Project description:Primary objectives: The primary objective is to investigate circulating tumor DNA (ctDNA) via deep sequencing for mutation detection and by whole genome sequencing for copy number analyses before start (baseline) with regorafenib and at defined time points during administration of regorafenib for treatment efficacy in colorectal cancer patients in terms of overall survival (OS).
Primary endpoints: circulating tumor DNA (ctDNA) via deep sequencing for mutation detection and by whole genome sequencing for copy number analyses before start (baseline) with regorafenib and at defined time points during administration of regorafenib for treatment efficacy in colorectal cancer patients in terms of overall survival (OS).
Project description:Natural spider silk is one of the world's toughest proteinaceous materials, yet a truly biomimetic spider silk is elusive even after several decades of intense focus. In this study, Next-Generation Sequencing was utilised to produce transcriptomes of the major ampullate gland of two Australian golden orb-weavers, Nephila plumipes and Nephila pilipes, in order to identify highly expressed predicted proteins that may co-factor in the construction of the final polymer. Furthermore, proteomics was performed by liquid chromatography tandem-mass spectroscopy to analyse the natural solid silk fibre of each species to confirm highly expressed predicted proteins within the silk gland are present in the final silk product. We assembled the silk gland transcriptomes of N. plumipes and N. pilipes into 69,812 and 70,123 contigs, respectively. Gene expression analysis revealed that silk gene sequences were among the most highly expressed and we were able to procure silk sequences from both species in excess of 1,300 amino acids. However, some of the genes with the highest expression values were not able to be identified from our proteomic analysis. Proteome analysis of "reeled" silk fibres of N. plumipes and N. pilipes revealed 29 and 18 proteins, respectively, most of which were identified as silk fibre proteins. This study is the first silk gland specific transcriptome and proteome analysis for these species and will assist in the future development of a biomimetic spider silk.
Project description:Due to fascinating mechanical and biological characteristics spider silk is of great interest in many research fields. Among the orb-weavers Nephila edulis is one of the species used as source for natural spider silk in laboratories. Under appropriate conditions, animals can be kept and bred easily. This manuscript gives information about the spiders' natural habitat, behavior, and breeding and compares them with the established methods and conditions within a research laboratory. Keeping conditions and methods of rearing are described in detail. Within a keeping facility with reliable supply of food, cannibalism rate is significantly reduced and spiders mate all year long. Cohabitants of the genus Steatoda are routinely found in laboratory keeping. While these small spiders do not pose a threat to Nephila edulis, cellar spiders (family Pholcidae) have to be extracted as they have been observed hunting for Nephila spiders.
Project description:Spider silks are remarkable materials designed by nature to have extraordinary elasticity. Their elasticity, however, remains poorly understood, as typical stress-strain experiments only allow access to the axial Young's modulus. In this work, micro-Brillouin light spectroscopy (micro-BLS), a noncontact, nondestructive technique, is utilized to probe the direction-dependent phonon propagation in the Nephila pilipes spider silk and hence solve its full elasticity. To the best of our knowledge, this is the first demonstration on the determination of the anisotropic Young's moduli, shear moduli, and Poisson's ratios of a single spider fiber. The axial and lateral Young's moduli are found to be 20.9 ± 0.8 and 9.2 ± 0.3 GPa, respectively, and the anisotropy of the Young's moduli further increases upon stretching. In contrast, the shear moduli and Poisson's ratios exhibit very weak anisotropy and are robust to stretching.