The sugar cane biofactory - building blocks for the future

The sugarcane plant, with its enormous genetic capacity to accumulate carbon and manufacture and store sucrose, also has the potential to accumulate carbon and metabolically create a wide range of new molecules for industrial and other commercial uses. The extent to which this change can be developed and realised commercially is a function of the technical competence of the industry's R&D capacity, the reality of the commercial drivers which support this global agenda, and the determination of the industry to achieve such goals. The outcomes of existing R&D work already strongly support the technical challenges of this opportunity in sugarcane. The current challenge remains the commercialisation of the technology in a global market in which the current business structures and systems for the manufacture and distribution of existing (competitive) products makes the development of new product lines a higher risk than might otherwise be the case. This is despite all the claims that global markets are expecting and (in some cases) legislating the creation of more sustainable production systems. The options and issues for the development of a sugarcane biofactory system are discussed.

Electrostatically mediated specific adsorption of small molecules in metallo-organic frameworks

We investigate the interaction of ethylene and ethane with a Cu-tricarboxylate complex and show that at low loadings the lighter molecule has a higher binding energy as a result of an increased interaction with the framework Cu and stronger hydrogen bonding with the basic framework oxygens. This leads to selective adsorption of ethylene by a factor of about 2 at low pressure, which is overcome by the stronger van der Waals interaction of ethane at high loadings, explaining recent literature data. The results suggest the possibility of separation of light hydrocarbons at low pressures or in trace amounts.

Mooloolabenes A-E, norsesterterpenes from the Australian sponge Hyattella intestinalis

Five new norsesterterpenes, mooloolabenes A-E (1-5), and the new sesterterpene mooloolaldehyde (6), related to the scalarane family of compounds, were isolated from an acetone extract of the Australian sponge Hyattella intestinalis. Structural elucidation, including relative stereochemical assignment, was based on spectroscopic analysis. All compounds tested showed cytotoxic activity against the P388 cell line.

Why is economics not a complex systems science?

The article argues that economics will have to become a complex systems science before economists can comfortably incorporate institutionalist and evolutionary economics into mainstream theory. The article compares the complex adaptive system of John Foster with that of standard economic theory and illustrates the difference through an examination of familiar production function. The place of neoclassical, Keynesian economics in complex systems is considered. The article concludes that convincing, multiple models have been made possible by the increase in widely available computing power available.

Liberal Democracy and the Inferior 'Other'Other’: Could the peace liberal democracy promises to bring not extend to ‘Others’?

Advocates of liberal democracy argue that its principles and practices contribute directly to peace (at both inter-state and domestic levels). They rely on ideals such as the rule of law, institutional checks and balances on power, an ethos of tolerance, and free market economics to deliver the liberal peace. Liberals, however, overlook three important features embedded in the construction of liberal democracy which can serve to facilitate political violence: 1) the fixed and thus non-negotiable nature of liberal democracy’s core principles, 2) the inferior manner in which it conceives ‘Other’ social orders that do not share its core principles, and 3) the urge to proselytise Others. Together, these constitutive qualities can facilitate moves by leaders of Other groups to argue that liberal democracy threatens ‘their’ preferred identity, and thus its promised peaceful outcomes can be put in doubt.

Overcoming the problem of loss-in-capacity of calcium oxide in CO2 capture

Calcium oxide has been identified to be one of the best candidates for CO2 capture in zero-emission power-generation systems. However, it suffers a well-known problem of loss-in-capacity (i.e., its capacity of CO2 capture decreases after it undergoes cycles of carbonation/decarbonation). This problem is a potential obstacle to the adoption of the new technologies. This paper proposes a method of fabricating a CaO-based adsorbent without the problem of loss-in-capacity. An adsorbent was fabricated using the method and tested on a thermogravimetric analyzer. It was shown that the sorbent attained a utilization efficiency of more than 90% after 9 cycles of carbonation/decarbonation.

The refolding of different alpha-fetoprotein variants

The effect of glycosylation on AFP foldability was investigated by parallel quantitative and qualitative analyses of the refolding of glycosylated and nonglycosylated AFP variants. Both variants were successfully refolded by dialysis from the denatured-reduced state, attaining comparable ``refolded peak'' profiles and refolding yields as determined by reversed-phase HPLC analysis. Both refolded variants also showed comparable spectroscopic fingerprints to each other and to their native counterparts, as determined by circular dichroism spectroscopy. Inclusion body-derived AFP was also readily refolded via dilution under the same redox conditions as dialysis refolding, showing comparable circular dichroism fingerprints as native nonglycosylated AFP. Quantitative analyses of inclusion body-derived AFP showed sensitivity of AFP aggregation to proteinaceous and nonproteinaceous inclusion body contaminants, where refolding yields increased with increasing AFP purity. All of the refolded AFP variants showed positive responses in ELISA that corresponded with the attainment of a bioactive conformation. Contrary to previous reports that the denaturation of cord serum AFP is an irreversible process, these results clearly show the reversibility of AFP denaturation when refolded under a redox-controlled environment, which promotes correct oxidative disulfide shuffling. The successful refolding of inclusion body-derived AFP suggests that fatty acid binding may not be required for the attainment of a rigid AFP tertiary structure, contrary to earlier studies. The overall results from this work demonstrate that foldability of the AFP molecule from its denatured-reduced state is independent of its starting source, the presence or absence of glycosylation and fatty acids, and the refolding method used (dialysis or dilution).

