In Vitro Studies of the Prp9·Prp11·Prp21 Complex Indicate a Pathway for U2 Small Nuclear Ribonucleoprotein Activation

Pre-mRNA splicing takes place on a large ribonucleoprotein particle, the spliceosome which contains the five small nuclear ribonucleoproteins (snRNPs), U1, U2, U4, U5, and U6. In Saccharomyces cerevisiae the mRNA splicing factors, Prp9, Prp11, and Prp21, are necessary for addition of the U2 snRNP to the pre-mRNA in an early step of spliceosome assembly. This paper describes a study of interactions between these proteins and their role in spliceosome assembly. The proteins were expressed in Escherichia coli. Prp9 and Prp11 were purified by metal affinity chromatography. Prp21 was purified using a solubilization/renaturation protocol. We have combined these separately purified proteins and present direct evidence of a Prp9·Prp11·Prp21 protein complex that is functional in in vitro splicing assays. Characteristics of this Prp9·Prp11·Prp21 complex were further investigated using proteins synthesized in vitro. In addition, we found that Prp9, Prp11, and Prp21 influence the structure of the U2 snRNP in a manner that alters the accessibility of the branch point pairing region of the U2 snRNA to oligonucleotide-directed RNaseH cleavage. We present a model, based on the data presented here and in the accompanying paper, for a combined role of Prp9, Prp11, Prp21, and Prp5 in activating the U2 snRNP for assembly into the pre-spliceosome

It\u27s All Local: Civil Society and the CDBG Program

This research seeks to explain contrasting patterns of population stability and decline at both the city and neighborhood levels. Existing research on urban decline identifies several factors that can facilitate or impede population stability, including: the amount of socioeconomic disadvantage, the physical and economic characteristics of a city, local government responsiveness, and the strength of civil society. Using a mixed methods approach combining multilevel modeling and spatial analysis techniques, I test the hypothesis that stronger civil society, characterized by greater collective capacity and a denser organizational resource base, is the key determinant in whether a city will stabilize, grow, or decline. First, the analysis explores divergent patterns of population change at the city level using an original time-series, cross-sectional dataset on 231 cities over four decades. At the city level, strengthening the collective capacity of civil society is an effective way to mitigate decline. Targeting local allocations of Community Development Block Grant (CDBG) funding and reducing socioeconomic disadvantage can also stimulate population growth over time. To see how these forces work at the local level, I explore neighborhood variation in Detroit, MI. The analysis looks first at local CDBG allocation decisions and then examines the forces that influence population change in Detroit neighborhoods. Geographically weighted regression results reveal significant nonstationarity in relationships between the explanatory variables and population change. In general, stronger civil society leads to greater population stability. Local CDBG investment strategies, however, tend to weaken the positive effect of civil society with a few notable exceptions. In more racially and ethnically diverse and residentially stable areas, neighborhood organizations effectively broker resources. In these neighborhoods, the combination of a strong civil society and local government investment best predict population stability. Additionally, concentrated disadvantage consistently proves the greatest obstacle to population stability or growth at the city and neighborhood levels

Thermal and elastic properties of Cu–Zr–Be bulk metallic glass forming alloys

The compositional dependence of thermal and elastic properties of Cu–Zr–Be ternary bulk metallic glass forming alloys was systematically studied. There exists a linear relationship between the glass transition temperature Tg and the total Zr concentration. G decreases linearly with increasing Zr concentration as well. The results also show that Tg, shear modulus G, and Poisson's ratio nu are very sensitive to changes in compositions. Low Tg, low G, and relatively high nu can be achieved with high Zr and Ti concentration

The Role of Occupational Therapy in Determining the Effectiveness of Functional Task Assessments Used to Improve Quality of Life for Older Adults with Parkinson’s Disease.

This critically appraised topic focuses on the role of occupational therapy in ascertaining the effectiveness of specific functional assessments in improving the quality of life of older adults with Parkinson’s disease

Prospective navigator-echo-based real-time triggering of fetal head movement for the reduction of artifacts

The purpose of this study was to evaluate the neuroimaging quality and accuracy of prospective real-time navigator-echo acquisition correction versus untriggered intrauterine magnetic resonance imaging (MRI) techniques. Twenty women in whom fetal motion artifacts compromised the neuroimaging quality of fetal MRI taken during the 28.7 ± 4week of pregnancy below diagnostic levels were additionally investigated using a navigator-triggered half-Fourier acquired single-shot turbo-spin echo (HASTE) sequence. Imaging quality was evaluated by two blinded readers applying a rating scale from 1 (not diagnostic) to 5 (excellent). Diagnostic criteria included depiction of the germinal matrix, grey and white matter, CSF, brain stem and cerebellum. Signal-difference-to-noise ratios (SDNRs) in the white matter and germinal zone were quantitatively evaluated. Imaging quality improved in 18/20 patients using the navigator echo technique (2.4 ± 0.58 vs. 3.65 ± 0.73 SD, p < 0.01 for all evaluation criteria). In 2/20 patients fetal movement severely impaired image quality in conventional and navigated HASTE. Navigator-echo imaging revealed additional structural brain abnormalities and confirmed diagnosis in 8/20 patients. The accuracy improved from 50% to 90%. Average SDNR increased from 0.7 ± 7.27 to 19.83 ± 15.71 (p < 0.01). Navigator-echo-based real-time triggering of fetal head movement is a reliable technique that can deliver diagnostic fetal MR image quality despite vigorous fetal movemen

Remodeling of brain morphology in temporal lobe epilepsy.

