PubMed Health⌕ Search

Biomedical subjects

C Goddard

Publications and source records attributed to C Goddard.

At least 73 records · Page 4Linked to original sources
PubMed1988

Identification of N-myristoylated proteins by reverse-phase high performance liquid chromatography of an azlactone derivative of N-myristoylglycine.

A method is presented for the identification of N-myristoylated proteins. N-Myristoyl transferases have an absolute requirement for a free N-terminal glycine. N-Myristoylglycine is released upon mild acid hydrolysis of myristoylated peptides and proteins and its derivitization to a p-nitrobenzylazlactone with subsequent analysis by reverse phase h.p.l.c. enabled its detection to pmol levels. This facilitated the identification of N-terminal myristate in nmol quantities of purified proteins. Using this method we demonstrate that the alpha-subunit of the GTP-binding protein G0 is N-terminally myristoylated.

Chromatography, High Pressure Liquid↗
PubMed1988

Chemical and immunological characterization of the 21-kDa ADP-ribosylation factor of adenylate cyclase.

The ADP-ribosylation factor (ARF) is a 21-kDa GTP-binding protein cofactor in the cholera toxin-catalyzed ADP-ribosylation of the stimulatory regulatory subunit of adenylate cyclase. Purified bovine brain ARF was digested with cyanogen bromide, and peptides were purified and sequenced. Approximately 25-30% of the protein was sequenced in this manner. Peptides contained consensus sequences for GTP-binding proteins but were distinct from any of the previously published GTP-binding proteins. Antibodies were raised in rabbits against both protein and synthetic peptide fragments of ARF. Specific ARF immunoreactivity was detected in every eukaryotic tissue or cell examined, including yeast, slime mold, and man. No ARF immunoreactivity was observed when Escherichia coli proteins were tested. Immunoblotting revealed the majority of ARF to be present in the 100,000 x g supernatant. Immunological cross-reactivity with the cytosolic factor indicate that it and ARF are likely to be the same protein. ARF is shown to be myristylated at the amino terminus. The potential role of myristylation in cellular localization is discussed.

ADP-Ribosylation Factors↗
PubMed1988

N-myristoylation of p60src. Identification of a myristoyl-CoA:glycylpeptide N-myristoyltransferase in rat tissues.

A 16-residue synthetic peptide corresponding to the N-terminal sequence of p60src was used as the acyl acceptor in an assay for myristoyl-CoA:glycylpeptide N-myristoyltransferase in rat tissues. An additional C-terminal tyrosine amide was added to this peptide to facilitate radioiodination and enhance detectability. Reverse-phase h.p.l.c. enabled the simultaneous detection and quantification of the peptide substrate and its N-myristoylated product. N-Myristoyltransferase activity was highest in the brain with decreasing activities in lung, small intestine, kidney, heart, skeletal muscle and liver. Brain activity was distributed approximately equally between the 100,000 g pellet and supernatant fractions. The soluble enzyme exhibited a Kappm of 20 microM for the src peptide and an optimum between pH 7.0 and 7.5. Maximum N-acylating activity was seen with myristoyl (C14:0)-CoA with lower activities found with the C10:0-CoA and C12:0-CoA homologues. No activity was obtained with palmitoyl (C18:0)-CoA but this derivative inhibited N-myristoyltransferase activity greater than 50% at equimolar concentrations with myristoyl-CoA. With a decapeptide corresponding to the N-terminal sequence of the cyclic AMP-dependent protein kinase catalytic subunit as the acyl acceptor, the brain enzyme displayed a Kapp.m of 117 microM and was about 14-fold less catalytically effective than with the p60src acyl acceptor. Transferase activity was also seen with a 16-residue peptide corresponding to the N-terminal sequence of the HIV p17gag structural protein. Inhibition studies with shorter src peptide analogues indicated an enzyme specificity for the p60src acyl acceptor beyond 9 residues.

Acyltransferases↗
PubMed1988

The relationship between insulin-like growth factor-1, growth hormone, thyroid hormones and insulin in chickens selected for growth.

The concentrations of circulating insulin-like growth factor I, growth hormone, insulin and thyroid hormones were measured in broilers selected for an increase in growth, broilers in which selection pressure was relaxed and in White Leghorns. Growth hormone levels increased in all lines between 3 and 4 weeks of age followed by a decline to adult levels. The lines with the slowest rate of growth had the highest growth hormone concentrations. Insulin-like growth factor I concentrations increased significantly in all three lines of birds during the 10 weeks of study and was significantly correlated with the increase in body weight. There were no consistent differences in plasma IGF-1 levels between the lines. Thyroxine levels increased consistently throughout the study but the levels of triiodothyronine decreased between 5 and 6 weeks of age in all lines. There were no consistent changes in plasma insulin levels. The highest rate of growth in these animals is accompanied by an increase in growth hormone concentration followed by an increase in plasma IGF-1. However, despite differences in plasma growth hormone, plasma concentrations of IGF-1 are not different between lines and are not related to between line differences in growth rate.

