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Biomedical subjects

P J Butler

Publications and source records attributed to P J Butler.

At least 109 records · Page 6Linked to original sources
PubMed1990

Life-threatening complications during anaesthesia in a patient with a ventriculo-atrial shunt and pulmonary hypertension.

A 6-year-old patient with hydrocephalus who underwent revision of a ventriculo-atrial shunt is described. Anaesthesia was complicated by the occurrence of systemic hypertension and arterial hypoxaemia. The patient was subsequently found to have pulmonary hypertension secondary to recurrent pulmonary thromboembolism. The pathophysiological mechanisms for the patient's deterioration are discussed and the anaesthetic management of children with pulmonary hypertension is outlined. It is concluded that patients with a ventriculo-atrial shunt who present for surgery should be screened carefully for the presence of pulmonary hypertension.

Anesthesia, General↗
PubMed1989

Assembly of hybrid RNAs with tobacco mosaic virus coat protein. Evidence for incorporation of disks in 5'-elongation along the major RNA tail.

We have shown that during the reassembly of tobacco mosaic virus (TMV) RNA, with the coat protein supplied as a "disk preparation", the lengths of RNA protected from nuclease are "quantized" with steps which correspond to incorporation of the subunits from either a single or, more commonly, both rings of a disk. This interpretation has been challenged and it was suggested that the pattern was due to special, though unspecified features of the sequence of TMV RNA. To test whether the specific sequence of TMV RNA is important during the elongation, rather than just during nucleation, we have now followed growth of particles containing hybrid RNAs, with the TMV RNA origin of assembly but otherwise non-TMV sequences. We have prepared in vitro RNA transcripts containing heterologous RNA 5' to the origin of assembly sequence from TMV RNA, i.e. with a heterologous RNA tail in place of the natural major 5'-tail and no minor tail, and used these for assembly experiments. In each case we observe a banding pattern very similar to that which we had found with native TMV RNA and with a dominant quantum step of just over 100 bases, and sometimes also a step of 50 bases, strongly suggesting that this is not due to any feature of the TMV RNA. This same repeat is also visible even with a heterologous RNA chosen because it had a sequence repeat of 135 or 136 bases, confirming that the quantization is due to a feature of the elongation reaction and in no way to the RNA sequence being encapsidated. We have also followed elongation with the origin of assembly located 5' to the heterologous RNA. This leads to a slower elongation along this 3'-tail, after the initial rapid encapsidation of the origin RNA, which lacks any quantization of length protected. These results are fully compatible with the hypothesis we had advanced earlier, that the major growth along the 5'-tail is from performed aggregates ("disks") while the minor growth along the 3'-tail is from subunits in the "A-protein" adding singly or a few at a time.

Base Sequence↗
PubMed1988

The tobacco mosaic virus assembly origin RNA. Functional characteristics defined by directed mutagenesis.

The in vitro reassembly of tobacco mosaic virus (TMV) begins with the specific recognition by the viral coat protein disk aggregate of an internal TMV RNA sequence, known as the assembly origin (Oa). This RNA sequence contains a putative stem-loop structure (loop 1), believed to be the target for disk binding in assembly initiation, which has the characteristic sequence AAGAAGUCG exposed as a single strand at its apex. We show that a 75-base RNA sequence encompassing loop 1 is sufficient to direct the encapsidation by TMV coat protein disks of a heterologous RNA fragment. This RNA sequence and structure, which is sufficient to elicit TMV assembly in vitro, was explored by site-directed mutagenesis. Structure analysis of the RNA identified mutations that appear to effect assembly via a perturbation in RNA structure, rather than by a direct effect on coat protein binding. The binding of the loop 1 apex RNA sequence to coat protein disks was shown to be due primarily to its regularly repeated G residues. Sequences such as (UUG)3 and (GUG)3 are equally effective at initiating assembly, indicating that the other bases are less functionally constrained. However, substitution of the sequences (CCG)3, (CUG)3 or (UCG)3 reduced the assembly initiation rate, indicating that C residues are unfavourable for assembly. Two additional RNA sequences within the 75-base Oa sequence, both of the form (NNG)3, may play subsidiary roles in disk binding. RNA structure plays an important part in permitting selective protein-RNA recognition, since altering the RNA folding close to the apex of the loop 1 stem reduces the rate of disk binding, as does shortening the stem itself. Whereas the RNA sequence making up the hairpin does not in general affect the specificity of the protein-RNA interaction, it is required to present the apex signal sequence in a special conformation. Mechanisms for this are discussed.

