Electro-spinning of pure collagen nano-fibres – just an expensive way to make gelatin?

Scaffolds manufactured from biological materials promise better clinical functionality, providing that characteristic features are preserved. Collagen, a prominent biopolymer, is used extensively for tissue engineering applications, because its signature biological and physico-chemical properties are retained in vitro preparations. We show here for the first time that the very properties that have established collagen as the leading natural biomaterial are lost when it is electro-spun into nano-fibres out of fluoroalcohols such as 1,1,1,3,3,3-hexafluoro-2-propanol or 2,2,2-trifluoroethanol. We further identify the use of fluoroalcohols as the major culprit in the process. The resultant nano-scaffolds lack the unique ultra-structural axial periodicity that confirms quarter-staggered supramolecular assemblies and the capacity to generate second harmonic signals, representing the typical crystalline triple-helical structure. They were also characterised by low denaturation temperatures, similar to those obtained from gelatin preparations ( p > 0.05). Likewise, circular dichroism spectra revealed extensive denaturation of the electro-spun collagen. Using pepsin digestion in combination with quantitative SDS-PAGE, we corroborate great losses of up to 99% of triple-helical collagen. In conclusion, electro-spinning of collagen out of fluoroalcohols effectively denatures this biopolymer, and thus appears to defeat its purpose, namely to create biomimetic scaffolds emulating the collagen structure and function of the extracellular matrix.

Evaluation of novel Streptococcus pyogenes vaccine candidates incorporating multiple conserved sequences from the C-repeat region of the M-protein

A major challenge for Streptococcus pyogenes vaccine development is the identification of epitopes that confer protection from infection by multiple S. pyogenes M-types. Here we have identified and characterised the distribution of common variant sequences from individual repeat units of the C-repeat region (CRR) of M-proteins representing 77 different M-types. Three polyvalent fusion vaccine candidates (SV1, SV2 and SV3) incorporating the most common variants were subsequently expressed and purified, and demonstrated to be alpha-helical by Circular Dichroism (CD), a secondary conformational characteristic of the CRR in the M-protein. Antibodies raised against each of these constructs recognise M-proteins that vary in their CRR, and bind to the surface of multiple S. pyogenes isolates. Antibodies raised against SV1, containing five variant sequences, also kill heterologous S. pyogenes isolates in in vitro bactericidal assays. Further structural characterisation of this construct demonstrated the conformation of SV1 was stable at different pHs, and thermal unfolding of SV1 a reversible process. Our findings demonstrate that linkage of multiple variant sequences into a single recombinant construct overcomes the need to embed the variant sequences in foreign helix promoting flanking sequences for conformational stability, and demonstrates the viability of the polyvalent candidates as global S. pyogenes vaccine candidates.

Analysis of complex molecular systems: The impact of multivariate analysis for resolving the interactions of small molecules with biopolymers - a review

This review is focused on the impact of chemometrics for resolving data sets collected from investigations of the interactions of small molecules with biopolymers. These samples have been analyzed with various instrumental techniques, such as fluorescence, ultraviolet–visible spectroscopy, and voltammetry. The impact of two powerful and demonstrably useful multivariate methods for resolution of complex data—multivariate curve resolution–alternating least squares (MCR–ALS) and parallel factor analysis (PARAFAC)—is highlighted through analysis of applications involving the interactions of small molecules with the biopolymers, serum albumin, and deoxyribonucleic acid. The outcomes illustrated that significant information extracted by the chemometric methods was unattainable by simple, univariate data analysis. In addition, although the techniques used to collect data were confined to ultraviolet–visible spectroscopy, fluorescence spectroscopy, circular dichroism, and voltammetry, data profiles produced by other techniques may also be processed. Topics considered including binding sites and modes, cooperative and competitive small molecule binding, kinetics, and thermodynamics of ligand binding, and the folding and unfolding of biopolymers. Applications of the MCR–ALS and PARAFAC methods reviewed were primarily published between 2008 and 2013.

A curve shortening flow rule for closed embedded plane curves with a prescribed rate of change in enclosed area

Motivated by a problem from fluid mechanics, we consider a generalization of the standard curve shortening flow problem for a closed embedded plane curve such that the area enclosed by the curve is forced to decrease at a prescribed rate. Using formal asymptotic and numerical techniques, we derive possible extinction shapes as the curve contracts to a point, dependent on the rate of decreasing area; we find there is a wider class of extinction shapes than for standard curve shortening, for which initially simple closed curves are always asymptotically circular. We also provide numerical evidence that self-intersection is possible for non-convex initial conditions, distinguishing between pinch-off and coalescence of the curve interior.

