Examining the Effect of Pore Size Distribution and Shape on Flow through Unsaturated Peat using Computer Tomography

The hydraulic conductivity of unsaturated peat soil is controlled by the air-filled porosity, pore size and geometric distribution as well as other physical properties of peat materials. This study investigates how the size and shape of pores affects the flow of water through peat soils. In this study we used X-ray Computed Tomography (CT), at 45μm resolution under 5 specific soil-water pressure head levels to provide 3-D, high-resolution images that were used to detect the inner pore structure of peat samples under a changing water regime. Pore structure and configuration were found to be irregular, which affected the rate of water transmission through peat soils. The 3-D analysis suggested that pore distribution is dominated by a single large pore-space. At low pressure head, this single large air-filled pore imparted a more effective flowpath compared to smaller pores. Smaller pores were disconnected and the flowpath was more tortuous than in the single large air-filled pore, and their contribution to flow was negligible when the single large pore was active. We quantify the pore structure of peat soil that affects the hydraulic conductivity in the unsaturated condition, and demonstrate the validity of our estimation of peat unsaturated hydraulic conductivity by making a comparison with a standard permeameter-based method. Estimates of unsaturated hydraulic conductivities were made for the purpose of testing the sensitivity of pore shape and geometry parameters on the hydraulic properties of peats and how to evaluate the structure of the peat and its affects on parameterization. We also studied the ability to quantify these factors for different soil moisture contents in order to define how the factors controlling the shape coefficient vary with changes in soil water pressure head. The relation between measured and estimated unsaturated hydraulic conductivity at various heads shows that rapid initial drainage, that changes the air-filled pore properties, creates a sharp decline in hydraulic conductivity. This is because the large pores readily lose water, the peat rapidly becomes less conductive and the flow path among pores, more tortuous

Peptide Orientation Affects Selectivity in Ion-Exchange Chromatography

Here we demonstrate that separation of proteolytic peptides, having the same net charge and one basic residue, is affected by their specific orientation toward the stationary phase in ion-exchange chromatography. In electrostatic repulsion−hydrophilic interaction chromatography (ERLIC) with an anion-exchange material, the C-terminus of the peptides is, on average, oriented toward the stationary phase. In cation exchange, the average peptide orientation is the opposite. Data with synthetic peptides, serving as orientation probes, indicate that in tryptic/Lys-C peptides the C-terminal carboxyl group appears to be in a zwitterionic bond with the side chain of the C-terminal Lys/Arg residue. In effect, the side chain is then less basic than the N-terminus, accounting for the specific orientation of tryptic and Lys-C peptides. Analyses of larger sets of peptides, generated from lysates by either Lys-N, Lys-C, or trypsin, reveal that specific peptide orientation affects the ability of charged side chains, such as phosphate residues, to influence retention. Phosphorylated residues that are remote in the sequence from the binding site affect retention less than those that are closer. When a peptide contains multiple charged sites, then orientation is observed to be less rigid and retention tends to be governed by the peptide’s net charge rather than its sequence. These general observations could be of value in confirming a peptide’s identification and, in particular, phosphosite assignments in proteomics analyses. More generally, orientation accounts for the ability of chromatography to separate peptides of the same composition but different sequence

Analysis of hybrid mode-locking of two-section quantum dot lasers operating at 1.5 micron

For the first time a detailed study of hybrid mode-locking in two- section InAs/InP quantum dot Fabry-Pérot-type lasers is presented. The output pulses have a typical upchirp of approximately 8 ps/nm, leading to very elongated pulses. The mechanism leading to this typical pulse shape and the phase noise is investigated by detailed radio-frequency and optical spectral studies as well as time-domain studies. The pulse shaping mechanism in these lasers is found to be fundamentally different than the mechanism observed in conventional mode-locked laser diodes, based on quantum well gain or bulk material. ©2009 Optical Society of America

Peeling the Otolith of Fish: Optimal Parameterization for Micro-CT Scanning

In this paper, we aim to provide optimal parameters for micro-computed tomography scans of fish otoliths. We tested fifteen different combinations to sagittae. The images were scaled to Hounsfield units, and segmented in two distinct volumes-of-interest (external and internal). The strategy we applied, for identifying optimum scan settings for otoliths, included analyses of the sinogram, the distribution of the Hounsfield units and the signal-to-noise ratio. Based on these tests, the optimum sets of parameters for the acquisition of tomographic images of sagittal otoilths were 80 kV, 220 μA, and 0.5 mm aluminum filter. The method allowed 3D shape analysis, internal and external density distribution, layer-by-layer density segmentation, and a potential objective method to count growth rings in otoliths. It was possible to compare mean densities between species, and we observed a significant difference among them. In addition, there are ontogenic changes, which could be increasing or decreasing the density. In this study, we applied tomography for several otolith analysis, that could be of great interest for future studies in diverse areas that use otoliths as the basic structure of analysis, or represents a new research line called eco-densitometry of otoliths, where tomography could be applied to explore the density within an ecological perspective.Fil: Vasconcelos Filho, Jonas. Universidad Federal Rural Pernambuco; BrasilFil: Thomsen, Felix Sebastian Leo. Universidad Nacional del Sur. Departamento de Ingeniería Eléctrica y de Computadoras; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; ArgentinaFil: Stosic, Borko. Universidad Federal Rural Pernambuco; BrasilFil: Antonino, Antonio C. D.. Universidad Federal Rural Pernambuco; BrasilFil: Duarte, Daniel A.. Universidad Federal Rural Pernambuco; BrasilFil: Heck, Richard J.. University of Guelph; CanadáFil: Lessa, Rosangela P. T.. Universidad Federal Rural Pernambuco; BrasilFil: Santana, Francisco M.. Universidad Federal Rural Pernambuco; BrasilFil: Ferreira, Beatrice P.. Universidad Federal Rural Pernambuco; BrasilFil: Duarte Neto, Paulo J.. Universidad Federal Rural Pernambuco; Brasi

Human marginal zone B cell development from early T2 progenitors.

