The effects of the ω-6 polyunsaturated fatty acid dihomo-γ-linolenic acid on platelet function

PhD ThesisEicosanoids play an essential role in platelet function as they can both stimulate and inhibit platelet reactivity. Current anti-thrombotic therapies include aspirin that inhibits the enzyme COX-1 and thereby the production of arachidonic acid (AA)-derived, pro-aggregatory eicosanoids from platelets. Despite generally promising clinical outcomes, aspirin has limitations in efficacy and is associated with various side effects, highlighting the need for improved therapies. Eicosanoid synthesis can be altered by changes in substrate availability as well as by inhibition of pathway enzymes. For example, the omega-6 fatty acid dihomo-γ-linolenic acid (DGLA) can be utilized by platelets for the synthesis of anti-aggregatory eicosanoids, especially via pathways independent of COX-1, and can thus promote platelet inhibition. The objectives of this thesis were therefore to investigate the effects of DGLA on platelet reactivity and further examine the DGLA effects together with aspirin. In this study, I found that DGLA inhibits a variety of platelet functions, including platelet aggregation, adhesion, spreading and the surface expression of platelet activation markers. I further showed that DGLA inhibits α- and dense granule release and alters the platelet proteomic releasate profile, especially in the presence of aspirin. Pathway enrichment analysis predicted that addition of DGLA would further inhibit platelet-specific responses compared to aspirin alone, and might also influence inflammation and tissue repair, suggesting an exciting potential of DGLA to enhance therapeutic strategies for vascular diseases. Additionally, DGLA caused loss 8 in proteins linked to cytoskeletal rearrangement and initiated a unique physical phenotype in activated platelets similar to unstimulated platelets with curved elongations. Investigating the eicosanoid profile in response to DGLA, I found that DGLA did not alter the production of AA-derived eicosanoids, however, promoted the production of anti-aggregatory series-1 prostaglandins, 12-HETrE and the anti-inflammatory 15-HETrE and 8-HETrE. While aspirin inhibited AA-derived prostanoid production, it only minimally reduced DGLA-derived prostaglandin production, suggesting the involvement of still uncharacterised production pathways

A corrosion control manual for rail rapid transit

In 1979, during the planning stage of the Metropolitan Dade County Transit System, the need was expressed for a corrosion control manual oriented to urban rapid transit system use. This manual responds to that need. The objective of the manual is to aid rail rapid transit agencies by providing practical solutions to selected corrosion problems. The scope of the manual encompasses corrosion problems of the facilities of rapid transit systems: structures and tracks, platforms and stations, power and signals, and cars. It also discusses stray electric current corrosion. Both design and maintenance solutions are provided for each problem. Also included are descriptions of the types of corrosion and their causes, descriptions of rapid transit properties, a list of corrosion control committees and NASA, DOD, and ASTM specifications and design criteria to which reference is made in the manual. A bibliography of papers and excerpts of reports and a glossary of frequency used terms are provided

A Corrosion Control Manual for Rail Rapid Transit

This manual addresses corrosion problems in the design, contruction, and maintenance of rapid transit systems. Design and maintenance solutions are provided for each problem covered. The scope encompasses all facilities of urban rapid transit systems: structures and tracks, platforms and stations, power and signals, and cars. The types of corrosion and their causes as well as rapid transit properties are described. Corrosion control committees, and NASA, DOD, and ASTM specifications and design criteria to which reference is made in the manual are listed. A bibliography of papers and excerpts of reports is provided and a glossary of frequently used terms is included

