Inhibitors of an essential mycobacterial cell wall lipase (Rv3802c) as tuberculosis drug leads

The first targeted inhibitors of an essential M. tuberculosis cell wall lipase, Rv3802c, are described. Lead compounds exhibited nanomolar inhibition of the enzyme, and encouraging antibacterial activity against M. tuberculosis in vitro, supporting Rv3802c as a novel TB drug target.     

Comparative analysis of shape memory-based self-healing coatings

Self-healing materials exhibit the ability to repair and to recover their functionality upon damage. Here, we report on an investigation into preparation and characterization of shape memory assisted self-healing coatings. We built on past work in which poly(ε-caprolactone) electrospun fibers were infiltrated with a shape memory epoxy matrix and delve into fabricating and characterizing a coating with the same materials, but employing a blending approach, polymerization induced phase separation. After applying controlled damage, the ability of both coatings to self-heal upon heating was investigated. In both methods, coatings showed excellent thermally induced crack closure and protection against corrosion, with the blend approach being more suitable for large-scale applications given its process simplicity. Two different approaches to the preparation of shape memory-based self-healing coatings were compared for their ability to heal structurally and functionally by heating. These two approaches, electrospinning versus polymerization-induced phase separation were found to feature comparable and quite complete healing, with the latter system offering the advantage of facile processing. © 2016 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2016 , 54, 1415–1426     

Determination,by inductively coupled plasma mass spectrometry,of changes in cellular metal content resulting from herpes simplex virus-1 (HSV-1) infection

Metals and metal-containing compounds are known to play important roles in many biological processes, including metabolic and detoxification pathways and the formation and function of proteins. Like all organisms, viruses are expected to contain different metals. These metals, either by themselves or in the form of metalloproteins, may be involved in the virus’s ability to infect healthy cells and replicate within them. Identification and speciation of metals in control cells and in cells affected by a virus could be helpful in elucidating infection and replication mechanisms; these might, in turn, be vital to the development of more effective treatments. There has, however, been no extensive investigation of the metals specific viruses contain or affect. The objective of this study was to investigate changes in cellular metal content resulting from herpes simplex virus 1 (HSV-1) infection. Inductively coupled plasma mass spectrometry was used to identify differences between metal concentrations in uninfected and HSV-1-infected mammalian cells. Although it can be assumed that decreases in metal content are a result of cellular response to the virus, increases can be attributed either to cellular response or to the HSV-1 virus itself. Microwave digestion and flow injection methods suitable for small sample volumes were used, and the effects of different virus inactivation procedures were explored. This work is the first step in the identification of metals pertinent to HSV-1 infection and lays the foundation for future studies concentrating on characterization of these metal-associated or containing molecules.     

Rates of proton transfer to Fe-S-based clusters: comparison of clusters containing {MFe(mu(2)-S)(2)}n+ and {MFe(3)(mu(3)-S)(4)}n+ (M = Fe, Mo, or W) cores

The rates of proton transfer from [pyrH]+ (pyr = pyrrolidine) to the binuclear complexes [Fe2S2Cl4]2- and [S2MS2FeCl2]2- (M = Mo or W) are reported. The reactions were studied using stopped-flow spectrophotometry, and the rate constants for proton transfer were determined from analysis of the kinetics of the substitution reactions of these clusters with the nucleophiles Br- or PhS- in the presence of [pyrH]+. In general, Br- is a poor nucleophile for these clusters, and proton transfer occurs before Br- binds, allowing direct measure of the rate of proton transfer from [pyrH]+ to the cluster. In contrast, PhS- is a better nucleophile, and a pathway in which PhS- binds preferentially to the cluster prior to proton transfer from [pyrH]+ usually operates. For the reaction of [Fe2S2Cl4]2- with PhS- in the presence of [pyrH]+ both pathways are observed. Comparison of the results presented in this paper with analogous studies reported earlier on cuboidal Fe-S-based clusters allows discussion of the factors which affect the rates of proton transfer in synthetic clusters including the nuclearity of the cluster core, the metal composition, and the nature of the terminal ligands. The possible relevance of these findings to the protonation sites of natural Fe-S-based clusters, including FeMo-cofactor from nitrogenase, are presented.     

