Induced encystment improves resistance to preservation and storage of Acanthamoeba castellanii

Several conditions that allow the preservation, storage and rapid, efficient recovery of viable Acanthamoeba castellanii organisms were investigated. The viability of trophozoites (as determined by time to confluence) significantly declined over a period of 12 months when stored at −70°C using dimethyl sulfoxide (DMSO; 5 or 10%) as cryopreservant. As A. castellanii are naturally capable of encystment, studies were undertaken to determine whether induced encystment might improve the viability of organisms under a number of storage conditions. A. castellanii cysts stored in the presence of Mg2+ at 4°C remained viable over the study period, although time to confluence was increased from approximately 8 days to approximately 24 days over the 12-month period. Storage of cysts at −70°C with DMSO (5 or 10%) or 40% glycerol, but not 80% glycerol as cryopreservants increased their viability over the 12-month study period compared with those stored at room temperature. Continued presence of Mg2+ in medium during storage had no adverse effects and generally improved recovery of viable organisms. The present study demonstrates that A. castellanii can be stored as a non-multiplicative form inexpensively, without a need for cryopreservation, for at least 12 months, but viability is increased by storage at −70°C

Molecular basis for resistance of acanthamoeba tubulins to all major classes of antitubulin compounds

Tubulin is essential to eukaryotic cells and is targeted by several antineoplastics, herbicides, and antimicrobials. We demonstrate that Acanthamoeba spp. are resistant to five antimicrotubule compounds, unlike any other eukaryote studied so far. Resistance correlates with critical amino acid differences within the inhibitor binding sites of the tubulin heterodimers

Molecular phylogenies disprove a hypothesized C4 reversion in Eragrostis walteri (Poaceae)

Background and Aims The main assemblage of the grass subfamily Chloridoideae is the largest known clade of C4 plant species, with the notable exception of Eragrostis walteri Pilg., whose leaf anatomy has been described as typical of C3 plants. Eragrostis walteri is therefore classically hypothesized to represent an exceptional example of evolutionary reversion from C4 to C3 photosynthesis. Here this hypothesis is tested by verifying the photosynthetic type of E. walteri and its classification. Methods Carbon isotope analyses were used to determine the photosynthetic pathway of several E. walteri accessions, and phylogenetic analyses of plastid rbcL and ndhF and nuclear internal transcribed spacer DNA sequences were used to establish the phylogenetic position of the species. Results Carbon isotope analyses confirmed that E. walteri is a C3 plant. However, phylogenetic analyses demonstrate that this species has been misclassified, showing that E. walteri is positioned outside Chloridoideae in Arundinoideae, a subfamily comprised entirely of C3 species. Conclusions The long-standing hypothesis of C4 to C3 reversion in E. walteri is rejected, and the classification of this species needs to be re-evaluate

Potential hazard consequences to personnel exposed to the ignition of small volumes of weakly confined stoichiometric hydrogen/air mixture

Many studies have been devoted to understanding the consequence of ignition events that could occur as a result of using hydrogen as an alternative to fossil fuels or when hydrogen is present in large scale industrial or nuclear waste sites. Little attention has however, been given to the effect of explosion in small scale operations: this could involve service work with manual handling and manipulation of gas containing packages or vessels. The purpose of this study is to begin to address this knowledge gap and report the results of an experimental program carried out to simulate the effect of localised and weakly confined small volume hydrogen explosions on personal safety. Three aspects of personal injury consequences are considered; injury from shock loading to the head/brain, skin burns and acoustic/hearing damage. It is concluded from ignition and acoustic noise exposure experiments, carried with stoichiometric hydrogen /air mixtures, that injuries arising from shock loading or burns to the skin are less likely than hearing damage. It is suggested that further work should focus on the noise exposure and hearing damage effects of small scale explosions

Influence of Grazing on Soil Microbial Communities on a Mixed Grass Prairie Ecosystem

The grazing of ungulates is the predominant use for much of the world\u27s semiarid rangelands. Grazing these lands can result in significant changes not only in the vegetation community but also in the soil physical, chemical and biological properties. Changes in soil physical and chemical properties and the plant community can potentially lead to changes in soil microbial communities which may have long-term ramifications for nutrient cycling and carbon (C) sequestration. The objective of this research was to ascertain the influence of three long-term grazing treatments on soil microbial communities

