Biomedical subjects

P J Butler

Publications and source records attributed to P J Butler.

At least 73 records · Page 4Linked to original sources

PubMed1997

Characterisation of the major intrinsic protein (MIP) from bovine lens fibre membranes by electron microscopy and hydrodynamics.

The major intrinsic protein (MIP) from bovine lens fibre membranes has been purified from unstripped membranes using a single ion-exchange chromatography step (MonoS) in the non-ionic detergent octyl-beta-D-glucopyranoside (OG). SDS-PAGE has confirmed the purity of the preparation and thin-layer chromatographic analysis has shown that the protein is virtually lipid-free. To establish a stable and monodisperse protein sample, we exchanged OG with decyl-beta-D-maltopyranoside (DeM), another non-ionic detergent, by gel-filtration column chromatography. We conclude that the resulting protein/detergent complex is composed of four copies of MIP (a tetramer) and a detergent micelle. This conclusion is based on: (1) measurement of the weight-average molecular mass (Mw,app) of the protein moiety in the protein/detergent complex by sedimentation equilibrium; (2) measurement of the apparent molecular mass of the complexes formed by MIP in OG, in DeM, in dodecyl-beta-D-maltopyranoside (DoM) and in sodium dodecylsulphate (SDS) by gel filtration; (3) measurement of the apparent molecular mass of pure detergent micelles; (4) measurement of the predicted change in the molecular mass of the MIP/DeM complex after partial enzymatic proteolysis; and (5) measurement of the size and shape of the MIP/detergent complex by electron microscopy and single-particle analysis. Therefore, the tetragonal arrangement of MIP observed in both plasma membranes and junctional membranes in lens fibre cells is maintained in solution with non-ionic detergents.

Amino Acid Sequence↗

PubMed1997

Freeman-Sheldon (whistling face) syndrome. Anaesthetic and airway management.

We describe the anaesthetic management of Freeman-Sheldon syndrome in a two-and-a-half year old undergoing club foot correction. Following an inhalational induction using halothane, tracheal intubation by direct laryngoscopy proved impossible. A laryngeal mask airway was inserted and intubation with a 4.5 tracheal tube was successfully achieved using a fibreoptic bronchoscope passed through the LMA. The child had an uneventful anaesthetic course. A caudal epidural was used for postoperative analgesia and the child was discharged home on day two. The anaesthetic and airway management options of this syndrome are outlined.

Analgesia, Epidural↗

PubMed1997

The laryngeal mask airway: a comparison between two insertion techniques.

UNLABELLED: The purpose of the study was to compare the ease of insertion of the laryngeal mask airway using the standard uninflated approach or with a fully inflated cuff. Two hundred consecutive patients undergoing anesthesia using a laryngeal mask airway were randomized to have the laryngeal mask inserted using either method. Successful insertion was judged primarily by the clinical function of the airway. The number of insertion attempts to achieve a satisfactory airway and whether an alternative technique was required for success were recorded. On removal of the laryngeal mask, a blind observer noted the presence or absence of blood. Just before leaving the recovery room, each patient was asked whether they had a sore throat. Insertion technique made no difference with regard to first attempt success. However, the presence of blood on the removed masks (P < 0.01) and sore throat (P < 0.01) were less frequent in the inflated cuff group. We conclude that the inflated cuff insertion technique is an acceptable alternative to the standard approach and has the advantage of reducing the incidence of minor pharyngeal mucosal trauma, as evidenced by mucosal bleeding and sore throat. IMPLICATIONS: Insertion of the laryngeal mask airway with the cuff fully inflated is equally successful to the standard uninflated approach in experienced hands. The inflated technique was associated with less minor pharyngeal mucosal trauma and, consequently, a lower incidence of postoperative sore throat. This implies that the inflated technique would be acceptable to the general population of laryngeal mask users.

Anesthesia↗

PubMed1997

Physiology of diving of birds and mammals.

