Comparison Between the Hyperelastic Behavior of Fresh and Frozen Equine Articular Cartilage in Various Joints.

Fresh and frozen cartilage samples of the fetlock, carpus, and stifle were collected from 12 deceased horses. Half were measured immediately following extraction, and half were frozen for seven days and then measured. Seven indentations (various normalized displacements) were implemented with an indention rate of 0.1 mm/s. Solid phase aggregate modulus (Es), hyperelastic material constant (α), and fluid load fraction (F') of equine articular cartilage were assessed using the Ogden hyperelastic model. The properties were statistically compared in various joints (fetlock, carpus, and stifle), and between fresh and frozen samples using various statistical models. There was no statistical difference between the fetlock and carpus in the aggregate modulus (p = 0.5084), while both were significantly different from the stifle (fetlock: p = 0.0017 and carpus: p = 0.0406). For the hyperelastic material constant, no statistical differences between joints were observed (p = 0.3310). For the fluid load fraction, the fetlock and stifle comparison showed a difference (p = 0.0333), while the carpus was not different from the fetlock (p = 0.1563) or stifle (p = 0.3862). Comparison between the fresh and frozen articular cartilage demonstrated no significant difference among the joints in the three material properties: p = 0.9418, p = 0.7031, and p = 0.9313 for the aggregate modulus, the hyperelastic material constant, and the fluid load fraction, respectively.

The spot technique: synthesis and screening of peptide macroarrays on cellulose membranes.

Peptide arrays are a widely used tool for drug development. For peptide-based drug design it is necessary to screen a large number of peptides. However, there are often difficulties with this approach. Most common peptide synthesis techniques are able to simultaneously synthesize only up to a few hundred single peptides. Spot synthesis is a positionally addressable, multiple synthesis technique offering the possibility of synthesizing and screening up to 10,000 peptides or peptide mixtures on cellulose or other membrane surfaces. In this chapter we present the basic procedures and screening methods related to spot synthesis and outline protocols for easy-to-use detection methods on these peptide arrays.

Inactivation of C3a and C5a octapeptides by carboxypeptidase R and carboxypeptidase N.

Pro-carboxypeptidase R (proCPR), also known as thrombin-activatable fibrinolysis inhibitor (TAFI), precursor of carboxypeptidase U and plasma carboxypeptidase B is present in plasma and following activation by thrombin/thrombomodulin and/or plasmin can remove arginine from the carboxyterminal of C3a and C5a. We have shown that this enzyme can remove terminal arginine from the C5a octapeptide much more efficiently than the classical anaphylatoxin inactivator, carboxypeptidase N (CPN). Since we have previously demonstrated that proCPR is significantly upregulated in the inflammatory state, this enzyme would appear to significantly contribute to the inactivation of C5a, the most potent of the complement derived anaphylatoxins.