Determinants of the Scope and Strength of State Environmental Policy

Using bivariate crosstabulation and multivariate regression analysis this study attempted to measure the effects of political factors on the scope and strength of state environmental policy. Political variables included state political culture, interest group strength, partisanship, gubernatorial strength, legislative professionalism, and state innovativeness. Scope and strength of state environmental policy was measured by four dependent variables. The dependent variables measured state initiated policy, federally intitiated policy, per capita spending, and toxic substance control efforts. The dependent variables measuring state initiated and federally initiated policies are new attempts in measuring state environmental policy. They are indices that combine measured characteristics of states' environmental policies in different areas of environmental concern. Overall, the four dependent variables represent a comprehensive attempt to measure environmental policy in the fifty states.Political Scienc

Perspective:Dietary Biomarkers of Intake and Exposure - Exploration with Omics Approaches

While conventional nutrition research has yielded biomarkers such as doubly labeled water for energy metabolism and 24-h urinary nitrogen for protein intake, a critical need exists for additional, equally robust biomarkers that allow for objective assessment of specific food intake and dietary exposure. Recent advances in high-throughput MS combined with improved metabolomics techniques and bioinformatic tools provide new opportunities for dietary biomarker development. In September 2018, the NIH organized a 2-d workshop to engage nutrition and omics researchers and explore the potential of multiomics approaches in nutritional biomarker research. The current Perspective summarizes key gaps and challenges identified, as well as the recommendations from the workshop that could serve as a guide for scientists interested in dietary biomarkers research. Topics addressed included study designs for biomarker development, analytical and bioinformatic considerations, and integration of dietary biomarkers with other omics techniques. Several clear needs were identified, including larger controlled feeding studies, testing a variety of foods and dietary patterns across diverse populations, improved reporting standards to support study replication, more chemical standards covering a broader range of food constituents and human metabolites, standardized approaches for biomarker validation, comprehensive and accessible food composition databases, a common ontology for dietary biomarker literature, and methodologic work on statistical procedures for intake biomarker discovery. Multidisciplinary research teams with appropriate expertise are critical to moving forward the field of dietary biomarkers and producing robust, reproducible biomarkers that can be used in public health and clinical research

Hands-on Workshops as An Effective Means of Learning Advanced Technologies Including Genomics, Proteomics and Bioinformatics

Genomics and proteomics have emerged as key technologies in biomedical research, resulting in a surge of interest in training by investigators keen to incorporate these technologies into their research. At least two types of training can be envisioned in

Co-localization of NQO1, Sirt2 and acetyl tubulin in 16HBE cells.

<p>(A) Co-immunostaining for NQO1 (green) and acetyl α-tubulin (red) showing co-localization on mitotic structures. (B) Co-immunostaining for NQO1 (green) and Sirt2 (red) showing co-localization on centrosome(s). Arrows indicate co-localization between high intensity immunostaining for NQO1, acetyl α-tubulin and Sirt2 in different stages of the centriole cycle. (C, centrosome(s); MS, mitotic spindles; MB, midbody).</p

Redox modulation of NQO1

<div><p>NQO1 is a FAD containing NAD(P)H-dependent oxidoreductase that catalyzes the reduction of quinones and related substrates. In cells, NQO1 participates in a number of binding interactions with other proteins and mRNA and these interactions may be influenced by the concentrations of reduced pyridine nucleotides. NAD(P)H can protect NQO1 from proteolytic digestion suggesting that binding of reduced pyridine nucleotides results in a change in NQO1 structure. We have used purified NQO1 to demonstrate the addition of NAD(P)H induces a change in the structure of NQO1; this results in the loss of immunoreactivity to antibodies that bind to the C-terminal domain and to helix 7 of the catalytic core domain. Under normal cellular conditions NQO1 is not immunoprecipitated by these antibodies, however, following treatment with β-lapachone which caused rapid oxidation of NAD(P)H NQO1 could be readily pulled-down. Similarly, immunostaining for NQO1 was significantly increased in cells following treatment with β-lapachone demonstrating that under non-denaturing conditions the immunoreactivity of NQO1 is reflective of the NAD(P)⁺/NAD(P)H ratio. In untreated human cells, regions with high intensity immunostaining for NQO1 co-localize with acetyl α-tubulin and the NAD⁺-dependent deacetylase Sirt2 on the centrosome(s), the mitotic spindle and midbody during cell division. These data provide evidence that during the centriole duplication cycle NQO1 may provide NAD⁺ for Sirt2-mediated deacetylation of microtubules. Overall, NQO1 may act as a redox-dependent switch where the protein responds to the NAD(P)⁺/NAD(P)H redox environment by altering its structure promoting the binding or dissociation of NQO1 with target macromolecules.</p></div

Co-localization of NQO1, Sirt2 and acetyl tubulin in 16HBE cells.

<p>(A) Co-immunostaining for NQO1 (green) and acetyl α-tubulin (red) showing co-localization on mitotic structures. (B) Co-immunostaining for NQO1 (green) and Sirt2 (red) showing co-localization on centrosome(s). Arrows indicate co-localization between high intensity immunostaining for NQO1, acetyl α-tubulin and Sirt2 in different stages of the centriole cycle. (C, centrosome(s); MS, mitotic spindles; MB, midbody).</p

Co-localization of NQO1, acetyl α-tubulin and Sirt2 in TrHBMEC.

<p>(A) Co-immunostaining for NQO1 (A180, green) and acetyl α-tubulin (red) in TrHBMEC cells showing co-localization on centrosomes (arrows). (B) Co-immunostaining for NQO1 (A180, green) and acetyl α-tubulin (red) in TrHBMEC cells showing co-localization on mitotic spindles. Co-immunostaining for NQO1 (A180, green) and Sirt2 (red) in 16HBE cells. Arrows indicate co-localization between high intensity immunostaining for NQO1, acetyl α-tubulin and Sirt2 on the centrosomes.</p

Relative positioning of the C-term and A180 antibody epitopes on human NQO1.

<p>Alternating viewpoints of the human NQO1 homodimer (PDB ID: 1D4A) with each monomer colored separately (blue and green) and the locations of the C-terminal epitopes (CT, yellow) and A180 epitopes (magenta) highlighted.</p

Intracellular oxidation of pyridine nucleotides induces a conformational change in NQO1.

<p>(A) 16HBE cells were treated with DMSO or β-lapachone (10μM) for 2 h after which NQO1 was immunoprecipitated using anti-NQO1 antibodies. (B) Human cell lines (16HBE, ARPE19, TrHBMEC) were treated with β-lapachone (10μM) for the indicated times after which NQO1 was immunoprecipitated using antibodies that recognize the C-terminal domain of NQO1. (C) Intracellular levels of NADH and NAD⁺ were measured by mass spectrometry in 16HBE cells treated with β-lapachone (10μM) for 2 h in the absence and presence of the PARP inhibitor olaparib (1μM). Results are the mean ± standard deviation, n = 3. (D) Immunoprecipitation of NQO1 from 16HBE cells treated with β-lapachone (10μM) in the absence and presence of olaparib (1μM) for 2 h. Reaction conditions are described in <i>Materials and methods</i>.</p