Base Sequence↗
PubMed1988

Determination of the aggregate size in detergent solution of the light-harvesting chlorophyll a/b-protein complex from chloroplast membranes.

The molecular mass of an oligomeric integral membrane protein, the light-harvesting chlorophyll a/b-protein complex from the photosynthetic membranes of chloroplasts, has been determined in detergent solution by analytical ultracentrifugation and measurement of the density increment at constant chemical potential of all diffusible solutes. The technique used eliminates any problems resulting from detergent binding to the protein, is independent of the particular detergent used (in this case the nonionic n-octyl beta-D-glucopyranoside), and gives the apparent weight-average molecular mass at different protein concentrations, allowing extrapolation to zero concentration. It means that the solutions of the complex must be brought to dialysis equilibrium with the solvent detergent solution and also requires a reliable method for measuring the protein concentration, for which amino acid analysis was used. The detergent-solubilized complex was a trimer that dissociated into monomers and dimers at low protein concentration. The accurate concentration determinations also allowed the molar chlorophyll-to-protein ratio to be measured as 15, corresponding to 8 chlorophyll a and 7 chlorophyll b molecules.

Journal Article↗
PubMed1988

Chemoreceptor control of heart rate and behaviour during diving in the tufted duck (Aythya fuligula).

1. The role of chemoreceptors in the control of heart rate and behaviour during diving activity in the tufted duck was investigated in two ways. In a closed-loop experiment, ducks were exposed to ambient gas mixtures of varied composition during diving activity in an indoor tank. Characteristics of diving behaviour, heart rate and deep body temperature were monitored under hypoxic, hyperoxic and hypercapnic conditions and compared with those in air. Secondly, in an open-loop experiment the role of the carotid body (CB) chemoreceptors in the control of the responses to altered inspired gas composition and in the cardiac responses to extended and enclosed dives (Stephenson, Butler & Woakes, 1986) was investigated by chronic bilateral denervation of these receptors. 2. Heart rate during submersion was unaffected by inspired gas composition in control (data from intact and sham-operated ducks combined) and CB-denervated ducks, though diving behaviour was significantly modified in both groups of animals in response to altered inspired gas composition. Hypoxia and hypercapnia resulted in an increase in the proportion of total diving time spent breathing at the surface. The main effect of hypoxia (9-10% O2) was to reduce dive duration in control ducks and this effect was almost completely abolished after CB denervation. Hypercapnia (5-6% CO2) reduced dive duration less markedly than hypoxia but it greatly increased the duration of the inter-dive interval, effects which were not significantly influenced by CB denervation. Hyperoxia (40-45% O2) had very little effect on either behaviour or heart rate during diving, although deep body temperature was significantly elevated in this gas mixture during diving activity. There was also a less marked, but nevertheless significant, apparent hyperthermia during diving activity in air on an indoor tank but not on an outdoor pond. Conversely, there was a significant apparent hypothermia during diving activity under hypoxic conditions. 3. The CB chemoreceptors were shown to play a role in cardiac control during diving under certain circumstances. The duration of pre-dive tachycardia was significantly increased in hypoxia and this increase was abolished after CB denervation. The rate of development of bradycardia during extended and enclosed dives was slowed following CB denervation, though the initiation of the responses in extended and enclosed dives and the eventual attainment of sub-resting heart rates in enclosed dives were not prevented, indicating that other, as yet unidentified, sensory inputs are involved in cardiac control under these conditions.

Animals↗
PubMed1988

Effect of training on maximal oxygen uptake and aerobic capacity of locomotory muscles in tufted ducks, Aythya fuligula.

1. The effects of artificial swim training on maximal oxygen consumption and heart rate, as well as on the capillarity and oxidative capacity of locomotory muscles, have been studied in the tufted duck, Aythya fuligula. 2. The artificial training programme resulted in a 27% increase in maximal oxygen consumption, mainly as a result of an increase in muscle capillarity (20% increase in capillary/fibre ratio). In addition, activity of an oxidative enzyme, citrate synthase, increased (by 42%) and there was a significant transformation of fibre types in the lateral gastrocnemius muscle. 3. Altering the duration and nature of the training stimulus, for example flying and diving, can bring about different degrees of muscular adaptation, particularly in oxidative capacity.

