An efficient temporal logic for robotic task planning

Computations required for temporal reasoning can be prohibitively expensive if fully general representations are used. Overly simple representations, such as totally ordered sequence of time points, are inadequate for use in a nonlinear task planning system. A middle ground is identified which is general enough to support a capable nonlinear task planner, but specialized enough that the system can support online task planning in real time. A Temporal Logic System (TLS) was developed during the Intelligent Task Automation (ITA) project to support robotic task planning. TLS is also used within the ITA system to support plan execution, monitoring, and exception handling

Novobiocin as an Allosteric Modulator of Ste2p

G protein-coupled receptors (GPCRs) are the target of 30-50% of all prescribed drugs for human medicine and are therefore the subject of intense study by the scientific community. It has been recognized recently that compounds called allosteric modulators can regulate GPCR activity by binding a GPCR at sites not occupied by the normal receptor-activating molecule. Such allosteric compounds are desirable drug candidates as they may produce fewer toxic side-effects than standard drugs that target GPCRs. The purpose of this study was to determine the interaction of different allosteric modulators with Ste2p, a model GPCR expressed in the yeast Saccharomyces cerevisiae. An allosteric peptide, [Bio-DOPA]11-mer, was chemically cross-linked into Ste2p in the presence and absence of another allosteric modulator, the antibiotic novobiocin. The receptor was isolated, collected, and then visualized by protein immunoblot. One of the blots detected the presence of the receptor, and the second blot detected the presence of the biotinylated ligand-receptor complex containing the cross-linked [Bio-DOPA]11-mer. Analysis of the blots revealed that the receptor was present in all of the samples and that there was significantly less [Bio-DOPA]11-mer cross-linked to the receptor in the presence of novobiocin. This experiment demonstrated that both novobiocin and [Bio-DOPA]11-mer competed for a similar site of the receptor. Thus, these two compounds that are very different in their chemical structure occupy a similar allosteric site to regulate GPCR activity. Further experimental analysis may provide insights into the mechanisms utilized by these compounds to influence GPCR function. These results may prove useful in the optimization of allosteric modulators as therapeutic agents for GPCR-based pathologies

Suing an Electronic Address: In Rem Domain Name Actions Under the ACPA

The following Article briefly examines the legal tools originally available to trademark owners (under federal law and an alternate dispute resolution procedure) and the deficiencies of each tool in anti-cybersquatting litigation. This Article presents an overview of the Anticybersquatting Consumer Protection Act (ACPA) and examines how it addresses some of the failures of prior legal tools, with particular emphasis on the ACPA\u27s in rem jurisdiction provision

UV Laser-Induced, Time-Resolved Transcriptome Responses of Saccharomyces cerevisiae

We determined the effect on gene transcription of laser-mediated, long-wavelength UV-irradiation of Saccharomyces cerevisiae by RNAseq analysis at times T15, T30, and T60 min after recovery in growth medium. Laser-irradiated cells were viable, and the transcriptional response was transient, with over 400 genes differentially expressed at T15 or T30, returning to basal level transcription by T60. Identification of transcripts exhibiting enhanced differential expression that were unique to UV laser-irradiation were identified by imposing a stringent significance cut-off (P \u3c 0.05, log2 difference \u3e2) then filtering out genes known as environmental stress response (ESR) genes. Using these rigorous criteria, 56 genes were differentially expressed at T15; at T30 differential expression was observed for 57 genes, some of which persisted from T15. Among the highly up-regulated genes were those supporting amino acid metabolic processes sulfur amino acids, methionine, aspartate, cysteine, serine), sulfur regulation (hydrogen sulfite metabolic processes, sulfate assimilation, sulfate reduction), proteasome components, amino acid transporters, and the iron regulon. At T30, the expression profile shifted to expression of transcripts related to catabolic processes (oxidoreductase activity, peptidase activity). Transcripts common to both T15 and T30 suggested an up-regulation of catabolic events, including UV damage response genes, and protein catabolism via proteasome and peptidase activity. Specific genes encoding tRNAs were among the down-regulated genes adding to the suggestion that control of protein biosynthesis was a major response to long-wave UV laser irradiation. These transcriptional responses highlight the remarkable ability of the yeast cell to respond to a UV-induced environmental insult