This review concentrates on the physiological responses, and their control, in freely diving birds and mammals that enable them to remain submerged and sometimes quite active for extended periods of time. Recent developments in technology have provided much detailed information on the behavior of these fascinating animals. Unfortunately, the advances in technology have been insufficient to enable physiologists to obtain anything like the same level of detail on the metabolic rate and physiological adjustments that occur during natural diving. This has led to much speculation and calculations based on many assumptions concerning usable oxygen stores and metabolic rate during diving, in an attempt to explain the observed behavior. Despite their shortcomings, these calculations have provided useful insights into the degree of adaptations of various species of aquatic birds and mammals. Many of them, e.g., ducks, smaller penguins, fur seals, and Weddell seals, seem able to metabolize aerobically, when diving, at approximately the same (if not greater) rate as they do at the surface. Their enhanced oxygen stores are able to support aerobic metabolism, at what would not be considered unusually low levels, for the duration of the dives, although there are probably circulatory readjustments to ensure that the oxygen stores are managed judiciously. For other species, such as the larger penguins, South Georgian shag, and female elephant seals, there is a general consensus that they must either be reducing their aerobic metabolic rate when diving, possibly by way of regional hypothermia, and/or producing ATP, at least partly, by anaerobiosis and metabolizing the lactic acid when at the surface (although this is hardly likely in the case of the female elephant seals). Circulation is the proximate regulator of metabolism during aerobic diving, and heart rate is the best single indicator of circulatory adjustment. During voluntary dives, heart rates range from extreme bradycardia to well above resting, reflecting metabolic performance. Efferent cardiac control is largely parasympathetic. Reflex cardiorespiratory responses are modulated by conditioning and habituation, but reflexes predominate during extended dives and during recovery, when gas exchange is maximized.

Adenosine Triphosphate↗

PubMed1996

The motile major sperm protein (MSP) from Ascaris suum is a symmetric dimer in solution.

The major sperm protein (MSP) of Ascaris suum mediates amoeboid motility by forming an extensive intermeshed system of cytoskeletal filaments analogous to that formed by actin in many amoeboid cells. We have used a combination of biochemical and NMR methods to show that, in contrast to actin, MSP exist in solution as a symmetrical dimer. This result has important implications for the mechanism of both MSP filament assembly and the recognition of different MSP isoforms in vivo.

Actin Cytoskeleton↗

PubMed1996

Flexible regions of RNA structure facilitate co-operative Rev assembly on the Rev-response element.

The oligomerisation of Rev on the Rev-response element (RRE) was studied using a series of model substrates. Only a monomer of Rev is able to bind efficiently to a high affinity site that is flanked by perfect duplex RNA. Addition of a bulge or a second stem structure adjacent to the high affinity site permits the co-operative incorporation of a second Rev molecule to the RNA. Model RREs carrying bulges can bind Rev with a higher degree of co-operativity than the native structure. Oligomerisation was efficient when the bulge was moved to the opposite strand of the duplex, but was severely impaired when the distance between the bulge and the high affinity site was increased by more than 8 bp. Rev can oligomerise at either end of the RNA-protein complex formed at the high affinity site; when the duplex flanking a high affinity site is disrupted by a bulge or a stem, oligomerisation proceeds in the direction of the disruption regardless of the orientation of the high affinity site. The results are consistent with the "molecular rheostat" model for RRE function, which suggests that Rev binding to the RRE is highly distributive and provides a sensitive measurement of intracellular Rev concentrations.

Base Sequence↗

PubMed1996

Equilibrium dissociation and unfolding of the dimeric human papillomavirus strain-16 E2 DNA-binding domain.

The equilibrium unfolding reaction of the C-terminal 80-amino-acid dimeric DNA-binding domain of human papillomavirus (HPV) strain 16 E2 protein has been investigated using fluorescence, far-UV CD, and equilibrium sedimentation. The stability of the HPV-16 E2 DNA-binding domain is concentration-dependent, and the unfolding reaction is well described as a two-state transition from folded dimer to unfolded monomer. The conformational stability of the protein, delta GH2O, was found to be 9.8 kcal/mol at pH 5.6, with the corresponding equilibrium unfolding/dissociation constant, Ku, being 6.5 x 10(-8) M. Equilibrium sedimentation experiments give a Kd of 3.0 x 10(-8) M, showing an excellent agreement between the two different techniques. Denaturation by temperature followed by the change in ellipticity also shows a concomitant disappearance of secondary and tertiary structures. The Ku changes dramatically at physiologically relevant pH's: with a change in pH from 6.1 to 7.0, it goes from 5.5 x 10(-8) M to 4.4 x 10(10) M. Our results suggest that, at the very low concentration of protein where DNA binding is normally measured (e.g., 10(-11) M), the protein is predominantly monomeric and unfolded. They also stress the importance of the coupling between folding and DNA binding.

