The synthesis of oligomers of oxetane-based dipeptide isosteres derived from L-rhamnose or D-xylose.

Routes to oligomers (dimers, tetramers, hexamers) of five oxetane-based dipeptide isosteres have been established. Methyl 2,4-anhydro-5-azido-5-deoxy-L-rhamnonate 'monomer' led, by coupling the corresponding carboxylic acid and amine, to a 'dimer'. Reverse-aldol ring-opening occurred on attempted saponification of the dimer, so all further oligomerization was performed using TBDMS C-3 hydroxyl protection. The silyl protected L-rhamnonate monomer led in turn to the dimer (via the monomer acid and amine), the tetramer (via the dimer acid and amine) and finally the hexamer (via the tetramer acid and dimer amine). In each case the acids were obtained through saponification of the respective methyl esters and the amines were obtained by hydrogenation of the azides; coupling was TBTU-mediated. Essentially the same strategy was employed on equivalent D-lyxonate, 6-deoxy-L-altronate, 6-deoxy-D-gulonate and D-fuconate dipeptide isosteres to give the respective dimers, tetramers and hexamers.     

Oxygen consumption, ventilatory frequency and heart rate of lampreys (Lampetra fluviatilis) during their spawning run.

1. The standard rate of oxygen consumption, ventilatory frequency and heart rate of adult Lampetra fluviatilis were measured during the light phase of the photoperiod and at times corresponding to various stages in the upstream migration. 2. All three parameters increased during the spawning run but only in mature individuals were significant differences found between the sexes. 3. The regression coefficients for the logarithmic relationship between oxygen consumption and body weight of immature animals were 0.912 and 0.925 at 9.5 and 16 degrees C respectively. 4. Both the standard rate of oxygen consumption and the amount of oxygen taken up during activity increased greatly during the hours of darkness. 5. Oxygen consumption, ventilatory frequency and, to a lesser extent, heart rate increased significantly at 9.5 degrees C over the 100-20% range of saturation with air. 6. Below 20% saturation with air, lampreys no longer remained attached by their oral disc for prolonged periods and the ventilatory frequency rose even more rapidly to reach a maximum of 175 beats/min at 12.5%. Exposure to 7.5% resulted in death within 5-8 h.     

Protein-nanocrystal conjugates support a single filament polymerization model in R1 plasmid segregation

To ensure inheritance by daughter cells, many low-copy number bacterial plasmids, including the R1 drug-resistance plasmid, encode their own DNA segregation systems. The par operon of plasmid R1 directs construction of a simple spindle structure that converts free energy of polymerization of an actin-like protein, ParM, into work required to move sister plasmids to opposite poles of rod-shaped cells. The structures of individual components have been solved, but little is known about the ultrastructure of the R1 spindle. To determine the number of ParM filaments in a minimal R1 spindle, we used DNA-gold nanocrystal conjugates as mimics of the R1 plasmid. We found that each end of a single polar ParM filament binds to a single ParR/parC-gold complex, consistent with the idea that ParM filaments bind in the hollow core of the ParR/parC ring complex. Our results further suggest that multifilament spindles observed in vivo are associated with clusters of plasmids segregating as a unit.     

Fish fauna of the Severn Estuary. Are there long-term changes in abundance and species composition and are the recruitment patterns of the main marine species correlated?

Fish were collected from the intake screens of the Oldbury Power Station in the Severn Estuary in each week between early July 1972 and late June 1977 and at least twice monthly between early January 1996 and late June 1999. The annual catches, after adjustment to a common sampling effort, demonstrate that the abundance of fish at Oldbury was far greater in the 1990s than 1970s, mainly due to marked increases in the numbers of certain marine species, such as sand goby, whiting, bass, thin-lipped grey mullet, herring, sprat and Norway pout. These increases may reflect the great improvement that occurred in the water quality of the Severn Estuary between these decades. The only species that declined markedly in abundance was poor cod. Modest declines in flounder and River lamprey paralleled those occurring elsewhere in the UK. The species composition in the two decades also differed, reflecting changes not only in the relative abundances of the various marine estuarine-opportunistic species, which dominated the ichthyofauna, but also in those of the suite of less abundant species in the estuary. The cyclical changes undergone each year by the species composition of the fish fauna of the Severn Estuary reflect sequential intra-annual changes in the relative abundances of species representing each of the marine, diadromous and freshwater categories. New approaches have been developed to test whether or not large sets of correlations between patterns of recruitment amongst abundant marine species (internal correlations), and between those patterns and salinity and water temperature within the estuary (cross-correlations), were significant. The correlation profile analyses found no evidence that the annual recruitment strengths of these species were either intercorrelated, or correlated with either one or a combination of both of the above environmental variables. Yet, the timings of the recruitment of these species into the estuary were intercorrelated, i.e. a slightly earlier or later than normal immigration by one species in a given year was paralleled by the same trend in other species. However, this association in recruitment times could be linked neither to salinity nor water temperature within the estuary, nor to a combination of these two variables. These results indicate that, while the factors that influence the annual recruitment strengths of the juveniles of different marine species vary, inter-annual differences in the phasing of events that regulate spawning times and/or larval dispersal influence, in the same direction, the times when marine species are recruited into the estuary.     

