Material properties of interfacial films of mucoid and nonmucoid Pseudomonas aeruginosa isolates

An understanding of the material properties of biofilms is important when describing how biofilms physically interact with their environment. In this study, aerobic biofilms of Pseudomonas aeruginosa PAO1 and anaerobic sulfate-reducing bacteria (SRB) biofilms of Desulfovibrio sp. EX265 were grown under different fluid shear stresses (τg) in a chemostat recycle loop. Individual biofilm microcolonies were deformed by varying the fluid wall shear stress (τw). The deformation was quantified in terms of strain (ε), and the relative strength of the biofilms was assessed using an apparent elastic coefficient (Eapp) and residual strain (εr) after three cycles of deformation. Aluminium chloride (AlCl3) was then added to both sets of biofilm and the tests repeated. Biofilms grown under higher shear were more rigid and had a greater yield shear stress than those grown under lower shear. The addition of AlCl3 resulted in a significant increase in Eapp and also increased the yield point. We conclude that the strength of the biofilm is in part dependent on the shear under which the biofilm was grown and that the material properties of the biofilm may be manipulated through cation cross-linking of the extracellular polysaccharide (EPS) slime matrix.

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Alginate Polymerization and Modification Are Linked in Pseudomonas aeruginosa

mBio ◽

10.1128/mbio.00453-15 ◽

2015 ◽

Vol 6 (3) ◽

Cited By ~ 33

Author(s):

M. Fata Moradali ◽

Ivan Donati ◽

Ian M. Sims ◽

Shirin Ghods ◽

Bernd H. A. Rehm

Keyword(s):

Pseudomonas Aeruginosa ◽

Material Properties ◽

Molecular Mechanisms ◽

Cell Aggregation ◽

Activation Mechanism ◽

Cellulose Synthesis ◽

Multiprotein Complex ◽

Site Specific ◽

Content Type

ABSTRACTThe molecular mechanisms of alginate polymerization/modification/secretion by a proposed envelope-spanning multiprotein complex are unknown. Here, bacterial two-hybrid assays and pulldown experiments showed that the catalytic subunit Alg8 directly interacts with the proposed copolymerase Alg44 while embedded in the cytoplasmic membrane. Alg44 additionally interacts with the lipoprotein AlgK bridging the periplasmic space. Site-specific mutagenesis of Alg44 showed that protein-protein interactions and stability were independent of conserved amino acid residues R17 and R21, which are involved in c-di-GMP binding, the N-terminal PilZ domain, and the C-terminal 26 amino acids. Site-specific mutagenesis was employed to investigate the c-di-GMP-mediated activation of alginate polymerization by the PilZAlg44domain and Alg8. Activation was found to be different from the proposed activation mechanism for cellulose synthesis. The interactive role of Alg8, Alg44, AlgG (epimerase), and AlgX (acetyltransferase) on alginate polymerization and modification was studied by using site-specific deletion mutants, inactive variants, and overproduction of subunits. The compositions, molecular masses, and material properties of resulting novel alginates were analyzed. The molecular mass was reduced by epimerization, while it was increased by acetylation. Interestingly, when overproduced, Alg44, AlgG, and the nonepimerizing variant AlgG(D324A) increased the degree of acetylation, while epimerization was enhanced by AlgX and its nonacetylating variant AlgX(S269A). Biofilm architecture analysis showed that acetyl groups promoted cell aggregation while nonacetylated polymannuronate alginate promoted stigmergy. Overall, this study sheds new light on the arrangement of the multiprotein complex involved in alginate production. Furthermore, the activation mechanism and the interplay between polymerization and modification of alginate were elucidated.IMPORTANCEThis study provides new insights into the molecular mechanisms of the synthesis of the unique polysaccharide, alginate, which not only is an important virulence factor of the opportunistic human pathogenPseudomonas aeruginosabut also has, due to its material properties, many applications in medicine and industry. Unraveling the assembly and composition of the alginate-synthesizing and envelope-spanning multiprotein complex will be of tremendous significance for the scientific community. We identified a protein-protein interaction network inside the multiprotein complex and studied its relevance with respect to alginate polymerization/modification as well as the c-di-GMP-mediated activation mechanism. A relationship between alginate polymerization and modification was shown. Due to the role of alginate in pathogenesis as well as its unique material properties harnessed in numerous applications, results obtained in this study will aid the design and development of inhibitory drugs as well as the commercial bacterial production of tailor-made alginates.

