Marker assisted selection for Varroa destructor resistance in New Zealand honey bees.

Varroa destructor is a honey bee (Apis mellifera) parasite identified as one of the leading causes of overwintering colony loss in New Zealand. It has been shown that a naturally occurring heritable trait, "Varroa Sensitive Hygiene" (VSH), confers an advantage to colonies by increasing behaviours that limit the survival and reproduction of Varroa mites. The SNP 9-9224292 is an adenine/guanine (A/G) polymorphism on chromosome 9 of Apis mellifera where the G allele was observed to be associated with VSH behaviour in North American honey bees. In this study, we sought to determine if selection for the G allele of SNP 9-9224292 could decrease Varroa mite infestation of New Zealand honey bee (Apis mellifera ligustica) colonies. We genotyped queens and tracked their colonies over summer before measuring Varroa levels at the point of autumn Varroa treatment. The mean Varroa population level in colonies headed by queens that carry two copies of VSH associated G allele of SNP 9-9224292 was 28.5% (P<0.05) lower compared with colonies headed by queens with two copies of non-VSH associated A alleles. Although a significant reduction in mite infestation was achieved in treatment colonies, conventional Varroa treatment was still required for adequate Varroa control. Considering the open mating of queens used and a lack of drift control in this study, this VSH SNP shows promise for marker assisted selection of New Zealand honey bees when aiming for innate Varroa control traits.

Periodicity in Structure, Bonding, and Reactivity for p-Block Complexes of a Geometry Constraining Triamide Ligand.

The use of pincer ligands to access non-VSEPR geometries at main-group centers is an emerging strategy for eliciting new stoichiometric and catalytic reactivity. As part of this effort, several different tridentate trianionic substituents have to date been employed at a range of different central elements, providing a patchwork dataset that precludes rigorous structure-function correlation. An analysis of periodic trends in structure (solid, solution, and computation), bonding, and reactivity based on systematic variation of the central element (P, As, Sb, or Bi) with retention of a single tridentate triamide substituent is reported herein. In this homologous series, the central element can adopt either a bent or planar geometry. The tendency to adopt planar geometries increases descending the group with the phosphorus triamide (1) and its arsenic congener (2) exhibiting bent conformations, and the antimony (3) and bismuth (4) analogues exhibiting a predominantly planar structure in solution. This trend has been rationalized using an energy decomposition analysis. A rare phase-dependent dynamic covalent dimerization was observed for 3 and the associated thermodynamic parameters were established quantitatively. Planar geometries were found to engender lower LUMO energies and smaller band gaps than bent ones, resulting in different reactivity patterns. These results provide a benchmark dataset to guide further research in this rapidly emerging area.

Simultaneous Ligand and Receptor Tracking through NMR Spectroscopy Enabled by Distinct 19F Labels.

To probe ligand-receptor binding at the atomic-level, a frequent approach involves multidimensional nuclear magnetic resonance (NMR) spectroscopy experiments relying on 13C- and/or 15N-enrichment alongside 1H. Alternatively, the lack of fluorine in biomolecules may be exploited through specific incorporation of 19F nuclei into a sample. The 19F nucleus is highly sensitive to environmental changes and allows for one-dimensional NMR spectroscopic study, with perturbation to chemical shift and spin dynamics diagnostic of structural change, ligand binding, and modified conformational sampling. This was applied to the apelinergic system, which comprises a rhodopsin-like G protein-coupled receptor (the apelin receptor (AR)/APJ) and two families of cognate ligands, the apelin and apela (ELABELA/toddler) peptides. Specifically, AR fragments consisting of either the N-terminal tail and first transmembrane (TM) α-helix (AR55) or the first three transmembrane α-helices (TM1-3) were prepared with biosynthetic fluorotryptophan incorporation. Interactions of each AR fragment with a high-affinity, 2,4,5-trifluorophenylalanine labeled apelin analogue were compared by 19F NMR. Distinct ranges of 19F chemical shifts for ligand and receptor provide unambiguous tracking of both species, with distinct binding behaviour observed for each AR fragment implying that AR55 is not sufficient to recapitulate the physiological binding event. Site-specific perturbation was also apparent for the apelin analogue as a function of substitution site, indicating an orientational binding preference. As a whole, this strategy of distinctive 19F labelling for ligand and receptor provides a relatively fast (i.e., employing 1D NMR experiments) and highly sensitive method to simultaneously and definitively track binding in both species.

