Search PubMed⌕ Search

Biomedical subjects

D H Evans

Publications and source records attributed to D H Evans.

At least 37 records · Page 2Linked to original sources
PubMed2000

Estimating normal and pathological dynamic responses in cerebral blood flow velocity to step changes in end-tidal pCO2.

The regulation of cerebral blood flow (CBF) following changes in arterial blood pressure (ABP) and end-tidal pCO2 (EtCO2) are of clinical interest in assessing cerebrovascular reserve capacity. Linear finite-impulse-response modelling is applied to ABP, EtCO2 and CBF velocity (CBFV, from transcranial Doppler measurements), which allows the CBFV response to ideal step changes in EtCO2 to be estimated from clinical data showing more sluggish, and additional random variations. The confounding effects of ABP changes provoked by hypercapnia on the CBFV are also corrected for. Data from 56 patients suffering from stenosis of the carotid arteries (with normal or diminished cerebrovascular reactivity to EtCO2 changes--CVRCO2) were analysed. The results show the expected significant differences (p < 0.05) between EtCO2 steps up and down, the significant contribution from ABP variation, and also differences in the dynamic responses of patients with reduced CVRCO2 (p < 0.01 after 10 s). For the latter the CBFV response appears exhausted after about 15 s, whereas for normals CBFV continues to increase. While dispersion of individual step responses remains large, the method gives encouraging results for the non-invasive study of compromised haemodynamics in different patient groups.

Aged↗
PubMed2000

The effects of beam shape on the ability to predict changes in vessel size from Doppler signal power.

Theory suggests that, under certain ideal circumstances, the power of a Doppler signal is proportional to the size of the vessel from which it is recorded and can, therefore, be used to assess the scale of any changes in vessel size that occur during clinical recordings of cerebral blood flow. However, the relationship between signal power and vessel size depends on the intensity of the incident beam being uniform across the vessel. This is unlikely to be the case for cerebral vessels insonated by commercial transducers. A model was used to estimate the signal power received from vessels insonated by a beam passing through a homogeneous medium, and also by beams passing through each of five samples of temporal bone. In each case, the effects of initial vessel size and various changes in cross-sectional area were investigated for different vessel positions in the beam. The results for the beam paths through bone predict that the power change arising from a change in vessel cross-sectional area is between 5% and 75% smaller than that occurring in a uniform beam. If these results are representative of those arising for an in vivo change in middle cerebral artery size, then the potential magnitude of the error illustrates the caution that may need to be applied when interpreting changes in Doppler signal power.

Adult↗
PubMed2000

The effects of temporal bone on transcranial Doppler ultrasound beam shape.

A knowledge of beam shape is desirable for many Doppler ultrasound applications, and is especially important for transcranial Doppler recordings where the beam may undergo significant distortion by the skull. Although it may not be possible to determine the precise beam shape for individual cases due to variations in the physical characteristics of the media in the beam path, information about the range of beam shapes that are likely to arise for in vivo recordings from the middle cerebral artery may in future allow limits of uncertainty to be derived, and may even allow partial correction in some cases. In order to assess the potential intersubject variation in beam sensitivity across the middle cerebral artery, the beam shapes generated by four commercial Transcranial Doppler transducers and the effects on beam shape of four different samples of temporal bone were investigated. The results demonstrate that there seems to be relatively little difference in the beam shapes of commercial transducers, but that the distortion effects of temporal bone are variable and unpredictable.

Adult↗
PubMed2000

Using doppler signal power to detect changes in vessel size: a feasibility study using a wall-less flow phantom.

The power of a Doppler signal is theoretically proportional to the volume of blood within the sample volume of an ultrasound (US) beam and, hence, may provide a means of detecting in vivo changes in the cross-sectional area of cerebral vessels, such as the middle cerebral artery. The purpose of this study was to examine the relationship between power and vessel size for signals recorded from a wall-less flow phantom. The results demonstrate the importance for the in vitro case of maximising the received signal power for each channel to obtain the true relationship between power and size, and show that a nonproportional relationship observed between the two parameters is primarily caused by high-pass filtering and nonuniform insonation. In addition, an investigation of the reproducibility of power values after transducer repositioning has shown that variation occurs even when extreme care is taken to maximise the received signal intensity. The implications of these results for the in vivo use of the Doppler signal power method are discussed.

Blood Flow Velocity↗
PubMed2000

Grading of cerebral autoregulation in preterm and term neonates.

