DXP Synthase‐Catalyzed C?N Bond Formation: Nitroso Substrate Specificity Studies Guide Selective Inhibitor Design

1‐Deoxy‐D ‐xylulose 5‐phosphate (DXP) synthase catalyzes the first step in the nonmammalian isoprenoid biosynthetic pathway to form DXP from pyruvate and D ‐glyceraldehyde 3‐phosphate (D ‐GAP) in a thiamin diphosphate‐dependent manner. Its unique structure and mechanism distinguish DXP synthase from its homologues and suggest that it should be pursued as an anti‐infective drug target. However, few reports describe any development of selective inhibitors of this enzyme. Here, we reveal that DXP synthase catalyzes C? N bond formation and exploit aromatic nitroso substrates as active site probes. Substrate specificity studies reveal a high affinity of DXP synthase for aromatic nitroso substrates compared to the related ThDP‐dependent enzyme pyruvate dehydrogenase (PDH). Results from inhibition and mutagenesis studies indicate that nitroso substrates bind to E. coli DXP synthase in a manner distinct from that of D ‐GAP. Our results suggest that the incorporation of aryl acceptor substrate mimics into unnatural bisubstrate analogues will impart selectivity to DXP synthase inhibitors. As a proof of concept, we show selective inhibition of DXP synthase by benzylacetylphosphonate (BnAP).     

Experimental Design in the Optimization of a Microwave Acid Digestion Procedure of Biomorphic Ceramic Based on ZrO2

Abstract

The present work deals with the manufacturing of biomorphous ZrO₂-ceramics from oak wood as biological template structure. Oak wood was vacuum infiltrated with zirconia-sol. Subsequent pyrolysis in inert atmosphere at 800°C and annealing in air up to 1550°C resulted in the formation of porous, microcellular ZrO₂-ceramics. After the material characterization, we optimized the sample dissolution by acid attack in an oven under microwave irradiation. Experimental designs were used as a multivariate strategy for the effects evaluation of varying several variables. The optimization was performed using full factorial design 2⁴. Four variables (time, power, volume of HNO₃, and volume of HF) were considered as factors and as response the concentration of different metal ions in the optimization process.     

Distribution of mono‐ and di‐methacrylic monomers in SBS block copolymer and its influence on the photopolymerization process

Persoz hardness measurements and the analysis of the temperature dependence of the storage modulus (E′) for SBS di‐ and tetrafunctional methacrylic monomer systems were carried out to determine the distribution of the monomers in the domains or phases of the SBS block copolymer, as well as the aggregation state of each system. The forces of attraction between the monomer and the phases or domains in the matrix (similar solubility parameters with appreciable dipole‐dipole and hydrogen bonding interactions) were determinant as for the monomer distribution. The influence of these structural factors on the kinetic parameters of the photoinitiated polymerization of di‐ and tetrafunctional methacrylic monomers in the SBS matrix was studied. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 98: 163–168, 2005     

BMS Derivatives C7-Linked to β-Cyclodextrin and Hyperbranched Polyglycerol Retain Activity against R5-HIV-1NLAD8 Isolates and Can Be Deemed Potential Microbicides

Amides from indole-3-glyoxylic acid and 4-benzoyl-2-methylpiperazine, which are related to entry inhibitors developed by Bristol-Myers Squibb (BMS), have been synthesized with aliphatic chains located at the C7 position of the indole ring. These spacers contain an azido group suitable for the well-known Cu(I)-catalyzed (3+2)-cycloaddition or an activated triple bond for the nucleophilic addition of thiols under physiological conditions. Reaction with polyols (β-cyclodextrin and hyperbranched polyglycerol) decorated with complementary click partners has afforded polyol-BMS-like conjugates that are not cytotoxic (TZM.bl cells) and retain the activity against R5-HIV-1_NLAD8 isolates. Thus, potential vaginal microbicides based on entry inhibitors, which can be called of 4^th generation, are reported here for the first time.     