Base Sequence↗

PubMed1996

Preoxygenation in children using expired oxygraphy.

We performed preoxygenation on 25 patients, aged 1-12 yr. End-tidal oxygen sampling was used to find the duration of preoxygenation required to reach an end-tidal oxygen fraction of 0.9. All children reached this end-point within 80 s, which was markedly more rapid than that observed in adult subjects. The clinical applications of this form of monitoring in children are discussed.

Anesthesia, General↗

PubMed1996

Postoperative analgesia in dental day case surgery. A comparison between Feldene "Melt' (piroxicam) and diclofenac suppositories.

A randomised, double-blind placebo-controlled trial was conducted to compare the analgesic efficacy of piroxicam as Feldene "Melt' with diclofenac suppositories following day case wisdom tooth extraction. Piroxicam was found to be equivalent to diclofenac in speed of onset and efficacy when given 1 h prior to surgery with no significant adverse effects. It is therefore possible to confer the benefits of postoperative analgesia from a non-steroidal anti-inflammatory drug whilst avoiding the inconvenience of the rectal route of administration.

Administration, Oral↗

PubMed1995

Incorporation of glutamine repeats makes protein oligomerize: implications for neurodegenerative diseases.

Many transcription factors and some other proteins contain glutamine repeats; their abnormal expansion has been linked to several dominantly inherited neuro-degenerative diseases. Having found that poly(L-glutamine) alone forms beta-strands held together by hydrogen bonds between their amide groups, we surmised that glutamine repeats may form polar zippers, an unusual motif for protein-protein interactions. To test this hypothesis, we have engineered a Gly-Gln10-Gly peptide into the inhibitory loop of truncated chymotrypsin inhibitor 2 (CI2), a small protein from barley seeds, by both insertion and replacement. Gel filtration resolved both mutant inhibitors into at least three fractions, which analytical ultracentrifugation identified as monomers, dimers, and trimers of the recombinant protein; the truncated wild-type CI2 formed only monomers. CD difference spectra of the dimers and trimers versus wild type indicated that their glutamine repeats formed beta-pleated sheets, while those of the monomers versus wild type were more suggestive of type I beta-turns. The CD spectra of all three fractions remained unchanged even after incubation at 70 degrees C; neither the dimers nor the trimers dissociated at this temperature. We argue that the stability of all three fractions is due to the multiplicity of hydrogen bonds between extended strands of glutamine repeats in the oligomers or within a beta-hairpin formed by the single glutamine repeat of each monomer. Pathological effects may arise when expanded glutamine repeats cause proteins to acquire excessively high affinities for each other or for other proteins with glutamine repeats.

Amino Acid Sequence↗

PubMed1995

Studies on nerves of the upper respiratory tract in the ferret and the mink.

The central localization of the superior laryngeal nerve (SLN), recurrent laryngeal nerve (RLN) and pharyngeal nerve (PHAR) in the ferret (Mustela putorius furo) and mink (Mustela vision) was determined by neuronal tract tracing techniques. The anterograde transport of horseradish peroxidase (HRP)/wheat germ aggluttinin conjugated HRP mixtures (WGA-HRP) revealed afferent fibres of the SLN projecting ipsilaterally in the tractus solitarius (TS) before terminating in the ipsilateral nucleus of the tractus solitarius (nTS). In both mustelids large concentrations of terminal reaction product were observed in the dorsal/dorsolateral and medial subnuclei of the nTS; however, at levels near obex significant projections of the SLN to the interstitial subnucleus were also observed. Caudal to obex, sparse terminal labelling was identified bilaterally in the nucleus commissuralis (n com). There were no labelled afferent projections of the SLN to the spinal trigeminal complex in either mustelid; neither was afferent terminal reaction product detected in the medulla oblongata following labelling of the RLN (in either the ferret or the mink) or PHAR (in the ferret). Projection of the SLN to areas of the nTS associated with upper airway functions, like swallowing and respiration, suggest a substantial role for this nerve in the initiation and control of airway reflexes. Extensive labelling of the nucleus ambiguus (nA) was revealed following HRP/WGA-HRP application to the PHAR, RLN and SLN and there was some evidence of a topographical arrangement of neuronal cell bodies in the nA, relative to the different nerves, in the ferret. In the case of the SLN, retrogradely labelled cells were also identified in the dorsal motor nucleus of the vagus (DmnX), nTS and reticular formation (rf). Electrical stimulation of the SLN produced periods of apnoea/ataxic breathing in both mustelids. As well as respiratory effects, stimulation of the SLN often caused bradycardia in the mink, but not in the ferret, in which heart rate generally remained unchanged. This difference did not reflect anatomical differences in the central localization of the SLN.

