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Biomedical subjects

P Holloway

Publications and source records attributed to P Holloway.

13 recordsLinked to original sources
PubMed1998

Alteration of the substrate range of haloalkane dehalogenase by site-directed mutagenesis.

We attempted to expand the range of chlorinated solvents degraded by Xanthobacter autotrophicus GJ10 to include trichloroethylene by the rational modification of the enzyme haloalkane dehalogenase. The amino acids Phe164, Asp170, Phe172 and Trp175 were individually replaced with alanine by site-directed mutagenesis. All substitutions produced enzymes with lower than wild type activity with 1,2-dichloroethane. The Phe164Ala and Asp170Ala mutants were 3 and 2 times more active than was the wild type enzyme in dechlorinating 1,6-dichlorohexane. The Asp170Ala mutant resembled the wild type enzyme in its relative activity against longer chain substrates. No mutant was active with trichloroethylene.

Alanine↗
PubMed1997

Predicting which adult patients will need treatment over the next year.

This prospective study was conducted to determine factors important in predicting which regularly attending adult patients would receive first, restorations or extractions for any reason (receiving treatment) and, second, restorations or extractions undertaken specifically for caries (receiving treatment related to caries). Baseline and incremental clinical data were obtained from 24 general dental practitioners on a group of their regularly attending, dentate adult patients over a 12-month period. The patients completed a postal questionnaire with questions relating to dental health behaviour, attitudes, knowledge, and social factors. Complete data were obtained from 2553 patients. Thirty-one variables were identified as potential predictors for the two dependent variables receiving treatment and receiving treatment related to caries, and logistic regression models were fitted. Receiving treatment was associated with having fewer sound teeth and more anterior fillings, posterior fillings and crowns (P < 0.001). The dentist's prediction of the need for treatment related to caries and the patient's own prediction of the need for a filling were also important in the model (P < 0.001). Some of these variables, together with having received recent medical treatment and taking sugar in tea or coffee were also found to predict treatment related to caries. The model for receiving treatment related to caries was more successful at predicting the patient's individual risk but the model for receiving treatment was slightly better at classifying patients into whether or not they received treatment. It is reassuring that the common assumptions made by the dental practitioners of their patient's risk have received statistical validation.

Adult↗
PubMed1996

Identification and analysis of the dissimilatory nitrous oxide reduction genes, nosRZDFY, of Rhizobium meliloti.

The complete nos region essential for dissimilatory nitrous oxide reduction by the endosymbiotic diazotroph Rhizobium meliloti was identified in a cosmid (pYC7) carrying a 10.1-kb EcoRI fragment of the nod megaplasmid. This gene region was localized by Southern hybridization and Tn5 mutagenesis to within 8 kb downstream from the fixGHIS cluster. Nucleotide sequence determination of a 4.6-kb DNA segment including the structural gene nosZ and its flanking regions showed sequence homology and similarity in genetic organization with the nosRZDFY genes of Pseudomonas stutzeri Zobell. The genes were arranged in three complementation groups, comprising the nosZ structural gene, the nosR regulatory gene, and the nosDFY copper-processing genes. The derived amino acid sequence of the R. meliloti nosZ product (a multi-copper nitrous oxide reductase) was more similar to those of the analogous gene products of Paracoccus and Pseudomonas species than to that of Alcaligenes eutrophus. The nosZ gene was preceded by nosR, which encodes a regulatory protein containing C-terminal cysteine clusters similar to those present in the 4Fe-4S binding region of bacterial ferredoxins, The nosDFY genes, located downstream from nosZ, were identified as copper-processing genes encoding a periplasmic protein, an ATP/GTP-binding protein, and a membrane protein presumably forming a copper-processing system. A consensus sequence for an Anr- or Fnr-binding site similar to that in the upstream sequence of nosZ in Paracoccus denitrificans or P. stutzeri was absent in R. meliloti. No rpoN-binding site preceding the nos genes was detected, and none of the Tn5 insertions in the nos gene region affected symbiotic N2-fixing ability.

Amino Acid Sequence↗
PubMed1994

The nucleotide sequence and initial characterization of pyruvate decarboxylase from the yeast Hanseniaspora uvarum.

