production accounting for 57 % of total NPP. Soil heterotrophic respiration was high, ranging from 4.55 Mg C ha-1 yr-1 in the 6-yr-old stand to 5.74 Mg C ha-1 yr-1 in the 50-yr-old stand. Soil C and coarse woody debris pools exhibited a U-shaped trend over time following disturbance. Mineral soil and coarse woody debris pools lost C at a combined annual rate of 1.10 Mg C ha-1 yr-1 in the 6-yr-old stand, but these pools accrued C at a rate of 0.30 Mg C ha--1 yr-1 in the 68-yr-old stand. Detritus inputs augmented soil C six years after harvest and this legacy C persisted in the oldest, 68-yr-old stand. This resulted in higher soil C than in an adjacent undisturbed mature forest. These results demonstrate that lasting decreases in site quality following disturbance result in long-term reductions in forest C storage.
LPT. Plot plan and site layout. Includes shield test pool/EBOR ...
LPT. Plot plan and site layout. Includes shield test pool/EBOR facility. (TAN-645 and -646) low power test building (TAN-640 and -641), water storage tanks, guard house (TAN-642), pump house (TAN-644), driveways, well, chlorination building (TAN-643), septic system. Ralph M. Parsons 1229-12 ANP/GE-7-102. November 1956. Approved by INEEL Classification Office for public release. INEEL index code no. 038-0102-00-693-107261 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID
NASA Astrophysics Data System (ADS)
Lopez, Juan Manuel
Layer-by-layer self-assembly (LbL) is a technique that generates engineered nano-scale films, coatings, and particles. These nanoscale films have recently been used in multiple biomedical applications. Concurrently, microfabrication methods and advances in microfluidics are being developed and combined to create "Lab-on-a-Chip" technologies. The potential to perform complex biological assays in vitro as a first-line screening technique before moving on to animal models has made the concept of lab on a chip a valuable research tool. Prior studies in the Biofluids Laboratory at Louisiana Tech have used layer-by-layer and in vitro biological assays to study thrombogenesis in a controlled, repeatable, engineered environment. The reliability of these previously established techniques was unsatisfactory for more complex cases such as chemical and shear stress interactions. The work presented in this dissertation was performed to test the principal assumptions behind the established laboratory methodologies, suggest improvements where needed, and test the impact of these improvements on accuracy and repeatability. The assumptions to be tested were: (1) The fluorescence microscopy (FM) images of acridine orange-tagged platelets accurately provide a measure of percent area of surface covered by platelets; (2) fibrinogen coatings can be accurately controlled, interact with platelets, and do not interfere with the ability to quantify platelet adhesion; and (3) the dependence of platelet adhesion on chemical agents, as measured with the modified methods, generally agrees with results obtained from our previous methods and with known responses of platelets that have been documented in the literature. The distribution of fibrinogen on the final LbL surface generated with the standard, static process (s-LbL) was imaged by tagging the fibrinogen with an anti-fibrinogen antibody bound to fluorescein isothiocyanate (FITC). FITC FM images and acridine orange FM images were taken sequentially at selected surface locations to generate a composite overlap of presumed platelet adhesion as a function of fibrinogen distribution. The method was unable to distinguish the surface from the adhered cells. The surface inhomogeneity and porosity retained a large amount of acridine orange stain, even in the absence of platelets, and components in the platelet-rich plasma (PRP) were found to fix acridine orange in a mode that fluoresced in the FITC imaging FM. Both of these problems obfuscated the platelet adhesion FM results when using s-LbL surfaces and acridine orange staining of platelets. A dynamic process (d-LbL) was developed in which a solution of the molecule to be layered was constantly washed over the surface, and was constantly mixed to maintain a more homogeneous distribution of solute relative to the surface during the layering process. The d-LbL surfaces were tested as described above, and found to reduce the size and number of regions of anomalous acridine orange pooling trapped by the surface, providing a greater consistency and reliability in identifying platelets. The improved surface was then used in a series of platelet adhesion experiments under static and dynamic flow conditions, and with and without the chemical additive L-arginine. The complex microcharmel system used in prior studies was replaced with a simpler system involving fewer nuisance variables for these tests. The tests were performed on both collagen and fibrinogen surfaces. Collagen has been used as a thrombogenic surface in multiple studies in the literature, but produces additional variables in thrombogenesis control that are avoided when fibrinogen is used. In these tests, fibrinogen was found to be as thrombogenic as collagen, and platelet coverage of both biointerfaces was reduced by L-arginine in a manner similar to previously reported work. The simpler system differed from the previous microchannel system in important factors: (1) It exposed the platelets to much lower shear stresses; (2) It introduced an oscillatory flow, which introduced a higher degree of variability in the adhesion than previously reported; (3) the previous work had not removed the acridine orange surface problems. Therefore, a direct comparison between results was not possible. The new d-LbL surface process was successful in testing the basic assumptions. Testing showed that the new process eliminated the anomalous acridine orange retention problem during fluorescence imaging. This improvement in fluorescence response meant that the FM results matched the platelet adhesion on plain glass slides and adhesion reported by others in microfluidic flows. The chemical additive responses behaved as expected, with an increase in L-arginine contributing to a decrease in thrombogenesis under dynamic conditions, but not under static conditions.
Lye, David C; Archuleta, Sophia; Syed-Omar, Sharifah F; Low, Jenny G; Oh, Helen M; Wei, Yuan; Fisher, Dale; Ponnampalavanar, Sasheela S L; Wijaya, Limin; Lee, Linda K; Ooi, Eng-Eong; Kamarulzaman, Adeeba; Lum, Lucy C; Tambyah, Paul A; Leo, Yee-Sin
2017-04-22
Dengue is the commonest vector-borne infection worldwide. It is often associated with thrombocytopenia, and prophylactic platelet transfusion is widely used despite the dearth of robust evidence. We aimed to assess the efficacy and safety of prophylactic platelet transfusion in the prevention of bleeding in adults with dengue and thrombocytopenia. We did an open-label, randomised, superiority trial in five hospitals in Singapore and Malaysia. We recruited patients aged at least 21 years who had laboratory-confirmed dengue (confirmed or probable) and thrombocytopenia (≤20 000 platelets per μL), without persistent mild bleeding or any severe bleeding. Patients were assigned (1:1), with randomly permuted block sizes of four or six and stratified by centre, to receive prophylactic platelet transfusion in addition to supportive care (transfusion group) or supportive care alone (control group). In the transfusion group, 4 units of pooled platelets were given each day when platelet count was 20 000 per μL or lower; supportive care consisted of bed rest, fluid therapy, and fever and pain medications. The primary endpoint was clinical bleeding (excluding petechiae) by study day 7 or hospital discharge (whichever was earlier), analysed by intention to treat. Safety outcomes were analysed according to the actual treatment received. This study was registered with ClinicalTrials.gov, number NCT01030211, and is completed. Between April 29, 2010, and Dec 9, 2014, we randomly assigned 372 patients to the transfusion group (n=188) or the control group (n=184). The intention-to-treat analysis included 187 patients in the transfusion group (one patient was withdrawn immediately) and 182 in the control group (one was withdrawn immediately and one did not have confirmed or probable dengue). Clinical bleeding by day 7 or hospital discharge occurred in 40 (21%) patients in the transfusion group and 48 (26%) patients in the control group (risk difference -4·98% [95% CI -15·08 to 5·34]; relative risk 0·81 [95% CI 0·56 to 1·17]; p=0·16). 13 adverse events occurred in the transfusion group and two occurred in the control group (5·81% [-4·42 to 16·01]; 6·26 [1·43 to 27·34]; p=0·0064). Adverse events that were possibly, probably, or definitely related to transfusion included three cases of urticaria, one maculopapular rash, one pruritus, and one chest pain, as well as one case each of anaphylaxis, transfusion-related acute lung injury, and fluid overload that resulted in serious adverse events. No death was reported. In adult patients with dengue and thrombocytopenia, prophylactic platelet transfusion was not superior to supportive care in preventing bleeding, and might be associated with adverse events. National Medical Research Council, Singapore. Copyright © 2017 Elsevier Ltd. All rights reserved.
Lu, Xuyang; Yan, Yan; Sun, Jian; Zhang, Xiaoke; Chen, Youchao; Wang, Xiaodan; Cheng, Genwei
2015-10-01
In recent decades, alpine grasslands have been seriously degraded on the Tibetan Plateau and grazing exclusion by fencing has been widely adopted to restore degraded grasslands since 2004. To elucidate how alpine grasslands carbon (C), nitrogen (N), and phosphorus (P) storage responds to this management strategy, three types of alpine grassland in nine counties in Tibet were selected to investigate C, N, and P storage in the environment by comparing free grazing (FG) and grazing exclusion (GE) treatments, which had run for 6-8 years. The results revealed that there were no significant differences in total ecosystem C, N, and P storage, as well as the C, N, and P stored in both total biomass and soil (0-30 cm) fractions between FG and GE grasslands. However, precipitation played a key role in controlling C, N, and P storage and distribution. With grazing exclusion, C and N stored in aboveground biomass significantly increased by 5.7 g m(-2) and 0.1 g m(-2), respectively, whereas the C and P stored in the soil surface layer (0-15 cm) significantly decreased by 862.9 g m(-2) and 13.6 g m(-2), respectively. Furthermore, the storage of the aboveground biomass C, N, and P was positively correlated with vegetation cover and negatively correlated with the biodiversity index, including Pielou evenness index, Shannon-Wiener diversity index, and Simpson dominance index. The storage of soil surface layer C, N, and P was positively correlated with soil silt content and negatively correlated with soil sand content. Our results demonstrated that grazing exclusion had no impact on total C, N, and P storage, as well as C, N, and P in both total biomass and soil (0-30 cm) fractions in the alpine grassland ecosystem. However, grazing exclusion could result in increased aboveground biomass C and N pools and decreased soil surface layer (0-15 cm) C and P pools.
NASA Astrophysics Data System (ADS)
Dye, A.; Alexander, M. R.; Bishop, D.; Pederson, N.; Hessl, A. E.
2016-12-01
Storage of carbon in terrestrial plants and soils directly reduces atmospheric carbon concentration, and it is thereby imperative to improve our understanding of where carbon is being stored and released in an ecosystem and how storages and releases are changing over time. At data-rich sites, coupling alternative measurements of carbon flux can improve this understanding. Here, we present a methodology to inversely model stand-level net storage and release of above- and belowground carbon over a period of 1-2 decades using co-located tree-ring plots and eddy covariance towers at three eastern U.S. forests. We reconstructed annual aboveground wood production (aNPP) from tree rings collected near eddy covariance towers. We compared our aNPP reconstructions with annual tower NEE to address whether interannual variations are correlated. Despite modest correlation, we observed magnitude differences between both records that vary annually. We interpret these differences as indicative of changes in belowground carbon storage, i.e. an aNPP:NEE ratio > 1 indicates a net release of belowground carbon and a ratio < 1 a net storage of belowground carbon. For this interpretation, we assume the following: a) carbon not directed to above or belowground pools is insignificant, b) carbon not stored above ground is stored below ground, and c) mature trees do not add to a storage pool at a higher level every year. While the offset between biometric aNPP and tower NEE could partially be attributed to the diversion of assimilated carbon to nonstructural carbohydrates instead of growth, we argue that this becomes a less important factor over longer time scales in a mature tree. Our approach does not quantify belowground NPP or allocation, but we present a method for estimating belowground carbon storage and release at the stand level, an otherwise difficult task at this scale due to heterogeneity across the stand.
Charan, Jaykaran; Saxena, Deepak; Goyal, Jagdish Prasad; Yasobant, Sandul
2016-01-01
Dengue is an infectious disease associated with high mortality and morbidity. Being a viral disease, there is no specific drug available for treatment. There are some reports that Carica papaya leaf extract may improve the clinical condition of dengue patients; however, to support this, at present, there is no systematically searched and synthesized evidence available. This systematic review and meta-analysis was designed to search the available evidence related to the efficacy and safety of C. papaya leaf extract in dengue and to synthesize the evidence in meaningful form through meta-analysis so that inference can be drawn. Randomized controlled trials related to the efficacy and safety of C. papaya leaf extract in dengue were searched from PubMed, Cochrane Clinical Trial Registry and Google Scholar. The primary endpoint was mortality, and secondary endpoints were increase in platelet count, hospitalization days, and Grade 3 and 4 adverse events. Data related to primary and secondary endpoints were pooled together and analyzed by review manager (Review Manager (RevMan) Copenhagen: The Nordic Cochrane Centre, The Cochrane Collaboration, Denmark) software. The random effect model was used. The bias was analyzed by the Cochrane risk of bias tool. Total four trials enrolling 439 subjects were included in the analysis. Of 439 subjects, data of 377 subjects were available for analysis. C. papaya leaf extract was found to be associated with increase in platelet count in the overall analysis (mean difference [MD] =20.27 [95% confidence interval (CI) 6.21-34.73; P = 0.005]) and analysis after 4 th day (MD = 28.25 [95% CI 14.14-42.37; P < 0.0001]). After 48 h, there was no significant difference between C. papaya and control group (MD = 13.38 [95% CI - 7.71-34.51; P = 0.21]). There was significant decrease in hospitalization days in the C. papaya group (MD = 1.90 [95% CI 1.62-2.18; P < 0.00001]). Because of nonavailability of data in published clinical trials, mortality, and adverse events cannot be pooled. C. papaya leaf extract can be considered as a potential candidate for increase in platelet count in patients of dengue, however; there is need of high-quality evidence in the form of large clinical trials before a decision related to the use of such extract is made.
DPM — efficient storage in diverse environments
NASA Astrophysics Data System (ADS)
Hellmich, Martin; Furano, Fabrizio; Smith, David; Brito da Rocha, Ricardo; Álvarez Ayllón, Alejandro; Manzi, Andrea; Keeble, Oliver; Calvet, Ivan; Regala, Miguel Antonio
2014-06-01
Recent developments, including low power devices, cluster file systems and cloud storage, represent an explosion in the possibilities for deploying and managing grid storage. In this paper we present how different technologies can be leveraged to build a storage service with differing cost, power, performance, scalability and reliability profiles, using the popular storage solution Disk Pool Manager (DPM/dmlite) as the enabling technology. The storage manager DPM is designed for these new environments, allowing users to scale up and down as they need it, and optimizing their computing centers energy efficiency and costs. DPM runs on high-performance machines, profiting from multi-core and multi-CPU setups. It supports separating the database from the metadata server, the head node, largely reducing its hard disk requirements. Since version 1.8.6, DPM is released in EPEL and Fedora, simplifying distribution and maintenance, but also supporting the ARM architecture beside i386 and x86_64, allowing it to run the smallest low-power machines such as the Raspberry Pi or the CuBox. This usage is facilitated by the possibility to scale horizontally using a main database and a distributed memcached-powered namespace cache. Additionally, DPM supports a variety of storage pools in the backend, most importantly HDFS, S3-enabled storage, and cluster file systems, allowing users to fit their DPM installation exactly to their needs. In this paper, we investigate the power-efficiency and total cost of ownership of various DPM configurations. We develop metrics to evaluate the expected performance of a setup both in terms of namespace and disk access considering the overall cost including equipment, power consumptions, or data/storage fees. The setups tested range from the lowest scale using Raspberry Pis with only 700MHz single cores and a 100Mbps network connections, over conventional multi-core servers to typical virtual machine instances in cloud settings. We evaluate the combinations of different name server setups, for example load-balanced clusters, with different storage setups, from using a classic local configuration to private and public clouds.
Waste Encapsulation and Storage Facility (WESF) Dangerous Waste Training Plan (DWTP)
DOE Office of Scientific and Technical Information (OSTI.GOV)
SIMMONS, F.M.
2000-03-29
This Waste Encapsulation Storage Facility (WESF) Dangerous Waste Training Plan (DWTP) applies to personnel who perform work at, or in support of WESF. The plan, along with the names of personnel, may be given to a regulatory agency inspector upon request. General workers, subcontractors, or visiting personnel who have not been trained in the management of dangerous wastes must be accompanied by an individual who meets the requirements of this training plan. Dangerous waste management includes handling, treatment, storage, and/or disposal of dangerous and/or mixed waste. Dangerous waste management units covered by this plan include: less-than-90-day accumulation area(s); pool cellsmore » 1-8 and 12 storage units; and process cells A-G storage units. This training plan describes general requirements, worker categories, and provides course descriptions for operation of the WESF permitted miscellaneous storage units and the Less-than-90-Day Accumulation Areas.« less
How well do we succeed in modeling the global soil carbon pools?
NASA Astrophysics Data System (ADS)
Viskari, T.; Liski, J.
2017-12-01
Terrestrial carbon pools are a crucial part of the global carbon cycle. Carbon from vegetation is deposited to the soil, which in turn releases carbon dioxide back to the atmosphere through heterotrophic respiration. The resulting soil carbon storage in the largest on land. While there are continuous efforts to improve the modeling of global soil carbon and how this storage is affected by climate change, this research requires still a more reliable baseline on how well the models estimate the current global soil carbon pools. Especially such comparisons are important for identifying the major challenges in the current soil carbon models. Here, we used the Yasso soil carbon model to create a global soil carbon map at a 0.5 degree resolution based on the available climate, land cover and vegetation productivity information. Yasso model describes the soil carbon cycling by pools that represent the breaking down of dead organic matter. We compared the model results to a measurement based projection of global soil carbon pools, and we examined the differences and spatial correlations between the two maps. In our findings, the modelled predictions captured the overall soil carbon distributions within 5 kgCm-2 on 63 % of the land area. The spatial distributions fit each other as well. The average soil carbon is smaller with the Yasso prediction ( 8.5 kg m-2) than with the measurement map ( 10 kg m-2) and there are notable areas, such as Siberia and Southern North America, where there are large differences between the model predictions and measurements. These results not only encourage future development of soil carbon models, but also highlight problem areas to focus and improve upon.
Laboratory investigation and simulation of breakthrough curves in karst conduits with pools
NASA Astrophysics Data System (ADS)
Zhao, Xiaoer; Chang, Yong; Wu, Jichun; Peng, Fu
2017-12-01
A series of laboratory experiments are performed under various hydrological conditions to analyze the effect of pools in pipes on breakthrough curves (BTCs). The BTCs are generated after instantaneous injections of NaCl tracer solution. In order to test the feasibility of reproducing the BTCs and obtain transport parameters, three modeling approaches have been applied: the equilibrium model, the linear graphical method and the two-region nonequilibrium model. The investigation results show that pools induce tailing of the BTCs, and the shapes of BTCs depend on pool geometries and hydrological conditions. The simulations reveal that the two-region nonequilibrium model yields the best fits to experimental BTCs because the model can describe the transient storage in pools by the partition coefficient and the mass transfer coefficient. The model parameters indicate that pools produce high dispersion. The increased tailing occurs mainly because the partition coefficient decreases, as the number of pools increases. When comparing the tracer BTCs obtained using the two types of pools with the same size, the more appreciable BTC tails that occur for symmetrical pools likely result mainly from the less intense exchange between the water in the pools and the water in the pipe, because the partition coefficients for the two types of pools are virtually identical. Dispersivity values decrease as flow rates increase; however, the trend in dispersion is not clear. The reduced tailing is attributed to a decrease in immobile water with increasing flow rate. It provides evidence for hydrodynamically controlled tailing effects.