Optimization of DNA-tagged liposomes for use in microtiter plate analyses

Dye-encapsulating unilamellar DNA oligonucleotide-tagged liposomes were prepared and characterized for use as signal-enhancing reagents in a microtiter plate sandwich-hybridization analyses of single-stranded RNA or DNA sequences. The liposomes were synthesized using the reversed-phase evaporation method and tagged with DNA oligonucleotides by adding cholesteryl-modified DNA reporter probes to the initial lipid mixture. Liposomes were prepared using probe coverages of 0.0013–0.103 mol% of the total lipid input, several hydrophobic and poly(ethylene glycol)-based spacers between the cholesteryl anchor and the probe, and liposome diameters ranging from 200 nm to 335 nm. Their signal enhancement functionality was compared by using them in microtiter plate sandwich-hybridization assays for the detection of single-stranded DNA sequences. In these assays, an optimal reporter probe concentration of 0.103 mol%, a liposome diameter of 274 nm, and a phospholipid concentration of 0.3 mM were found. The length between the cholesteryl anchor and the probe was optimal when a spacer composed of TEG+(CH₂O)₃ was used. Under optimal conditions, a detection limit of 0.5 nM for a truncated synthetic DNA sequence was found with a coefficient of variation of 4.4%. A 500-fold lower limit of detection using fluorescence was found using lysed dye-encapsulating liposomes versus a single fluorescein-labeled probe. Finally, when this method was applied to the detection of atxA RNA extracted from E.coli SG12036-pIu121 and amplified using NASBA, a minimum extracted concentration of RNA of 1.1×10⁻⁷ μg/μL was found.     

Optimization of DNA-tagged dye-encapsulating liposomes for lateral-flow assays based on sandwich hybridization

A novel protocol for the synthesis of dye-encapsulating liposomes tagged with DNA oligonucleotides at their outer surface was developed. These liposomes were optimized for use as signal enhancement agents in lateral-flow sandwich-hybridization assays for the detection of single-stranded RNA and DNA sequences. Liposomes were synthesized using the reverse-phase evaporation method and tagged with oligonucleotides by adding cholesteryl-modified DNA probes to the initial lipid mixture. This resulted in a greatly simplified protocol that provided excellent control of the probe coverage on the liposomes and cut the preparation time from 16 hours to just 6 hours. Liposomes were prepared using probe concentrations ranging from 0.00077 to 0.152 mol% of the total lipid, several hydrophobic and polyethylene glycol-based spacers between the cholesteryl anchor and the probe, and liposome diameters ranging from 208 nm to 365 nm. The liposomes were characterized by dynamic light scattering, visible spectroscopy, and fluorescence spectroscopy. Their signal enhancement functionality was compared by using them in lateral-flow optical biosensors for the detection of single-stranded DNA sequences. In these assays, an optimal reporter probe concentration of 0.013 mol%, liposome diameter of 315 nm, and liposome optical density of 0.4–0.6 at 532 nm were found. The spacer length between the cholesteryl anchor and the probe showed no significant effect on the signals in the lateral-flow assays. The results presented here provide important data for the general use of liposomes as labels in analytical assays, with specific emphasis on nucleic acid detection via lateral flow assays.     

Heterogeneous catalysts for cyclic carbonate synthesis from carbon dioxide and epoxides

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The boron-mediated ketone-ketone aldol reaction

The first examples of the directed, boron-mediated aldol reaction between different ketones are presented. Transformation of a variety of ketones to their corresponding boron enolates with Chx₂BCl/Et₃N, followed by reaction with acceptor ketones in diethyl ether, and oxidation of the resultant boron aldolate (H₂O₂, MeOH/pH 7 buffer), provided the aldol addition products. The reaction was most facile when cyclic ketones were used, with the highest yields obtained for the reaction of boron enolates with cyclohexanone as the acceptor.     

Three-component one-pot total syntheses of glyantrypine, fumiquinazoline F, and fiscalin B promoted by microwave irradiation

A microwave-promoted three-component one-pot reaction has been developed to provide access to the core pyrazino[2,1-b]quinazoline-3,6-dione (1) scaffold, which is common to several families of alkaloids with significant biological activities. By adapting this synthetic strategy through the use of selected Boc-amino acids and amino acid esters, we have accomplished highly efficient and concise total syntheses of glyantrypine (2), fumiquinazoline F (3), and fiscalin B (5), achieving overall yields of 55, 39, and 20%, respectively.