Serine protease inhibitors (serpins)

Inhibition of serine proteases by serpins (serpin: serine protease inhibitor) is a key mechanism for the control of proteolysis in thrombosis, shock, and inflammation. The various members of the serpin gene superfamily (α(1)-antitrypsin, ovalbumin, C1-inhibitor, antithrombin III, α(2)-antiplasmin, type-1 plasminogen-activator inhibitor, and so forth) have many characteristics in common. In this article, we review the biochemistry and cell biology of serpins, and we discuss their clinical importance and therapeutic potential.     

Attention in musicians is more bilateral than in non-musicians

Attention in neurologically intact adults normally errs towards the left side of space, as documented in studies involving tasks of visual attention (i.e., line bisection). The aim of this study was to further investigate lateralisation of attention in musicians and non-musicians. Reaction times and accuracy were recorded to stimuli presented to the left and right of a vertical line in 20 right-handed musicians and 20 matched non-musician controls. While both groups performed more accurately to left-sided stimuli, performance by the musician group was significantly more accurate than the non-musician group for the right-sided stimuli. Musicians also had faster reaction times overall. Consistent with previous research, the results indicate a more balanced attentional capacity in musicians, as well as enhanced visuomotor ability, and are interpreted with reference to extended musical training.     

Inhibition of plasma kallikrein by C1-inhibitor: role of endothelial cells and the amino-terminal domain of C1-inhibitor

Activation of plasma prekallikein and generation of bradykinin are responsible for the angioedema attacks observed with C1-inhibitor deficiency. Heterozygous individuals with <50% levels of active C1-inhibitor are susceptible to angioedema attacks indicating a critical need for C1-inhibitor to be present at maximum levels to prevent unwanted prekallikrein activation. Studies with purified proteins do not adequately explain this observation. Therefore to investigate why reduction of C1-inhibitor to levels seen in angioedema patients results in excessive kallikrein generation we examined the effect of endothelial cells on the inhibition of kallikrein by C1-inhibitor. Surprisingly, it was found that a C1-inhibitor concentration of greater than 1 microM was needed to inhibit 3 nM kallikrein. We propose that this apparent protection from inhibition was mediated by kallikrein binding to the cells via the heavy chain in a high molecular weight kininogen and zinc independent manner. Protection of kallikrein from inhibition was not observed when C1-inhibitor truncated in the amino-terminal domain by the StcE metalloproteinase was used, which suggests a novel function for this unique domain. The requirement for high concentrations of C1-inhibitor to fully inhibit kallikrein is consistent with the fact that reduced levels of C1-inhibitor result in the kallikrein activation seen in angioedema.     

The inhibition of TNK-t-PA by C1-inhibitor.

TNK-t-PA is a recombinant mutant of tissue plasminogen activator that has a longer half-life and higher selectivity for fibrin than normal tissue plasminogen activator (t-PA). In addition, it is reported to be serpin resistant because of reduced inhibition by plasminogen activator inhibitor-1. In this study, we have investigated the inhibition of TNK-t-PA by the serpin C1-inhibitor. TNK-t-PA is inhibited with a second-order rate constant of 7.5 per mol/l per s compared with 4.5 per mol/l per s for t-PA. In both cases, the stoichiometry was close to 20, indicating that C1-inhibitor was predominantly a substrate for both forms of t-PA. The formation of cleaved C1-inhibitor was seen on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the t-PA-C1-inhibitor (or TNK-t-PA-C1-inhibitor) complex seen on SDS-PAGE and by zymography. Although the rates of inhibition are very slow in vitro, the fact that in vivo formation of the t-PA-C1-inhibitor complex after infusion of t-PA has been well documented suggests that TNK-t-PA will also be inhibited by C1-inhibitor in vivo and, perhaps more importantly, could cause a significant reduction in C1-inhibitor concentration.     

Low-affinity heparin stimulates the inactivation of plasminogen activator inhibitor-1 by thrombin

The influence of heparin on the reaction between thrombin and plasminogen activator inhibitor-1 (PAI-1) has been examined. With a 50- fold excess of PAI-1, the rate constant for the inhibition of thrombin was 458 mol/L-1s-1, which increased to 5,000 mol/L-1s-1 in the presence of 25 micrograms/mL unfractionated heparin or heparin with low affinity for antithrombin. The effect of low affinity heparin was then examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, using close to equimolar concentrations of reactants. Thrombin and PAI-1 formed a stable stoichiometric complex in the absence of heparin, which did not dissociate after the addition of 25 micrograms/mL low-affinity heparin. In contrast, when low-affinity heparin was added at the beginning of the reaction, there was an initial increase in PAI-1- thrombin complex formation, but this was rapidly followed by substantial proteolytic cleavage of unreacted PAI-1 and of the thrombin- PAI-1 complex. The idea that the relative concentrations of thrombin and PAI-1, and the presence of low affinity heparin, could influence the products of the reaction was examined in detail. Quantitative zymographic analysis of tissue plasminogen activator and PAI-1 activities and chromogenic substrate assay of thrombin activity showed that low-affinity heparin stimulated the inactivation of PAI-1 by an equimolar amount of thrombin, but caused only a minimal stimulation of thrombin inhibition. It is concluded that low-affinity heparin stimulates thrombin inhibition when PAI-1 is in excess, but, unexpectedly, that low-affinity heparin enhances PAI-1 inactivation when thrombin is equimolar to PAI-1.     

The reaction between plasmin and C1-inhibitor results in plasmin inhibition by the serpin mechanism.

C1-inhibitor is an important inhibitor of plasma kallikrein and C1, but also has inhibitory activity against numerous other plasma proteinases such as plasmin. The relevance of plasmin inhibition by the C1-inhibitor has been debated, with some evidence showing that plasmin causes significant proteolysis of C1-inhibitor. In the present study, we show that C1-inhibitor in its native state will inhibit plasmin without being significantly degraded, in a manner typical of all serpin reactions. However, if C1-inhibitor is in a denatured polymeric state (as can easily occur during storage, or as produced by heating of the native protein), it will be extensively degraded by plasmin. In addition, we show that hydrophobic interaction chromatography is an effective method to remove trace contaminants of inactive C1-inhibitor polymers.     

Constructive Functional Diversity: A new paradigm beyond disability and impairment

Aims of the paper. This article presents a more dynamic and constructive paradigm than the current dominant ones (for example medical or social models), to describe and change the impact of impairment and disability. The reflections contained are inspired by personal and professional frustration with the existing polarized ideology of human function, which fails to adequately describe the diversity of physiological and psychosocial function amongst people. It aims to provoke and inspire dialogue about our current paradigm of human function in relation to value and capacity.

Key findings and implications. Within this paper: I critique society's biases regarding of functional deficit relative to the subconscious fear of losing function; I question the polarity of the negatively framed language of impairment and disability; I offer constructive, creative ‘solutions’ to describe the experience of atypical function. In so doing, an entirely new language of diverse human function and a concept of Constructive Functional Diversity (CFD) is proposed, which includes a complex yet logical array of modes and outcomes of function.

Conclusions and recommendations. Finally I suggest the benefits of a more dynamic paradigm of functional change in enhancing rehabilitative outcomes, including client-directed practice.