Paired comparison of methylene blue- and amotosalen-treated plasma and cryoprecipitate.

BACKGROUND AND OBJECTIVES: Cryoprecipitate is used in the treatment of patients with acquired hypofibrinogenaemia. Studies have not directly compared cryoprecipitate produced following pathogen inactivation (PI) of fresh-frozen plasma (FFP) using different systems. The effects of methylene blue (MB) and amotosalen (AS) PI systems on the quality of FFP and cryoprecipitate were investigated in a paired study. MATERIALS AND METHODS: Seven group A and 7 group O pools of plasma were prepared and split into individual units and rapidly frozen to produce FFP. Units of FFP were thawed and either PI treated with MB or amotosalen, or left untreated (control). Samples of FFP along with the corresponding cryoprecipitate were tested for a range of coagulation factors, thrombin generation (TGT) and rotational thromboelastometry (ROTEM). RESULTS: AS-FFP showed a smaller decrease following treatment for most coagulation factors analysed than MB-FFP, except fibrinogen (antigen) and factor VII, partly due to lower volume losses. There was no significant difference between treatment methods for fibrinogen content of cryoprecipitate with treated units meeting current UK specification, or TGT and ROTEM parameters studied. CONCLUSIONS: MB-cryo contained a significantly higher content of FVIII and lower content of FXIII when compared to AS-cryo, with no difference in fibrinogen activity.

Thrombin generation and coagulation factor content of thawed plasma and platelet concentrates.

BACKGROUND: We assessed the haemostatic capacity of thawed plasma produced after ambient storage of whole blood for 24 h (RTFP24), and the supernatant of buffy-coat derived platelet concentrates (PC). METHODS: Platelet concentrates (n = 20) were tested on days 1, 5 and 7 of storage at 22°C and RTFP24 (n = 10) immediately following thawing and after 4 and 6 days storage at 4°C. Coagulation factor activity, thrombin generation ± an activator of protein C (PROTAC) and rotational thromboelastometry (ROTEM) were assessed. RESULTS: In plasma and buffy-coat derived PC, there was a < 10% loss of factors II, IX and FX, but much higher loss of factors FV, FVII and FVIII. In plasma, the total or peak amount of thrombin generated was unaffected by storage for 6 days, with or without Protac, but there was an increase in lag time and decreased rate of clot formation by ROTEM. In PC, but not plasma, there was a 16% increase in FXII activity and increase in resistance to activated protein C, co-incidental to 30% loss of free protein S. CONCLUSIONS: These data suggest thrombin generation is relatively unaltered when RTFP24 is thawed and stored for 6 days, and that the supernatant of PC has significant haemostatic capacity.

The "E" word: epilepsy and perceptions of unfair treatment from the 2010 Australian Epilepsy Longitudinal Survey.

AIM: The aim of the current study was to examine self-report data on perceptions of unfair treatment due to epilepsy. METHOD: We analyzed data from the 2010 Australian Epilepsy Longitudinal Survey, distributed to 621 registrants on the Australian Epilepsy Research Register. A total of 343 responses were received (55% response rate), providing insight into experiences of life with epilepsy in Australia. Responses relating to perceptions of unfair treatment in areas of employment, education and community participation as a result of epilepsy are reported in this article. RESULTS: Forty-eight percent of respondents reported perceptions of unfair treatment as a result of their epilepsy, with most providing details of their experiences. Discrimination in the workplace remains of key concern, with 47% citing examples of unfair treatment in this setting. CONCLUSIONS: In spite of Australian anti-discrimination laws, findings indicate that full-time employment rates for people with epilepsy are lower than previously reported, with further mechanisms for support required to improve education and reduce experiences of stigma.

Coagulation factor content of plasma produced from whole blood stored for 24 hours at ambient temperature: results from an international multicenter BEST Collaborative study.

BACKGROUND: There is increasing international interest in producing components from blood that has been stored at room temperature for 24 hours. The lack of comprehensive data on the quality of plasma produced from blood stored in this way led to this international study. STUDY DESIGN AND METHODS: A total of 128 units of whole blood were pooled in groups of four and split to produce 32 sets of four identical blood units that were processed either within 8 hours of blood collection or after 24-hour storage at 18 to 25°C. RESULTS: Storage of whole blood for 24 hours resulted in a 23% decrease in the activity of Factor (F)VIII, but not significant loss of activity of coagulation factors FV, FVII, FXI, FXII, fibrinogen, antithrombin, or von Willebrand factor. There was a small, but significant decrease in levels of FII, FIX, and FX (all <5%) as well as protein C (6%) and free protein S activity (14%). The ability of plasma to generate thrombin after 24-hour storage as whole blood was unaltered, as assessed by real-time thrombin generation tests as was the rate and strength of clot formation by rotational thombelastometry. Levels of all coagulation factors measured were above 0.50 U/mL in plasma produced from whole blood stored for 24 hours. CONCLUSION: These data show that there is minimal effect of storing whole blood at ambient temperature for 24 hours on the coagulation activity of plasma and that this is an acceptable alternative to producing plasma on the day of blood collection.

Platelet apoptosis and activation in platelet concentrates stored for up to 12 days in plasma or additive solution.

