Light and electron microscopic distribution of the AMPA receptor subunit, GluR2, in the spinal cord of control and G86R mutant superoxide dismutase transgenic mice

The NMR structure of the 98 residue beta-elicitin, cryptogein, which induces a defence response in tobacco, was determined using 15N and 13C/15N labelled protein samples. In aqueous solution conditions in the millimolar range, the protein forms a discrete homodimer where the N-terminal helices of each monomer form an interface. The structure was calculated with 1047 intrasubunit and 40 intersubunit NOE derived distance constraints and 236 dihedral angle constraints for each subunit using the molecular dynamics program DYANA. The twenty best conformers were energy-minimized in OPAL to give a root-mean-square deviation to the mean structure of 0.82 A for the backbone atoms and 1.03 A for all heavy atoms. The monomeric structure is nearly identical to the recently derived X-ray crystal structure (backbone rmsd 0.86 A for residues 2 to 97) and shows five helices, a two stranded antiparallel beta-sheet and an omega-loop. Using 1H,15N HSQC spectroscopy the pKa of the N- and C-termini, Tyr12, Asp21, Asp30, Asp72, and Tyr85 were determined and support the proposal of several stabilizing ionic interactions including a salt bridge between Asp21 and Lys62. The hydroxyl hydrogens of Tyr33 and Ser78 are clearly observed indicating that these residues are buried and hydrogen bonded. Two other tyrosines, Tyr47 and Tyr87, show pKa's > 12, however, there is no indication that their hydroxyls are hydrogen bonded. Calculations of theoretical pKa's show general agreement with the experimentally determined values and are similar for both the crystal and solution structures.     

The use of a non‐equilibrated solid phase microextraction method to quantitatively determine the off‐notes in mint and other essential oils

The undesirable top notes or off‐notes found in mint, clary sage, and cedarwood oils could be quantitatively determined using a non‐equilibrated solid phase microextraction/gas chromatography/selected ion monitoring/mass spectrometry (SPME/GC/SIM‐MS) technique. Using the low threshold components, dimethyl sulfide, 2‐methylpropanal, 2‐methylbutanal, and 3‐methylbutanal, which are associated with the off‐notes of these oils, their levels could be reproducibly quantitatively determined. The highest level of off‐notes was found in a sample of Scotch spearmint oil where the levels of the four constituents were, dimethyl sulfide (238 µg g^?1), 2‐methylpropanal (286 µg g^?1), 2‐methylbutanal (1048 µg g^?1) and 3‐methylbutanal (1489 µg g^?1). These quantitative results in combination with sensory evaluations could provide for a powerful overall assessment of essential oil quality. Copyright © 2004 Society of Chemical Industry     

Benefits of a clinical pharmacokinetic service in optimising phenytoin use in the western Cape

OBJECTIVES: To study the benefits of a clinical pharmacokinetic service in optimising phenytoin use in the Western Cape. DESIGN: Assessment of the response to treatment was based on the number of seizures during the 3 months before entering the study (first baseline period), 3 months after entering the study (second baseline period) and 3 months before the termination of the study (test period). Patients kept a seizure diary throughout the study. The Michaelis-Menten model was used to calculate doses and predict steady-state serum concentrations. SETTING: Nine epilepsy clinics. SUBJECTS: One hundred and ninety-five (113 black and 82 coloured) compliant people with epilepsy receiving generic phenytoin monotherapy. OUTCOME MEASURES: Reduction in seizure frequency and adverse effects. RESULTS: A reduction in seizure frequency (64.8% compared with pre-optimisation) was experienced by 64.9% of patients. Mean seizure frequency was reduced from 3.39 to 1.18 per month. Reductions in seizure frequency of 100% and more than 50% were reported by 39.2% and 58.7% of patients, respectively. Adverse effects of phenytoin were reduced from 20.5% at the first visit to 3.2% at the last visit. CONCLUSION: The clinical pharmacokinetic dosing service for phenytoin applied in this study contributed significantly to the success of epilepsy management.     

