An assignment to internal soliton vibrations of laser-raman lines from living cells

The internal oscillations of an alpha-helix soliton (the Davydov soliton) are studied numerically and compared with recently measured laser-Raman spectra on metabolically active E. coli. Agreement between calculated and measured spectra strongly supports the idea that the soliton is a possible mechanism for transport and storage of biological energy in proteins.     

Novel sesquiterpenoids from the fermentation of Xylaria persicaria are selective ligands for the NPY Y5 receptor

Neuropeptide Y (NPY) is a polypeptide found in the peripheral and central nervous system and is involved in the regulation of feeding. Antagonists of NPY receptor activation could therefore have potential for development as antiobesity drugs. Fermentation of an isolate of Xylaria persicaria yielded two novel eremophilane sesquiterpenoids xylarenals A (1) and B (2). These compounds are selective for the NPY Y5 receptor but have only modest affinity. The isolation, structure elucidation, and biological activities of these compounds are described.     

Single-laboratory validation of a modified liquid chromatographic method with UV detection for determination of trenbolone residues in bovine liver and muscle

Trenbolone acetate is a synthetic testosterone analog registered for use in a number of countries as a growth-promoting hormone, applied as an implant in the ears of feedlot cattle. The method is intended for the detection and quantitation of trace amounts of alpha- and beta-trenbolone in bovine tissues (muscle, liver) by liquid chromatography (LC) with UV detection and eliminates the use of the structural analog, 19-nortestosterone, as an internal standard. Trenbolone residues are extracted from tissues that have been homogenized in sodium acetate with a 3-phase liquid-liquid extraction by adding a mixture of water-acetonitrile-dichloromethanehexane, with trenbolone residues preferentially partitioned into the middle acetonitrile layer. The extract is passed through solid-phase extraction cartridges (both C18 and silica gel) using, respectively, methanol-water and acetone-toluene as eluents. Reversed-phase high-performance LC separation is performed, an octadecyl-bonded column with methanol-acetonitrile-water used as mobile phase for sample analysis. The limit of detection is 0.2 ng/g in muscle tissue and 0.6 ng/g in liver tissue, with coefficients of variation of 3.5-12.1% for alpha- and beta-trenbolone at concentrations from 0.2 to 4.0 ng/g fortified in muscle and 3.3-26.0% from liver fortified at 0.6-10.0 ng/g. Absolute recoveries of 40-130% were observed, but the use of fortified matrix curves eliminated recovery correction. Critical control points were identified in a pH adjustment step and an evaporation step during method validation, which included ruggedness testing. Analysis of incurred tissues (bovine liver and muscle) stored at -20 degrees C for over 25 weeks did not identify any significant loss of residues.     

Determination of clopidol residues in chicken tissues by liquid chromatography: collaborative study

Eighteen laboratories participated in a collaborative study on the determination of clopidol residues in chicken muscle tissues by liquid chromatography. Of these, results from 16 laboratories which rigorously followed the method were subjected to statistical analysis. The method performance was assessed by all participants using 14 samples of chicken muscle fortified at concentrations ranging from 0.1 to 5.0 mg/kg. In addition, 9 participants each reported results for 6 clopidol-incurred samples in chicken muscle. Test portions were extracted with acetonitrile, and the extracts were purified with alumina and anion exchange resin solid-phase extraction cartridges in sequence. Clopidol was separated by reversed-phase liquid chromatography and quantified at 270 nm. Average recoveries ranged from 81.8 to 85.4%, reproducibility relative standard deviation (RSDR) ranged from 11.9 to 22.6%, and repeatability relative standard deviation (RSDr) ranged from 9.9 to 15.1%. For clopidol-incurred samples at concentrations of 0.100-0.687 mg/kg, the mean determination value range was 0.099-0.659 mg/kg; RSDR was 12.6-19.8%, RSDr was 3.1-8.5%; and HORRAT values were 0.7-1.1. The accuracy and precision of the method are in conformity with the requirements specified by AOAC INTERNATIONAL. The method was adopted Official First Action in April 2003.     

Validation of screening method for residues of diethylstilbestrol, dienestrol, hexestrol, and zeranol in bovine urine using immunoaffinity chromatography and gas chromatography/mass spectrometry

A method was developed, using commercially available immunoaffinity chromatography cleanup cartridges, followed by detection by gas chromatography/mass spectrometry, to screen for residues of the hormone growth promotants diethylstilbestrol, dienestrol, hexestrol, and zeranol in bovine urine. The single-laboratory, in-house validation included assessment of recoveries, repeatability, linearity of response, detection capability, and specificity (cross-reactivity) with a suite of antibiotics and other hormonal growth promotants. The method was validated for screening at a target concentration of 2.0 microg/L in urine. The detection capabilities for the analytes were diethylstilbestrol, 0.24; dienestrol, 0.15; hexestrol, 0.84; and zeranol, 0.28 microg/L.     

