The modification of CD4+ T cells with exogenous nucleic acids or proteins is a critical step in several research and therapeutic applications, such as HIV studies and cancer immunotherapies. However, efficient cell transfections are not always easily achieved when working with these primary hard-to-transfect cells. While the modification of T cells is typically performed by viral transduction or electroporation, their use is associated with safety issues or cytotoxicity. Vapor nanobubble (VNB) photoporation with sensitizing gold nanoparticles (AuNPs) has recently emerged as a new technology for safe and flexible cell transfections. In this work, we evaluated the potential of VNB photoporation as a novel technique for the intracellular delivery of macromolecules in primary human CD4+ T cells using fluorescent dextrans as model molecules. Our results show that VNB photoporation enables efficient delivery of fluorescent dextrans of 10 kDa in Jurkat (>60% FD10+ cells) as well as in primary human CD4+ T cells (±40% FD10+ cells), with limited cell toxicity (>70% cell viability). We also demonstrated that the technique allows the delivery of dextrans that are up to 500 kDa in Jurkat cells, suggesting its applicability for the delivery of biological macromolecules with a wide range of molecular weights. Altogether, VNB photoporation represents a promising technique for the universal delivery of macromolecules in view of engineering CD4+ T cells for use in a wide variety of research and therapeutic applications.
the molecular, mesoscopic and microscopic structures of partially digested extruded maize starches varying in amylose content and digestion rate. Methods: Three maize starches with 27, 50 and 80% amylose levels were extruded at different moisture feed rates. Extrudates with varying moisture contents were digested (in vitro) at various times and undigested residues were freeze-dried. Raw, extrudate and partially digested starches were analysed for %starch digestibility, resistant starch, amylopectin branch length profile, helices, crystallinity, lamellar periodicity etc. Results: Digestibility data showed the normal starch was close to 100% digested, whereas HAMS were~30% digestible as granules and~75% in extrudates. There was a significant decrease in long amylopectin chains due to shear during extrusion, resulting in shorter chains influencing starch digestibility. NMR and crystallinity data showed that extended digestion of extrudate results in a significant increase in molecular order or helical content. Mechanism of enzyme resistance of granular high-amylose starches is qualitatively different to that for processed starches. Conclusions: Incorporation of HAMS to enhance resistant starch in foods can be achieved from either cooked or uncooked forms, but that the consequences may not be the same based on the different mechanisms involved in amylase digestion. Funding source(s): ARC. NEW STRAWBERRY BREEDING LINES e ENHANCED PHYTOCHEMICAL COMPOSITION AND BIOACCESSIBILITYBackground/Aims: The aim of this study was to investigate the effect of oat b-glucan (bG) on bile salts (BS) diffusion across the intestinal mucosa.Methods: Pigs used as a human model were fed a control diet (n ¼ 6) or a diet
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