Detection of substrate binding motifs for morphine biosynthetic pathway intermediates in novel wound inducible (R,S)-reticuline 7-O-methyltransferase of Papaver somniferum

The benzylisoquinoline alkaloids (BIA) comprise a large and diverse group of nitrogen-containing secondary metabolites with about 2500 compounds identified in plants. BIA biosynthesis begins with the condensation of the tyrosine derived precursors dopamine and p-hydroxyphenylacetaldehyde to (S)-norcoclaurine. Subsequent regiospecific O- and N-methylations and aromatic ring hydroxylation lead to (S)-reticuline, which is the central intermediate for almost all BIAs. For morphinan alkaloid biosynthesis, (S)-reticuline undergoes an inversion of stereochemistry to (R)-reticuline, followed by C-C phenol coupling catalyzed by a unique cytochrome P450-dependent monooxygenase to yield salutaridine. The cDNA sequence of enzymes leading to (S)-reticuline, as well as those involved in the conversion of (R)-reticuline to salutaridine-7-O-acetate are already characterized. The inversion of (S)-reticuline to (R)-reticuline represent the important steps in morphine biosynthesis. Wound induced transcript accumulation in Papaver reveals a novel wound inducible EST (NCBI DbEST: GO238757) showing homology with (R,S)-reticuline 7-O-methyltransferase (ID: Q6WUC2) isolated from Papaver somniferum. We compare the substrate binding homology of this novel wound inducible (R,S)-reticuline 7-O-methyltransferase (7-OMT) using template of P. somniferum (Q6WUC2; gb|AAQ01668) as experimental control. Homology modeling with 70% identity & 85% similarity with catalytic site of template protein i.e., (Q6WUC2) short chain dehydrogenase/reductase (SDR), showed docking energy -69.9 and -75.8 kcal/mol with (S)-Reticuline (CID:439653) and (R)-Reticuline (CID:440586) respectively, which are comparable with experimental control binding site interaction energies. Docking of S- & R-reticuline into the active site revealed eight (F(5), E(18), W(24), C(47), F(44), P(45), C(46) and I(47) amino acids presumably responsible for the high substrate specificity of (R,S)-reticuline 7-O-methyltransferase

RANCH-LEVEL ECONOMIC IMPACTS OF GRAZING POLICY CHANGES: A CASE STUDY FROM OWYHEE COUNTY, IDAHO

Economic impacts often are cited as justification both for and against changes in grazing policy on public lands. A recent study conducted in Owyhee County, Idaho, illustrates a process to gather ranch-level economic information, develop economic models for different ranching systems, and use the models to estimate economic impacts of grazing policy changes. Ranch-level models were developed from producer panels and interviews within the county. Costs and returns, livestock production information, dependency on public lands, and other factors relative to ranch-level economics were gathered in four meetings with livestock producers and other interested parties. Results indicate that, as dependency on federal lands rise, both costs and returns fall. Ranch-level economic impacts of alternative grazing policy scenarios also are detailed.Land Economics/Use, Agricultural and Food Policy,

Ecuador: The Continuing Challenge of Democratic Consolidation and Civil-Military Relations; Strategic Insights, v. 5, issue 2 (February 2006)

This article appeared in Strategic Insights, v.5, issue 2 (February 2006)Approved for public release; distribution is unlimited

The Center for Civil-Military Relations / Spanish Case Study

The Center for Civil-Military Relations at the Naval Post-Graduate School (CCMR, Monterey, CA) is an implementing organization of the U.S. Department of Defense's Expanded-International Military Education and Training Program and has amassed both scholarly and practical expertise educating civilian and military defense professionals from more than 40 countries. CCMR was established in 1994 and is sponsored by the Defense Security Cooperation Agency (DSCA). CCMR conducts civil-military relations programs designed primarily for military officers, civilian officials, legislators, and non-government personnel. These programs include courses designed to be taught both in residence at NPS and in a Mobile Education Team (MET) format, depending upon requirements. Three programs offered by CCMR include the MET, the Masters Degree in International Security and Civil-Military Relations, and the Executive Program in Civil-Military Relations.The U.S. Agency for International Development (USAID) sponsored the research in this report under agreement AEP-A-00-98-00014-00, which established the Partnership for Democratic Governance and Society. The Partnership for Democratic Governance and Society (PDGS) conducts programs to strengthen the capacity of civilians to provide leadership in defense management, policymaking and analysis. The PDGS conducts its programs in cooperation with local legislatures, political parties, civic organizations, academic institutions, media and the civilian elements of the defense establishment.Approved for public release; distribution is unlimited

Portraying persons who inject drugs recently infected with hepatitis C accessing antiviral treatment : a cluster analysis

Objectives. To empirically determine a categorization of people who inject drug (PWIDs) recently infected with hepatitis C virus (HCV), in order to identify profiles most likely associated with early HCV treatment uptake. Methods.The study population was composed of HIV-negative PWIDs with a documented recent HCV infection. Eligibility criteria included being 18 years old or over, and having injected drugs in the previous 6 months preceding the estimated date of HCV exposure. Participant classification was carried out using a TwoStep cluster analysis. Results. FromSeptember 2007 to December 2011, 76 participants were included in the study. 60 participants were eligible for HCV treatment. Twenty-one participants initiated HCV treatment.The cluster analysis yielded 4 classes: class 1: Lukewarm health seekers dismissing HCV treatment offer; class 2: multisubstance users willing to shake off the hell; class 3: PWIDs unlinked to health service use; class 4: health seeker PWIDs willing to reverse the fate. Conclusion. Profiles generated by our analysis suggest that prior health care utilization, a key element for treatment uptake, differs between older and younger PWIDs. Such profiles could inform the development of targeted strategies to improve health outcomes and reduce HCV infection among PWIDs