Adaptation, Physiological↗
PubMed1988

The aerobic capacity of locomotory muscles in the tufted duck, Aythya fuligula.

The locomotory muscles of the tufted duck, Aythya fuligula (L.), were analysed for mass, aerobic and anaerobic enzyme activities, fibre-type proportions, capillarity, mitochondrial and myoglobin content. The estimated aerobic capacity of the muscles correlated well with the muscles' maximal oxygen uptake both when measured during swimming and when predicted for steady-state flight. The results suggest that exercise performance in birds cannot be predicted purely on the basis of muscle mass (see Butler & Woakes, 1985); the specific enzyme complement of each muscle must also be taken into account. The delivery of oxygen to mitochondria is facilitated by the dense capillarity and high myoglobin content of the muscles.

3-Hydroxyacyl CoA Dehydrogenases↗
PubMed1988

Regional distribution of blood flow during swimming in the tufted duck (Aythya fuligula).

The distribution of blood flow to a number of organs and tissues of the tufted duck was determined (by the microsphere technique) before and while the birds were swimming at close to their maximum sustainable velocity (i.e. at 0.69 +/- 0.01 ms-1). During swimming, oxygen uptake was twice the pre-exercise value. Cardiac output increased by 70%, there was no significant change in arterial blood pressure and total systemic conductance increased by 44%. There were no significant changes in blood flow to the brain, liver, adrenal glands, spleen and respiratory muscles. Not surprisingly, there were increases in blood flow to the heart (30% increase) and to the muscles of the hindlimbs (to 3.1 times the pre-exercise value). Significant reductions in flow occurred to various parts of the gastrointestinal tract (although not to the gastrointestinal tract as a whole), to the pancreas and to the pectoralis muscles. In the case of the flight musculature as a whole, the reduction was to approximately 40% of the values in the ducks before exercise. Thus, despite the fact that cardiac output was some three times lower than it would have been during flight, there was a clear redistribution of blood away from some visceral organs and inactive muscles during surface swimming in the tufted duck. This lends support to the suggestion that blood is selectively directed to the legs, as well as to the brain and central nervous system (CNS) and away from the visceral organs and inactive muscles during voluntary diving in these birds.

Animals↗
PubMed1987

Serum amyloid P component is the major calcium-dependent specific DNA binding protein of the serum.

Serum amyloid P component (SAP), a member of the highly conserved pentraxin family of plasma proteins, was found to be the only protein in whole normal or acute phase serum which underwent specific calcium-dependent binding to either single or double-stranded DNA immobilised on gel. Isolated purified SAP also bound to long chromatin, to H1-stripped chromatin and to native DNA in solution at physiological ionic strength. Pure SAP which had been immobilised on gel, specifically bound nucleosome core particles from solution. These observations strongly suggest that SAP may bind to extracellular chromatin and DNA in vivo and that this may be its physiological role.

Animals↗
PubMed1987

Heart rate in humans during underwater swimming with and without breath-hold.

Heart rate was monitored, by way of radiotelemetry, from 6 male subjects of mean age (+/- SE) 24 +/- 1 years and of mean mass 73.5 +/- 2.5 kg. Measurements were made in a 25 m pool at a water temperature of 28 degrees C. Resting heart rate was 67 +/- 3.7 beats X min-1 and when the subjects submerged themselves completely in the pool, but remained inactive, there was a prompt, gradual reduction in heart rate which reached 48 +/- 2.6 beats X min-1 within 30 sec and 40 +/- 2.6 beats X min-1 within 59 +/- 5.6 sec (maximum duration). When they propelled themselves under water for 33 sec by kicking their legs and breathed through a snorkel tube, heart rate increased progressively to a value of 118 +/- 4.1 beats X min-1 at 28 sec. However, when they performed the same manoeuvre while holding their breath, there was an initial increase in heart rate to 106 +/- 5.7 beats X min-1 within the first 10 sec. This was followed by a decline in heart rate which was more rapid than that recorded during inactive submersion and which eventually reached 48 +/- 4.4 beats X min-1 at mean underwater duration of 33 +/- 1.8 sec. It is concluded that during the first 10-15 sec of underwater breath-hold swimming in humans, the cardiovascular response (as indicated by heart rate) is similar to that seen during a similar level of exercise while breathing air. From then on there is a progressively more intense bradycardia which is probably indicative of an oxygen conserving response consisting of reduced perfusion of most of the body except the heart, CNS and active locomotory muscles. The degree and rate of onset of this proposed oxygen conserving response are influenced by the intensity of the exercise performed while under water and whether or not the period of underwater breath-hold swimming is preceded by exercise.

