Evaluation of Glucose Uptake in Normal and Cancer Cell Lines by Positron Emission Tomography.

To date, there is no definitive demonstration of the utility of positron emission tomography (PET) in studying glucose metabolism in cultured cell lines. Thus, this study was designed to compare PET to more standardized methods for the quantitative assessment of glucose uptake in nontransformed and transformed living cells and to validate PET for metabolic studies in vitro. Human colon and breast carcinoma cell lines and mouse embryo fibroblasts were evaluated for [ 18 F]fluorodeoxyglucose ([ 18 F]FDG) uptake by PET and autoradiography and 2-deoxyglucose (2-DG) incorporation by colorimetric assay and analyzed for the radiotoxic effects of [ 18 F]FDG and the expression levels of glucose transporters. Indeed, [ 18 F]FDG incorporation on PET was comparable to [ 18 F]FDG uptake by autoradiography and 2-DG incorporation by colorimetric assay, although radiotracer-based methods exhibited more pronounced differences between individual cell lines. As expected, these data correlated with glucose transporters 1 to 4 and hexokinase II expression in tumor cell lines and mouse fibroblasts. Notably, [ 18 F]FDG incorporation resulted in low apoptotic rates, with fibroblasts being slightly more sensitive to radiotracer-induced cell death. The quantitative analysis of [ 18 F]FDG uptake in living cells by PET represents a valuable and reproducible method to study tumor cell metabolism in vitro, being representative of the differences in the molecular profile of normal and tumor cell lines

Ubiquitin, a central component of selective cytoplasmic proteolysis, is linked to proteins residing at the locus of non-selective proteolysis, the vacuole

Ubiquitin, an evolutionary highly conserved protein, is known to be involved in selective proteolysis in the cytoplasm. Here we show that ubiquitin-protein conjugates are also found in the yeast vacuole. Mutants defective in the major vacuolar endopeptidases, proteinase yscA and yscB, lead to accumulation of ubiquitin-protein conjugates in this cellular organelle.

Workshop on Assessing the Impact of Fishing on Oceanic Carbon (WKFISHCARBON; outputs from 2023 meeting)

Rapports Scientifiques du CIEM. Volume 6, nº 12The Workshop on Assessing the Impact of Fishing on Oceanic Carbon (WKFISHCARBON) was set up to provide ICES and stakeholders with a summary of knowledge on the role of fishing in the process of carbon budgets, sequestration and footprint in the ocean. The workshop addressed the potential impact of fishing on the biological carbon pump (BCP), the possible impacts of bottom trawling on carbon stores in the seabed, as well as considering emissions from fishing vessels. The overall aim was to generate proposals on how to develop an ICES approach to fishing and its role in the ocean carbon budget, and to develop a roadmap for a way forward. The main findings were that knowledge of the BCP in the open ocean was reasonably well developed, but that key gaps existed. In particular, information on the biomass of mesopelagic fish and other biota, and of some of the key processes e.g. fluxes and fish bioenergetics. Knowledge is much weaker for the BCP in shelf seas, where the bulk of fishing occurs. In particular, while biomass of fish was often well quantified, unlike the open ocean, the understanding of the important processes was lacking, particularly for the fate of faecal pellets and deadfall at the seabed. There is extensive scientific knowledge of the impact of fishing on the seabed, but what is un-clear is what it means for seabed carbon storage. There have been numbers of studies, which give a very divided view on this. There has also been open controversy about this in the literature. Physical disturbance to the seabed from fishing can affect sediment transport and has the potential to facilitate remineralization, but precise impacts will depend on habitat, fishing métier, and other environmental factors. From this, it is clear that more research is needed to resolve the controversy, and to quantify the impacts from different fishing gears and on different substrates or habitats in terms of carbon storage. There has been much more research on minimizing fuel use by fishing vessels, and hence emissions, but this has mainly focused on fuel efficiency, fuel use per unit of landed catch, and less on the total emissions. Baselines for fuel use are available at the global level, but are lacking at the national and vessel level. There is a need for standardization of methodologies and protocols, and for improving the uptake of fuel conservation measures by industry, as well as for improving the uptake of existing and potential fuel conservation and efficiency measures by industry. Finally, a roadmap was proposed to develop research and synthesis, on the understandings of the processes involved, the metrics and how to translate this into possible advice for policy-makers. To that end, a further workshop was proposed in 2024.info:eu-repo/semantics/publishedVersio

Allele-Specific HLA Loss and Immune Escape in Lung Cancer Evolution

Immune evasion is a hallmark of cancer. Losing the ability to present neoantigens through human leukocyte antigen (HLA) loss may facilitate immune evasion. However, the polymorphic nature of the locus has precluded accurate HLA copy-number analysis. Here, we present loss of heterozygosity in human leukocyte antigen (LOHHLA), a computational tool to determine HLA allele-specific copy number from sequencing data. Using LOHHLA, we find that HLA LOH occurs in 40% of non-small-cell lung cancers (NSCLCs) and is associated with a high subclonal neoantigen burden, APOBEC-mediated mutagenesis, upregulation of cytolytic activity, and PD-L1 positivity. The focal nature of HLA LOH alterations, their subclonal frequencies, enrichment in metastatic sites, and occurrence as parallel events suggests that HLA LOH is an immune escape mechanism that is subject to strong microenvironmental selection pressures later in tumor evolution. Characterizing HLA LOH with LOHHLA refines neoantigen prediction and may have implications for our understanding of resistance mechanisms and immunotherapeutic approaches targeting neoantigens. Video Abstract [Figure presented] Development of the bioinformatics tool LOHHLA allows precise measurement of allele-specific HLA copy number, improves the accuracy in neoantigen prediction, and uncovers insights into how immune escape contributes to tumor evolution in non-small-cell lung cancer

