Localization of protein aggregation in Escherichia coli is governed by diffusion and nucleoid macromolecular crowding effect

Aggregates of misfolded proteins are a hallmark of many age-related diseases. Recently, they have been linked to aging of Escherichia coli (E. coli) where protein aggregates accumulate at the old pole region of the aging bacterium. Because of the potential of E. coli as a model organism, elucidating aging and protein aggregation in this bacterium may pave the way to significant advances in our global understanding of aging. A first obstacle along this path is to decipher the mechanisms by which protein aggregates are targeted to specific intercellular locations. Here, using an integrated approach based on individual-based modeling, time-lapse fluorescence microscopy and automated image analysis, we show that the movement of aging-related protein aggregates in E. coli is purely diffusive (Brownian). Using single-particle tracking of protein aggregates in live E. coli cells, we estimated the average size and diffusion constant of the aggregates. Our results evidence that the aggregates passively diffuse within the cell, with diffusion constants that depend on their size in agreement with the Stokes-Einstein law. However, the aggregate displacements along the cell long axis are confined to a region that roughly corresponds to the nucleoid-free space in the cell pole, thus confirming the importance of increased macromolecular crowding in the nucleoids. We thus used 3d individual-based modeling to show that these three ingredients (diffusion, aggregation and diffusion hindrance in the nucleoids) are sufficient and necessary to reproduce the available experimental data on aggregate localization in the cells. Taken together, our results strongly support the hypothesis that the localization of aging-related protein aggregates in the poles of E. coli results from the coupling of passive diffusion- aggregation with spatially non-homogeneous macromolecular crowding. They further support the importance of "soft" intracellular structuring (based on macromolecular crowding) in diffusion-based protein localization in E. coli.Comment: PLoS Computational Biology (2013

Age at menarche in Canada: results from the National Longitudinal Survey of Children & Youth

<p>Abstract</p> <p>Background</p> <p>Given the downward trend in age at menarche and its implications for the reproductive health and wellbeing of women, little is known about menarcheal age in Canada. Most Canadian studies are only representative of specific populations. The present study, therefore, aims to assess the distribution of age at menarche for Canadian girls and explore its variation across socio-economic and demographic factors.</p> <p>Methods</p> <p>The analysis of the study was based on all female respondents aged 14 to 17 years during Cycle 4 (2000/2001) of the National Longitudinal Survey of Children & Youth (NLSCY). The main outcome was age at menarche assessed as the month and year of the occurrence of the first menstrual cycle. Kaplan Meier was used to estimate the mean and median of age at menarche. Chi-square test was used to assess the differences in early, average and later maturers across the different levels of socio-economic and demographic variables. Bootstrapping was performed to account for the complex sampling design.</p> <p>Results</p> <p>The total number of girls analyzed in this study was 1,403 weighted to represent 601,911 Canadian girls. The estimated mean and median of age at menarche was 12.72 years (standard deviation = 1.05) and 12.67 years, respectively. The proportions of early (< 11.53 years), average (≥11.53 years and ≤13.91 years) and late maturers (> 13.91 years) were 14.6% (95% confidence interval (CI): 11.92-17.35), 68.0% (95% CI: 63.82-72.17) and 17.4% (95% CI: 14.10-20.63), respectively. Variations across the menarcheal groups were statistically significant for the province of residence, household income and family type.</p> <p>Conclusion</p> <p>The findings of the study pave the way for future Canadian research. More studies are warranted to understand menarcheal age in terms of its variation across the provinces, the secular trend over time and its potential predictors.</p

Eddy-resolving simulation of plankton ecosystem dynamics in the California Current System

