Project description:Filamentous algae (FA) have potential advantages over microalgae for wastewater treatment. However, their implementation at large-scale is hindered by an inability to predict performance. This study compared the cellular responses (photosynthesis and respiration) and composition (pigments and photosystem proteins) of FA Oedogonium acclimatised to average summer and winter conditions (Melbourne, Australia). After 7 days of acclimation the Chl a content of summer acclimated (SA) algae was about half that of the winter acclimated (WA) algae, which can be related to a strategy to reduce photodamage under high light intensities. No statistically significant changes were observed in any identified proteins associated with photosystem PSII and the reaction centre of PSI. Transmission electron microscopy images revealed more prominent lipid bodies within the SA filaments than in WA filaments, but no discernible difference in the abundance of starch granules. Photosynthetic irradiance curves were compared for the SA and WA algae. Consistent with the differences in chlorophyll, the specific gross photosynthetic rate (µP, gross) was generally higher for the WA algae. The relative difference increased from around 2-fold at 15°C to 3-fold at 25°C, and then decreased to less than 1.5-fold at 30 °C and 35 °C. At all the tested temperatures, saturation irradiance levels were in the range of 75 – 500 µmol/m2·s. Photoinhibition was observed at 30 °C (above ~300 µmol/m2·s) and was more severe at 35 °C (above ~500 µmol/m2·s), with WA algae showing greater inhibition. In contrast, the respiration response was similar for the SA and WA algae. The study emphasises the significance of accounting for seasonal variations and their effects on biomass productivity and utilisation. The data obtained will enable the incorporation of acclimation and its effect on biochemistry and photosynthetic response into predictive models of FA performance in outdoor cultures.
Project description:The inflammatory functions of the cytokine tumor necrosis factor (TNF) rely on its ability to induce cytokine production and to induce cell death. Caspase dependent and independent pathways – apoptosis and necroptosis – respectively, regulate immunogenicity by the release of distinct sets of cellular proteins. To obtain an unbiased, systems-level understanding of this important process, we here applied mass spectrometry-based proteomics to dissect protein release during apoptosis and necroptosis. We report hundreds of proteins released from human myeloid cells in time-course experiments. Both cell death types induce receptor shedding, but only apoptotic cells released nucleosome components. Conversely, necroptotic cells release lysosomal components by activating lysosomal exocytosis at early stages of necroptosis- induced membrane permeabilisation and show reduced release of conventionally secreted cytokines.
Project description:Spatial tissue proteomics integrating whole-slide imaging, laser microdissection and ultrasensitive mass spectrometry is a powerful approach to link cellular phenotypes to functional proteome states in (patho)physiology. To be applicable to large patient cohorts and low sample input amounts, including single-cell applications, loss-minimized and streamlined end-to-end workflows are key. We here introduce an automated sample preparation protocol for laser microdissected samples utilizing the cellenONE® robotic system, which has the capacity to process 192 samples in three hours. Following laser microdissection collection directly into the proteoCHIP LF 48 or EVO 96 chip, our optimized protocol facilitates lysis, formalin de-crosslinking and tryptic digest of low-input archival tissue samples. The seamless integration with the Evosep ONE LC system by centrifugation allows ‘on-the-fly’ sample clean-up, particularly pertinent for laser microdissected workflows. We validate our method in human tonsil archival tissue, where we profile proteomes of spatially-defined B-cell, T-cell and epithelial microregions of 4,000 µm2 to a depth of ~2,000 proteins and with high cell type specificity. We finally provide detailed equipment templates and experimental guidelines for broad accessibility.
Project description:Mutational signatures in esophageal squamous cell carcinoma from eight countries of varying incidence – sequence data (Mutographs)