Adaptation, Physiological↗
PubMed1986

Essential features of the assembly origin of tobacco mosaic virus RNA as studied by directed mutagenesis.

The assembly origin of tobacco mosaic virus RNA contains three stable hairpin loops. Coat protein disks bind first to loop 1 (the 3' most) during virus assembly, but the whole region is coated in a concerted fashion even in conditions of limiting protein. It is shown by in vitro packaging assays using mutant assembly origin transcripts that rapid and specific assembly initiation occurs in the absence of loops 2 and 3, but is abolished on removal of loop 1. Deletion or alteration of the unpaired AAGAAGUCG sequence at the apex of loop 1 also abolishes rapid packaging; this sequence is therefore instrumental in disk binding. Alteration of this sequence to (A)9 leads to packaging at a very low rate (half time 12 hours) which is apparently non-sequence specific. Substitution of (CCG)3 evokes packaging with a half time of 3 hours, as compared to 15 seconds for the wild type assembly origin. These results suggest that the three-base G periodicity within this sequence element is an important feature in assembly nucleation.

Base Sequence↗
PubMed1986

Oligonucleotide binding to the coat protein disk of tobacco mosaic virus. Possible steps in the mechanism of assembly.

Binding of the oligoribonucleotides AAG, AAGAAG and AAGAAGUUG to the disk aggregate of tobacco mosaic virus coat protein has been studied in solution under conditions favourable for virus assembly. The two longer oligomers bind strongly with Kd around 1 microM, approach complete saturation of binding sites and cause the formation of long, nicked helical rods resembling the virus. It is suggested that the binding of these oligomers, with sequences chosen from the assembly origin of the viral RNA, simulates the tobacco mosaic virus assembly process. No binding could be detected for AAG, indicating that chain length is a crucial determinant in the interaction. The binding of AAGAAG to coat protein crystals is very much weaker than that observed in solution, and the crystals crack at high oligomer concentrations. The corresponding oligodeoxyribonucleotide, d(AAGAAG), shows no binding to the protein in solution; the interaction is extremely specific for RNA.

Base Sequence↗
PubMed1986

Studies of tobacco mosaic virus reassembly with an RNA tail blocked by a hybridised and cross-linked probe.

Segments of cloned cDNA to tobacco mosaic virus RNA, 150--300-bases long, have been hybridised and cross-linked to the RNA, which has then been used for reassembly experiments. This enables the elongation reaction, which does not encapsidate the double-stranded region generated, to be stopped at specific regions along the RNA and the resulting particles to be characterised, by measuring the lengths of the rods in the electron microscope. With hybridisation to the 3'-tail the entire RNA contiguous to the nucleation region is encapsidated, from the 5'-terminus up to the modified region. When the double-stranded region is on the 5'-side of the nucleation region, the mean length of the particles corresponds to a situation in which the double-stranded region is unable to enter the central hole of the growing rod, but the 3'-tail of the RNA is completely encapsidated. The longest particles hybridised on the 5'-tail (i.e. in a class longer than the mean length) show an effect complementary to those with a 3'-block, and have lengths which correspond to encapsidation from the modified region to the 3'-terminus, despite the continued presence of the 5'-tail up the rod. In all cases where there is a remaining 5'-tail the lengths observed can only be explained if elongation has occurred substantially, or probably completely, along the 3'-tail. Hence elongation must have occurred simultaneously along both the 5' and 3'-tails of the tobacco mosaic virus RNA after initiation on the internal nucleation region.

Binding Sites↗
PubMed1986

Salt-induced folding of sea urchin sperm chromatin.

