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Biomedical subjects

J Pagan

Publications and source records attributed to J Pagan.

17 recordsLinked to original sources
PubMed2002

Comparison of the metabolic responses of trained Arabians and Thoroughbreds during high- and low-intensity exercise.

In order to compare the metabolic responses to exercise in 2 similarly managed breeds of horses, 5 Arabian (AR) and 5 Thoroughbred (TB) horses, fed an identical diet with a similar diet and exercise training programme for the 2 preceding months, undertook 3 treadmill (3 degree incline) trials with a minimum of 7 days between tests: 1) an incremental test (MAX) for determination of aerobic capacity, V(LA4) and lactate threshold (LT; the percentage of VO2max when plasma lactate = 4 mmol/l); 2) a single high-speed exercise test (SPR) at 115% VO2max for estimation of maximal accumulated oxygen deficit (MAOD) and 3) a 90 min test at 35% VO2max (LO). VO2max (P<0.001) and running speed (P<0.05) at VO2max were higher in TB (mean +/- s.e. 154 +/- 3 ml/kg/min at 12.9 +/- 0.5 m/s) than in AR (129 +/- 2.5 ml/kg/min at 11.8 +/- 0.2 m/s). Total run time during MAX was greater (P<0.05) in TB (10.5 +/- 0.5 min) than in AR (93 +/- 0.3 min). However, V(LA4) and LT were not different between groups. Run time during SPR (TB 149 +/- 16; AR 109 +/- 11 s) and MAOD (TB 88 +/- 4; AR 70 +/- 6 ml O2/kg) were higher (P<0.05) in the TB group. During LO, FFA were higher (P<0.05) and the respiratory exchange ratio (RER) lower (P<0.05) in AR than in TB between 60 and 90 min, of exercise, indicating a greater use of fat for energy. These metabolic differences may reflect breed variation in muscle fibre types. However, further studies are needed to determine the mechanisms underlying the apparent breed differences in energy metabolism during exercise.

Animals↗
PubMed2000

Mortality and paraplegia after thoracoabdominal aortic aneurysm repair: a risk factor analysis.

BACKGROUND: Recent recommendations regarding thoracoabdominal aortic aneurysm (TAAA) management have emphasized individualized treatment based on balancing a patient's calculated risk of rupture with their anticipated risk of postoperative death or paraplegia. The purpose of this study was to enhance this risk-benefit decision by providing contemporary results and determining which preoperative risk factors currently predict mortality and paraplegia after TAAA surgery. METHODS: Risk factor analyses based on data regarding 1,220 consecutive patients undergoing TAAA repair from 1986 through 1998 were performed using multiple logistic regression with step-wise model selection. RESULTS: The 30-day mortality rate was 4.8% (58 of 1,220) and the incidence of paraplegia was 4.6% (56 of 1,206). For elective cases, predictors of operative mortality included renal insufficiency (p = 0.0001), increasing age (p = 0.0005), symptomatic aneurysms (p = 0.0059), and extent II aneurysms (p = 0.0054). Extent II aneurysms (p = 0.0023) and diabetes (p = 0.0402) were predictors of paraplegia. CONCLUSIONS: These risk models may assist in decisions regarding elective TAAA operations. For patients who are acceptable candidates, contemporary surgical management provides favorable results.

Adolescent↗
PubMed2000

Six years' experience of prophylactic oral vitamin K.

AIMS: The ability of oral vitamin K to eliminate all risk of vitamin K deficiency bleeding during the first three months of life was studied. METHODS: Babies (n=182,000) in the north of England judged well enough to be offered milk within 12 hours of birth were given 1 mg of phytomenadione (vitamin K(1)) suspended in a medium chain triglyceride oil by mouth at delivery between 1993 and 1998. The parents of those who were breastfed were given a further three doses to give to the baby once every two weeks after discharge. RESULTS: Four breastfed babies developed late vitamin K deficiency bleeding. In two, staff failed to follow policy guidelines, and in two there was undiagnosed alpha(1) antitrypsin deficiency. Audit suggested that 93% of breastfed babies had all four doses, as advised. CONCLUSIONS: An oral product that parents can administer themselves would be popular if licensed, but the total dose offered may need to be more than in this study if babies with undiagnosed liver disease are to be protected.

Administration, Oral↗
PubMed1995

Mutagenesis and reversion analysis of residue Met-209 of the beta-subunit of Escherichia coli ATP synthase.