BETASCAN: Probable β\beta-amyloids Identified by Pairwise Probabilistic Analysis

Amyloids and prion proteins are clinically and biologically important $\beta$-structures, whose supersecondary structures are difficult to determine by standard experimental or computational means. In addition, significant conformational heterogeneity is known or suspected to exist in many amyloid fibrils. Recent work has indicated the utility of pairwise probabilistic statistics in $\beta$-structure prediction. We develop here a new strategy for $\beta$-structure prediction, emphasizing the determination of $\beta$-strands and pairs of $\beta$-strands as fundamental units of $\beta$-structure. Our program, BETASCAN, calculates likelihood scores for potential $\beta$-strands and strand-pairs based on correlations observed in parallel $\beta$-sheets. The program then determines the strands and pairs with the greatest local likelihood for all of the sequence's potential $\beta$-structures. BETASCAN suggests multiple alternate folding patterns and assigns relative a priori probabilities based solely on amino acid sequence, probability tables, and pre-chosen parameters. The algorithm compares favorably with the results of previous algorithms (BETAPRO, PASTA, SALSA, TANGO, and Zyggregator) in $\beta$-structure prediction and amyloid propensity prediction. Accurate prediction is demonstrated for experimentally determined amyloid $\beta$-structures, for a set of known $\beta$-aggregates, and for the parallel $\beta$-strands of $\beta$-helices, amyloid-like globular proteins. BETASCAN is able both to detect $\beta$-strands with higher sensitivity and to detect the edges of $\beta$-strands in a richly $\beta$-like sequence. For two proteins (A$\beta$ and Het-s), there exist multiple sets of experimental data implying contradictory structures; BETASCAN is able to detect each competing structure as a potential structure variant. The ability to correlate multiple alternate $\beta$-structures to experiment opens the possibility of computational investigation of prion strains and structural heterogeneity of amyloid. BETASCAN is publicly accessible on the Web at http://betascan.csail.mit.edu

Conservation of the PBL-RBOH immune module in land plants

The rapid production of reactive oxygen species (ROS) is a key signaling output in plant immunity. In the angiosperm model species Arabidopsis thaliana (hereafter Arabidopsis), recognition of non- or altered-self elicitor patterns by cell-surface immune receptors activates the receptor-like cytoplasmic kinases (RLCKs) of the AVRPPHB SUSCEPTIBLE 1 (PBS1)-like (PBL) family, particularly BOTRYTIS-INDUCED KINASE1 (BIK1).$^{1}$$^{,}$$^{2}$$^{,}$$^{3}$ BIK1/PBLs in turn phosphorylate the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase RESPIRATORY BURST OXIDASE HOMOLOG D (RBOHD) to induce apoplastic ROS production.$^{4}$$^{,}$$^{5}$ PBL and RBOH functions in plant immunity have been extensively characterized in flowering plants. Much less is known about the conservation of pattern-triggered ROS signaling pathways in non-flowering plants. In this study, we show that in the liverwort Marchantia polymorpha (hereafter Marchantia), single members of the RBOH and PBL families, namely MpRBOH1 and MpPBLa, are required for chitin-induced ROS production. MpPBLa directly interacts with and phosphorylates MpRBOH1 at specific, conserved sites within its cytosolic N terminus, and this phosphorylation is essential for chitin-induced MpRBOH1-mediated ROS production. Collectively, our work reveals the functional conservation of the PBL-RBOH module that controls pattern-triggered ROS production in land plants

Is It Easy to Be Urban? Convergent Success in Urban Habitats among Lineages of a Widespread Native Ant

The most rapidly expanding habitat globally is the urban habitat, yet the origin and life histories of the populations of native species that inhabit this habitat remain poorly understood. We use DNA barcoding of the COI gene in the widespread native pest ant Tapinoma sessile to test two hypotheses regarding the origin of urban populations and traits associated with their success. First, we determine if urban samples of T. sessile have a single origin from natural populations by looking at patterns of haplotype clustering from across their range. Second, we examine whether polygynous colony structure – a trait associated with invasion success – is correlated with urban environments, by studying the lineage dependence of colony structure. Our phylogenetic analysis of 49 samples identified four well supported geographic clades. Within clades, Kimura-2 parameter pairwise genetic distances revealed <2.3% variation; however, between clade genetic distances were 7.5–10.0%, suggesting the possibility of the presence of cryptic species. Our results indicate that T. sessile has successfully colonized urban environments multiple times. Additionally, polygynous colony structure is a highly plastic trait across habitat, clade, and haplotype. In short, T. sessile has colonized urban habitats repeatedly and appears to do so using life history strategies already present in more natural populations. Whether similar results hold for other species found in urban habitats has scarcely begun to be considered

The phagocytosis oxidase/Bem1p domain-containing protein PB1CP negatively regulates the NADPH oxidase RBOHD in plant immunity

