High-throughput Analysis of Spatio-temporal multicellular dynamics

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Random walk and cell morphology dynamics in Naegleria gruberi

Amoeboid cell movement and migration are wide-spread across various cell types and species. Microscopy-based analysis of the model systems Dictyostelium and neutrophils over the years have uncovered generality in their overall cell movement pattern. Under no directional cues, the centroid movement can be quantitatively characterized by their persistence to move in a straight line and the frequency of re-orientation. Mathematically, the cells essentially behave as a persistent random walker with memory of two characteristic time-scale. Such quantitative characterization is important from a cellular-level ethology point of view as it has direct connotation to their exploratory and foraging strategies. Interestingly, outside the amoebozoa and metazoa, there are largely uncharacterized species in the excavate taxon Heterolobosea including amoeboflagellate Naegleria. While classical works have shown that these cells indeed show typical amoeboid locomotion on an attached surface, their quantitative features are so far unexplored. Here, we analyzed the cell movement of Naegleria gruberi by employing long-time phase contrast imaging that automatically tracks individual cells. We show that the cells move as a persistent random walker with two time-scales that are close to those known in Dictyostelium and neutrophils. Similarities were also found in the shape dynamics which are characterized by the appearance, splitting and annihilation of the curvature waves along the cell edge. Our analysis based on the Fourier descriptor and a neural network classifier point to importance of morphology features unique to Naegleria including complex protrusions and the transient bipolar dumbbell morphologies

High-throughput analysis of spatio-temporal dynamics in Dictyostelium

A time-lapse based approach is presented that allows a rapid examination of the spatio-temporal dynamics of Dictyostelium cell populations, enabling users to search and retrieve movies on a gene-by-gene and phenotype-by-phenotype basis

Phosphatidic acid-dependent localization and basal de-phosphorylation of RA-GEFs regulate lymphocyte trafficking

Background: Lymphocytes circulate between peripheral lymphoid tissues via blood and lymphatic systems, and chemokine-induced migration is important in trafficking lymphocytes to distant sites. The small GTPase Rap1 is important in mediating lymphocyte motility, and Rap1-GEFs are involved in chemokine-mediated Rap1 activation. Here, we describe the roles and mechanisms of Rap1-GEFs in lymphocyte trafficking. Results: In this study, we show that RA-GEF-1 and 2 (also known as Rapgef2 and 6) are key guanine nucleotide exchange factors (GEF) for Rap1 in lymphocyte trafficking. Mice harboring T cell-specific knockouts of Rapgef2/6 demonstrate defective homing and egress of T cells. Sphingosine-1-phosphate (S1P) as well as chemokines activates Rap1 in a RA-GEF-1/2-dependent manner, and their deficiency in T cells impairs Mst1 phosphorylation, cell polarization, and chemotaxis toward S1P gradient. On the other hand, B cell-specific knockouts of Rapgef2/6 impair chemokine-dependent retention of B cells in the bone marrow and passively facilitate egress. Phospholipase D2-dependent production of phosphatidic acid by these chemotactic factors determines spatial distribution of Rap1-GTP subsequent to membrane localization of RA-GEFs and induces the development of front membrane. On the other hand, basal de-phosphorylation of RA-GEFs is necessary for chemotactic factor-dependent increase in GEF activity for Rap1. Conclusions: We demonstrate here that subcellular distribution and activation of RA-GEFs are key factors for a directional movement of lymphocytes and that phosphatidic acid is critical for membrane translocation of RA-GEFs with chemokine stimulation

Effect of Extraction Variables on the Biodegradable Chelant-Assisted Removal of Toxic Metals from Artificially Contaminated European Reference Soils