Harnessing Natural Diversity to Probe Metabolic Pathways

Analyses of cellular processes in the yeast Saccharomyces cerevisiae rely primarily upon a small number of highly domesticated laboratory strains, leaving the extensive natural genetic diversity of the model organism largely unexplored and unexploited. We asked if this diversity could be used to enrich our understanding of basic biological processes. As a test case, we examined a simple trait: the utilization of di/tripeptides as nitrogen sources. The capacity to import small peptides is likely to be under opposing selective pressures (nutrient utilization versus toxin vulnerability) and may therefore be sculpted by diverse pathways and strategies. Hitherto, dipeptide utilization in S. cerevisiae was solely ascribed to the activity of a single protein, the Ptr2p transporter. Using high-throughput phenotyping and several genetically diverse strains, we identified previously unknown cellular activities that contribute to this trait. We find that the Dal5p allantoate/ureidosuccinate permease is also capable of facilitating di/tripeptide transport. Moreover, even in the absence of Dal5p and Ptr2p, an additional activity—almost certainly the periplasmic asparaginase II Asp3p—facilitates the utilization of dipeptides with C-terminal asparagine residues by a different strategy. Another, as-yet-unidentified activity enables the utilization of dipeptides with C-terminal arginine residues. The relative contributions of these activities to the utilization of di/tripeptides vary among the strains analyzed, as does the vulnerability of these strains to a toxic dipeptide. Only by sampling the genetic diversity of multiple strains were we able to uncover several previously unrecognized layers of complexity in this metabolic pathway. High-throughput phenotyping facilitates the rapid exploration of the molecular basis of biological complexity, allowing for future detailed investigation of the selective pressures that drive microbial evolution.</p

Mutations affecting ligand specificity of the G-protein-coupled receptor for the Saccharomyces cerevisiae tridecapeptide pheromone

AbstractRandom mutations were generated in the G-protein-coupled receptor (Ste2p) for the tridecapeptide pheromone (α-factor) of Saccharomyces cerevisiae. These mutants were screened for variants that responded to antagonists. Because multiple mutations were detected in each mutant receptor recovered from the screen, site-directed mutagenesis was used to create single-site mutant receptors. Three receptors containing mutations F55V, S219P, and S259P were analyzed for their biological responses to various α-factor analogs and for their ligand binding profiles. Cells expressing each of the mutant receptors responded to α-factor as well as or better than wild-type cells in a growth arrest assay. In contrast, the binding of α-factor to the F55V and S219P mutant receptors was at least 10-fold reduced in comparison to wild-type receptor indicating a complex non-linear correlation between binding affinity and biological activity. Cells expressing mutant receptors responded to some normally inactive analogs in biological assays, despite the fact that these analogs had a low affinity for Ste2p. The analysis of these mutant receptors confirms previous findings that the first and sixth transmembrane regions of Ste2p are important for ligand interaction, ligand specificity, and/or receptor activation to initiate the signal transduction pathway. Changes in binding affinity of pheromone analogs to wild-type and mutant receptors indicate that residue 55 of Ste2p is involved with both ligand binding and signal transduction

A Paradigm for Peptide Hormone-GPCR Analyses

Work from our laboratories over the last 35 years that has focused on Ste2p, a G protein-coupled receptor (GPCR), and its tridecapeptide ligand α-factor is reviewed. Our work utilized the yeast Saccharomyces cerevisiae as a model system for understanding peptide-GPCR interactions. It explored the structure and function of synthetic α-factor analogs and biosynthetic receptor domains, as well as designed mutations of Ste2p. The results and conclusions are described using the nuclear magnetic resonance interrogation of synthetic Ste2p transmembrane domains (TMs), the fluorescence interrogation of agonist and antagonist binding, the biochemical crosslinking of peptide analogs to Ste2p, and the phenotypes of receptor mutants. We identified the ligand-binding domain in Ste2p, the functional assemblies of TMs, unexpected and interesting ligand analogs; gained insights into the bound α-factor structure; and unraveled the function and structures of various Ste2p domains, including the N-terminus, TMs, loops connecting the TMs, and the C-terminus. Our studies showed interactions between specific residues of Ste2p in an active state, but not resting state, and the effect of ligand activation on the dimerization of Ste2p. We show that, using a battery of different biochemical and genetic approaches, deep insight can be gained into the structure and conformational dynamics of GPCR-peptide interactions in the absence of a crystal structure