New ¹H NMR-Based Technique To Determine Epoxide Concentrations in Oxidized Oil.

A new method to determine epoxide concentrations in oxidized oils was developed and validated using (1)H NMR. Epoxides derived from lipid oxidation gave signals between 2.90 and 3.24 ppm, well separated from the signals of other lipid oxidation products. To calibrate, soybean oils with a range of epoxide concentrations were synthesized and analyzed using (1)H NMR by taking the sn-1,3 glycerol protons (4.18, 4.33 ppm) as internal references. The (1)H NMR signals were compared to the epoxide content determined by titration with hydrogen bromide (HBr)-acetic acid solution. As expected, the signal response increased with concentration linearly (R(2) = 99.96%), and validation of the method gave results comparable to those of the HBr method. A study of the oxidative stability of soybean oil was performed by applying this method to monitor epoxides during thermal lipid oxidation. The epoxide content increased over time and showed a different trend compared to peroxide value (PV). A phenomenological model was suggested to model epoxides derived from lipid oxidation.

Preparation of a diphosphine with persistent phosphinyl radical character in solution: characterization, reactivity with O2, S8, Se, Te, and P4, and electronic structure calculations.

A new, easily synthesized diphosphine based on a heterocyclic 1,3,2-diazaphospholidine framework has been prepared. Due to the large, sterically encumbering Dipp groups (Dipp = 2,6-diisopropylphenyl) on the heterocyclic ring, the diphosphine undergoes homolytic cleavage of the P-P bond in solution to form two phosphinyl radicals. The diphosphine has been reacted with O(2), S(8), Se, Te, and P(4), giving products that involve insertion of elements between the P-P bond to yield the related phosphinic acid anhydride, sulfide/disulfide, selenide, telluride, and a butterfly-type perphospha-bicyclobutadiene structure with a trans,trans-geometry. All molecules have been characterized by multinuclear NMR spectroscopy, elemental analysis, and single-crystal X-ray crystallography. Variable-temperature EPR spectroscopy was utilized to study the nature of the phosphinyl radical in solution. Electronic structure calculations were performed on a number of systems from the parent diphosphine [H(2)P](2) to amino-substituted [(H(2)N)(2)P](2) and cyclic amino-substituted [(H(2)C)(2)(NH)(2)P](2); then, bulky substituents (Ph or Dipp) were attached to the cyclic amino systems. Calculations on the isolated diphosphine at the B3LYP/6-31+G* level show that the homolytic cleavage of the P-P bond to form two phosphinyl radicals is favored over the diphosphine by ~11 kJ/mol. Furthermore, there is a significant amount of relaxation energy stored in the ligands (52.3 kJ/mol), providing a major driving force behind the homolytic cleavage of the central P-P bond.

Comprehensive chemical characterization of complexes involving lead-amino acid interactions.

Complexes of lead with L-phenylalanine, L-isoleucine, L-valine, or L-arginine have been isolated from reaction mixtures containing lead nitrate and the respective amino acid in acidic aqueous solution. The compounds have been comprehensively characterized using X-ray crystallography, solid state NMR spectroscopy and solution state NMR spectroscopy, IR and Raman spectroscopies, and electrospray ionization mass-spectrometry. The solid state structures of lead-phenylalanine, lead-valine, and lead-valine-isoleucine complexes show a lead center coordinated by two amino acid ligands, while the lead-arginine complex is a cluster involving two lead centers and three arginine molecules. The structural, spectroscopic, and spectrometric characterization of the complexes provides a basis to establish a fundamental understanding of heavy metal-amino acid interactions.

31P NMR studies demonstrating the assembly of catena-phosphorus frameworks from chlorophosphinochlorophosphonium cations.

New examples of chlorophosphinochlorophosphonium (4) and chlorophosphinodichlorophosphonium (5) cations have been prepared and spectroscopically characterized. These bifunctional phosphinophosphonium cations offer a new approach to the development of phosphinophosphonium frameworks using reductive coupling reactions and have been exploited as synthons to assemble larger catena-phosphorus cations. The reactions of 4 and 5 with stibine reducing agents have been studied using (31)P NMR spectroscopy and have been shown to produce a variety of new and known frameworks in a facile manner, depending on the reducing agent selection and the stoichiometry of the reaction. New derivatives of frameworks containing three, four, and five phosphorus atoms have been identified by (31)P NMR spectroscopy. Crystals of the [GaCl(4)](-) salt of the five-membered dication 11((i)Pr) have been isolated from the reaction of [4((i)Pr)][GaCl(4)] with SbBu(3), and the solid state structural features and solution state dynamics are comprehensively described in the context of (31)P{(1)H} NMR spectroscopic observations.