The response of cerebral blood flow velocity to a single spontaneous transient rise in blood pressure was studied to grade the cerebral autoregulatory response of newborns. Blood pressure was measured continuously through an umbilical or peripheral arterial catheter; continuous flow velocity recordings were taken from the middle cerebral artery using continuous wave Doppler ultrasound. From a cohort of 62 healthy term and preterm neonates, 325 transients in mean arterial blood pressure and mean cerebral blood flow velocity were identified for analysis using a foot-seeking algorithm. An initial classification of active or impaired autoregulation was given to each transient using a self-clustering technique. The grading of the transients was studied by examining the slope of the return of the cerebral blood flow velocity to baseline. Negative slopes indicate a normal autoregulation; slopes of 0 or greater indicate an absence of autoregulation. This classification was in agreement with the self-clustering method (Cohen's kappa = 0.94, P<0.0001). The relationship between the autoregulatory response assessed by the grading method and gestational age, postnatal age, and PCO(2) was examined using linear regression analysis. A significant relationship with gestational age (P = 0.002) but not PCO(2) (P = 0.06) or postnatal age (P = 0.14) was evident.

Blood Flow Velocity↗
PubMed2000

A comparison of colour duplex ultrasonography, papaverine testing and common femoral Doppler waveform analysis for assessment of the aortoiliac arteries.

OBJECTIVE: to study the "accuracy" of aortoiliac colour duplex ultrasonography. DESIGN: prospective study. SETTING: vascular laboratory, University Hospital. METHODS: a total of 25 aortoiliac stenoses were studied in 23 patients. For each iliac segment, colour duplex ultrasound, papaverine testing, hyperaemic common femoral Doppler waveform analysis and hyperaemic testing using a thigh pressure cuff were performed. A velocity ratio of two was used to indicate a significant 50% diameter-reducing stenosis, but the velocity differences across stenoses as well as various characteristics of the hyyperaemic common femoral waveform were also studied. Retrospective receiver-operator characteristics and Kappa values were used for analysis. RESULTS: the Kappa agreement between ultrasonography and papaverine testing was 0.12 using peak systolic velocity ratios and 0.8 using hyperaemic peak systolic velocity differences. Hyperaemic common femoral pulsatility (PI) and resistance index (RI) both gained a Kappa level of 0.60. The reactive hyperaemia produced by a thigh cuff was more pronounced than that produced by papaverine. CONCLUSION: although the velocity ratio did not appear to perform well against the papaverine test, its apparent over-sensitivity calls into question the sensitivity of papaverine testing itself. The hyperaemic velocity difference at the stenosis or the hyperaemic PI or RI at common femoral level appear useful, non-invasive indicators of significant aortoiliac arterial disease.

Aged↗
PubMed2000

Multivariate dynamic analysis of cerebral blood flow regulation in humans.

The contributions of beat-to-beat changes in mean arterial blood pressure (MABP) and breath-by-breath fluctuations in end-tidal CO2 (EtCO2) as determinants of the spontaneous variability of cerebral blood flow velocity (CBFV) were studied in 16 normal subjects at rest. The two input variables (MABP and EtCO2) had significant cross-correlations with CBFV but not between them. Transfer functions were estimated as the multivariate least mean square finite impulse response causal filters. MABP showed a very significant effect in explaining CBFV variability (p < 10(-11), Fisher's aggregated-p test) and the model mean square error was significantly reduced (p < 0.001) by also including the contribution of EtCO2. The estimated mean CBFV step response to MABP displayed the characteristic return to baseline caused by the cerebral autoregulatory response. The corresponding response to EtCO2 showed a gradual rise taking approximately 10 s to reach a plateau of 2.5%/mmHg. This study demonstrated that spontaneous fluctuations in EtCO2 can help to explain the CBFV variability at rest if appropriate signal processing techniques are employed to address the limited power and bandwith of the breath-by-breath EtCO2 signal.

Adult↗
PubMed2000

Assessment of the thigh cuff technique for measurement of dynamic cerebral autoregulation.

