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Biomedical subjects

D H Williams

Publications and source records attributed to D H Williams.

At least 55 records · Page 3Linked to original sources
PubMed1998

Cleavage of rhamnose from ristocetin A removes its ability to induce platelet aggregation.

Ristocetin A is used clinically as a tool for the measurement of von Willebrand protein due to its ability to induce the aggregation of platelets in plasma being proportional to the concentration of von Willebrand protein. The enzymatically induced cleavage of alpha-L-rhamnose from the tetrasaccharide of ristocetin A removes this ability to induce platelet aggregation in plasma, providing a possible insight into the structural basis for this effect.

Carbohydrate Conformation↗
PubMed1998

Induction of asymmetry into homodimers.

The self-regulation of biological signalling receptors via homodimerization is discussed in relation to the symmetry changes occurring when these receptors bind their target ligand. The idea of positive and negative cooperativity between dimerization and ligand binding, mediated by changes in the symmetry of the system as a source of signalling control is considered; an analogy made with the homodimerization of a glycopeptide antibiotic, ristocetin A, which displays negative cooperativity. Finally, the regulation of the bacterial aspartate receptor and the human growth hormone receptor is discussed as a function of ligand-induced asymmetry.

Animals↗
PubMed1997

A limitation of two-state analysis for transitions between disordered and weakly ordered states.

BACKGROUND: The stability of the secondary structure of particular peptide regions is often used to investigate the involvement of the region in protein folding. When analysing the relatively small populations of associated states that are formed by weak interactions (i.e. those interactions that are comparable to thermal energies), it is common practice to characterise the associated state by a parameter that is measured when this state is highly occupied. The accuracy of this method, however, has not yet been determined. RESULTS: Using as a model the vancomycin group of antibiotics, either forming dimers or binding to cell wall precursors, we have investigated the dependence of the limiting (i.e. fully associated) chemical shifts of two protons on the equilibrium constants for the formation of the fully associated states. The chemical shift shows a large variation with the equilibrium constant for the formation of the fully associated state. CONCLUSIONS: The results demonstrate, in two systems, that a parameter representing a fully associated state (chemical shift) varies greatly with the equilibrium constant for the formation of that associated state. The results have implications for two-state analyses of populations of protein fragments in which a parameter representing the fully associated state is taken to be independent of the equilibrium constant for its formation. Using two-state analysis to determine the population of associated states of protein fragments could result in an underestimation of the population of these associated states.

Magnetic Resonance Spectroscopy↗
PubMed1997

Semiquantitation of cooperativity in binding of vancomycin-group antibiotics to vancomycin-susceptible and -resistant organisms.

The association of vancomycin group antibiotics with the growing bacterial cell wall was investigated by using the cell wall precursor analog di-N-acetyl-Lys-D-Ala-D-Ala in competition binding experiments. The affinities of the antibiotics for the -D-Ala-D-Ala-containing cell wall precursors of Bacillus subtilis ATCC 6633 (a model for vancomycin-susceptible gram-positive bacteria) and for the -D-Ala-D-Lac-containing cell wall precursors of Leuconostoc mesenteroides (a model for vancomycin-resistant strains of Enterococcus faecium and Enterococcus faecalis) were determined by a whole-cell assay. The binding of strongly dimerizing antibiotics such as eremomycin to the bacterial surface was thus shown to be enhanced by up to 2 orders of magnitude (relative to the binding in free solution) by the chelate effect, whereas weakly dimerizing antibiotics like vancomycin and antibiotics carrying lipid tails (teicoplanin) benefited less (ca. 1 order of magnitude). The affinity measured in this way correlates well with the MIC of the antibiotic, and a consequence of this is that future design of semisynthetic vancomycin-group antibiotics should attempt to incorporate chelate effect-enhancing structural features.

Anti-Bacterial Agents↗
PubMed1996

Structural elucidation of XR586, a peptaibol-like antibiotic from Acremonium persicinum.