Afferent Pathways↗

PubMed1995

Implantable data logging system for heart rate and body temperature: its application to the estimation of field metabolic rates in Antarctic predators.

The metabolic rate of free-ranging animals is difficult to measure, but of great importance in understanding the interactions of a species with its environment. Heart rate can, if correctly validated and calibrated, give an estimate of metabolic rate, with both a fine time resolution and over long periods. The telemetry of heart rate is well documented, but is not appropriate over long ranges (possibly several thousands of kilometres) or for diving species. An implantable data logger has therefore been developed for the long term recording of heart rate and body temperature. The logger is built using hybrid and ASIC construction techniques, weighs 20 g and measures 55 x 24 x 6 mm. The device is programmable, and its solid-state memory holds over 70 days of data if, for example, heart rate is counted and stored every minute. Current consumption is 155 microA while logging, 50 microA during a programmable initial delay period, and less than 1 microA when the logger closes down after filling its memory. These loggers have been deployed for two field seasons in gentoo penguins, black-browed albatross and fur seals.

Animals↗

PubMed1995

Development of metabolic enzyme activity in locomotor and cardiac muscles of the migratory barnacle goose.

Preflight development of the goslings was typified by rapid increases in the mitochondrial enzymes of the semimembranosus and heart ventricular muscles resulting in near-adult values by 3 wk of age. In contrast, aerobic capacity of the pectoralis muscle initially developed slowly but showed a rapid increase between 5 and 7 wk of age, in preparation for becoming airborne. Activities of glycolytic enzymes in the pectoralis muscle showed similar patterns of development as those found for the aerobic enzymes, except for hexokinase, which was low at all ages, indicating an adaptation for catabolism of both intracellular glycogen and plasma fatty acids in preference to plasma glucose. Muscle mass specific activity of citrate synthase in the pectoralis increased by only 33% from goslings during the first few days of flight, compared with premigratory geese. Activities of anaerobic glycolytic enzymes in the ventricles were low, but values for hexokinase, which is involved in the phosphorylation of plasma glucose, developed rapidly. Values for lactate dehydrogenase were also high, reflecting the capacity of the heart to catabolize plasma lactate. Substrate flux supplied by carnitine palmitoyltransferase and oxoglutarate dehydrogenase (OGD), in the pectoralis muscles of the premigratory geese, appears to have the smallest excess capacities to meet the requirements of sustained aerobic flight. The average maximum oxygen uptake for premigratory geese during flight, as indicated by values for OGD, is calculated to be 484 ml O2/min (or 208 ml O2.min-1.kg-1).

Aging↗

PubMed1994

A molecular rheostat. Co-operative rev binding to stem I of the rev-response element modulates human immunodeficiency virus type-1 late gene expression.

The complete biologically active human immunodeficiency virus type-1 (HIV-1) rev-response element (RRE) RNA is 351 nucleotides (nt) in length, and includes an extra 58 nt on the 5' end and 59 nt on the 3' end beyond the sites proposed in the original models for the RRE secondary structure. The extra sequences are able to form a duplex structure which extends Stem I. The presence of an elongated Stem I structure in the RRE RNA was confirmed by nuclease mapping experiments. Nuclease protection experiments have shown that rev binds to restricted regions of the RRE, including the high affinity site located at the base of Stem IIb and along the length of the Stem I region. The three large stem-loop structures which protrude from Stem I and Stem IIb (Stems IIc, III+IV and V) remain accessible to nucleases even in the presence of a large excess of protein. Gel-retardation experiments show that the truncations of Stem I reduced the total number of rev molecules that can bind co-operatively and with high affinity to the RRE RNA. To test whether the elongated Stem I structure is required for maximal rev activity, a series of truncations which progressively reduced the length of Stem I was introduced into an HIV-1 derived reporter plasmid. In the presence of rev and a functional RRE, there is an increase in the levels of gag and env mRNA in the cytoplasm and a decrease in levels of tat and rev mRNAs. Each of the truncations in Stem I reduced the rev responses, with the longest truncations producing the greatest losses of activity. The data suggest that the RRE acts as a "molecular rheostat" designed to detect rev levels during the early stages of the HIV growth cycle.