Synthesis, structure, and antiproliferative activity of selenophenfurin, an inosine 5'-monophosphate dehydrogenase inhibitor analogue of selenazofurin

The synthesis and biological activity of selenophenfurin (5-beta-D-ribofuranosylselenophene-3-carboxamide, 1), the selenophene analogue of selenazofurin, are described. Glycosylation of ethyl selenophene-3-carboxylate (6) under stannic chloride-catalyzed conditions gave 2- and 5-glycosylated regioisomers, as a mixture of alpha- and beta-anomers, and the beta-2,5-diglycosylated derivative. Deprotected ethyl 5-beta-D-ribofuranosylselenophene-3-carboxylate (12 beta) was converted into selenophenfurin by ammonolysis. The structure of 12 beta was determined by 1H- and 13C-NMR, crystallographic, and computational studies. Selenophenfurin proved to be antiproliferative against a number of leukemia, lymphoma, and solid tumor cell lines at concentrations similar to those of selenazofurin but was more potent than the thiophene and thiazole analogues thiophenfurin and tiazofurin. Incubation of K562 cells with selenophenfurin resulted in inhibition of IMP dehydrogenase (IMPDH) (76%) and an increase in IMP pools (14.5-fold) with a concurrent decrease in GTP levels (58%). The results obtained confirm the hypothesis that the presence of heteroatoms such as S or Se in the heterocycle in position 2 with respect to the glycosidic bond is essential for both cytotoxicity and IMP dehydrogenase inhibitory activity in this type of C-nucleosides.     

Wear in retrieved Charnley acetabular sockets

One hundred and twenty-nine Charnley acetabular components were acquired at the time of revision surgery and a tribological investigation undertaken. The relative occurrence of pitting in the unworn and worn regions of the sockets suggest that most of the cement ingress occurs during the early part of the service life. The penetration depth of the explanted sockets was determined using the shadowgraph technique. Observation of the profiles in the wear planes suggest that, in general, the creep component was not a significant proportion of the overall change in the inner bore of the socket. Using weighted ordinary least squares regression, in which the intercept was not assumed to be zero, mean penetration and wear volume rates of 0.02 (SE = 0.02) mm/year and 55 (SE = 5) mm3/year, respectively, were recorded and are in agreement with other retrieval studies. In neither case was the intercept found to be significantly different from zero. A mean clinical wear factor, Kclinical, equal to 2.1 (SE = 0.2) x 10(-6) mm3/N m was calculated which is considerably larger than that found in laboratory experiments which purport to reflect in vivo conditions. In this analysis, a significant positive intercept was observed [96 (SE = 36) mm3] and may be evidence of the small initial penetration due to creep reported in simulator experiments. A strong positive association between kclinical and the arithmetical mean roughness, Ra, of the femoral head was also demonstrated although the rate of change was not as great as that cited for laboratory experiments.     

A role for tissue factor in cell adhesion and migration mediated by interaction with actin-binding protein 280

Tissue factor (TF), the protease receptor initiating the coagulation system, functions in vascular development, angiogenesis, and tumor cell metastasis by poorly defined molecular mechanisms. We demonstrate that immobilized ligands for TF specifically support cell adhesion, migration, spreading, and intracellular signaling, which are not inhibited by RGD peptides. Two-hybrid screening identified actin-binding protein 280 (ABP-280) as ligand for the TF cytoplasmic domain. Extracellular ligation of TF is necessary for ABP-280 binding. ABP-280 recruitment to TF adhesion contacts is associated with reorganization of actin filaments, but cytoskeletal adaptor molecules typically found in integrin-mediated focal contacts are not associated with TF. Chimeric molecules of the TF cytoplasmic domain and an unrelated extracellular domain support cell spreading and migration, demonstrating that the extracellular domain of TF is not involved in the recruitment of accessory molecules that influence adhesive functions. Replacement of TF's cytoplasmic Ser residues with Asp to mimic phosphorylation enhances the interaction with ABP-280, whereas Ala mutations abolish coprecipitation of ABP-280 with immobilized TF cytoplasmic domain, and severely reduce cell spreading. The specific interaction of the TF cytoplasmic domain with ABP-280 provides a molecular pathway by which TF supports tumor cell metastasis and vascular remodeling.     

Clinical benefits of a consistent reduction in blood pressure

This paper briefly reviews the epidemiological evidence that hypertension is a major cardiovascular risk factor. It also summarizes the data from controlled intervention trials that show antihypertensive treatment to be accompanied by a reduction in cardiovascular morbidity and mortality. The inability of antihypertensive treatment to offer full protection to the hypertensive individual is then discussed, together with the therapeutic strategies to increase the benefits, particularly with respect to limiting end-organ damage and reduction of cardiovascular events. In this context, emphasis is given to the potential additional benefit conferred by control of 24-h blood pressure and to the compliance advantage of using drugs with a long duration of action. The longevity of the blood pressure lowering effect can compensate for delayed or missed drug consumption, a frequent phenomenon in the chronically treated hypertensive patient.     

The change in employee loyalty

Employee loyalty affects company performance, competitiveness and ultimately the bottom line. Today, the old employment contract is null and void. Employee loyalty cannot be taken for granted; it must be fostered and cultivated. The new employment contract encourages participative decision making, career development, pay for productivity and flexible benefits.