Andrea L. Woodhead

Preparation and characterization of silica coated iron oxide magnetic nano-particles.

Iron oxide magnetic nano-particles have been prepared by precipitation in an aqueous solution of iron(II) and iron(III) chlorides under basic condition. Surface modifications have been carried out by using tetraethoxysilane (TEOS) and mercaptopropyltrimethoxysilane (MPTMS). The uncoated and coated particles have been characterized with transmission electron microscopy (TEM), energy dispersive X-ray (EDX) spectroscopy, thermal gravimetric analysis (TGA), and infrared (IR) and Raman spectroscopy. The particle sizes as measured from TEM images were found to have mean diameters of 13nm for the uncoated and about 19nm for the coated particles. The measured IR spectra of the uncoated and MPTMS coated particles showed the conversion of magnetite to hematite at high temperature. The results obtained from both IR spectroscopy and TGA revealed that the mercaptopropylsilyl group in the MPTMS coated magnetite decomposed at 600 degrees C and the silica layer of the TEOS coated magnetite was rather stable. Raman spectroscopy has shown the laser heating effect through the conversion of magnetite to maghemite and hematite.

The analysis of merino wool cuticle and cortical cells by Fourier transform Raman spectroscopy

Wool fibers are comprised of proteins known as alpha-keratins and have a complex morphological structure. The major components of this structure, the cuticle and cortical cells, differ in the conformations of their chains as well as their amino acid compositions. High quality Fourier transform Raman spectra of cortical and cuticle cells isolated from fine Merino wool fibers have been obtained. Raman spectroscopy has been shown to be sensitive to the differences in both secondary structure and amino acid composition. The cortical cells were found to be higher in alpha-helical content as compared to the cuticle cells, which had an increased disordered content. Specific information, consistent with amino acid analysis results, regarding cystine, tyrosine, tryptophan, and phenylalnine residues, were obtained for both the cortical and cuticle cells. In addition, the Raman spectra provided information about free thiol groups, amino acids residues with amide group side chains, and residues with protonated carboxyl group side chains. Middle ir transmission spectra of these isolated cells were also obtained. In comparison to the Raman data, the middle ir spectra were found to be not as rich in information.

The effects of stretching on wool fibres as monitored by FT-Raman spectroscopy

Abstract FT-Raman spectroscopy coupled with amide I band deconvolution was used to monitor the conformational changes of the peptide backbone of sulphite pre-treated wool fibres during stretching. The spectral changes observed are consistent with the transition of α-helical to β-pleated sheet structure. These changes, which are most rapid during the early stages of stretching, can be related to protein secondary structure at both the crystalline and molecular levels. Analysis of the amide III region of the spectra reveals that a very small amount of additional disorder is imparted to the peptide backbone as a result of stretching. The consistency in the widths at half-height of the amide I band components implies that stretching does not significantly change the distribution of peptide chain conformations. From the Raman analysis of cells isolated from the stretched fibres, it is evident that conformational changes occur in both the cuticle and cortex. The most evident change, however, is in the cortical cells.

Continuous Production of Flexible Fibers from Transgenically Produced Honeybee Silk Proteins

Flexible and solvent stable fibers are produced after concentrated recombinant honeybee protein solutions are extruded into a methanol bath, dried, drawn in aqueous methanol, then covalently cross-linked using dry heat. Proteins in solution are predominantly coiled coil. Significant levels of non-orientated ß-sheets form during drying or after coagulation in aqueous methanol. Drawing generally aligns the coiled coil component parallel with the fibre axis and ß-sheet component perpendicular to the fiber axis. The fibres are readily handled, stable in the strong protein denaturants, urea and guanidinium, and suitable for a range of applications such as weaving and knitting.

Convergently-evolved structural anomalies in the coiled coil domains of insect silk proteins.

The use of coiled coil proteins as the basis of silk materials is an engineering solution that has evolved convergently in at least five insect lineages-the stinging hymenopterans (ants, bees, hornets), argid sawflies, fleas, lacewings, and praying mantises-and persisted throughout large radiations of these insect families. These coiled coil silk proteins share a characteristic distinct from other coiled coil proteins, in that they are fabricated into solid materials after accumulating as highly concentrated solutions within dedicated glands. Here, we relate the amino acid sequences of these proteins to the secondary and tertiary structural information available from biophysical methods such as X-ray scattering, nuclear magnetic resonance and Raman spectroscopy. We investigate conserved and convergently evolved features within these proteins and compare these to the features of classic coiled coil proteins including tropomyosin and leucine zippers. Our analysis finds that the coiled coil domains of insect silk proteins have several common structural anomalies including a high prevalence of alanine residues in core positions. These atypical features of the coiled coil fibrous proteins - which likely produce deviations from canonical coiled-coil structure - likely exist due to selection pressures related to the process of silk fabrication and the final function of the proteins.