BACKGROUND: Several studies suggest that apoptosis of platelets occurs during storage of platelet concentrates (PC). We sought to determine whether storage of PC in additive solution alters levels of apoptosis during storage beyond the current shelf life (5-7 days). STUDY DESIGN AND METHODS: Pooled buffy coat PC (n = 7) were prepared in either 100% plasma or 70% Composol and stored at 22 °C for 12 days. A third arm of the study stored PC in 100% plasma at 37 °C, which is thought to induce apoptosis. PC were tested for mitochrondrial membrane potential, annexin V binding, microparticles, caspase-3/7 activity and decoy cell death receptor 2, as well as standard platelet quality tests. RESULTS: Composol units remained ≥pH 6·88, with 36% lower lactate and higher pH vs plasma by day 12 (P < 0·001). Platelet function was better maintained, and activation and apoptotic markers tended to be lower in Composol units towards the end of storage. However, levels of all apoptosis markers assessed were not significantly different in units stored in Composol. Storage at 37 °C saw stronger correlation of apoptotic markers with standard quality tests compared to 22 °C, but loss of correlation of caspase-3/7 activity with other apoptosis markers. CONCLUSION: We conclude that storage of platelets in 70% Composol vs 100% plasma does not increase the rate of platelet apoptosis. Our data agree with other studies suggesting that platelet apoptosis is sequential to high levels of activation, but share a significant degree of overlap.

A simple spectrophotometric method for the quantification of residual haemoglobin in platelet concentrates.

BACKGROUND AND OBJECTIVES: High levels of residual haemoglobin (Hb 0.1 g/l) are known to decrease the efficiency of pathogen-inactivation systems. We evaluated three separate methods to quantify Hb in platelet concentrates (PC). MATERIALS AND METHODS: Nine PC prepared in platelet additive solution (PASIII) (median platelet yield of 283 x 10(9)/unit, range 46-353) were spiked to known Hb concentrations with whole blood and the samples were measured by using each of three methods: the 3,3',5,5'-tetramethylbenzidine (TMB) oxidation method (Sigma Diagnostics, 527-A); the Harboe spectrophotometric method; and the HemoCue plasma low-Hb photometer (PLHP). RESULTS: The TMB and Harboe methods showed linear results compared to expected Hb (r2 > or = 0.981, P < 0.001) over the range tested (0.09-0.28 g/l) when the samples were haemolysed. The TMB method underestimated by an average of 6%, at and around 0.1 g/l Hb, compared to a 4% overestimation by the Harboe method and a threefold overestimation by the PLHP. The Harboe intra-assay coefficient of variation was < or = 1.85% across all concentrations, which contrasted with 30% at and around 0.1 g/l for the TMB method. CONCLUSIONS: The Harboe spectrophotometric method is convenient, safe, accurate and reproducible, and outperforms the TMB and PLHP methods for quantification of residual Hb in PC.

Anti-arthritic activity in rats of some phosphinegold(i) thionucleobases and related thiolates.

A number of phosphinegold(I) thiolates where, generally, the thiolate is derived from a thionucleobase, have been screened for anti-arthritic activity in Dark Agouti rats, a gold sensitive model for arthritis. Potency and toxicity data showed that, generally, the Ph(3)P derivatives and species based on thiopurines were the most effective and that with other complexes enhanced activity was accompanied by greater toxicity.

In vitro antitumour activity of some triorganophosphinegold(i) thionucleobases.

A series of phosphinegold(I) thionucleobase analogues, [R(3)PAu(SR(x))] (R = Et, Ph or chexyl; HSR(1) = 2-mercaptobenzoic acid, HSR(2) = 2-thiouracil, HSR(3) = 6-mercaptopurine and HSR(4) = 6-thioguanine) have been examined for their in vitro cytotoxicity in L1210 murine leukemia cells in culture. The range of ID(50) values (continuous 48 h exposure) for the complexes is 0.041 - 0.131 muM. The complexes with SR(3) and SR(3) are generally the most active; however, there is no clear trend associated with the phosphine ligands.

Increases in cell elongation, plastid compartment size and phytoene synthase activity underlie the phenotype of the high pigment-1 mutant of tomato.

A characteristic trait of the high pigment-1 ( hp-1) mutant phenotype of tomato ( Lycopersicon esculentum Mill.) is increased pigmentation resulting in darker green leaves and a deeper red fruit. In order to determine the basis for changes in pigmentation in this mutant, cellular and plastid development was analysed during leaf and fruit development, as well as the expression of carotenogenic genes and phytoene synthase enzyme activity. The hp-1 mutation dramatically increases the periclinal elongation of leaf palisade mesophyll cells, which results in increased leaf thickness. In addition, in both palisade and spongy mesophyll cells, the total plan area of chloroplasts per cell is increased compared to the wild type. These two perturbations in leaf development are the primary cause of the darker green hp-1 leaf. In the hp-1 tomato fruit, the total chromoplast area per cell in the pericarp cells of the ripe fruit is also increased. In addition, although expression of phytoene synthase and desaturase is not changed in hp-1 compared to the wild type, the activity of phytoene synthase in ripe fruit is 1.9-fold higher, indicating translational or post-translational control of carotenoid gene expression. The increased plastid compartment size in leaf and fruit cells of hp-1 is novel and provides evidence that the normally tightly controlled relationship between cell expansion and the replication and expansion of plastids can be perturbed and thus could be targeted by genetic manipulation.