9Poly lactide-co-glycolide (PLGA) is an important polymer matrix used to provide sustained 10 release across a range of active pharmaceutical ingredients (APIs) and works by hydrolytic 11 degradation within the body, thereby releasing entrapped drug. Processing and sterilisation 12 can impact on the morphology and chemistry of PLGA therefore influencing the hydrolysis 13 rate and in turn the release rate of any entrapped API. This paper has looked at the effect of 14 supercritical carbon dioxide (scCO 2 ) processing, gamma irradiation, comonomer ratio and 15 temperature on the hydrolysis of individual PLGA microparticles, using a combination of 16 Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) imaging, Scanning 17 Electron Microscopy (SEM), Differential Scanning Calorimetery (DSC) and Gel Permeation 18 chromatography (GPC) to facilitate a better understanding of the physiochemical factors 19affecting the hydrolysis rate. This work has shown that scCO 2 processing influences 20 hydrolysis rates by increasing the porosity of the PLGA microparticles, increasing the lactide 21 comonomer ratio decreases hydrolysis rates by reducing the hydrophilicity of the PLGA 22 microparticles and increasing the gamma irradiation dose systematically increases the rate of 23 hydrolysis due to reducing the overall molecular weight of the polymer matrix via a chain 24 scission mechanism. Moreover this work shows that ATR-FTIR imaging facilitates the 25 determination of a range of physicochemical parameters during the hydrolysis of a single 26 PLGA microparticle including water ingress, water/polymer interface dimensions, 27 degradation product distribution and hydrolysis rates for both lactide and glycolide 28 copolymer units from the same experiment. 29

For the first time, we report a series of time resolved images of a single PLGA microparticle undergoing hydrolysis at 70 °C that have been obtained using attenuated total reflectance-Fourier transform infrared spectroscopic (ATR-FTIR) imaging. A novel partially supervised non-linear curve fitting (NLCF) tool was developed to identify and fit peaks to the infrared spectrum obtained from each pixel within the 64 × 64 array. The output from the NLCF was evaluated by comparison with a traditional peak height (PH) data analysis approach and multivariate curve resolution alternating least squares (MCR-ALS) analysis for the same images, in order to understand the limitations and advantages of the NLCF methodology. The NLCF method was shown to facilitate consistent spatial resolution enhancement as defined using the step-edge approach on dry microparticle images when compared to images derived from both PH measurements and MCR-ALS. The NLCF method was shown to improve both the S/N and sharpness of images obtained during an evolving experiment, providing a better insight into the magnitude of hydration layers and particle dimension changes during hydrolysis. The NLCF approach facilitated the calculation of hydrolysis rate constants for both the glycolic (kG) and lactic (kL) acid segments of the PLGA copolymer. This represents a real advantage over MCR-ALS which could not distinguish between the two segments due to colinearity within the data. The NLCF approach made it possible to calculate the hydrolysis rate constants from a single pixel, unlike the peak height data analysis approach which suffered from poor S/N at each pixel. These findings show the potential value of applying NLCF to the study of real-time chemical processes at the micron scale, assisting in the understanding of the mechanisms of chemical processes that occur within microparticles and enhancing the value of the mid-IR ATR analysis.

Human rhinovirus (HRV), like coronavirus (HCoV), are positive-strand RNA viruses that cause both upper and lower respiratory tract illness, with their replication facilitated by concentrating RNA-synthesizing machinery in intracellular compartments made of modified host membranes, referred to as replication organelles (ROs). Here we report a non-canonical, essential function for stimulator of interferon genes (STING) during HRV infections. While the canonical function of STING is to detect cytosolic DNA and activate inflammatory responses, HRV infection triggers the release of STIM1-bound STING in the ER by lowering Ca2+, thereby allowing STING to interact with phosphatidylinositol 4-phosphate (PI4P) and traffic to ROs to facilitates viral replication and transmission via autophagy. Our results thus hint a critical function of STING in HRV viral replication and transmission, with possible implications for other RO-mediated RNA viruses.

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