Impact response and parametric studies of reinforced concrete circular columns

This paper presents a detailed description of the influence of critical parameters that govern the vulnerability of columns under lateral impact loads. Numerical simulations are conducted by using the Finite Element program LS-DYNA, incorporating steel reinforcement, material models and strain rate effects. A simplified method based on impact pulse generated from full scale impact tests is used for impact reconstruction and effects of the various pulse loading parameters are investigated under low to medium velocity impacts. A constitutive material model which can simulate failures under tri-axial state of stresses is used for concrete. Confinement effects are also introduced to the numerical simulation and columns of Grade 30 to 50 concrete under pure axial loading are analysed in detail. This research confirmed that the vulnerability of the axially loaded columns can be mitigated by reducing the slenderness ratio and concrete grade, and by choosing the design option with a minimal amount of longitudinal steel. Additionally, it is evident that approximately a 50% increase in impact capacity can be gained for columns in medium rise buildings by enhancing the confinement effects alone. Results also indicated that the ductility as well as the mode of failure under impact can be changed with the volumetric ratio of lateral steel. Moreover, to increase the impact capacity of the vulnerable columns, a higher confining stress is required. The general provisions of current design codes do not sufficiently cover this aspect and hence this research will provide additional guidelines to overcome the inadequacies of code provisions.

Twisted Edwards curves revisited

This paper introduces fast algorithms for performing group operations on twisted Edwards curves, pushing the recent speed limits of Elliptic Curve Cryptography (ECC) forward in a wide range of applications. Notably, the new addition algorithm uses for suitably selected curve constants. In comparison, the fastest point addition algorithms for (twisted) Edwards curves stated in the literature use . It is also shown that the new addition algorithm can be implemented with four processors dropping the effective cost to . This implies an effective speed increase by the full factor of 4 over the sequential case. Our results allow faster implementation of elliptic curve scalar multiplication. In addition, the new point addition algorithm can be used to provide a natural protection from side channel attacks based on simple power analysis (SPA).

Jacobi quartic curves revisited

This paper provides new results about efficient arithmetic on Jacobi quartic form elliptic curves, y 2 = d x 4 + 2 a x 2 + 1. With recent bandwidth-efficient proposals, the arithmetic on Jacobi quartic curves became solidly faster than that of Weierstrass curves. These proposals use up to 7 coordinates to represent a single point. However, fast scalar multiplication algorithms based on windowing techniques, precompute and store several points which require more space than what it takes with 3 coordinates. Also note that some of these proposals require d = 1 for full speed. Unfortunately, elliptic curves having 2-times-a-prime number of points, cannot be written in Jacobi quartic form if d = 1. Even worse the contemporary formulae may fail to output correct coordinates for some inputs. This paper provides improved speeds using fewer coordinates without causing the above mentioned problems. For instance, our proposed point doubling algorithm takes only 2 multiplications, 5 squarings, and no multiplication with curve constants when d is arbitrary and a = ±1/2.

Faster group operations on elliptic curves

This paper improves implementation techniques of Elliptic Curve Cryptography. We introduce new formulae and algorithms for the group law on Jacobi quartic, Jacobi intersection, Edwards, and Hessian curves. The proposed formulae and algorithms can save time in suitable point representations. To support our claims, a cost comparison is made with classic scalar multiplication algorithms using previous and current operation counts. Most notably, the best speeds are obtained from Jacobi quartic curves which provide the fastest timings for most scalar multiplication strategies benefiting from the proposed 12M + 5S + 1D point doubling and 7M + 3S + 1D point addition algorithms. Furthermore, the new addition algorithm provides an efficient way to protect against side channel attacks which are based on simple power analysis (SPA). Keywords: Efficient elliptic curve arithmetic,unified addition, side channel attack.

Faster pairings on special Weierstrass curves

This paper presents efficient formulas for computing cryptographic pairings on the curve y 2 = c x 3 + 1 over fields of large characteristic. We provide examples of pairing-friendly elliptic curves of this form which are of interest for efficient pairing implementations.