Enhanced Shear-induced Platelet Aggregation Due to Low-temperature Storage
2013-07-01
conjugated sheep polyclonal anti-VWF antibody , goat polyclonal anti-mouse FITC and PE secondary antibodies , and immunoglobulin G isotype control antibodies ...of fresh or stored PRP was incubated with 25 mg/mL FITC- conjugated AK2 or unconjugated SZ2 or VM16d antibodies for 20 minutes. In case of SZ2 and...VM16d, the primary antibody -bound PLTs were incubated with a PE- conjugated secondary antibody . The antibody - bound PLTs were then fixed in 400 mL of 1
Entropy, pumped-storage and energy system finance
NASA Astrophysics Data System (ADS)
Karakatsanis, Georgios
2015-04-01
Pumped-storage holds a key role for integrating renewable energy units with non-renewable fuel plants into large-scale energy systems of electricity output. An emerging issue is the development of financial engineering models with physical basis to systematically fund energy system efficiency improvements across its operation. A fundamental physically-based economic concept is the Scarcity Rent; which concerns the pricing of a natural resource's scarcity. Specifically, the scarcity rent comprises a fraction of a depleting resource's full price and accumulates to fund its more efficient future use. In an integrated energy system, scarcity rents derive from various resources and can be deposited to a pooled fund to finance the energy system's overall efficiency increase; allowing it to benefit from economies of scale. With pumped-storage incorporated to the system, water upgrades to a hub resource, in which the scarcity rents of all connected energy sources are denominated to. However, as available water for electricity generation or storage is also limited, a scarcity rent upon it is also imposed. It is suggested that scarcity rent generation is reducible to three (3) main factors, incorporating uncertainty: (1) water's natural renewability, (2) the energy system's intermittent components and (3) base-load prediction deviations from actual loads. For that purpose, the concept of entropy is used in order to measure the energy system's overall uncertainty; hence pumped-storage intensity requirements and generated water scarcity rents. Keywords: pumped-storage, integration, energy systems, financial engineering, physical basis, Scarcity Rent, pooled fund, economies of scale, hub resource, uncertainty, entropy Acknowledgement: This research was funded by the Greek General Secretariat for Research and Technology through the research project Combined REnewable Systems for Sustainable ENergy DevelOpment (CRESSENDO; grant number 5145)
Collection, Storage, and Preparation of Human Blood Cells
Dagur, Pradeep K.; McCoy, J. Philip
2015-01-01
Human peripheral blood is often studied by flow cytometry in both the research and clinical laboratories. The methods for collection, storage, and preparation of peripheral blood will vary depending on the cell lineage to be examined as well as the type of assay to be performed. This unit presents protocols for collection of blood, separation of leukocytes from whole blood by lysis of erythrocytes, isolating mononuclear cells by density gradient separation, and assorted non-flow sorting methods, such as magnetic bead separations, for enriching specific cell populations, including monocytes, T lymphocytes, B lymphocytes, neutrophils,, , and platelets prior to flow cytometric analysis. A protocol is also offered for cryopreservation of cells since clinical research often involves retrospective flow cytometric analysis of samples stored over a period of months or years. PMID:26132177
Tang, Hai Ming; Cheng, Kai Kai; Xiao, Xiao Ping; Tang, Wen Guang; Wang, Ke; Li, Chao; Zhang, Fan; Sun, Yu Tao
2017-02-01
In a double cropping rice field experiment, effects of five winter cover crops on the total organic carbon (TOC), active organic carbon (AOC), carbon pool management index (CPMI) and organic carbon storage were studied in three soil layers (0-5, 5-10 and 10-20 cm).Winter cover crops of ryegrass (Ry), Chinese milk vetch (Mv), potato (Po), and rape (Ra) between two rice crops were compared with fallow as control (CK). The results showed that the TOC and AOC contents under Ry, Mv, Po and Ra treatments were higher than those of CK in all three la-yers. Meanwhile, the TOC and AOC contents in Po treatment were higher than those of other treatments. Compared with CK, the AOC, activity index (AI), carbon pool index (CPI) and CPMI in the soil were improved through the recycling of winter cover crops straw. The AOC, AI, CPI and CPMI in the studied layers increased in order of Po>Mv>Ry>Ra>CK. The results indicated that the recycling of winter cover crops straw promoted the storage of SOC in the 0-20 cm soil profile as compared with CK. The strongest effect of the winter cover crops on the SOC storage occurred in Mv treatment, followed by Mv and Po treatments, and the SOC storage increased with the increasing soil depth.
The Fate of Nitrate in Intertidal Permeable Sediments
Marchant, Hannah K.; Lavik, Gaute; Holtappels, Moritz; Kuypers, Marcel M. M.
2014-01-01
Coastal zones act as a sink for riverine and atmospheric nitrogen inputs and thereby buffer the open ocean from the effects of anthropogenic activity. Recently, microbial activity in sandy permeable sediments has been identified as a dominant source of N-loss in coastal zones, namely through denitrification. Some of the highest coastal denitrification rates measured so far occur within the intertidal permeable sediments of the eutrophied Wadden Sea. Still, denitrification alone can often account for only half of the substantial nitrate (NO3 −) consumption. Therefore, to investigate alternative NO3 − sinks such as dissimilatory nitrate reduction to ammonium (DNRA), intracellular nitrate storage by eukaryotes and isotope equilibration effects we carried out 15NO3 − amendment experiments. By considering all of these sinks in combination, we could quantify the fate of the 15NO3 − added to the sediment. Denitrification was the dominant nitrate sink (50–75%), while DNRA, which recycles N to the environment accounted for 10–20% of NO3 − consumption. Intriguingly, we also observed that between 20 and 40% of 15NO3 − added to the incubations entered an intracellular pool of NO3 − and was subsequently respired when nitrate became limiting. Eukaryotes were responsible for a large proportion of intracellular nitrate storage, and it could be shown through inhibition experiments that at least a third of the stored nitrate was subsequently also respired by eukaryotes. The environmental significance of the intracellular nitrate pool was confirmed by in situ measurements which revealed that intracellular storage can accumulate nitrate at concentrations six fold higher than the surrounding porewater. This intracellular pool is so far not considered when modeling N-loss from intertidal permeable sediments; however it can act as a reservoir for nitrate during low tide. Consequently, nitrate respiration supported by intracellular nitrate storage can add an additional 20% to previous nitrate reduction estimates in intertidal sediments, further increasing their contribution to N-loss. PMID:25127459
The fate of nitrate in intertidal permeable sediments.
Marchant, Hannah K; Lavik, Gaute; Holtappels, Moritz; Kuypers, Marcel M M
2014-01-01
Coastal zones act as a sink for riverine and atmospheric nitrogen inputs and thereby buffer the open ocean from the effects of anthropogenic activity. Recently, microbial activity in sandy permeable sediments has been identified as a dominant source of N-loss in coastal zones, namely through denitrification. Some of the highest coastal denitrification rates measured so far occur within the intertidal permeable sediments of the eutrophied Wadden Sea. Still, denitrification alone can often account for only half of the substantial nitrate (NO3-) consumption. Therefore, to investigate alternative NO3- sinks such as dissimilatory nitrate reduction to ammonium (DNRA), intracellular nitrate storage by eukaryotes and isotope equilibration effects we carried out 15NO3- amendment experiments. By considering all of these sinks in combination, we could quantify the fate of the 15NO3- added to the sediment. Denitrification was the dominant nitrate sink (50-75%), while DNRA, which recycles N to the environment accounted for 10-20% of NO3- consumption. Intriguingly, we also observed that between 20 and 40% of 15NO3- added to the incubations entered an intracellular pool of NO3- and was subsequently respired when nitrate became limiting. Eukaryotes were responsible for a large proportion of intracellular nitrate storage, and it could be shown through inhibition experiments that at least a third of the stored nitrate was subsequently also respired by eukaryotes. The environmental significance of the intracellular nitrate pool was confirmed by in situ measurements which revealed that intracellular storage can accumulate nitrate at concentrations six fold higher than the surrounding porewater. This intracellular pool is so far not considered when modeling N-loss from intertidal permeable sediments; however it can act as a reservoir for nitrate during low tide. Consequently, nitrate respiration supported by intracellular nitrate storage can add an additional 20% to previous nitrate reduction estimates in intertidal sediments, further increasing their contribution to N-loss.
Yang, Meng; Li, Yong Fu; Li, Yong Chun; Xiao, Yong Heng; Yue, Tian; Jiang, Pei Kun; Zhou, Guo Mo; Liu, Juan
2016-11-18
In order to elucidate the effects of intensive management on soil carbon pool, nitrogen pool, enzyme activities in Moso bamboo (Phyllostachys pubescens) plantations, we collected soil samples from the soil surface (0-20 cm) and subsurface (20-40 cm) layers in the adjacent Moso bamboo plantations with extensive and intensive managements in Sankou Township, Lin'an City, Zhejiang Province. We determined different forms of C, N and soil invertase, urease, catalase and acid phosphatase activities. The results showed that long-term intensive management of Moso bamboo plantations significantly decreased the content and storage of soil organic carbon (SOC), with the SOC storage in the soil surface and subsurface layers decreased by 13.2% and 18.0%, respectively. After 15 years' intensive management of Masoo bamboo plantations, the contents of soil water soluble carbon (WSOC), hot water soluble carbon (HWSOC), microbial carbon (MBC) and readily oxidizable carbon (ROC) were significantly decreased in the soil surface and subsurface layers. The soil N storage in the soil surface and subsurface layers in intensively managed Moso bamboo plantations increased by 50.8% and 36.6%, respectively. Intensive management significantly increased the contents of nitrate-N (NO 3 - -N) and ammonium-N (NH 4 + -N), but decreased the contents of water-soluble nitrogen (WSON) and microbial biomass nitrogen (MBN). After 15 years' intensive management of Masoo bamboo plantations, the soil invertase, urease, catalase and acid phosphatase activities in the soil surface layer were significantly decreased, the soil acid phosphatase activity in the soil subsurface layer were significantly decreased, and other enzyme activities in the soil subsurface layer did not change. In conclusion, long-term intensive management led to a significant decline of soil organic carbon storage, soil labile carbon and microbial activity in Moso bamboo plantations. Therefore, we should consider the use of organic fertilizer in the intensive mana-gement process for the sustainable management of Moso bamboo plantations in the future.
Havekes, Robbert; Canton, David A.; Park, Alan J.; Huang, Ted; Nie, Ting; Day, Jonathan P.; Guercio, Leonardo A.; Grimes, Quinn; Luczak, Vincent; Gelman, Irwin H.; Baillie, George S.; Scott, John D.; Abel, Ted
2012-01-01
A kinase-anchoring proteins (AKAPs) organize compartmentalized pools of Protein Kinase A (PKA) to enable localized signaling events within neurons. However, it is unclear which of the many expressed AKAPs in neurons target PKA to signaling complexes important for long-lasting forms of synaptic plasticity and memory storage. In the forebrain, the anchoring protein gravin recruits a signaling complex containing PKA, PKC, calmodulin, and PDE4D to the β2-adrenergic receptor. Here, we show that mice lacking the α-isoform of gravin have deficits in PKA-dependent long-lasting forms of hippocampal synaptic plasticity including β2-adrenergic receptor-mediated plasticity, and selective impairments of long-term memory storage. Further, both hippocampal β2-adrenergic receptor phosphorylation by PKA, and learning-induced activation of ERK, are attenuated in the CA1 region of the hippocampus in mice lacking gravin-α. We conclude that gravin compartmentalizes a significant pool of PKA that regulates learning-induced β2-adrenergic receptor signaling and ERK activation in the hippocampus in vivo, organizing molecular interactions between glutamatergic and noradrenergic signaling pathways for long-lasting synaptic plasticity, and memory storage. PMID:23238728
FRAPCON analysis of cladding performance during dry storage operations
DOE Office of Scientific and Technical Information (OSTI.GOV)
Richmond, David J.; Geelhood, Kenneth J.
There is an increasing need in the U.S. and around the world to move used nuclear fuel from wet storage in fuel pools to dry storage in casks stored at independent spent fuel storage installations (ISFSI) or interim storage sites. The NRC limits cladding temperature to 400°C while maintaining cladding hoop stress below 90 MPa in an effort to avoid radial hydride reorientation. An analysis was conducted with FRAPCON-4.0 on three modern fuel designs with three representative used nuclear fuel storage temperature profiles that peaked at 400 °C. Results were representative of the majority of U.S. LWR fuel. They conservativelymore » showed that hoop stress remains below 90 MPa at the licensing temperature limit. Results also show that the limiting case for hoop stress may not be at the highest rod internal pressure in all cases but will be related to the axial temperature and oxidation profiles of the rods at the end of life and in storage.« less
McAbee, Kathryn; Reinhardt, Keith; Germino, Matthew; Bosworth, Andrew
2017-01-01
Semi-arid rangelands are important carbon (C) pools at global scales. However, the degree of net C storage or release in water-limited systems is a function of precipitation amount and timing, as well as plant community composition. In northern latitudes of western North America, C storage in cold-desert ecosystems could increase with boosts in wintertime precipitation, in which climate models predict, due to increases in wintertime soil water storage that enhance summertime productivity. However, there are few long-term, manipulative field-based studies investigating how rangelands will respond to altered precipitation amount or timing. We measured aboveground C pools and fluxes at leaf, soil, and ecosystem scales over a single growing season in plots that had 200 mm of supplemental precipitation added in either winter or summer for the past 21 years, in shrub- and exotic-bunchgrass-dominated garden plots. At our cold-desert site (298 mm precipitation during the study year), we hypothesized that increased winter precipitation would stimulate the aboveground C uptake and storage relative to ambient conditions, especially in plots containing shrubs. Our hypotheses were generally supported: ecosystem C uptake and long-term biomass accumulation were greater in winter- and summer-irrigated plots compared to control plots in both vegetation communities. However, substantial increases in the aboveground biomass occurred only in winter-irrigated plots that contained shrubs. Our findings suggest that increases in winter precipitation will enhance C storage of this widespread ecosystem, and moreso in shrub- compared to grass-dominated communities.
McAbee, Kathryn; Reinhardt, Keith; Germino, Matthew J; Bosworth, Andrew
2017-03-01
Semi-arid rangelands are important carbon (C) pools at global scales. However, the degree of net C storage or release in water-limited systems is a function of precipitation amount and timing, as well as plant community composition. In northern latitudes of western North America, C storage in cold-desert ecosystems could increase with boosts in wintertime precipitation, in which climate models predict, due to increases in wintertime soil water storage that enhance summertime productivity. However, there are few long-term, manipulative field-based studies investigating how rangelands will respond to altered precipitation amount or timing. We measured aboveground C pools and fluxes at leaf, soil, and ecosystem scales over a single growing season in plots that had 200 mm of supplemental precipitation added in either winter or summer for the past 21 years, in shrub- and exotic-bunchgrass-dominated garden plots. At our cold-desert site (298 mm precipitation during the study year), we hypothesized that increased winter precipitation would stimulate the aboveground C uptake and storage relative to ambient conditions, especially in plots containing shrubs. Our hypotheses were generally supported: ecosystem C uptake and long-term biomass accumulation were greater in winter- and summer-irrigated plots compared to control plots in both vegetation communities. However, substantial increases in the aboveground biomass occurred only in winter-irrigated plots that contained shrubs. Our findings suggest that increases in winter precipitation will enhance C storage of this widespread ecosystem, and moreso in shrub- compared to grass-dominated communities.
NASA Astrophysics Data System (ADS)
Matzanke, B. F.; Bill, E.; Trautwein, A. X.; Winkelmann, G.
1990-07-01
In the yeasts Rhodotorula minuta and Ustilago sphaerogena siderophores represent the main intracellular iron pool. We suggest a ferritin substituting function of these siderophores in addition to their role as iron transport agents. In Rhodotorula transport and storage siderophore is the same compound whereas in Ustilago the iron-storage siderophore is ferrichrome. Besides siderophores, merely two iron metabolites can be observed. Other iron-requiring compounds are at least one order of magnitude less abundant in these yeasts. The ferrous metabolite has been detected in many other microbial systems and seems to be of general occurence and importance.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Stepanov, Vyacheslav; Potapov, Victor; Safronov, Alexey
2013-07-01
The underwater spectrometric system for survey the bottom of material science multi-loop reactor MR ponds was elaborated. This system uses CdZnTe (CZT) detectors that allow for spectrometric measurements in high radiation fields. The underwater system was used in the spectrometric survey of the bottom of the MR reactor pool, as well as in the survey located in the MR storage pool of highly radioactive containers and parts of the reactor construction. As a result of these works irradiated nuclear fuel was detected on the bottom of pools, and obtained estimates of the effective surface activity detected radionuclides and created bymore » them the dose rate. (authors)« less
A&M. TAN607. Section views of hot shop. Section E shows ...
A&M. TAN-607. Section views of hot shop. Section E shows equipment areas along rear wall. Section F shows storage pool cut along east/west line. Roof trusses, shelves along sides of pool, drain, roof trusses, shelves along sides of pool, drain, and sump. Section G cuts along north/south to show centerline of turntables, manipulator arms, O-man bridge, crane bridge. Referent drawing is ID-33-E-158 above. Ralph M. Parsons 902-3-ANP-607-A 107. Date: December 1952, but as-built in 1982. Approved by INEEL Classification Office for public release. INEEL index code no. 034-0607-00-693-106759 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID
Dessels, Carla; Durandt, Chrisna; Pepper, Michael S
2018-03-19
Pooled human platelet lysate (pHPL) has been used to expand adipose-derived stromal cells (ASCs) and can be formulated using fresh or expired buffy coats (BCs) which are then resuspended in either plasma or an additive solution. Not much is known about the effects that expired products and additive solutions have on ASC expansion, and the need for quality control and release criteria has been expressed. This pilot study compared proliferation, cell size, morphology and immunophenotype of ASCs expanded in the different pHPL alternatives versus foetal bovine serum (FBS). Quality control criteria were assessed prior to and during the manufacture of the pHPL alternatives. ASCs were then expanded in 1%, 2.5%, 5% or 10% of the different pHPL alternatives or in 10% FBS. Cell size, morphology, cell number and immunophenotype were measured using microscopy and flow cytometry. The majority of the pHPL alternatives were within the recommended ranges for the quality control criteria. ASCs expanded in the pHPL alternatives were smaller in size, displayed a tighter spindle-shaped morphology, increased cell growth and had a similar immunophenotype (with the exception of CD34 and CD36) when compared to ASCs expanded in FBS. Here we report on the effects that expired BC products and additive solutions have on ASC expansion. When taken together, our findings indicate that all of the pHPL alternatives can be considered to be suitable replacements for FBS for ASC expansion, and that expired BC products can be used as an alternative to fresh BC products.
NASA Astrophysics Data System (ADS)
Sala, A.; Martínez-Vilalta, J.; Asencio, M.; Lloret, F.; Palacio, S.; Galiano, L.; Hoch, G.; Piper, F.