A novel peptide, XR586, has been isolated from fermentations of Acremonium persicinum (Xenova culture collection number X21488). The structure of XR586 has been elucidated by means of NMR spectroscopy, electrospray and fast-atom bombardment MS, derivatization and enzymic digestion. It has been shown to be helical by CD measurements. XR586 shows many structural and conformational features in common with peptaibols, particularly the zervamicins. Peptaibol antibiotics are peptides, typically of 15-20 residues, containing a large proportion of alpha-aminoisobutyric acid (Aib) residues. These peptides adopt a helical conformation in solution and display anti-bacterial and toxic properties due to their ability to form pores in membranes. However, while XR586 contains several Aib residues, it lacks a terminal phenylalaninol and terminates in the sequence Phe-Gly. The lack of reduction of the penultimate residue at the C-terminus may indicate that this step is normally at the end of the biosynthetic pathway of peptaibols and occurs with cleavage of Gly. The 1H chemical shift assignments of XR586 are reported in Supplementary Publication SUP 50179 (3 pages), which has been deposited at the British Library Document Supply Centre, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1996) 313, 9 ("Deposition of data').

Acremonium↗
PubMed1996

Phosphorylation of SLP-76 by the ZAP-70 protein-tyrosine kinase is required for T-cell receptor function.

Two families of tyrosine kinases, the Src and Syk families, are required for T-cell receptor activation. While the Src kinases are responsible for phosphorylation of receptor-encoded signaling motifs and for up-regulation of ZAP-70 activity, the downstream substrates of ZAP-70 are unknown. Evidence is presented herein that the Src homology 2 (SH2) domain-containing leukocyte protein of 76 kDa (SLP-76) is a substrate of ZAP-70. Phosphorylation of SLP-76 is diminished in T cells that express a catalytically inactive ZAP-70. Moreover, SLP-76 is preferentially phosphorylated by ZAP-70 in vitro and in heterologous cellular systems. In T cells, overexpression of wild-type SLP-76 results in a hyperactive receptor, while expression of a SLP-76 molecule that is unable to be tyrosine-phosphorylated attenuates receptor function. In addition, the SH2 domain of SLP-76 is required for T-cell receptor function, although its role is independent of the ability of SLP-76 to undergo tyrosine phosphorylation. As SLP-76 interacts with both Grb2 and phospholipase C-gamma1, these data indicate that phosphorylation of SLP-76 by ZAP-70 provides an important functional link between the T-cell receptor and activation of ras and calcium pathways.

Adaptor Proteins, Signal Transducing↗
PubMed1996

Sex difference in high density lipoprotein cholesterol in six countries.

It is known that women have higher levels of high density lipoprotein (HDL) cholesterol than men. The authors examined the association between HDL cholesterol and biologic sex in 8,631 women and 10,690 men aged 45-54 years from six countries studied between 1972 and 1989. The variation in the sex difference for HDL cholesterol was significant; the smallest difference (0.06 mmol/liter) was seen in China and the largest (0.40 mmol/liter) in Canada. Adjustment for differences in body mass index, smoking, alcohol use, and heart rate reduced but did not eliminate the variability. The sex difference in HDL cholesterol levels, usually assumed to be due to biologic factors, differs across cultures and may be related to environmental factors.

Arteriosclerosis↗
PubMed1996

Blood pressure and heart rate response during exercise in men and women in the USA and Russia lipid research clinics prevalence study.

Examination of blood pressure (BP) and heart rate (HR) measurements at rest and during exercise in samples of USA and Russian middle-aged men and women show significant differences between countries for both genders. Russian men had higher resting systolic blood pressure (SBP) than USA men but lower SBP at both stages of exercise. Russian women had significantly higher resting SBP than USA women at rest and also during exercise. Russian men and women had significantly lower HR at rest and during exercise than USA men and women. Differences between countries were also noted for mean body weight, height, lipid levels, percentages of smokers and several other variables, and it was postulated that differences in these factors might be responsible in part for differences in SBP and HR during exercise. After adjustment for these variables, differences in SBP during exercise between USA and Russian men remained, but differences between USA and Russian women disappeared. For both genders, differences in HR during exercise remained after adjustment. Mortality analyses in USA and Russian men indicated that stage 2 SBP response during exercise was generally not a significant mortality risk factor after adjustment for age, BMI, TC, smoking and resting SBP. In a similar model, stage 2 HR response during exercise was also a non-significant risk factor in mortality.

Adult↗
PubMed1996

Anthropometric determination of thigh volumes and thigh forces following acute training of increasing intensity in adult men.

