Submarine mass movements and their consequences

Submarine spreading is a type of mass movement that involves the extension and fracturing of a thin surficial layer of sediment into coherent blocks and their finite displacement on a gently sloping slip surface. Its characteristic seafloor signature is a repetitive pattern of parallel ridges and troughs oriented perpendicular to the direction of mass movement. We map ~30 km2 of submarine spreads on the upper slope of the Hikurangi margin, east of Poverty Bay, North Island, New Zealand, using multibeam echosounder and 2D multichannel seismic data. These data show that spreading occurs in thin, gently-dipping, parallel-bedded clay, silt and sandy sedimentary units deposited as lowstand clinoforms. More importantly, high-amplitude and reverse polarity seismic reflectors, which we interpret as evidence of shallow gas accumulations, occur extensively in the fine sediments of the upper continental slope, but are either significantly weaker or entirely absent where the spreads are located. We use this evidence to propose that shallow gas, through the generation of pore pressure, has played a key role in establishing the failure surface above which submarine spreading occurred. Additional dynamic changes in pore pressure could have been triggered by a drop in sea level during the Last Glacial Maximum and seismic loading.peer-reviewe

A 3‐D Model of Gas Generation, Migration, and Gas Hydrate Formation at a Young Convergent Margin (Hikurangi Margin, New Zealand)

We present a three-dimensional gas hydrate systems model of the southern Hikurangi subduction margin in eastern New Zealand. The model integrates thermal and microbial gas generation, migration, and hydrate formation. Modeling these processes has improved the understanding of factors controlling hydrate distribution. Three spatial trends of concentrated hydrate occurrence are predicted. The first trend (I) is aligned with the principal deformation front in the overriding Australian plate. Concentrated hydrate deposits are predicted at or near the apexes of anticlines and to be mainly sourced from focused migration and recycling of microbial gas generated beneath the hydrate stability zone. A second predicted trend (II) is related to deformation in the subducting Pacific plate associated with former Mesozoic subduction beneath Gondwana and the modern Pacific-Australian plate boundary. This trend is enhanced by increased advection of thermogenic gas through permeable layers in the subducting plate and focused migration into the Neogene basin fill above Cretaceous-Paleogene structures. The third trend (III) follows the northern margin of the Hikurangi Channel and is related to the presence of buried strata of the Hikurangi Channel system. The predicted trends are consistent with pronounced seismic reflection anomalies related to free gas in the pore space and strength of the bottom-simulating reflection. However, only trend I is also associated with clear and widespread seismic indications of concentrated gas hydrate. Total predicted hydrate masses at the southern Hikurangi Margin are between 52,800 and 69,800 Mt. This equates to 3.4–4.5 Mt hydrate/km2, containing 6.33 × 108–8.38 × 108 m3/km2 of methane

Distance dependence of photoinduced long-range electron transfer in zinc/ruthenium-modified myoglobins

An experimental investigation of the distance dependence of long-range electron transfer in zinc/ruthenium-modified myoglobins has been performed. The modified proteins were prepared by substitution of zinc mesoporphyrin IX diacid (ZnP) for the heme in each of four previously characterized pentaammineruthenium(III) (a_5Ru;a = NH_3) derivatives of sperm whale myoglobin (Mb): a_5Ru(His-48)Mb, a_5Ru(His-12)Mb, a_5Ru(His-116)Mb, a_5Ru(His-81)Mb. Electron transfer from the ZnP triplet excited state (^3ZnP*) to Ru^3+, ^3ZnP*-Ru^3+ → ZnP^+-Ru^2+ (ΔE° ~ 0.8V) was measured by time-resolved transient absorption spectroscopy: rate constants (k_f) are 7.0 × 10^4 (His-48), 1.0 × 10^2 (His-12), 8.9 × 10^1 (His-116), and 8.5 × 10^1 (His-81) s^-1 at 25 °C. Activation enthalpies calculated from the temperature dependences of the electron-transfer rates over the range 5-40 °C are 1.7 ± 1.6 (His-48), 4.7 ± 0.9 (His-12), 5.4 ± 0.4 (His-116), and 5.6 ± 2.5 (His-81) kcal mol^-1. Electron-transfer distances (d = closest ZnP edge to a_5Ru(His) edge; angstroms) were calculated to fall in the following ranges: His-48, 11.8-16.6; His-12, 21.5-22.3; His-116, 19.8-20.4; His-81, 18.8-19.3. The rate-distance equation is k_f = 7.8 × 10^8 exp[-0.9l(d - 3)] s^-1 . The data indicate that the ^3ZnP*-Ru(His-12)^3+ electronic coupling may be enhanced by an intervening tryptophan (Trp-14)

