Intra-colony channel morphology in Escherichia coli biofilms is governed by nutrient availability and substrate stiffness

Nutrient-transporting channels are found throughout mature Escherichia coli biofilms, however the influence of environmental conditions on intra-colony channel formation is poorly understood. We report the effect of different substrate nutrient concentrations and agar stiffness on the structure and distribution of intra-colony channels in mature E. coli colony biofilms using fluorescence mesoscopy and quantitative image analysis. Intra-colony channel width was observed to increase non-linearly with radial distance from the centre of the biofilm and channels were, on average, 50% wider at the centre of carbon-limited biofilms compared to nitrogen-limited biofilms. Channel density also differed in colonies grown on rich and minimal medium substrates, with the former creating a network of tightly packed channels and the latter leading to well-separated, wider channels with easily identifiable edges. We conclude that intra-colony channel morphology in E. coli biofilms is influenced by both substrate composition and nutrient availability.

Solution Structures of Bacillus anthracis Protective Antigen Proteins Using Small Angle Neutron Scattering and Protective Antigen 63 Ion Channel Formation Kinetics

We are studying the structures of bacterial toxins that form ion channels and enable macromolecule transport across membranes. For example, the crystal structure of the Staphylococcus aureus α-hemolysin (α-HL) channel in its functional state was confirmed using neutron reflectometry (NR) with the protein reconstituted in membranes tethered to a solid support. This method, which provides sub-nanometer structural information, could also test putative structures of the Bacillus anthracis protective antigen 63 (PA63) channel, locate where B. anthracis lethal factor and edema factor toxins (LF and EF, respectively) bind to it, and determine how certain small molecules can inhibit the interaction of LF and EF with the channel. We report here the solution structures of channel-forming PA63 and its precursor PA83 (which does not form channels) obtained with small angle neutron scattering. At near neutral pH, PA83 is a monomer and PA63 a heptamer. The latter is compared to two cryo-electron microscopy structures. We also show that although the α-HL and PA63 channels have similar structural features, unlike α-HL, PA63 channel formation in lipid bilayer membranes ceases within minutes of protein addition, which currently precludes the use of NR for elucidating the interactions between PA63, LF, EF, and potential therapeutic agents.

Defining the molecular mechanisms of the mitochondrial permeability transition through genetic manipulation of F-ATP synthase

F-ATP synthase is a leading candidate as the mitochondrial permeability transition pore (PTP) but the mechanism(s) leading to channel formation remain undefined. Here, to shed light on the structural requirements for PTP formation, we test cells ablated for g, OSCP and b subunits, and ρ0 cells lacking subunits a and A6L. Δg cells (that also lack subunit e) do not show PTP channel opening in intact cells or patch-clamped mitoplasts unless atractylate is added. Δb and ΔOSCP cells display currents insensitive to cyclosporin A but inhibited by bongkrekate, suggesting that the adenine nucleotide translocator (ANT) can contribute to channel formation in the absence of an assembled F-ATP synthase. Mitoplasts from ρ0 mitochondria display PTP currents indistinguishable from their wild-type counterparts. In this work, we show that peripheral stalk subunits are essential to turn the F-ATP synthase into the PTP and that the ANT provides mitochondria with a distinct permeability pathway.

MODES OF TRANSILLUMINATION OF THE DENSE PLASMA LAYER VIA ELECTROMAGNETIC BEAM

Interaction of the powerful electromagnetic ray of the limited radius with a dense plasma layer was studied viacomputer simulation using the PIConGPU software package in 3D geometry. The characteristic modes of channel formation in the barrier (namely laminar and turbulent) are described. Turbulent mode can be associated with the fast transillumination, observed in laboratory experiments. The channel formation time as a function of the incident beam power and the plasma layer density is studied. Information transparency is also observed at the third harmonic of the incident wave.

Intracellular Ion Changes Induced by the Exposure to Beta-Amyloid Can Be Explained by the Formation of Channels in the Lysosomal Membranes

In this manuscript, we reassess the data on beta-amyloid-induced changes of intracellular ions concentrations published previously by Abramov et al. (2003, 2004). Their observations made using high-resolution confocal microscopy with fast temporal resolution of images formed by fluorescent ion-sensitive fluorescent probes in living cells represent an unequivocal support for the amyloid channel theory. However, closer look reveals multiple facts which cannot be explained by channel formation in plasma membrane. Recently proposed amyloid degradation toxicity hypothesis provides the interpretation to these facts by considering that channels are formed in the lysosomal membranes.

Lower Cambrian (Series 2) small shelly fossils from along Nares Strait (Nunavut and Greenland; Laurentia)

Three small assemblages of lower Cambrian (Cambrian Series 2, Stage 4) small shelly fossils are described from Laurentian strata astride Nares Strait. The fauna from the Humboldt Formation of Daugaard-Jensen Land, North Greenland, is derived from inner shelf sediments deposited on the stable craton of the Inglefield Land High. Fossils from Judge Daly Promontory, eastern Ellesmere Island, Nunavut, occur in strata of the Cambrian Ellesmere Group (Kane Basin Formation) that have been structurally juxtaposed against older strata; they were originally assigned to the Kennedy Channel Formation, which is now considered to be of Neoproterozoic age. A similar fauna from offshore environments of the Aftenstjernesø Formation in northern Nyeboe Land, North Greenland, reflects the regional structural and sedimentological continuity with the Canadian Cambrian succession. Pojetaia robsonae sp. nov. is described from Judge Daly Promontory.