The purpose of the present study was to investigate the influence of submaximal training of increasing intensity on the rate of fatigue and on the anthropometry of the quadriceps muscle. A group of 24 middle-aged male subjects trained three times a week for 12 weeks at incremental exercise intensities which elicited 66%-83% of the maximal heart rate reserve; 11 male subjects acted as controls. A purpose-built, isometric chair with an adjustable force transducer ensured a standard posture for each subject during a 20 s maximal voluntary contraction (MVC). The muscle plus bone mass of the thigh was calculated from a previously validated anthropometric method reinforced by measurements derived from water displacement and ultrasonics. Training-induced submaximal relationships were assessed by means of two ergometer tests of progressive intensity. Following training, the subjects' blood lactate concentrations had decreased from 5.5 (SD 1.7) to 3.9 (SD 1.4) mmol.1(-1) (P < 0.01) at the final exercise intensities. They increased thigh volumes from 10.7 (SD 1.7) to 11.0 (SD 1.7) 1 (P < 0.05) and thigh forces in 47.5% (P < 0.05) of the measurements. Heart rates and rates of perceived exertion were reduced (P < 0.01) following training; for the controls, all the above parameters remained constant. These results would seem to indicate that a submaximal type of training could enable a group of middle-aged men to increase the isometric forces of the thigh muscles by almost 50% when completing a MVC test following training. This suggests that this is a more suitable, safer form of maintaining and increasing maximal thigh forces for this, and older, age groups than specific isometric training alone.

Adult↗
PubMed1996

Substrate specificity and inhibitor profile of human recombinant p56lck from a baculovirus expression vector.

p56lck, a member of the src family of non-receptor protein receptor kinases, is required for normal signal transduction through the T cell receptor. Inappropriate T cell activation and proliferation has been identified as an early event in auto-immune disease-agents which control T cell activation through modulation of p56lck kinase activity could therefore be potential therapeutic agents for a range of pathological conditions. To identify p56lck inhibitors, we have established an assay system suitable for the high throughput screening of compound libraries. The assay uses enzyme purified from baculovirus infected SF9 cells, and a novel peptidic substrate identified by L. Cantley from a degenerate combinatorial peptide library. We have used this assay system to screen a number of different compounds as potential inhibitors of p56lck. In addition, peptides based on the substrate sequence were also tested to identify a sequence that could be used in the rational design of peptide inhibitors of p56lck.

Amino Acid Sequence↗
PubMed1996

Factors affecting superovulation in heifers treated with PMSG.

In this study we determined 1) if the immunoneutralization of PMSG affected the ovulatory response, the number of large follicles and embryo yield compared with that of PMSG alone or pFSH, and 2) whether the stage of the estrous cycle at which PMSG was injected affected the ovulatory response and yield of embryos in superovulated heifers. Estrus was synchronized in 99 (Experiment 1) and 71 (Experiment 2) heifers using prostaglandin F2alpha (PG) analogue, cloprostenol, given 11 d apart in replicate experiments over 2 yr. In Experiments 1 and 2, heifers were randomly allocated to 1 of 3 treatments (initiated at mid-cycle): Treatment 1--24 mg of pFSH (Folltropin) given twice daily for 4 d; Treatment 2--a single injection of 2000 IU PMSG; Treatment 3--2000 IU PMSG followed by 2000 IU of Neutra-PMSG at the time of first insemination. In Experiment 3, 116 heifers were given 2000 IU PMSG on Day 2 (n = 28), Day 3 (n = 27), Day 10 (n = 41) or Day 16 (n = 20) of the estrous cycle. The PG was given at 48 h (500 microg cloprostenol) and 60 h (250 microg cloprostenol) after the first gonadotropin treatment. Heifers were inseminated twice during estrus, and embryos were recovered on Day 7, following slaughter and graded for quality. The numbers of ovulations and large follicles (> or =10 mm) were also counted. There was no effect of treatment on ovulation rate in Experiment 1, but in Experiment 2 it was greater (P < 0.002) in heifers given PMSG (14.7 +/- 1.5) than pFSH (7.5 +/- 1.4) or PMSG-neutra-PMSG (8.7 +/- 1.5). The number of large follicles was higher following PMSG than pFSH treatment in Experiment 1, and it was higher (P < 0.004) in heifers given PMSG (5.5 +/- 0.8) than pFSH (1.12 +/- 0.7) or PMSG-neutra-PMSG (2.7 +/- 0.8) in Experiment 2. The use of Neutra-PMSG did not affect the numbers of embryos recovered or numbers of Grade 1 or 2 embryos, but it did decrease the number of Grade 3 embryos in both experiments. In Experiment 3, the ovulation rate decreased (P < 0.004) when PMSG was given on Day 3 (5.7 +/- 1.46) of the cycle rather than on Day 2 (12.3 +/- 1.64), Day 10 (13.4 +/- 1.45) or Day 16 (12.5 +/- 1.87). There was no effect of day of treatment on the numbers of large follicles. The mean numbers of embryos recovered were lower (P < 0.01) in heifers treated on Day 3 (2.1 +/- 0.67) than on Day 2 (6.8 +/- 1.0), Day 10 (6.4 +/- 0.86) or Day 16 (7.8 +/- 1.87). It is concluded that Neutra-PMSG given to heifers treated with PMSG did not improve embryo yield or quality and that treatment with PMSG early in the cycle can result in acceptable embryo yields provided sufficient time elapses between treatment and luteolysis.