Base Sequence↗

PubMed1994

Temperature and pH effects on the oxygen equilibrium curve of the thoroughbred horse.

A new oxygen equilibrium curve is defined for the Thoroughbred horse under standard conditions of 37 degrees C, pH = 7.4 and PCO2 = 5.33 kPa. The "standard" P50 for the Thoroughbred is, at 2.83 +/- 0.04 (SE of mean) kPa, significantly lower than that found for the Hanoverian horse (3.17 +/- 0.03 kPa) by Clerbaux et al. (Can. J. Vet. Res. 50: 188-192, 1986), and lower than other values for horses in the literature. Using data from Butler et al. (J. Exp. Biol. 179: 159-180, 1993), curves were also constructed, in vitro, under simulated conditions of intense exercise to examine the individual effects of pH, temperature and PCO2 on the standard curve. The fixed acid Bohr coefficient is similar to that in humans (-0.41) whereas the temperature coefficient is, at 0.019, lower than that for humans. The coefficients were shown to be saturation dependent.

Animals↗

PubMed1994

Binding of pentraxins to different nuclear structures: C-reactive protein binds to small nuclear ribonucleoprotein particles, serum amyloid P component binds to chromatin and nucleoli.

Binding of the human pentraxin plasma proteins, C-reactive protein (CRP) and serum amyloid P component (SAP), to the nuclei of human cells was studied using whole acute phase serum as the source of the proteins and confocal immunofluorescence microscopy. CRP and SAP clearly bound to distinct, different structures. Double staining with MoAbs to the Sm D and Sm B/B' components of small nuclear ribonucleoproteins confirmed that CRP bound exclusively to these particles. As expected, SAP bound to chromatin and, in addition, binding to the nucleolus was observed for the first time. These interactions demonstrated under relatively physiological conditions, with native pentraxins unseparated from serum and with nuclear constituents in situ, are likely to be of functional importance in vivo.

Animals↗

PubMed1994

An algorithm to describe the oxygen equilibrium curve for the thoroughbred racehorse.

An algorithm to describe the oxygen equilibrium curve (OEC) of the Thoroughbred horse was derived from raw oxygen equilibrium curve data obtained under standard conditions of temperature, pH and PCO2 (Smale and Butler, 1994). This algorithm was derived by a curve-fitting procedure based on the algorithm for human blood produced by Kelman (1966). The temperature, fixed acid and net CO2 coefficients were then incorporated in the algorithm to enable the accurate calculation of % saturation from any combination of PO2, temperature, pH and PCO2. The algorithm was checked using blood gas data obtained from in vivo treadmill exercise tests as well as a standardised breathing test whereby horses inhaled several different gas mixtures. This algorithm proved more accurate for the Thoroughbred horse than that derived by Kelman.

Algorithms↗

PubMed1994

Gas exchange of captive freely diving grey seals (Halichoerus grypus).

When at sea, phocids dive for long periods and spend a high percentage of their time submerged. This behaviour requires some combination of an increased oxygen storage capacity, rapid oxygen loading at the surface and reduced oxygen utilisation when submerged. To assess these adaptations, breath-by-breath ventilation was studied in four adult grey seals (two male, two female, 160-250 kg), freely diving in a large outdoor tank where surface access was restricted to one breathing hole. The dive patterns obtained were similar to those recorded from freely diving wild grey seals. Respiratory frequency during the surface periods was 40% higher than that estimated from allometric relationships (19.4 +/- 0.7 breaths min-1), and tidal volume (6.3 +/- 1.21) was approximately five times higher than that estimated from allometric relationships. These adaptations produce a high minute volume and enable gas exchange to occur at the surface. Mean oxygen consumption rate (VO2, measured for a dive+surface cycle) decreased with increasing dive duration. The aerobic dive limit was estimated as 9.6 min for a 150 kg grey seal (using the overall average VO2 of 5.2 ml O2 min-1 kg-1), which is consistent with results from freely diving wild grey seals (only 6% of dives exceeded 10 min). End-tidal oxygen values varied during a surface period, following a U-shaped curve, which suggests that there is limited oxygen uptake from the lung and/or blood oxygen stores during dives. This result was unexpected and indicates that these seals are utilising substantial physiological responses to conserve oxygen, even during shallow voluntary diving.

Animals↗