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Biomedical subjects

A F Brodie

Publications and source records attributed to A F Brodie.

At least 19 recordsLinked to original sources
PubMed1995

Management of seizures in the elderly: a survey of UK geriatricians.

The incidence and prevalence of epilepsy increase substantially with old age. Despite this, the investigation and management of this patient population remains a grey area. Four hundred and eleven (53%) consultant geriatricians responded to a questionnaire exploring their approach to seizures in the elderly in order to establish an overview of current clinical practice. Between one and five patients presenting with seizures, predominantly aged between 75-85 years, were reviewed monthly. Seventy per cent of geriatricians undertook to investigate the patients themselves with biochemical and haematological profiles performed by most. Electroencephalography and computerized tomographic scanning were routinely requested by a quarter of responders. Only 58% would themselves initiate therapy with antiepileptic drugs, with 16% of consultants starting treatment following the first seizure, 59% after a second and 5% after a third. Phenytoin was first choice for generalized tonic-clonic seizures with carbamazepine preferred for partial seizures. If good control was not obtained, 67% would substitute another first line drug, while 27% would add in a second. Less than 3% would use the new anticonvulsants lamotrigine or vigabatrin. Sixty per cent monitored anticonvulsant concentrations in patients with poor control or suspected toxicity. A wide variability was seen in the current approach to seizures in the elderly, which reflects a lack of established practice. Epilepsy clinics for the elderly would encourage structured research into the many unanswered questions affecting the care of older people with seizures.

Aged↗
PubMed1994

An outcome audit at the epilepsy clinic: results from 1000 consecutive referrals.

The clinical details, specialist investigation, pharmacological treatment and outcome in 1000 consecutive patients referred to the epilepsy clinic at the Western Infirmary in Glasgow were reviewed. Data were collected by detailed proforma at initial referral and from the clinical notes at audit. 524 (52%) of the patients were 30 years of age or younger. 527 (53%) were sent to the clinic by their general practitioner with the majority of the remainder being referred by another hospital consultant. 240 (24%) came from outwith greater Glasgow. In 253 (25%) patients a possible predisposing factor was identified, usually a major head injury or alcohol abuse. Sixty (6%) patients suffered from additional mental handicap. Focal spike and wave activity was identified in 26% of electroencephalograms. One or more abnormality was found in 33% of the 356 patients who underwent computerized tomographic brain scanning. In 13 (11%) of these, a tumour was demonstrated. On referral, 65% of patients were being treated with antiepileptic drugs; 58% were on monotherapy and 42% took more than one anticonvulsant. The figure at audit was 77% treated, with a higher proportion (68%) than at referral receiving monotherapy (P < 0.005). Sedative anticonvulsants, such as phenobarbitone and primidone, were withdrawn in 29% of cases. 50% of the 519 patients, for whom seizure frequency data were available for the years before referral and audit, were seizure-free and seizure numbers had been reduced by 50% or more in a further 23%. A poorer response to treatment was associated with mental handicap (P < 0.05), birth injury (P < 0.01), and partial or secondary generalized seizures (P < 0.005).

Adolescent↗
PubMed1993

Five years of anticonvulsant monitoring on site at the epilepsy clinic.

The influence of antiepileptic drug monitoring on site at the epilepsy clinic was assessed in a prospective way by recording physicians' decisions before and after the drug concentration became available over the five-year period including 1986 to 1990. A total of 2,857 assays (1,121 carbamazepine, 798 phenytoin, 622 sodium valproate, and 72 phenobarbital) were performed during 2,696 hospital attendances on 618 epileptic patients. A gradual reduction in the proportion of visits monitored annually was observed (1986: 42%; 1987: 51%; 1988: 44%; 1989: 39%; 1990: 28%; p < 0.01). In 481 (17.8%) visits a change in management resulted from knowledge of the drug concentration. This was not an automatic response, as a large minority of unexpected results did not produce such an alteration. The proportion of decisions to adjust the drug dose after an unexpected result, however, tended to increase over the years of the study (1986: 43%; 1987: 70%; 1988: 60%; 1989: 78%; 1990: 54%; p < 0.01). Changes in anticonvulsant dose following an unexpected result were observed in "low" (59%), "therapeutic" (61%), and "high" (52%) concentration categories. Possible benefit for patients can be inferred from the observation that more assay results fell within the target range in the last (61%) compared with the first (33%) year of monitoring (p < 0.01).

