A high-throughput screening method for determining the substrate scope of nitrilases

Nitrile compounds are intermediates in the synthesis of pharmaceuticals such as atorvastatin. We have developed a chromogenic reagent to screen for nitrilase activity as an alternative to Nessler's reagent. It produces a semi-quantifiable blue colour and hydrolysis of 38 nitrile substrates by 23 nitrilases as cell-free extracts has been shown

Exploration of the role of CIB1 in cell survival and tumor growth

CIB1 is an intracellular protein with diverse functions in cancer cell biology. Here I explore two important functions of CIB1: 1) The role of CIB1 in cell survival and tumor growth in triple negative breast cancer; and 2) The interaction between CIB1 and α-integrin cytoplasmic tails and the role of this interaction in cell biology. Triple negative breast cancer (TNBC) is an aggressive breast cancer subtype with an unmet need for novel targeted therapeutics. We previously demonstrated that CIB1 is necessary for cancer cell survival and proliferation via regulation of two oncogenic signaling pathways, RAS-RAF-MEK-ERK and PI3K-AKT. Because these pathways are often upregulated in TNBC, we hypothesized that CIB1 may play a broader role in TNBC cell survival and tumor growth. Here we find that CIB1 is necessary for cell survival in multiple TNBC cell lines in vitro, and TNBC xenograft tumor growth in vivo. Furthermore, elevated AKT activation status and low PTEN expression were key predictors of sensitivity to CIB1 depletion in TNBC cells. Importantly, CIB1 knockdown caused dramatic shrinkage of MDA-MB-468 xenograft tumors in vivo. RNA sequence analysis showed that CIB1 depletion activates gene programs associated with decreased proliferation and increased cell death. Taken together, our data are consistent with the emerging theory of non-oncogene addiction, where a large subset of TNBCs depend on CIB1 for cell survival and tumor growth, independent of CIB1 expression levels. Integrins are heterodimeric transmembrane receptors important for cell adhesion to the extracellular matrix and transmission of signals through the cell membrane. Integrins are regulated via binding of cytoplasmic proteins to the integrin cytoplasmic tail. CIB1 was discovered as a binding partner of one integrin, αIIb, but the role of CIB1 in binding and regulating additional integrins was previously unknown. Here I also present evidence that CIB1 interacts with integrin αV via residues in the transmembrane portion of the integrin, can access these residues in the presence of a cell membrane, and that CIB1-integrin binding may contribute to cell signaling and proliferation. In summary, CIB1 is a diverse protein with important functions in TNBC cell survival and proliferation, as well as integrin biology.Doctor of Philosoph

Evaluation of Relative Sensitivity of SAW and Flexural Plate Wave Devices for Atmospheric Sensing

The objective of this project is to evaluate the suitability of the ultrasonic flexural plate wave (FPW) device as the detector in a gas chromatograph (GC). Of particular interest is the detection of nitrous oxide (N2O). From experimental results we conclude analyte detection is achieved through two mechanisms: changes in gas density, and mass loading of the device membrane due to the sorption of gas molecules. Reducing the dead volume of the FPW chamber increased the FPW response. A comparison of the FPW response to that of the surface acoustic wave (SAW) detector provided with the GC (made by MSI, Microsensor Technologies, Inc.), shows that for unseparated N2O in N2, the FPW exhibits a sensitivity that is at least 550 times greater than that of the SAW device. A Porapak Q column was found to separate N2O from its carrier gas, N2 or He. With the Porapak Q column, a coated FPW detected 1 ppm N2O in N2 or He, with a response magnitude of 7 Hz. A coated SAW exhibited a response of 25 Hz to pure N2O. The minimal detectable N2O concentrations of the sensors were not evaluated

The Value of Removing Daily Obstacles via Everyday Problem-Solving Theory: Developing an Applied Novel Procedure to Increase Self-Efficacy for Exercise

