Inhibition of 2-oxoglutarate dependent oxygenases.

2-Oxoglutarate (2OG) dependent oxygenases are ubiquitous iron enzymes that couple substrate oxidation to the conversion of 2OG to succinate and carbon dioxide. In humans their roles include collagen biosynthesis, fatty acid metabolism, DNA repair, RNA and chromatin modifications, and hypoxic sensing. Commercial applications of 2OG oxygenase inhibitors began with plant growth retardants, and now extend to a clinically used pharmaceutical compound for cardioprotection.

Factor-inhibiting hypoxia-inducible factor (FIH) catalyses the post-translational hydroxylation of histidinyl residues within ankyrin repeat domains.

Factor-inhibiting hypoxia-inducible factor (FIH) is an Fe(II)/2-oxoglutarate-dependent dioxygenase that acts as a negative regulator of the hypoxia-inducible factor (HIF) by catalysing β-hydroxylation of an asparaginyl residue in its C-terminal transcriptional activation domain (CAD). In addition to the hypoxia-inducible factor C-terminal transcriptional activation domain (HIF-CAD), FIH also catalyses asparaginyl hydroxylation of many ankyrin repeat domain-containing proteins, revealing a broad sequence selectivity. However, there are few reports on the selectivity of FIH for the hydroxylation of specific residues.

Asparagine and aspartate hydroxylation of the cytoskeletal ankyrin family is catalyzed by factor-inhibiting hypoxia-inducible factor.

Factor-inhibiting hypoxia-inducible factor (FIH) catalyzes the β-hydroxylation of an asparagine residue in the C-terminal transcriptional activation domain of the hypoxia inducible factor (HIF), a modification that negatively regulates HIF transcriptional activity. FIH also catalyzes the hydroxylation of highly conserved Asn residues within the ubiquitous ankyrin repeat domain (ARD)-containing proteins. Hydroxylation has been shown to stabilize localized regions of the ARD fold in the case of a three-repeat consensus ankyrin protein, but this phenomenon has not been demonstrated for the extensive naturally occurring ARDs.

Structural and mechanistic studies on γ-butyrobetaine hydroxylase.

The final step in carnitine biosynthesis is catalyzed by γ-butyrobetaine (γBB) hydroxylase (BBOX), an iron/2-oxoglutarate (2OG) dependent oxygenase. BBOX is inhibited by trimethylhydrazine-propionate (THP), a clinically used compound. We report structural and mechanistic studies on BBOX and its reaction with THP.

Improving the safety and effectiveness of parenteral nutrition: results of a quality improvement collaboration.

Parenteral nutrition (PN) is a high-risk nutrition support modality. This article describes the approach taken by 1 hospital to improve safety and quality of this therapy as well as the challenges and obstacles to success. Process improvement strategies included revisions to the PN order form, education of clinicians (including physicians), increased collaboration between pharmacists and registered dietitians, and initiation of PN rounds during which PN patients were reviewed by the rounding team twice weekly.

Mutation analysis of HIF prolyl hydroxylases (PHD/EGLN) in individuals with features of phaeochromocytoma and renal cell carcinoma susceptibility.

Germline mutations in the von Hippel-Lindau disease (VHL) and succinate dehydrogenase subunit B (SDHB) genes can cause inherited phaeochromocytoma and/or renal cell carcinoma (RCC). Dysregulation of the hypoxia-inducible factor (HIF) transcription factors has been linked to VHL and SDHB-related RCC; both HIF dysregulation and disordered function of a prolyl hydroxylase domain isoform 3 (PHD3/EGLN3)-related pathway of neuronal apoptosis have been linked to the development of phaeochromocytoma. The 2-oxoglutarate-dependent prolyl hydroxylase enzymes PHD1 (EGLN2), PHD2 (EGLN1) and PHD3 (EGLN3) have a key role in regulating the stability of HIF-α subunits (and hence expression of the HIF-α transcription factors).

Building bridges: an innovative academic-service partnership.

Academic and service entities can create sustainable models of collaboration that realign relationships and resources to enhance the critical competencies essential for entry into practice. Our hospital and school collaborated to implement an innovative academic-service partnership model in which the academic and the care delivery enterprises intersected to accomplish goals of mutual interest and enhance and enrich the practice environment. Two existing resources-the staff nurse and faculty-were used to realign the educational process with the realities of nursing practice in an interpretive, contextual, live unfolding clinical environment that was relevant and memorable.

Structural studies on human 2-oxoglutarate dependent oxygenases.

2-Oxoglutarate and ferrous iron-dependent oxygenases have emerged as an important family of human enzymes that catalyse hydroxylations and related demethylation reactions. Their substrates in humans include proteins, nucleic acids, lipids and small molecules. They play roles in collagen biosynthesis, hypoxic sensing, regulation of gene expression and lipid biosynthesis/metabolism.

Structural basis for binding of cyclic 2-oxoglutarate analogues to factor-inhibiting hypoxia-inducible factor.

Aromatic analogues of the 2-oxoglutarate co-substrate of the hypoxia-inducible factor hydroxylases are shown to bind at the active site iron: Pyridine-2,4-dicarboxylate binds as anticipated with a single molecule chelating the iron in a bidentate manner. The binding mode of a hydroxamic acid analogue, at least in the crystalline state, is unusual because two molecules of the inhibitor are observed at the active site and partial displacement of the iron binding aspartyl residue was observed.

Crystal Structure of the 2-Oxoglutarate- and Fe(II)-Dependent Lysyl Hydroxylase JMJD6.

Lysyl and prolyl hydroxylations are well-known post-translational modifications to animal and plant proteins with extracellular roles. More recent work has indicated that the hydroxylation of intracellular animal proteins may be common. JMJD6 catalyses the iron- and 2-oxoglutarate-dependent hydroxylation of lysyl residues in arginine-serine-rich domains of RNA-splicing-related proteins.