Determination of beta-agonists in liver and retina by liquid chromatography-tandem mass spectrometry

A liquid chromatography-tandem mass spectrometry (LC/MS/MS) method for the determination of bromobuterol, cimaterol, clenbuterol, clenpenterol, hydroxymethylclenbuterol, isoxsuprine, mabuterol, ractopamine, ritrodrine, salbutamol, terbutaline, and tulobuterol residues in bovine liver and retina is reported. This procedure uses enzymatic digestion, liquid-liquid extraction, and cleanup on Oasis HLB solid-phase extraction cartridges, followed by determination of the residues by LC-tandem quadrupole MS using atmospheric pressure chemical ionization in the positive ion mode. Overall average recoveries ranged from 23 to 76% for liver and 34 to 77% for retina. The mean values for samples fortified at levels between 0.5-2.0 microg/kg (liver) and 5-20 microg/kg (retina) agreed within 98-118% of the spiked levels, with coefficients of variation ranging from 6 to 20%. The decision limits, CCalpha, ranged from 0.1 to 0.3 microg/kg for liver, 1-3 microg/kg for retina, and detection capabilities, CCbeta, from 0.2-0.5 microg/kg for liver and 2-5 microg/kg for retina.     

Polymethacrylate Networks as Substrates for Cell Culture

Summary: methacrylate networks have a long history of applications in medical technology and much is known of their non-fouling properties. However, in recent times it has become clear that the swollen nature of these materials may provide some advantages if they are used as scaffolds in tissue engineering. In general however these hydrogels are resistant to protein adsorption and human cells do not easily adhere. In this work we provide an overview of several strategies that are designed to improve the cell-adhesive properties of hydrogels while maintaining their useful properties, mainly ease of diffusion of nutrients and growth factors. We describe our early attempts at modifying hydrogels based on 2,3-propandiol -1-methacrylate, with either hydrophobic units or acid groups. Modification with lauryl methacrylate produced an improvement but acid modification failed to provide surfaces that were conducive to cell culture. Much better scaffolds were prepared by amination of epoxy functional 2,3-propandiol-1-methacrylate networks. Optimized materials in this class were shown to be good substrates for the co-culture of bovine keratocytes with human corneal epithelial cells. We also describe the synthesis and biological properties of methacrylate conetworks, which phase separate during synthesis to give porous amphiphilic materials. Optimization of these materials produces materials that perform as well as tissue culture plastic so that confluent sheets of human dermal fibroblasts can be produced using standard culture techniques.     

An H-Substituted Rhodium Silylene

Divergent reactivity of organometallic rhodium(I) complexes, which led to the isolation of neutral rhodium silylenes, is described. Addition of PhRSiH₂ (R=H, Ph) to the rhodium cyclooctene complex (^iPrNNN)Rh(COE) (1-COE; ^iPrNNN=2,5-[iPr₂P=N(4-iPrC₆H₄)]₂N(C₆H₂)⁻, COE=cyclooctene) resulted in the oxidative addition of an Si−H bond, providing rhodium(III) silyl hydride complexes (^iPrNNN)Rh(H)SiHRPh (R=H, 2 -SiH₂Ph; Ph, 2 -SiHPh₂). When the carbonyl complex (^iPrNNN)Rh(CO) ( 1 -CO) was treated with hydrosilanes, base-stabilized rhodium(I) silylenes κ²-N,N-(^iPrNNN)(CO)Rh=SiRPh (R=H, 3 -SiHPh; Ph, 3 -SiPh₂) were isolated and characterized using multinuclear NMR spectroscopy and X-ray crystallography. Both silylene species feature short Rh−Si bonds [2.262(1) Å, 3 -SiHPh; 2.2702(7) Å, 3 -SiPh₂] that agree well with the DFT-computed structures. The overall reaction led to a change in the ^iPrNNN ligand bonding mode (κ³→κ²) and loss of H₂ from PhSiRH₂, as corroborated by deuterium labelling experiments.     

Synthesis and Reactivity of Organometallic Intermediates Relevant to Cobalt‐Catalyzed Hydroformylation

Intermediates relevant to cobalt‐catalyzed alkene hydroformylation have been isolated and evaluated in fundamental organometallic transformations relevant to aldehyde formation. The 18‐electron (R,R)‐(^iPrDuPhos)Co(CO)₂H has been structurally characterized, and it promotes exclusive hydrogenation of styrene in the presence of 50 bar of H₂/CO gas (1:1) at 100 °C. Deuterium‐labeling studies established reversible 2,1‐insertion of styrene into the Co?D bond of (R,R)‐(^iPrDuPhos)Co(CO)₂D. Whereas rapid β‐hydrogen elimination from cobalt alkyls occurred under an N₂ atmosphere, alkylation of (R,R)‐(^iPrDuPhos)Co(CO)₂Cl in the presence of CO enabled the interception of (R,R)‐(^iPrDuPhos)Co(CO)₂C(O)CH₂CH₂Ph, which upon hydrogenolysis under 4 atm H₂ produced the corresponding aldehyde and cobalt hydride, demonstrating the feasibility of elementary steps in hydroformylation. Both the hydride and chloride derivatives, (X=H^?, Cl^?), underwent exchange with free ¹³CO. Under reduced pressure, (R,R)‐(^iPrDuPhos)Co(CO)₂Cl underwent CO dissociation to form (R,R)‐(^iPrDuPhos)Co(CO)Cl.