Anticonvulsants↗
PubMed1981

Interaction of vanadate with membrane-bound ATPase from Mycobacterium phlei.

Vanadate inhibited the formation of proton gradient and membrane potential as well as Ca2+ transport by everted membrane vesicles from Mycobacterium phlei, with half-maximal inhibition occurring at 5 to 14 microM. That this is due to the inhibition of the proton-translocating ATPase was suggested by the observation that the inhibition described above occurred only when the processes were driven by the hydrolysis of ATP but not when energized by the oxidation of succinate and NADH. Furthermore, vanadate did indeed inhibit ATP hydrolysis by these membrane vesicles. Although the inhibition of ATP hydrolysis could be demonstrated only in the presence of high concentrations (e.g. 11 mM) of Mg2+, this was presumably due to the fact that we were measuring the sum of ATP hydrolysis by both coupled and partially uncoupled enzymes. This is the first reported effect of vanadate on bacterial proton-translocating ATPase.

Adenosine Triphosphatases↗
PubMed1981

Active transport of Ca2+ in bacteria: bioenergetics and function.

The bioenergetics of Ca2+ transport in bacteria are discussed with special emphasis on the interrelationship between transport and other cellular functions such as substrate oxidation by the respiratory chain and oxidative phosphorylation. The unusual polarity of Ca2+ movement provides an exceptional tool to compare active transport and other ATP requiring or generating processes since this ion is actively taken up by everted vesicles in which the coupling-factor ATPase is exposed to the external medium. As inferred from studies with everted vesicles, the active extrusion of Ca2+ by whole cells can be accomplished by substrate driven respiration, hydrolysis of ATP or as in the case of Streptococcus faecalis by a nonhydrolytic unknown process which involves ATP directly. Substrate oxidation and the hydrolysis of ATP result in the generation of a pH gradient which can energize the Ca2+ uptake directly (Ca2+/H+ antiport) or via a secondary Na+ gradient (Ca2+/Na+ antiport). In contrast to exponentially growing cells sporulating Bacilli accumulate Ca2+ during the synthesis of dipicolinic acid. Studies involving Ca2+ transport provided evidence in support of the hypothesis that the Mg2+ ATPase from Escherichia coli not only provides the driving force for various cellular functions but exerts a regulatory role by controlling the permeability of the membrane to protons. The different specificity requirements of various naphthoquinone analogs in the restoration of transport or oxidative phosphorylation, after the natural menaquinone has been destroyed by irradiation, has indicated that a protonmotive force is sufficient to drive active transport. However, in addition to the driving force (protonmotive force) necessary to establish oxidative phosphorylation, a specific spatial orientation of the respiratory components, such as the naphthoquinones, is essential for the utilization of the proton gradient or membrane potential or both. Finally, evidence suggesting that intracellular Ca2+ levels might play a fundamental role in bacterial homeostasis is discussed, in particular the role of Ca2+ in the process of chemotaxis and in conferring bacteria heat stability. A vitamin K-dependent carboxylation reaction has been found in Escherichia coli which is similar to that reported in mammalian systems which results in gamma carboxylation of glutamate residues. Although all of the proteins containing gamma-carboxyglutamate described so far are involved in Ca2+ metabolism, the role of these proteins in Escherichia coli is unknown and remains to be elucidated.

Adenosine Triphosphatases↗
PubMed1980

The entry process as the target for energy input in active transport of alpha-aminoisobutyric acid by Mycobacterium phlei.

Mycobacterium phlei was shown to accumulate alpha-aminoisobutyric acid, establishing a concentration gradient of approximately 15,000-fold. The apparent affinity constant of the carrier for alpha-aminoisobutyric acid was 1.8 microM. The system exhibited a broad specificity provided two structural requirements were satisfied: the presence of a free amino and carboxyl group on the alpha carbon and the absence of a net charge. The role of energy coupling on the accumulation of alpha-aminoisobutyric acid was studied by two different kinds of experiments, the relative effects of the inhibitors on the rate of entry and the steady-state of accumulation, and a comparison of the efflux induced at the final steady state by the addition of (a) excess nonradioactive alpha-aminoisobutyric acid, (b) energy inhibitors, or (c) both. The results are consistent with the hypothesis that accumulation of alpha-aminoisobutyric acid is due to an increased rate of entry, the rate of exit not being affected by metabolic inhibitors.