2013-12-01
Forests store significant amounts of C globally and recent reports of forest mortality world-wide have generated strong concern. Evidence suggests that increasing drought associated with climate change is a primary cause of tree stress and subsequent mortality. This has generated an urgent need to predict how forests will cope with increasing stress. Storage of non-structural C compounds (NSC, compounds not permanently invested in structural biomass that can later be used to support diverse plant functions) is critical for survival during periods when C assimilation does not meet demand. However, remarkable knowledge gaps exist to accurately predict plant growth and survival under climate change. Although trees accumulate relatively large pools of NSC, there is a strong debate on how these pools build up over time. On the one hand, it is frequently assumed that the build- up of NSC in trees occurs when supply via photosynthesis exceeds overall demands. If so, the abundant NSC pools in trees reflect an overabundance of C in the long term. An alternative explanation is that trees regulate NSC storage to maintain sufficient pools to cope with asynchronies between demand and supply and with stresses that long lived plants inevitably experience during their life time. However, our understanding of whether and how trees regulate storage in the long term is minimal. Here, we assembled a new global database to examine broad patterns of seasonal NSC variation across organs, life forms and biomes, and the degree to which NSC storage is depleted in plants under a wide range of natural conditions. We compiled seasonal data (at least three measurements over a minimum of four months) for ca. 200 wild species under natural conditions. On average, NSC account for ca. 8-10% of dry plant biomass. NSC and starch concentrations do not vary significantly with biome, but soluble sugars (SS) in plants from Mediterranean biomes are higher than in temperate or tropical biomes. On average, seasonal NSC minimums are between 50 and 60% of the average, indicating that plants rarely deplete NSC. NSC, starch and SS vary strongly seasonally, with a strong depletion of SS during the growing season and a general increase during winter months, particularly in boreal and temperate biomes. In Mediterranean biomes, there is a second SS peak during the dry period. Importantly, with the exception of tropical biomes, the decrease of SS during the growing season is generally accompanied with increases in starch (especially in stems and leaves) despite constant or even decreasing NSC concentrations. These results suggest that during the growing season plants maintain or accumulate reserve compounds, even under high growth demand. Our results tentatively suggest that, on a seasonal basis, plants put aside storage C compounds during periods of maximum growth. Our results are also consistent with an important role of carbon storage in osmotic regulation to cope with winter cold and summer drought.
Modeling the effects of isolated wetlands on downstream hydrology at the watershed scale
Geographically-isolated wetlands, wetlands completely surrounded by uplands (e.g., prairie potholes, vernal pools and cypress domes), are depressional landscape features. They provide numerous ecological functions including biogeochemical cycling and water storage and recharge an...
A&M. TAN607. Construction view, facing southwest. At upper left of ...
A&M. TAN-607. Construction view, facing southwest. At upper left of view, north-wall equipment and operating galleries take shape on hot shop. Pumice-block side of storage pool section in center left of view. Water filter building (TAN-608) next to north wall of pool. Hot liquid waste building (TAN-616) at right of view. Note concrete construction of TAN-608 and 616. Date: January 18, 1954. INEEL negative no. 9604 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID
Konarska, Agata
2013-06-01
The structure of fruit peel of two apple varieties 'Szampion' and 'Jonagold' was investigated using light microscopy as well as scanning and transmission electron microscopy. The samples were taken immediately after harvest and after 6-month controlled atmosphere storage. The Szampion and Jonagold fruit differed in terms of the surface type, number of lenticels, thickness of the cuticular epithelium, height of epidermal cells and thickness of the hypodermis as well as the amount of crystalline wax and the number of microcracks formed on the fruit surface. The 6-month storage resulted in fruit weight loss, increased numbers and depth of microcracks, thickening of the amorphous wax layer and enhanced production of platelet forms of crystalline wax, which filled the microcracks abundantly. Compared with Jonagold, the Szampion fruit exhibited a fewer lenticels, a bigger number of microcracks, smaller amounts of crystalline wax and more substantial weight loss. The apple varieties studied had a reticulate-lamellate cuticle, and at harvest, the epidermal and hypodermal cells contained numerous amyloplasts filled with starch grains, which were not found after the storage period. Additionally, after storage, the cell protoplasts in the apple peel displayed a disorganised structure, and their vacuoles contained fragments of cell membranes, intravacuolar precipitates and deposits, and spherical bodies. The results may facilitate better understanding of changes occurring in fruits of Szampion and Jonagold during storage and help choose the best storage conditions to reduce loss of weight and prevent impairment of fruit quality.
NASA Astrophysics Data System (ADS)
Dou, Xiaolin; He, Ping; Cheng, Xiaoli; Zhou, Wei
2016-01-01
Quantification of dynamics of soil organic carbon (SOC) pools under the influence of long-term fertilization is essential for predicting carbon (C) sequestration. We combined soil chemical fractionation with stable C isotope analyses to investigate the C dynamics of the various SOC pools after 25 years of fertilization. Five types of soil samples (0-20, 20-40 cm) including the initial level (CK) and four fertilization treatments (inorganic nitrogen fertilizer, IN; balanced inorganic fertilizer, NPK; inorganic fertilizer plus farmyard manure, MNPK; inorganic fertilizer plus corn straw residue, SNPK) were separated into recalcitrant and labile fractions, and the fractions were analysed for C content, C:N ratios, δ13C values, soil C and N recalcitrance indexes (RIC and RIN). Chemical fractionation showed long-term MNPK fertilization strongly increased the SOC storage in both soil layers (0-20 cm = 1492.4 gC m2 and 20-40 cm = 1770.6 gC m2) because of enhanced recalcitrant C (RC) and labile C (LC). The 25 years of inorganic fertilizer treatment did not increase the SOC storage mainly because of the offsetting effects of enhanced RC and decreased LC, whereas no clear SOC increases under the SNPK fertilization resulted from the fast decay rates of soil C.
Dou, Xiaolin; He, Ping; Cheng, Xiaoli; Zhou, Wei
2016-01-11
Quantification of dynamics of soil organic carbon (SOC) pools under the influence of long-term fertilization is essential for predicting carbon (C) sequestration. We combined soil chemical fractionation with stable C isotope analyses to investigate the C dynamics of the various SOC pools after 25 years of fertilization. Five types of soil samples (0-20, 20-40 cm) including the initial level (CK) and four fertilization treatments (inorganic nitrogen fertilizer, IN; balanced inorganic fertilizer, NPK; inorganic fertilizer plus farmyard manure, MNPK; inorganic fertilizer plus corn straw residue, SNPK) were separated into recalcitrant and labile fractions, and the fractions were analysed for C content, C:N ratios, δ(13)C values, soil C and N recalcitrance indexes (RIC and RIN). Chemical fractionation showed long-term MNPK fertilization strongly increased the SOC storage in both soil layers (0-20 cm = 1492.4 gC m(2) and 20-40 cm = 1770.6 gC m(2)) because of enhanced recalcitrant C (RC) and labile C (LC). The 25 years of inorganic fertilizer treatment did not increase the SOC storage mainly because of the offsetting effects of enhanced RC and decreased LC, whereas no clear SOC increases under the SNPK fertilization resulted from the fast decay rates of soil C.
[Fever-shivers reaction and standard platelet concentrates transfusion: a prospective study].
Houissa, B; Abdelkefi, S; Bouslama, M; Zaeir, M; Chakroun, T; Ghachem, L; Yacoub, S
2003-09-01
Fever-shivers reaction (FSR) is the most frequent transfusion immediate incident related to platelet transfusions. The aim of our prospective study was to assess the frequency of the different immediate incidents, especially the frequency and the causes of the FSR, observed during the transfusion of standard platelet concentrates (SPC). For each FSR, analysis of causes included: a bacterial culture of the implicated SPC, a blood culture and HLA antibody screening (lymphocytotoxicity assay) among the patients. In the study period, 34 patients were followed during 74 transfusions. Ten immediate incidents were noted; FSR: N = 8, erythema-urticaria: N = 1 and nausea-vomit: N = 1. The FSR was observed in 6 patients who received 56 SPC. Analysis of causes of this reaction revealed that: HLA antibodies were present in one patient; bacterial contamination was not found neither among the patients nor in the implicated SPC, and the risk of the FSR occurrence rose with increased storage time of the SPC transfused. Indeed, a significant difference was noted between the mean age of the SPC implicated in the FSR and the mean age of those not implicated (P = 0,0028). In conclusion, the FSR is a frequent incident observed during SPC transfusions. In the majority of cases, the cause of this reaction was not identified. Further studies will be necessary to better understand the physiological mechanisms of the FSR.
Hitzler, Walter E
2014-01-01
The transfusion efficacy of ATK, which contain fully functional platelets, is beyond all doubt. The equivalence of ATK and PTK has been subject of many studies. Some of those studies show the superiority of ATK's, while others do not, but there have been no studies that demonstrated a superiority of PTK's. The superiority of platelets stored in plasma and in third generation additive solution was demonstrated in clinical studies; therefore, it cannot be said that all the platelet concentrates on the German market are equivalent in efficacy. Of decisive importance, above all, is the risk of transfusion-transmitted infections with known pathogens, or those not yet discovered. This risk is different for ATK compared to PTK. Taking this difference in risk and the difference in donor exposure of transfused patients into account, it can definitely be said that ATK and PTK are not equivalent. In 2012, the Robert-Koch-Institute (RKI) published a mathematical risk model for different platelet concentrates and assessed the risk of transmitting known pathogens such as HIV, HCV, and HBV. The risk was higher for PTK compared to ATK. The relative risks for PTK derived from 4BCs were 2.2 (95%--CI: 2.1-2.4) for HIV, 2.7 (95%--CI: 2.5-3.0) for HCV, and 2.2 (95%--CI: 2.8-3.7) for HBV. At the present time, these are the relative risks of transfusion-transmitted infections with the traditional pathogens for PTK compared to ATK. In addition to the RKI assessed risks, there is the theoretical risk of a new, unknown agent, transmitted through blood exposure. The magnitude of this risk is hardly predictable for PTK. The experience gathered so far, especially in the last three decades, with the emergence of HIV, prions, and West Nil virus, shows that the biological nature of a next transfusion-transmissible infectious agent cannot be predictable. This agent, if we think at a conventional sexually transmissible agent with nucleic acid and long latent period, would spread first in areas with high population density and thereby reduce the theoretical advantage of ATK (but definitely would not nullify it!). It is equally plausible, however, that this agent would behave like a prion, non-sexual transmission, or like a West-Nil virus, a non-contagious vector-transmitted agent. For PTK this would mean a relative risk up to 4 times (PTK from 4 BCs) or 5 times (PTK from 5 BCs) higher than the risk estimated by the Robert-Koch-Institute. If, taking the passive surveillance data and the changing variables (donor frequency, donor population, and donor location) into account, the risk of transmission of an infection via ATK (exposure to 1 donor) with HIV, HCV, and HBV moves closer to the higher risk of PTK (exposure to 4 or 8 donors, in case of double ATK per patient), this result of the risk model calculation by no means indicates any equivalency between PTK and ATK with respect to the risk of transmission of infection. The modifiable variables of donor frequency, donor population, and donor location need to be modified, as scientific deductions, in such a way that the avoidable risk of ATK which is influenced by these variables can be corrected to the minimum risk of a transmission of infection of HIV, HBV, and HCV via ATK in comparison to PTK. The minimum risk of a possible transmission of infection via ATK (exposure to 1 donor) is the basic intrinsic risk of each individual blood donation. The basic intrinsic risk increases relative to the number of blood donations or exposure to donors (PtK has an unalterable, production-dependent exposure to 4 or 8 donors). Let us consider a 1:1.000 prevalence for a new pathogen, which is spread equally in each donor population (apheresis and whole blood) and the present case of approximately 500,000 transfused platelet concentrates in Germany. This means that for the production of 4 PTK about 2 million donations are processed, 2,000 infectious Buffy-Coats are obtained and, thereby, 2,000 infectious PTK. In the case of ATK, considering five (5) donations per year, theoretically, it would mean 100 donors infected and 500 infectious ATK. Considering 15 apheresis donations per donor per year, this would mean that 33 donors are infected, but still 500 infectious ATK would be produced. The prion is an example of a pathogen that, although its existence is well known, cannot be proven or pathogen-reduced. In addition, it has a very long incubation period compared to the donation intervals. Due to the manufacturing process, PTK has a 4-fold higher donor exposure and therefore a 4-fold higher risk for transfusion-transmitted infections compared to ATK. If a patient needs the transfusion of two platelet concentrates, by transfusing a double-ATK from the same donor the risk of transfusion-transmitted infections will remain the same. On the other hand, the risk will increase by 8-fold by transfusing two PTK. The only current possibility to prevent or to minimize the risk of infection with prions is to minimize the donor exposure by transfusing ATK instead of PTK. Hypothetical risk scenarios carry significant weight in law. This can be seen in the constant rulings of the German Federal Supreme Court (Bundesgerichtshofs (BGH)) on the so-called hypothetical risk explanations (BGH, NJW 1996, 776, 777; 2000, 1784, 1787; 2005, 2614, 2616). Therefore, a risk does not need to be confirmed to be subject to compulsory explanation. It is sufficient that serious voices in the medical scientific community point to specific risks, which cannot be set aside as insignificant outside opinions, but must be viewed as serious warnings. According to the rulings, patients must even be informed of rare and often extremely rare risks, which could, should they come true, significantly impact daily life and, despite their rarity, are specific to the treatment and are startling for the ordinary person (BGH, 15.02.2000- VI ZR 48199 -; BGH, 30.11.2004 - VI ZR 209104 -; OLG Hamm, 29.09.2010 - 1-3 V 169109). These conditions have been fulfilled for PTK according to current knowledge, especially since, in the meantime in several rulings, the federal supreme court has required the reference to as yet unknown risks (refer to BGH, 13.06.2006 - VI ZR 323104 - for the use of new medical treatment methods, BGH, 27.06.2007 - VI ZR 55105 for experimental therapy using new, unapproved medication BGH, 06.07.2010 - VI ZR 198109 - for unknown risks cannot be excluded, for example based on anatomical conditions). ATK and PTK are therapeutic alternatives with the same range of indications for treatment using thrombocytes, however, with differing risks of infection, with different exposures to donors, and with different efficacy. ATK and PTK. ATK and PTK are therapeutic alternatives in terms of pharmaceutical law based on the different risks and the different quality. Patients must be informed of therapeutic alternatives such as ATK and PTK according to the patient rights law. Denial of reimbursement for additional fees for ATK by individual insurance companies (or paying authorities) deviates blatantly, as seen in the ruling of the Social Court of of the Saarland in this matter, from the basic requirement of the Transfusion Law (Transfusionsgesetz (TFG)) and is legally incorrect. The legality of the question whether the transfusion of ATK is indicated or if PTK had sufficed, is not allowable within the context of an MDK-Test according to subsection 275 ff. SGB V. The denial is a direct infringment on the treatment authority of the attending hospital physician and is illegal according to subsection 275 Abs. 5 SGBV. It is certainly possible to establish a full ATK supply and can be immediately realized by increasing donation rates from 5 to 8.3 apheresis donations per year in the current scenario of apheresis structure and donor population. The donation interval between two apheresis donations would be 49 days. A complete supply with ATK can also be immediately implemented by enlarging the donor population, keeping the current apheresis donation frequency. The donor pool must be increased by 24,576 donors, which means a 67% increase of the existing donor population. A transition to an ATK supply that can cover the entire demand can certainly be realized in a short period of time, while assuring a complete supply with PTK is not a realistic option. All existing studies advise taking extreme caution with any alternative to the current German gold standard for the treatment of hyporegenerative thrombocytopenia. A prophylactic transfusion of a non-pathogen-inactivated platelet concentrate with on average 3 x 10(11) platelets is recommended when the platelet count drops below the threshold of 10,000/microL. All other alternatives to this strategy show an increase in intracranial bleeding events. The existing studies on platelet dose (PLADO-Trial and StoP-Trial) do not recommend deviating from 3 x 10(11) platelets per unit. On the contrary, these studies demonstrate that the only practicable way is to individually correlate every platelet transfusion to the patient body surface. Considering the current knowledge, it is not justified to lower the standard dose and, for certain patient groups, to switch from prophylaxis to therapeutic platelet transfusion. Applying ATK or PTK with a lower platelet content and only for therapeutic purposes, could considerably increase the bleeding risk, especially for WHO grades III and IV. This will also affect all the patients who receive an induction treatment. Through pathogen reduction, in parallel with platelet loss (Apoptosis), the function of the treated platelets is impaired. Alternatively, the cell destruction caused during this process could result in a release of platelet microRNA directly into the supernatant or in microvesicles. This reduction of microRNA will affect the storage of the platelets. (ABSTRACT TRUNCATED)
Impact of Oral Fluid Collection Device on Cannabinoid Stability Following Smoked Cannabis
Anizan, Sébastien; Bergamaschi, Mateus M.; Barnes, Allan J.; Milman, Garry; Desrosiers, Nathalie; Lee, Dayong; Gorelick, David A.; Huestis, Marilyn A.
2014-01-01
Evaluation of cannabinoid stability in authentic oral fluid (OF) is critical, as most OF stability studies employed fortified or synthetic OF. Participants (n=16) smoked a 6.8% delta-9-tetrahydrocannabinol (THC) cigarette, and baseline concentrations of THC, 11-nor-9-carboxy-THC (THCCOOH), cannabidiol (CBD), and cannabinol (CBN) were determined within 24h in 16 separate pooled samples (collected 1h before to 10.5 or 13h after smoking). OF was collected with the StatSure Saliva Sampler™ and Oral-Eze® devices. Oral-Eze samples were re-analyzed after room temperature (RT) storage for 1 week, and for both devices after 4°C for 1 and 4 weeks, and –20°C for 4 and 24 weeks. Concentrations ±20% from initial concentrations were considered stable. With the StatSure device, all cannabinoids were within 80-120% median %baseline for all storage conditions. Individual THC, CBD, CBN and THCCOOH pool concentrations were stable in 100%, 100%, 80-94% and >85%, respectively, across storage conditions. With the Oral-Eze device, at RT or refrigerated storage (for 1 and 4 weeks), THC, CBD and THCCOOH were stable in 94-100%, 78-89% and 93-100% of samples, respectively, while CBN concentrations were 53–79% stable. However, after 24 weeks at -20°C, stability decreased, especially for CBD, with a median of 56% stability. Overall, the collection devices’ elution/stabilizing buffers provided good stability for OF cannabinoids, with the exception of the more labile CBN. To ensure OF cannabinoid concentration accuracy, these data suggest analysis within 4 weeks at 4°C storage for Oral-Eze collection and within 4 weeks at 4°C or 24 weeks at -20°C for StatSure collection. PMID:24995604
Fires in storages of LFO: Analysis of hazard of structural collapse of steel-aluminium containers.
Rebec, A; Kolšek, J; Plešec, P
2016-04-05
Pool fires of light fuel oil (LFO) in above-ground storages with steel-aluminium containers are discussed. A model is developed for assessments of risks of between-tank fire spread. Radiative effects of the flame body are accounted for by a solid flame radiation model. Thermal profiles evolved due to fire in the adjacent tanks and their consequential structural response is pursued in an exact (materially and geometrically non-linear) manner. The model's derivation is demonstrated on the LFO tank storage located near the Port of Koper (Slovenia). In support of the model, data from literature are adopted where appropriate. Analytical expressions are derived correspondingly for calculations of emissive characteristics of LFO pool fires. Additional data are collected from experiments. Fire experiments conducted on 300cm diameter LFO pans and at different wind speeds and high-temperature uniaxial tension tests of the analysed aluminium alloys types 3xxx and 6xxx are presented. The model is of an immediate fire engineering practical value (risk analyses) or can be used for further research purposes (e.g. sensitivity and parametric studies). The latter use is demonstrated in the final part of the paper discussing possible effects of high-temperature creep of 3xxx aluminium. Copyright © 2015 Elsevier B.V. All rights reserved.
