Identifying gene locus associations with promyelocytic leukemia nuclear bodies using immuno-TRAP.

Important insights into nuclear function would arise if gene loci physically interacting with particular subnuclear domains could be readily identified. Immunofluorescence microscopy combined with fluorescence in situ hybridization (immuno-FISH), the method that would typically be used in such a study, is limited by spatial resolution and requires prior assumptions for selecting genes to probe. Our new technique, immuno-TRAP, overcomes these limitations. Using promyelocytic leukemia nuclear bodies (PML NBs) as a model, we used immuno-TRAP to determine if specific genes localize within molecular dimensions with these bodies. Although we confirmed a TP53 gene-PML NB association, immuno-TRAP allowed us to uncover novel locus-PML NB associations, including the ABCA7 and TFF1 loci and, most surprisingly, the PML locus itself. These associations were cell type specific and reflected the cell's physiological state. Combined with microarrays or deep sequencing, immuno-TRAP provides powerful opportunities for identifying gene locus associations with potentially any nuclear subcompartment

Change Detection Performance Under Divided Attention with Dynamic Driving Scenarios

This study investigated the effect of cognitive load on the relationshipbetween confidence in detecting changes and actual change detectionperformance. Two experiments simulated glancing away from the roadway byperiodically blanking the driver’s view for one second. Experiments wereconducted in a driving simulator where participants were asked to detect changesin the location and appearance of other vehicles while driving on a multi-lanesuburban roadway. In addition, cognitive load was imposed using messages thatparticipants were asked to listen to and answer questions about. Participants’sensitivity (d’) to vehicle changes was calculated and compared with subjectiveratings of confidence in detecting those changes. Results indicated a positiverelationship between d’ and confidence, suggesting that participants were aware ofthe factors that influenced their change-detection performance. However, thestrength of the relationship was situation-dependent. The strength of therelationship decreased when the detection task was more difficult and in thepresence of cognitive load

The Effect of Voice Interactions on Drivers’ Guidance of Attention

The objective of the current study was to assess the effect of voice interactions with an in-vehicle system on drivers’ guidance of attention. Our approach was to examine the effect of voice interactions on endogenous control of attention using a modified Posner cue-target paradigm. Consistent with the bottleneck hypothesis, dual-task slowing was observed when drivers responded to an auditory task and to a pedestrian detection task concurrently. This interference contributed to disrupted attention allocation, especially when drivers could not rely on their endogenous control of attention

Teaching in Kindergarten Through Grade 12 Programs: Perceptions of Native and Nonnative English-Speaking Practitioners

This study compared the perceptions of two groups of kindergarten through grade 12 (K-12) practitioners, native English-speaking teachers (NESTs) and nonnative English-speaking teachers (NNESTs), in relation to their professional preparation, their level of job satisfaction, and their degree of comfort teaching various skill areas. Quantitative and qualitative analyses of a survey administered to 55 NESTs and 32 NNESTs in Southern California showed that a complex set of similarities and differences characterized the K-12 NESTs and NNESTs who participated in this investigation. The similarities identified included the positive views of the two groups about their professional preparation and the support received from formal networks (e.g., mentoring programs) and informal networks (e.g., colleagues, friends, and relatives). The two groups were also similar in their positive self-reported level of job satisfaction and their positive perceptions of their English language skills. One of the differences was the grades they taught, with a higher percentage of NNESTs teaching early elementary grades. Additionally, NNESTs were slightly more negative than NESTs in their evaluation of school administrators, exhibited more positive self-perceptions about their instructional abilities, and saw their nonnative status as contributing to their professional abilities. The results of this preliminary investigation suggest the need for further research on NNESTs and for the development of collaborative projects between master of art (MA) degree programs in Teaching English to Speakers of Other Languages (TESOL) and K-12 programs designed to benefit future teachers, regardless of their language background

Parallel identification of O-GlcNAc-modified proteins from cell lysates

We report a new strategy for the parallel identification of O-GlcNAc-glycosylated proteins from cell lysates. The approach permits specific proteins of interest to be rapidly interrogated for the modification in any tissue or cell type and can be extended to peptides to facilitate the mapping of glycosylation sites. As an illustration of the approach, we identified four new O-GlcNAc-glycosylated proteins of low cellular abundance (c-Fos, c-Jun, ATF-1, and CBP) and two short regions of glycosylation in the enzyme O-GlcNAc transferase (OGT). The ability to target specific proteins across various tissue or cell types complements emerging proteomic technologies and should advance our understanding of this important posttranslational modification