2-Propynyl 2,3,4,6-tetra-O-acetyl-α-D-mannopyranoside.

The 2-propynyl group in the title compound, C(17)H(22)O(10), adopts an exoanomeric conformation, with the acetylenic group gauche with respect to position C1. Comparison of (13)C NMR chemical shifts from solution and the solid state suggest that the acetylenic group also adopts a conformation anti to C1 in solution. The pyranose ring adopts a (4)C(1) conformation. Of the three secondary O-acetyl groups, that on position O4, flanked by two equatorial groups, adopts a syn conformation, in agreement with recent generalizations [González-Outeiriño, Nasser & Anderson (2005). J. Org. Chem. 70, 2486-2493]. The acetyl group on position O3 adopts a gauche conformation, also in agreement with the recent generalizations, but that on position O2 adopts a syn conformation, not in agreement with the recent generalizations.

2,3-Diphosphino-1,4-diphosphonium ions.

Salts of the first crystallographically characterized chlorophosphinophosphonium ions have been prepared, and their reaction with Ph3P results in reductive coupling of the chlorophosphine centers to give the first acyclic 2,3-diphosphino-1,4-diphosphonium ions, representing a key framework in the development of catena-phosphorus chemistry. These new salts of general formula [R3P-PR'-PR'-PR3][OTf]2 are also obtained in a one-pot diastereoselective reaction of a dichlorophosphine, a tertiary phosphine, and trimethylsilyltrifluoromethanesulfonate. The structural and spectroscopic features of the new dications complement those of the known diphosphonium and 2-phosphino-1,3-diphosphonium dications. Quantitative ligand exchange reactions are observed when derivatives of [Ph3P-PR'-PR'-PPh3][OTf]2 are combined with Me3P, demonstrating the coordinative nature of the phosphine-phosphonium P-P bonds and implicating a bonding model involving the diphosphenium dication acceptor. The observed solid state structures have been interpreted in the context of computational studies.

Glycosidase inhibition by macrolide antibiotics elucidated by STD-NMR spectroscopy.

A glycosynthase approach was attempted to glycodiversify macrolide antibiotics, using DesR, a family-3 retaining beta-glucosidase involved in the self-resistance mechanism of methymycin production. STD-NMR was used to probe enzyme-substrate interactions. Analysis of competitive STD-NMR experiments between erythromycin A and a chromogenic substrate (pNP-beta-d-glucose) with the hydrolytically inactive nucleophile mutants led us to discover a family of unprecedented glycosidase inhibitors. Analysis of kinetic data with wild-type DesR determined that erythromycin is a competitive inhibitor of the glucosidase (IC50 = 2.8 +/- 0.3 microM and Ki = 2 +/- 0.2 microM) with respect to the hydrolysis of pNP-beta-d-glucose. Comparable inhibitory data was obtained for clarithromycin; however, the inhibitory effect of azithromycin was weak and no significant inhibition was observed with methymycin or d-desosamine. This report documents significant inhibition of glycosidases by macrolide antibiotics and provides insight into the design of novel glycosidase inhibitors based on the macrolactone ring of macrolide antibiotics.

Synthesis and characterization of elusive cyclo-di- and -tri-phosphino-1,3-diphosphonium salts: fundamental frameworks in catena-organophosphorus chemistry.

Reaction of a cyclophosphinophosphonium cation with neat MeOTf represents a general and high-yield synthetic approach to dications enabling the isolation of the first derivatives of 2,4,5-triphosphino-1,3-diphosphonium as bis-triflate salts.

Cyclotetraphosphinophosphonium ions: synthesis, structures, and pseudorotation.