Journal Article↗
PubMed1996

Weak interactions and lessons from crystallization.

Ideas derived from the study of the process of crystallization may provide insights into molecular recognition in biological systems. Both processes exploit the cooperativity which arises from the formation of a large array of weak interactions.

Anti-Bacterial Agents↗
PubMed1996

Cooperativity and anti-cooperativity between ligand binding and the dimerization of ristocetin A: asymmetry of a homodimer complex and implications for signal transduction.

BACKGROUND: Recent work has indicated that dimerization is important in the mode of action of the vancomycin group of glycopeptide antibiotics. NMR studies have shown that one member of this group, ristocetin A, forms an asymmetric dimer with two physically different binding sites for cell wall peptides. Ligand binding by ristocetin A and dimerization are slightly anti-cooperative. In contrast, for the other glycopeptide antibiotics of the vancomycin group that have been examined so far, binding of cell wall peptides and dimerization are cooperative. RESULTS: Here we show that the two halves of the asymmetric homodimer formed by ristocetin A have different affinities for ligand binding. One of these sites is preferentially filled before the other, and binding to this site is cooperative with dimerization. Ligand binding to the other, less favored half of the dimer, is anti-cooperative with dimerization. CONCLUSIONS: In dimer complexes, anti-cooperativity of dimerization upon ligand binding can be a result of asymmetry, in which two binding sites have different affinities for ligands. Such a system, in which one binding site is filled preferentially, may be a mechanism by which the cooperativity between ligand binding and dimerization is fine tuned and may thus have relevance to the control of signal transduction in biological systems.

Binding Sites↗
PubMed1996

Satisfaction with prescriptive authority protocols.

OBJECTIVES: The use of prescriptive authority protocols by pharmacists and physicians in Washington was described as to: (1) types of decisions made, frequency of use, and nature of working arrangements among participants; (2) their perceived impact on professional practice and care rendered to patients; and (3) their satisfaction with the protocol arrangements. DESIGN: A mail survey of participants currently engaged in prescriptive authority was conducted. RESULTS: Responses were received from participants in 44 of 57 protocols. A total of 135 prescribers and 84 pharmacists responded. On average, 7 pharmacists and 27 prescribers were involved in each protocol, and had been involved for 6 years or more. It was estimated that approximately 10% of all active practitioners participated in protocols. Most protocols involved continuation of drug therapy or authorizing renewals. Modification of therapy (usually involving dosage or dose from changes) was practiced in two-thirds of the protocols, and initiation of therapy in about half of them. Most existed within managed care and group practice settings. Most prescribers (98%) and pharmacists (95%) were satisfied with protocol arrangements. Additionally, they generally agreed that the protocols increase patient convenience and increase the quality of patient care, and they encouraged their colleagues to use prescribing protocols. CONCLUSIONS: Protocol arrangements appear to be working well from the perspective of participants. Based on this experience, the authors encourage the development of similar protocol arrangements in other states.