Amino Acids↗
PubMed1980

Restoration of active transport of solutes and oxidative phosphorylation by naphthoquinones in irradiated membrane vesicles from Mycobacterium phlei.

Irradiation of the inverted membrane vesicles of Mycobacterium phlei with light at 360 nm inactivated the natural menaquinone [MK(9)(II-H)] and resulted in a loss of substrate oxidation, pH gradient, membrane potential, active transport of proline or calcium ions, and oxidative phosphorylation. Restoration of the protonmotive force and active transport occurred on addition of naphthoquinones such as vitamin K(1), menadione, or lapachol to the irradiated membrane vesicles. However, coupled phosphorylation was restored only by vitamin K(1). Menadione and lapachol did not act as uncoupling agents. The magnitude of the pH gradient and membrane potential in the quinone-restored system was a reflection of the rate of oxidation and was correlated with the rate of uptake of proline or Ca(2+). These results are consistent with the chemosmotic hypothesis proposed for the energy transducing mechanism for active transport and further demonstrate that the complete respiratory chain is not required to drive active transport. In contrast, the data suggest that in addition to the driving force (protonmotive force) necessary to establish oxidative phosphorylation, a specific spatial orientation of the respiratory components, such as the naphthaquinones, is essential for the utilization of the proton gradient or membrane potential or both. Bypass of electrons from the respiratory chain with menadione may explain the inability of this quinone to restore oxidative phosphorylation; however, lapachol restores oxidation by the same electron transport pathway as the natural menaquinone but fails to restore phosphorylation. Because all three quinones restore the protonmotive force, other factors that are discussed must be considered in understanding the mechanism of oxidative phosphorylation.

Biological Transport, Active↗
PubMed1979

Active transport of calcium in membrane vesicles from Mycobacterium phlei.

Active transport of calcium ions has been demonstrated in inside-out membrane vesicles from Mycobacterium phlei mediated by respiratory linked substrates as well as by ATP hydrolysis. The uptake of calcium exhibited an apparent Km of 80 microM and V of 16.6 nmol calcium uptake x min-1 x mg protein-1. A fortyfold concentration gradient for calcium ions was calculated for both the ATP-induced and the respiration-induced transport of calcium. Removal of coupling-factor-latent ATPase resulted in the complete loss of ATP-driven Ca2+ transport whereas the respiration-driven uptake was reduced by 40-50%. The uptake of calcium was inhibited by the proton conducting ionophores carbonylcyanide m-chlorophenylhydrazone and Gramicidin-D. The accumulated calcium was freely exchangeable with external calcium and was rapidly released by the addition of inhibitors of energy transduction, proton-translocating uncouplers or the ionophore A23187. The uptake of the weak base, methylamine, upon the oxidation of respiratory-linked substrates or the hydrolysis of ATP showed the generation of a protein gradient (inside acidic) which was partially collapsed on the addition of calcium ions. These results suggest that a Ca2+/H+ antiport mechanism may be responsible for the transport of calcium.

Adenosine Triphosphatases↗
PubMed1979

Resolution and reconstitution of active transport of calcium by a protein(s) from Mycobacterium phlei.

Membrane protein(s) responsible for the active transport of calcium in membrane vesicles from Mycobacterium phlei have been solubilized from membranes by sodium cholate treatment and partially purified using a hydrophobic resin. Reconstitution of calcium transport was demonstrated by reconstitution of detergent extracted membranes with the partially purified protein. The uptake of calcium in the reconstituted system was sensitive to proton-conducting uncouplers. Liposomes prepared with partially purified calcium translocating protein were capable of accumulating calcium. The uptake of calcium in this system occurred as a result of an artificial proton gradient generated by the reduction of entrapped ferricyanide with ascorbate-benzoquinone serving as a hydrogen carrier. The addition of the ionophore A23187 caused efflux of accumulated calcium in both native and proteoliposomal-reconstituted system.