Mesial temporal lobe epilepsy (TLE) is one of the most widespread neurological network disorders. Computational anatomy MRI studies demonstrate a robust pattern of cortical volume loss. Most statistical analyses provide information about localization of significant focal differences in a segregationist way. Multivariate Bayesian modeling provides a framework allowing inferences about inter-regional dependencies. We adopt this approach to answer following questions: Which structures within a pattern of dynamic epilepsy-associated brain anatomy reorganization best predict TLE pathology. Do these structures differ between TLE subtypes? We acquire clinical and MRI data from TLE patients with and without hippocampus sclerosis (n = 128) additional to healthy volunteers (n = 120). MRI data were analyzed in the computational anatomy framework of SPM12 using classical mass-univariate analysis followed by multivariate Bayesian modeling. After obtaining TLE-associated brain anatomy pattern, we estimate predictive power for disease and TLE subtypes using Bayesian model selection and comparison. We show that ipsilateral para-/hippocampal regions contribute most to disease-related differences between TLE and healthy controls independent of TLE laterality and subtype. Prefrontal cortical changes are more discriminative for left-sided TLE, whereas thalamus and temporal pole for right-sided TLE. The presence of hippocampus sclerosis was linked to stronger involvement of thalamus and temporal lobe regions; frontoparietal involvement was predominant in absence of sclerosis. Our topology inferences on brain anatomy demonstrate a differential contribution of structures within limbic and extralimbic circuits linked to main effects of TLE and hippocampal sclerosis. We interpret our results as evidence for TLE-related spatial modulation of anatomical networks

Detection of local-scale population declines through optimized tidal marsh bird monitoring design

Evaluating the efficacy of monitoring designs is crucial for the successful monitoring and conservation of populations. For tidal marsh bird species of conservation concern, detecting population declines at local spatial scales within actionable time frames is a top priority. We examined and compared the effectiveness of alternative monitoring strategies for detecting local-scale population declines using count data from 1176 spatially-independent salt marsh sampling points throughout the northeastern United States (Maine to Virginia). We used abundance estimates that accounted for imperfect detection as initial conditions to simulate annual population declines of 5%, 10%, 30%, and 50% over a 5-year sampling period. Under an optimal monitoring design with biennial sampling, we were able to successfully detect annual population declines of ≥30% for each species and for all species combined. However, this required a minimum of 15–20 points per site being sampled. Power to detect declines, although low for detecting smaller annual declines (i.e., \u3c10%), improved substantially when points were visited twice per season, yet a third visit provided a reduced benefit. When testing factors that could potentially influence power to detect declines, we found that the power within sites was positively related to species abundance. Power was similar between biennial sampling (3 of 5 years) and annual sampling (5 of 5 years), suggesting a more cost-effective approach would be to sample every other year. We found that within most sites, detecting annual declines of 10% or less over a relatively short 5-year duration would be difficult. Hence, we recommend that salt marsh bird monitoring programs in the northeastern United States conduct two visits to each site per sampling year, include 15 or more sampling points per site (without confounding spatial independence), and conduct monitoring efforts every other year. This approach will maximize the efficacy of site-level monitoring of tidal marsh birds, which can aid in assessments of coastal wetland conservation and related habitat management efforts

Subversion of T lineage commitment by PU.1 in a clonal cell line system

Specification of mammalian T lymphocytes involves prolonged developmental plasticity even after lineage-specific gene expression begins. Expression of transcription factor PU.1 may maintain some myeloid-like developmental alternatives until commitment. Commitment could reflect PU.1 shutoff, resistance to PU.1 effects, and/or imposition of a suicide penalty for diversion. Here, we describe subclones from the SCID.adh murine thymic lymphoma, adh.2C2 and adh.6D4, that represent a new tool for probing these mechanisms. PU.1 can induce many adh.2C2 cells to undergo diversion to a myeloid-like phenotype, in an all-or-none fashion with multiple, coordinate gene expression changes; adh.6D4 cells resist diversion, and most die. Diversion depends on the PU.1 Ets domain but not on known interactions in the PEST or Q-rich domains, although the Q-rich domain enhances diversion frequency. Protein kinase C/MAP kinase stimulation can make adh.6D4 cells permissive for diversion without protecting from suicide. These results show distinct roles for regulated cell death and another stimulation-sensitive function that establishes a threshold for diversion competence. PU.1 also diverts normal T-cell precursors from wild type or Bcl2-transgenic mice to a myeloid-like phenotype, upon transduction in short-term culture. The adh.2C2 and adh.6D4 clones thus provide an accessible system for defining mechanisms controlling developmental plasticity in early T-cell development