Stimulation of terrestrial ecosystem carbon storage by nitrogen addition: a meta-analysis
Yue, Kai; Peng, Yan; Peng, Changhui; Yang, Wanqin; Peng, Xin; Wu, Fuzhong
2016-01-01
Elevated nitrogen (N) deposition alters the terrestrial carbon (C) cycle, which is likely to feed back to further climate change. However, how the overall terrestrial ecosystem C pools and fluxes respond to N addition remains unclear. By synthesizing data from multiple terrestrial ecosystems, we quantified the response of C pools and fluxes to experimental N addition using a comprehensive meta-analysis method. Our results showed that N addition significantly stimulated soil total C storage by 5.82% ([2.47%, 9.27%], 95% CI, the same below) and increased the C contents of the above- and below-ground parts of plants by 25.65% [11.07%, 42.12%] and 15.93% [6.80%, 25.85%], respectively. Furthermore, N addition significantly increased aboveground net primary production by 52.38% [40.58%, 65.19%] and litterfall by 14.67% [9.24%, 20.38%] at a global scale. However, the C influx from the plant litter to the soil through litter decomposition and the efflux from the soil due to microbial respiration and soil respiration showed insignificant responses to N addition. Overall, our meta-analysis suggested that N addition will increase soil C storage and plant C in both above- and below-ground parts, indicating that terrestrial ecosystems might act to strengthen as a C sink under increasing N deposition. PMID:26813078
High energy neutron transmission analysis of dry cask storage
NASA Astrophysics Data System (ADS)
Greulich, Christopher; Hughes, Christopher; Gao, Yuan; Enqvist, Andreas; Baciak, James
2017-12-01
Since the U.S. currently only approves of storing used nuclear fuel in pools or dry casks, the demand for dry cask storage is on the rise due to the continuous operation of currently existing nuclear plants which are reaching or have reached the capacity of their used fuel pools. With the rising demand comes additional pressure to ensure the integrity of dry cask systems. Visual inspection is costly and man-power intensive, so alternative nondestructive testing techniques are desired to insure the continued safe and effective storage of fuel. One such approach being investigated by the University of Florida is neutron based computed tomography. Simulations in MCNP are preformed where D-T energy neutrons are transmitted through the dry cask and measured on the opposite side. If the transmitted signal is clear enough, the interior of the cask can be reconstructed from the measurement of the alterations of neutron signal intensity using standard mathematical techniques developed for medical imaging. Preliminary efforts show a correlation between energy and number of scatters (which is an indication of retention of position information). Work is ongoing to quantify if the correlation is strong enough that an energy discriminator may be used as a filter in future image reconstruction. The calculated transmission probability suggests that an image could be reconstructed with a week of scanning.
Shading responses of carbon allocation dynamics in mountain grassland
NASA Astrophysics Data System (ADS)
Bahn, M.; Lattanzi, F. A.; Brueggemann, N.; Siegwolf, R. T.; Richter, A.
2012-12-01
Carbon (C) allocation strongly influences plant and soil processes. Global environmental changes can alter source - sink relations of plants with potential implications for C allocation. Short-term C allocation dynamics in ecosystems and their responses to environmental changes are still poorly understood. To analyze effects of assimilate supply (i.e. C source strength) on ecosystem C allocation dynamics and the role of non-structural carbohydrates, canopy sections of a mountain meadow were pulse labeled with 13CO2 and subsequently shaded for a week or left unshaded (control). Tracer dynamics in above- and belowground sucrose and starch pools were analysed and coupled using compartmental modelling. The hypothesis was tested that shading affects tracer dynamics in non-structural carbohydrates and diminishes the transfer of recently assimilated C to roots and their storage pools. In unshaded plots up to 40% of assimilated C was routed through short-term storage in shoot starch and sucrose to buffer day / night cycles in photosynthesis. Shoot- and root sucrose and shoot starch were kinetically closely related pools. The tracer dynamics of the modelled root sucrose pool corresponded well with those in soil CO2 efflux. Root starch played no role in buffering day / night cycles and likely acted as a seasonal store. Shading strongly reduced sucrose and starch concentrations in shoots but not roots and resulted in a massive reduction of leaf respiration, while root respiration was much less diminished. Shading affected tracer dynamics in sucrose and starch of shoots: shoot starch rapidly lost tracer, while sucrose transiently increased its tracer content. Surprisingly, shading did not alter the dynamics of root carbohydrates. Even under severe C limitation after one week of shading, tracer C continued to be incorporated in root starch. Also the amount of 13C incorporated in phospholipid fatty acids of soil microbial communities was not reduced by shading, though its residence time followed a changed pattern, suggesting an influence of C source strength on the utilization and turnover of recent plant-derived C. These findings will be discussed in the broader context of plant and ecosystem carbon allocation, with particular reference to the concepts of 'source versus sink strength' and 'passive versus active C storage'.
Optimising LAN access to grid enabled storage elements
NASA Astrophysics Data System (ADS)
Stewart, G. A.; Cowan, G. A.; Dunne, B.; Elwell, A.; Millar, A. P.
2008-07-01
When operational, the Large Hadron Collider experiments at CERN will collect tens of petabytes of physics data per year. The worldwide LHC computing grid (WLCG) will distribute this data to over two hundred Tier-1 and Tier-2 computing centres, enabling particle physicists around the globe to access the data for analysis. Although different middleware solutions exist for effective management of storage systems at collaborating institutes, the patterns of access envisaged for Tier-2s fall into two distinct categories. The first involves bulk transfer of data between different Grid storage elements using protocols such as GridFTP. This data movement will principally involve writing ESD and AOD files into Tier-2 storage. Secondly, once datasets are stored at a Tier-2, physics analysis jobs will read the data from the local SE. Such jobs require a POSIX-like interface to the storage so that individual physics events can be extracted. In this paper we consider the performance of POSIX-like access to files held in Disk Pool Manager (DPM) storage elements, a popular lightweight SRM storage manager from EGEE.
Neutrophil Kinetics in Acute Infection*
Marsh, J. C.; Boggs, D. R.; Cartwright, G. E.; Wintrobe, M. M.
1967-01-01
Neutrophil kinetics of acute experimental infection were studied with diisopropylfluorophosphate-32P labeling in 31 dogs inoculated intrabronchially with pneumococci. In vitro neutrophil labeling indicated a rapid transit time through the blood in early infections, with an elevated marginal granulocyte pool sometimes preceding an elevation of the circulating granulocyte pool. 13 hr after infection, the circulating and total blood granulocyte pools were increased but the rate of neutrophil transit through the blood was normal. During the recovery from infection there was a marked prolongation of neutrophil blood transit time, suggesting virtually complete cessation of bone marrow release of neutrophils into the blood. Labeling of neutrophils in vivo indicated an increased rate of emptying of the bone marrow storage pool proportional to the severity of infection as measured by the fever index. The change in the blood ratio of nonsegmented to segmented neutrophils was a much more accurate index of the severity of infection than the blood granulocyte concentration, correlating significantly with the fever index. PMID:6073999
Zainathan, S C; Carson, J; Crane, M St J; Nowak, B F
2013-04-01
The use of swabs relative to organs as a sample collection method for the detection of Tasmanian salmon reovirus (TSRV) in farmed Tasmanian Atlantic salmon, Salmo salar L., was evaluated by RT-qPCR. Evaluation of individual and pooled sample collection (organs vs swabs) was carried out to determine the sensitivity of the collection methods and the effect of pooling of samples for the detection of TSRV. Detection of TSRV in individual samples was as sensitive when organs were sampled compared to swabs, and in pooled samples, organs demonstrated a sensitivity of one 10-fold dilution higher than sampling of pooled swabs. Storage of swabs at 4 °C for t = 24 h demonstrated results similar to those at t = 0. Advantages of using swabs as a preferred sample collection method for the detection of TSRV compared to organ samples are evident from these experimental trials. © 2012 Blackwell Publishing Ltd.
The Camp Caretaker: A Hidden Treasure.
ERIC Educational Resources Information Center
Ezersky, Eugene M.
1996-01-01
At a round-table discussion, five camp caretakers identified common camp maintenance problems. Snow loads, wooden floors, storage of lake equipment, removal of grass cuttings and leaves, local suppliers, vandalism and trespassing, swimming pools, assigning work, use of outside contractors, decisions to replace or repair, job satisfaction, and…
Weymann, Alexander; Ali-Hasan-Al-Saegh, Sadeq; Sabashnikov, Anton; Popov, Aron-Frederik; Mirhosseini, Seyed Jalil; Liu, Tong; Lotfaliani, Mohammadreza; de Oliveira Sá, Michel Pompeu Barros; Baker, William L.; Yavuz, Senol; Zeriouh, Mohamed; Jang, Jae-Sik; Dehghan, Hamidreza; Meng, Lei; Testa, Luca; D’Ascenzo, Fabrizio; Benedetto, Umberto; Tse, Gary; Nombela-Franco, Luis; Dohmen, Pascal M.; Deshmukh, Abhishek J.; Linde, Cecilia; Biondi-Zoccai, Giuseppe; Stone, Gregg W.; Calkins, Hugh
2017-01-01
Background Atrial fibrillation (AF) is one of the most critical and frequent arrhythmias precipitating morbidities and mortalities. The complete blood count (CBC) test is an important blood test in clinical practice and is routinely used in the workup of cardiovascular diseases. This systematic review with meta-analysis aimed to determine the strength of evidence for evaluating the association of hematological parameters in the CBC test with new-onset and recurrent AF. Material/Methods We conducted a meta-analysis of observational studies evaluating hematologic parameters in patients with new-onset AF and recurrent AF. A comprehensive subgroup analysis was performed to explore potential sources of heterogeneity. Results The literature search of all major databases retrieved 2150 studies. After screening, 70 studies were analyzed in the meta-analysis on new-onset AF and 23 studies on recurrent AF. Pooled analysis on new-onset AF showed platelet count (PC) (weighted mean difference (WMD)=WMD of −26.39×109/L and p<0.001), mean platelet volume (MPV) (WMD=0.42 FL and p<0.001), white blood cell (WBC) (WMD=−0.005×109/L and p=0.83), neutrophil to lymphocyte ratio (NLR) (WMD=0.89 and p<0.001), and red blood cell distribution width (RDW) (WMD=0.61% and p<0.001) as associated factors. Pooled analysis on recurrent AF revealed PC (WMD=−2.71×109/L and p=0.59), WBC (WMD=0.20×109/L (95% CI: 0.08 to 0.32; p=0.002), NLR (WMD=0.37 and p<0.001), and RDW (WMD=0.28% and p<0.001). Conclusions Hematological parameters have significant ability to predict occurrence and recurrence of AF. Therefore, emphasizing the potential predictive role of hematological parameters for new-onset and recurrent AF, we recommend adding the CBC test to the diagnostic modalities of AF in clinical practice. PMID:28496093
Weymann, Alexander; Ali-Hasan-Al-Saegh, Sadeq; Sabashnikov, Anton; Popov, Aron-Frederik; Mirhosseini, Seyed Jalil; Liu, Tong; Lotfaliani, Mohammadreza; Sá, Michel Pompeu Barros de Oliveira; Baker, William L L; Yavuz, Senol; Zeriouh, Mohamed; Jang, Jae-Sik; Dehghan, Hamidreza; Meng, Lei; Testa, Luca; D'Ascenzo, Fabrizio; Benedetto, Umberto; Tse, Gary; Nombela-Franco, Luis; Dohmen, Pascal M; Deshmukh, Abhishek J; Linde, Cecilia; Biondi-Zoccai, Giuseppe; Stone, Gregg W; Calkins, Hugh; Surgery And Cardiology-Group Imcsc-Group, Integrated Meta-Analysis Of Cardiac
2017-05-12
BACKGROUND Atrial fibrillation (AF) is one of the most critical and frequent arrhythmias precipitating morbidities and mortalities. The complete blood count (CBC) test is an important blood test in clinical practice and is routinely used in the workup of cardiovascular diseases. This systematic review with meta-analysis aimed to determine the strength of evidence for evaluating the association of hematological parameters in the CBC test with new-onset and recurrent AF. MATERIAL AND METHODS We conducted a meta-analysis of observational studies evaluating hematologic parameters in patients with new-onset AF and recurrent AF. A comprehensive subgroup analysis was performed to explore potential sources of heterogeneity. RESULTS The literature search of all major databases retrieved 2150 studies. After screening, 70 studies were analyzed in the meta-analysis on new-onset AF and 23 studies on recurrent AF. Pooled analysis on new-onset AF showed platelet count (PC) (weighted mean difference (WMD)=WMD of -26.39×10^9/L and p<0.001), mean platelet volume (MPV) (WMD=0.42 FL and p<0.001), white blood cell (WBC) (WMD=-0.005×10^9/L and p=0.83), neutrophil to lymphocyte ratio (NLR) (WMD=0.89 and p<0.001), and red blood cell distribution width (RDW) (WMD=0.61% and p<0.001) as associated factors. Pooled analysis on recurrent AF revealed PC (WMD=-2.71×109/L and p=0.59), WBC (WMD=0.20×10^9/L (95% CI: 0.08 to 0.32; p=0.002), NLR (WMD=0.37 and p<0.001), and RDW (WMD=0.28% and p<0.001). CONCLUSIONS Hematological parameters have significant ability to predict occurrence and recurrence of AF. Therefore, emphasizing the potential predictive role of hematological parameters for new-onset and recurrent AF, we recommend adding the CBC test to the diagnostic modalities of AF in clinical practice.
Trojahn Kølle, Stig-Frederik; Oliveri, Roberto S; Glovinski, Peter V; Kirchhoff, Maria; Mathiasen, Anders Bruun; Elberg, Jens Jørgen; Andersen, Peter Stemann; Drzewiecki, Krzysztof Tadeusz; Fischer-Nielsen, Anne
2013-09-01
Because of an increasing focus on the use of adipose-derived stem cells (ASCs) in clinical trials, the culture conditions for these cells are being optimized. We compared the proliferation rates and chromosomal stability of ASCs that had been cultured in Dulbecco's modified Eagle's Medium (DMEM) supplemented with either pooled human platelet lysate (pHPL) or clinical-grade fetal bovine serum (FBS) (DMEM(pHPL) versus DMEM(FBS)). ASCs from four healthy donors were cultured in either DMEM(pHPL) or DMEM(FBS), and the population doubling time (PDT) was calculated. ASCs from two of the donors were expanded in DMEM(pHPL) or DMEM(FBS) and cultured for the final week before harvesting with or without the addition of vascular endothelial growth factor. We assessed the chromosomal stability (through the use of array comparative genomic hybridization), the expression of ASC and endothelial surface markers and the differentiation and angiogenic potential of these cells. The ASCs that were cultured in pHPL exhibited a significantly shorter PDT of 29.6 h (95% confidence interval, 22.3-41.9 h) compared with those cultured in FBS, for which the PDT was 123.9 h (95% confidence interval, 95.6-176.2 h). Comparative genomic hybridization analyses revealed no chromosomal aberrations. Cell differentiation, capillary structure formation and cell-surface marker expression were generally unaffected by the type of medium supplement that was used or by the addition of vascular endothelial growth factor. We observed that the use of pHPL as a growth supplement for ASCs facilitated a significantly higher proliferation rate compared with FBS without compromising genomic stability or differentiation capacity. Copyright © 2013 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.
Fragnoud, Romain; Flamand, Marie; Reynier, Frederic; Buchy, Philippe; Duong, Vasna; Pachot, Alexandre; Paranhos-Baccala, Glaucia; Bedin, Frederic
2015-11-14
Dengue is the most widespread mosquito-borne viral disease of public health concern. In some patients, endothelial cell and platelet dysfunction lead to life-threatening hemorrhagic dengue fever or dengue shock syndrome. Prognostication of disease severity is urgently required to improve patient management. The pathogenesis of severe dengue has not been fully elucidated, and the role of host proteins associated with viral particles has received little exploration. The proteomes of virion-enriched fractions purified from plasma pools of patients with dengue fever or severe dengue were compared. Virions were purified by ultracentrifugation combined with a water-insoluble polyelectrolyte-based technique. Following in-gel hydrolysis, peptides were analyzed by nano-liquid chromatography coupled to ion trap mass spectrometry and identified using data libraries. Both dengue fever and severe dengue viral-enriched fractions contained identifiable viral envelope proteins and host cellular proteins. Canonical pathway analysis revealed the identified host proteins are mainly involved in the coagulation cascade, complement pathway or acute phase response signaling pathway. Some host proteins were over- or under-represented in plasma from patients with severe dengue compared to patients with dengue fever. ELISAs were used to validate differential expression of a selection of identified host proteins in individual plasma samples of patients with dengue fever compared to patients with severe dengue. Among 22 host proteins tested, two could differentiate between dengue fever and severe dengue in two independent cohorts (olfactomedin-4: area under the curve (AUC), 0.958; and platelet factor-4: AUC, 0.836). A novel technique of virion-enrichment from plasma has allowed to identify two host proteins that have prognostic value for classifying patients with acute dengue who are more likely to develop a severe dengue. The impact of these host proteins on pathogenicity and disease outcome are discussed.
Siegel, Georg; Fleck, Erika; Elser, Stefanie; Hermanutz-Klein, Ursula; Waidmann, Marc; Northoff, Hinnak; Seifried, Erhard; Schäfer, Richard
2018-05-01
Endothelial colony-forming progenitor cells (ECFCs) are promising candidates for cell therapies. However, ECFC translation to the clinic requires optimized isolation and manufacture technologies according to good manufacturing practice (GMP). ECFCs were manufactured from steady-state peripheral blood (PB) leukapheresis (11 donors), using GMP-compliant technologies including pooled human platelet (PLT) lysate, and compared to human umbilical cord endothelial cells, human aortic endothelial cells, and human cerebral microvascular endothelial cells. Specific variables assessed were growth kinetics, phenotype, trophic factors production, stimulation of tube formation, and Dil-AcLDL uptake. ECFCs could be isolated from PB leukapheresis units with mean processed volume of 5411 mL and mean white blood cell (WBC) concentration factor of 8.74. The mean frequency was 1.44 × 10 -8 ECFCs per WBC, corresponding to a mean of 177.8 ECFCs per apheresis unit. Expandable for up to 12 cumulative population doublings, calculated projection showed that approximately 730 × 10 3 ECFCs could be manufactured from 1 apheresis unit. ECFCs produced epidermal growth factor, hepatocyte growth factor, vascular endothelial growth factor (VEGF)-A, PLT-derived growth factor-B, interleukin-8, and monocyte chemoattractant protein-1, featured high potential for capillary-like tubes formation, and showed no telomerase activity. They were characterized by CD29, CD31, CD44, CD105, CD117, CD133, CD144, CD146, and VEGF-R2 expression, with the most common subpopulation CD34+CD117-CD133-. Compared to controls, ECFCs featured greater Dil-AcLDL uptake and higher expression of CD29, CD31, CD34, CD44, CD144, and VEGF-R2. Here we show that isolation of ECFCs with proangiogenic profile from steady-state PB leukapheresis is feasible, marking a first step toward ECFC product manufacture according to GMP. © 2018 AABB.
