Nanoparticle-sensitized photoporation is an upcoming approach for intracellular delivery of biologics, combining high efficiency and throughput with excellent cell viability. However, as it relies on close contact between nanoparticles and cells, its translation towards clinical applications is hampered by safety and regulatory concerns. Here, we show that light-sensitive iron oxide nanoparticles (IONPs) embedded in biocompatible electrospun nanofibers induce membrane permeabilization by photothermal effects without direct cellular contact with IONPs. The photothermal nanofibers are successfully used to deliver effector molecules, including CRISPR/Cas9 ribonucleoprotein complexes and siRNA, in adherent and suspension cells, including embryonic stem cells and hard-to-transfect T-cells without affecting cell proliferation or phenotype. In vivo experiments furthermore demonstrate successful tumor regression in mice treated with CAR-T cells in which expression of PD1 is downregulated after nanofiber photoporation with siPD1. In conclusion, cell membrane permeabilization with photothermal nanofibers is a promising concept towards the safe and more efficient production of engineered cells for therapeutic applications, including stem cell or adoptive T cell therapy.

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Lithium isotope ratios play a key role in tracing weathering processes within the context of paleoclimatologic research. Therefore, accurate and precise determination of the isotopic composition of Li is required for a large variety of complex geological samples with widely different Li contents. This technical note describes the extension and fine-tuning of the existing cation exchange chromatographic procedures for isolation of Li to whole-rock carbonate and clay matrices prior to its isotopic analysis via multi-collector ICP-mass spectrometry. The method developed permits the isolation of >= 20 ng amounts of Li from carbonates and clays, by using a 25 cm long cylindrical column loaded with 8 and 2 mL of AG-50W-X8 cation exchange resin, respectively. A 0.5 M HCl solution is used as eluent. Li recoveries >= 99% were achieved and isotopic analysis using 10-20 ng of isolated Li resulted in accurate and precise delta Li-7 values - external precision (2SE) of 0.2 parts per thousand (N = 15). The analytical procedures developed allow Li isotope ratios to be investigated in parallel in both the clay and carbonate fractions of a geological sample within the context of paleoclimatological research

Keywords: methane seeps ROV Hikurangi Margin sea-floor observations temperature measurements bubble-release activity During the 'New Vents' SO191 cruise in 2007, the activity and distribution of seep sites on the gas-hydratebearing Hikurangi Margin, off northeastern New Zealand, were subjected to a highly detailed interdisciplinary study. Here we report on the visual observations and in situ measurements of physical properties performed with a ROV (remotely operated vehicle) and other video-guided platforms at two seep sites in the Rock Garden area; Faure Site and LM-3. The ROV allowed first ever visual observations of bubblereleasing methane seeps at the Hikurangi Margin. At Faure Site, bubble release was monitored during 4 dives, up to periods of 20 min. During the first dive, this resulted in the observation of six violent outbursts, each lasting 1 min over a three minute interval. These outbursts were accompanied by the displacement and resuspension of sediment grains, and the formation of small depressions, with a maximum diameter of 50 cm and depth of 15 cm, showing what is possibly an initial stage of pockmark formation. During subsequent dives at this bubble site, bubble release rates were rather constant and the previously observed outbursts could no longer be witnessed. At LM-3, the strongest manifestation of seep activity was a large platform (100 m 2 ), consisting of fresh authigenic carbonates, which was covered by seep fauna (live Bathymodiolus sp. mussels, Calyptogena sp. shells and live Lamellibrachia sp. tubeworms). Bubble activity near this platform was less prominent than at Faure Site. Our observations suggest that the two seep environments result from different types of methane release; mainly by bubble release at Faure Site and rather diffusive at LM-3. We propose a conceptual model where the different ways of methane release and seep environments may be explained by the depth of underlying hydrate occurrences and different tectonic histories of both seep sites.

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