The first derivatives of catenated cyclotetraphosphinophosphonium cations, [(PhP)4PPhMe]+ (8a), [(MeP)4PMe2]+ (8b), [(CyP)4PPh2]+ (8d), [(CyP)4PMe2]+ (8e), [(PhP)4PPh2]+ (8f), [(PhP)4PMe2]+ (8g), are synthesized as trifluoromethanesulfonate (triflate, OSO2CF3-) salts through the reaction of cyclopentaphosphines (PhP)5 (4a) or (MeP)5 (4b) with methyl triflate (MeOTf) or by a net phosphenium ion [PR2+, R = Ph, Me; from R2PCl and trimethylsilyltriflate (Me3SiOTf)] insertion into the P-P bond of either cyclotetraphosphine (CyP)4 (3c) or cyclopentaphosphines (PhP)5 (4a) or (MeP)5 (4b). Although more conveniently prepared from 4a, compound 8a[OTf] can also be formed from (PhP)4 (3a) and MeOTf, and derivatives 8f[OTf] and 8g[OTf] are also accessible through reactions of 3a and R2PCl/Me3SiOTf with R = Ph or Me, respectively. A tetrachlorogallate salt of [(PhP)4PPhtBu]+ (8c) has been synthesized by alkylation of 4a with tBuCl/GaCl3. 31P[1H] NMR parameters for all derivatives of 8 have been determined by iterative simulation of experimental data. Derivatives 8a[OTf], 8b[OTf], 8c[GaCl4], 8e[OTf], 8f[OTf], and 8g[OTf] and have been characterized by X-ray crystallography, showing the most favorable all-trans configuration of substituents for the phosphine centers, thus minimizing steric interactions. Each derivative adopts a unique envelope or twist conformation of C1 symmetry. The effective C2 symmetry observed for 8b, d, e, f, and g in solution, signified by their 31P[1H] NMR AA'BB'X spin systems, implies a rapid conformational exchange for derivatives of 8. The core frameworks of the cations in the solid state are viewed as snapshots of different conformational isomers within the solution-phase pseudorotation process.

Monocyclic di- and triphosphinophosphonium cations: new foundational frameworks for catena-phosphorus chemistry.

The synthesis and characterization for trifluoromethanesulfonate (triflate) salts of the first definitive examples of cyclotriphosphinophosphonium and cyclodiphosphinophosphonium cations are described, representing new prototypical frameworks in the rational and systematic development of catena-phosphorus chemistry. Addition of methyl triflate (MeOTf) or triflic acid (HOTf) to cyclotetraphosphines (tBuP)4 (1a) or (CyP)4 (1b) gives [(tBuP)3PtBuMe][OTf] (2a[OTf]), [(CyP)3PCyMe][OTf] (2b[OTf]), [(tBuP)3PtBuH][OTf] (3a[OTf]), and [(CyP)3PCyH][OTf] (3b[OTf]), respectively. Cyclotriphosphine (tBuP)3 (4a) reacts with HOTF or Me2PCl/Me3SiOTf to give the ring expanded cations 3a[OTf] and [(tBuP)3PMe2][OTf] (5[OTf]), respectively, but reactions with MeOTf and HCl give cyclic diphosphinophosphonium cation [(tBuP)2PtBuMe][OTf] (6a[OTf]) and ring-opened triphosphine HtBuP-PtBu-PtBuCl (7), respectively. The analogous diphosphinophosphonium cation [(CyP)2PCyMe][OTf] (6b[OTf]) is formed along with 2b[OTf] in reactions of MeOTf with (CyP)3 (4b). Compounds 2a[OTf], 2b[OTf], 3a[OTf], 5[OTf], and 6a[OTf] have been crystallographically characterized. 1H NMR spectra of 2a[OTf], 2b[OTf], 5[OTf], and 6a[OTf] demonstrate that 3JPH coupling is only observed for methyl protons if they are in a cis orientation to the lone pairs on the adjacent phosphine sites.

Immunostimulatory polysaccharides from Chlorella pyrenoidosa. A new galactofuranan. measurement of molecular weight and molecular weight dispersion by DOSY NMR.

Fractionation of the hot water extract of Chlorella pyrenoidosa was performed using a combination of ethanol precipitation, size exclusion chromatography, and anion exchange chromatography. One fraction contained a new polysaccharide, and this compound was shown to be a 1-->2-linked beta-d-galactofuranan from its 1D and 2D (1)H and (13)C NMR spectra, with a molecular weight of 15 kDa from DOSY NMR measurements. A number of other fractions were shown to have the same repeating unit as the previously identified arabinogalactan. However, arabinogalactans from different fractions were shown by DOSY NMR to have different molecular weights, which ranged from 27 to 1020 kDa. Agreement with molecular weights measured for some of these fractions by SEC-MALS was very good, further confirming the relationship established by Viel et al. between molecular weights of neutral polysaccharides and self-diffusion coefficients. The smaller molecular weight polysaccharides, the galactofuranan and the 27 and 50 kDa arabinogalactans, were shown to be close to monodisperse by analysis of the distributions of the self-diffusion coefficients for the polymers. The larger arabinogalactans had considerable variation in their molecular weights (188 +/- 109 kDa and 1020 +/- 370 kDa). Only the two larger arabinogalactans showed immunostimulatory activity.