We have studied the folding in solution of chromatin from sea urchin sperm, which has the longest reported nucleosome repeat length (approximately equal to 240 base pairs) and an H2B with a basic N-terminal extension. The same two size- and ionic-strength-dependent transitions in sedimentation behaviour are observed as for other chromatins (rat liver, chicken erythrocyte) with shorter DNA repeat lengths. The results support the notion of an essentially similar folding of chromatins with different linker lengths of 0-80 base pairs. We interpret the hydrodynamic data in terms of a solenoid with about six nucleosomes per turn, which has greater stability in sea urchin sperm than in the other cases. The higher values of the sedimentation coefficients for sea urchin sperm oligomers indicate lower frictional coefficients, which probably result from accommodating the additional linker DNA and the extra mass of larger H1 and H2B in the central hole and/or the surface interstices of the solenoid.

Animals↗
PubMed1986

Plasma catecholamines in the lesser spotted dogfish and rainbow trout at rest and during different levels of exercise.

The hypothesis that there is an increase in plasma catecholamines during exercise in fish and that they play an important role in the cardiovascular adjustments during exercise was investigated in the lesser spotted dogfish and rainbow trout. In resting fish plasma catecholamines were at a concentration of 10(-9)-10(-8) mol l-1. During spontaneous swimming in the dogfish, adrenaline increased by 3.3 times to 1.9 X 10(-8) mol l-1 and noradrenaline increased by 2.3 times to 3.2 X 10(-8) mol l-1. In rainbow trout swimming at a steady 1 body length s-1 (Ls-1) in a water channel, the levels of both amines decreased to approximately 25% of the resting values. When swimming to apparent exhaustion at approximately 2 Ls-1, adrenaline was 10 times the resting value at 1.4 X 10(-8) mol l-1, whereas noradrenaline was 2.2 times the resting value at 2.3 X 10(-8) mol l-1. Only after repeated burst swimming for 2-3 min did the levels of plasma catecholamines increase substantially above the resting values. In the dogfish, both amines were at 10(-7) mol l-1, whereas in the rainbow trout, noradrenaline was at 8.5 X 10(-8) mol l-1 and adrenaline was at 2 X 10(-7) mol l-1. These levels were compared with the concentrations of catecholamines used by other workers to elicit changes in the branchial vasculature, gas exchange at the gills or gas transport to the tissues. In lesser spotted dogfish, the levels of adrenaline and noradrenaline present in the plasma during spontaneous swimming have 80% and 50% of maximum effect on gill blood vessels, respectively, whereas in rainbow trout the levels present when swimming to apparent exhaustion have approximately 20% of maximum effect on the branchial vasculature. The levels present in the trout after repeated burst swimming have 40% of maximum effect on blood vessels in the gills. The difference between the dogfish and the trout may be related to the lack of innervation of the gill blood vessels in the former. Enhancement of gas exchange across the gills of rainbow trout can be demonstrated by using adrenaline at the concentration found after repeated burst swimming. It is possible, however, that the concentration of adrenaline found in the plasma of trout after swimming to apparent exhaustion may cause an increase in the concentration of oxygen in arterial blood, thus enhancing oxygen delivery to the tissues.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1986

Diving behaviour and heart rate in tufted ducks (Aythya fuligula).

Diving behaviour and heart rate were monitored in tufted ducks diving under circumstances which simulated various environmental conditions such as feeding under ice in winter. When distance to food was increased on a covered outdoor pond, dive duration increased proportionately, but it was calculated that time available for feeding was reduced during the longer-distance 'extended' dives. There was a gradual reduction in heart rate to 77.3 +/- 13.8 beats min-1, which is significantly lower than the resting value of 121.1 +/- 14.1 beats min-1, during the course of extended dives, suggesting that the ducks could gradually switch over to a 'classical' oxygen-conserving response during these prolonged voluntary dives. The duration of the pre-dive preparatory period was positively correlated with dive distance. When the ducks were briefly unable to resurface during an otherwise normal feeding dive in an indoor tank, a situation which may occur if they become disoriented under ice, there was an immediate switch to a full bradycardia. Reduction in heart rate during these 'enclosed' dives occurred only when the ducks were apparently aware of the situation and the rate of onset of bradycardia was very similar to that previously observed during involuntary submersion of tufted ducks. Minimum heart rate was the same at 46 beats min-1 after 15 s of enclosed dives and after 30 s of involuntary submersions, despite the differences in levels of activity in the two situations.

Animals↗