We have isolated a pyruvate decarboxylase (PDC) gene from the yeast Hanseniaspora uvarum using the Saccharomyces cerevisiae PDC1 gene as a probe. The nucleotide sequence of this gene was determined and compared to PDC genes from yeast and other organisms. The H. uvarum PDC gene is more than 70% identical to the S. cerevisiae PDC isozymes and possesses a putative thiamine diphosphate binding site. The PDC enzyme was purified and partially characterized. The H. uvarum PDC was very similar to other known PDCs; the Km for pyruvate was 0.75 mM, and the enzyme is a homotetramer with subunits of M(r) = 57,000.

Amino Acid Sequence↗
PubMed1993

Plasmodium falciparum: in vitro studies of the pharmacodynamic properties of drugs used for the treatment of severe malaria.

The speed and stage specificity of antimalarial drug action on the metabolic activities of cultured Plasmodium falciparum were studied for chloroquine (CQ), quinine (QN), artemisinin (AR), and sodium artelinate (SA). CQ had the most rapid onset of action on [3H]hypoxanthine and [3H]isoleucine uptake, reaching 50% of its maximum effect in 1.8 hr compared with 3.5-7.4 hr for the other three drugs. In contrast there was a lag time of 1-4 hr before AR and SA had a measurable inhibitory effect, although after this delay antimalarial action was very rapid. Parasite glycolysis was relatively drug resistant; the inhibition of lactate production was < 60% of that for [3H]hypoxanthine and [3H]isoleucine uptake. The susceptibility of P. falciparum changed markedly as the parasite matured. Maximum drug effects occurred at the late ring and early trophozoite stage, which corresponds to the time at which the most rapid increases in synthetic and glycolytic activities occur. Mature schizonts and young rings were relatively unaffected by the antimalarial drugs. Young rings were particularly resistant to QN. Schizonts multiplied successfully in the presence of relatively high concentrations of all four drugs. The two artemisinin compounds had the broadest time window of action and may be particularly suitable for the treatment of severe malaria.

Animals↗
PubMed1993

The isolation and nucleotide sequence of the pyruvate decarboxylase gene from Kluyveromyces marxianus.

We have determined the nucleotide sequence of a 2360-basepair (bp) region of the Kluyveromyces marxianus genome containing the structural gene for the enzyme pyruvate decarboxylase (PDC). Comparison of the deduced amino-acid sequence of this gene to that of the Saccharomyces cerevisiae PDC genes reveals extensive homology including a motif common to thiamin diphosphate-dependent enzymes.

Amino Acid Sequence↗
PubMed1992

Absorption of intramuscular phenobarbitone in children with severe falciparum malaria.

The absorption of intramuscular phenobarbitone 7 mg.kg-1 was studied in 11 Karen children aged between 1.7 and 11 y with severe falciparum malaria. Eight of the children were comatose. Clinical findings were compared with those in 9 further children with severe malaria of similar age range (four of whom were unconscious), who received an identical placebo. One child, who had received placebo, had repeated convulsions and died 1 h after admission to hospital. The remainder made an uncomplicated recovery. There were no convulsions subsequent to treatment, although the study was too small to assess anticonvulsant efficacy. There was no observable toxicity, but phenobarbitone recipients had a significant tendency to deepen in their level of coma or to become sleepy within the 4 h after drug administration. Phenobarbitone was rapidly absorbed, reaching a mean (range) peak concentration of 34.2 [29.3-42.6] mumol.l-1 in a median (range) of 4 (2.5-12) h. These values are comparable to those previously reported in healthy children and in children with febrile convulsions. Intramuscular phenobarbitone is well absorbed in children with severe malaria; the optimum prophylactic anticonvulsant dose remains to be determined.

Absorption↗
PubMed1991

Calcium and phosphate metabolism in acute falciparum malaria.