Molecular Genetics of Conjunctival Melanoma and Prognostic Value of TERT Promoter Mutation Analysis

The aim of this study was exploration of the genetic background of conjunctival melanoma (CM) and correlation with recurrent and metastatic disease. Twenty-eight CM from the Rotterdam Ocular Melanoma Study group were collected and DNA was isolated from the formalin-fixed paraffin embedded tissue. Targeted next-generation sequencing was performed using a panel covering GNAQ, GNA11, EIF1AX, BAP1, BRAF, NRAS, c-KIT, PTEN, SF3B1, and TERT genes. Recurrences and metastasis were present in eight (29%) and nine (32%) CM cases, respectively. TERT promoter mutations were most common (54%), but BRAF (46%), NRAS (21%), BAP1 (18%), PTEN (14%), c-KIT (7%), and SF3B1 (4%) mutations were also observed. No mutations in GNAQ, GNA11, and EIF1AX were found. None of the mutations was significantly associated with recurrent disease. Presence of a TERT promoter mutation was associated with metastatic disease (p-value = 0.008). Based on our molecular findings, CM comprises a separate entity within melanoma, although there are overlapping molecular features with uveal melanoma, such as the presence of BAP1 and SF3B1 mutations. This warrants careful interpretation of molecular data, in the light of clinical findings. About three quarter of CM contain drug-targetable mutations, and TERT promoter mutations are correlated to metastatic disease in CM.     

Real-time control of power systems using nodal prices

This article presents a novel control scheme for achieving optimal power balancing and congestion management in electrical power systems via nodal prices. We develop a dynamic controller that guarantees economically optimal steady-state operation while respecting all line flow constraints in steady-state. A benchmark example illustrates the effectiveness of the proposed control scheme.     

Estimating intramuscular fat content and fatty acid composition in live and post-mortem samples in pigs

Repeated muscle sampling (n = 732) on 216 pigs was performed to evaluate the effect of live sampling and sample size on intramuscular fat (IMF) content and fatty acid (FA) composition. The sampling scheme consisted of 1–3 biopsies of longissimus dorsi (LM), a small and a big post-mortem sample of LM, and a big post-mortem sample of gluteus medius (GM). IMF was determined by quantitative gas chromatography after direct transesterification. Data on LM were jointly analyzed using a mixed model on age with heterogeneous residual variances across sampling methods. Biopsies overestimated IMF and polyunsaturated FA content and underestimated monounsaturated FA content with decreasing sample size. Potential for bias and not sampling variance is the major limitation of using live samples for assessing changes in IMF with age. Small post-mortem samples of LM were as informative as big samples of GM for inferring IMF content but not IMF composition.     

On the prediction of delamination during deep-drawing of polymer coated metal sheet

This paper describes the development and application of a numerical model that can predict the delamination of the polymer coating from the steel substrate during deep-drawing. Experimentally characterized cohesive zones are used to describe the interface between the polymer and the steel and are capable of modelling delamination or a prior partial loss of adhesion during an axisymmetric deep-drawing simulation. A parameter is proposed that quantifies the interfacial integrity and is used to assess the influence of the tooling radii, the clearance between the punch and the die and the coating thickness on the interfacial integrity.     

Application of the Soil and Water Assessment Tool for six watersheds of Lake Erie: Model parameterization and calibration

The Soil and Water Assessment Tool (SWAT), a physically-based watershed-scale model, holds promise as a means to predict tributary sediment and nutrient loads to the Laurentian Great Lakes. In the present study, model performance is compared across six watersheds draining into Lake Erie to determine the applicability of SWAT to watersheds of differing characteristics. After initial model parameterization, the Huron, Raisin, Maumee, Sandusky, Cuyahoga, and Grand SWAT models were calibrated (1998-2001) and confirmed, or validated (2002-2005), individually for stream water discharge, sediment loads, and nutrient loads (total P, soluble reactive P, total N, and nitrate) based on available datasets. SWAT effectively predicted hydrology and sediments across a range of watershed characteristics. SWAT estimation of nutrient loads was weaker although still satisfactory at least two-thirds of the time across all nutrient parameters and watersheds. SWAT model performance was most satisfactory in agricultural and forested watersheds, and was less so in urbanized settings. Model performance was influenced by the availability of observational data with high sampling frequency and long duration for calibration and confirmation evaluation. In some instances, it appeared that parameter adjustments that improved calibration of hydrology negatively affected subsequent sediment and nutrient calibration, suggesting trade-offs in calibrating for hydrologic vs. water quality model performance. Despite these considerations, SWAT accurately predicted average stream discharge, sediment loads, and nutrient loads for the Raisin, Maumee, Sandusky, and Grand watersheds such that future use of these SWAT models for various scenario testing is reasonable and warranted.