Space- and time-resolved investigation on diffusion kinetics of human skin following macromolecule delivery by microneedle arrays

Microscale medical devices are being developed for targeted skin delivery of vaccines and the extraction of biomarkers, with the potential to revolutionise healthcare in both developing and developed countries. The effective clinical development of these devices is dependent on understanding the macro-molecular diffusion properties of skin. We hypothesised that diffusion varied according to specific skin layers. Using three different molecular weights of rhodamine dextran (RD) (MW of 70, 500 and 2000 kDa) relevant to the vaccine and therapeutic scales, we deposited molecules to a range of depths (0–300 µm) in ex vivo human skin using the Nanopatch device. We observed significant dissipation of RD as diffusion with 70 and 500 kDa within the 30 min timeframe, which varied with MW and skin layer. Using multiphoton microscopy, image analysis and a Fick’s law analysis with 2D cartesian and axisymmetric cylindrical coordinates, we reported experimental trends of epidermal and dermal diffusivity values ranging from 1–8 µm2 s-1 to 1–20 µm2 s-1 respectively, with a significant decrease in the dermal-epidermal junction of 0.7–3 µm2 s-1. In breaching the stratum corneum (SC) and dermal-epidermal junction barriers, we have demonstrated practical application, delivery and targeting of macromolecules to both epidermal and dermal antigen presenting cells, providing a sound knowledge base for future development of skin-targeting clinical technologies in humans

Mouse Acetylcholinesterase Enhances Neurite Outgrowth of Rat R28 Cells Through Interaction With Laminin-1

The enzyme acetylcholinesterase (AChE) terminates synaptic transmission at cholinergic synapses by hydrolyzing the neurotransmitter acetylcholine, but can also exert ‘non-classical’, morpho-regulatory effects on developing neurons such as stimulation of neurite outgrowth. Here, we investigated the role of AChE binding to laminin-1 on the regulation of neurite outgrowth by using cell culture, immunocytochemistry, and molecular biological approaches. To explore the role of AChE, we examined fiber growth of cells overexpressing different forms of AChE, and/or during their growth on laminin-1. A significant increase of neuritic growth as compared with controls was observed for neurons over-expressing AChE. Accordingly, addition of globular AChE to the medium increased total length of neurites. Co-transfection with PRIMA, a membrane anchor of AChE, led to an increase in fiber length similar to AChE overexpressing cells. Transfection with an AChE mutant that leads to the retention of AChE within cells had no stimulatory effect on neurite length. Noticeably, the longest neurites were produced by neurons overexpressing AChE and growing on laminin-1, suggesting that the AChE/laminin interaction is involved in regulating neurite outgrowth. Our findings demonstrate that binding of AChE to laminin-1 alters AChE activity and leads to increased neurite growth in culture. A possible mechanism of the AChE effect on neurite outgrowth is proposed due to the interaction of AChE with laminin-1

Washing and caustic leaching of Hanford tank sludge: Results of FY 1997 studies

The current plan for remediating the Hanford tank farms consists of waste retrieval, pretreatment, treatment (immobilization), and disposal. The tank wastes will be partitioned into high-level and low-level fractions. The HLW will be immobilized in a borosilicate glass matrix; the resulting glass canisters will then be disposed of in a geologic repository. Because of the expected high cost of HLW vitrification and geologic disposal, pretreatment processes will be implemented to reduce the volume of immobilized high-level waste (IHLW). Caustic leaching (sometimes referred to as enhanced sludge washing or ESW) represents the baseline method for pretreating Hanford tank sludges. Caustic leaching is expected to remove a large fraction of the Al, which is present in large quantities in Hanford tank sludges. A significant portion of the P is also expected to be removed from the sludge by metathesis of water-insoluble metal phosphates to insoluble hydroxides and soluble Na{sub 3}PO{sub 4}. Similar metathesis reactions can occur for insoluble sulfate salts, allowing the removal of sulfate from the HLW stream. This report describes the sludge washing and caustic leaching tests performed at the Pacific Northwest National Laboratory in FY 1996. The sludges used in this study were taken from Hanford tanks AN-104, BY-108, S-101, and S-111