Project description:Despite the recent progress, current treatment modalities are not able to eradicate cancer. We show that Microbeam Radiotherapy (MRT), an innovative type of Spatially Fractionated Radiotherapy, can control murine melanoma by activating the host’s own immune system. The beneficial effects are very pronounced in comparison to uniform radiotherapy, traditionally employed in the clinic. Our results displayed that MRT increased antigen presentation, activating Cytotoxic T Lymphocytes (CTLs) which are essential to MRT’s treatment efficacy in melanoma. Depletion of CTLs abrogated treatment response. Multiplex nucleic acid hybridization technology revealed key features of lymphocyte populations such as proliferation, differentiation, and ligand-receptor interactions. In addition, CTLs were shown to be essential for locoregional metastatic control and systemic abscopal effects confirmed by activation of antigen presenting cells and CTL trafficking in the tumour-draining lymph nodes. MRT induces a robust antitumour immune response, matching the characteristics of in situ vaccination, that could be exploited to treat a variety of treatment-resistant malignancies.
Project description:In this study, we design ultra-broadband optical absorbers, ultra-narrow optical waveguides, and ultra-small optical cavities comprising two-dimensional metallic photonic crystals that tolerate fabrication imperfections such as position and radius disorderings. The absorbers containing gold rods show an absorption amplitude of more than 90% under 54% position disordering at 200<λ<530 nm. The absorbers containing silver rods show an absorptance of more than 90% under 54% position disordering at 200<λ<400 nm. B-type straight waveguides that contain four rows of silver rods exposed to air reveal normalized transmittances of 75% and 76% under 32% position and 60% radius disorderings, respectively. B-type L-shaped waveguides containing four rows of silver rods show 76% and 90% normalized transmittances under 32% position and 40% radius disorderings, respectively. B-type cavities containing two rings of silver rods reveal 70% and 80% normalized quality factors under 32% position and 60% radius disorderings, respectively.
Project description:Mass spectrometry (MS)-based proteomics aims to characterize comprehensive proteomes in a fast and reproducible manner. Here, we present an ultra-fast scanning data-independent acquisition (DIA) strategy consisting on 2-Th precursor isolation windows, dissolving the differences between data-dependent and independent methods. This is achieved by pairing a Quadrupole Orbitrap mass spectrometer with the asymmetric track lossless (Astral) analyzer that provides >200 Hz MS/MS scanning speed, high resolving power and sensitivity, as well as low ppm-mass accuracy. Narrow window DIA enables profiling of up to 100 full yeast proteomes per day, or ~10,000 human proteins in half-an-hour. Moreover, multi-shot acquisition of fractionated samples allows comprehensive coverage of human proteomes in ~3h, showing comparable depth to next-generation RNA sequencing and with 10x higher throughput compared to current state-of-the-art MS. High quantitative precision and accuracy is demonstrated with high peptide coverage in a 3-species proteome mixture, quantifying 14,000+ proteins in a single run in half-an-hour.
Project description:Mass spectrometry (MS)-based proteomics aims to characterize comprehensive proteomes in a fast and reproducible manner. Here, we present an ultra-fast scanning data-independent acquisition (DIA) strategy consisting on 2-Th precursor isolation windows, dissolving the differences between data-dependent and independent methods. This is achieved by pairing a Quadrupole Orbitrap mass spectrometer with the asymmetric track lossless (Astral) analyzer that provides >200 Hz MS/MS scanning speed, high resolving power and sensitivity, as well as low ppm-mass accuracy. Narrow window DIA enables profiling of up to 100 full yeast proteomes per day, or ~10,000 human proteins in half-an-hour. Moreover, multi-shot acquisition of fractionated samples allows comprehensive coverage of human proteomes in ~3h, showing comparable depth to next-generation RNA sequencing and with 10x higher throughput compared to current state-of-the-art MS. High quantitative precision and accuracy is demonstrated with high peptide coverage in a 3-species proteome mixture, quantifying 14,000+ proteins in a single run in half-an-hour.
Project description:In this work, we pioneered a combination of ultra-low flow (ULF) high-efficiency ultra-narrow bore monolithic LC columns coupled to MS via a high-field asymmetric waveform ion mobility spectrometry (FAIMS) interface to evaluate the potential applicability for high sensitivity, robust and reproducible proteomic profiling of low ng-level complex biological samples.
Project description:These experiments were undertaken with the goal of identifying genes whose expression is enriched in or restricted to the sensory rays of the C. elegans male tail. We constructed two mutant strains in which ray development is either compromised (EM672) or enhanced (EM673), and harvested mRNA from adult males. Labeled cDNAs were compared on seven arrays (representing three different sets of mRNA preps). In all experiments, Channel 1 (green) represents the EM672 expression profile and Channel 2 (Red) corresponds to EM673. Ray-enriched genes would therefore generally be expected to have higher intensities in Channel 2 than in Channel 1. Experimental details and results from these studies are available in D.S. Portman and S.W. Emmons (2004) Identification of C. elegans sensory ray genes using whole-genome expression profiling. Groups of assays that are related as part of a time series. Computed
Project description:These experiments were undertaken with the goal of identifying genes whose expression is enriched in or restricted to the sensory rays of the C. elegans male tail. We constructed two mutant strains in which ray development is either compromised (EM672) or enhanced (EM673), and harvested mRNA from adult males. Labeled cDNAs were compared on seven arrays (representing three different sets of mRNA preps). In all experiments, Channel 1 (green) represents the EM672 expression profile and Channel 2 (Red) corresponds to EM673. Ray-enriched genes would therefore generally be expected to have higher intensities in Channel 2 than in Channel 1. Experimental details and results from these studies are available in D.S. Portman and S.W. Emmons (2004) Identification of C. elegans sensory ray genes using whole-genome expression profiling. Groups of assays that are related as part of a time series. Keywords: time_series_design