Generic Technique to Generate Large Branched DNA Complexes

The inherent self-recognition properties of DNA have led to its use as a scaffold for various nanotechnology self-assembly applications, with macromolecular complexes, metallic and semiconducting nanoparticles, proteins, inter alia, being assembled onto a designed DNA scaffold. Such structures may typically comprise a number of DNA molecules organized into macromolecules. Many studies have used synthetic methods to produce the constituent DNA molecules, but this typically constrains the molecules to be no longer than around 100 base pairs (30 nm). However, applications that require larger self-assembling DNA complexes, several tens of nanometers or more, need to be generated by other techniques. Here, we present a generic technique to generate large linear, branched, and/or circular DNA macromolecular complexes. The effectiveness of this technique is demonstrated here by the use of Lambda Bacteriophage DNA as a template to generate single- and double-branched DNA structures approximately 120 nm in size.

Dilution versus dialysis - A quantitative study of the oxidative refolding of recombinant human alpha-fetoprotein

Alpha-fetoprotein (AFP) is a commercially important polypeptide with important diagnostic. physiological and immunomodulatory functions. Previous studies into the refolding of this macromolecule are contradictory. and variously suggest that AFP denaturation may be irreversible or that refolding may be achieved by reducing denaturant concentration through dilution but not dialysis. Importantly, these same previous studies do not provide quantitative metrics by which the Success of refolding, and the potential for bioprocess development. can be assessed. Moreover, these same studies do not optimize and control refolding redox potential - an important factor considering that AFP contains 32 cysteines which form 16 disulfide bonds. In this current study, a quantitative comparison of recombinant human AFP (rhAFP) refolding by dilution and dialysis is conducted under optimized redox conditions. rhAFP refolding yields were > 35% (dialysis refolding) and > 75% (dilution refolding) as assessed by RP-HPLC and ELISA, with structural Similarity to the native state confirmed by UV spectroscopy. Dialysis refolding yield was believed to be lower because the gradual reduction in denaturant concentration allowed extended conformational searching. enabling more time for undesirable interaction with other protein molecules and/or the dialysis membrane, leading to a Sub-optimal process outcome. Significant yield sensitivity to redox environment was also observed, emphasizing the importance of physicochemical optimization. This study demonstrates that very high refolding yields can be obtained, for a physiologically relevant protein, with optimized dilution refolding. The study also highlights the quantitative metrics and macromolecular physical spectroscopic 'fingerprints' required to facilitate transition from laboratory to process scale.

Mechanical properties of interfacial films formed by lysozyme self-assembly at the air-water interface

We present the first characterization of the mechanical properties of lysozyme films formed by self-assembly at the air-water interface using the Cambridge interfacial tensiometer (CIT), an apparatus capable of subjecting protein films to a much higher level of extensional strain than traditional dilatational techniques. CIT analysis, which is insensitive to surface pressure, provides a direct measure of the extensional stress-strain behavior of an interfacial film without the need to assume a mechanical model (e.g., viscoelastic), and without requiring difficult-to-test assumptions regarding low-strain material linearity. This testing method has revealed that the bulk solution pH from which assembly of an interfacial lysozyme film occurs influences the mechanical properties of the film more significantly than is suggested by the observed differences in elastic moduli or surface pressure. We have also identified a previously undescribed pH dependency in the effect of solution ionic strength on the mechanical strength of the lysozyme films formed at the air-water interface. Increasing solution ionic strength was found to increase lysozyme film strength when assembly occurred at pH 7, but it caused a decrease in film strength at pH 11, close to the pI of lysozyme. This result is discussed in terms of the significant contribution made to protein film strength by both electrostatic interactions and the hydrophobic effect. Washout experiments to remove protein from the bulk phase have shown that a small percentage of the interfacially adsorbed lysozyme molecules are reversibly adsorbed. Finally, the washout tests have probed the role played by additional adsorption to the fresh interface formed by the application of a large strain to the lysozyme film and have suggested the movement of reversibly bound lysozyme molecules from a subinterfacial layer to the interface.

Assembly of Human Papillomavirus Type-16 Virus-Like Particles: Multifactorial Study of Assembly and Competing Aggregation

Pentameric capsomeres of human papillomavirus capsid protein L1 expressed in Escherichia coli self-assemble into virus-like particles (VLPs) in vitro. A multifactorial experimental design was used to explore a wide range of solution conditions to optimize the assembly process. The degree of assembly was measured using an enzyme-linked immunosorbent assay, and a high-throughput turbidity assay was developed to monitor competing aggregation. The presence of zinc ions in the assembly buffer greatly increased the incidence of aggregation and had to be excluded from the experiment for meaningful analysis. Assembly of VLPs was optimal at a pH of about 6.5, calcium and sodium ions had no measurable effect, and dithiothreitol and glutathione inhibited assembly. Tryptophan fluorescence spectroscopy demonstrated that an increase in urea concentration reduced the rate of VLP formation but had no effect on the final concentration of assembled VLPs. This study demonstrates the use of the hanging-drop vapor-diffusion crystallization method to screen for conditions that promote aggregation and the use of tryptophan fluorescence spectroscopy for real-time monitoring of the assembly process.

Fibre slippage in cyclically perturbed fibrous assemblies

The dynamics of fibre slippage within general non-bonded fibrous assemblies is studied in the situation where the assembly is subjected to general small cyclic loads. Two models are proposed. The first is applicable when the general cyclic loading is complemented by an occasional tugging force on one end of a fibre, which causes it to gradually withdraw from the assembly, such as might occur during the pilling of a textile. The second considers the situation in which the cyclic perturbations act around a constant background load applied to the assembly. The dynamics is reminiscent of self-organized critical behaviour. This model is applied to predict the progressive elongation of a single yarn during weaving.