Crystal structure of the 2-oxoglutarate- and Fe(II)-dependent lysyl hydroxylase JMJD6.

Lysyl and prolyl hydroxylations are well-known post-translational modifications to animal and plant proteins with extracellular roles. More recent work has indicated that the hydroxylation of intracellular animal proteins may be common. JMJD6 catalyses the iron- and 2-oxoglutarate-dependent hydroxylation of lysyl residues in arginine-serine-rich domains of RNA splicing-related proteins.

Invariance test of the Multidimensional Body Self-Relations Questionnaire: do women with breast cancer interpret this measure differently?

Purpose: To examine whether the meaning and interpretation of body image are similar for breast cancer survivors and women without breast cancer.

Method: Women completed the Multidimensional Body Self-Relations Questionnaire--Appearance Scales as part of two studies. There were 469 women with breast cancer and 385 women without breast cancer.

Sources of stress for breast cancer survivors involved in dragon boating: examining associations with treatment characteristics and self-esteem.

Objectives: This study sought to (1) identify common stressors faced by breast cancer survivors involved in dragon boating, (2) examine the conceptual and statistical factor groupings of the stressors, (3) identify differences in stressor factors based on treatment characteristics, and (4) examine the associations between stressor factors and two indicators of self-esteem.

Methods: Survivors (n = 470) involved in dragon boating completed a survey assessing stressor frequency, stressor intensity, stressor valence, physical self-worth, and global self-esteem, along with demographic and cancer treatment information.

Results: An exploratory factor analyses (EFA) using maximum likelihood extraction with oblique rotation revealed a four-factor solution that included physical, emotional, social, and exercise-related stressors.

Susceptibility of HIV-1 to tipranavir and other antiretroviral agents in treatment-experienced patients: The UTILIZE Study.

Objectives: The primary objective was to assess HIV-1 susceptibility to the protease inhibitor (PI) tipranavir and other antiretroviral (ARV) agents among treatment-experienced patients (TEP). Secondarily, clinicians' use of resistance testing was examined.

Methods: UTILIZE was an observational study conducted at 40 sites in the United States.

DNA zip codes control an ancient mechanism for gene targeting to the nuclear periphery.

Many genes in Saccharomyces cerevisiae are recruited to the nuclear periphery after transcriptional activation. We have identified two gene recruitment sequences (GRS I and II) from the promoter of the INO1 gene that target the gene to the nuclear periphery. These GRSs function as DNA zip codes and are sufficient to target a nucleoplasmic locus to the nuclear periphery.

Selective inhibitors of the JMJD2 histone demethylases: combined nondenaturing mass spectrometric screening and crystallographic approaches.

Ferrous ion and 2-oxoglutarate (2OG) oxygenases catalyze the demethylation of N(epsilon)-methylated lysine residues in histones. Here we report studies on the inhibition of the JMJD2 subfamily of histone demethylases, employing binding analyses by nondenaturing mass spectrometry (MS), dynamic combinatorial chemistry coupled to MS, turnover assays, and crystallography. The results of initial binding and inhibition assays directed the production and analysis of a set of N-oxalyl-d-tyrosine derivatives to explore the extent of a subpocket at the JMJD2 active site.

Crystal structure of the PHF8 Jumonji domain, an Nepsilon-methyl lysine demethylase.

Crystallographic analysis of the catalytic domain of PHD finger protein 8 (PHF8), an N(epsilon)-methyl lysine histone demethylase associated with mental retardation and cleft lip/palate, reveals a double-stranded beta-helix fold with conserved Fe(II) and cosubstrate binding sites typical of the 2-oxoglutarate dependent oxygenases. The PHF8 active site is highly conserved with those of the FBXL10/11demethylases, which are also selective for the di-/mono-methylated lysine states, but differs from that of the JMJD2 demethylases which are selective for tri-/di-methylated states. The results rationalize the lack of activity for the clinically observed F279S PHF8 variant and they will help to identify inhibitors selective for specific N(epsilon)-methyl lysine demethylase subfamilies.

Crystallographic and mass spectrometric analyses of a tandem GNAT protein from the clavulanic acid biosynthesis pathway.

(3R,5R)-Clavulanic acid (CA) is a clinically important inhibitor of Class A beta-lactamases. Sequence comparisons suggest that orf14 of the clavulanic acid biosynthesis gene cluster encodes for an acetyl transferase (CBG). Crystallographic studies reveal CBG to be a member of the emerging structural subfamily of tandem Gcn5-related acetyl transferase (GNAT) proteins.

The Posttraumatic Growth Inventory: an examination of the factor structure and invariance among breast cancer survivors.

Objective: The present study tested the proposed five-factor structure and invariance of the Posttraumatic Growth Inventory (PTGI; Tedeschi & Calhoun, 1996) in a sample of physically active breast cancer survivors.

Methods: A sample of breast cancer survivors (N=470, Mage=57.3, SD=7.

Intermethod reliability of real-time versus delayed videotaped evaluation of a high-fidelity medical simulation septic shock scenario.

Objectives: High-fidelity medical simulation (HFMS) is increasingly utilized in resident education and evaluation. No criterion standard of assessing performance currently exists. This study compared the intermethod reliability of real-time versus videotaped evaluation of HFMS participant performance.

Parents' perceptions of child-to-parent socialization in organized youth sport.

The purpose of this study was to enhance understanding of how parents are socialized by their children's organized youth sport participation. Five semistructured focus groups were conducted with youth sport parents (N = 26) and analyzed using qualitative methods based on Strauss and Corbin (1998). Sixty-three underlying themes reflected parents' perceived socialization experiences resulting from their children's organized youth sport participation.