Fc-Optimized Anti-CD25 Depletes Tumor-Infiltrating Regulatory T Cells and Synergizes with PD-1 Blockade to Eradicate Established Tumors

CD25 is expressed at high levels on regulatory T (Treg) cells and was initially proposed as a target for cancer immunotherapy. However, anti-CD25 antibodies have displayed limited activity against established tumors. We demonstrated that CD25 expression is largely restricted to tumor-infiltrating Treg cells in mice and humans. While existing anti-CD25 antibodies were observed to deplete Treg cells in the periphery, upregulation of the inhibitory Fc gamma receptor (FcγR) IIb at the tumor site prevented intra-tumoral Treg cell depletion, which may underlie the lack of anti-tumor activity previously observed in pre-clinical models. Use of an anti-CD25 antibody with enhanced binding to activating FcγRs led to effective depletion of tumor-infiltrating Treg cells, increased effector to Treg cell ratios, and improved control of established tumors. Combination with anti-programmed cell death protein-1 antibodies promoted complete tumor rejection, demonstrating the relevance of CD25 as a therapeutic target and promising substrate for future combination approaches in immune-oncology

Phylogenetic ctDNA analysis depicts early-stage lung cancer evolution.

The early detection of relapse following primary surgery for non-small-cell lung cancer and the characterization of emerging subclones, which seed metastatic sites, might offer new therapeutic approaches for limiting tumour recurrence. The ability to track the evolutionary dynamics of early-stage lung cancer non-invasively in circulating tumour DNA (ctDNA) has not yet been demonstrated. Here we use a tumour-specific phylogenetic approach to profile the ctDNA of the first 100 TRACERx (Tracking Non-Small-Cell Lung Cancer Evolution Through Therapy (Rx)) study participants, including one patient who was also recruited to the PEACE (Posthumous Evaluation of Advanced Cancer Environment) post-mortem study. We identify independent predictors of ctDNA release and analyse the tumour-volume detection limit. Through blinded profiling of postoperative plasma, we observe evidence of adjuvant chemotherapy resistance and identify patients who are very likely to experience recurrence of their lung cancer. Finally, we show that phylogenetic ctDNA profiling tracks the subclonal nature of lung cancer relapse and metastasis, providing a new approach for ctDNA-driven therapeutic studies

An extended mtDNA phylogeography for the alpine newt illuminates the provenance of introduced populations

Many herpetofauna species have been introduced outside of their native range. MtDNA barcoding is regularly used to determine the provenance of such populations. The alpine newt has been introduced across the Netherlands, the United Kingdom and Ireland. However, geographical mtDNA structure across the natural range of the alpine newt is still incompletely understood and certain regions are severely undersampled. We collect mtDNA sequence data of over seven hundred individuals, from both the native and the introduced range. The main new insights from our extended mtDNA phylogeography are that 1) haplotypes from Spain do not form a reciprocally monophyletic clade, but are nested inside the mtDNA clade that covers western and eastern Europe; and 2) haplotypes from the northwest Balkans form a monophyletic clade together with those from the Southern Carpathians and Apuseni Mountains. We also home in on the regions where the distinct mtDNA clades meet in nature. We show that four out of the seven distinct mtDNA clades that comprise the alpine newt are implicated in the introductions in the Netherlands, United Kingdom and Ireland. In several introduced localities, two distinct mtDNA clades co-occur. As these mtDNA clades presumably represent cryptic species, we urge that the extent of genetic admixture between them is assessed from genome-wide nuclear DNA markers. We mobilized a large number of citizen scientists in this project to support the collection of DNA samples by skin swabbing and underscore the effectiveness of this sampling technique for mtDNA barcoding

Track E Implementation Science, Health Systems and Economics

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/138412/1/jia218443.pd

How are Australian higher education institutions contributing to change through innovative teaching and learning in virtual worlds?

Over the past decade, teaching and learning in virtual worlds has been at the forefront of many higher education institutions around the world. The DEHub Virtual Worlds Working Group (VWWG) consisting of Australian and New Zealand higher education academics was formed in 2009. These educators are investigating the role that virtual worlds play in the future of education and actively changing the direction of their own teaching practice and curricula. 47 academics reporting on 28 Australian higher education institutions present an overview of how they have changed directions through the effective use of virtual worlds for diverse teaching and learning activities such as business scenarios and virtual excursions, role-play simulations, experimentation and language development. The case studies offer insights into the ways in which institutions are continuing to change directions in their teaching to meet changing demands for innovative teaching, learning and research in virtual worlds. This paper highlights the ways in which the authors are using virtual worlds to create opportunities for rich, immersive and authentic activities that would be difficult or not possible to achieve through more traditional approaches