Identification of a novel series of potent RON receptor tyrosine kinase inhibitors

A novel series of N-(3-fluoro-4-(2-substituted-thieno[3,2-b]pyridin-7-yloxy)phenyl)-1-phenyl-5-(trifluoromethyl)-1H-pyrazole-4-carboxamides targeting RON receptor tyrosine kinase was designed and synthesized. SAR study of the series allowed us to identify compounds possessing either inhibitory activity of RON kinase enzyme in the low nanomolar range with low residual activity against the closely related c-Met or potent dual inhibitory activity against RON and c-Met, ? with no significant activity against VEGFR2 in both cases.     

Oxidative degradation of bilirubin produces vasoactive compounds.

Subarachnoid haemorrhage is often followed by haemolysis and concomitant oxidative stress, and is frequently complicated by pathological vasoconstriction or cerebral vasospasm. It is known that upregulation of haem oxygenase (HO-1) is induced by oxidative stress and results in release of biliverdin and bilirubin (BR), which are scavengers of reactive oxygen species (ROS). Here we report biomimetic studies aimed at modelling pathological conditions leading to oxidative degradation of BR. Oxidative degradation products of BR, formed by reaction with hydrogen peroxide (an ROS model system), demonstrated biological activity by stimulating oxygen consumption and force development in vascular smooth muscle from porcine carotid artery. Analogous biological activity was observed with vasoactive cerebrospinal fluid from subarachnoid haemorrhage patients. Three degradation products of BR were isolated: two were assigned as isomeric monopyrrole (C9H11N2O2) derivatives, 4-methyl-5-oxo-3-vinyl-(1, 5-dihydropyrrol-2-ylidene)acetamide and 3-methyl-5-oxo-4-vinyl-(1, 5-dihydropyrrol-2-ylidene)acetamide and the third was 4-methyl-3-vinylmaleimide (MVM), a previously isolated photodegradation product of biliverdin. Possible mechanisms of oxidative degradation of BR are discussed. Tentative assignment of these structures in the cerebrospinal fluid (CSF) of cerebral vasospasm patients has been made. It is proposed that one or more of the degradation products of biliverdin or bilirubin are involved in complications such as vasospasm and or pathological vasoconstriction associated with haemorrhage.     

Mechanistic insights into the inhibition of serine proteases by monocyclic lactams.

Although originally discovered as inhibitors of pencillin-binding proteins, beta-lactams have more recently found utility as serine protease inhibitors. Indeed through their ability to react irreversibly with nucleophilic serine residues they have proved extraordinarily successful as enzyme inhibitors. Consequently there has been much speculation as to the reason for the general effectiveness of beta-lactams as antibacterials or inhibitors of hydrolytic enzymes. The interaction of analogous beta- and gamma-lactams with a serine protease was investigated. Three series of gamma-lactams based upon monocyclic beta-lactam inhibitors of elastase [Firestone, R. A. et al. (1990) Tetrahedron 46, 2255-2262.] but with an extra methylene group inserted between three of the bonds in the ring were synthesized. Their interaction with porcine pancreatic elastase and their efficacy as inhibitors were evaluated through the use of kinetic, NMR, mass spectrometric, and X-ray crystallographic analyses. The first series, with the methylene group inserted between C-3 and C-4 of the beta-lactam template, were readily hydrolyzed but were inactive or very weakly active as inhibitors. The second series, with the methylene group between C-4 and the nitrogen of the beta-lactam template, were inhibitory and reacted reversibly with PPE to form acyl-enzyme complexes, which were stable with respect to hydrolysis. The third series, with the methylene group inserted between C-2 and C-3, were not hydrolyzed and were not inhibitors consistent with lack of binding to PPE. Comparison of the crystal structure of the acyl-enzyme complex formed between PPE and a second series gamma-lactam and that formed between PPE and a peptide [Wilmouth, R. C., et al. (1997) Nat. Struct. Biol. 4, 456-462.] reveals why the complexes formed with this series were resistant to hydrolysis and suggests ways in which stable acyl-enzyme complexes might be obtained from monocyclic gamma-lactam-based inhibitors.     

Prolonged intermittent high intensity exercise impairs neuromuscular performance of the knee flexors

This study investigated the effect of prolonged intermittent high intensity exercise upon the isokinetic leg strength and electromechanical delay of the knee flexors. Seven male collegiate soccer players were exposed to: (i) a prolonged intermittent high intensity exercise task (PIHIET) which required subjects to complete a single-leg pedalling task, with the preferred limb, (75 rpm for all constant-load portions of the task) consisting of 48 × 1.8 minute cycles of exercise, and (ii) a control task consisting of no exercise. Pre-, mid- and post-PIHIET gravity corrected indices of knee flexion angle-specific torque (0.44 rad knee flexion (AST); 0 rad = full knee extension; [1.05 rad · s⁻¹]) were made for both intervention and control limbs. Electromechanical delay (EMD) of the m. biceps femoris during supine knee flexion movements was evaluated in the preferred leg on both intervention and control days. Repeated measures ANOVAs revealed significant condition (intervention; control) by time (pre; mid; post) interactions for both knee flexor AST (F_[2,12] = 4.8; p<0.03) and EMD (F_[2,12] = 4.1; p<0.05). AST was observed to decrease by 16% and EMD increase by 30% pre to post intervention. These observations suggest an impairment of neuromuscular control and the ability to maintain force generation in the knee flexors, near the extremes of the range of motion during prolonged intermittent high-intensity exercise activities. Changes of this magnitude may pose a threat to the integrity of the knee joint. Accepted: 6 January 1998