Self-assembling multimeric integrin alpha5beta1 ligands for cell attachment and spreading

Substrates utilising clustered arginine-glycine-aspartic acid (RGD) ligand displays support greater cell adhesion over random displays. However, cell adhesion to integrin alpha5beta1 requires the synergy site on the 9th type III fibronectin domain (FIII) in addition to RGD on the 10th FIII domain. Here, we have designed and expressed soluble protein chimeras consisting of an N-terminal 9th-10th FIII domain pair, IgG-derived hinge and leucine zipper-derived helix; the latter mutated to yield di-, tri- and tetrameric coiled coils and thus self-assembling, multimeric integrin alpha5beta1 ligands. A unique C-terminal cysteine was appended to the helix to facilitate 'anchoring' of the chimeras with a defined orientation on a surface. Size-exclusion chromatography and circular dichroism demonstrated that the chimeras self-assembled as multimers in solution with defined secondary structures predicted from theoretical calculations. Biotinylation via a thioether bond was used to selectively bind the chimeras to streptavidin-coated surfaces, each of which was then shown to bind integrin alpha5beta1 by surface plasmon resonance. Spreading of fibroblasts to surfaces derivatised with the chimeras was found to proceed in the order: tetramer > trimer > dimer > monomer. Thus, we describe novel polyvalent integrin alpha5beta1 ligands for facile derivatisation of substrates to improve cell adhesion in vitro.     

Talking with some directors of the U.S. national laboratories

JOM surveyed the leaders of the U.S. Department of Energy’s (DOE) national laboratories for perspectives on some of the U.S. science and engineering community’s most pressing issues regarding R&D, globalization, national security, and budget constraints. The journal posed the same five questions to each lab director. What follows are responses from six of the directors: Michael R. Anastasio, Lawrence Livermore National Laboratory; Dan Arvizu, National Renewable Energy Laboratory (NREL); John J. Grossenbacher, Idaho National Laboratory (INL); Thomas O. Hunter, Sandia National Laboratories; Robert Rosner, Argonne National Laboratory; and Jeffrey Wadsworth, Oak Ridge National Laboratory (ORNL). Kelly Roncone is news editor for JOM.     

Computational modeling of cathodic limitations on localized corrosion of wetted SS 316L at room temperature

The ability of a SS 316L surface wetted with a thin electrolyte layer to serve as an effective cathode for an active localized corrosion site was studied computationally. The dependence of the total net cathodic current, I_net, supplied at the repassivation potential E_rp (of the anodic crevice) on relevant physical parameters including water layer thickness (WL), chloride concentration ([Cl⁻]) and length of cathode (Lc) were investigated using a three-level, full factorial design. The effects of kinetic parameters including the exchange current density (i_o,c) and Tafel slope (β_c) of oxygen reduction, the anodic passive current density (i_p) (on the cathodic surface), and E_rp were studied as well using three-level full factorial designs of [Cl⁻] and Lc with a fixed WL of 25 μm. The study found that all the three parameters WL, [Cl⁻] and Lc as well as the interactions of Lc × WL and Lc × [Cl⁻] had significant impact on I_net. A five-factor regression equation was obtained which fits the computation results reasonably well, but demonstrated that interactions are more complicated than can be explained with a simple linear model. Significant effects on I_net were found upon varying either i_o,c, β_c, or E_rp, whereas i_p in the studied range was found to have little impact. It was observed that I_net asymptotically approached maximum values (I_max) when Lc increased to critical minimum values. I_max can be used to determine the stability of coupled localized corrosion and the critical Lc provides important information for experimental design and corrosion protection.     

Secondary Metabolic Profile as a Tool for Distinction and Characterization of Cultivars of Black Pepper (Piper nigrum L.) Cultivated in Pará State,Brazil

This study evaluated the chemical compositions of the leaves and fruits of eight black pepper cultivars cultivated in Pará State (Amazon, Brazil). Hydrodistillation and gas chromatography–mass spectrometry were employed to extract and analyze the volatile compounds, respectively. Sesquiterpene hydrocarbons were predominant (58.5–90.9%) in the cultivars “Cingapura”, “Equador”, “Guajarina”, “Iaçará”, and “Kottanadan”, and “Bragantina”, “Clonada”, and “Uthirankota” displayed oxygenated sesquiterpenoids (50.6–75.0%). The multivariate statistical analysis applied using volatile composition grouped the samples into four groups: γ-Elemene, curzerene, and δ-elemene (“Equador”/“Guajarina”, I); δ-elemene (“Iaçará”/“Kottanadan”/“Cingapura”, II); elemol (“Clonada”/“Uthirankota”, III) and α-muurolol, bicyclogermacrene, and cubebol (“Bragantina”, IV). The major compounds in all fruit samples were monoterpene hydrocarbons such as α-pinene, β-pinene, and limonene. Among the cultivar leaves, phenolics content (44.75–140.53 mg GAE·g⁻¹ FW), the enzymatic activity of phenylalanine-ammonia lyase (20.19–57.22 µU·mL⁻¹), and carotenoids (0.21–2.31 µg·mL⁻¹) displayed significant variations. Due to black pepper’s susceptibility to Fusarium infection, a molecular docking analysis was carried out on Fusarium protein targets using each cultivar’s volatile components. F. oxysporum endoglucanase was identified as the preferential protein target of the compounds. These results can be used to identify chemical markers related to the susceptibility degree of black pepper cultivars to plant diseases prevalent in Pará State.     