Animals↗
PubMed1988

Molecular forms of growth hormone in the chicken pituitary gland.

Different forms of GH present in freshly prepared homogenates of chicken pituitary were analysed by high-performance gel permeation chromatography and by immunoblotting following sodium dodecylsulphate polyacrylamide gel electrophoresis, nondenaturing polyacrylamide gel electrophoresis and isoelectric focussing. Chicken GH was found to occur mainly in a monomeric form with a relative molecular mass of 23,500 together with small amounts of interchain disulphide-linked oligomers. No evidence was obtained for proteolytically modified forms of GH nor for a form analogous to the 20K variant of human GH. Isoelectric focussing resolved ten distinct charge variants of chicken GH with isoelectric points between 8.45 and 6.0. The predominant charge variant (pI = 7.85) was present in pituitaries of birds of both sexes at all ages examined (3-114 days) whereas the more acidic forms were not apparent until after day 13. These findings indicate that the chicken pituitary contains different forms of GH, some of which may be developmentally regulated.

Animals↗
PubMed1987

Antitumor activity and pharmacokinetics in mice of 8-carbamoyl-3-methyl-imidazo[5,1-d]-1,2,3,5-tetrazin-4(3H)-one (CCRG 81045; M & B 39831), a novel drug with potential as an alternative to dacarbazine.

A number of 3-alkyl analogues of the experimental antitumor drug mitozolomide [8-carbamoyl-3-(2-chloroethyl)imidazo[5,1-d]-1,2,3,5-tetrazin-4(3H )-one] have been screened against murine tumors in vivo. Only the compounds with a 3-methyl- or 3-bromoethyl group possessed significant antitumor activity against the TLX5 lymphoma. The 3-methyl analogue, 8-carbamoyl-3-methylimidazo[5,1-d]-1,2,3,5-tetrazin-4(3H)-one (CCRG 81045), was investigated further and found to possess good activity, when administered i.p., against the L1210 and P388 leukemias, the M5076 reticulum cell sarcoma, B16 melanoma, and ADJ/PC6A plasmacytoma. The drug was also active when administered p.o. to mice bearing the L1210 leukemia. A daily for 5 days schedule of 100 mg/kg CCRG 81045 produced increases of survival time of treated animals compared to controls of 176 and greater than 235% against the P388 and L1210 leukemias, respectively. In the female C57BL x DBA/2 F1 mouse the 10% lethal dose was 125 mg/kg daily for 5 days. CCRG 81045 was found to undergo mild alkaline hydrolysis and ring fission to form the linear triazene 5-(3-methyltriazen-1-yl)imidazole-4-carboxamide, which is the putative metabolite formed upon metabolic activation of the antitumor drug dacarbazine [5-(3,3-dimethyltriazen-1-yl)imidazole-4-carboxamide]. The half-life of CCRG 81045 at 37 degrees C in 0.2 M phosphate buffer (pH 7.4) was 1.24 h, whereas that of 5-(3-methyltriazen-1-yl)imidazole-4-carboxamide at 25 degrees C was reported to be 8 min (F. H. Shealy and C. A. Krauth, J. Med. Chem., 9:34-37, 1966). The half-life of CCRG 81045 in human plasma in vitro at 37 degrees C was 0.42 h. Pharmacokinetic experiments conducted in BALB/c mice produced plasma profiles of CCRG 81045, administered i.p. or p.o., which showed a rapid absorption phase, elimination half-lives of 1.13 h (i.p.) and 1.29 h (p.o.), and a bioavailability of 0.98.

Animals↗
PubMed1987

Contribution of lipoprotein lipase to differences in fatness between broiler and layer-strain chickens.

The growth of abdominal fat in chickens from broiler and layer-strains up to 10 weeks of age was measured and compared with changes in plasma very low density lipoprotein (VLDL) concentration and tissue lipoprotein lipase activities. The growth of abdominal fat in broilers was much more rapid than in layer-strain chickens. Plasma VLDL concentrations in the two strains were similar up to 5 weeks of age but thereafter concentrations tended to be higher in broilers. Plasma VLDL concentrations in both strains were much lower than those necessary for maximum lipoprotein lipase activity. The lipoprotein lipase activity of abdominal fat increased much more rapidly in broilers than in layer-strain chickens. In both strains the pattern of its increase relative to body weight was similar to that of abdominal fat. Differences in the lipoprotein lipase activity of abdominal fat between strains were attributed to differences in both activity/adipocyte and number of adipocytes. They were reduced or abolished if activity was expressed relative to tissue weight, or to its content of DNA or protein. The results strongly suggest that the greater lipoprotein lipase activity of the abdominal fat pad in broilers is an important factor in its rapid growth.