The purple coloration of four late 19th century silk dresses: A spectroscopic investigation

Prior to the 19th century the use of purple dyes for textile coloration was expensive and usually limited to royalty. The discovery of several synthetic purple dyes during the 19th century made the production of purple textiles more affordable and thus more readily available. The identification of the source of the purple coloration is of historical interest. Small yarn samples from four late 19th century silk dresses were analyzed using a combination of thin layer chromatography and surface enhanced Raman spectroscopy, Fourier transform infrared spectroscopy and energy dispersive x-ray spectroscopy. This combination of techniques enabled the analysis of the complex extraction products. While three of the dresses were found to be dyed using methyl violet, the fourth dress was found to be constructed from a warp yarn dyed with methyl violet in the presence of a tannic acid mordant, and a weft yarn dyed with mauve and a tin mordant.

Spectroscopic Assessment of Australian Cotton Waxes

An investigation into the spectroscopic analysis of cotton waxes on Australian cottons was undertaken. The chemical composition of cotton wax is complex and contains a number of lipid classes. Infrared transmission spectroscopy coupled with principal component analysis was found to be capable of discriminating between solvent-extracted cotton waxes with differences in their alkyl functionality. Based on high-performance thin-layer chromatography (HPTLC) results, these differences were associated with an increase in levels of the alkane wax component. On the basis of these results, a photo-acoustic spectroscopic method was developed that could be used to distinguish raw cottons on the basis of these differences. This method was utilized to screen cottons from the Cotton Seed Distributors 2001 seed trial. A preliminary assessment of the scouring and dyeing properties of the various cottons, identified using the photo-acoustic method, was carried out. The results tended to confirm that cottons with increased alkyl functionality, most likely associated with alkane wax, were more difficult to remove and residual wax on the fiber acted as a barrier to dyestuff penetration, thus lowering color yield.

A comparison of convergently evolved insect silks that share β-sheet molecular structure

Raspy crickets produce silk webs that are used to build shelters. These webs have been found to consist of both fiber and film components. Raman spectra obtained from both components were found to be very similar for a given species. The protein structure of the fibers and films produced by both species was predominately β-sheet with lesser amounts of β-turns, unordered and α-helical protein also detected. The orientation of the β-sheet backbone in the fiber was determined to be parallel to the fiber axis. Compared to cocoon and dragline silk the orientation distribution exhibits a significant randomly orientated protein component. Amino acid analysis confirmed the presence of glycine, serine, and alanine in both species, which are known to form antiparallel β-sheet structures. Both species, although at significantly different concentrations, where found to contain proline. This amino acid is uncommon in insect silks, and likely involved in increasing fiber elasticity.

Separation of photo-active nano-crystalline anatase from titanate nanotubes

Photo-active colloidal anatase was prepared from sodium titanate nanotubes by refluxing in 0.3 M HCl. The refluxing was carried out in cycles, replacing the acid each time. After the second reflux cycle a suspension of colloidal anatase was formed above the residual solids. After three reflux cycles all of the nanotubes were converted to a residual mixture of anatase, rutile and brookite. More colloidal anatase could be isolated from this mixture through a series of water rinses. The anatase suspension was found to be made up of particles with an average diameter of 40 nm as well as a very fine 10-15 nm diameter material. This latter dimension is in line with the crystallite size determined from the anatase isolated from the suspension. At pH 1 the anatase suspension was found to be significantly more photo-active in bleaching methyl orange than P25 at exposure times up to 1.5 h. The photo-activity after 1.5 h was found to be 29% higher than the best catalyst prepared by calcining the same titanate nanotube starting material. The increased activities can probably be attributed to the increased surface area, decreased crystallite size and decreased sodium content of the anatase suspension.

Confirmation of Bioinformatics Predictions of the Structural Domains in Honeybee Silk

Honeybee larvae produce a silk made up of proteins in predominantly a coiled coil molecular structure. These proteins can be produced in recombinant systems, making them desirable templates for the design of advanced materials. However, the atomic level structure of these proteins is proving difficult to determine: firstly, because coiled coils are difficult to crystalize; and secondly, fibrous proteins crystalize as fibres rather than as discrete protein units. In this study, we synthesised peptides from the central structural domain, as well as the N- and C-terminal domains, of the honeybee silk. We used circular dichroism spectroscopy, infrared spectroscopy, and molecular dynamics to investigate the folding behaviour of the central domain peptides. We found that they folded as predicted by bioinformatics analysis, giving the protein engineer confidence in bioinformatics predictions to guide the design of new functionality into these protein templates. These results, along with the infrared structural analysis of the N- and C-terminal domain peptides and the comparison of peptide film properties with those of the full-length AmelF3 protein, provided significant insight into the structural elements required for honeybee silk protein to form into stable materials.