Engineered mosaic protein polymers; a simple route to multifunctional biomaterials

Abstract: Background: Engineered living materials (ELMs) are an exciting new frontier, where living organisms create highly functional materials. In particular, protein ELMs have the advantage that their properties can be manipulated via simple molecular biology. Caf1 is a protein ELM that is especially attractive as a biomaterial on account of its unique combination of properties: bacterial cells export it as a massive, modular, non-covalent polymer which is resistant to thermal and chemical degradation and free from animal material. Moreover, it is biologically inert, allowing the bioactivity of each 15 kDa monomeric Caf1 subunit to be specifically engineered by mutagenesis and co-expressed in the same Escherichia coli cell to produce a mixture of bioactive Caf1 subunits. Results: Here, we show by gel electrophoresis and transmission electron microscopy that the bacterial cells combine these subunits into true mosaic heteropolymers. By combining two separate bioactive motifs in a single mosaic polymer we demonstrate its utility by stimulating the early stages of bone formation by primary human bone marrow stromal cells. Finally, using a synthetic biology approach, we engineer a mosaic of three components, demonstrating that Caf1 complexity depends solely upon the variety of monomers available. Conclusions: These results demonstrate the utility of engineered Caf1 mosaic polymers as a simple route towards the production of multifunctional biomaterials that will be useful in biomedical applications such as 3D tissue culture and wound healing. Additionally, in situ Caf1 producing cells could create complex bacterial communities for biotechnology. Graphical abstract

Bounding biomass in the Fisher equation

The FKPP equation with a variable growth rate and advection by an incompressible velocity field is considered as a model for plankton dispersed by ocean currents. If the average growth rate is negative then the model has a survival-extinction transition; the location of this transition in the parameter space is constrained using variational arguments and delimited by simulations. The statistical steady state reached when the system is in the survival region of parameter space is characterized by integral constraints and upper and lower bounds on the biomass and productivity that follow from variational arguments and direct inequalities. In the limit of zero-decorrelation time the velocity field is shown to act as Fickian diffusion with an eddy diffusivity much larger than the molecular diffusivity and this allows a one-dimensional model to predict the biomass, productivity and extinction transitions. All results are illustrated with a simple growth and stirring model.Comment: 32 Pages, 13 Figure

Dispersion in the open ocean seasonal pycnocline at scales of 1-10 km and 1-6 days

Author Posting. © American Meteorological Society, 2020. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 50(2), (2020): 415-437, doi:10.1175/JPO-D-19-0019.1.Results are presented from two dye release experiments conducted in the seasonal thermocline of the Sargasso Sea, one in a region of low horizontal strain rate (~10−6 s−1), the second in a region of intermediate horizontal strain rate (~10−5 s−1). Both experiments lasted ~6 days, covering spatial scales of 1–10 and 1–50 km for the low and intermediate strain rate regimes, respectively. Diapycnal diffusivities estimated from the two experiments were κz = (2–5) × 10−6 m2 s−1, while isopycnal diffusivities were κH = (0.2–3) m2 s−1, with the range in κH being less a reflection of site-to-site variability, and more due to uncertainties in the background strain rate acting on the patch combined with uncertain time dependence. The Site I (low strain) experiment exhibited minimal stretching, elongating to approximately 10 km over 6 days while maintaining a width of ~5 km, and with a notable vertical tilt in the meridional direction. By contrast, the Site II (intermediate strain) experiment exhibited significant stretching, elongating to more than 50 km in length and advecting more than 150 km while still maintaining a width of order 3–5 km. Early surveys from both experiments showed patchy distributions indicative of small-scale stirring at scales of order a few hundred meters. Later surveys show relatively smooth, coherent distributions with only occasional patchiness, suggestive of a diffusive rather than stirring process at the scales of the now larger patches. Together the two experiments provide important clues as to the rates and underlying processes driving diapycnal and isopycnal mixing at these scales.Results are presented from two dye release experiments conducted in the seasonal thermocline of the Sargasso Sea, one in a region of low horizontal strain rate (~10−6 s−1), the second in a region of intermediate horizontal strain rate (~10−5 s−1). Both experiments lasted ~6 days, covering spatial scales of 1–10 and 1–50 km for the low and intermediate strain rate regimes, respectively. Diapycnal diffusivities estimated from the two experiments were κz = (2–5) × 10−6 m2 s−1, while isopycnal diffusivities were κH = (0.2–3) m2 s−1, with the range in κH being less a reflection of site-to-site variability, and more due to uncertainties in the background strain rate acting on the patch combined with uncertain time dependence. The Site I (low strain) experiment exhibited minimal stretching, elongating to approximately 10 km over 6 days while maintaining a width of ~5 km, and with a notable vertical tilt in the meridional direction. By contrast, the Site II (intermediate strain) experiment exhibited significant stretching, elongating to more than 50 km in length and advecting more than 150 km while still maintaining a width of order 3–5 km. Early surveys from both experiments showed patchy distributions indicative of small-scale stirring at scales of order a few hundred meters. Later surveys show relatively smooth, coherent distributions with only occasional patchiness, suggestive of a diffusive rather than stirring process at the scales of the now larger patches. Together the two experiments provide important clues as to the rates and underlying processes driving diapycnal and isopycnal mixing at these scales.2020-08-0

