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Biomedical subjects

G E Chapman

Publications and source records attributed to G E Chapman.

At least 19 recordsLinked to original sources

Clearance of Rh D-positive red cells with monoclonal anti-D.

Two human monoclonal antibodies, one IgG3 and one IgG1, with anti-Rh D specificity, were tested for their ability to clear red cells. Samples of red cells from 12 D-positive subjects were sensitised in vitro with various amounts of antibody, the number of antibody molecules bound to the cells was estimated, and the cells were reinjected into the donor's circulation. Both antibodies mediated clearance but substantially fewer IgG3 than IgG1 antibody molecules were required to produce a given rate of clearance. The IgG3 antibody was slightly more effective than polyclonal anti-D, as judged by comparison with previously published results. Since there is believed to be an association between the rate of red cell clearance by anti-D and the ability of the antibody to suppress D immunisation, the IgG3 monoclonal antibody may prove suitable for the immunoprophylaxis of Rh D haemolytic disease.

Antibodies, Monoclonal

Effect of biosynthetic human growth hormone on insulin action in individual tissues of the rat in vivo.

Excessive endogenous production or exogenous administration of human growth hormone (hGH) causes insulin resistance at both the hepatic and extrahepatic levels. However, which extrahepatic tissues are involved have not been defined. We have examined the diabetogenic action of authentic biosynthetic hGH on whole body glucose disposal, hepatic glucose output, and glucose metabolism in individual peripheral tissues. The use of a highly purified preparation of the hormone allowed us to examine the isolated effects of 22K hGH. The euglycemic hyperinsulinemic (approximately 100 mU/L) clamp plus 3H-2-deoxyglucose technique was used to quantitate the effects of hGH on insulin action in vivo. Administration of biosynthetic hGH at a dose of 10 IU/kg/24 h for 48 hours in male Wistar rats (approximately 340 g) produced a highly significant decrease in the steady state clamp glucose infusion rate (GIR) when compared with controls (8.1 +/- 0.6 v 18.7 +/- 0.7 mg/kg/min, P less than .001), reduced insulin-mediated suppression of hepatic glucose output (Ra) (3.9 +/- 0.6 v 0.7 +/- 0.3 mg/kg/min, P less than .05) and a decreased clamp glucose disposal rate (Rd) (12.0 +/- 0.4 v 18.10 +/- 1.1 mg/kg/min, P less than .001). There was a significant decrease in insulin-mediated glucose uptake as indicated by tissue accumulation of [3H]-2-deoxyglucose phosphorylation in diaphragm and hindlimb muscles. Insulin action was more substantially reduced in muscles (approximately 50%) than in adipose tissues (approximately 20%). These studies confirm that the diabetogenic action of hGH in the rat is due to a combination of inhibition of insulin suppression of hepatic glucose output and inhibition of the uptake and subsequent utilization of glucose in skeletal muscles.

Adipose Tissue

Partial amino acid sequence of osteocalcin from an extinct species of ratite bird.

Osteocalcin the major gamma carboxyglutamic acid containing protein of vertebrate bone has been purified from the bones of a specimen of Pachyornis elephantopus, a species of the extinct class of New Zealand ratite birds, the moas. The sequence of the N-terminal region of moa osteocalcin was determined using gas phase N-terminal sequencing. The N-terminal sequences of the ostrich and rhea osteocalcins were also determined. Alignment of the N-terminal sequence of osteocalcin from the extinct moa against the osteocalcins of the extant ostrich, rhea and emu reveals the homology amongst the ratite species is greater than the homology with the chicken osteocalcin.

Amino Acid Sequence

The response to treatment with pituitary growth hormone: the New Zealand experience.

Ninety-four growth hormone deficient New Zealand children were treated with thrice weekly intramuscular injections of human pituitary growth hormone (GH) from 1979 to 1985 and the results from 43 children were analysed in detail. Their height velocity was mean (SD) 4.0 (1.9) cm/yr immediately prior to treatment and 8.5 (3.1) cm/yr in the first year of treatment. In each of the first three years of treatment the height velocity was significantly increased (p less than 0.001), but there was a significant decline in height velocity between the first and second, and second and third years. Nevertheless each year of treatment gave a highly significant gain in relative height. Bone age did not advance more rapidly than chronological age during the first two years of treatment, but did so subsequently. The response to GH, as assessed by the increase in relative height, was negatively correlated with the relative height at the start of treatment and with the maximum pretreatment plasma GH response to insulin hypoglycaemia. There was no correlation with pretreatment relative height velocity or aetiology of GH deficiency. Height velocity improved in five of 14 children treated with thyroxine after starting GH. Discontinuation of GH for one year in 34 children resulted in a highly significant reduction in growth rate from 7.5 (2.4) cm/yr to 2.5 (2.0) cm/yr, however four prepubertal children continued to grow at a rate exceeding 4 cm/yr.

