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Filarial nematodes possess glutathione transferases (GSTs), ubiquitous enzymes with the potential to detoxify xenobiotic and endogenous substrates, and modulate the host immune system, which may aid worm infection establishment, maintenance and survival in the host. Here we have identified and characterized a σ class glycosylated GST (OoGST1), from the cattle-infective filarial nematode Onchocerca ochengi, which is homologous (99% amino acid identity) with an immunodominant GST and potential vaccine candidate from the human parasite, O. volvulus, (OvGST1b). Onchocerca ochengi native GSTs were purified using a two-step affinity chromatography approach, resolved by 2D and 1D SDS-PAGE and subjected to enzymic deglycosylation revealing the existence of at least four glycoforms. A combination of lectin-blotting and mass spectrometry (MS) analyses of the released N-glycans indicated that OoGST1 contained mainly oligomannose Man5GlcNAc2 structure, but also hybrid- and larger oligommanose-type glycans in a lower proportion. Furthermore, purified OoGST1 showed prostaglandin synthase activity as confirmed by Liquid Chromatography (LC)/MS following a coupled-enzyme assay. This is only the second reported and characterized glycosylated GST and our study highlights its potential role in host-parasite interactions and use in the study of human onchocerciasis.
Assessment of pinniped predation most often relies on analysis of the hard, undigested prey remains evident in faecal (scat) samples. For many prey species this method can yield valuable information on predator–prey interactions. For some genera, however, species diagnostic characteristics are lost during the process of prey digestion, thereby preventing morphological identification of fish prey species. Here, the feasibility of using faecal DNA to detect the presence of salmonids in pinniped scat samples and to distinguish reliably between sea trout Salmo trutta and Atlantic salmon S. salar was assessed. Novel salmonid mitochondrial DNA (mtDNA) primers were designed to amplify 162 bp of the 16S rDNA and a 327 bp section of the cytochrome b gene. Species-specific banding patterns were obtained by digestion of the cytochrome b PCR product with the restriction endonuclease AluI, and confirmed by the species-specific amplification of the 16S rDNA fragment from Atlantic salmon. Scats collected from captive grey seals Halichoerus grypus fed on known monospecific diets used to validate the PCR-RFLP assay indicated a probability of at least 95.8% (23 of 24 faecal extracts) of detecting salmonids using DNA extracted from the scat matrix. Implemented alongside conventional prey remains analyses, this technique presents a promising new method for examining prey composition and assessing pinniped predation on salmonids.
We have measured the transient events of the α-β martensitic transformation in nanocrystalline Ti films via single shot electron diffraction patterns with 1.5 ns temporal resolution. This was accomplished with a newly constructed dynamic transmission electron microscope (DTEM), which combines pulsed laser systems and pump-probe techniques with a conventional TEM. The DTEM thereby enables studies of transformations that are (1) far too fast to be captured by conventional bulk techniques, and (2) difficult to study with current ultrafast electron diffraction (UED) instruments (which typically require an accumulation of multiple shots for each diffraction pattern). Martensitic transformations in nanocrystalline materials meet both criteria, with their rapid nucleation, characteristic interface velocities ∼1 km/s, and significant irreversible microstructural changes. Free-standing 40-nm-thick Ti films were laser-heated at a rate of ∼1010 K/s to a temperature above the 1155 K transition point, then probed at various time intervals with a 1.5-ns-long intense electron pulse. Diffraction patterns show an almost complete transition to the β phase within 500 ns. Post-mortem analysis (after the sample is allowed to cool) shows a reversion to the α phase coupled with substantial grain growth, lath formation, and texture modification. The cooled material also shows a complete lack of apparent dislocations, suggesting the possible importance of a "massive" short-range diffusion mechanism.
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