B cells emerge from the bone marrow as transitional (TS) B cells that differentiate through T1, T2, and T3 stages to become naive B cells. We have identified a bifurcation of human B cell maturation from the T1 stage forming IgMhi and IgMlo developmental trajectories. IgMhi T2 cells have higher expression of α4β7 integrin and lower expression of IL-4 receptor (IL4R) compared with the IgMlo branch and are selectively recruited into gut-associated lymphoid tissue. IgMhi T2 cells also share transcriptomic features with marginal zone B cells (MZBs). Lineage progression from T1 cells to MZBs via an IgMhi trajectory is identified by pseudotime analysis of scRNA-sequencing data. Reduced frequency of IgMhi gut-homing T2 cells is observed in severe SLE and is associated with reduction of MZBs and their putative IgMhi precursors. The collapse of the gut-associated MZB maturational axis in severe SLE affirms its existence in health

Oxonium Ion-Guided Optimization of Ion Mobility-Assisted Glycoproteomics on the timsTOF Pro

Spatial separation of ions in the gas phase, providing information about their size as collisional cross-sections, can readily be achieved through ion mobility. The timsTOF Pro (Bruker Daltonics) series combines a trapped ion mobility device with a quadrupole, collision cell, and a time-of-flight analyzer to enable the analysis of ions at great speed. Here, we show that the timsTOF Pro is capable of physically separating N-glycopeptides from nonmodified peptides and producing high-quality fragmentation spectra, both beneficial for glycoproteomics analyses of complex samples. The glycan moieties enlarge the size of glycopeptides compared with nonmodified peptides, yielding a clear cluster in the mobilogram that, next to increased dynamic range from the physical separation of glycopeptides and nonmodified peptides, can be used to make an effective selection filter for directing the mass spectrometer to analytes of interest. We designed an approach where we (1) focused on a region of interest in the ion mobilogram and (2) applied stepped collision energies to obtain informative glycopeptide tandem mass spectra on the timsTOF Pro:glyco-polygon–stepped collision energy-parallel accumulation serial fragmentation. This method was applied to selected glycoproteins, human plasma– and neutrophil-derived glycopeptides. We show that the achieved physical separation in the region of interest allows for improved extraction of information from the samples, even at shorter liquid chromatography gradients of 15 min. We validated our approach on human neutrophil and plasma samples of known makeup, in which we captured the anticipated glycan heterogeneity (paucimannose, phosphomannose, high mannose, hybrid and complex glycans) from plasma and neutrophil samples at the expected abundances. As the method is compatible with off-the-shelve data acquisition routines and data analysis software, it can readily be applied by any laboratory with a timsTOF Pro and is reproducible as demonstrated by a comparison between two laboratories

The PRoteomics IDEntification (PRIDE) Converter 2 Framework: An Improved Suite of Tools to Facilitate Data Submission to the PRIDE Database and the ProteomeXchange Consortium

The original PRIDE Converter tool greatly simplified the process of submitting mass spectrometry (MS)-based proteomics data to the PRIDE database. However, after much user feedback, it was noted that the tool had some limitations and could not handle several user requirements that were now becoming commonplace. This prompted us to design and implement a whole new suite of tools that would build on the successes of the original PRIDE Converter and allow users to generate submission-ready, well-annotated PRIDE XML files. The PRIDE Converter 2 tool suite allows users to convert search result files into PRIDE XML (the format needed for performing submissions to the PRIDE database), generate mzTab skeleton files that can be used as a basis to submit quantitative and gel-based MS data, and post-process PRIDE XML files by filtering out contaminants and empty spectra, or by merging several PRIDE XML files together. All the tools have both a graphical user interface that provides a dialog-based, user-friendly way to convert and prepare files for submission, as well as a command-line interface that can be used to integrate the tools into existing or novel pipelines, for batch processing and power users. The PRIDE Converter 2 tool suite will thus become a cornerstone in the submission process to PRIDE and, by extension, to the ProteomeXchange consortium of MS-proteomics data repositories.publishedVersio

Spatial differentiation of gene expression in Aspergillus niger colony grown for sugar beet pulp utilization

Citation: Benoit, I., Zhou, M. M., Duarte, A. V., Downes, D. J., Todd, R. B., Kloezen, W., . . . de Vries, R. P. (2015). Spatial differentiation of gene expression in Aspergillus niger colony grown for sugar beet pulp utilization. Scientific Reports, 5(1), 1-13. https://doi.org/10.1038/srep13592Degradation of plant biomass to fermentable sugars is of critical importance for the use of plant materials for biofuels. Filamentous fungi are ubiquitous organisms and major plant biomass degraders. Single colonies of some fungal species can colonize massive areas as large as five soccer stadia. During growth, the mycelium encounters heterogeneous carbon sources. Here we assessed whether substrate heterogeneity is a major determinant of spatial gene expression in colonies of Aspergillus niger. We analyzed whole-genome gene expression in five concentric zones of 5-day-old colonies utilizing sugar beet pulp as a complex carbon source. Growth, protein production and secretion occurred throughout the colony. Genes involved in carbon catabolism were expressed uniformly from the centre to the periphery whereas genes encoding plant biomass degrading enzymes and nitrate utilization were expressed differentially across the colony. A combined adaptive response of carbon-catabolism and enzyme production to locally available monosaccharides was observed. Finally, our results demonstrate that A. niger employs different enzymatic tools to adapt its metabolism as it colonizes complex environments