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TEM studies of amorphization processes and amorphous structures

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100104303 ◽

1988 ◽

Vol 46 ◽

pp. 448-449

Author(s):

T. E. Mitchell ◽

R. B. Schwarz

Keyword(s):

Amorphous Materials ◽

Fission Products ◽

Rapid Quenching ◽

Surface Films ◽

List Type ◽

Oxide Glasses ◽

Crystalline Materials ◽

Amorphous Structures ◽

Amorphous Surface ◽

Interfacial Films

Traditional oxide glasses occur naturally as obsidian and can be made easily by suitable cooling histories. In the past 30 years, a variety of techniques have been discovered which amorphize normally crystalline materials such as metals. These include [1-3]:Rapid quenching from the vapor phase.Rapid quenching from the liquid phase.Electrodeposition of certain alloys, e.g. Fe-P.Oxidation of crystals to produce amorphous surface oxide layers.Interdiffusion of two pure crystalline metals.Hydrogen-induced vitrification of an intermetal1ic.Mechanical alloying and ball-milling of intermetal lie compounds.Irradiation processes of all kinds using ions, electrons, neutrons, and fission products.We offer here some general comments on the use of TEM to study these materials and give some particular examples of such studies.Thin specimens can be prepared from bulk homogeneous materials in the usual way. Most often, however, amorphous materials are in the form of surface films or interfacial films with different chemistry from the substrates.

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Microstructural Evaluation of Silicon Carbide Whisker Reinforced Alumina Fabricated with Carbon-Coated Whiskers

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100088919 ◽

1991 ◽

Vol 49 ◽

pp. 920-921

Author(s):

K. B. Alexander ◽

P. F. Becher

Keyword(s):

Silicon Carbide ◽

High Resolution Electron Microscopy ◽

Ceramic Composites ◽

Interface Debonding ◽

Resolution Electron ◽

Microstructural Evaluation ◽

Carbon Coated ◽

Electron Energy Loss ◽

Interfacial Films ◽

Debonding Process

The presence of interfacial films at the whisker-matrix interface can significantly influence the fracture toughness of ceramic composites. The film may alter the interface debonding process though changes in either the interfacial fracture energy or the residual stress at the interface. In addition, the films may affect the whisker pullout process through the frictional sliding coefficients or the extent of mechanical interlocking of the interface due to the whisker surface topography.Composites containing ACMC silicon carbide whiskers (SiCw) which had been coated with 5-10 nm of carbon and Tokai whiskers coated with 2 nm of carbon have been examined. High resolution electron microscopy (HREM) images of the interface were obtained with a JEOL 4000EX electron microscope. The whisker geometry used for HREM imaging is described in Reference 2. High spatial resolution (< 2-nm-diameter probe) parallel-collection electron energy loss spectroscopy (PEELS) measurements were obtained with a Philips EM400T/FEG microscope equipped with a Gatan Model 666 spectrometer.

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Grain boundary segregation in metals

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100130651 ◽

1992 ◽

Vol 50 (2) ◽

pp. 1204-1205

Author(s):

C.L. Briant

Keyword(s):

Fracture Surface ◽

Grain Boundary ◽

Grain Boundaries ◽

Material Properties ◽

Electron Spectroscopy ◽

High Vacuum ◽

Boundary Segregation ◽

Grain Boundary Segregation ◽

Local Concentration ◽

The One

Grain boundary segregation is the process by which solute elements in a material diffuse to the grain boundaries, become trapped there, and increase their local concentration at the boundary over that in the bulk. As a result of this process this local concentration of the segregant at the grain boundary can be many orders of magnitude greater than the bulk concentration of the segregant. The importance of this problem lies in the fact that grain boundary segregation can affect many material properties such as fracture, corrosion, and grain growth.One of the best ways to study grain boundary segregation is with Auger electron spectroscopy. This spectroscopy is an extremely surface sensitive technique. When it is used to study grain boundary segregation the sample must first be fractured intergranularly in the high vacuum spectrometer. This fracture surface is then the one that is analyzed. The development of scanning Auger spectrometers have allowed researchers to first image the fracture surface that is created and then to perform analyses on individual grain boundaries.

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Relationships Between Grain Boundary Structure and Material Properties

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s1431927600001550 ◽

1980 ◽

Vol 38 ◽

pp. 366-369

Author(s):

Brian Ralph ◽

Barlow Claire ◽

Nicola Ecob

Keyword(s):

Grain Boundary ◽

Material Properties ◽

Main Property ◽

Grain Structure ◽

Boundary Structure ◽

Grain Boundary Structure ◽

Property Changes

This brief review seeks to summarize some of the main property changes which may be induced by altering the grain structure of materials. Where appropriate an interpretation is given of these changes in terms of current theories of grain boundary structure, and some examples from current studies are presented at the end of this paper.

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