Measurement of activated factor IX in factor IX concentrates: correlation with in vivo thrombogenicity.

Current in vitro tests for thrombogenicity of FIX concentrates used for prothrombin complex concentrates (PCCs), are of little value when applied to high purity FIX (HP FIXs). In the present study, we have developed a chromogenic assay for activated FIX (FIXa) and evaluated its ability to predict in vivo thrombogenic potential of HP FIXs in a modified Wessler stasis model. Among the HP FIXs, only 1 out of 7 products had no detectable FIXa; this product also showed no in vivo thrombogenicity.

Tyrosine kinase JAK1 is associated with the granulocyte-colony-stimulating factor receptor and both become tyrosine-phosphorylated after receptor activation.

Granulocyte-colony-stimulating factor (G-CSF) stimulates the proliferation and differentiation of cells of the neutrophil lineage by interaction with a specific receptor. Early signal transduction events following G-CSF receptor activation were studied. We detected tyrosine phosphorylation of both the G-CSF receptor and the protein tyrosine kinase JAK1 following G-CSF binding to the human G-CSF receptor.

Stabilization of von Willebrand factor in banked blood by leucocyte depletion.

The von Willebrand factor (vWf) activity, as measured by the ristocetin co-factor (vWf:RCo) and collagen binding (vWf:CBA) assays, declined progressively in standard blood units stored at 4 degrees C after a 2-day storage period. This loss of activity was accompanied by a loss and degradation of high molecular weight (HMW) vWf multimers. In studies using a paired design, filtration of blood with a high efficiency leucocyte-removal filter, prior to storage at 4 degrees C, led to significantly improved maintenance of vWf:RCo and vWf:CBA compared with unfiltered units (P < 0.

Modulation of fibrinogen content in cryoprecipitate by temperature manipulation during plasma processing.

In routine blood bank production of single-donation cryoprecipitate, the introduction of a 16-hour hold at 4 degrees C, with the frozen plasma units packed into polystyrene containers, resulted in plasma prethaw temperatures of -4 degrees C to -8 degrees C. This in turn resulted in cryoprecipitate fibrinogen levels that were 214 percent of those obtained when units were thawed immediately after removal from -30 degrees C storage. In scale-model production of factor VIII concentrate, plasma warmed from -30 to -10 to -15 degrees C over 18 hours before pooling and thawing yielded cryoprecipitate fibrinogen levels that were 66 percent of those found in plasma warmed to -2 to -5 degrees C over the same period.

The effects of oxytocin on the pulmonary and systemic circulation in pregnant ewes.

The haemodynamic effects of oxytocin on the pulmonary and systemic circulation were studied in six awake, pregnant (greater than 140 days gestation) ewes. Bolus doses of oxytocin 0.2 units/kg and then 0.

von Willebrand factor characterization of a severe dry-heat treated factor VIII concentrate, AHF (high purity).

Eight batches of a severe dry-heat treated (80 degrees C for 72 hours) Factor VIII concentrate manufactured by the Commonwealth Serum Laboratories (CSL Ltd.) were analysed for the following von Willebrand factor-related (vWf) activities: ristocetin cofactor activity (vWf:RCof), collagen binding activity (CBA), vWf antigen levels (vWf:Ag), vWf multimeric analysis and 2-stage FVIII clotting activity (VIII:C). The average potency per vial of vWf:Ag was 440 +/- 80 units, vWf:RCof 500 +/- 60 units, CBA 350 +/- 50 units and VIII:C 242 +/- 36 International Units.

Effect of fetal haemoglobin on the accuracy of pulse oximetry in preterm infants.

Pulse oximeters are programmed with a calibration curve derived from studies done in adults. Whether fetal haemoglobin levels affect their reliability is unclear. This study reports the accuracy of pulse oximetry in 22 preterm infants (mean 31 weeks, range 25-36 weeks gestation) between 1 h and 73 days of age.

The binding and regulation of protein S by neutrophils.

The degradation of the coagulation cofactors Va and VIIIa by activated protein C (APC), is dependent on calcium, a suitable surface and protein S. The latter is a vitamin K-dependent protein which functions as a cofactor in the reaction. In this study we investigated the role of neutrophils in regulating the activity of protein S.

Effects of plasma collection systems and processing parameters on the quality of factor IX concentrate.

Using pilot-scale production of our present factor IX (II and X) concentrate, we have studied the effects of starting plasma source and processing parameter on two in-vitro indicators of product quality - yield and thrombogenic potential. Plasma source did not affect factor IX yield but had a marked effect on thrombogenic potential. Factor IX concentrates produced from plasma derived through centrifugation-based technology showed significantly higher thrombogenic potential than products derived from plasma derived through a filtration-based system.

Comparison of two dimensional immunoelectrophoresis and multimer analysis in the study of von Willebrand factor.

To assess the validity of the multimeric analysis of von Willebrand factor (vWf), this technique was compared with two dimensional immunoelectrophoresis using samples of purified vWf obtained by gel filtration and plasma samples from a patient with severe von Willebrands disease who was receiving prophylaxis with cryoprecipitate. It is concluded that except for specialised clinical and research purposes, two dimensional immunoelectrophoresis provides a clear picture of the multimeric composition of the vWf molecule, and that this is sufficient for routine research and clinical use, without having to confirm the data by multimeric analysis.

The regulation of human factor V by a neutrophil protease.

Neutrophils activated with serum opsonized zymosan, soluble heat-aggregated IgG, and ionophore A23187 in the presence of calcium release a material capable of initially activating factor V. Subsequent inactivation of factor V was only observed with neutrophil releasate derived from IgG and ionophore. In this study we examine the nature of this neutrophil activity and investigate its role in the regulation of factor V/Va.