Observations and numerical simulations of large-eddy circulation in the ocean surface mixed layer

Author Posting. © American Geophysical Union, 2014. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Geophysical Research Letters 41 (2014): 7584–7590, doi:10.1002/2014GL061637.Two near-surface dye releases were mapped on scales of minutes to hours temporally, meters to order 1 km horizontally, and 1–20 m vertically using a scanning, depth-resolving airborne lidar. In both cases, dye evolved into a series of rolls with their major axes approximately aligned with the wind and/or near-surface current. In both cases, roll spacing was also of order 5–10 times the mixed layer depth, considerably larger than the 1–2 aspect ratio expected for Langmuir cells. Numerical large-eddy simulations under similar forcing showed similar features, even without Stokes drift forcing. In one case, inertial shear driven by light winds induced large aspect ratio large-eddy circulation. In the second, a preexisting lateral mixed layer density gradient provided the dominant forcing. In both cases, the growth of the large-eddy structures and the strength of the resulting dispersion were highly dependent on the type of forcing.Support for the 2004 field experiment was provided by the Cecil H. and Ida M. Green Technology Innovation Fund and Coastal Ocean Institute grant 27001545, both through Woods Hole Oceanographic Institution, and by Office of Naval Research grant N00014-01-1-0984. Support for the 2011 field experiments was provided by ONR grants N00014-09-1-0194, N00014-09-1-0175, N00014-11-WX-21010, N00014-12-WX-21031, and N00014-09-1-0460 and NSF grants OCE-0751734 and OCE-0751653. Simulations were supported under grant N00014-09-1-0268.2015-05-0

Navigating Physicians’ Ethical and Legal Duties to Patients Seeking Unproven Interventions Abroad

Medical tourism (MT), the practice of traveling to another country to access medical care that is paid for out of pocket, has received considerable attention in the Canadian news media.Media and industry information sources, which are commonly accessed by medical tourists, might inadequately inform Canadians about MT safety concerns. As a result, there is concern among Canadian physicians and health and safety professionals that prospective medical tourists might not be well placed to make informed decisions about their care. As gatekeepers in the health care system and the first source of interaction between the health care system and patients, family physicians are well positioned to inform Canadians about these safety risks

Energetics of hydrogen impurities in aluminum and their effect on mechanical properties

The effects of hydrogen impurities in the bulk and on the surface of aluminum are theoretically investigated. Within the framework of density functional theory, we have obtained the dependence on H concentration of the stacking fault energy, the cleavage energy, the Al/H surface energy and the Al/H/Al interface formation energy. The results indicate a strong dependence of the slip energy barrier in the $[\bar 211]$ direction the cleavage energy in the [111] direction and the Al/H/Al interface formation energy, on H concentration and on tension. The dependence of the Al/H surface energy on H coverage is less pronounced, while the optimal H coverage is $\leq 0.25$ monolayer. The calculated activation energy for diffusion between high symmetry sites in the bulk and on the surface is practically the same, 0.167 eV. From these results, we draw conclusions about the possible effect of H impurities on mechanical properties, and in particular on their role in embrittlement of Al.Comment: 9 pages, 5 figure

An additional bolus of rapid-acting insulin to normalise postprandial cardiovascular risk factors following a high-carbohydrate high-fat meal in patients with type 1 diabetes: A randomised controlled trial.