Acyclic catena-diphosphinodiphosphonium dications [R3P-PR'-PR'-PR3]2+ or bisphosphine-diphosphenium complexes [R3PPR'-PR'PR3]2+: synthesis by reductive P-P coupling of [R3P-PR'Cl]+ and phosphine ligand exchange.

The acyclic tetraphosphorus dication [Ph3P-PPh-PPh-PPh3]2+ has been formed by the reductive coupling of [Ph3P-PPhCl]+, providing a new synthetic method for the systematic development of catena-phosphorus cations. Ligand exchange (Ph3P for Me3P) gives [Me3P-PPh-PPh-PMe3]2+, implicating these dications as bisphosphine-diphosphenium complexes.

Isolation, characterization and structural determination of a unique type of arabinogalactan from an immunostimulatory extract of Chlorella pyrenoidosa.

An arabinogalactan was isolated from a hot water extract of freeze-dried cells of the green microalga, Chlorella pyrenoidosa. This hot water extract is a proprietary immunomodulator, with the trademark Respondintrade mark (ONC-107). The arabinogalactan was recovered from the ethanol-soluble fraction of the supernatant resulting from a process that involved controlled ethanol precipitation followed by size exclusion chromatography on Sephadex G-100, then Cetavlon precipitation. Sugar analyses, GC-MS data for (S)-2-octyl glycosides, and 1D and 2D NMR experiments established unambiguously that the repeating unit was -->2)-alpha-L-Araf-(1-->3)-[alpha-L-Araf-(1-->4)]-beta-D-Galp-(1-->. This structure does not fit into any of the known classes of arabinogalactans. SEC/MALS experiments gave a molecular mass for the arabinogalactan isolated as 47 +/- 4 kDa but the original structure was probably larger.

Hypervalent, low-coordinate phosphorus(III) centers in complexes of the phosphadiazonium cation with chelate ligands.

Trifluoromethylsulfonyloxy-(2,4,6-tri-tert-butylphenylimino)phosphine, Mes*NPOTf (Mes = 2,4,6-tri-tert-butylphenyl, OTf = trifluoromethanesulfonate, triflate) reacts quantitatively with the multifunctional ligands 2,2'-bipyridine (2,2'-BIPY), N,N,N',N'-tetramethylethylenediamine (TMEDA), 1,2-bis(diethylphosphino)ethane (DEPE), 1,2-bis(diphenylphosphino)ethane (DIPHOS), and N,N,N',N' ',N' '-pentamethyldiethylenetriamine (PMDETA) to give the Lewis acid-base complexes [Mes*NP(2,2'-BIPY)][OTf], [Mes*NP(TMEDA)][OTf], [Mes*NP(DIPHOS)][OTf], [Mes*NP(DEPE)][OTf], and [Mes*NP(PMDETA)][OTf], respectively. Single-crystal X-ray diffraction studies indicate that the closest contact of the ligand donor atoms occurs at phosphorus in all cases, affecting significant displacement of the OTf anion. The resulting cations [Mes*NP(L)]+ are best described as complexes of a neutral chelating ligand on a phosphadiazonium Lewis acceptor, and highlight the potential for electron-rich centers to behave as Lewis acids despite the presence of a lone pair of electrons at the acceptor site. More importantly, the new complexes represent rare examples of systems containing hypervalent, low-coordinate phosphorus(III) centers.

Transformations between monomeric, dimeric, and trimeric phosphazanes: oligomerizing NP analogues of olefins.

Isolation and characterization of a crystal mixture of iminophosphine 1 and diphosphazane 2 (R = Mes*, X = OTf) is enabled by the steric interactions between bulky substituents implicating monomer/dimer 1/2 equilibria and the conclusion is supported by the observation of a ring-expansion reaction to give a triphosphazane 3 (R = Dipp, X = Cl).