Age Determination by Skeleton

Determination of the effect of acetylation of specific lysine residues in human growth hormone on its affinity for somatogenic receptors by an affinity selection technique.

A technique is described to study the effect of acetylation of individual lysine residues in peptide hormones on the affinity for their receptors, and is illustrated for the case of human growth hormone (hGH) binding to somatogenic receptors. The hGH was partially acetylated with high specific activity [3H]-acetic anhydride and the product ([3H]-Ac-hGH) was incubated with solubilised affinity-purified somatogenic receptors (from male rat liver) in the presence and absence of excess unlabelled hGH. The receptor-bound and unbound labelled hormone were separated by gel filtration and subjected to HPLC tryptic peptide mapping after the addition of cold carrier Ac-hGH. Peaks of [3H] radioactivity were assigned to peptides corresponding to the acetylation of specific lysine residues in the hGH sequence by amino acid analysis and sequencing. Comparison of the relative intensities of corresponding [3H] peaks in the peptide maps of added receptor, bound and unbound [3H]-Ac-hGH, enabled the relative receptor-binding potencies of different acetylated hGH species to be determined. Acetylation of lysine 168 or 172 in hGH greatly decreases its receptor-binding affinity, acetylation of lysine 115 probably causes a minor decrease, whereas acetylation of lysines 38, 70, and the N-terminal amino group have no appreciable effect. Acetylation of lysine 140 causes a significant increase in receptor-binding affinity.

Acetylation

Stimulation of brain Na,K-ATPase by norepinephrine but not taurine.

The effect of taurine on rat and hamster brain Na,K-ATPase was examined and compared to norepinephrine (NE) stimulation of the enzyme. Although NE stimulation of microsomal Na,K-ATPase was observed in the presence of the cell cytosolic fraction, taurine was without effect in the presence and absence of this fraction. Taurine also failed to modulate pubescent and mature hamster brain Na,K-ATPase. Presence or absence of ion chelators did not change taurine's effect. These results are discussed in relation to previous reports of taurine and catecholamine stimulation of Na,K-ATPase.

Animals

Human growth hormone binds to lactogenic receptors in bovine, ovine and rat adrenals.

The distribution of 125I radioactivity in the liver, kidneys, adrenals and serum of male rats was measured 10 minutes after an intravenous bolus of 125I-labelled human growth hormone (hGH) was administered in the presence or absence of a large excess of ovine growth hormone or ovine prolactin. The hGH binding sites in the adrenals had displacement properties characteristic of lactogenic receptors, whereas those in the liver had displacement properties characteristic of somatogenic receptors. Bovine and ovine adrenal microsomal membrane fractions contained high affinity (Ka = 1.4-3.3 nM-1) binding sites for hGH which showed ligand specificity typical of lactogenic receptors. It is concluded that the hGH binding site in the adrenal gland is a classical lactogenic receptor and that this tissue is a convenient and rich (42.6 +/- 6.4 fmol hGH specifically bound/mg protein) source of receptor suitable for further characterization.

Adrenal Glands

Reporting therapeutic discourse in a therapeutic community.

Research in nurses' communications has concentrated on nurse to patient interactions. Those few studies which focus on nurse to nurse communications seem to be generated by a pragmatic and normative concern with effective information sharing. In this paper, which describes one aspect of a larger case study of a hospital-based therapeutic community, the description and analysis of nurses' reports flows not from a normative model of professional practice, but rather an exploration of how professional practice is articulated as discourse in nurses' written accounts. Foucault's ideas about therapeutic discourse inform the theoretical framework of the research. Ethnomethodological concerns with the importance of documentary analysis provide the methodological rationale for examining nurses' 24-hour report documents, as official discourse, reflecting therapeutic practice in this setting. A content analysis of nurses' reports, collected over a period of 4 months, demonstrated the importance of domesticity and ordinary everyday activities in nurses' accounts of hospital life. Disruption to the 'life as usual' domesticity in the community seemed to be associated with admission to and discharge from the hospital when interpersonal and interactional changes between patients occur. It is suggested that nurses in general hospital wards and more orthodox psychiatric settings might usefully consider the impact of admissions and discharges on the group of patients they manage, and make this a discursive focus of their work.