BACKGROUND AND PURPOSE: Dynamic methods of measuring cerebral autoregulation have become an accepted alternative to static evaluation. This article aims to describe a set of data collected from healthy volunteers by a dynamic method, the purpose being to qualify and quantify expected results for those who may be designing a study using this technique. METHODS: Cerebral blood flow velocity (CBFV) (measured by transcranial Doppler) and arterial blood pressure (Finapres) were recorded in 16 normal subjects before, during, and after the induction of a blood pressure drop (release of bilateral thigh cuffs). This procedure was repeated 6 times for each subject. A mathematical model was applied to the data to generate an autoregulatory index (ARI) with values between 0 and 9. RESULTS: The ARI values for this sample population follow a normal distribution, with a mean+/-SD of 4.98+/-1.06 (n=15). Analysis of the cumulative mean ARI values of all subjects showed an exponential-type convergence of ARI toward the sample mean as the number of test iterations increased. The population average blood pressure drop on thigh cuff release was 26.4+/-7.1 mm Hg (n=16), occurring in 4.6+/-1. 7 seconds. The corresponding population average drop for CBFV was 15. 6+/-5.8 cm/s, taking 2.5+/-1.0 seconds. No significant trend was noted in the measurements as the number of test iterations increased. The correlation between the predicted and actual CBFV, having a mean value of 0.76+/-0.19, showed evidence of a nonlinear relationship to ARI values. Significant correlation was also found between ARI and (1) arterial blood pressure before cuff release and (2) the magnitude of the drop in CBFV on cuff release. CONCLUSIONS: The distribution of ARI values is not significantly different from normal. At least 3 iterations of the test procedure should be performed and averaged to obtain the mean ARI for each subject. There is no significant evidence of physiological accommodation as the number of test iterations increases. The effects of mean blood pressure and the magnitude of the change in CBFV should be considered as possible covariates when ARI data are analyzed.

Adult↗
PubMed2000

Dynamic cerebral autoregulation in sick newborn infants.

The sick newborn infant is vulnerable to brain injury and impaired cerebral autoregulation is thought to contribute to this. Coherent averaging is a method of measuring the dynamic cerebral autoregulatory response that is particularly suitable for neonates. We used this method in combination with a measure of the gradient of the cerebral blood flow velocity (CBFV) response following transient blood pressure (BP) peaks to study dynamic autoregulation in infants undergoing intensive care. Term and preterm infants at high risk of neurologic injury were compared with a control group of infants, also undergoing intensive care. Simultaneous video-EEG, CBFV (using transcranial Doppler), and arterial blood pressure measurements were obtained intermittently during a study period of at least 2 h. Cerebral autoregulatory response curves were constructed for high risk and control groups. Intact cerebral autoregulation produces a characteristic response consisting of a brief period when CBFV follows arterial blood pressure but quickly returns to baseline value. An impaired autoregulatory response shows CBFV mirroring the arterial blood pressure curve closely. Thirteen high-risk infants, who also had seizures (10 term and 3 preterm) and 12 control infants (6 term and 6 preterm) were studied. Autoregulation was absent in high-risk term and preterm infants. It was also absent in preterm control infants. Term, neurologically healthy infants undergoing intensive care have an intact autoregulatory response. The constant passive response seen in high-risk infants may reflect the severity of the underlying neurologic disease.

Blood Flow Velocity↗
PubMed2000

Effects of environmental salinity on Na(+)/K(+)-ATPase in the gills and rectal gland of a euryhaline elasmobranch (Dasyatis sabina).

Changes in Na(+)/K(+)-ATPase activity and abundance associated with environmental salinity were investigated in the gills and rectal gland of the Atlantic stingray Dasyatis sabina. Using a ouabain-specific ATPase assay and western blotting, we found that stingrays from fresh water had the highest activity and highest relative abundance of Na(+)/K(+)-ATPase in the gills. Using immunohistochemistry, we also found that gills from freshwater stingrays had the greatest number of Na(+)/K(+)-ATPase-rich cells. When freshwater stingrays were acclimated to sea water for 1 week, the activity and abundance of Na(+)/K(+)-ATPase and the number of Na(+)/K(+)-ATPase-rich cells decreased in the gills. In seawater stingrays, the branchial activity and abundance of Na(+)/K(+)-ATPase and the number of Na(+)/K(+)-ATPase-rich cells were further reduced. In rectal glands, the activity and abundance of Na(+)/K(+)-ATPase were lower in freshwater animals than in seawater-acclimated and seawater stingrays, both of which had equivalent levels. These findings suggest that salinity-associated changes in gill and rectal gland Na(+)/K(+)-ATPase activity are due to changes in the abundance of Na(+)/K(+)-ATPase. We conclude that the gills may be important for active ion uptake in fresh water, while the rectal gland is important for active NaCl excretion in sea water. The results from this study are the first to demonstrate an effect of environmental salinity on Na(+)/K(+)-ATPase expression in the gills and rectal gland of an elasmobranch.