Development of aminopolycarboxylate chelants (APCs) having enhanced biodegradability is gaining increasing focus to replace the EDTA and its homologs with those used widely for the ex situ treatment of contaminated soils and are potential eco-threats. The paper reports the chelant-assisted extraction of the toxic metals (Cd, Cu, Pb, and Zn) from the metal-spiked European reference soils (Eurosoil 1 and Eurosoil 4) using biodegradable APCs, namely EDDS, GLDA, and HIDS. The effects of chelant-to-metal molar ratio, solution pH, and metal/chelant stability constants were evaluated, and compared with that of EDTA. The selectivity aptitude of the biodegradable chelants towards the toxic metals was assumed from the speciation calculations, and a proportionate correlation was observed at neutral pH. Pre- and post-extractive solid phase distributions of the target metals were defined using the sequential extraction procedure and dissolution of metals from the theoretically immobilized fraction was witnessed. The effect of competing species (Al, Ca, Fe, Mg, and Mn) concentrations was proven to be minimized with an excess of chelant in solution. The highlight of the outcomes is the superior decontamination ability of GLDA, a biodegradable APC, at minimum chelant concentration in solution and applicability at a wide range of pH environments. © 2012 Springer Science+Business Media Dordrecht

Selective recovery of indium from lead-smelting dust

Non-ferrous smelting dust, especially lead-smelting dust (LSD), contains percent levels of indium and thus constitutes a novel indium resource. The main difficulty in recovering indium from LSD is the coexisting presence of lead and zinc. In this study, a unique indium separation process was designed, combining techniques that involve washing with a chelant, leaching with acid and precipitation as hydroxide. The majority of the Pb in the LSD was selectively separated during chelant-assisted washing with ethylenediaminedisuccinate (EDDS), while the residual Pb was diminished through an acid leaching treatment with a mixed solution of sulfuric acid and hydrochloric acid. The chelant washing step also ensures a decrease in the raw LSD weight at a ratio of approximately 82% due to the removal of lead and counterions such as sulfate, and the washing step also minimizes the consumption of corrosive acids in the subsequent step. Selective indium separation from LSD is further complicated by the similarity of the behavior of zinc during the acid leaching step. Therefore, hydroxide precipitation at pH 5 has been introduced as the final step, ensuring the maintenance of zinc as a soluble species in the supernatant and the selective separation of indium (~88%) as a hydroxide precipitate. © 2015 Elsevier B.V.Embargo Period 48mouth

Selective recovery of indium from the etching waste solution of the flat-panel display fabrication process

The waste byproducts from the indium-consuming fabrication processes are considered as the viable resource for indium due to the unique preference to the element in designing optoelectronic devices. The present work introduces a new technique for the selective recovery of indium from the etching waste, which produced during the patterning of indium tin oxide (ITO) layer on the flat-panel displays. The process includes the application of a solid phase extraction (SPE) assembly, known as molecular recognition technology (MRT) gel, consisting of a metal-selective ligand immobilized to silica gel or polymer substrates. The samples were the real solution of etching waste from the liquid-crystal display fabrication process, and the simulated waste solution prepared using the commercially available etching solution composition containing ethanedioic acid. The retention and the corresponding indium recovery rate (%) were the key characteristics for the appraisal of MRT-SPE types: AnaLig TE 02 (TE 02), AnaLig TE 03, AnaLig TE 07, AnaLig TE 13 and AnaLig PM 02. The other conclusive factors were the influence of ion intensity in solution and the interfering effects from tin, the co-content element in the ITO-formulation. The TE 02 MRT-SPE came across all the requisite aspects for the selective recovery of indium from the etching effluent. The indium retention or recovery with TE 02 MRT-SPE from the real etching waste solution was in the range of 97 to 99% with the relative standard deviation of <4.4%. The separation of the ITO co-element tin from the waste mixture was successively selective, and thereby minimizes the possibility of interference. A three-step elution with 0.3molL-1 HNO3, 6molL-1 HCl and 1molL-1 HCl/10mmolL-1 EDTA was required to accomplish the sequential selectivity in the process. The indium retention capacity of the TE 02 MRT-SPE was 0.147mmolg-1. © 2013 Elsevier B.V