Perception of pathogen-associated molecular patterns (PAMPs) by surface-localized pattern recognition receptors activates RESPIRATORY BURST OXIDASE HOMOLOG D (RBOHD) through direct phosphorylation by BOTRYTIS-INDUCED KINASE 1 (BIK1) and induces the production of reactive oxygen species (ROS). RBOHD activity must be tightly controlled to avoid the detrimental effects of ROS, but little is known about RBOHD downregulation. To understand the regulation of RBOHD, we used co-immunoprecipitation of RBOHD with mass spectrometry analysis and identified PHAGOCYTOSIS OXIDASE/BEM1P (PB1) DOMAIN-CONTAINING PROTEIN (PB1CP). PB1CP negatively regulates RBOHD and the resistance against the fungal pathogen Colletotrichum higginsianum. PB1CP competes with BIK1 for binding to RBOHD in vitro. Furthermore, PAMP treatment enhances the PB1CP-RBOHD interaction, thereby leading to the dissociation of phosphorylated BIK1 from RBOHD in vivo. PB1CP localizes at the cell periphery and PAMP treatment induces relocalization of PB1CP and RBOHD to the same small endomembrane compartments. Additionally, overexpression of PB1CP in Arabidopsis leads to a reduction in the abundance of RBOHD protein, suggesting the possible involvement of PB1CP in RBOHD endocytosis. We found PB1CP, a novel negative regulator of RBOHD, and revealed its possible regulatory mechanisms involving the removal of phosphorylated BIK1 from RBOHD and the promotion of RBOHD endocytosis

Structural Characterization of a High-Temperature, Ionic Conducting Ceramic using Perturbed Angular Correlation Spectroscopy

Perturbed angular correlation (PAC) spectroscopy has been used to characterize several structural aspects of a high-temperature, ionic conducting ceramic, CaZr3.95Hf0.05P6O24. Hafnium was introduced into the material to provide the PAC probe nuclei, 181Hf/181Ta, which were located primarily at Zr sites. PAC measurements were made over a range of temperatures from 77 to 1180 K, and they have been analyzed and interpreted using several simple models. The distorted octahedral crystal field at the Zr site produced a (low-frequency) static electric quadrupole interaction which can be accurately described by the point-charge model. But, the temperature dependence of the associated electric field gradient (EFG) cannot be described accurately by purely static considerations via the point-charge model and high-temperature x-ray diffraction data. Although a high-frequency static interaction was also observed, the measurements were not sufficiently accurate to identify its origin unambiguously. Some of the high-temperature measurements show evidence of a time-varying interaction, which may result from Ca2+-ion jumping. But, jump frequencies derived classically from high-temperature electrical dc conductivity measurements are too low to agree with those indicated by the PAC data. However, the dc conductivity measurements support a simple model of thermally activated Ca2+-ion transport. The temperature dependence of the EFG (corresponding to the low-frequency interaction) was used to determine an effective Debye-Waller factor. As a result of using this approach to analyze this type of PAC data, this factor was shown also to agree qualitatively with the predictions of the Debye crystal model, although significant theoretical limitations were encountered. These particular results suggest that the PAC technique may provide new insights into understanding advanced ceramic materials

Investigating the Stratigraphy of an Alluvial Aquifer Using Crosswell Seismic Traveltime Tomography

In this study, we investigate the use of crosswell P-wave seismic tomography to obtain spatially extensive information about subsurface sedimentary architecture and heterogeneity in alluvial aquifers. Our field site was a research wellfield in an unconfined aquifer near Boise, Idaho. The aquifer consists of a ~ 20-m-thick sequence of alluvial cobble- and-sand deposits, which have been subdivided into five stratigraphic units based on neutron porosity logs, grainsize analysis, and radar reflection data. We collected crosswell and borehole-to-surface seismic data in wells 17.1 m apart. We carefully considered the impact of well deviation, data quality control, and the choice of inversion parameters. Our linearized inverse routine had a curved-ray forward model and used different grids for forward modeling and inversion. An analysis of the model covariance and resolution matrices showed that the velocity models had an uncertainty of \u3c10 m\u3e/s, a vertical resolution of ~ 1 m, and a horizontal resolution of ~ 5 m. The velocity in the saturated zone varied between 2100 m/s and 2700 m/s. Inclusion of the borehole-to-surface data eliminated the Xshaped pattern that is a common artifact in crosswell tomography, and the increased angular coverage also improved the accuracy of the model near the top of the tomogram. The final velocity model is consistent with previous stratigraphic analyses of the site, although the locations of some of the unit boundaries differ by as much as 2 m in places. The results of this study demonstrate that seismic tomography can be used to image the sedimentary architecture of unconsolidated alluvial aquifers, even when the lithologic contrasts between units are subtle