Residue beta-Met-209 is conserved in all known F1-ATPase sequences, and the mutation beta M209I in Escherichia coli causes profound inhibition of ATP synthesis and hydrolysis. Based on the properties of this mutant it had previously been proposed that residue beta-209 lies close to the site of catalysis. Two approaches were used to study this residue further. First, revertants were sought. Only wild-type and beta-Ser-209 were found; the Ser revertants involved a two-base change. Significantly, Ser is found at the equivalent position in the homologous vacuolar and archaebacterial ATPases. Second, all 20 natural amino acids were placed at position beta-209 by mutagenesis, and catalytic properties of the mutants were analyzed. The results showed that only a limited set of residues supported significant growth or ATPase activity, and that many of the mutations impacted severely on catalysis. X-ray structure analysis of the bovine enzyme has revealed that residue beta-Met-209 lies only 3.1. A from residue beta-Glu-181, which has been proposed to act as catalytic base. The results reported here emphasize that, in this discrete region of the catalytic site, specific stereo-chemical constraints on structure are critical for catalysis.

Adenosine Triphosphate↗
PubMed1991

Characterization of Escherichia coli ATP synthase beta-subunit mutations using a chromosomal deletion strain.

(1) We constructed Escherichia coli strain JP17 with a deletion in the ATP synthase beta-subunit gene. JP17 is completely deficient in ATP synthase activity and expresses no beta-subunit. Expression of normal beta-subunit from a plasmid restores haploid levels of ATP synthase in membranes. JP17 was shown to be efficacious for studies of beta-subunit mutations. Site-directed mutants were studied directly in JP17. Randomly generated chromosomal mutants were identified by PCR and DNA sequencing, cloned, and expressed in JP17. (2) Eight novel mutations occurring within the putative catalytic nucleotide-binding domain were characterized with respect to their effects on catalysis and structure. The mutations beta C137S, beta G152D, beta G152R, beta E161Q, beta E161R, and beta G251D each impaired catalysis without affecting enzyme assembly or oligomeric structure and are of interest for future studies of catalytic mechanism. The mutations beta D301V and beta D302V, involving strongly conserved carboxyl residues, caused oligomeric instability of F1. However, growth characteristics of these mutants suggested that neither carboxyl side chain is critical for catalysis. (3) The mutations beta R398C and beta R398W rendered ATP synthase resistant to aurovertin, giving strong support to the view that beta R398 is a key residue in the aurovertin-binding site. Neither beta R398C or beta R398W impaired catalysis significantly.

Aurovertins↗
PubMed1990

Tight ATP and ADP binding in the noncatalytic sites of Escherichia coli F1-ATPase is not affected by mutation of bulky residues in the 'glycine-rich loop'.

It is shown that ATP dissociates very slowly (koff less than 6.4 x 10(5) s-1, t1/2 greater than 3 h) from the three noncatalytic sites of E. coli F1-ATPase and that ADP dissociates from these three sites in a homogeneous fashion with koff = 1.5 x 10(-4) s-1 (t1/2 = 1.35 h). Mutagenesis of alpha-subunit residues R171 and Q172 in the 'glycine-rich loop' (Homology A) consensus region of the noncatalytic sites was carried out to test the hypothesis that unusually bulky residues at these positions are responsible wholly or partly for the observed tight binding of adenine nucleotides. The mutations alpha Q172G or alpha R171S,Q172G had no effects on ATP or ADP binding to or rates of dissociation from F1 noncatalytic sites. KdATP and KdADP of isolated alpha-subunit were weakened by approximately 1 order of magnitude in both mutants. The results suggest that neither residue alpha R171 nor alpha Q172 interacts directly with bound nucleotide, and show that the presence of bulky residues per se in the glycine-rich loop region of F1-alpha-subunit is not responsible for tight binding in the noncatalytic sites.

Adenosine Diphosphate↗
PubMed1990

Mutations in alpha-subunit of Escherichia coli F1-ATPase obtained by hydroxylamine-mutagenesis of plasmids carrying the uncA gene.

In order to generate mutants randomly in the Escherichia coli uncA gene (encoding the alpha-subunit of F1-ATPase), plasmids carrying uncA were treated in vitro with hydroxylamine. Restriction fragments of the mutated uncA gene were then reconstructed into plasmid pDP34, which expresses all of the F1F0 structural genes, and the reconstructed mutant plasmids were expressed in a strain carrying a deletion of chromosomal uncA. Each of the mutations was characterized by DNA sequencing, growth assays, and biochemical assays of membrane preparations. Three nonsense and one frameshift mutation were identified and their properties were studied briefly. Eight new missense mutations were identified and characterization of their properties is described. These eight mutations were R139H, A177V, R210C, R303C, A306V, T343I, G351S, and P370L.