1. Mineral homeostasis was investigated in 172 Thai adults with acute falciparum malaria at presentation (87 males, 85 females; mean age 30 years), and prospectively in a subgroup of 10 severely ill patients. 2. Mild, asymptomatic hypocalcaemia (corrected plasma calcium concentration 1.79-2.11 mmol/l) was found in 61 cross-sectional study patients (35.5%), with no difference between those with uncomplicated (2.16 +/- 0.10 mmol/l, mean +/- SD, n = 89) and severe (2.18 +/- 0.15 mmol/l, n = 83, P = 0.36) infections. Six prospectively studied patients were hypocalcaemic during treatment; simultaneous serum intact parathormone concentrations were inappropriately low (less than 5.0 pmol/l), but rose in three patients to high levels (11.8-16.4 pmol/l) on the fifth day. 3. Plasma phosphate concentration was decreased (less than 0.80 mmol/l) on admission in 74 patients (43.0%) and increased (greater than 1.45 mmol/l) in 15 (8.7%). Severe phosphate depletion (plasma phosphate concentration less than 0.30 mmol/l) occurred in 14 patients, of whom 11 had severe infections. Serum phosphate concentrations in the prospective study patients on admission (0.59 +/- 0.23 mmol/l) correlated significantly with the simultaneous renal threshold phosphate concentration (0.68 +/- 0.33 mmol/l; r = 0.607, P less than 0.025) and both parameters rose in parallel during treatment. 4. Plasma magnesium concentrations were normal (0.75-1.05 mmol/l) in 108 patients (62.8%); 45 cases (26.1%) had hypermagnesaemia and 19 (11.0%) had hypomagnesaemia. 5. These data suggest that mild hypocalcaemia is common in malaria regardless of disease severity; a depressed parathormone response may contribute. Despite malaria-associated haemolysis, hypophosphataemia is also common, but can be severe.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute Disease↗
PubMed1986

Magnetic resonance spectroscopy for the determination of renal metabolic rate in vivo.

The method of magnetic resonance spectroscopy has been validated and applied to the determination of renal metabolic rate in vivo. Using an indwelling detector coil, 31P NMR spectra from one kidney of anesthetized rats were quantified. The concentration of ATP was the same as that determined enzymatically, but both ADP and Pi were substantially lower. Only 25% of renal Pi and virtually none of the ADP were detected by NMR. The remainder is assumed to be bound to proteins. These concentrations of metabolites contributed to a significantly increased phosphorylation potential, which in turn should increase the free energy of hydrolysis of ATP. Saturation transfer, a non-destructive magnetic technique for the measurement of chemical exchange, was readily able to detect synthesis of ATP from ADP and inorganic phosphate. The rate of ATP synthesis determined was comparable to that determined in parallel studies of renal oxygen consumption. An ATP:O ratio of approximately 2 was found, indicating that fatty acid is the preferred fuel of respiration of the rat kidney in vivo.

Adenosine Diphosphate↗
PubMedDate not supplied

Communication: the key to avoiding query and reporting standards violations.

Without carefully designed communication protocols, hospitals can fail to meet the query and reporting requirements imposed by the Joint Commission on Accreditation of Healthcare Organizations, the National Practitioner Data Bank, and state medical societies. This article tracks the history of the requirements, outlines current standards, and offers specific recommendations for operational procedures to avoid potential pitfalls.

Accreditation↗
PubMedDate not supplied

Early metabolic effects of hypotension on rat kidney.

Saturation-transfer phosphorus nuclear magnetic resonance (STNMR) has been applied to the rat kidney in vivo. The rate of renal metabolism determined by this method compares favorably with the renal oxygen consumption, assuming an ATP:oxygen ratio of 2. Hemorrhagic hypotension resulted in a 20% fall in renal blood flow and a significant fall in oxygen consumption. The rate of renal metabolism fell by 50%. This rate of ATP synthesis was below that required to maintain a normal [ATP], but renal [Pi] was not increased. When renal perfusion was reduced by 60%, intrarenal [Pi] rose. When [P1] was elevated, the method os STNMR no longer gave a reliable measure of the rate ATP synthesis, indicating that this new Pi pool was not in rapid chemical exchange with ATP. STNMR represents a useful noninvasive means of monitoring renal metabolic rate, with limitations due to insensitivity and the existence of multiple pools of intrarenal Pi.

Adenosine Diphosphate↗