Clinical Protocols↗
PubMed1996

Increases in vocalization and motor reflex thresholds generated by the intrathecal administration of serotonin or norepinephrine.

The capacity of serotonin and norepinephrine to elevate the thresholds of spinal motor reflexes (SMRs), vocalizations during shock (VDSs), and vocalization afterdischarges (VADs) when administered into the spinal subarachnoid space was evaluated. Both monoamines generated dose-dependent increases in the thresholds of all 3 responses. The minimum effective doses of serotonin and norepinephrine that elevated all 3 response thresholds were 40 micrograms and 1 microgram respectively. Monoamine-induced increases in response thresholds were reversed by the intrathecal administration of their corresponding receptor antagonists (phentolamine or methysergide). Threshold increases generated by serotonin were also partially reduced by phentolamine. These results indicate that dorsal horn neurons that underlie flexion reflex generation (SMR) and the rostral transmission of pain information (VDS and VAD) have similar thresholds of inhibition to spinopetal monoaminergic projections.

Afferent Pathways↗
PubMed1996

Native-like beta-hairpin structure in an isolated fragment from ferredoxin: NMR and CD studies of solvent effects on the N-terminal 20 residues.

The conformational properties of protein fragments have been widely studied as models of the earliest initiation events in protein folding. While native-like alpha-helices and beta-turns have been identified, less is known about the factors that underly beta-sheet formation, in particular beta-hairpins, where considerably greater long-range order is required. The N-terminal 20 residue sequence of native ferredoxin I (from the blue-green alga Aphanothece sacrum) forms a beta-hairpin in the native structure and has been studied in isolation by NMR and CD spectroscopy. Local native-like interactions alone are unable to stabilize significantly a folded conformation of the 20-residue fragment in purely aqueous solution. However, we show that the addition of low levels of organic co-solvents promotes formation of native-like beta-hairpin structure. The results suggest an intrinsic propensity of the peptide to form a native-like beta-hairpin structure, and that the organic co-solvent acts in lieu of the stabilizing influence of tertiary interactions (probably hydrophobic contacts) which occur in the folding of the complete ferredoxin sequence. The structure of the isolated hairpin, including the native-like register of interstrand hydrogen bonding interactions, appears to be determined entirely by the amino acid sequence. The solvent conditions employed have enabled this intrinsic property to be established.

Amino Acid Sequence↗
PubMed1995

Isolation, purification and structure of exochelin MS, the extracellular siderophore from Mycobacterium smegmatis.

The extracellular siderophore from Mycobacterium smegmatis, exochelin MS, was isolated from iron-deficiently grown cultures and purified to > 98% by a combination of ion-exchange chromatography and h.p.l.c. The material is unextractable into organic solvents, is basic (pI = 9.3-9.5), has a lambda max at 420 nm and a probable Ks for Fe3+ of between 10(25) and 10(30). Its structure has been determined by examination of desferri- and ferri-exochelin and its gallium complex. The methods used were electrospray-m.s. and one- and two-dimensional (NOESY, DQF-COSY and TOCSY) 1H n.m.r. The constituent amino acids were examined by chiral g.l.c analysis of N-trifluoroacetyl isopropyl and N-pentafluoropropionyl methyl esters after hydrolysis, and reductive HI hydrolysis, of the siderophore. The exochelin is a formylated pentapeptide: N-(delta-N-formyl,delta N-hydroxy-R-ornithyl) -beta-alaninyl-delta N-hydroxy-R-ornithinyl-R-allo-threoninyl-delta N-hydroxy-S-ornithine. The linkages involving the three ornithine residues are via their delta N(OH) and alpha-CO groups leaving three free alpha-NH2 groups. Although there are two peptide bonds, these involve the three R (D)-amino acids. Thus the molecule has no conventional peptide bond, and this suggests that it will be resistant to peptidase hydrolysis. The co-ordination centre with Fe3+ is hexadenate in an octahedral structure involving the three hydroxamic acid groups. Molecular modelling shows it to have similar features to other ferric trihydroxamate siderophores whose three-dimensional structures have been established. The molecule is shown to have little flexibility around the iron chelation centre, although the terminal (Orn-3) residue, which is not involved in iron binding except at its delta N atom, has more motional freedom.

Amino Acid Sequence↗