Interplay of septin amphipathic helices in sensing membrane-curvature and filament bundling

© The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Woods, B. L., Cannon, K. S., Vogt, E. J. D., Crutchley, J. M., & Gladfelter, A. S. Interplay of septin amphipathic helices in sensing membrane-curvature and filament bundling. Molecular Biology of the Cell, 32(20), (2021): mbcE20050303, https://doi.org/10.1091/mbc.E20-05-0303.The curvature of the membrane defines cell shape. Septins are GTP-binding proteins that assemble into heteromeric complexes and polymerize into filaments at areas of micron-scale membrane curvature. An amphipathic helix (AH) domain within the septin complex is necessary and sufficient for septins to preferentially assemble onto micron-scale curvature. Here we report that the nonessential fungal septin, Shs1, also has an AH domain capable of recognizing membrane curvature. In a septin mutant strain lacking a fully functional Cdc12 AH domain (cdc12-6), the C-terminal extension of Shs1, containing an AH domain, becomes essential. Additionally, we find that the Cdc12 AH domain is important for regulating septin filament bundling, suggesting septin AH domains have multiple, distinct functions and that bundling and membrane binding may be coordinately controlled.This work was supported by National Institutes of Health (NIH) Grant no. R01GM-130934 to A.S.G. B.L.W. was supported by the NIH Training Grant no. 2T32AI052080-16. K.S.C. and E.J.D.V. were supported in part by a grant from the National Institute of General Medical Sciences under award T32 GM119999

Morphology and Oxygen Sensor Response of Luminescent Ir-Labeled Poly(dimethylsiloxane)/Polystyrene Polymer Blend Films

Polymer films consisting of a linear poly(dimethylsiloxane) end-functionalized with a luminescent Ir(III) complex (Ir−PDMS), blended with polystyrene (PS), function as optical oxygen sensors. The sensor response arises by quenching of the luminescence from the Ir(III) chromophore by oxygen that permeates into the polymer film. The morphology and luminescence oxygen sensor properties of blend films consisting of Ir−PDMS and PS have been characterized by fluorescence microscopy, atomic force microscopy, and scanning electron microscopy. The investigations demonstrate that microscale phase segregation occurs in the films. In blends that contain a relatively small amount of Ir−PDMS in PS (ca. 10 wt %), the Ir−PDMS exists as circular domains, with diameters ranging from 2 to 5 μm, surrounded by the majority PS phase. For larger weight fractions of Ir−PDMS in the blends, the film morphology becomes bicontinuous. A novel epifluorescence microscopy method is applied that allows the construction of Stern−Volmer quenching images that quantify the oxygen sensor response of the blend films with micrometer spatial resolution. These images provide a map of the oxygen permeability of the polymer blend films with a spatial resolution of ca. 1 μm. The results of this investigation show that the micrometer-sized Ir−PMDS domains display a 2−3-fold higher oxygen sensor response compared to the surrounding PS matrix. This result is consistent with the fact that PDMS is considerably more gas permeable compared to PS. The relationship of the microscale morphology of the blends to their performance as macroscale optical oxygen sensors is discussed

Development and validation of broad-spectrum magnetic particle labelling processes for cell therapy manufacturing

Background Stem cells are increasingly seen as a solution for many health challenges for an ageing population. However, their potential benefits in the clinic are currently curtailed by technical challenges such as high cell dose requirements and point of care delivery, which pose sourcing and logistics challenges. Cell manufacturing solutions are currently in development to address the supply issue, and ancillary technologies such as nanoparticle-based labelling are being developed to improve stem cell delivery and enable post-treatment follow-up. Methods The application of magnetic particle (MP) labelling to potentially scalable cell manufacturing processes was investigated in a range of therapeutically relevant cells, including mesenchymal stromal cells (MSC), cardiomyocytes (CMC) and neural progenitor cells (ReN). The efficiency and the biological effect of particle labelling were analysed using fluorescent imaging and cellular assays. Results Flow cytometry and fluorescent microscopy confirmed efficient labelling of monolayer cultures. Viability was shown to be retained post labelling for all three cell types. MSC and CMC demonstrated higher tolerance to MP doses up to 100× the standard concentration. This approach was also successful for MP labelling of suspension cultures, demonstrating efficient MP uptake within 3 h, while cell viability was unaffected by this suspension labelling process. Furthermore, a procedure to enable the storing of MP-labelled cell populations to facilitate cold chain transport to the site of clinical use was investigated. When MP-labelled cells were stored in hypothermic conditions using HypoThermosol solution for 24 h, cell viability and differentiation potential were retained post storage for ReN, MSC and beating CMC. Conclusions Our results show that a generic MP labelling strategy was successfully developed for a range of clinically relevant cell populations, in both monolayer and suspension cultures. MP-labelled cell populations were able to undergo transient low-temperature storage whilst maintaining functional capacity in vitro. These results suggest that this MP labelling approach can be integrated into cell manufacturing and cold chain transport processes required for future cell therapy approaches

