A peptide derived from apomyoglobin by cyanogen bromide cleavage was found to be an active emulsifier. This molecule, peptide 1-55, has two potential amphipathic alpha-helices and a hydrophilic C-terminal domain. The importance of each of these domains to the emulsifying properties of this molecule was investigated by testing the products of gene constructs based on the sequence of peptide 1-55, but lacking one of the three domains. The emulsifying activity of the peptides lacking either of the alpha-helices was correlated with the hydrophobic moments of their respective helices. The hydrophobic moment is a measure of the amphipathicity of alpha-helices; a hydrophobic moment analysis of other emulsifying peptides supports the hypothesis that a high hydrophobic moment contributes to good emulsifying properties in a molecule which contains alpha-helices.

Amino Acid Sequence↗

PubMed2000

The classical arabinogalactan protein gene family of arabidopsis.

Arabinogalactan proteins (AGPs) are extracellular proteoglycans implicated in plant growth and development. We searched for classical AGPs in Arabidopsis by identifying expressed sequence tags based on the conserved domain structure of the predicted protein backbone. To confirm that these genes encoded bona fide AGPs, we purified native AGPs and then deglycosylated and deblocked them for N-terminal protein sequencing. In total, we identified 15 genes encoding the protein backbones of classical AGPs, including genes for AG peptides-AGPs with very short backbones (10 to 13 amino acid residues). Seven of the AGPs were verified as AGPs by protein sequencing. A gene encoding a putative cell adhesion molecule with AGP-like domains was also identified. This work provides a firm foundation for beginning functional analysis by using a genetic approach.

Amino Acid Sequence↗

PubMed2000

Are cost of injury studies useful?

Explore the source record for details and available documents.

Canada↗

PubMed1999

Limits to employee empowerment in the UK NHS. Locality managers in a community health trust.

The concern of this paper lies with empowerment of middle managers in a community health trust. It considers the impact of traditional structures and cultures upon the level of decision which middle managers are empowered to make. The conclusion reached is one which suggests that the level of discretion available to middle managers was one of making task-oriented decisions rather than decisions about strategic change. However, even this was compromised by financial constraints and by "best practices" imposed by the personnel function.

Administrative Personnel↗

PubMed1998

The glycoinositolphospholipids from Leishmania panamensis contain unusual glycan and lipid moieties.

The cell surface of Leishmania parasites is coated by glycosylphosphatidylinositol (GPI)-anchored macromolecules (glycoproteins and a lipophosphoglycan) and a polymorphic family of free GPI glycolipids or glycoinositolphospholipids (GIPLs). Here we show that GIPLs with unusual glycan and lipid moieties are likely to be major cell surface components of L. panamensis (subgenus Viannia) promastigotes. These glycolipids were purified by high performance thin layer chromatography and their structures determined by gas-liquid chromatography-mass spectrometry, fast-atom bombardment mass spectrometry, methylation analysis and chemical and enzymatic sequencing of the glycan headgroups. The major GIPLs contained two glycan core sequences, Manalpha1-3Manalpha1-4GlcN-phosphatidylinositol (type-2 series) or Manalpha1-3[Manalpha1-2Manalpha1-6]Manalpha1- 4GlcN-phosphatidylinosit ol (hybrid series), which were elaborated with Galalpha1-2Galbeta1- or Galalpha1-2/3Galalpha1-2Galbeta1- extensions that were attached to the 3-position of the alpha1-3 linked mannose. The phosphatidylinositol moiety contained exclusively diacylglycerol with palmitoyl, stearoyl and heptadecanoyl chains. Non-galactosylated GIPL species with the same core structures were also found. The galactose extensions and the presence of diacylglycerol in the lipid moieties are novel features for the GIPLs of Leishmania spp. The implications of these structures for the biosynthesis of leishmanial GIPLs and their putative function in the mammalian host are discussed.

Animals↗

PubMed1998

Stage-specific proteophosphoglycan from Leishmania mexicana amastigotes. Structural characterization of novel mono-, di-, and triphosphorylated phosphodiester-linked oligosaccharides.