Animals↗
PubMed2000

Activation of MeIQ (2-amino-3,4-dimethylimidazo- [4,5-f]quinoline) by sequence variants of recombinant human cytochrome P450 1A2.

Understanding the relationships between the sequences and catalytic activities of P450 enzymes that catalyze the bioactivation of mutagens and carcinogens is an important goal in mutation research. Escherichia coli strain DJ4309 expresses recombinant human P450 1A2 and activates promutagens such as MeIQ (2-amino-3, 4-dimethylimidazo[4,5-f]quinoline), as measured by induction of reverse mutations detected as lacZ(+) colonies on minimal lactose (ML) plates. Pools of P450 1A2 mutants were constructed by polymerase chain reaction (PCR) mutagenesis of putative substrate recognition sites (SRSs). Cultures of individual clones were patched onto MeIQ/ML plates and the growth of revertant microcolonies within each patch was inspected after 2 days of incubation. Beginning with a pool of several thousand clones, we identified 25 distinct P450 1A2 SRS variants with altered activities. In this study, the MeIQ dose-responses of all the variants are reported. The implications of the results are considered with reference to published models of the protein's structure.

Biotransformation↗
PubMed1999

The complete genome sequence of shope (rabbit) fibroma virus.

We have determined the complete DNA sequence of the Leporipoxvirus Shope fibroma virus (SFV). The SFV genome spans 159.8 kb and encodes 165 putative genes of which 13 are duplicated in the 12.4-kb terminal inverted repeats. Although most SFV genes have homologs encoded by other Chordopoxvirinae, the SFV genome lacks a key gene required for the production of extracellular enveloped virus. SFV also encodes only the smaller ribonucleotide reductase subunit and has a limited nucleotide biosynthetic capacity. SFV preserves the Chordopoxvirinae gene order from S012L near the left end of the chromosome through to S142R (homologs of vaccinia F2L and B1R, respectively). The unique right end of SFV appears to be genetically unstable because when the sequence is compared with that of myxoma virus, five myxoma homologs have been deleted (C. Cameron, S. Hota-Mitchell, L. Chen, J. Barrett, J.-X. Cao, C. Macaulay, D. Willer, D. Evans, and G. McFadden, 1999, Virology 264, 298-318). Most other differences between these two Leporipoxviruses are located in the telomeres. Leporipoxviruses encode several genes not found in other poxviruses including four small hydrophobic proteins of unknown function (S023R, S119L, S125R, and S132L), an alpha 2, 3-sialyltransferase (S143R), a protein belonging to the Ig-like protein superfamily (S141R), and a protein resembling the DNA-binding domain of proteins belonging to the HIN-200 protein family S013L). SFV also encodes a type II DNA photolyase (S127L). Melanoplus sanguinipes entomopoxvirus encodes a similar protein, but SFV is the first mammalian virus potentially capable of photoreactivating ultraviolet DNA damage.

Amino Acid Sequence↗
PubMed1999

Plasmid-mediated expression of the UmuDC mutagenesis proteins in an Escherichia coli strain engineered for human cytochrome P450 1A2-catalyzed activation of aromatic amines.

The mutagenic actions of many chemicals depend on the activities of bacterial "mutagenesis proteins", which allow replicative bypass of DNA lesions. Genes encoding these proteins occur on bacterial chromosomes and plasmids, often in the form of an operon (such as umuDC or mucAB) encoding two proteins. Many bacterial strains used in mutagenicity testing carry mutagenesis protein genes borne on plasmids, such as pKM101. Our objective was to introduce mutagenesis protein function into Escherichia coli strain DJ4309. This strain expresses recombinant human cytochrome P450 1A2 and NADPH-P450 reductase and carries out the metabolic conversion of aromatic and heterocyclic amines into DNA-reactive mutagens. We discovered that many mutagenesis-protein plasmids severely inhibit the response of strain DJ4309 to 2-amino-3,4-dimethylimid-azo[4,5-f]quinoline (MeIQ), a typical heterocyclic amine mutagen. Among many plasmids examined, one, pGY8294, a pSC101 derivative carrying the umuDC operon, did not inhibit MeIQ mutagenesis. Strain DJ4309 pGY8294 expresses active mutagenesis proteins, as shown by its response to mutagens such as 1-nitropyrene and 4-nitroquinoline 1-oxide (4-NQO), and is as sensitive as the parent strain DJ4309 to P450-dependent mutagens, such as MeIQ and 1-aminopyrene.

Amines↗
PubMed1999

Natriuretic peptide binding sites in the brain of the Atlantic hagfish, Myxine glutinosa.