Author Posting. © Elsevier B.V., 2006. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Deep Sea Research Part I: Oceanographic Research Papers 53 (2006): 1483-1516, doi:10.1016/j.dsr.2006.06.005.We study the dynamics of the planktonic ecosystem in the coastal upwelling zone within the California Current System using a three-dimensional, eddy-resolving circulation model coupled to an ecosystem/biogeochemistry model. The physical model is based on the Regional Oceanic Modeling System (ROMS), configured at a resolution of 15 km for a domain covering the entire U.S. West Coast, with an embedded child grid covering the central California upwelling region at a resolution of 5 km. The model is forced with monthly mean boundary conditions at the open lateral boundaries as well as at the surface. The ecological/biogeochemical model is nitrogen based, includes single classes for phytoplankton and zooplankton, and considers two detrital pools with different sinking speeds. The model also explicitly simulates a variable chlorophyll-to-carbon ratio. Comparisons of model results with either remote sensing observations (AVHRR, SeaWiFS) or in situ measurements from the CalCOFI program indicate that our model is capable of replicating many of the large-scale, time averaged features of the coastal upwelling system. An exception is the underestimation of the chlorophyll levels in the northern part of the domain, perhaps because of the lack of short-term variations in the forcing from the atmosphere. Another shortcoming is that the modeled thermocline is too diffuse, and that the upward slope of the isolines toward the coast is too small. Detailed time-series comparisons with observations from Monterey Bay reveal similar agreements and discrepancies. We attribute the good agreement between the modeled and observed ecological properties in large part to the accuracy of the physical fields. In turn, many of the discrepancies can be traced back to our use of monthly mean forcing. Analysis of the ecosystem structure and dynamics reveal that the magnitude and pattern of phytoplankton biomass in the nearshore region are determined largely by the balance of growth and zooplankton grazing, while in the offshore region, growth is balanced by mortality. The latter appears to be inconsistent with in situ observations and is a result of our consideration of only one zooplankton size class (mesozooplankton), neglecting the importance of microzooplankton grazing in the offshore region. A comparison of the allocation of nitrogen into the different pools of the ecosystem in the 3-D results with those obtained from a box model configuration of the same ecosystem model reveals that only a few components of the ecosystem reach a local steady-state, i.e. where biological sources and sinks balance each other. The balances for the majority of the components are achieved by local biological source and sink terms balancing the net physical divergence, confirming the importance of the 3-D nature of circulation and mixing in a coastal upwelling system.Most of this work has been made possible by two grants from NASA. Additional support is acknowledged from NSF’s ITR program

Transit of H2O2 across the endoplasmic reticulum membrane is not sluggish

Cellular metabolism provides various sources of hydrogen peroxide (H2O2) in different organelles and compartments. The suitability of H2O2 as an intracellular signaling molecule therefore also depends on its ability to pass cellular membranes. The propensity of the membranous boundary of the endoplasmic reticulum (ER) to let pass H2O2 has been discussed controversially. In this essay, we challenge the recent proposal that the ER membrane constitutes a simple barrier for H2O2 diffusion and support earlier data showing that (i) ample H2O2 permeability of the ER membrane is a prerequisite for signal transduction, (ii) aquaporin channels are crucially involved in the facilitation of H2O2 permeation, and (iii) a proper experimental framework not prone to artifacts is necessary to further unravel the role of H2O2 permeation in signal transduction and organelle biology. © 2016 Elsevier Inc

Coastal and Inland Aquatic Data Products for the Hyperspectral Infrared Imager (HyspIRI)

The HyspIRI Aquatic Studies Group (HASG) has developed a conceptual list of data products for the HyspIRI mission to support aquatic remote sensing of coastal and inland waters. These data products were based on mission capabilities, characteristics, and expected performance. The topic of coastal and inland water remote sensing is very broad. Thus, this report focuses on aquatic data products to keep the scope of this document manageable. The HyspIRI mission requirements already include the global production of surface reflectance and temperature. Atmospheric correction and surface temperature algorithms, which are critical to aquatic remote sensing, are covered in other mission documents. Hence, these algorithms and their products were not evaluated in this report. In addition, terrestrial products (e.g., land use land cover, dune vegetation, and beach replenishment) were not considered. It is recognized that coastal studies are inherently interdisciplinary across aquatic and terrestrial disciplines. However, products supporting the latter are expected to already be evaluated by other components of the mission. The coastal and inland water data products that were identified by the HASG, covered six major environmental and ecological areas for scientific research and applications: wetlands, shoreline processes, the water surface, the water column, bathymetry and benthic cover types. Accordingly, each candidate product was evaluated for feasibility based on the HyspIRI mission characteristics and whether it was unique and relevant to the HyspIRI science objectives