Biological Transport, Active↗
PubMed1979

Tryptic proteolysis of coupling factor-latent ATPase from mycobacterium phlei. Theoretical modeling of structure-function relationships.

Trypsin treatment of solubilized coupling factor-latent ATPase from Mycobacterium phlei alters its subunit structure and functional properties. This coupling factor exhibits ATPase activity following trypsin treatment. Concurrently, both the ability of the enzyme to rebind to membranes depleted of coupling factor and its capacity for coupled phosphorylation are lost. The native alpha (64 000 dalton) subunit undergoes limited proteolytic digestion, and the delta (14 000 dalton) subunit is partially lost. During the course of tryptic proteolysis, the coupling factor molecule may exist in one of ten unique structural state (e.g. the native, ATPase-inactive molecule exists in the alpha alpha alpha state). Rigorous analysis of the experimental data by theoretical modeling provided information concerning the intermediate structural states leading to the fully ATPase-activated alpha" alpha" alpha" state under different conditions of trypsin treatment. The theoretical models of structure-function relationships that best-represented the experimental data predicted that the native coupling factor molecule contains three copies of the alpha (64 000 dalton) form of the alpha subunit, that the alpha" (58 000 dalton) alpha subunit species contributes maximally and the alpha' (61 000 dalton) form about half-maximally to ATPase activity, that membrane rebinding ability is proportional to the number of native alpha subunits in the enzyme, and that at least one native alpha subunit/molecule is required for full expression of coupled phosphorylation. These results indicate an essential role for the alpha subunit in the regulation of ATPase activity and in the ability of the solubilized coupling factor to rebind to depleted membranes.

Adenosine Triphosphatases↗
PubMed1979

Limited proteolysis of coupling factor-latent ATPase from Mycobacterium phlei. Effects of different enzymes and modifying agents.

The activation of the coupling factor-latent ATPase enzyme by tryptic proteolysis may resemble the activation of many proenzymes by limited proteolysis. The beta (53 000 dalton) subunit of solubilized coupling factor-latent ATPase from Mycobacterium phlei was selectively lost in some trypsin-treated samples. Since a concomitant loss of ATPase activity was not observed, the beta subunit may not be essential for ATPase catalytic activity. Treatment of solubilized coupling factor with chymotrypsin rapidly produced an A'-type (61 000 dalton) species from the native alpha (64 000 dalton) subunits with partial activation of the APTase enzyme. Secondary chymotryptic cleavage yielded an A"-type (58 000 dalton) species and a less-active enzyme. Storage of fresh coupling factor samples at -20degreeC in the presence of 4 mM MgCl2 with several freeze-thaw cycles resulted in loss of ATPase activity without apparent change in alpha subunit structure. Storage at 4 degrees C in the presence or absence of MgCl2 both decreased ATPase activity and generated A'-type alpha subunit species. Since presence was suspected. The peptide bonds first cleaved by trypsin, chymotrypsin, and the unknown protease are all apparantly located within the same small segment of alpha subunit polypeptide chain.

Adenosine Triphosphatases↗
PubMed1979

Isolation, purification, and reconstitution of a proline carrier protein from Mycobacterium phlei.

Membrane vesicles from Mycobacterium phlei contain carrier proteins for proline, glutamine, and glutamic acid. The transport of proline is Na+ dependent and required substrate oxidation. A proline carrier protein was solubilized from the membrane vesicles by treatment with cholate and Triton X-100. Electron microscopic observation of the detergent-treated membrane vesicles showed that they are closed structures. The detergent-extracted proteins were purified by means of sucrose density gradient centrifugation, followed by gel filtration and isoelectric focusing. A single protein with a molecular weight of 20,000 +/- 1000 was found on polyacrylamide gel electrophoresis. Reconstitution of proline transport was demonstrated when the purified protein was incubated with the detergent-extracted membrane vesicles. This reconstituted transport system was specific for proline and required substrate oxidation and Na+. The purified protein was also incorporated into liposomes, and proline uptake was demonstrated when energy was supplied as a membrane potential introduced by K+ diffusion via valinomycin. The uptake of proline was Na+ dependent and was inhibited by uncoupler or by sulfhydryl reagents.

Biological Transport↗