Intron self-complementarity enforces exon inclusion in a yeast pre-mRNA

Skipping of internal exons during removal of introns from pre-mRNA must be avoided for proper expression of most eukaryotic genes. Despite significant understanding of the mechanics of intron removal, mechanisms that ensure inclusion of internal exons in multi-intron pre-mRNAs remain mysterious. Using a natural two-intron yeast gene, we have identified distinct RNA–RNA complementarities within each intron that prevent exon skipping and ensure inclusion of internal exons. We show that these complementarities are positioned to act as intron identity elements, bringing together only the appropriate 5′ splice sites and branchpoints. Destroying either intron self-complementarity allows exon skipping to occur, and restoring the complementarity using compensatory mutations rescues exon inclusion, indicating that the elements act through formation of RNA secondary structure. Introducing new pairing potential between regions near the 5′ splice site of intron 1 and the branchpoint of intron 2 dramatically enhances exon skipping. Similar elements identified in single intron yeast genes contribute to splicing efficiency. Our results illustrate how intron secondary structure serves to coordinate splice site pairing and enforce exon inclusion. We suggest that similar elements in vertebrate genes could assist in the splicing of very large introns and in the evolution of alternative splicing.

Anandamide and diet: Inclusion of dietary arachidonate and docosahexaenoate leads to increased brain levels of the corresponding N-acylethanolamines in piglets

Endogenous ligands of cannabinoid receptors have been discovered recently and include some N-acylethanolamines (NAEs; e.g., N-arachidonoylethanolamine) and some 2-acylglycerols (e.g., sn-2-arachidonoylglycerol). Previously, we found these compounds to be active biologically when administered per os in large quantities to mice. In the present work, piglets were fed diets with and without 20:4n−6 and 22:6n−3 fatty acid precursors of NAEs, in levels similar to those found in porcine milk, during the first 18 days of life, and corresponding brain NAEs were assessed. In piglets fed diets containing 20:4n−6 and 22:6n−3, there were increases in several biologically active NAEs in brain homogenates—20:4n−6 NAE (4-fold), 20:5n−3 NAE (5-fold), and 22:5n−3 and 22:6n−3 NAE (9- to 10-fold). These results support a mechanism we propose for dietary long-chain polyunsaturated fatty acids influences on brain biochemistry with presumed functional sequelae. This paradigm will enable targeted investigations to determine whether and why specific populations such as infants, elderly, or persons suffering from certain clinical conditions may benefit from dietary long-chain polyunsaturated fatty acids.

Accumulation of Mutant Huntingtin Fragments in Aggresome-like Inclusion Bodies as a Result of Insufficient Protein Degradation

The huntingtin exon 1 proteins with a polyglutamine repeat in the pathological range (51 or 83 glutamines), but not with a polyglutamine tract in the normal range (20 glutamines), form aggresome-like perinuclear inclusions in human 293 Tet-Off cells. These structures contain aggregated, ubiquitinated huntingtin exon 1 protein with a characteristic fibrillar morphology. Inclusion bodies with truncated huntingtin protein are formed at centrosomes and are surrounded by vimentin filaments. Inhibition of proteasome activity resulted in a twofold increase in the amount of ubiquitinated, SDS-resistant aggregates, indicating that inclusion bodies accumulate when the capacity of the ubiquitin–proteasome system to degrade aggregation-prone huntingtin protein is exhausted. Immunofluorescence and electron microscopy with immunogold labeling revealed that the 20S, 19S, and 11S subunits of the 26S proteasome, the molecular chaperones BiP/GRP78, Hsp70, and Hsp40, as well as the RNA-binding protein TIA-1, the potential chaperone 14–3-3, and α-synuclein colocalize with the perinuclear inclusions. In 293 Tet-Off cells, inclusion body formation also resulted in cell toxicity and dramatic ultrastructural changes such as indentations and disruption of the nuclear envelope. Concentration of mitochondria around the inclusions and cytoplasmic vacuolation were also observed. Together these findings support the hypothesis that the ATP-dependent ubiquitin–proteasome system is a potential target for therapeutic interventions in glutamine repeat disorders.

Lipid metabolism in Chlamydia trachomatis-infected cells: directed trafficking of Golgi-derived sphingolipids to the chlamydial inclusion.

Chlamydia trachomatis undergoes its entire life cycle within an uncharacterized intracellular vesicle that does not fuse with lysosomes. We used a fluorescent Golgi-specific probe, (N-[7-(4-nitrobenzo-2-oxa-1,3-diazole)]) aminocaproylsphingosine (C6-NBD-Cer), in conjunction with conventional fluorescence or confocal microscopy to identify interactions between the Golgi apparatus and the chlamydial inclusion. We observed not only a close physical association between the Golgi apparatus and the chlamydial inclusion but the eventual presence of a metabolite of this fluorescent probe associated with the chlamydiae themselves. Sphingomyelin, endogenously synthesized from C6-NBD-Cer, was specifically transported to the inclusion and incorporated into the cell wall of the intracellular chlamydiae. Incorporation of the fluorescent sphingolipid by chlamydiae was inhibited by brefeldin A. Chlamydiae therefore occupy a vesicle distal to the Golgi apparatus that receives anterograde vesicular traffic from the Golgi normally bound for the plasma membrane. Collectively, the data suggest that the chlamydial inclusion may represent a unique compartment within the trans-Golgi network.

