Cancer cells are mechanically sensitive to physical properties of the microenvironment, which can affect downstream signaling to promote malignancy, in part through the modulation of metabolic pathways. Fluorescence Lifetime Imaging Microscopy (FLIM) can be used to measure the fluorescence lifetime of endogenous fluorophores, such as the metabolic co-factors NAD(P)H and FAD, in live samples. We used multiphoton FLIM to investigate the changes in cellular metabolism of 3D breast spheroids derived from MCF-10A and MD-MB-231 cell lines embedded in collagen with varying densities (1 vs. 4 mg/ml) over time (Day 0 vs. Day 3). MCF-10A spheroids demonstrated spatial gradients, with the cells closest to the spheroid edge exhibiting FLIM changes consistent with a shift towards oxidative phosphorylation (OXPHOS) while the spheroid core had changes consistent with a shift towards glycolysis. The MDA-MB-231 spheroids had a large shift consistent with increased OXPHOS with a more pronounced change at the higher collagen concentration. The MDA-MB-231 spheroids invaded into the collagen gel over time and cells that traveled the farthest had the largest changes consistent with a shift towards OXPHOS. Overall, these results suggest that the cells in contact with the extracellular matrix (ECM) and those that migrated the farthest had changes consistent with a metabolic shift towards OXPHOS. More generally, these results demonstrate the ability of multiphoton FLIM to characterize how spheroids metabolism and spatial metabolic gradients are modified by physical properties of the 3D ECM.

Morbidity from disease is increasingly being understood as dependent on wellbeing of the ecosystem, as opposed to an isolated element. However, the combined burden of disequilibrium of the gut microbiome ecosystem -which can facilitate proliferation of antimicrobial resistance (AMR) -is not well understood. The gap in knowledge grows when considering regions, such as the Gaza Strip, where the population continues to experience recurrent conflicts, movement restrictions, and high rates of poverty, food insecurity and unemployment. 1 The concurrent rise of gut microbiome disequilibrium, AMR proliferation, and displacement of populations globally -combined with lack of data -poses significant but unknown risk to public health, in displaced and host communities alike. Examining the current landscape of research, and existing gaps solidifies the need for interdisciplinary perspectives to address AMR through exploration of the gut microbiome. This holistic approach can help account for the political, socio-economic, and material factors, in the context of regions experiencing conflict/war. The growing role of microbiota in health may hold answers to questions surrounding understudied conditions such as AMR, and even non-communicable or chronic diseases.

Ensuring good storage practices (GSPs) of medicine outlets is important to maintain and ensure the safety, quality, and efficacy of dispensed medicines. The objective of this scoping review was to determine the most common GSP compliance issues within pharmacies and medicine outlets. PRISMA extension for scoping reviews guidelines were followed, and PubMed and Google Scholar were searched to identify the relevant primary literature from January 2016 and February 2022. All study designs in English were eligible for inclusion as long as they reported quantitative data for the fulfilment of individual GSP activities. Duplicate independent screening of the search results was undertaken. Quality assessment was performed on studies for final inclusion. Data extraction was performed by one reviewer, and the observed aggregate percentage compliance (% compliance), along with 95% confidence interval for the root GSP operation, was calculated. Of the 380 records identified, 15 articles were included. The studies were conducted in 10 countries and the majority were participatory research studies. Fire safety had the lowest aggregate % compliance (0.9%). Controlled substance, climate, light, ventilation and temperature and stock and bookkeeping operations all had overall aggregate % compliance levels below 50%. Despite evidence of basic controls and measures, we identified low compliance for many GSP operations. To ensure the integrity of medicines, it is important to increase the enforcement of security measures, invest resources in personnel training, account for informal vendor practices, and facilitate sharing global data publicly from external and governing body inspections and reviews of storage practices.

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