Intracellular amastigotes of the protozoan parasite Leishmania mexicana secrete a macromolecular proteophosphoglycan (aPPG) into the phagolysosome of their host cell, the mammalian macrophage. The structures of aPPG glycans were analyzed by a combination of high pH anion exchange high pressure liquid chromatography, gas chromatography-mass spectrometry, enzymatic digestions, electrospray-mass spectrometry as well as 1H and 31P NMR spectroscopy. Some glycans are identical to oligosaccharides known from Leishmania mexicana promastigote lipophosphoglycan and secreted acid phosphatase. However, the majority of the aPPG glycans represent amastigote stage-specific and novel structures. These include neutral glycans ([Glcbeta1-3]1-2Galbeta1-4Man, Galbeta1-3Galbeta1-4Man, Galbeta1-3Glcbeta1-3Galbeta1-4Man), several monophosphorylated glycans containing the conserved phosphodisaccharide backbone (R-3-[PO4-6-Gal]beta1-4Man) but carrying stage-specific modifications (R = Galbeta1-, [Glcbeta1-3]1-2Glcbeta1-), and monophosphorylated aPPG tri- and tetrasaccharides that are uniquely phosphorylated on the terminal hexose (PO4-6-Glcbeta1-3Galbeta1-4Man, PO4-6-Glcbeta1-3Glcbeta1-3Galbeta1-4Man, PO4-6-Galbeta1-3Glcbeta1-3Galbeta1-4Man). In addition aPPG contains highly unusual di- and triphosphorylated glycans whose major species are PO4-6-Glcbeta1-3Glcbeta1-3[PO4-6-Gal]beta1-4Man, PO4-6-Galbeta1-3Glcbeta1-3[PO4-6-Gal]beta1-4Man, PO4-6-Galbeta1-3Glcbeta1-3Glcbeta1-3[PO4-6-Gal]beta1-+ ++4Man, PO4-6-Glcbeta1-3[PO4-6-Glc]beta1-3[PO4-6-Gal]beta1-4Man, PO4-6-Galbeta1-3[PO4-6-Glc]beta1-3Glcbeta1-3[PO4-6-Gal]beta1 -4Man, and PO4-6-Glcbeta1-3[PO4-6-Glc]beta1-3Glcbeta1-3[PO4-6-Gal]beta1 -4Man. These glycans are linked together by the conserved phosphodiester R-Manalpha1-PO4-6-Gal-R or the novel phosphodiester R-Manalpha1-PO4-6-Glc-R and are connected to Ser(P) of the protein backbone most likely via the linkage R-Manalpha1-PO4-Ser. The variety of stage-specific glycan structures in Leishmania mexicana aPPG suggests the presence of developmentally regulated amastigote glycosyltransferases which may be potential anti-parasite drug targets.

Animals↗

PubMed1998

Stakeholders' views of management development as a cultural change process in the Health Service.

Illustrates the failure of a competence-based management development programme in a hospital trust to achieve organisational objectives. The approach and content of the management development programme is resisted by participants despite the facilitators drawing on management theory to legitimate it. The question posed in this paper, building on an earlier study by Holman and Hall (1996), is whether the generic approach of competence-based management development is appropriate in the UK NHS. The theoretical inspiration for the research is what may be termed a "processual" or "contextualist" approach which is clearly influenced by "negotiated order" literature.

Health Services Research↗

PubMed1998

Managerialism in the health service: partnership or conflict in a management development programme.

This paper describes a management development programme in a hospital-based Trust and the differing responses demonstrated by those taking part. In terms of the impact of the programme, a picture emerges of differentiation and fragmentation rather than culture being managed to produce a social glue. Following this, suggestions are made for successful partnerships in the promotion of managerial effectiveness through management development to clinical services managers in the health service.

Commerce↗

PubMed1997

Identification of active-site histidine residues of a self-incompatibility ribonuclease from a wild tomato.

The style component of the self-incompatibility (S) locus of the wild tomato Lycopersicon peruvianum (L.) Mill. is an allelic series of glycoproteins with ribonuclease activity (S-RNases). Treatment of the S3-RNase from L. peruvianum with iodoacetate at pH 6.1 led to a loss of RNase activity. In the presence of a competitive inhibitor, guanosine 3'-monophosphate (3'-GMP), the rate of RNase inactivation by iodoacetate was reduced significantly. Analysis of the tryptic digestion products of the iodoacetate-modified S-RNase by reversed-phase high-performance liquid chromatography and electrospray-ionization mass spectrometry showed that histidine-32 was preferentially modified in the absence of 3'-GMP. Histidine-88 was also modified, but this occurred both in the presence and absence of 3'-GMP, suggesting that this residue is accessible when 3'-GMP is in the active site. Cysteine-150 was modified by iodoacetate in the absence of 3'-GMP and, to a lesser extent, in its presence. The results are discussed with respect to the related fungal RNase T2 family and the mechanism of S-RNase action.

Amino Acid Sequence↗

PubMed1997

Contested terrain: the incomplete closure of managerialism in the health service.

Focuses on feelings about culture and change in the health service and the impact of management development programmes on the change process. The issues raised in a series of semistructured interviews are taken forward to a case study, a medium-sized hospital Trust. The researcher "hangs around and listens into" a management development programme aimed at middle managers with a nursing background. The theme of ideological conflict comes into central focus. Analysis of outcomes suggests that the managerial assumptions on which the programme is based result in resistance from participants. They resist the attempts of facilitators to provide "closure" whereby managerial ways of doing things are suggested as the "obvious way ahead". Advocates that management education particularly in the health service should have a pluralistic orientation. As part of such pluralism, more critical approaches should be considered beyond conventional and problematic conceptions of knowledge relating to management. These are reductionist in considering a manager as "having a set of technical competences" rather than exhibiting "a way of being".

