418 research outputs found

    Using Prompt Delay and Instructive Feedback to Teach Pretend Play Skills to Children with Autism Spectrum Disorder

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    Pretend play involves carrying out routines, acting out roles, referencing absent or imaginary properties of objects, or substituting one object for another. Pretend play skills emerge in typically developing children by preschool age but are often absent or delayed in children with autism spectrum disorder (ASD). In the present study, we evaluated use of prompt delay, instructive feedback, and prompt delay with instructive feedback for the acquisition and maintenance of pretend play skills with children with ASD. Throughout training, we conducted free-play probes to evaluate generalization to a naturalistic setting. The results of the current study suggest that combining the prompt-delay and instructive-feedback procedures was most efficient for most participants. However, generalization to the free-play setting was limited. When clinically acceptable generalization was not observed during free-play probes, we used video modeling, contingent reinforcement, and prompts to increase responding during free-play probes

    Kanamycin resistance during in vitro development of pollen from transgenic tomato plants

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    Effects of kanamycin on pollen germination and tube growth of pollen from non-transformed plants and from transgenic tomato plants containing a chimaeric kanamycin resistance gene were determined. Germination of pollen was not affected by the addition of kanamycin to the medium in both genotypes. Kanamycin, however, severely affected tube growth of pollen from non-transformed plants, while pollen from plants containing the chimaeric gene were less sensitive and produced significantly longer tubes at kanamycin concentrations between 200-400 mg l-1. Apparently, this resistance for kanamycin correlates with the expression of the chimaeric gene during male gametophytic development.

    Effects of nitrogen fertilization on diazotrophic activity of microorganisms associated with Sphagnum magellanicum

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    In pristine ombrotrophic Sphagnum-dominated peatland ecosystems nitrogen (N) is often a limiting nutrient, which is replenished by biological N-2 fixation and atmospheric N deposition. It is, however, unclear which impact long-term N deposition has on microbial N-2 fixing activity and diazotrophic diversity, and whether phosphorus (P) modulates the response. Therefore, we studied the impact of increased N deposition and N depletion on microbial N-2 fixation and diazotrophic diversity associated with the peat moss Sphagnum magellanicum, and their interaction with P availability.Nitrogenase activities of S. magellanicum-associated microorganisms were determined by acetylene reduction assays (ARA) and N-15(2) tracer methods on mosses from two geographically distinct locations with different N deposition histories, high or low N deposition, and in samples depleted in N (grown 3 years in the greenhouse) versus recent field samples. The short-term response to increased N deposition was tested for mosses differing in N and P fertilization histories. In addition, diversity of diazotrophic microorganisms was assessed by nifH gene amplicon sequencing of N-depleted mosses.We showed distinct and persistent differences in diazotrophic communities and their activities associated with S. magellanicum from sites with high versus low N deposition. Initially, diazotrophic activity was six times higher for the low N site. During incubation and repeated ARA, however, this activity strongly decreased, while it remained stable for the high N site. Activity for the high N site could not be increased by long-term experimental N deprivation. Short-term, experimental N application had an inhibitory effect on N-2 fixation for both sites, which was not observed in mosses with high indirect P availability.We conclude that although N deposition negatively affects N-2 fixation as also shown in previous studies, long-term effects of N deprivation on the diazotrophic activity and community are more complex. Furthermore, our results indicated that P availability might be an important factor in modulating the response of Sphagnum-associated diazotrophs to N deposition.</p

    Metal ions shape α-synuclein

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    α-Synuclein is an intrinsically disordered protein that can self-aggregate and plays a major role in Parkinson’s disease (PD). Elevated levels of certain metal ions are found in protein aggregates in neurons of people suffering from PD, and environmental exposure has also been linked with neurodegeneration. Importantly, cellular interactions with metal ions, particularly Ca2+, have recently been reported as key for α-synuclein’s physiological function at the pre-synapse. Here we study effects of metal ion interaction with α-synuclein at the molecular level, observing changes in the conformational behaviour of monomers, with a possible link to aggregation pathways and toxicity. Using native nano-electrospray ionisation ion mobility-mass spectrometry (nESI-IM-MS), we characterize the heterogeneous interactions of alkali, alkaline earth, transition and other metal ions and their global structural effects on α-synuclein. Different binding stoichiometries found upon titration with metal ions correlate with their specific binding affinity and capacity. Subtle conformational effects seen for singly charged metals differ profoundly from binding of multiply charged ions, often leading to overall compaction of the protein depending on the preferred binding sites. This study illustrates specific effects of metal coordination, and the associated electrostatic charge patterns, on the complex structural space of the intrinsically disordered protein α-synuclein

    Study of the Conformational Dynamics of Prolyl Oligopeptidase by Mass Spectrometry: Lessons Learned

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    Ion mobility mass spectrometry (IM-MS) can be used to analyze native proteins according to their size and shape. By sampling individual molecules, it allows us to study mixtures of conformations, as long as they have different collision cross sections and maintain their native conformation after dehydration and vaporization in the mass spectrometer. Even though conformational heterogeneity of prolyl oligopeptidase has been demonstrated in solution, it is not detectable in IM-MS. Factors that affect the conformation in solution, binding of an active site ligand, the stabilizing Ser554Ala mutation, and acidification do not qualitatively affect the collision-induced unfolding pattern. However, measuring the protection of accessible cysteines upon ligand binding provides a principle for the development of MS-based ligand screening methods
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