Rosato, Mp; Iaffaldano, N
2011-04-01
As the preservation of the fertilizing capacity of rabbit spermatozoa for several days after semen collection remains a major target for the artificial insemination programs of rabbit breeding, a study was conducted to compare the efficacy of 5 or 15°C as holding temperature in lengthening the preservability of rabbit semen quality during 192 h of storage both in a solid (Cunigel) and a liquid (Tris-Citric acid-Glucose; TCG) extender. Six pooled semen samples (two ejaculates/male; two-three males/pool) were taken and made four aliquots: two aliquots were tenfold diluted with the TCG extender, whereas the other two were tenfold diluted with the Cunigel extender. One aliquot per diluent was stored at 5°C and the second one at 15°C. Sperm motility (light microscope), viability (SyBr-PI staining), plasma membrane functional integrity (Hypo-osmotic swelling test) and acrosome integrity (PSA-FITC staining) were recorded at 0, 48, 120 and 192 h of storage. In liquid-stored spermatozoa, mass motility and viability were significantly higher (p ≤ 0.05) in samples stored at 5°C than at 15°C at all the storage times; at 5°C resulted also higher (p ≤ 0.05) the percentages of both forward motility at 48 h and sperm functional integrity at 120 and 192 h of storage, whereas chilling temperature did not affect acrosome integrity. With the Cunigel extender, all the semen qualitative parameters were significantly higher in sample stored at 5 than 15°C over storage time (p ≤ 0.05); only acrosome integrity at 192 h was not different according to the chilling temperatures. In conclusion, 5°C were better than 15°C for the long-term storage of rabbit semen both in the TCG and Cunigel extender. © 2010 Blackwell Verlag GmbH.
Blood group genotyping: from patient to high-throughput donor screening.
Veldhuisen, B; van der Schoot, C E; de Haas, M
2009-10-01
Blood group antigens, present on the cell membrane of red blood cells and platelets, can be defined either serologically or predicted based on the genotypes of genes encoding for blood group antigens. At present, the molecular basis of many antigens of the 30 blood group systems and 17 human platelet antigens is known. In many laboratories, blood group genotyping assays are routinely used for diagnostics in cases where patient red cells cannot be used for serological typing due to the presence of auto-antibodies or after recent transfusions. In addition, DNA genotyping is used to support (un)-expected serological findings. Fetal genotyping is routinely performed when there is a risk of alloimmune-mediated red cell or platelet destruction. In case of patient blood group antigen typing, it is important that a genotyping result is quickly available to support the selection of donor blood, and high-throughput of the genotyping method is not a prerequisite. In addition, genotyping of blood donors will be extremely useful to obtain donor blood with rare phenotypes, for example lacking a high-frequency antigen, and to obtain a fully typed donor database to be used for a better matching between recipient and donor to prevent adverse transfusion reactions. Serological typing of large cohorts of donors is a labour-intensive and expensive exercise and hampered by the lack of sufficient amounts of approved typing reagents for all blood group systems of interest. Currently, high-throughput genotyping based on DNA micro-arrays is a very feasible method to obtain a large pool of well-typed blood donors. Several systems for high-throughput blood group genotyping are developed and will be discussed in this review.
Comparative analyses of industrial-scale human platelet lysate preparations.
Pierce, Jan; Benedetti, Eric; Preslar, Amber; Jacobson, Pam; Jin, Ping; Stroncek, David F; Reems, Jo-Anna
2017-12-01
Efforts are underway to eliminate fetal bovine serum from mammalian cell cultures for clinical use. An emerging, viable replacement option for fetal bovine serum is human platelet lysate (PL) as either a plasma-based or serum-based product. Nine industrial-scale, serum-based PL manufacturing runs (i.e., lots) were performed, consisting of an average ± standard deviation volume of 24.6 ± 2.2 liters of pooled, platelet-rich plasma units that were obtained from apheresis donors. Manufactured lots were compared by evaluating various biochemical and functional test results. Comprehensive cytokine profiles of PL lots and product stability tests were performed. Global gene expression profiles of mesenchymal stromal cells (MSCs) cultured with plasma-based or serum-based PL were compared to MSCs cultured with fetal bovine serum. Electrolyte and protein levels were relatively consistent among all serum-based PL lots, with only slight variations in glucose and calcium levels. All nine lots were as good as or better than fetal bovine serum in expanding MSCs. Serum-based PL stored at -80°C remained stable over 2 years. Quantitative cytokine arrays showed similarities as well as dissimilarities in the proteins present in serum-based PL. Greater differences in MSC gene expression profiles were attributable to the starting cell source rather than with the use of either PL or fetal bovine serum as a culture supplement. Using a large-scale, standardized method, lot-to-lot variations were noted for industrial-scale preparations of serum-based PL products. However, all lots performed as well as or better than fetal bovine serum in supporting MSC growth. Together, these data indicate that off-the-shelf PL is a feasible substitute for fetal bovine serum in MSC cultures. © 2017 AABB.
Störmer, Melanie; Kleesiek, Knut; Dreier, Jens
2007-01-01
Nucleic acid isolation, the most technically demanding and laborious procedure performed in molecular diagnostics, harbors the potential for improvements in automation. A recent development is the use of magnetic beads covered with nucleic acid-binding matrices. We adapted this technology with a broad-range 23S rRNA real-time reverse transcription (RT)-PCR assay for fast and sensitive detection of bacterial contamination of blood products. We investigated different protocols for an automated high-volume extraction method based on magnetic-separation technology for the extraction of bacterial nucleic acids from platelet concentrates (PCs). We added 2 model bacteria, Staphylococcus epidermidis and Escherichia coli, to a single pool of apheresis-derived, single-donor platelets and assayed the PCs by real-time RT-PCR analysis with an improved primer-probe system and locked nucleic acid technology. Co-amplification of human beta(2)-microglobulin mRNA served as an internal control (IC). We used probit analysis to calculate the minimum concentration of bacteria that would be detected with 95% confidence. For automated magnetic bead-based extraction technology with the real-time RT-PCR, the 95% detection limit was 29 x 10(3) colony-forming units (CFU)/L for S. epidermidis and 22 x 10(3) CFU/L for E. coli. No false-positive results occurred, either due to nucleic acid contamination of reagents or externally during testing of 1030 PCs. High-volume nucleic acid extraction improved the detection limit of the assay. The improvement of the primer-probe system and the integration of an IC make the RT-PCR assay appropriate for bacteria screening of platelets.
Borja, Miguel; Galan, Jacob Anthony; Cantu, Esteban; Zugasti-Cruz, Alejandro; Rodríguez-Acosta, Alexis; Lazcano, David; Lucena, Sara; Suntravat, Montamas; Sánchez, y Elda Eliza
2016-01-01
The Tamaulipan rock rattlesnake (Crotalus lepidus morulus) is a montane snake that occurs in the humid pine-oak forest and the upper cloud forest of the Sierra Madre Oriental in southwestern Tamaulipas, central Nuevo Leon, and southeastern Coahuila in Mexico. Venom from this rattlesnake was fractionated by High-Performance Liquid Chromatography for the purpose of discovering disintegrin molecules. Disintegrins are non-enzymatic, small molecular weight peptides that interfere with cell-cell and cell-matrix interactions by binding to various cell receptors. Eleven fractions were collected by anion exchange chromatography and pooled into six groups (I, II, III, IV, V, and VI). Proteins of the six groups were analyzed by SDS-PAGE and western blot using antibodies raised against a disintegrin. The antibodies recognized different protein bands in five (II, III, IV, V, and VI) of six groups in a molecular mass range of 7 to 105 kDa. Western blot analysis revealed fewer protein bands in the higher molecular mass range and two bands in the disintegrin weight range in group II compared with the other four groups. Proteins in group II were further separated into nine fractions using reverse phase C18 chromatography. Fraction 4 inhibited platelet aggregation and was named morulustatin, which exhibited a single band with a molecular mass of approximately 7 kDa. Mass spectrometry analysis of fraction 4 revealed the identification of disintegrin peptides LRPGAQCADGLCCDQCR (MH+ 2035.84) and AGEECDCGSPANCCDAATCK (MH+ 2328.82). Morulustatin inhibited ADP-induced platelet aggregation in human whole blood and was concentration-dependent with an IC50 of 89.5 nM ± 12. PMID:28713196
Annual report, FY 1979 Spent fuel and fuel pool component integrity.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Johnson, A.B. Jr.; Bailey, W.J.; Schreiber, R.E.
International meetings under the BEFAST program and under INFCE Working Group No. 6 during 1978 and 1979 continue to indicate that no cases of fuel cladding degradation have developed on pool-stored fuel from water reactors. A section from a spent fuel rack stand, exposed for 1.5 y in the Yankee Rowe (PWR) pool had 0.001- to 0.003-in.-deep (25- to 75-..mu..m) intergranular corrosion in weld heat-affected zones but no evidence of stress corrosion cracking. A section of a 304 stainless steel spent fuel storage rack exposed 6.67 y in the Point Beach reactor (PWR) spent fuel pool showed no significant corrosion.more » A section of 304 stainless steel 8-in.-dia pipe from the Three Mile Island No. 1 (PWR) spent fuel pool heat exchanger plumbing developed a through-wall crack. The crack was intergranular, initiating from the inside surface in a weld heat-affected zone. The zone where the crack occurred was severely sensitized during field welding. The Kraftwerk Union (Erlangen, GFR) disassembled a stainless-steel fuel-handling machine that operated for 12 y in a PWR (boric acid) spent fuel pool. There was no evidence of deterioration, and the fuel-handling machine was reassembled for further use. A spent fuel pool at a Swedish PWR was decontaminated. The procedure is outlined in this report.« less
Analysis of Terrestrial Carbon Stocks in a Small Catchment of Northeastern Siberia
NASA Astrophysics Data System (ADS)
Heard, K.; Natali, S.; Bunn, A. G.; Loranty, M. M.; Kholodov, A. L.; Schade, J. D.; Berner, L. T.; Spektor, V.; Zimov, N.; Alexander, H. D.
2015-12-01
As arctic terrestrial ecosystems comprise about one-third of the global terrestrial ecosystem carbon total, understanding arctic carbon cycling and the feedback of terrestrial carbon pools to accelerated warming is an issue of global concern. For this research, we examined above- and belowground carbon stocks in a larch-dominated catchment underlain by yedoma and located within the Kolyma River watershed in northeastern Siberia. We quantified carbon stocks in vegetation, active layer, and permafrost, and we assessed the correlation between plant and active layer carbon pools and four environmental correlates — slope, solar insolation, canopy density, and leaf area index — at 20 sites. Carbon in the active layer was approximately four times greater than aboveground carbon pools (972 g C m-2), and belowground carbon to 1 m depth was approximately 18 times greater than aboveground carbon pools. Canopy density and slope had a robust positive association with aboveground carbon pools, and soil moisture was positively related to %C in organic, thawed mineral and permafrost soil. Thaw depth was negatively correlated with moss cover and larch biomass, highlighting the importance of vegetation and surface characteristics on permafrost carbon vulnerability. These data suggest that landscape and ecosystem characteristics affect carbon accumulation and storage, but they also play an important role in stabilizing permafrost carbon pools.
Possible effect of biotechnology on plant gene pools in Turkey.
Demir, Aynur
2015-01-02
The recent rapid developments in biotechnology have made great contributions to the study of plant gene pools. The application of in vitro methods in freeze storage and DNA protection techniques in fast production studies has made major advances. From that aspect, biotechnology is an indispensable means for the protection of plant gene pools, which includes the insurance of sustainable agriculture and development of species. Besides all the positive developments, one of the primary risks posed by the uncontrolled spreading of genetically modified organisms is the possibility for other non-target organisms to be negatively affected. Genes of plant origin should be given priority in this type of studies by taking into consideration such negative effects that may result in disruption of ecological balance and damage to plant genetic pools. Turkey, due to its ecological conditions and history, has a very important position in terms of plant gene pools. This richness ought to be protected without corrupting its natural quality and natural evolution process in order to provide the sources of species that will be required for future sustainable agricultural applications. Thus, attention should be paid to the use of biotechnological methods, which play an important role especially in the protection and use of local and original plant gene pools.
Possible effect of biotechnology on plant gene pools in Turkey
Demir, Aynur
2015-01-01
The recent rapid developments in biotechnology have made great contributions to the study of plant gene pools. The application of in vitro methods in freeze storage and DNA protection techniques in fast production studies has made major advances. From that aspect, biotechnology is an indispensable means for the protection of plant gene pools, which includes the insurance of sustainable agriculture and development of species. Besides all the positive developments, one of the primary risks posed by the uncontrolled spreading of genetically modified organisms is the possibility for other non-target organisms to be negatively affected. Genes of plant origin should be given priority in this type of studies by taking into consideration such negative effects that may result in disruption of ecological balance and damage to plant genetic pools. Turkey, due to its ecological conditions and history, has a very important position in terms of plant gene pools. This richness ought to be protected without corrupting its natural quality and natural evolution process in order to provide the sources of species that will be required for future sustainable agricultural applications. Thus, attention should be paid to the use of biotechnological methods, which play an important role especially in the protection and use of local and original plant gene pools. PMID:26019612
Watanabe, Naohide; Nogawa, Masayuki; Ishiguro, Mariko; Maruyama, Hitomi; Shiba, Masayuki; Satake, Masahiro; Eto, Koji; Handa, Makoto
2017-08-01
To bridge the gap between in vitro function and clinical efficacy of platelet (PLT) transfusion products, reliable in vivo PLT functional assays for hemostasis and survival in animal models are required. However, there are no standardized methods for assessing the in vivo quality of transfused human PLTs. Plasma-depleted human PLT concentrates (PCs; Day 3, Day 5, Day 7, Day 10, and damaged) were transfused into busulfan-induced rabbits with thrombocytopenia with prolonged bleeding times 1 day after treatment with ethyl palmitate (EP) to block their reticuloendothelial systems. The hemostatic effect of PC transfusion was evaluated by the ear fine vein bleeding time. For the in vivo survival assay, splenectomized EP-treated rabbits were transfused with human PCs, and viability of the human PLTs in the rabbits was determined by flow cytometry using human PLT-specific antibodies and Trucount tubes. The hemostatic effect of PCs was slightly reduced with increasing storage periods for early time points, but more dramatically reduced for later time points. PLT survival was similar after 3 and 7 days of storage, but PLTs stored for 10 days showed significantly poorer survival than those stored only 3 days. Our new and improved protocol for in vivo assessment of transfused PLTs is sufficiently sensitive to detect subtle changes in hemostatic function and viability of human PLTs transfused into rabbit models. This protocol could contribute to preclinical in vivo functional assessment and clinical quality assurance of emerging novel PLT products such as cultured cell-derived human PLTs. © 2017 AABB.
NASA Astrophysics Data System (ADS)
Elosegi, Arturo; Díez, José Ramón; Flores, Lorea; Molinero, Jon
2017-02-01
Large wood (LW, or pieces of dead wood longer than 1 m and thicker than 10 cm in diameter) is a key element in forested streams, but its abundance has decreased worldwide as a result of snagging and clearing of riparian forests. Therefore, many restoration projects introduce LW into stream channels to enhance geomorphology, biotic communities, and ecosystem functioning. Because LW enhances the retention of organic matter and sediments, its restoration can reduce siltation in receiving reservoirs, although so far little information on this subject is available. We studied the effects of restoring the natural loading of LW in four streams in the Aiako Harria Natural Park (the Basque Country, Spain) in pool abundance, channel form, and storage of organic matter and sediments. In all reaches log jams induced the formation of new geomorphic features and changes in physical habitat, especially an increase in the number and size of pools and in the formation of gravel bars and organic deposits. The storage of organic matter increased 5- to 88-fold and streambed level rose 7 ± 4 to 21 ± 4 cm on average, resulting in the storage of 35.2 ± 19.7 to 711 ± 375 m3 (733-1400 m3 ha- 1 y- 1) of sediment per reach. Extrapolation of these results to the entire drainage basin suggests that basinwide restoration of LW loading would enhance the retention potential of stream channels by 66,817 ± 27,804 m3 (1075 m3 ha- 1 y- 1) of sediment and by 361 t (5.32 T ha- 1 y- 1) of organic matter, which represents 60% of the estimated annual inputs of sediments to the downstream Añarbe Reservoir and almost twice as much as the annual input of organic matter to the entire river network. Therefore, basinwide restoration of LW loading is a potentially important tool to manage catchments that feed reservoirs, where retention of sediments and organic matter can be considered important ecosystem services as they reduce reservoir siltation.
Dou, Xiaolin; He, Ping; Cheng, Xiaoli; Zhou, Wei
2016-01-01
Quantification of dynamics of soil organic carbon (SOC) pools under the influence of long-term fertilization is essential for predicting carbon (C) sequestration. We combined soil chemical fractionation with stable C isotope analyses to investigate the C dynamics of the various SOC pools after 25 years of fertilization. Five types of soil samples (0–20, 20–40 cm) including the initial level (CK) and four fertilization treatments (inorganic nitrogen fertilizer, IN; balanced inorganic fertilizer, NPK; inorganic fertilizer plus farmyard manure, MNPK; inorganic fertilizer plus corn straw residue, SNPK) were separated into recalcitrant and labile fractions, and the fractions were analysed for C content, C:N ratios, δ13C values, soil C and N recalcitrance indexes (RIC and RIN). Chemical fractionation showed long-term MNPK fertilization strongly increased the SOC storage in both soil layers (0–20 cm = 1492.4 gC m2 and 20–40 cm = 1770.6 gC m2) because of enhanced recalcitrant C (RC) and labile C (LC). The 25 years of inorganic fertilizer treatment did not increase the SOC storage mainly because of the offsetting effects of enhanced RC and decreased LC, whereas no clear SOC increases under the SNPK fertilization resulted from the fast decay rates of soil C. PMID:26750143
Differences in sedge fen vegetation upstream and downstream from a managed impoundment
Kowalski, Kurt P.; Wilcox, Douglas A.
2003-01-01
The U.S. Fish and Wildlife Service proposed the restoration of wetlands impacted by a series of drainage ditches and pools located in an extensive undeveloped peatland in the Seney National Wildlife Refuge, Michigan. This study examined the nature and extent of degradation to the Marsh Creek wetlands caused by alteration of natural hydrology by a water-storage pool (C-3 Pool) that intersects the Marsh Creek channel. We tested the hypothesis that a reduction in moderate-intensity disturbance associated with natural water-level fluctuations below the C-3 dike contributed to lower species richness, reduced floristic quality and a larger tree and shrub component than vegetation upstream from the pool. Wetland plant communities were sampled quantitatively and analyzed for species richness, floristic quality and physiognomy. Aerial photographs, GIS databases and GPS data contributed to the characterization and analysis of the Marsh Creek wetlands. Results showed that there was lower species richness in vegetated areas downstream from the pool, but not the anticipated growth in shrubs. Wetland vegetation upstream and downstream from the pool had similar floristic quality, except for a greater number of weedy taxa above the pool. Seepage through the pool dike and localized ground-water discharge created conditions very similar to those observed around beaver dams in Marsh Creek. In essence, the dike containing the C-3 Pool affected hydrology and wetland plant communities in a manner similar to an enormous beaver dam, except that it did not allow seasonal flooding episodes to occur. Management actions to release water from the pool into the original Marsh Creek channel at certain times and in certain amounts that mimic the natural flow regime would be expected to promote greater plant species richness and minimize the negative impacts of the dike.
78 FR 38308 - PK Ventures, Inc.; North Carolina; Notice Soliciting Applications
Federal Register 2010, 2011, 2012, 2013, 2014
2013-06-26
... DEPARTMENT OF ENERGY Federal Energy Regulatory Commission [Project No. 4093-031] PK Ventures, Inc... at normal pool elevation of 315 feet mean sea level and a gross storage capacity of 100 acre-feet; and (5) appurtenant facilities. The project operates run-of-river and generates and estimated average...