The Innate Immune Glycoprotein Lactoferrin Represses the Helicobacter pylori cag Type IV Secretion System

Chronic infection with Helicobacter pylori increases risk of gastric diseases including gastric cancer. Despite development of a robust immune response, H. pylori persists in the gastric niche. Progression of gastric inflammation to serious disease outcomes is associated with infection with H. pylori strains which encode the cag Type IV Secretion System (cag T4SS). The cag T4SS is responsible for translocating the oncogenic protein CagA into host cells and inducing pro-inflammatory and carcinogenic signaling cascades. Our previous work demonstrated that nutrient iron modulates the activity of the T4SS and biogenesis of T4SS pili. In response to H. pylori infection, the host produces a variety of antimicrobial molecules, including the iron-binding glycoprotein, lactoferrin. Our work shows that apo-lactoferrin exerts antimicrobial activity against H. pylori under iron-limited conditions, while holo-lactoferrin enhances bacterial growth. Culturing H. pylori in the presence of holo-lactoferrin prior to co-culture with gastric epithelial cells, results in repression of the cag T4SS activity. Concomitantly, a decrease in biogenesis of cag T4SS pili at the host-pathogen interface was observed under these culture conditions by high-resolution electron microscopy analyses. Taken together, these results indicate that acquisition of alternate sources of nutrient iron plays a role in regulating the pro-inflammatory activity of a bacterial secretion system and present novel therapeutic targets for the treatment of H. pylori-related disease.     

Model for microbiologically influenced corrosion potential assessment for the oil and gas industry

Corrosion is one of the major causes of failure in onshore and offshore oil and gas operations. Microbiologically influenced corrosion (MIC) is inherently more complex to predict, detect and measure because, for instance, the presence of biofilm and/or bacterial products is not sufficient to indicate active microbiological corrosion. The major challenge for current MIC models is to correlate factors that influence corrosion (i.e. chemical, physical, biological and molecular variables) with the potential of having MIC. Previous work has proposed the potential for MIC as a simple product of multiple factors, without fully considering the synergy or the interference among the factors. The present work proposes a network-based approach to analyse and predict MIC potential considering the complex interactions among a total of 60 influencing factors and 20 screening parameters. The proposed model has the ability to capture the complex interdependences and the synergic interactions of the factors used to assess MIC potential and uses an object-oriented approach based on a Bayesian Network. The model has been tested and verified using real data from a pipeline leakage incident that was a result of MIC. The proposed model constitutes a significant step in deepening the understanding of when MIC occurs and its predictability.

List of acronyms: APB: acid producing bacteria; Aw: water activity; BN: Bayesian network; MIC: microbiologically influenced corrosion; MMMs: molecular microbiological methods; NRB: nitrate-reducing bacteria; OOBN: object-oriented Bayesian network; PWRI: produced water re-injection; SPs: screening parameters; SRB: sulphate-reducing bacteria; SRPs: sulphate-reducing prokaryotes; TDSs: total dissolved solids     

Design and implementation of a continuous microwave heating system for ballast water treatment

A continuous microwave system to treat ballast water inoculated with different invasive species was designed and installed atthe Louisiana State University Agricultural Center. The effectiveness of the system to deliver the required heating loads to inactivate the organisms present was studied. The targeted organisms were microalgae (Nannochloropsis oculata), zooplankton at two different growth stages (newly hatched brine shrimp-Artemia nauplii and adult Artemia), and oyster larvae (Crassosstrea virginica). The system was tested at two different flow rates (1 and 2 liters per min) and power levels (2.5 and 4.5 kW). Temperature profiles indicate that, depending on the species present and the growth stage, the maximum temperature increase will vary from 11.8 to 64.9 degrees C. The continuous microwave heating system delivered uniform and near-instantaneous heating at the outlet proving its effectiveness. The power absorbed and power efficiency varied for the species present. More than 80% power utilization efficiency was obtained at all flow rate and microwave power combinations for microalgae, Artemia nauplii and adults. Test results indicated that microwave treatment can be an effective tool for ballast water treatment, and current high treatment costs notwithstanding, this technique can be added as supplemental technology to the palette of existing treatment methods.     

High resolution spatiotemporal patterns of flow at the landscape scale in montane non-perennial streams

Intermittent and ephemeral streams in dryland environments support diverse assemblages of aquatic and terrestrial life. Understanding when and where water flows provide insights into the availability of water, its response to external controlling factors, and potential sensitivity to climate change and a host of human activities. Knowledge regarding the timing of drying/wetting cycles can also be useful to map critical habitats for species and ecosystems that rely on these temporary water sources. However, identifying the locations and monitoring the timing of streamflow and channel sediment moisture remains a challenging endeavor. In this paper, we analyzed daily conductivity from 37 sensors distributed along 10 streams across an arid mountain front in Arizona (United States) to assess spatiotemporal patterns in flow permanence, defined as the timing and extent of water in streams. Conductivity sensors provide information on surface flow and sediment moisture, supporting a stream classification based on seasonal flow dynamics. Our results provide insight into flow responses to seasonal rainfall, highlighting stream reaches very reactive to rainfall versus those demonstrating more stable streamflow. The strength of stream responses to precipitation are explored in the context of surficial geology. In summary, conductivity data can be used to map potential stream habitat for water-dependent species in both space and time, while also providing the basis upon which sensitivity to ongoing climate change can be evaluated.