A Role for the Vacuolating Cytotoxin, VacA, in Colonization and Helicobacter pylori-Induced Metaplasia in the Stomach

Carriage of Helicobacter pylori strains producing more active (s1/i1) forms of VacA is strongly associated with gas-tric adenocarcinoma. To our knowledge, we are the first to determine effects of different polymorphic forms of VacA on inflammation and metaplasia in the mouse stomach. Bacteria producing the less active s2/i2 form of VacA colonized mice more efficiently than mutants null for VacA or producing more active forms of it, providing the first evidence of a positive role for the minimally active s2/i2 toxin. Strains producing more active toxin forms induced more severe and extensive metaplasia and in flammation in the mouse stomach than strains producing weakly active (s2/i2) toxin. We also examined the association in humans, controlling for cag PAI status. In human gastric biopsy specimens, the vacA i1 allele was strongly associated with precancerous intestinal metaplasia, with almost complete absence of intestinal metaplasia in subjects infected with i2-type strains, even in a vacA s1, cagA+ background

On the eigenvalues of Cayley graphs on the symmetric group generated by a complete multipartite set of transpositions

Given a finite simple graph \cG with $n$ vertices, we can construct the Cayley graph on the symmetric group $S_n$ generated by the edges of \cG, interpreted as transpositions. We show that, if \cG is complete multipartite, the eigenvalues of the Laplacian of \Cay(\cG) have a simple expression in terms of the irreducible characters of transpositions, and of the Littlewood-Richardson coefficients. As a consequence we can prove that the Laplacians of \cG and of \Cay(\cG) have the same first nontrivial eigenvalue. This is equivalent to saying that Aldous's conjecture, asserting that the random walk and the interchange process have the same spectral gap, holds for complete multipartite graphs.Comment: 29 pages. Includes modification which appear on the published version in J. Algebraic Combi

Microprobe Analysis in Human Pathology

This tutorial paper reviews the literature on the application of microprobe analysis to practical problems in diagnostic human pathology. The goal is to allow the reader ready access to the literature on specific clinical problems. Specimen preparation and commonly encountered artifacts are also considered. It is concluded that energy dispersive x-ray microanalysis and backscattered electron imaging are at present the most generally useful microprobe techniques for clinical work, and are no longer solely research tools. The findings often have diagnostic, therapeutic, and/or legal implications

New techniques for imaging and analyzing lung tissue.

The recent technological revolution in the field of imaging techniques has provided pathologists and toxicologists with an expanding repertoire of analytical techniques for studying the interaction between the lung and the various exogenous materials to which it is exposed. Analytical problems requiring elemental sensitivity or specificity beyond the range of that offered by conventional scanning electron microscopy and energy dispersive X-ray analysis are particularly appropriate for the application of these newer techniques. Electron energy loss spectrometry, Auger electron spectroscopy, secondary ion mass spectrometry, and laser microprobe mass analysis each offer unique advantages in this regard, but also possess their own limitations and disadvantages. Diffraction techniques provide crystalline structural information available through no other means. Bulk chemical techniques provide useful cross-checks on the data obtained by microanalytical approaches. It is the purpose of this review to summarize the methodology of these techniques, acknowledge situations in which they have been used in addressing problems in pulmonary toxicology, and comment on the relative advantages and disadvantages of each approach. It is necessary for an investigator to weigh each of these factors when deciding which technique is best suited for any given analytical problem; often it is useful to employ a combination of two or more of the techniques discussed. It is anticipated that there will be increasing utilization of these technologies for problems in pulmonary toxicology in the decades to come

Matrix controlled channel diffusion of sodium in amorphous silica

To find the origin of the diffusion channels observed in sodium-silicate glasses, we have performed classical molecular dynamics simulations of Na$_2$O--4SiO$_2$ during which the mass of the Si and O atoms has been multiplied by a tuning coefficient. We observe that the channels disappear and that the diffusive motion of the sodium atoms vanishes if this coefficient is larger than a threshold value. Above this threshold the vibrational states of the matrix are not compatible with those of the sodium ions. We interpret hence the decrease of the diffusion by the absence of resonance conditions.Comment: 5 pages, 4 figure