AIM: To evaluate an additional rapid-acting insulin bolus on postprandial lipaemia, inflammation and pro-coagulation following high-carbohydrate high-fat feeding in people with type 1 diabetes. METHODS: A total of 10 males with type 1 diabetes [HbA1c 52.5 ± 5.9 mmol/mol (7.0% ± 0.5%)] underwent three conditions: (1) a low-fat (LF) meal with normal bolus insulin, (2), a high-fat (HF) meal with normal bolus insulin and (3) a high-fat meal with normal bolus insulin with an additional 30% insulin bolus administered 3-h post-meal (HFA). Meals had identical carbohydrate and protein content and bolus insulin dose determined by carbohydrate-counting. Blood was sampled periodically for 6-h post-meal and analysed for triglyceride, non-esterified-fatty acids, apolipoprotein B48, glucagon, tumour necrosis factor alpha, fibrinogen, human tissue factor activity and plasminogen activator inhibitor-1. Continuous glucose monitoring captured interstitial glucose responses. RESULTS: Triglyceride concentrations following LF remained similar to baseline, whereas triglyceride levels following HF were significantly greater throughout the 6-h observation period. The additional insulin bolus (HFA) normalised triglyceride similarly to low fat 3-6 h following the meal. HF was associated with late postprandial elevations in tumour necrosis factor alpha, whereas LF and HFA was not. Fibrinogen, plasminogen activator inhibitor-1 and tissue factor pathway levels were similar between conditions. CONCLUSION: Additional bolus insulin 3 h following a high-carbohydrate high-fat meal prevents late rises in postprandial triglycerides and tumour necrosis factor alpha, thus improving cardiovascular risk profile

Airfall on Comet 67P/Churyumov–Gerasimenko

We here study the transfer process of material from one hemisphere to the other (deposition of airfall material) on an active comet nucleus, specifically 67P/Churyumov–Gerasimenko. Our goals are to: 1) quantify the thickness of the airfall debris layers and how it depends on the location of the target area, 2) determine the amount of H₂O and CO₂ ice that are lost from icy dust assemblages of different sizes during transfer through the coma, and 3) estimate the relative amount of vapor loss in airfall material after deposition in order to understand what locations are expected to be more active than others on the following perihelion approach. We use various numerical simulations, that include orbit dynamics, thermophysics of the nucleus and of individual coma aggregates, coma gas kinetics and hydrodynamics, as well as dust dynamics due to gas drag, to address these questions. We find that the thickness of accumulated airfall material varies substantially with location, and typically is of the order 0.1–1 m. The airfall material preserves substantial amounts of water ice even in relatively small (cm–sized) coma aggregates after a rather long (12 h) residence in the coma. However, CO₂ is lost within a couple of hours even in relatively large (dm–sized) aggregates, and is not expected to be an important component in airfall deposits. We introduce reachability and survivability indices to measure the relative capacity of different regions to simultaneously collect airfall and to preserve its water ice until the next perihelion passage, thereby grading their potential of contributing to comet activity during the next perihelion passage

Airfall on Comet 67P/Churyumov–Gerasimenko

We here study the transfer process of material from one hemisphere to the other (deposition of airfall material) on an active comet nucleus, specifically 67P/Churyumov–Gerasimenko. Our goals are to: 1) quantify the thickness of the airfall debris layers and how it depends on the location of the target area, 2) determine the amount of H₂O and CO₂ ice that are lost from icy dust assemblages of different sizes during transfer through the coma, and 3) estimate the relative amount of vapor loss in airfall material after deposition in order to understand what locations are expected to be more active than others on the following perihelion approach. We use various numerical simulations, that include orbit dynamics, thermophysics of the nucleus and of individual coma aggregates, coma gas kinetics and hydrodynamics, as well as dust dynamics due to gas drag, to address these questions. We find that the thickness of accumulated airfall material varies substantially with location, and typically is of the order 0.1–1 m. The airfall material preserves substantial amounts of water ice even in relatively small (cm–sized) coma aggregates after a rather long (12 h) residence in the coma. However, CO₂ is lost within a couple of hours even in relatively large (dm–sized) aggregates, and is not expected to be an important component in airfall deposits. We introduce reachability and survivability indices to measure the relative capacity of different regions to simultaneously collect airfall and to preserve its water ice until the next perihelion passage, thereby grading their potential of contributing to comet activity during the next perihelion passage

Observations and numerical simulations of large-eddy circulation in the ocean surface mixed layer

Two near-surface dye releases were mapped on scales of minutes to hours temporally, meters to order 1 km horizontally, and 1–20 m vertically using a scanning, depth-resolving airborne lidar. In both cases, dye evolved into a series of rolls with their major axes approximately aligned with the wind and/or near-surface current. In both cases, roll spacing was also of order 5–10 times the mixed layer depth, considerably larger than the 1–2 aspect ratio expected for Langmuir cells. Numerical large-eddy simulations under similar forcing showed similar features, even without Stokes drift forcing. In one case, inertial shear driven by light winds induced large aspect ratio large-eddy circulation. In the second, a preexisting lateral mixed layer density gradient provided the dominant forcing. In both cases, the growth of the large-eddy structures and the strength of the resulting dispersion were highly dependent on the type of forcing.Keywords: lidar, large-eddy circulation, fluorescent dye, ocean surface mixed layer, numerical modelKeywords: lidar, large-eddy circulation, fluorescent dye, ocean surface mixed layer, numerical mode