Communication

Methionine oxidation in human growth hormone and human chorionic somatomammotropin. Effects on receptor binding and biological activities.

The oxidation of the methionine residues of human growth hormone (hGH) and human chorionic somatomammotropin (hCS) to methionine sulfoxide by hydrogen peroxide has been studied. The kinetics of oxidation of individual methionine residues has been measured by reverse-phase high pressure liquid chromatography tryptic peptide mapping. Met-170 is completely resistant to oxidation in both hormones. The other 3 methionine residues in hCS (Met-64, Met-96, and Met-179) have markedly different reaction rates. Oxidation of the methionine residues does not appear to cause gross conformational changes in either hGH or hCS, as judged by CD and 1H NMR spectroscopy. Oxidation of Met-14 and Met-125 in hGH has little effect on affinity of the hormone for lactogenic receptors or on its potency in the Nb2 rat lymphoma in vitro bioassay for lactogenic hormones. The oxidation of Met-64 and/or Met-179 in hCS reduces profoundly both its affinity for lactogenic receptors and its in vitro biological potency. It is inferred by induction that residues 64 and/or 179 are critical for the binding of both hGH and hCS to lactogenic receptors and the expression of lactogenic biological activity.

Animals

The amino acid sequence of Emu osteocalcin: gas phase sequencing of Gla-containing proteins.

Osteocalcin (OC), the major gamma carboxyglutamic acid (Gla)-containing protein of vertebrate bone, has been isolated from bones of the emu (Dromaius novaehollandae) and the primary structure determined by a combination of gas phase N-terminal sequencing of the intact molecule and a proteolytic fragment, and carboxypeptidase Y C-terminal sequencing. Gla residues were located by counting tritium radioactivity in fractions from the N-terminal sequencing of the tritiated/thermally decarboxylated molecule. Emu OC consists of 48 amino acid residues containing 3 Gla residues, and a single disulphide bond. The C-terminal 29 residues are identical to those of the human and sheep OC sequences. Alignment of the N-terminal sequence against those of other OCs reveals greater sequence homology with chicken OC than with mammalian OCs.

1-Carboxyglutamic Acid

Low serum osteocalcin levels in glucocorticoid-treated asthmatics.

Serum osteocalcin (OC) levels were measured in 19 asthmatic patients receiving long term glucocorticoid therapy and in age- and sex-matched asthmatic patients not receiving this treatment. In the glucocorticoid-treated patients, the mean OC level was approximately 50% less than that in the control group (P less than 0.001), and there was a direct correlation between serum OC and 1,25-dihydroxyvitamin D [1,25-(OH)2D; r = 0.71; P less than 0.001]. Multiple regression analysis in a total of 39 glucocorticoid-treated patients indicated that OC correlated directly to 1,25-(OH)2D and inversely to glucocorticoid dose. There was no correlation between OC and 1,25-(OH)2D in the control group and no significant difference in mean serum 1,25-(OH)2D between the steroid-treated asthmatic patients and the asthmatic control patients. The effect of a 4-day course of oral 1,25-(OH)2D on serum OC was studied in six patients with glucocorticoid excess and six normal subjects. There was a similar percent increase in OC levels in both groups, though the basal concentrations and absolute increases were substantially less in the steroid-treated group. It is likely that the depression of serum OC in glucocorticoid-treated patients results from the reduction in the rate of bone formation induced by these hormones.

Administration, Oral

Immunochemical detection and characterisation of osteocalcin from moa bone.

Osteocalcin (the 6,000 dalton Mr gamma-carboxyglutamate-containing protein of bone) has been detected in acid extracts of bones of the extinct class of New Zealand ratite birds, the moas, using a radioimmunoassay for sheep osteocalcin. The immunoreactive osteocalcin of the extracts of two of these bones (the fibulae from two specimens of Pachyornis elephantopus found in South Island swamps) has been fractionated by gel filtration chromatography and reversed-phase high performance liquid chromatography, and behaves in a manner characteristic of osteocalcin from modern bones. Carbon-14 dating of bones and gizzard contents found in association with these specimens indicates approximate ages of 3,600 and 7,400 years respectively.

Animals

Population studies: an integrated course in epidemiology and sociology for medical students.

