The Role of Alginate in the Inhibition of Macrophage Phagocytosis of Mucoid Pseudomonas aeruginosa

During colonization of the cystic fibrosis airway Pseudomonas aeruginosa converts from non-mucoid to a mucoid phenotype, characterized by the production of the exopolysaccharide alginate. Alginate production has been shown to enhance survival by promoting biofilm formation, evading complement killing, and resisting phagocytosis. The mechanism by which alginate protects P. aeruginosa from phagocytosis is unclear. To investigate the role of alginate in the inhibition of phagocytosis, a human monocytic cell line (THP-1) and a murine alveolar macrophage cell line (MH-S) were used to determine the effects of alginate on macrophage binding, signaling, and phagocytosis. Phagocytosis assays using the mucoid cystic fibrosis clinical isolate FRD1, and its non-mucoid isogenic algD mutant FRD1131, revealed that alginate inhibits opsonic and non-opsonic phagocytosis. The inhibitory effect of alginate production is intrinsic to the bacteria as exogenous alginate was unable to protect non-mucoid FRD1131 from phagocytosis. Decreased binding of FRD1 compared to FRD1131 was also demonstrated by using the actin polymerization inhibitor cytochalasin D to inhibit phagocytosis. Furthermore, studies using blocking antibodies to CD11b and CD14 found that both of these receptors were important for the phagocytosis of FRD, and it is likely that these receptors are blocked by alginate. Alginate production by P. aeruginosa may reduce lipid raft formation, however, it was not found to affect acid sphingomyelinase activity, which is important for ceramide formation within the lipid raft. Decreased binding led to decreased signaling in macrophages demonstrated by reduction in level and alteration in kinetics of phosphorylation of AKT and ERK1/2 kinases. Signaling pathway inhibitors revealed that PI3K, but not MEK, activation was critical for phagocytosis of P. aeruginosa. Despite altered intracellular signaling in murine macrophages, both mucoid and non-mucoid P. aeruginosa induced similar levels of IL-8 and MIP-2 from human and murine macrophages, respectively. By understanding the pathways involved in mediating efficient phagocytosis of clinical isolates, it may be possible to develop a treatment to promote clearance by the resident alveolar macrophages. These experiments may serve as a model to evaluate the effectiveness of such treatments. This approach also provides valuable insight into previously unknown mechanisms of phagocytosis of P. aeruginosa

Solution structure of a bacterial microcompartment targeting peptide and its application in the construction of an ethanol bioreactor

Targeting of proteins to bacterial microcompartments (BMCs) is mediated by an 18-amino-acid peptide sequence. Herein, we report the solution structure of the N-terminal targeting peptide (P18) of PduP, the aldehyde dehydrogenase associated with the 1,2-propanediol utilization metabolosome from Citrobacter freundii. The solution structure reveals the peptide to have a well-defined helical conformation along its whole length. Saturation transfer difference and transferred NOE NMR has highlighted the observed interaction surface on the peptide with its main interacting shell protein, PduK. By tagging both a pyruvate decarboxylase and an alcohol dehydrogenase with targeting peptides, it has been possible to direct these enzymes to empty BMCs in vivo and to generate an ethanol bioreactor. Not only are the purified, redesigned BMCs able to transform pyruvate into ethanol efficiently, but the strains containing the modified BMCs produce elevated levels of alcohol

Reviewing the International Year of Deserts and Desertification 2006: What contribution towards combating global desertification and implementing the united nations convention to combat desertification?

During the United Nations General Assembly's 58th Ordinary Session in 2003, a decision was adopted declaring 2006 the International Year of Deserts and Desertification (IYDD). This paper critically reviews this International Year. It draws on the key outputs from IYDD events from across the globe to highlight the challenges and ways forward in both combating desertification and implementing the United Nations Convention to Combat Desertification (UNCCD). The paper considers what the IYDD outputs mean for the current and historical controversies surrounding the desertification issue and presents an overall evaluation of the successes of IYDD for the different stakeholder groups within the desertification regime. It is concluded that while the International Year can be considered to have met the United Nations's four objectives: to address the long-term oriented implementation of the UNCCD; raise awareness of the implications of desertification; facilitate networking with all stakeholders; and disseminate information relating to the UNCCD, the real challenge lies in moving the IYDD outcomes away from the conferences, meetings and networks that contributed to their generation, towards a more concrete, tangible effort to conserve deserts and effectively monitor and control desertification and land degradation on the ground

Two New Tidally Distorted White Dwarfs

We identify two new tidally distorted white dwarfs (WDs), SDSS J174140.49+652638.7 and J211921.96-001825.8 (hereafter J1741 and J2119). Both stars are extremely low mass (ELM, < 0.2 Msun) WDs in short-period, detached binary systems. High-speed photometric observations obtained at the McDonald Observatory reveal ellipsoidal variations and Doppler beaming in both systems; J1741, with a minimum companion mass of 1.1 Msun, has one of the strongest Doppler beaming signals ever observed in a binary system (0.59 \pm 0.06% amplitude). We use the observed ellipsoidal variations to constrain the radius of each WD. For J1741, the star's radius must exceed 0.074 Rsun. For J2119, the radius exceeds 0.10 Rsun. These indirect radius measurements are comparable to the radius measurements for the bloated WD companions to A-stars found by the Kepler spacecraft, and they constitute some of the largest radii inferred for any WD. Surprisingly, J1741 also appears to show a 0.23 \pm 0.06% reflection effect, and we discuss possible sources for this excess heating. Both J1741 and J2119 are strong gravitational wave sources, and the time-of-minimum of the ellipsoidal variations can be used to detect the orbital period decay. This may be possible on a timescale of a decade or less.Comment: 6 pages, 4 figures, accepted for publication in the Astrophysical Journa