Carbon sequestration in harvested wood products
K. Skog
2013-01-01
Carbon is continuously cycled among these storage pools and between forest ecosystems and the atmosphere as a result of biological processes in forests (e.g., photosynthesis, respiration, growth, mortality, decomposition, and disturbances such as fires or pest outbreaks) and anthropogenic activities (e.g., harvesting, thinning, clearing, and replanting). As trees...
10 CFR 36.53 - Operating and emergency procedures.
Code of Federal Regulations, 2012 CFR
2012-01-01
... 10 Energy 1 2012-01-01 2012-01-01 false Operating and emergency procedures. 36.53 Section 36.53 Energy NUCLEAR REGULATORY COMMISSION LICENSES AND RADIATION SAFETY REQUIREMENTS FOR IRRADIATORS Operation... the source storage pool; (6) A prolonged loss of electrical power; (7) A fire alarm or explosion in...
10 CFR 36.53 - Operating and emergency procedures.
Code of Federal Regulations, 2014 CFR
2014-01-01
... 10 Energy 1 2014-01-01 2014-01-01 false Operating and emergency procedures. 36.53 Section 36.53 Energy NUCLEAR REGULATORY COMMISSION LICENSES AND RADIATION SAFETY REQUIREMENTS FOR IRRADIATORS Operation... the source storage pool; (6) A prolonged loss of electrical power; (7) A fire alarm or explosion in...
10 CFR 36.53 - Operating and emergency procedures.
Code of Federal Regulations, 2013 CFR
2013-01-01
... 10 Energy 1 2013-01-01 2013-01-01 false Operating and emergency procedures. 36.53 Section 36.53 Energy NUCLEAR REGULATORY COMMISSION LICENSES AND RADIATION SAFETY REQUIREMENTS FOR IRRADIATORS Operation... the source storage pool; (6) A prolonged loss of electrical power; (7) A fire alarm or explosion in...
Ecohydrological coupling at the watershed scale is poorly characterized. While soil-water storage is dynamic and strongly influenced by plants, few integrated tools exist for quantifying the spatial and temporal dynamics and interactions among the major components of the terrestr...
Code of Federal Regulations, 2011 CFR
2011-04-01
... occupied had it been in operation. (3) A proposed annual rule of operation for the storage reservoir or... the reservoir or reservoirs showing the maximum, average, and minimum operating pool levels, the..., weekly and daily, during periods of low and normal flows after the plant is in operation and the system...
Serrano, Katherine; Levin, Elena; Culibrk, Brankica; Weiss, Sandra; Scammell, Ken; Boecker, Wolfgang F; Devine, Dana V
2010-01-01
BACKGROUND In high-volume processing environments, manual breakage of in-line closures can result in repetitive strain injury (RSI). Furthermore, these closures may be incorrectly opened causing shear-induced hemolysis. To overcome the variability of in-line closure use and minimize RSI, Fresenius Kabi developed a new in-line closure, the CompoFlow, with mechanical openers. STUDY DESIGN AND METHODS The consistency of the performance of the CompoFlow closure device was assessed, as was its effect on component quality. A total of 188 RBC units using CompoFlow blood bag systems and 43 using the standard bag systems were produced using the buffy coat manufacturing method. Twenty-six CompoFlow platelet (PLT) concentrates and 10 control concentrates were prepared from pools of four buffy coats. RBCs were assessed on Days 1, 21, and 42 for cellular variables and hemolysis. PLTs were assessed on Days 1, 3, and 7 for morphology, CD62P expression, glucose, lactate, and pH. A total of 308 closures were excised after processing and the apertures were measured using digital image analysis. RESULTS The use of the CompoFlow device significantly improved the mean extraction time with 0.46 ± 0.11 sec/mL for the CompoFlow units and 0.52 ± 0.13 sec/mL for the control units. The CompoFlow closures showed a highly reproducible aperture after opening (coefficient of variation, 15%) and the device always remained opened. PLT and RBC products showed acceptable storage variables with no differences between CompoFlow and control. CONCLUSIONS The CompoFlow closure devices improved the level of process control and processing time of blood component production with no negative effects on product quality. PMID:20529007
Naskou, Maria C; Sumner, Scarlett M; Chocallo, Anna; Kemelmakher, Hannah; Thoresen, Merrilee; Copland, Ian; Galipeau, Jacques; Peroni, John F
2018-03-22
Mesenchymal stem cells (MSCs) produced for clinical purposes rely on culture media containing fetal bovine serum (FBS) which is xenogeneic and has the potential to significantly alter the MSC phenotype, rendering these cells immunogenic. As a result of bovine-derived exogenous proteins expressed on the cell surface, MSCs may be recognized by the host immune system as non-self and be rejected. Platelet lysate (PL) may obviate some of these concerns and shows promising results in human medicine as a possible alternative to FBS. Our goal was to evaluate the use of equine platelet lysate (ePL) pooled from donor horses in place of FBS to culture equine MSCs. We hypothesized that ePL, produced following apheresis, will function as the sole media supplement to accelerate the expansion of equine bone marrow-derived MSCs without altering their phenotype and their immunomodulatory capacity. Platelet concentrate was obtained via plateletpheresis and ePL were produced via freeze-thaw and centrifugation cycles. Population doublings (PD) and doubling time (DT) of bone marrow-derived MSCs (n = 3) cultured with FBS or ePL media were calculated. Cell viability, immunophenotypic analysis, and trilineage differentiation capacity of MSCs were assessed accordingly. To assess the ability of MSCs to modulate inflammatory responses, E. coli lipopolysaccharide (LPS)-stimulated monocytes were cocultured with MSCs cultured in the two different media formulations, and cell culture supernatants were assayed for the production of tumor necrosis factor (TNF)-α. Our results showed that MSCs cultured in ePL media exhibited similar proliferation rates (PD and DT) compared with those cultured in FBS at individual time points. MSCs cultured in ePL showed a statistically significant increased viability following a single washing step, expressed similar levels of MSC markers compared to FBS, and were able to differentiate towards the three lineages. Finally, MSCs cultured in ePL efficiently suppressed the release of TNF-α when exposed to LPS-stimulated monocytes similar to those cultured in FBS. ePL has the potential to be used for the expansion of MSCs before clinical application, avoiding the concerns associated with the use of FBS.
Little, C L; Rhoades, J R; Hucklesby, L; Greenwood, M; Surman-Lee, S; Bolton, F J; Meldrum, R; Wilson, I; McDonald, C; de Pinna, E; Threlfall, E J; Chan, C H
2008-01-01
This survey was launched after an unusual number of Salmonella Enteritidis outbreaks associated with the use of eggs in food service premises in England and Wales. Between November 2005 and December 2006, 9,528 eggs (1,588 pooled samples of 6 eggs) were collected from 1,567 food service premises in the United Kingdom, most of which (89%) were produced in the United Kingdom. Salmonella was isolated from 6 (0.38%) pools of eggs. Of these, 5 (0.31%) were Salmonella Enteritidis, which were further characterized to phage types (PTs): PT 4 (0.19%), PT 8 (0.06%), and PT 12 (0.06%). Salmonella Mbandaka was also isolated (0.06%). Salmonella was detected from five and one of pooled eggs samples that were produced in the United Kingdom and Germany, respectively; these were from different producers. The study showed evidence of poor egg storage and handling practices in food service premises, in that 55% did not store eggs under refrigerated conditions; 20.7% of eggs had expired "best before" dates or were in use after 3 weeks of lay, indicating poor stock rotation; and 37.1% pooled eggs not intended for immediate service. Eggs are a commonly consumed food that may occasionally be contaminated with Salmonella at different rates, according to their country of origin. The food service sector needs to be aware of this continuing hazard, receive appropriate food safety and hygiene training on storage and usage of raw shell eggs, adopt appropriate control measures, and follow advice provided by national food agencies in order to reduce the risk of infection.
Hyperforin changes the zinc-storage capacities of brain cells.
Gibon, Julien; Richaud, Pierre; Bouron, Alexandre
2011-12-01
In vitro and in vivo experiments were carried out to investigate the consequences on brain cells of a chronic treatment with hyperforin, a plant extract known to dissipate the mitochondrial membrane potential and to release Zn(2+) and Ca(2+) from these organelles. Dissociated cortical neurons were grown in a culture medium supplemented with 1 μM hyperforin. Live-cell imaging experiments with the fluorescent probes FluoZin-3 and Fluo-4 show that a 3 day-hyperforin treatment diminishes the size of the hyperforin-sensitive pools of Ca(2+) and Zn(2+) whereas it increases the size of the DTDP-sensitive pool of Zn(2+) without affecting the ionomycin-sensitive pool of Ca(2+). When assayed by quantitative PCR the levels of mRNA coding for metallothioneins (MTs) I, II and III were increased in cortical neurons after a 3 day-hyperforin treatment. This was prevented by the zinc chelator TPEN, indicating that the plant extract controls the expression of MTs in a zinc-dependent manner. Brains of adult mice who received a daily injection (i.p.) of hyperforin (4 mg/kg/day) for 4 weeks had a higher sulphur content than control animals. They also exhibited an enhanced expression of the genes coding for MTs. However, the long-term treatment did not affect the brain levels of calcium and zinc. Based on these results showing that hyperforin influences the size of the internal pools of Zn(2+), the expression of MTs and the brain cellular sulphur content, it is proposed that hyperforin changes the Zn-storage capacity of brain cells and interferes with their thiol status. Copyright © 2011 Elsevier Ltd. All rights reserved.
Effect of tree line advance on carbon storage in NW Alaska
Wilmking, M.; Harden, J.; Tape, K.
2006-01-01
We investigated the size, distribution, and temporal dynamics of ecosystem carbon (C) pools in an area of recent tree line advance, northwest Alaska. Repeat aerial photographs show forest cover increased ???10% in our study area since 1949. We sampled C pools of four principal ecosystem types, tussock tundra, shrub tundra, woodland, and forest, all located on a 600-800 year old river terrace. Significant differences between ecosystem C pools, both above ground and below ground existed. Tundra sites store >22.2 kg C/m2, shrub tundra sites and woodland sites store 9.7 kg C/m2 and 14.3 kg C/m2, respectively, and forest sites store 14.4 kg C/m2. Landscape variation of total ecosystem C was primarily due to organic soil C and was secondarily due to C stored in trees. Soil C/N profiles of shrub tundra sites and woodland sites showed similarities with forest site soils at surface and tundra site soils at depth. We hypothesize that tundra systems transformed to forest systems in this area under a progression of permafrost degradation and enhanced drainage. On the basis of C pool estimates for the different ecosystem types, conversion of tundra sites to forest may have resulted in a net loss of > 7.8 kg C/m2, since aboveground C gains were more than offset by belowground C losses to decomposition in the tundra sites. Tree line advance therefore might not increase C storage in high-latitude ecosystems and thus might not, as previously suggested, act as a negative feedback to warming. Key to this hypothesis and to its projection to future climate response is the fate of soil carbon upon warming and permafrost drainage. Copyright 2006 by the American Geophysical Union.
Transfusions of blood products and cancer outcomes.
Velásquez, J F; Cata, J P
2015-10-01
Approximately half of cancer patients scheduled for major surgery are anemic. Also, a significant number of patients will present to the operating room with low platelet counts and coagulopathic disorders. Unfortunately, administration of red blood cells, platelets concentrates and fresh-frozen plasma is associated with unwanted adverse effects including fever, hemolytic reactions and transfusion-related immunomodulation (TRIM). TRIM is a multifactorial immunologic phenomenon in the recipient mediated by donor leukocytes, microparticles such as ectosomes, and growth factors. As some of these molecules are secreted in a time-dependent manner, blood storage time may play an important in TRIM, although the evidence is limited. Perioperative administration of red blood cells and associated TRIM has also been associated with increased recurrence of certain solid tumors, such as colorectal, lung, and hepatobiliary tumors. In this continuing education article, we review the available evidence on how perioperative blood product transfusions can affect oncological outcomes, such as cancer recurrence. Copyright © 2014 Sociedad Española de Anestesiología, Reanimación y Terapéutica del Dolor. Publicado por Elsevier España, S.L.U. All rights reserved.
NASA Astrophysics Data System (ADS)
Tonitto, C.; Goodale, C. L.; Ollinger, S. V.; Jenkins, J.
2009-12-01
Anthropogenic forcing of the C and N cycles has caused rapid change in atmospheric CO2 and N deposition, with complex and uncertain effects on forest C and N balance. We developed the PnET-SOM model to enhance the model description of carbon and nitrogen coupling. Here we applied PnET-SOM to study changes to ecosystem carbon storage across a nitrogen deposition gradient. We designed the PnET-SOM model to: 1) represent SOM structured around measurable SOM pools, 2) expand simulated soil horizon complexity beyond the 1-box approach to hydrology and SOM structure used in PnET-CN, 3) model humified and mineral associated SOM using parameters derived from C14 field studies, and 4) couple C and N cycles to allow N-limitation of decomposition and plant growth. We explicitly modeled labile, biochemically recalcitrant (humified SOM), and physically-chemically protected (mineral associated SOM) C pools. These SOM pools are modeled in distinct soil horizons including: a forest floor, a mixed organic horizon, an A horizon, and a B horizon. Slow turnover pools of the A and B horizon constitute a significant proportion of SOC; explicitly modeling a deeper soil profile is important for estimating ecosystem SOC storage. In the latest version of PnET-SOM, we described N mineralization-immobilization in the forest floor based on equations derived in the LIDET synthesis study. Validation of the PnET-SOM model was conducted using 1) long-term water flux and nitrate leaching data from the Hubbard Brook LTER, 2) CO2 respiration observations from the Harvard Forest LTER, and 3) C and N stock and flux observations from the Harvard Forest LTER. In this work, we applied the PnET-SOM model to study the effects of an N deposition gradient on SOC dynamics over a 300 year simulation. We represented the effects of N deposition on litter decomposition by varying the exponential decay parameters of the litter layer based on observations from the Harvard Forest N addition experiment. We derived the change in limit value across the N deposition gradient simulated by applying a polynomial fit to limit values observed in the Harvard Forest N addition experimental plots. In PnET-SOM, changes in SOC dynamics under varying N deposition rates are derived from 1) change in litter decomposition rate, which directly affects OM inputs into soil pools, and 2) altered N availability which limits decomposition of OM throughout the soil profile. In our model application, the coarse woody debris pool decreased with increasing N deposition, while the forest floor pools increased. Relative to SOC pools under current N deposition rates at the Harvard Forest (8 gN/m2), deposition at a rate of 20 gN/m2 over a 300 year simulation resulted in a 3.4% increase in the O horizon humified SOC pool, a 2.3% increase in the A horizon humified SOC pool, a 1.3% increase in the A horizon mineral associated SOC pool, and a 0.14% increase in the B horizon mineral associated SOC pool.
Ambrosio, Andrea L.; Boyle, Judith A.; Di Pietro, Santiago M.
2015-01-01
Platelet dense granules (PDGs) are acidic calcium stores essential for normal hemostasis. They develop from late endosomal compartments upon receiving PDG-specific proteins through vesicular trafficking, but their maturation process is not well understood. Here we show that two-pore channel 2 (TPC2) is a component of the PDG membrane that regulates PDG luminal pH and the pool of releasable Ca2+. Using a genetically encoded Ca2+ biosensor and a pore mutant TPC2, we establish the function of TPC2 in Ca2+ release from PDGs and the formation of perigranular Ca2+ nanodomains. For the first time, Ca2+ spikes around PDGs—or any organelle of the endolysosome family—are visualized in real time and revealed to precisely mark organelle “kiss-and-run” events. Further, the presence of membranous tubules transiently connecting PDGs is revealed and shown to be dramatically enhanced by TPC2 in a mechanism that requires ion flux through TPC2. “Kiss-and-run” events and tubule connections mediate transfer of membrane proteins and luminal content between PDGs. The results show that PDGs use previously unknown mechanisms of membrane dynamics and content exchange that are regulated by TPC2. PMID:26202466
Hussain, Nasir; Johal, Herman; Bhandari, Mohit
2017-01-01
Within orthopedics, the use of platelet-rich plasma (PRP) has been rapidly increasing in popularity, however, its true effectiveness has yet to be fully established. Several studies find that injecting PRP to the site of injury does not provide any significant benefit with respect to clinical outcomes; however, many others report the contrary. Due to the conflicting evidence and multiple meta-analyses conducted on the topic, a literature review of high-quality evidence on the use of PRP for common orthopaedic conditions was performed. Thus far, the evidence appears to suggest that PRP may provide some benefit in patients who present with knee osteoarthritis or lateral epicondylitis. On the other hand, evidence appears to be inconsistent or shows a minimal benefit for PRP usage in rotator cuff repair, patellar and Achilles tendinopathies, hamstring injuries, anterior cruciate ligament (ACL) repair, and medial epicondylitis. There is limited confidence in the conclusions from the published meta-analyses due to issues with statistical pooling, and limited subgroup analyses exploring the substantial heterogeneity across studies. Evidence-based clinicians considering the use of PRP in their patients with musculoskeletal injuries should be weary that the literature appears to be inconsistent and thus far, inconclusive. PMID:28990574
Radu, Claudia M; Spiezia, Luca; Bulato, Cristiana; Gavasso, Sabrina; Campello, Elena; Sartorello, Francesca; Castoldi, Elisabetta; Simioni, Paolo
2016-11-01
Although human megakaryocytes can synthesize factor V (FV), platelet FV derives largely from endocytosis of plasma FV. Recently, it has been shown that plasma transfusions can replenish the platelet FV pool in parahaemophilic patients. Here we corroborate this finding by showing FV endocytosis by ex vivo differentiated megakaryocytes derived from patients with inherited parahaemophilia. Mononuclear stem cells isolated from peripheral blood of healthy subjects and of three patients with severe parahaemophilia were cultured in the presence of thrombopoietin and interleukin-3 and differentiated into CD41-positive polynucleated megakaryocytes. Exogenous purified FV was added to the culture medium to evaluate FV endocytosis. Immunofluorescence staining revealed abundant FV expression in megakaryocytes derived from healthy donors, but no FV expression in those derived from patients with severe parahaemophilia. However, after the addition of purified FV to the culture medium, megakaryocytes from parahaemophilia patients became positive upon FV immunostaining, suggesting endocytosis of exogenous FV. Endocytosed FV retained factor Xa-co-factor activity as assessed by a prothrombin time-based functional test in megakaryocyte lysates. Addition of exogenous FV to culture medium can restore the FV content of megakaryocytes derived from patients with severe FV defects. This rescue mechanism can have important clinical implications in the management of parahaemophilia patients. © 2016 John Wiley & Sons Ltd.
Szymusiak, Magdalena; Donovan, Alexander J; Smith, Stephanie A; Ransom, Ross; Shen, Hao; Kalkowski, Joseph; Morrissey, James H; Liu, Ying
2016-01-20
Platelet-sized polyphosphate (polyP) was functionalized on the surface of gold nanoparticles (GNPs) via a facile conjugation scheme entailing EDAC (N-(3-(dimethylamino)propyl)-N'-ethylcarbodiimide hydrochloride)-catalyzed phosphoramidation of the terminal phosphate of polyP to cystamine. Subsequent reduction of the disulfide moiety allowed for anchoring to the colloidal surface. The ability of the synthesized polyP-GNPs to initiate the contact pathway of clotting in human pooled normal plasma (PNP) was then assayed by quantifying changes in viscous, mechanical, and optical properties upon coagulation. It is revealed that the polyP-GNPs are markedly superior contact activators compared to molecularly dissolved, platelet-sized polyP (of equivalent polymer chain length). Moreover, the particles' capacity to mobilize Factor XII (FXII) and its coactivating proteins appear to be identical to very-long-chain polyP typically found in bacteria. These data imply that nanolocalization of anionic procoagulants on colloidal surfaces, achieved through covalent anchoring, may yield a robust contact surface with the ability to sufficiently cluster active clotting factors together above their threshold concentrations to cease bleeding. The polyP-GNPs therefore serve as a promising foundation in the development of a nanoparticle hemostat to treat a range of hemorrhagic scenarios.