Development and validation of broad-spectrum magnetic particle labelling processes for cell therapy manufacturing

BACKGROUND: Stem cells are increasingly seen as a solution for many health challenges for an ageing population. However, their potential benefits in the clinic are currently curtailed by technical challenges such as high cell dose requirements and point of care delivery, which pose sourcing and logistics challenges. Cell manufacturing solutions are currently in development to address the supply issue, and ancillary technologies such as nanoparticle-based labelling are being developed to improve stem cell delivery and enable post-treatment follow-up. METHODS: The application of magnetic particle (MP) labelling to potentially scalable cell manufacturing processes was investigated in a range of therapeutically relevant cells, including mesenchymal stromal cells (MSC), cardiomyocytes (CMC) and neural progenitor cells (ReN). The efficiency and the biological effect of particle labelling were analysed using fluorescent imaging and cellular assays. RESULTS: Flow cytometry and fluorescent microscopy confirmed efficient labelling of monolayer cultures. Viability was shown to be retained post labelling for all three cell types. MSC and CMC demonstrated higher tolerance to MP doses up to 100× the standard concentration. This approach was also successful for MP labelling of suspension cultures, demonstrating efficient MP uptake within 3 h, while cell viability was unaffected by this suspension labelling process. Furthermore, a procedure to enable the storing of MP-labelled cell populations to facilitate cold chain transport to the site of clinical use was investigated. When MP-labelled cells were stored in hypothermic conditions using HypoThermosol solution for 24 h, cell viability and differentiation potential were retained post storage for ReN, MSC and beating CMC. CONCLUSIONS: Our results show that a generic MP labelling strategy was successfully developed for a range of clinically relevant cell populations, in both monolayer and suspension cultures. MP-labelled cell populations were able to undergo transient low-temperature storage whilst maintaining functional capacity in vitro. These results suggest that this MP labelling approach can be integrated into cell manufacturing and cold chain transport processes required for future cell therapy approaches

Spatial heterogeneity of the cytosol revealed by machine learning-based 3D particle tracking

© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in McLaughlin, G. A., Langdon, E. M., Crutchley, J. M., Holt, L. J., Forest, M. G., Newby, J. M., & Gladfelter, A. S. (2020). Spatial heterogeneity of the cytosol revealed by machine learning-based 3D particle tracking. Molecular Biology of the Cell, 31(14), 1498-1511, doi:10.1091/mbc.E20-03-0210.The spatial structure and physical properties of the cytosol are not well understood. Measurements of the material state of the cytosol are challenging due to its spatial and temporal heterogeneity. Recent development of genetically encoded multimeric nanoparticles (GEMs) has opened up study of the cytosol at the length scales of multiprotein complexes (20-60 nm). We developed an image analysis pipeline for 3D imaging of GEMs in the context of large, multinucleate fungi where there is evidence of functional compartmentalization of the cytosol for both the nuclear division cycle and branching. We applied a neural network to track particles in 3D and then created quantitative visualizations of spatially varying diffusivity. Using this pipeline to analyze spatial diffusivity patterns, we found that there is substantial variability in the properties of the cytosol. We detected zones where GEMs display especially low diffusivity at hyphal tips and near some nuclei, showing that the physical state of the cytosol varies spatially within a single cell. Additionally, we observed significant cell-to-cell variability in the average diffusivity of GEMs. Thus, the physical properties of the cytosol vary substantially in time and space and can be a source of heterogeneity within individual cells and across populations.We would like to thank the 2016 Physiology course and Christina Termini at the Marine Biological Laboratory in Woods Hole, MA, Gregory Brittingham, and Marcus Roper for initial experiments and perspectives on pipeline. We thank David Adalsteinsson for help with DataTank software and many conversations about image analysis on large datasets. We thank Emmanual Levy (Weizmann Institute) for providing plasmids encoding synthetic phase separating peptides. This work was supported by Google Cloud, the National Science Foundation (NSF), the National Institutes of Health (NIH), and the Natural Sciences and Engineering Research Council of Canada (NSERC). ASG, EML, and GAM were supported by the NSF (RoLs: 1840273), HHMI faculty scholar award and the NIH (R01GM081506). JMN was supported by the NSERC (RGPIN-2019-06435, RGPAS-2019-00014, DGECR-2019-00321) and the NSF (DMS-171474). MGF was supported by the NSF (DMS-1816630, DMS-1664645). LJH was supported by the NIH (R01GM132447)