Influence of plasma turbulence on microwave propagation

It is not fully understood how electromagnetic waves propagate through plasma density fluctuations when the size of the fluctuations is comparable with the wavelength of the incident radiation. In this paper, the perturbing effect of a turbulent plasma density layer on a traversing microwave beam is simulated with full-wave simulations. The deterioration of the microwave beam is calculated as a function of the characteristic turbulence structure size, the turbulence amplitude, the depth of the interaction zone and the size of the waist of the incident beam. The maximum scattering is observed for a structure size on the order of half the vacuum wavelength. The scattering and beam broadening was found to increase linearly with the depth of the turbulence layer and quadratically with the fluctuation strength. Consequences for experiments and 3D effects are considered.Comment: 16 pages, 13 figures. This is an author-created, un-copyedited version of an article submitted for publication in Plasma Physics and Controlled Fusion. IoP Publishing Ltd is not responsible for any errors or omissions in this version of the manuscript or any version derived from i

Physiologically relevant reconstitution of iron-sulfur cluster biosynthesis uncovers persulfide- processing functions of ferredoxin-2 and frataxin

Iron-sulfur (Fe-S) clusters are essential protein cofactors whose biosynthetic defects lead to severe diseases among which is Friedreich's ataxia caused by impaired expression of frataxin (FXN). Fe-S clusters are biosynthesized on the scaffold protein ISCU, with cysteine desulfurase NFS1 providing sulfur as persulfide and ferredoxin FDX2 supplying electrons, in a process stimulated by FXN but not clearly understood. Here, we report the breakdown of this process, made possible by removing a zinc ion in ISCU that hinders iron insertion and promotes non-physiological Fe-S cluster synthesis from free sulfide in vitro. By binding zinc-free ISCU, iron drives persulfide uptake from NFS1 and allows persulfide reduction into sulfide by FDX2, thereby coordinating sulfide production with its availability to generate Fe-S clusters. FXN stimulates the whole process by accelerating persulfide transfer. We propose that this reconstitution recapitulates physiological conditions which provides a model for Fe-S cluster biosynthesis, clarifies the roles of FDX2 and FXN and may help develop Friedreich's ataxia therapies

Disinfection of Ocular Cells and Tissues by Atmospheric-Pressure Cold Plasma

Background: Low temperature plasmas have been proposed in medicine as agents for tissue disinfection and have received increasing attention due to the frequency of bacterial resistance to antibiotics. This study explored whether atmospheric-pressure cold plasma (APCP) generated by a new portable device that ionizes a flow of helium gas can inactivate ocular pathogens without causing significant tissue damage. Methodology and Principal Findings: We tested the APCP effects on cultured Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Candida albicans, Aspergillus fumigatus and Herpes simplex virus-1, ocular cells (conjunctival fibroblasts and keratocytes) and ex-vivo corneas. Exposure to APCP for 0.5 to 5 minutes significantly reduced microbial viability (colony-forming units) but not human cell viability (MTT assay, FACS and Tunel analysis) or the number of HSV-1 plaque-forming units. Increased levels of intracellular reactive oxygen species (ROS) in exposed microorganisms and cells were found using a FACS-activated 2',7'-dichlorofluorescein diacetate probe. Immunoassays demonstrated no induction of thymine dimers in cell cultures and corneal tissues. A transient increased expression of 8-OHdG, genes and proteins related to oxidative stress (OGG1, GPX, NFE2L2) was determined in ocular cells and corneas by HPLC, qRT-PCR and Western blot analysis. Conclusions: A short application of APCP appears to be an efficient and rapid ocular disinfectant for bacteria and fungi without significant damage on ocular cells and tissues, although the treatment of conjunctival fibroblasts and keratocytes caused a time-restricted generation of intracellular ROS and oxidative stress-related responses