Harnessing Natural Diversity to Probe Metabolic Pathways

Analyses of cellular processes in the yeast Saccharomyces cerevisiae rely primarily upon a small number of highly domesticated laboratory strains, leaving the extensive natural genetic diversity of the model organism largely unexplored and unexploited. We asked if this diversity could be used to enrich our understanding of basic biological processes. As a test case, we examined a simple trait: the utilization of di/tripeptides as nitrogen sources. The capacity to import small peptides is likely to be under opposing selective pressures (nutrient utilization versus toxin vulnerability) and may therefore be sculpted by diverse pathways and strategies. Hitherto, dipeptide utilization in S. cerevisiae was solely ascribed to the activity of a single protein, the Ptr2p transporter. Using high-throughput phenotyping and several genetically diverse strains, we identified previously unknown cellular activities that contribute to this trait. We find that the Dal5p allantoate/ureidosuccinate permease is also capable of facilitating di/tripeptide transport. Moreover, even in the absence of Dal5p and Ptr2p, an additional activity—almost certainly the periplasmic asparaginase II Asp3p—facilitates the utilization of dipeptides with C-terminal asparagine residues by a different strategy. Another, as-yet-unidentified activity enables the utilization of dipeptides with C-terminal arginine residues. The relative contributions of these activities to the utilization of di/tripeptides vary among the strains analyzed, as does the vulnerability of these strains to a toxic dipeptide. Only by sampling the genetic diversity of multiple strains were we able to uncover several previously unrecognized layers of complexity in this metabolic pathway. High-throughput phenotyping facilitates the rapid exploration of the molecular basis of biological complexity, allowing for future detailed investigation of the selective pressures that drive microbial evolution

Quasi-experimental study designs series –Paper 9: Collecting Data from Quasi-Experimental Studies

Objective: To identify variables that must be coded when synthesizing primary studies that use quasi-experimental designs.  Study Design and Setting: All quasi-experimental (QE) designs.  Results: When designing a systematic review of QE studies potential sources of heterogeneity – both theory-based and methodological – must be identified. We outline key components of inclusion criteria for syntheses of quasi-experimental studies. We provide recommendations for coding content-relevant and methodological variables, and outlined the distinction between bivariate effect sizes and partial (i.e., adjusted) effect sizes. Designs used and controls employed are viewed as of greatest importance. Potential sources of bias and confounding are also addressed.  Conclusion: Careful consideration must be given to inclusion criteria and the coding of theoretical and methodological variables during the design phase of a synthesis of quasi-experimental studies. The success of the meta-regression analysis relies on the data available to the meta-analyst. Omission of critical moderator variables (i.e., effect modifiers) will undermine the conclusions of a meta-analysis

Conservative management of a grade V injury to an ectopic pelvic kidney following blunt trauma to the lower abdomen: a case report

<p>Abstract</p> <p>Introduction</p> <p>Ectopic pelvic kidneys represent an anatomic variant that remains clinically asymptomatic in most patients. While there is some literature to suggest that ectopic kidneys may be more predisposed to blunt trauma injuries, there are few examples to guide the management of these injuries. To our knowledge, we present the first case of a grade V renal injury to an ectopic pelvic kidney managed successfully with conservative measures.</p> <p>Case Presentation</p> <p>We present a case of grade V renal injury to an ectopic pelvic kidney in a 21 year-old African-American male. The clinical and radiographic findings are presented, along with the patient's conservative hospital course.</p> <p>Conclusion</p> <p>We suggest that management of grade V renal injuries to ectopic pelvic kidneys can be treated similarly to that of kidneys in normal anatomic position. Conservative measures may be considered in properly selected patients.</p