The Design and Testing of a Dual Fiber Textile Matrix for Accelerating Surface Hemostasis
Fischer, Thomas H.; Vournakis, John N.; Manning, James E.; McCurdy, Shane L.; Rich, Preston B.; Nichols, Timothy C.; Scull, Christopher M.; McCord, Marian G.; Decorta, Joseph A.; Johnson, Peter C.; Smith, Carr J.
2011-01-01
The standard treatment for severe traumatic injury is frequently compression and application of gauze dressing to the site of hemorrhage. However, while able to rapidly absorb pools of shed blood, gauze fails to provide strong surface (topical) hemostasis. The result can be excess hemorrhage-related morbidity and mortality. We hypothesized that cost-effective materials (based on widespread availability of bulk fibers for other commercial uses) could be designed based on fundamental hemostatic principles to partially emulate the wicking properties of gauze while concurrently stimulating superior hemostasis. A panel of readily available textile fibers was screened for the ability to activate platelets and the intrinsic coagulation cascade in vitro. Type E continuous filament glass and a specialty rayon fiber were identified from the material panel as accelerators of hemostatic reactions and were custom woven to produce a dual fiber textile bandage. The glass component strongly activated platelets while the specialty rayon agglutinated red blood cells. In comparison with gauze in vitro, the dual fiber textile significantly enhanced the rate of thrombin generation, clot generation as measured by thromboelastography, adhesive protein adsorption and cellular attachment and activation. These results indicate that hemostatic textiles can be designed that mimic gauze in form but surpass gauze in ability to accelerate hemostatic reactions. PMID:19489008
McHale, Melissa R; Hall, Sharon J; Majumdar, Anandamayee; Grimm, Nancy B
2017-03-01
Human modification and management of urban landscapes drastically alters vegetation and soils, thereby altering carbon (C) storage and rates of net primary productivity (NPP). Complex social and ecological processes drive vegetation cover in cities, leading to heterogeneity in C dynamics depending on regional climate, land use, and land cover. Recent work has demonstrated homogenization in ecological processes within human-dominated landscapes (the urban convergence hypothesis) in soils and biotic communities. However, a lack of information on vegetation in arid land cities has hindered an understanding of potential C storage and NPP convergence across a diversity of ecosystem types. We estimated C storage and NPP of trees and shrubs for six different land-use types in the arid metropolis of Phoenix, Arizona, USA, and compared those results to native desert ecosystems, as well as other urban and natural systems around the world. Results from Phoenix do not support the convergence hypothesis. In particular, C storage in urban trees and shrubs was 42% of that found in desert vegetation, while NPP was only 20% of the total NPP estimated for comparable natural ecosystems. Furthermore, the overall estimates of C storage and NPP associated with urban trees in the CAP ecosystem were much lower (8-63%) than the other cities included in this analysis. We also found that C storage (175.25-388.94 g/m 2 ) and NPP (8.07-15.99 g·m -2 ·yr -1 ) were dominated by trees in the urban residential land uses, while in the desert, shrubs were the primary source for pools (183.65 g/m 2 ) and fluxes (6.51 g·m -2 ·yr -1 ). These results indicate a trade-off between shrubs and trees in arid ecosystems, with shrubs playing a major role in overall C storage and NPP in deserts and trees serving as the dominant C pool in cities. Our research supports current literature that calls for the development of spatially explicit and standardized methods for analyzing C dynamics associated with vegetation in urbanizing areas. © 2016 by the Ecological Society of America.
Surfactants reduce platelet-bubble and platelet-platelet binding induced by in vitro air embolism.
Eckmann, David M; Armstead, Stephen C; Mardini, Feras
2005-12-01
The effect of gas bubbles on platelet behavior is poorly characterized. The authors assessed platelet-bubble and platelet-platelet binding in platelet-rich plasma in the presence and absence of bubbles and three surface-active compounds. Platelet-rich plasma was prepared from blood drawn from 16 volunteers. Experimental groups were surfactant alone, sparging (microbubble embolization) alone, sparging with surfactant, and neither sparging nor surfactant. The surfactants were Pluronic F-127 (Molecular Probes, Eugene, OR), Perftoran (OJSC SPC Perftoran, Moscow, Russia), and Dow Corning Antifoam 1510US (Dow Corning, Midland, MI). Videomicroscopy images of specimens drawn through rectangular glass microcapillaries on an inverted microscope and Coulter counter measurements were used to assess platelet-bubble and platelet-platelet binding, respectively, in calcium-free and recalcified samples. Histamine-induced and adenosine diphosphate-induced platelet-platelet binding were measured in unsparged samples. Differences between groups were considered significant for P < 0.05 using analysis of variance and the Bonferroni correction. Sixty to 100 platelets adhered to bubbles in sparged, surfactant-free samples. With sparging and surfactant, few platelets adhered to bubbles. Numbers of platelet singlets and multimers not adherent to bubbles were different (P < 0.05) compared both with unsparged samples and sparged samples without surfactant. No significant platelet-platelet binding occurred in uncalcified, sparged samples, although 20-30 platelets adhered to bubbles. Without sparging, histamine and adenosine diphosphate provoked platelet-platelet binding with and without surfactants present. Sparging causes platelets to bind to air bubbles and each other. Surfactants added before sparging attenuate platelet-bubble and platelet-platelet binding. Surfactants may have a clinical role in attenuating gas embolism-induced platelet-bubble and platelet-platelet binding.
Efficient biostorage below -150 °C, without sacrificial cryogen
NASA Astrophysics Data System (ADS)
Spoor, Philip S.
2017-12-01
Biostorage is a multi-billion dollar business worldwide and growing rapidly; yet the commercially available options force the user to choose either optimal storage temperature (below -137 °C, the “glass transition temperature” of water) or convenience (no cryogen refill). Passive liquid-nitrogen freezers (storage Dewars with liquid nitrogen pooled at the bottom) provide very cold storage (-190 °C) but the LN2 must be replenished as it boils off. The alternative, so-called “ultra-low” vapor-compression freezers, have no cryogens to replenish and are convenient to use, but only reach storage temperatures above -90 °C. In addition, these tend to be inefficient and costly. Chart Industries is introducing a novel combination of a storage Dewar with a cryocooler (the “Fusion” freezer), that can maintain storage temperatures below -150 °C without the need to replenish any cryogen, while drawing less electricity than any ultra-low on the market. This new product also fits into a relatively narrow “demand window” where on-site cryocooling is not merely more convenient, but also more costeffective than liquid nitrogen delivery.
Timely topics on spent fuel storage
DOE Office of Scientific and Technical Information (OSTI.GOV)
Selin, I.
1994-12-31
The history of spent fuel management in this country has taken several turns, with a final resolution still out of reach. Several repository programs started, stalled ans stopped. The latest effort at Yucca Mountain is progressing but, at best, is years from the early phases of licensing, much less the actual underground disposal of spent fuel. A monitored retrieval storage [MRS] facility was expected to start accepting commercial spent fuel beginning in 1998, but no such facility is clearly on the horizon. All of these recent developments changed the circumstances that we face in spent fuel management. The obvious conclusionmore » is that an increasing number of plants, both operating and permanently shut-down reactors, will have to provide for additional spent fuel storage on-site for a longer period than originally planned, and even after plant decommissioning, prudence requires that provision be made for continual, stand-alone, on-site storage. After pool capacity is reached, most utilities opt for some sort of dry storage. But the dry storage option has triggered an unprecedented amount of local opposition at many sites, further taxing NRC and industry resources.« less
Effect of storage temperature on endogenous GHB levels in urine.
LeBeau, M A; Miller, M L; Levine, B
2001-06-15
Because gamma-hydroxybutyrate (GHB) is an endogenous substance present in the body and is rapidly eliminated after ingestion, toxicologists investigating drug-facilitated sexual assault cases are often asked to differentiate between endogenous and exogenous levels of GHB in urine samples. This study was designed to determine the effects of storage temperature on endogenous GHB levels in urine. Specifically, it was designed to ascertain whether endogenous levels can be elevated to a range considered indicative of GHB ingestion. Urine specimens from two subjects that had not been administered exogenous GHB were collected during a 24h period and individually pooled. The pooled specimens were separated into standard sample cups and divided into three storage groups: room temperature ( approximately 25 degrees C), refrigerated (5 degrees C), and frozen (-10 degrees C). Additionally, some specimens were put through numerous freeze/thaw cycles to mimic situations that may occur if multiple laboratories analyze the same specimen. Periodic analysis of the samples revealed increases in the levels of endogenous GHB over a 6-month period. The greatest increase (up to 404%) was observed in the samples maintained at room temperature. The refrigerated specimens showed increases of 140-208%, while the frozen specimens showed smaller changes (88-116%). The specimens subjected to multiple freeze/thaw cycles mirrored specimens that had been thawed only once. None of the stored urine specimens demonstrated increases in GHB concentrations that would be consistent with exogenous GHB ingestion.
Platelet-derived growth factor inhibits platelet activation in heparinized whole blood.
Selheim, F; Holmsen, H; Vassbotn, F S
1999-08-15
We previously have demonstrated that human platelets have functionally active platelet-derived growth factor alpha-receptors. Studies with gel-filtered platelets showed that an autocrine inhibition pathway is transduced through this tyrosine kinase receptor during platelet activation. The physiological significance of this inhibitory effect of platelet-derived growth factor on gel-filtered platelets activation is, however, not known. In the present study, we investigated whether platelet-derived growth factor inhibits platelet activation under more physiological conditions in heparinized whole blood, which represents a more physiological condition than gel-filtered platelets. Using flow cytometric assays, we demonstrate here that platelet-derived growth factor inhibits thrombin-, thrombin receptor agonist peptide SFLLRN-, and collagen-induced platelet aggregation and shedding of platelet-derived microparticles from the platelet plasma membrane during platelet aggregation in stirred heparinized whole blood. The inhibitory effect of platelet-derived growth factor was dose dependent. However, under nonaggregating conditions (no stirring), we could not demonstrate any significant effect of platelet-derived growth factor on thrombin- and thrombin receptor agonist peptide-induced platelet surface expression of P-selectin. Our results demonstrate that platelet-derived growth factor appears to be a true antithrombotic agent only under aggregating conditions in heparinized whole blood.
Gamonet, Clémentine; Mourey, Guillaume; Aupet, Sophie; Biichle, Sabéha; Petitjean, Régis; Vidal, Chrystelle; Pugin, Aurore; Naegelen, Christian; Tiberghien, Pierre; Morel, Pascal; Angelot-Delettre, Fanny; Seilles, Estelle; Saas, Philippe; Bardiaux, Laurent; Garnache-Ottou, Francine
2017-03-01
The procoagulant and proinflammatory microparticles (MPs) released during storage of packed red blood cells (pRBCs) can potentially modify transfusion benefits. A robust method to quantify MPs in pRBCs is needed to evaluate their impact in clinical trials. The objective was to validate the preanalytic conditions required to prepare pRBC supernatant as well as a method to quantify and evaluate MP variations over 42 days of pRBC storage.A flow cytometry method with size-calibrated beads was developed and fully validated. Quantification of MPs in pRBCs (n = 109) was assessed during short-term (7 days) and long-term (42 days) storage at 4°C, during short-term storage (8 hours) at room temperature, and after 2 years frozen. Repeatability, reproducibility, and linearity of the quantification method were validated, and variations during conservation are presented. There was high variability in RBC (erythrocyte) MP (ERMP) and platelet MP (PMP) levels between RBC units, depending on the filter used for leukocyte reduction. During the 42 days of storage at 4°C, significant increases in ERMPs and PMPs occurred (from 58 to 138 ERMPs/µL from Day 2 to Day 42; p = 0.0002; and from 326 to 771 PMPs/µL from Day 2 to Day 42; p = 0.00026). We use a robust method to confirm that ERMPs and PMPs are present to various degrees in pRBCs and that storage for 42 days significantly increases their generation. This method is robust enough to allow MP quantification in pRBCs and is adapted to evaluate the clinical impact of transfused MPs in prospective clinical trials. © 2017 AABB.
Fibroblast growth factor-2 promotes keratan sulfate proteoglycan expression by keratocytes in vitro
NASA Technical Reports Server (NTRS)
Long, C. J.; Roth, M. R.; Tasheva, E. S.; Funderburgh, M.; Smit, R.; Conrad, G. W.; Funderburgh, J. L.
2000-01-01
Keratocytes of the corneal stroma produce a specialized extracellular matrix responsible for corneal transparency. Corneal keratan sulfate proteoglycans (KSPG) are unique products of keratocytes that are down-regulated in corneal wounds and in vitro. This study used cultures of primary bovine keratocytes to define factors affecting KSPG expression in vitro. KSPG metabolically labeled with [(35)S]sulfate decreased during the initial 2-4 days of culture in quiescent cultures with low serum concentrations (0.1%). Addition of fetal bovine serum, fibroblast growth factor-2 (FGF-2), transforming growth factor beta, or platelet derived growth factor all stimulated cell division, but only FGF-2 stimulated KSPG secretion. Combined with serum, FGF-2 also prevented serum-induced KSPG down-regulation. KSPG secretion was lost during serial subculture with or without FGF-2. Expression of KSPG core proteins (lumican, mimecan, and keratocan) was stimulated by FGF-2, and steady state mRNA pools for these proteins, particularly keratocan, were significantly increased by FGF-2 treatment. KSPG expression therefore is supported by exogenous FGF-2 and eliminated by subculture of the cells in presence of serum. FGF-2 stimulates KSPG core protein expression primarily through an increase in mRNA pools.
Distance Learning and Cloud Computing: "Just Another Buzzword or a Major E-Learning Breakthrough?"
ERIC Educational Resources Information Center
Romiszowski, Alexander J.
2012-01-01
"Cloud computing is a model for the enabling of ubiquitous, convenient, and on-demand network access to a shared pool of configurable computing resources (e.g., networks, servers, storage, applications, and other services) that can be rapidly provisioned and released with minimal management effort or service provider interaction." This…
78 FR 11639 - Dominion Transmission, Inc.; Notice of Request Under Blanket Authorization
Federal Register 2010, 2011, 2012, 2013, 2014
2013-02-19
... under the Natural Gas Act (NGA) as amended, to plug and abandon two storage wells and their associated... plug wells JW-451F and JW-454F and their associated pipelines located near the Murrysville Pool of the.... Dominion further states that the certificated physical parameters, including total natural gas inventory...
Carbon and nitrogen pools in oak-hickory forests of varying productivity
Donald J. Kaczmarek; Karyn S. Rodkey; Robert T. Reber; Phillip E. Pope; Felix, Jr. Ponder
1995-01-01
Carbon (C) and nitrogen (N) storage capacities are critical issues facing forest ecosystem management in the face of potential global climate change. The amount of C sequestered by forest ecosystems can be a significant sink for increasing atmospheric CO2 levels. N availability can interact with other environmental factors such as water...
Long-term effects of wetland harvesting practices on productivity and carbon pools
Scott McKee; Mike Aust; John Seiler; Brian Strahm
2012-01-01
Forested wetlands are valued for social and ecological benefits including filtering sediments, uptake of nutrients, carbon storage, reduction of flood depths, protection of shorelines and streambanks, and provision of terrestrial and aquatic wildlife habitat (Walbridge 1993, Kellison and Young 1997, Brady and Weil 2002). Although the importance of wetland functions are...
Federal Register 2010, 2011, 2012, 2013, 2014
2011-05-10
... the Region I fuel storage racks reflect credit for fuel assembly burnup and soluble boron. Based on... boron concentration of 850 parts per million (ppm) during normal operations, and 1350 ppm during...) racks when considering the presence of soluble boron in the pool water for criticality control and the...
33 CFR 208.32 - Sanford Dam and Lake Meredith, Canadian River, Tex.
Code of Federal Regulations, 2010 CFR
2010-07-01
... Lake Meredith in the interest of flood control as follows: (a) Flood control storage in the reservoir... control pool) initially amounts to 462,100 acre-feet. Whenever the reservoir level is within this... as much as practicable the flood damage below the reservoir. All flood control releases shall be made...
33 CFR 208.32 - Sanford Dam and Lake Meredith, Canadian River, Tex.
Code of Federal Regulations, 2011 CFR
2011-07-01
... Lake Meredith in the interest of flood control as follows: (a) Flood control storage in the reservoir... control pool) initially amounts to 462,100 acre-feet. Whenever the reservoir level is within this... as much as practicable the flood damage below the reservoir. All flood control releases shall be made...
Fire effects on temperate forest soil C and N storage
Lucas E. Nave; Eric D. Vance; Christopher W. Swanston; Peter S. Curtis
2011-01-01
Temperate forest soils store globally significant amounts of carbon (C) and nitrogen (N). Understanding how soil pools of these two elements change in response to disturbance and management is critical to maintaining ecosystem services such as forest productivity, greenhouse gas mitigation, and water resource protection. Fire is one of the principal disturbances acting...
Worldwide organic soil carbon and nitrogen data
DOE Office of Scientific and Technical Information (OSTI.GOV)
Zinke, P.J.; Stangenberger, A.G.; Post, W.M.
The objective of the research presented in this package was to identify data that could be used to estimate the size of the soil organic carbon pool under relatively undisturbed soil conditions. A subset of the data can be used to estimate amounts of soil carbon storage at equilibrium with natural soil-forming factors. The magnitude of soil properties so defined is a resulting nonequilibrium values for carbon storage. Variation in these values is due to differences in local and geographic soil-forming factors. Therefore, information is included on location, soil nitrogen content, climate, and vegetation along with carbon density and variation.
Rupture Forces among Human Blood Platelets at different Degrees of Activation
Nguyen, Thi-Huong; Palankar, Raghavendra; Bui, Van-Chien; Medvedev, Nikolay; Greinacher, Andreas; Delcea, Mihaela
2016-01-01
Little is known about mechanics underlying the interaction among platelets during activation and aggregation. Although the strength of a blood thrombus has likely major biological importance, no previous study has measured directly the adhesion forces of single platelet-platelet interaction at different activation states. Here, we filled this void first, by minimizing surface mediated platelet-activation and second, by generating a strong adhesion force between a single platelet and an AFM cantilever, preventing early platelet detachment. We applied our setup to measure rupture forces between two platelets using different platelet activation states, and blockade of platelet receptors. The rupture force was found to increase proportionally to the degree of platelet activation, but reduced with blockade of specific platelet receptors. Quantification of single platelet-platelet interaction provides major perspectives for testing and improving biocompatibility of new materials; quantifying the effect of drugs on platelet function; and assessing the mechanical characteristics of acquired/inherited platelet defects. PMID:27146004
NASA Astrophysics Data System (ADS)
Scott, D.; Wohl, E.