Amino Acid Sequence↗
PubMed1989

Identification of a mutation in Escherichia coli F1-ATPase beta-subunit conferring resistance to aurovertin.

A mutation conferring aurovertin resistance on Escherichia coli F1-ATPase was identified as R398----H in the F1 beta-subunit. Beta-subunit from the mutant does not bind aurovertin; therefore our results suggest the region of sequence around residue beta-398 is involved in aurovertin binding. Since nucleotide and aurovertin binding to isolated beta-subunit are not mutually exclusive, the data further suggest that the beta-subunit catalytic nucleotide-binding domain does not include residue 398. The mutation prevented aurovertin inhibition of ATPase at pH 6 and 8.5, implying charge on the arginine side-chain is not a major determinant of aurovertin binding or that the pK of R398 is shifted due to a peculiar environment. The equivalent residue is usually arginine in F1 beta-subunits of different species; notably in the aurovertin-insensitive thermophilic bacterium PS3 F1-ATPase, this residue is phenylalanine.

Aurovertins↗
PubMed1988

Directed mutagenesis of the strongly conserved lysine 175 in the proposed nucleotide-binding domain of alpha-subunit from Escherichia coli F1-ATPase.

The alpha-subunit of Escherichia coli F1-ATPase contains an adenine-specific noncatalytic nucleotide-binding domain. A recent proposal (Maggio, M. B., Pagan, J., Parsonage, D., Hatch, L., and Senior, A. E. (1987) J. Biol. Chem. 262, 8981-8984) suggested that this domain is formed by residues 160-340, approximately, in alpha-subunit. Within this proposed domain is a sequence Gly-X-X-X-X-Gly-Lys which is conserved in a large and diverse group of nucleotide-binding proteins and is thought to interact with phosphate groups of bound nucleotide. In this work, residue alpha Lys-175, the terminal residue of the above conserved sequence in F1-alpha-subunit, was mutagenized to Ile or Glu. The specific activity of purified mutant F1-ATPase was reduced by 2.5-fold (Ile) or 3-fold (Glu). Apparent binding of ATP to alpha-subunit, as measured by the centrifuge column procedure, was strongly impaired and ATP-induced conformational change in alpha-subunit, as measured by protection against trypsin proteolysis, was nearly abolished in both mutants. The results suggest that residue alpha Lys-175 is located within the nucleotide-binding domain of alpha-subunit, and that this residue is functionally involved in nucleotide binding. The results support previous suggestions that the alpha-subunit nucleotide-binding site is not involved, directly or indirectly, in catalysis.

Binding Sites↗
PubMed1987

The defective proton-ATPase of uncA mutants of Escherichia coli. Identification by DNA sequencing of residues in the alpha-subunit which are essential for catalysis or normal assembly.

A group of mutant uncA alleles, affecting essential residues of the alpha-subunit of Escherichia coli proton-ATPase, have been identified by intragenic complementation mapping, cloning, and DNA sequencing. One of the mutations, uncA450, abolishes normal assembly of F1-ATPase. The amino acid substitution found was Glu-299----Lys, which is predicted to lie in an alpha-helix in alpha-subunit. The reversal of the charge at residue 299 is a likely cause of defective assembly. The uncA462 allele causes impairment of catalysis while allowing normal assembly of membrane-bound F1-ATPase. The amino acid substitution found was Ser-347----Phe. Three mutations which impair catalysis but do not cause structural perturbation of either membrane-bound or solubilized F1ATPase were characterized as follows: uncA401, Ser-373----Phe; uncA447, Gly-351----Asp; uncA453, Ser-375----Phe. We predict here that the nucleotide-binding domain of alpha-subunit is formed by the amino acids in the sequence from residue 160 to approximately residue 340. The mutations which cause impairment of catalysis lie in a short segment between residues 347-375 of alpha-subunit, at the C-terminal end of the predicted nucleotide-binding domain. This segment is suggested to be important for beta-alpha-beta intersubunit conformational interaction involved in positive catalytic cooperativity in F1-ATPase.

Alleles↗
PubMed1984

Efficacy and tolerability of dibenzepin administered by intravenous drip infusion to severely depressed patients.

46 severely depressed inpatients received 10-hour intravenous infusions supplying a dose of 720 mg dibenzepin daily on 2 or some cases 3 successive days. The patients were kept under close psychiatric surveillance and their cardiovascular function was carefully monitored. The shorter infusions were as effective as infusions of the same dose of Noveril over 48 h, and did not lead to a greater frequency or severity of general or cardiovascular side effects. The 10-hour infusion regimen thus represents a new mode of treatment for severe depression which is less demanding both technically and in terms of nursing care.

Administration, Oral↗