At the Royal Free Hospital School of Medicine, London, an integrated course in epidemiology and sociology for preclinical students was introduced in 1979. The course--Population Studies--is taken by the 100 second-year medical students in the summer term before they enter their clinical years. It occupies one full day and one half day each week for 8 weeks--approximately 80 hours of tuition. Population Studies is unusual in two respects. Firstly, it introduces a substantial amount of epidemiology into the preclinical curriculum. And, secondly, this is the only London medical school to integrate the teaching of sociology and epidemiology into the one course.

Education, Medical, Undergraduate

The amino acid sequences of goat, pig and wallaby osteocalcins.

The amino acid sequences of osteocalcins from cortical long bones of goat, pig and wallaby have been determined by manual microsequencing methods. The location of the gamma-carboxyglutamic acid residues were identified by N-terminal degradation of peptides in which these residues had been tritium-labelled in the gamma-H position and subsequently thermally decarboxylated to glutamic acid residues. Glutamic-17 is not gamma-carboxylated in wallaby osteocalcin, unlike other osteocalcins so far sequenced (except human). This may be related to sequence changes in the flanking residues. Proline-9 is fully hydroxylated in wallaby osteocalcin, despite the sequence substitution Ala-8----Phe-8.

Amino Acid Sequence

A proton-nuclear-magnetic-resonance study of human somatotropin (growth hormone). Assignment and properties of the histidine residues.

The 1H-n.m.r. spectra of human somatotropin (growth hormone) show perturbed peaks from individual aromatic and aliphatic apolar residues, characteristic of a specifically folded globular structure. The imidazole C-2-H resonances of the histidine residues (at positions 18, 21 and 151 in the somatotropin sequence) were individually resolved, and their titration behaviour in the pH range 1.2-11.5 was investigated. The imidazole C-2-H resonance of histidine-151 is assigned, by comparison of its titration behaviour in human somatotropin and desamido-somatotropin (Asn-152 leads to Asp-152). The C-2-H resonances of all three histidine residues are assigned, by comparison of their relative deuterium-exchange rates (determined by n.m.r.) and the relative tritium-exchange rates of the histidine residues (determined by tryptic digestion of tritiated human somatotropin and reversed-phase high-pressure liquid-chromatographic separation of the histidine-containing tryptic peptides). There is evidence that histidine-18 forms an ion-pair bond with a glutamic acid or aspartic acid residue. The globular structure does not appear to change from pH3 to 11.5, though there is evidence for an unfolding of a region of the structure (involving histidine-21 and a tyrosine residue) below pH3.

Chemical Phenomena

Ritual and rational action in hospitals.

Menzies argues that nursing hierarchies and ritual practices protect nurses from the anxieties provoked by encountering human suffering. This proposition is examined with particular reference to ritual practices in nursing. It is argued that Menzies studied nurses in isolation from the societal and subcultural norms and values which direct hospital activity. Her psychodynamic model is contrasted with a sociological model of human conduct and action. The characteristics of ritual and rational action, and the difference between non-rational and irrational rituals, is explored. The findings of three 5-month periods of participant observation are presented as illustrative case material to support the authors view, that ritual procedures are not only defence mechanisms against anxiety, but social acts which generate and convey meaning. Ritual practices described in this analysis include rituals surrounding birth, death, status and power. It is concluded that if nurses wish to change or alter ritual nursing practices in hospital it is necessary to understand their social as well as their psychological meaning.

Anxiety

The 20,000 molecular weight variant of human growth hormone. Preparation and some physical and chemical properties.

Extracts of frozen human pituitary glands contain, in addition to normal growth hormone (hGH (Mr = 22,005), a variant of lower molecular weight (approximately 20,000) (Singh, R. N. P., Seavey, B. K. and Lewis, U. J. (1974) Endocrinol. Res. Commun. 1, 449-464). This material, designated 20K hGH, occurs in various forms with the majority present as a heterologous dimer of Mr = 20,000 hGH and hGH. The purification scheme for obtaining the variant from these fractions is described. Analysis of Mr = 20,000 hGH in the sequenator has confirmed that residues 32-46 of hGH are absent in Mr = 20,000 hGH. The structures of hGH and Mr = 20,000 hGH have been compared by 1H nuclear magnetic resonance spectroscopy, circular dichroism, spectrophotometric titration, and nitration of the tyrosine residues by tetranitromethane. The results indicate that the folding of the polypeptide chain of the two proteins is similar but not identical.

Amino Acid Sequence