An inverse method to obtain porosity, fibre diameter and density of fibrous sound absorbing materials

Characterization of sound absorbing materials is essential to predict its acoustic behaviour. The most commonly used models to do so consider the flow resistivity, porosity, and average fibre diameter as parameters to determine the acoustic impedance and sound absorbing coefficient. Besides direct experimental techniques, numerical approaches appear to be an alternative to estimate the material’s parameters. In this work an inverse numerical method to obtain some parameters of a fibrous material is presented. Using measurements of the normal incidence sound absorption coefficient and then using the model proposed by Voronina, subsequent application of basic minimization techniques allows one to obtain the porosity, average fibre diameter and density of a sound absorbing material. The numerical results agree fairly well with the experimental data.

Evaluation of Two Alternative Procedures for Measuring Airflow Resistance of Sound Absorbing Materials

It is well known that sound absorption and sound transmission properties of open porous materials are highly dependent on their airflow resistance values. Low values of airflow resistance indicate little resistance for air streaming through the porous material and high values are a sign that most of the pores inside the material are closed. The laboratory procedures for measuring airflow resistance have been stan- dardized by several organizations, including ISO and ASTM for both alternate flow and continuous flow. However, practical implementation of these standardized methods could be both complex and expensive. In this work, two indirect alternative measurement procedures were compared against the alternate flow standardized technique. The techniques were tested using three families of eco-friendly sound absorbent materials: recycled polyurethane foams, coconut natural fibres, and recycled polyester fibres. It is found that the values of airflow resistance measured using both alternative methods are very similar. There is also a good correlation between the values obtained through alternative and standardized methods.

Inclusion of immersive virtual learning environments and visual control systems to support the learning of students with Asperger syndrome

This paper presents the use of immersive virtual reality systems in the educational intervention with Asperger students. The starting points of this study are features of these students' cognitive style that requires an explicit teaching style supported by visual aids and highly structured environments. The proposed immersive virtual reality system, not only to assess the student's behavior and progress, but also is able to adapt itself to the student's specific needs. Additionally, the immersive reality system is equipped with sensors that can determine certain behaviors of the students. This paper determines the possible inclusion of immersive virtual reality as a support tool and learning strategy in these particular students' intervention. With this objective two task protocols have been defined with which the behavior and interaction situations performed by participant students are recorded. The conclusions from this study talks in favor of the inclusion of these virtual immersive environments as a support tool in the educational intervention of Asperger syndrome students as their social competences and executive functions have improved.

Inclusive education in Spain: how do skills, resources, and supports affect regular education teachers’ perceptions of inclusion?

This study examined regular education teachers’ perceptions of inclusion in elementary and secondary schools in Spain and how these perceptions may differ depending on teaching experience, skills, and the availability of resources and supports. Stratified random sampling procedures were used to draw a representative sample of 336 general education teachers (68 kindergarten, 133 elementary, and 135 secondary teachers) from the province of Alicante. The results indicated the acceptance of the principles of inclusion, although teacher skills, time, material resources, and personal supports for inclusion were deemed insufficient. Kindergarten and elementary teachers showed more positive perceptions of inclusion than secondary education teachers, and so did teachers with more personal supports and material resources than those with less supports and resources. The results are discussed in terms of its implications for practice in order to promote more inclusive classrooms in Spain.

Redrawing Identity Boundaries through Integration Policies: Strategies of Inclusion/Exclusion of Immigrants in Québec and South Tyrol. EDAP Paper 01/2013

Following the research agenda introduced by Will Kymlicka, this qualitative study offers an interpretation of how the sub-national elites of Québec and South Tyrol police the integration of immigrants. For these national minority groups, which are constantly undergoing a process of redefinition of their collective identities by differentiating themselves from the Others who do not belong to the in-group, immigrants have progressively become the most significant Others as they are not part of the original system of compromises. This article questions how sub-national elites are handling this relatively new kind of ethnocultural diversity brought about by large-scale permanent immigration on two levels: first, the political narrative of the ruling sub-national parties, their electoral appeals, manifestos and speeches; second, the policy arrangements for the integration of immigrants in education, language and social policy. The initial approach of the article is pessimistic, as it assumes that sub-national elites will marginalize immigrants to please core nationalist supporters. In fact, the hypotheses to be tested are whether the national minority groups of Québec and South Tyrol engage in a process of reconstruction of their ethnic identity bounded by opposition to real or imagined Others – the newcomers; and whether they adopt practical measures that force newcomers to be assimilated into the group or to be marginalized. The comparison between Québec and South Tyrol provides a basic understanding of the impact of immigration in two sub-national polities that are very different, but still adopt similar political narratives and policy strategies with regard to the integration of newcomers.