Treatment of Triple-Negative Breast Cancer Cells with the Canady Cold Plasma Conversion System: Preliminary Results

Triple-negative breast cancer is a phenotype of breast cancer where the expression level of estrogen, progesterone and human epidermal growth factor receptor 2 (HER2) receptors are low or absent. It is more frequently diagnosed in younger and premenopausal women, among which African and Hispanic have a higher rate. Cold atmospheric plasma has revealed its promising ant-cancer capacity over the past two decades. In this study, we report the first cold plasma jet delivered by the Canady Cold Plasma Conversion Unit and characterization of its electric and thermal parameters. The unit effectively reduced the viability of triple-negative breast cancer up to 80% without thermal damage, providing a starting point for future clinical trials

FAD binding, cobinamide binding and active site communication in the corrin reductase (CobR)

Adenosylcobalamin, the coenzyme form of vitamin B12, is one Nature's most complex coenzyme whose de novo biogenesis proceeds along either an anaerobic or aerobic metabolic pathway. The aerobic synthesis involves reduction of the centrally chelated cobalt metal ion of the corrin ring from Co(II) to Co(I) before adenosylation can take place. A corrin reductase (CobR) enzyme has been identified as the likely agent to catalyse this reduction of the metal ion. Herein, we reveal how Brucella melitensis CobR binds its coenzyme FAD (flavin dinucleotide) and we also show that the enzyme can bind a corrin substrate consistent with its role in reduction of the cobalt of the corrin ring. Stopped-flow kinetics and EPR reveal a mechanistic asymmetry in CobR dimer that provides a potential link between the two electron reduction by NADH to the single electron reduction of Co(II) to Co(I)

A scalable analytical framework for spatio-temporal analysis of neighborhood change: A sequence analysis approach

© Springer Nature Switzerland AG 2020. Spatio-temporal changes reflect the complexity and evolution of demographic and socio-economic processes. Changes in the spatial distribution of population and consumer demand at urban and rural areas are expected to trigger changes in future housing and infrastructure needs. This paper presents a scalable analytical framework for understanding spatio-temporal population change, using a sequence analysis approach. This paper uses gridded cell Census data for Great Britain from 1971 to 2011 with 10-year intervals, creating neighborhood typologies for each Census year. These typologies are then used to analyze transitions of grid cells between different types of neighborhoods and define representative trajectories of neighborhood change. The results reveal seven prevalent trajectories of neighborhood change across Great Britain, identifying neighborhoods which have experienced stable, upward and downward pathways through the national socioeconomic hierarchy over the last four decades

ADAM10 is essential for Notch2-dependent marginal zone B cell development and CD23 cleavage in vivo

The proteolytic activity of a disintegrin and metalloproteinase 10 (ADAM10) regulates cell-fate decisions in Drosophila and mouse embryos. However, in utero lethality of ADAM10−/− mice has prevented examination of ADAM10 cleavage events in lymphocytes. To investigate their role in B cell development, we generated B cell–specific ADAM10 knockout mice. Intriguingly, deletion of ADAM10 prevented development of the entire marginal zone B cell (MZB) lineage. Additionally, cleavage of the low affinity IgE receptor, CD23, was profoundly impaired, but subsequent experiments demonstrated that ADAM10 regulates CD23 cleavage and MZB development by independent mechanisms. Development of MZBs is dependent on Notch2 signaling, which requires proteolysis of the Notch2 receptor by a previously unidentified proteinase. Further experiments revealed that Notch2 signaling is severely impaired in ADAM10-null B cells. Thus, ADAM10 critically regulates MZB development by initiating Notch2 signaling. This study identifies ADAM10 as the in vivo CD23 sheddase and an important regulator of B cell development. Moreover, it has important implications for the treatment of numerous CD23- and Notch-mediated pathologies, ranging from allergy to cancer

An Ordered Inheritance Strategy for the Golgi Apparatus: Visualization of Mitotic Disassembly Reveals a Role for the Mitotic Spindle

During mitosis, the ribbon of the Golgi apparatus is transformed into dispersed tubulo-vesicular membranes, proposed to facilitate stochastic inheritance of this low copy number organelle at cytokinesis. Here, we have analyzed the mitotic disassembly of the Golgi apparatus in living cells and provide evidence that inheritance is accomplished through an ordered partitioning mechanism. Using a Sar1p dominant inhibitor of cargo exit from the endoplasmic reticulum (ER), we found that the disassembly of the Golgi observed during mitosis or microtubule disruption did not appear to involve retrograde transport of Golgi residents to the ER and subsequent reorganization of Golgi membrane fragments at ER exit sites, as has been suggested. Instead, direct visualization of a green fluorescent protein (GFP)-tagged Golgi resident through mitosis showed that the Golgi ribbon slowly reorganized into 1–3-μm fragments during G2/early prophase. A second stage of fragmentation occurred coincident with nuclear envelope breakdown and was accompanied by the bulk of mitotic Golgi redistribution. By metaphase, mitotic Golgi dynamics appeared to cease. Surprisingly, the disassembly of mitotic Golgi fragments was not a random event, but involved the reorganization of mitotic Golgi by microtubules, suggesting that analogous to chromosomes, the Golgi apparatus uses the mitotic spindle to ensure more accurate partitioning during cytokinesis