2017-12-01
The terrestrial organic carbon (OC) pool plays a major role in impacting global climate through the storage and potential release of carbon. In particular, areas of high net primary productivity, such as mountainous regions, and high spatial complexity, such as mountain river floodplains, show potential to act as both strong OC reservoirs and potential OC emitters in a changing climate. We focus on mountain rivers as potential hot swaths of OC storage and, accordingly, as places where land management to retain OC on the landscape may be especially impactful. Mountain river OC storage magnitude and age is a function of the soil and geomorphologic conditions at a reach scale, which are in turn determined by broader characteristics, such as climate, ecology, and tectonics. We present field data on OC storage in soil and wood from three mountain ranges across the western U.S.: the Wind River Range in Wyoming and the Olympic and Central Cascade Ranges in Washington. While the Big Sandy River basin in the Wind River Range exhibits relatively low relief, a semi-arid climate, and a fire-mediated disturbance regime, the Middle Fork Snoqualmie basin in the Cascades and the Sitkum and South Fork Calawah basins in the Olympics exhibit high relief and a humid climate. In contrast to the Olympics, the study basin in the Cascades exhibits strong longitudinal disconnectivity in the form of glaciogenic lakes, whereas the study basins in the Olympics lack large depositional zones that can store sediment for long periods of time. With our expansive dataset of OC storage magnitude and age in downed wood and soil from these three disparate regions, covering a wide range of tectonic, geomorphic, climatic, and ecologic variability, we are able to evaluate both the magnitude and age of the mountain river carbon pool as well as the factors that control that magnitude and age. We present a statistical model that illuminates the dominant controls on the magnitude and age of OC storage in mountain rivers. Using this, we broadly examine mountain river carbon storage dynamics with the goal of allowing land managers to prioritize and focus management efforts to retain OC on the landscape.
Drug-Free Platelets Can Act as Seeds for Aggregate Formation During Antiplatelet Therapy
Hoefer, Thomas; Armstrong, Paul C.; Finsterbusch, Michaela; Chan, Melissa V.; Kirkby, Nicholas S.
2015-01-01
Objective— Reduced antiplatelet drug efficacy occurs in conditions of increased platelet turnover, associated with increased proportions of drug-free, that is, uninhibited, platelets. Here, we detail mechanisms by which drug-free platelets promote platelet aggregation in the face of standard antiplatelet therapy. Approach and Results— To model standard antiplatelet therapy, platelets were treated in vitro with aspirin, the P2Y12 receptor blocker prasugrel active metabolite, or aspirin plus prasugrel active metabolite. Different proportions of uninhibited platelets were then introduced. Light transmission aggregometry analysis demonstrated clear positive associations between proportions of drug-free platelets and percentage platelet aggregation in response to a range of platelet agonists. Using differential platelet labeling coupled with advanced flow cytometry and confocal imaging we found aggregates formed in mixtures of aspirin-inhibited platelets together with drug-free platelets were characterized by intermingled platelet populations. This distribution is in accordance with the ability of drug-free platelets to generate thromboxane A2 and so drive secondary platelet activation. Conversely, aggregates formed in mixtures of prasugrel active metabolite–inhibited or aspirin plus prasugrel active metabolite–inhibited platelets together with drug-free platelets were characterized by distinct cores of drug-free platelets. This distribution is consistent with the ability of drug-free platelets to respond to the secondary activator ADP. Conclusions— These experiments are the first to image the interactions of inhibited and uninhibited platelets in the formation of platelet aggregates. They demonstrate that a general population of platelets can contain subpopulations that respond strikingly differently to overall stimulation of the population and so act as the seed for platelet aggregation. PMID:26272940
High-performance data processing using distributed computing on the SOLIS project
NASA Astrophysics Data System (ADS)
Wampler, Stephen
2002-12-01
The SOLIS solar telescope collects data at a high rate, resulting in 500 GB of raw data each day. The SOLIS Data Handling System (DHS) has been designed to quickly process this data down to 156 GB of reduced data. The DHS design uses pools of distributed reduction processes that are allocated to different observations as needed. A farm of 10 dual-cpu Linux boxes contains the pools of reduction processes. Control is through CORBA and data is stored on a fibre channel storage area network (SAN). Three other Linux boxes are responsible for pulling data from the instruments using SAN-based ringbuffers. Control applications are Java-based while the reduction processes are written in C++. This paper presents the overall design of the SOLIS DHS and provides details on the approach used to control the pooled reduction processes. The various strategies used to manage the high data rates are also covered.
NASA Technical Reports Server (NTRS)
Reed, G. L.; Matsueda, G. R.; Haber, E.
1992-01-01
Platelet clots resist fibrinolysis by plasminogen activators. We hypothesized that platelet factor XIII may enhance the fibrinolytic resistance of platelet-rich clots by catalyzing the crosslinking of alpha 2-antiplasmin (alpha 2AP) to fibrin. Analysis of plasma clot structure by polyacrylamide gel electrophoresis and immunoblotting revealed accelerated alpha 2AP-fibrin crosslinking in platelet-rich compared with platelet-depleted plasma clots. A similar study of clots formed with purified fibrinogen (depleted of factor XIII activity), isolated platelets, and specific factor XIII inhibitors indicated that this accelerated crosslinking was due to the catalytic activity of platelet factor XIII. Moreover, when washed platelets were aggregated by thrombin, there was evidence of platelet factor XIII-mediated crosslinking between platelet alpha 2AP and platelet fibrin(ogen). Specific inhibition (by a monoclonal antibody) of the alpha 2AP associated with washed platelet aggregates accelerated the fibrinolysis of the platelet aggregate. Thus in platelet-rich plasma clots, and in thrombin-induced platelet aggregates, platelet factor XIII actively formed alpha 2AP-fibrin crosslinks, which appeared to enhance the resistance of platelet-rich clots to fibrinolysis.
Genomewide association study of liver abscess in beef cattle.
Keele, J W; Kuehn, L A; McDaneld, T G; Tait, R G; Jones, S A; Keel, B N; Snelling, W M
2016-02-01
Fourteen percent of U.S. cattle slaughtered in 2011 had liver abscesses, resulting in reduced carcass weight, quality, and value. Liver abscesses can result from a common bacterial cause, , which inhabits rumen lesions caused by acidosis and subsequently escapes into the blood stream, is filtered by the liver, and causes abscesses in the liver. Our aim was to identify SNP associated with liver abscesses in beef cattle. We used lung samples as a DNA source because they have low economic value, they have abundant DNA, and we had unrestricted access to sample them. We collected 2,304 lung samples from a beef processing plant: 1,152 from animals with liver abscess and 1,152 from animals without liver abscess. Lung tissue from pairs of animals, 1 with abscesses and another without, were collected from near one another on the viscera table to ensure that pairs of phenotypically extreme animals came from the same lot. Within each phenotype (abscess or no abscess), cattle were pooled by slaughter sequence into 12 pools of 96 cattle for each phenotype for a total of 24 pools. The pools were constructed by equal volume of frozen lung tissue from each animal. The DNA needed to allelotype each pool was then extracted from pooled lung tissue and the BovineHD Bead Array (777,962 SNP) was run on all 24 pools. Total intensity (TI), an indicator of copy number variants, was the sum of intensities from red and green dyes. Pooling allele frequency (PAF) was red dye intensity divided TI. Total intensity and PAF were weighted by the inverse of their respective genomic covariance matrices computed over all SNP across the genome. A false discovery rate ≤ 5% was achieved for 15 SNP for PAF and 20 SNP for TI. Genes within 50 kbp from significant SNP were in diverse pathways including maintenance of pH homeostasis in the gastrointestinal tract, maintain immune defenses in the liver, migration of leukocytes from the blood into infected tissues, transport of glutamine into the kidney in response to acidosis to facilitate production of bicarbonate to increase pH, aggregate platelets to liver injury to facilitate liver repair, and facilitate axon guidance. Evidence from the 35 detected SNP associations combined with evidence of polygenic variation indicate that there is adequate genetic variation in incidence rate of liver abscesses, which could be exploited to select sires for reduced susceptibility to subacute acidosis and associated liver abscess.
Nucleate pool boiling: High gravity to reduced gravity; liquid metals to cryogens
NASA Technical Reports Server (NTRS)
Merte, Herman, Jr.
1988-01-01
Requirements for the proper functioning of equipment and personnel in reduced gravity associated with space platforms and future space station modules introduce unique problems in temperature control; power generation; energy dissipation; the storage, transfer, control and conditioning of fluids; and liquid-vapor separation. The phase change of boiling is significant in all of these. Although both pool and flow boiling would be involved, research results to date include only pool boiling because buoyancy effects are maximized for this case. The effective application of forced convection boiling heat transfer in the microgravity of space will require a well grounded and cogent understanding of the mechanisms involved. Experimental results are presented for pool boiling from a single geometrical configuration, a flat surface, covering a wide range of body forces from a/g = 20 to 1 to a/g = 0 to -1 for a cryogenic liquid, and from a/g = 20 to 1 for water and a liquid metal. Similarities in behavior are noted for these three fluids at the higher gravity levels, and may reasonably be expected to continue at reduced gravity levels.
Liu, Enke; Yan, Changrong; Mei, Xurong; Zhang, Yanqing; Fan, Tinglu
2013-01-01
An understanding of the dynamics of soil organic carbon (SOC) as affected by farming practices is imperative for maintaining soil productivity and mitigating global warming. The objectives of this study were to investigate the effects of long-term fertilization on SOC and SOC fractions for the whole soil profile (0-100 cm) in northwest China. The study was initiated in 1979 in Gansu, China and included six treatments: unfertilized control (CK), nitrogen fertilizer (N), nitrogen and phosphorus (P) fertilizers (NP), straw plus N and P fertilizers (NP+S), farmyard manure (FYM), and farmyard manure plus N and P fertilizers (NP+FYM). Results showed that SOC concentration in the 0-20 cm soil layer increased with time except in the CK and N treatments. Long-term fertilization significantly influenced SOC concentrations and storage to 60 cm depth. Below 60 cm, SOC concentrations and storages were statistically not significant between all treatments. The concentration of SOC at different depths in 0-60 cm soil profile was higher under NP+FYM follow by under NP+S, compared to under CK. The SOC storage in 0-60 cm in NP+FYM, NP+S, FYM and NP treatments were increased by 41.3%, 32.9%, 28.1% and 17.9%, respectively, as compared to the CK treatment. Organic manure plus inorganic fertilizer application also increased labile soil organic carbon pools in 0-60 cm depth. The average concentration of particulate organic carbon (POC), dissolved organic carbon (DOC) and microbial biomass carbon (MBC) in organic manure plus inorganic fertilizer treatments (NP+S and NP+FYM) in 0-60 cm depth were increased by 64.9-91.9%, 42.5-56.9%, and 74.7-99.4%, respectively, over the CK treatment. The POC, MBC and DOC concentrations increased linearly with increasing SOC content. These results indicate that long-term additions of organic manure have the most beneficial effects in building carbon pools among the investigated types of fertilization.
Membrane-lined foundations for liquid thermal storage
NASA Astrophysics Data System (ADS)
Bourne, R. C.
1981-06-01
The membrane lined storage (MLS) container which is a spinoff of vinyl-lined swimming pool and waterbed technologies was developed. The state of development of MLS was evaluated and concepts for MLS structural and heat transfer systems were improved. Preferred structural supports were identified and designed for 1500 gal MLS containers for basement, crawl space, and slab-on-grade foundation types. Techniques are developed to provide space heating via forced air through a finned storage jacket for the two preferred structural enclosure designs. Cost effectiveness of the direct air heating technique is evaluated. Alternate free convection domestic water preheaters and a preferred heat exchanger material is selected. Collector and space heat inlet/outlet designs, design concepts for auxiliary heat input to MLS from resistance electric, combustion, and heat pump sources are developed.
[Influence of raising oxygen content on function of platelet concentrate during preservation].
Zhan, Tong; Xiao, Jian-Yu; Tao, Jing; Miao, Xi-Feng; Liu, Yan-Cun; Tang, Rong-Cai
2006-08-01
To explore the influence of raising oxygen (dissolved oxygen) content on function of platelet concentrate, the platelet concentrate was prepared by a CS-3000 plus blood cell separator. Experiments were divided into 2 groups: test group and control group. After raising oxygen content in platelet plasma under sterile operation, the platelet samples of two groups were preserved in oscillator with horizontal oscillation at 22 +/- 2 degrees C. The platelet count, platelet aggregation rate, lactic acid content and CD62p expression level of platelet were detected on 0, 1, 2, 3, 4, 5 days of platelet preservation. The results showed that the platelet count and platelet aggregation rate decreased with prolongation of preserved time, while the lactic acid content and CD62p expression level of platelet increased gradually. Compared with control group, there were significant differences in aggregation rate of platelet preserved for 2-3 days, and in CD62p expression level of platelet preserved for 1-3 days, while significant difference was found in lactic acid content of platelet preserved for 1-3 days. It is concluded that raising content of oxygen in platelet plasma can provide more oxygen to compensate oxygen supply deficiency for platelet metabolism and improve the efficiency of platelet oxygenic metabolism and the quality of platelet during preservation.
Dismantling the nuclear research reactor Thetis
DOE Office of Scientific and Technical Information (OSTI.GOV)
Michiels, P.
The research reactor Thetis, in service since 1967 and stopped in 2003, is part of the laboratories of the institution of nuclear science of the University of Ghent. The reactor, of the pool-type, was used as a neutron-source for the production of radio-isotopes and for activation analyses. The reactor is situated in a water pool with inner diameter of 3 m. and a depth of 7.5 m. The reactor core is situated 5.3 m under water level. Besides the reactor, the pool contains pneumatic loops, handling tools, graphite blocks for neutron moderation and other experimental equipment. The building houses storagemore » rooms for fissile material and sources, a pneumatic circuit for transportation of samples, primary and secondary cooling circuits, water cleaning resin circuits, a ventilation system and other necessary devices. Because of the experimental character of the reactor, laboratories with glove boxes and other tools were needed and are included in the dismantling program. The building is in 3 levels with a crawl-space. The ground-floor contains the ventilation installation, the purification circuits with tanks, cooling circuits and pneumatic transport system. On the first floor, around the reactor hall, the control-room, visiting area, end-station for pneumatic transport, waste-storage room, fuel storage room and the labs are located. The second floor contains a few laboratories and end stations of the two high speed transfer tubes. The lowest level of the pool is situated under ground level. The reactor has been operated at a power of 150 kW and had a max operating power of 250 kW. Belgoprocess has been selected to decommission the reactor, the labs, storage halls and associated circuits to free release the building for conventional reuse and for the removal of all its internals as legal defined. Besides the dose-rate risk and contamination risk, there is also an asbestos risk of contamination. During construction of the installation, asbestos-containing materials were used, which must be removed in controlled conditions. The ventilation system is considered free from nuclear contamination but it contains asbestos. This paper covers the organization of the dismantling work, the technical execution aspect and conclusions already known (dismantling is ongoing as this is written). (authors)« less
Koranda, M.; Kerschbaum, S.; Wanek, W.; Zechmeister, H.; Richter, A.
2007-01-01
Background and Aims Increased levels of nitrogen (N) deposition lead to enhanced N contents and reduced productivity of many bryophyte species. This study aimed at elucidating the mechanisms by which enhanced N uptake may cause growth reduction of bryophytes, focusing on the effects of N addition on carbon (C) metabolism of bryophytes. Methods Plantlets of Thuidium tamariscinum and Hylocomium splendens were fertilized with NH4NO3 (N load equalling 30 kg ha−1 year−1) for 80 d, including a pulse labelling experiment with 13CO2 to dissect the partitioning of carbon in response to N addition. Key Results Growth of T. tamariscinum was not affected by N addition, while H. splendens showed a trend towards growth reduction. Total N concentration was significantly increased by N addition in H. splendens, a significant increase in amino acid-N was found in T. tamariscinum only. In both bryophyte species, a reduction in concentration of lipids, the greatest C storage pool, as well as markedly enhanced turnover rates of C storage pools in fertilized plants were observed. Conclusions The results suggest that growth reduction of H. splendens under high levels of N deposition may be caused by enhanced synthesis of N-containing organic compounds, most probably of cell wall proteins. Disturbance of cellular C metabolism, as indicated by enhanced C pool turnover, may further contribute to the decline in productivity of H. splendens. PMID:17101638
Ayers, Lisa; Kohler, Malcolm; Harrison, Paul; Sargent, Ian; Dragovic, Rebecca; Schaap, Marianne; Nieuwland, Rienk; Brooks, Susan A; Ferry, Berne
2011-04-01
Circulating cell-derived microparticles (MPs) have been implicated in several disease processes and elevated levels are found in many pathological conditions. The detection and accurate measurement of MPs, although attracting widespread interest, is hampered by a lack of standardisation. The aim of this study was to establish a reliable flow cytometric assay to measure distinct subtypes of MPs in disease and to identify any significant causes of variability in MP quantification. Circulating MPs within plasma were identified by their phenotype (platelet, endothelial, leukocyte and annexin-V positivity (AnnV+). The influence of key variables (i.e. time between venepuncture and centrifugation, washing steps, the number of centrifugation steps, freezing/long-term storage and temperature of thawing) on MP measurement were investigated. Increasing time between venepuncture and centrifugation leads to increased MP levels. Washing samples results in decreased AnnV+MPs (P=0.002) and platelet-derived MPs (PMPs) (P=0.002). Double centrifugation of MPs prior to freezing decreases numbers of AnnV+MPs (P=0.0004) and PMPs (P=0.0004). A single freeze thaw cycle of samples led to an increase in AnnV+MPs (P=0.0020) and PMPs (P=0.0039). Long-term storage of MP samples at -80° resulted in decreased MP levels. This study found that minor protocol changes significantly affected MP levels. This is one of the first studies attempting to standardise a method for obtaining and measuring circulating MPs. Standardisation will be essential for successful development of MP technologies, allowing direct comparison of results between studies and leading to a greater understanding of MPs in disease. Crown Copyright © 2010. Published by Elsevier Ltd. All rights reserved.
Johinke, D; de Graaf, S P; Bathgate, R
2015-08-01
Extending the shelf life of chilled rabbit spermatozoa is vital for the expansion of the farmed rabbit industry. This study evaluated the relationship between sperm concentration and packaging on in vitro quality of chilled rabbit semen over 96 h. Semen was collected from adult bucks (n = 4) and pooled at 37°C following evaluation. Pooled ejaculates were diluted with a Tris-based extender supplemented with 100 μm quercetin to a concentration of 15, 30 or 60 × 10(6) spermatozoa/ml, packaged into plastic tubes or 0.5-ml straws and stored at 15°C. Sperm quality was assessed by computer-assisted sperm Analysis [total motility (tMOT)] and flow cytometry [viability, acrosome integrity, H2 O2 production, plasma membrane disorder, apoptosis and DNA fragmentation index (DFI)] at 0, 48, 72 and 96 h. From 48 h, concentrations of 30 and 60 × 10(6) spermatozoa/ml reported the highest tMOT, irrespective of storage vessel (p < 0.05). Storage in straws reduced oxidative stress and improved plasma membrane stability. The %DFI, mean DFI and SD-DFI were increased in spermatozoa stored in tubes compared with straws (p < 0.05). Although the use of low sperm concentrations in artificial insemination doses would facilitate greater dispersion of genetically superior rabbit bucks, dilution to 15 × 10(6) spermatozoa/ml had a detrimental impact on motility. As such, chilled storage at 30 × 10(6) spermatozoa/ml may provide a suitable balance between motility and H2 O2 production to best maintain overall sperm function and should be evaluated in a large-scale AI trial. © 2015 Blackwell Verlag GmbH.