Expand+Functional selection and systematic analysis of intronic splicing elements identify active sequence motifs and associated splicing factors

Despite the critical role of pre-mRNA splicing in generating proteomic diversity and regulating gene expression, the sequence composition and function of intronic splicing regulatory elements (ISREs) have not been well elucidated. Here, we employed a high-throughput in vivo Screening PLatform for Intronic Control Elements (SPLICE) to identify 125 unique ISRE sequences from a random nucleotide library in human cells. Bioinformatic analyses reveal consensus motifs that resemble splicing regulatory elements and binding sites for characterized splicing factors and that are enriched in the introns of naturally occurring spliced genes, supporting their biological relevance. In vivo characterization, including an RNAi silencing study, demonstrate that ISRE sequences can exhibit combinatorial regulatory activity and that multiple trans-acting factors are involved in the regulatory effect of a single ISRE. Our work provides an initial examination into the sequence characteristics and function of ISREs, providing an important contribution to the splicing code

Reprogramming Cellular Behavior with RNA Controllers Responsive to Endogenous Proteins

Synthetic genetic devices that interface with native cellular pathways can be used to change natural networks to implement new forms of control and behavior. The engineering of gene networks has been limited by an inability to interface with native components. We describe a class of RNA control devices that overcome these limitations by coupling increased abundance of particular proteins to targeted gene expression events through the regulation of alternative RNA splicing. We engineered RNA devices that detect signaling through the nuclear factor κB and Wnt signaling pathways in human cells and rewire these pathways to produce new behaviors, thereby linking disease markers to noninvasive sensing and reprogrammed cellular fates. Our work provides a genetic platform that can build programmable sensing-actuation devices enabling autonomous control over cellular behavior

Computing CMB Anisotropy in Compact Hyperbolic Spaces

The measurements of CMB anisotropy have opened up a window for probing the global topology of the universe on length scales comparable to and beyond the Hubble radius. For compact topologies, the two main effects on the CMB are: (1) the breaking of statistical isotropy in characteristic patterns determined by the photon geodesic structure of the manifold and (2) an infrared cutoff in the power spectrum of perturbations imposed by the finite spatial extent. We present a completely general scheme using the regularized method of images for calculating CMB anisotropy in models with nontrivial topology, and apply it to the computationally challenging compact hyperbolic topologies. This new technique eliminates the need for the difficult task of spatial eigenmode decomposition on these spaces. We estimate a Bayesian probability for a selection of models by confronting the theoretical pixel-pixel temperature correlation function with the COBE-DMR data. Our results demonstrate that strong constraints on compactness arise: if the universe is small compared to the `horizon' size, correlations appear in the maps that are irreconcilable with the observations. If the universe is of comparable size, the likelihood function is very dependent upon orientation of the manifold wrt the sky. While most orientations may be strongly ruled out, it sometimes happens that for a specific orientation the predicted correlation patterns are preferred over the conventional infinite models.Comment: 15 pages, LaTeX (IOP style included), 3 color figures (GIF) in separate files. Minor revision to match the version accepted in Class. Quantum Grav.: Proc. of Topology and Cosmology, Cleveland, 1997. The paper can be also downloaded from http://www.cita.utoronto.ca/~pogosyan/cwru_proc.ps.g

Dimension of the Torelli group for Out(F_n)

Let T_n be the kernel of the natural map from Out(F_n) to GL(n,Z). We use combinatorial Morse theory to prove that T_n has an Eilenberg-MacLane space which is (2n-4)-dimensional and that H_{2n-4}(T_n,Z) is not finitely generated (n at least 3). In particular, this recovers the result of Krstic-McCool that T_3 is not finitely presented. We also give a new proof of the fact, due to Magnus, that T_n is finitely generated.Comment: 27 pages, 9 figure

Noncyclic covers of knot complements

Hempel has shown that the fundamental groups of knot complements are residually finite. This implies that every nontrivial knot must have a finite-sheeted, noncyclic cover. We give an explicit bound, $\Phi (c)$, such that if $K$ is a nontrivial knot in the three-sphere with a diagram with $c$ crossings and a particularly simple JSJ decomposition then the complement of $K$ has a finite-sheeted, noncyclic cover with at most $\Phi (c)$ sheets.Comment: 29 pages, 8 figures, from Ph.D. thesis at Columbia University; Acknowledgments added; Content correcte

Effects of Grade Control Structures on Fish Passage, Biological Assemblages and Hydraulic Environments in Western Iowa Streams: A Multidisciplinary Review

Land use changes and channelization of streams in the deep loess region of western Iowa have led to stream channel incision, altered flow regimes, increased sediment inputs, decreased habitat diversity and reduced lateral connectivity of streams and floodplains. Grade control structures (GCSs) are built in streams to prevent further erosion, protect infrastructure and reduce sediment loads. However, GCS can have a detrimental impact on fisheries and biological communities. We review three complementary biological and hydraulic studies on the effects of GCS in these streams. GCS with steep (≥1:4 rise : run) downstream slopes severely limited fish passage, but GCS with gentle slopes (≤1:15) allowed greater passage. Fish assemblages were dominated by species tolerant of degradation, and Index of Biotic Integrity (IBI) scores were indicative of fair or poor biotic integrity. More than 50% of fish species had truncated distributions. After modification of GCS to reduce slopes and permit increased passage, IBI scores increased and several species were detected further upstream than before modification. Total macroinvertebrate density, biomass and taxonomic diversity and abundance of ecologically sensitive taxa were greater at GCS than in reaches immediately upstream, downstream or ≥1 km from GCS. A hydraulic study confirmed results from fish passage studies; minimum depths and maximum current velocities at GCS with gentle slopes (≤1:15) were more likely to meet minimum criteria for catfish passage than GCS with steeper slopes. Multidisciplinary approaches such as ours will increase understanding of GCS-associated factors influencing fish passage, biological assemblage structure and other ecological relationships in streams

Augmenting forearm crutches with wireless sensors for lower limb rehabilitation

Forearm crutches are frequently used in the rehabilitation of an injury to the lower limb. The recovery rate is improved if the patient correctly applies a certain fraction of their body weight (specified by a clinician) through the axis of the crutch, referred to as partial weight bearing (PWB). Incorrect weight bearing has been shown to result in an extended recovery period or even cause further damage to the limb. There is currently no minimally invasive tool for long-term monitoring of a patient's PWB in a home environment. This paper describes the research and development of an instrumented forearm crutch that has been developed to wirelessly and autonomously monitor a patient's weight bearing over the full period of their recovery, including its potential use in a home environment. A pair of standard forearm crutches are augmented with low-cost off-the-shelf wireless sensor nodes and electronic components to provide indicative measurements of the applied weight, crutch tilt and hand position on the grip. Data are wirelessly transmitted between crutches and to a remote computer (where they are processed and visualized in LabVIEW), and the patient receives biofeedback by means of an audible signal when they put too much or too little weight through the crutch. The initial results obtained highlight the capability of the instrumented crutch to support physiotherapists and patients in monitoring usage