Administrative Personnel↗

PubMed1996

Structural characterisation of xyloglucan secreted by suspension-cultured cells of Nicotiana plumbaginifolia.

Linkage analysis of a xyloglucan from the extracellular medium of suspension cultures of Nicotiana plumbaginifolia showed mostly 4-Glcp and 4,6-Glcp, terminal Xylp and 2-Xylp, and terminal Araf, along with approximately 10% (w/w) O-acetyl groups, equivalent to approximately 0.28 mol acetyl per mol of glycosyl residue. Methylation with methyl trifluoromethanesulfonate under neutral conditions, followed by re-methylation with CD3I under basic conditions, and conversion into partially methylated alditol acetates showed that O-acetyl groups were primarily attached to C-6 of approximately 44% of the 4-Glcp backbone not substituted with Xylp residues and to C-5 of approximately 15% of the terminal Araf residues. These positions of the O-acetyl groups were confirmed by 1H-NMR. Oligosaccharides generated by digestion of native xyloglucan with endo-(1-->4)-beta-glucanase were separated by a combination of gel-filtration chromatography and anion-exchange HPLC, and analysed by glycosyl linkage analysis and by electrospray ionisation-mass spectrometry (ESI-MS). The major oligosaccharide subunits were Glc4Xyl2 and Glc5Xyl2, of which 50-60% are substituted with one terminal Araf residue attached to O-2 of a Xylp residue, and a further 20-25% are substituted with two terminal Araf residues attached to O-2 of the Xylp residues. ESI-MS showed that many of the oligosaccharide subunits carried one, two, and, occasionally three O-acetyl groups.

Acetylation↗

PubMed1996

Analysis of the structural heterogeneity of laminarin by electrospray-ionisation-mass spectrometry.

Electrospray-ionisation-mass spectrometry (ESIMS) was used in conjunction with chemical derivatisation and degradation procedures to analyse the size heterogeneity and branching structure of laminarin from the brown alga, Laminaria digitata. Laminarin is a beta-(1-->3)-linked D-glucan with occasional beta-(1-->6)-linked branches. Electrospray-ionisation-mass spectrometry of permethylated laminarin distinguished two homologous series of molecules, a minor G-series containing 22-28 glucosyl residues, and a more abundant M-series containing 20-30 glucosyl residues linked to a mannitol residue. The relative abundance of all these molecular species could be determined simultaneously from a single mass spectrum, with a mean mass error of 0.6 atomic mass units and a mean mass accuracy of 0.011%. Both series had a mean degree of polymerisation of 25 glucosyl residues, and an approximately 3:1 molar ratio of M-series to G-series molecules was maintained across the range of molecular sizes. Treatment of laminarin with periodate, followed by reduction with borohydride, degraded terminal glucosyl residues on both the main chain and the branches, and allowed the detection of isomers differing solely in their degree of branching. M-series molecules were thus shown to contain 0, 1, 2, 3 or 4 branches, with an average of 1.3 branches per molecule; branched G-series molecules were also detected. Subsequent treatment with acid (Smith degradation) showed that 75% of the branches were single glucosyl residues. This study thus shows how the speed, resolution and mass accuracy of electrospray-ionisation-mass spectrometry can be used in the detailed structural analysis of a polydisperse polysaccharide.

Carbohydrate Sequence↗

PubMed1996

Characterization of lipophosphoglycan from a ricin-resistant mutant of Leishmania major.

One of the virulence factors of the protozoan parasite Leishmania major is the surface glycoconjugate, lipophosphoglycan (LPG). A Ricin-resistant mutant of L.major was generated and characterised with respect to its virulence in mice and the structure and expression of LPG. The LPG from this mutant (1F6-B5) retained the tripartite structure of wild-type LPG, comprising a glycosylphosphatidylinositol (GPI) anchor linked to a phosphorylated disaccharide backbone terminating in a nonreducing neutral oligosaccharide cap. The structure of the GPI anchor and the major capping oligosaccharide were identical to wild-type LPG. However, there were variations in the number of phosphorylated repeats (PO4-6Gal(beta 1-4)Man(alpha 1-) comprising the backbone of LPG, although the degree of substitution with side branches (approx. 95%) was similar to that of wild-type LPG. Thus, the mutant LPG was shorter in length having, on average, 15 repeat units per molecule compared with 30 in the wild-type LPG. The mutant LPG contained both arabinose (Ara(beta 1-2)[Gal(beta 1-3)-]1,2) and galactose ([Gal(beta 1-3)-]1-8) capped side branches linked to the backbone. In contrast to wild-type LPG, the number of arabinose-capped side chains was significantly reduced, and a new population of galactose-capped (Gal(beta 1-3)]5-8) side branches was present. The level of LPG expression in mutant parasites was approximately one-tenth of the wild-type parasite. The mutant parasites were avirulent in mice. Over a period of 18 months, they did not cause lesions and organisms could not be isolated from the draining lymph nodes.

Animals↗