Adipose Tissue↗
PubMed1987

The dismantling of the mental hospital? Glenside Hospital surveys 1960-1985.

Results of the sixth quinquennial survey of the resident population of Glenside Hospital, Bristol, are reported. The total population continues to fall, but the rate of decline has slowed; the implications of this are discussed. Many patients live in an emotionally impoverished state, friendless and rarely leaving the hospital. Few in-patients are employed, even within the hospital. Considerable provision is made, however, for the employment of day-patients. To effectively resettle and support in the community those currently remaining in hospital will require increasingly extensive provision.

Adult↗
PubMed1987

Renal vasopressin receptors in ageing C57BL/Icrfat mice.

The mechanism of water conservation is impaired in ageing mammals. An age-related defect in the release of vasopressin has been implicated but, more recently, attention has moved to the renal component of the water conservation mechanism. Previous studies using renal cells prepared from mice of different ages have shown that the threshold dose of vasopressin required to elicit a significant rise in cyclic AMP (cAMP) was greater in older animals. The dose-response curve was moved to the right in 35-month-old mice, i.e. the concentration of vasopressin required to give maximum cAMP output was increased. To investigate this further we examined the binding of vasopressin to renal medullary cells maintained in short-term culture, to determine whether the decreased response of cAMP levels to vasopressin is due to changes in hormone-receptor interaction. In 6-month-old male mice the dissociation constant (Kd) was 2.38 nmol/l and the maximum binding of the hormone (Bmax) was 47.6 fmol/10(6) cells, and at 30 months of age Kd was 2.37 nmol/l and Bmax was 47.0 fmol/10(6) cells. In female mice the changes were more complicated because the data for the 6-month-old mice could be split into two groups. It is concluded that there are no age-related differences in the numbers of receptors or their affinity for vasopressin, and that the decreased cAMP response is probably associated with post-receptor mechanisms in this species.

Aging↗
PubMed1986

Cardiovascular haemodynamics and the response of vasopressin, aldosterone, plasma renin activity and plasma catecholamines to head-up tilt in young and old healthy subjects.

Increasing age impairs the regulation of blood pressure during posture change. The neuro-humoral and cardiovascular responses to head-up tilt were analysed in carefully-screened young and healthy elderly individuals. Mean blood pressure was significantly higher in the elderly but there were no differences in total peripheral resistance, heart rate, stroke volume and cardiac index. Age-related interactions were observed in the control of mean blood pressure, heart rate and stroke volume. Total peripheral resistance increased and cardiac index decreased but there was no difference in their control in the young and old. Noradrenaline, vasopressin, plasma renin activity and aldosterone all increased in response to the tilt. These observations indicate differences in the neuroendocrine responses and cardiovascular haemodynamics of young and old healthy individuals to head-up tilt and are particularly important because of all observations were made simultaneously in the same subject. It is suggested that a similar approach should be adopted in the investigation of patients with postural hypotension.

Adult↗
PubMed1985

The effect of age on the control of water conservation in the laboratory mouse--metabolic studies.

Age-related changes in the intake of food and water, and the output of faeces and urine were investigated in C57BL/Icrfat mice of 6 and 24 months of age. Animals were singly housed in a metabolic cage for a period of 30 days. 14 days were allowed for acclimatization before the animals were dehydrated for 24 hours. 10 days of rehydration were allowed prior to a hyperosmotic challenge with 3% sodium chloride in the drinking water. The animals were then observed for 5 more days of rehydration. Urine was collected and analysed with regard to sodium, potassium, urea and vasopressin output/24 hours (/100g body weight), and the osmotic pressure of the urine was determined. Data were analysed by a 2 factor analysis of variance with repeated measures on one factor. Significant changes were detected in the control of body weight, potassium, sodium and urea outputs. No age-differences were detected in the intake of food or water, the output of faeces or urine, the urine osmotic pressure or the excretion of vasopressin. However, significant changes in these variables were detected in both age groups on the days of physiological challenge. The conclusion drawn is that in the mouse strain studied, and for the period of the lifespan investigated, there is no age related defect in the secretion of vasopressin. However, there are trends in the data suggesting a decreased responsiveness of the kidney with age.

Age Factors↗
PubMed1985

Antitumour imidazotetrazines, Part IX. The pharmacokinetics of mitozolomide in mice.