The objective of the study was to develop a novel procedure to increase self-efficacy for exercise. Gains in one’s ability to resolve day-to-day obstacles for entering an exercise routine were expected to cause an increase in self-efficacy for exercise. Fifty-five sedentary participants (did not exercise regularly for at least 4 months prior to the study) who expressed an intention to exercise in the near future were selected for the study. Participants were randomly assigned to one of three conditions: (1) an Experimental Group in which they received a problem-solving training session to learn new strategies for solving day-to-day obstacles that interfere with exercise, (2) a Control Group with Problem-Solving Training which received a problem-solving training session focused on a typical day-to-day problem unrelated to exercise, or (3) a Control Group which did not receive any problem-solving training. Assessment of obstacles to exercise and perceived self-efficacy for exercise were conducted at baseline; perceived self-efficacy for exercise was reassessed post-intervention (1 week later). No differences in perceived challenges posed by obstacles to exercise or self-efficacy for exercise were observed across groups at baseline. The Experimental Group reported greater improvement in self-efficacy for exercise compared to the Control Group with Training and the Control Group. Results of this study suggest that a novel procedure that focuses on removing obstacles to intended planned fitness activities is effective in increasing self-efficacy to engage in exercise among sedentary adults. Implications of these findings for use in applied settings and treatment studies are discussed

Supreme Court Opinions and Audiences

This Article evaluates different rhetorical strategies Supreme Court justices employ in writing their opinions for specific audiences. Black, Owens, Wedeking, and Wohlfarth suggest justices keep lower federal courts, state governments, federal bureaucratic agencies, and the public in mind when crafting decisions, particularly to ensure compliance with the decision and avoid non-compliance. The Article identifies opinion clarity as a means of ensuring lower federal courts will follow precedent, as well as a way for smaller and less sophisticated bureaucratic agencies to avoid shirking the Court’s rulings. The Article concludes judicial clarity is only one of an arsenal of rhetorical devices used by the Supreme Court justices, and further evaluation and research may be helpful

Psychological Functioning in Adulthood: A Self-Efficacy Analysis

In the first edition of this handbook, we laid the foundation for a self-efficacy approach to understanding learning in adulthood. We examined self-efficacy applications to learning in adulthood from two broad-based theoretical perspectives: KAPA (knowledge and appraisal personality architecture; Cervone, 2004a) and SOC (selective optimization with compensation, Baltes, Lindenberger, & Staudinger, 2006). Both perspectives emphasize the dynamic interplay between dispositional, motivational, situational, and developmental contexts for successful functioning and adaptation in life. In this edition, we build upon earlier claims with new evidence regarding the central role of self-efficacy to adult development, aging, and well-being in memory, health, work, and everyday problem-solving contexts. Of these, the work context is new in this edition, and the sections on memory, problem solving, and health are expanded and updated.The unifying theme of our chapter is the individual\u27s ability to adapt flexibly to new learning opportunities that arise in adulthood and old age by relying on perceived self-efficacy as a coping resource for navigating the changing social, cognitive, and physical landscape of late adulthood

Gene expression patterns that predict sensitivity to epidermal growth factor receptor tyrosine kinase inhibitors in lung cancer cell lines and human lung tumors

BACKGROUND: Increased focus surrounds identifying patients with advanced non-small cell lung cancer (NSCLC) who will benefit from treatment with epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI). EGFR mutation, gene copy number, coexpression of ErbB proteins and ligands, and epithelial to mesenchymal transition markers all correlate with EGFR TKI sensitivity, and while prediction of sensitivity using any one of the markers does identify responders, individual markers do not encompass all potential responders due to high levels of inter-patient and inter-tumor variability. We hypothesized that a multivariate predictor of EGFR TKI sensitivity based on gene expression data would offer a clinically useful method of accounting for the increased variability inherent in predicting response to EGFR TKI and for elucidation of mechanisms of aberrant EGFR signalling. Furthermore, we anticipated that this methodology would result in improved predictions compared to single parameters alone both in vitro and in vivo. RESULTS: Gene expression data derived from cell lines that demonstrate differential sensitivity to EGFR TKI, such as erlotinib, were used to generate models for a priori prediction of response. The gene expression signature of EGFR TKI sensitivity displays significant biological relevance in lung cancer biology in that pertinent signalling molecules and downstream effector molecules are present in the signature. Diagonal linear discriminant analysis using this gene signature was highly effective in classifying out-of-sample cancer cell lines by sensitivity to EGFR inhibition, and was more accurate than classifying by mutational status alone. Using the same predictor, we classified human lung adenocarcinomas and captured the majority of tumors with high levels of EGFR activation as well as those harbouring activating mutations in the kinase domain. We have demonstrated that predictive models of EGFR TKI sensitivity can classify both out-of-sample cell lines and lung adenocarcinomas. CONCLUSION: These data suggest that multivariate predictors of response to EGFR TKI have potential for clinical use and likely provide a robust and accurate predictor of EGFR TKI sensitivity that is not achieved with single biomarkers or clinical characteristics in non-small cell lung cancers