Social and Cultural Concepts Module: Curricula to Foster Cultural Responsiveness

The lack of racial diversity in the occupational therapy profession as well as its educational programs are obstacles in providing culturally responsive care to serve all people, populations, and communities. Research has shown that student engagement in case studies, activities that emphasize exposure to diverse populations, reflective journaling, and interviewing someone from a different culture are effective in developing practitioners who are more culturally responsive. Students from one culturally diverse occupational therapy program took a nine-hour Social and Cultural Concepts Module (SCCM) that incorporated best practice pedagogy for teaching cultural responsiveness. A one group pretest-posttest design was utilized to explore how well-prepared the students believed they were to meet the needs of diverse communities following the module. Data was collected from 35 entry-level doctor of occupational therapy students from four major race/ethnicities using the Interprofessional Education Health Disparities Series Survey (IPE-HDSS). Of the 30 items measured, 22 items showed a significant positive difference from the pretest to the posttest. Results demonstrated a change in the students’ self-reported knowledge of culturally responsive care following classroom instruction. This study provides insight into the need for culturally responsive curriculum to be integral to occupational therapy education

An In Silico Approach for Evaluating a Fraction-Based, Risk Assessment Method for Total Petroleum Hydrocarbon Mixtures

Both the Massachusetts Department of Environmental Protection (MADEP) and the Total Petroleum Hydrocarbon Criteria Working Group (TPHCWG) developed fraction-based approaches for assessing human health risks posed by total petroleum hydrocarbon (TPH) mixtures in the environment. Both organizations defined TPH fractions based on their expected environmental fate and by analytical chemical methods. They derived toxicity values for selected compounds within each fraction and used these as surrogates to assess hazard or risk of exposure to the whole fractions. Membership in a TPH fraction is generally defined by the number of carbon atoms in a compound and by a compound's equivalent carbon (EC) number index, which can predict its environmental fate. Here, we systematically and objectively re-evaluate the assignment of TPH to specific fractions using comparative molecular field analysis and hierarchical clustering. The approach is transparent and reproducible, reducing inherent reliance on judgment when toxicity information is limited. Our evaluation of membership in these fractions is highly consistent (˜80% on average across various fractions) with the empirical approach of MADEP and TPHCWG. Furthermore, the results support the general methodology of mixture risk assessment to assess both cancer and noncancer risk values after the application of fractionation

Involvement of the TCR Cβ FG Loop in Thymic Selection and T Cell Function

The asymmetric disposition of T cell receptor (TCR) Cβ and Cα ectodomains creates a cavity with a side-wall formed by the rigid Cβ FG loop. To investigate the significance of this conserved structure, we generated loop deletion (βΔFG) and βwt transgenic (tg) mice using the TCR β subunit of the N15 CTL. N15βwt and N15βΔFG H-2b animals have comparable numbers of thymocytes in S phase and manifest developmental progression through the CD4−CD8− double-negative (DN) compartment. N15βΔFG facilitates transition from DN to CD4+8+ double-positive (DP) thymocytes in recombinase activating gene (RAG)-2−/− mice, showing that pre-TCR function remains. N15βΔFG animals possess ∼twofold more CD8+ single-positive (SP) thymocytes and lymph node T cells, consistent with enhanced positive selection. As an altered Vα repertoire observed in N15βΔFG mice may confound the deletion's effect, we crossed N15αβ TCR tg RAG-2−/− with N15βΔFG tg RAG-2−/− H-2b mice to generate N15αβ RAG-2−/− and N15αβ.βΔFG RAG-2−/− littermates. N15αβ.βΔFG RAG-2−/− mice show an 8–10-fold increase in DP thymocytes due to reduced negative selection, as evidenced by diminished constitutive and cognate peptide-induced apoptosis. Compared with N15αβ, N15αβ.βΔFG T cells respond poorly to cognate antigens and weak agonists. Thus, the Cβ FG loop facilitates negative selection of thymocytes and activation of T cells

FGF Signaling Regulates the Number of Posterior Taste Papillae by Controlling Progenitor Field Size

The sense of taste is fundamental to our ability to ingest nutritious substances and to detect and avoid potentially toxic ones. Sensory taste buds are housed in papillae that develop from epithelial placodes. Three distinct types of gustatory papillae reside on the rodent tongue: small fungiform papillae are found in the anterior tongue, whereas the posterior tongue contains the larger foliate papillae and a single midline circumvallate papilla (CVP). Despite the great variation in the number of CVPs in mammals, its importance in taste function, and its status as the largest of the taste papillae, very little is known about the development of this structure. Here, we report that a balance between Sprouty (Spry) genes and Fgf10, which respectively antagonize and activate receptor tyrosine kinase (RTK) signaling, regulates the number of CVPs. Deletion of Spry2 alone resulted in duplication of the CVP as a result of an increase in the size of the placode progenitor field, and Spry1−/−;Spry2−/− embryos had multiple CVPs, demonstrating the redundancy of Sprouty genes in regulating the progenitor field size. By contrast, deletion of Fgf10 led to absence of the CVP, identifying FGF10 as the first inductive, mesenchyme-derived factor for taste papillae. Our results provide the first demonstration of the role of epithelial-mesenchymal FGF signaling in taste papilla development, indicate that regulation of the progenitor field size by FGF signaling is a critical determinant of papilla number, and suggest that the great variation in CVP number among mammalian species may be linked to levels of signaling by the FGF pathway