Mitozolomide is a novel antitumour agent showing a broad spectrum of activity against murine tumours and is currently undergoing Phase I clinical evaluation in the UK. We have conducted an animal pharmacokinetic study using male BALB/c mice as a pre-requisite to the clinical work. Mice were dosed i.p. at 5 dose levels (0.25-20 mg kg-1) and the oral and transdermal routes of administration were investigated at 20 mg kg-1. The analytical data produced a good fit to a simple open one-compartment pharmacokinetic model with an elimination half-life of the drug from plasma of between 0.68 and 0.88 h over the 0.25-20 mg kg-1 range covered. There was no evident dose dependency over this range and studies with two formulations showed mitozolomide to have good systemic availability when administered via the oral route (F values of 0.66 and 0.81). The drug was also found to be systemically available when administered topically in dimethylsulfoxide (F = 0.47). Mitozolomide shows many biochemical and biological similarities to the clinically used nitrosoureas BCNU and CCNU but our results show that it differs markedly in its kinetics from these two agents, with mitozolomide having relatively sustained plasma levels. It is hoped that this may be of therapeutic benefit if these levels are reflected in relative tumour concentrations.

Administration, Oral↗
PubMed1984

Plasma arginine vasopressin in dehydrated elderly patients.

Plasma arginine vasopressin was assessed by radioimmunoassay in dehydrated elderly patients and healthy young and elderly control subjects. The elderly control subjects' AVP levels were higher than the young control subjects (Kruskal-Wallis rank sums z = 3.29; P less than 0.001). Despite similar plasma osmolalities, after correction for the osmotic contribution of urea, the patients' AVP levels were higher than the elderly controls (z = 4.16; P less than 0.001) and were in the range reported to evoke a maximal antidiuretic response. It is concluded that the dehydration which often accompanies acute illness in the elderly is not primarily a consequence of inadequate AVP release.

Adult↗
PubMed1984

Effect of ageing on cyclic AMP output by renal medullary cells in response to arginine vasopressin in vitro in C57BL/Icrfat mice.

The effect of age on the cyclic AMP (cAMP) response to increases in the concentration of arginine vasopressin in the presence of isobutyl methylxanthine (100 mumol/l) was studied in an in-vitro renal cell suspension prepared from C57BL/Icrfat mice at 6, 12, 18, 24, 29 and 35 months of age. Comparison of the response of the preparation to vasopressin, calcitonin and parathyroid hormone suggested that it was enriched with renal medullary cells. Basal cAMP output was similar throughout but the threshold dose of vasopressin increased from 1 X 10(-11) mol/l (6, 12 and 18 months of age) to 1 X 10(-10) mol/l (24, 29 and 35 months of age). The dose-response curve in 35-month-old mice was shifted to the right with the concentration of vasopressin required to give half maximal cAMP increased from 9.4 +/- 0.37 X 10(-11) mol/l (6 months) to 3.5 +/- 1.6 X 10(-10) mol/l (35 months). Maximum cAMP output at 1 X 10(-9) mol/l was also reduced in the same animals (stimulated:basal ratio, 51.22 +/- 19.12 at 6 months; 11.50 +/- 6.02 at 35 months). The results suggest that the lack of renal response to vasopressin in terms of cAMP metabolism may play a role in the well-documented age-related decline in urine-concentrating ability in experimental animals and elderly people.

Aging↗
PubMed1983

Polyuria and inappropriate secretion of arginine vasopressin in hypothalamic sarcoidosis.

Inappropriate arginine vasopressin release and polyuria with excessive thirst were found in a patient with hypothalamic sarcoidosis subsequently confirmed at autopsy. He became intensely thirsty during a 5% saline infusion at a plasma osmolality of 274 mosmol/liter. The normal thirst threshold under these conditions is 294 +/- 3 mosmol/liter (+/- SD). An increase in radioimmunoassayable arginine vasopressin was detected at an inappropriately low plasma osmolality, and free water clearance was negative despite a plasma osmolality of 265 +/- 5 mosmol/liter with ad libitum fluid intake. The syndrome of inappropriate antidiuresis has not been described previously in combination with hypothalamic sarcoidosis. Demeclocycline therapy was associated with an exacerbation of the patient's polyuria. Propranolol administration, however, was associated with a reduction of urine output and an increase in plasma osmolality to 279 mosmol/liter on one and 299 mosmol/liter on another occasion.

Adult↗
PubMed1982

Quantitation of muscle function in children: a prospective study in Duchenne muscular dystrophy.

A protocol has been developed for the quantitative assessment of muscle function in children with muscle disease. It includes total muscle strength (% MRC) based on a clinical assessment of strength of 32 groups using the 6-point MRC grading; the force of 8 selected muscle groups measured with a specially designed electromyometer; a motor ability score based on 20 consecutive motor activities; walking times over 28 and 150 feet, and recording of muscle contractures. A 3-year sequential study of 61 boys with Duchenne dystrophy showed progressive decline of muscle strength with age, a close correlation of total strength and the motor ability score (r = 0.89), and a curvilinear relationship of muscle strength with walking times over 28 and 150 feet (r = 0.78 and 0.79, respectively). A profile of the natural progression of Duchenne dystrophy has been established which could serve as a reference base for the assessment of cases at varying ages and their response to therapy and management.

Aging↗