Chemical, physical and microbial properties and microbial diversity in manufactured soils produced from co-composting green waste and biosolids.

The effects of adding biosolids to a green waste feedstock (100% green waste, 25% v/v biosolids or 50% biosolids) on the properties of composted products were investigated. Following initial composting, 20% soil or 20% fly ash/river sand mix was added to the composts as would be carried out commercially to produce manufactured soil. Temperatures during composting reached 50 °C, or above, for 23 days when biosolids were included as a composting feedstock but temperatures barely reached 40 °C when green waste alone was composted. Addition of biosolids to the feedstock increased total N, EC, extractable NH(4), NO(3) and P but lowered pH, macroporosity, water holding capacity, microbial biomass C and basal respiration in composts. Additions of soil or ash/sand to the composts greatly increased the available water holding capacity of the materials. Principal component analysis (PCA) of PCR-DGGE 16S rDNA amplicons separated bacterial communities according to addition of soil to the compost. For fungal ITS-RNA amplicons, PCA separated communities based on the addition of biosolids. Bacterial species richness and Shannon's diversity index were greatest for composts where soil had been added but for fungal communities these parameters were greatest in the treatments where 50% biosolids had been included. These results were interpreted in relation to soil having an inoculation effect and biosolids having an acidifying effect thereby favouring a fungal community.

Inverse expression states of the BRN2 and MITF transcription factors in melanoma spheres and tumour xenografts regulate the NOTCH pathway.

The use of adherent monolayer cultures have produced many insights into melanoma cell growth and differentiation, but often novel therapeutics demonstrated to act on these cells are not active in vivo. It is imperative that new methods of growing melanoma cells that reflect growth in vivo are investigated. To this end, a range of human melanoma cell lines passaged as adherent cultures or induced to form melanoma spheres (melanospheres) in stem cell media have been studied to compare cellular characteristics and protein expression. Melanoma spheres and tumours grown from cell lines as mouse xenografts had increased heterogeneity when compared with adherent cells and 3D-spheroids in agar (aggregates). Furthermore, cells within the melanoma spheres and mouse xenografts each displayed a high level of reciprocal BRN2 or MITF expression, which matched more closely the pattern seen in human melanoma tumours in situ, rather than the propensity for co-expression of these important melanocytic transcription factors seen in adherent cells and 3D-spheroids. Notably, when the levels of the BRN2 and MITF proteins were each independently repressed using siRNA treatment of adherent melanoma cells, members of the NOTCH pathway responded by decreasing or increasing expression, respectively. This links BRN2 as an activator, and conversely, MITF as a repressor of the NOTCH pathway in melanoma cells. Loss of the BRN2-MITF axis in antisense-ablated cell lines decreased the melanoma sphere-forming capability, cell adhesion during 3D-spheroid formation and invasion through a collagen matrix. Combined, this evidence suggests that the melanoma sphere-culture system induces subpopulations of cells that may more accurately portray the in vivo disease, than the growth as adherent melanoma cells.