We have previously used immunohistochemistry to show that the brain of the hagfish, Myxine glutinosa, contains a rich distribution of natriuretic peptide-immunoreactive elements with the densest distribution occurring in the telencephalon and the diencephalon. In this study, the distribution of (125)I-rat ANP and (125)I-porcine CNP binding sites was determined in the brain of M. glutinosa. The binding pattern of (125)I-rat ANP and (125)I-porcine CNP showed similarities; however, some differences were observed in the olfactory bulb and the caudal brain regions. Specific (125)I-rat ANP and (125)I-porcine CNP binding was observed in the olfactory bulb, outer layers of the pallium, and in regions of the diencephalon. Very little specific binding was observed in the habenula and the primordium hippocampi. In the diencephalon, a distinct zone of specific (125)I-rANP binding separated a region of moderate binding in the lateral regions of the diencephalon from the thalamic and hypothalamic nuclei. Moderate levels of specific (125)I-rANP binding were observed in the mesencephalon and medulla oblongata; little or no (125)I-porcine CNP binding was observed in these regions. The data, in combination with previous immunohistochemical studies, show that the natriuretic peptide system of the hagfish brain is well-developed and suggest that natriuretic peptides have a long evolutionary history as neurotransmitters and/or neuromodulators in the vertebrate brain. J. Exp. Zool. 284:407-413, 1999.

Animals↗
PubMed1999

DNA binding and aggregation properties of the vaccinia virus I3L gene product.

The vaccinia virus I3L gene encodes a single-stranded DNA-binding protein which may play a role in viral replication and genetic recombination. We have purified native and recombinant forms of gpI3L and characterized both the DNA-binding reaction and the structural properties of DNA-protein complexes. The purified proteins displayed anomalous electrophoretic properties in the presence of sodium dodecyl sulfate, behaving as if they were 4-kDa larger than the true mass. Agarose gel shift analysis was used to monitor the formation of complexes composed of single-stranded DNA plus gpI3L protein. These experiments detected two different DNA binding modes whose formation was dependent upon the protein density. The transition between the two binding modes occurred at a nucleotide to protein ratio of about 31 nucleotides per gpI3L monomer. S1 nuclease protection assay revealed that at saturating protein densities, each gpI3L monomer occludes 9.5 +/- 2.5 nucleotides. In the presence of magnesium, gpI3L promoted the formation of large DNA aggregates from which double-stranded DNA was excluded. Electron microscopy showed that, in the absence of magnesium and at low protein densities, gpI3L forms beaded structures on DNA. At high protein density the complexes display a smoother and less compacted morphology. In the presence of magnesium the complexes contained long fibrous and tangled arrays. These results suggest that gpI3L can form octameric complexes on DNA much like those formed by Escherichia coli single-stranded DNA protein. Moreover, the capacity to aggregate DNA may provide an environment in which hybrid DNA formation could occur during DNA replication.

Cloning, Molecular↗
PubMed1999

Vaccinia virus DNA polymerase promotes DNA pairing and strand-transfer reactions.

Vaccinia virus infection results in the synthesis of a protein that promotes joint molecule formation and strand-transfer reactions in vitro. We show here that this activity is also expressed by vaccinia DNA polymerase (gpE9L). Recombinant vaccinia polymerase was produced using a hybrid vaccinia/T7 expression system and purified to homogeneity. This protein catalyzed joint molecule formation and strand transfer in vitro in reactions containing single-stranded circular and linear duplex DNAs. The reaction required homologous substrates and magnesium ions and was stimulated by DNA aggregating agents such as spermidine HCl and Escherichia coli single-strand DNA binding protein. There was no requirement for a nucleoside triphosphate cofactor. The reaction ceased when approximately 20% of the double-stranded substrate had been incorporated into joint molecules and required stoichiometric quantities of DNA polymerase (0.5-1 molecules of polymerase per double-stranded DNA end). Electron microscopy showed that the joint molecules formed during these reactions contained displaced strands and thus represented the products of a strand-exchange reaction. We also reexamined the link between replication and recombination using a luciferase-based transfection assay and cells infected with DNA polymerase Cts42 mutant viruses. These data substantiate the claim that there exists an inextricable link between replication and recombination in poxvirus-infected cells. Together, these biochemical and genetic data suggest a way of linking poxviral DNA replication with genetic recombination.

Catalysis↗
PubMed1999

Shope fibroma virus DNA topoisomerase catalyses holliday junction resolution and hairpin formation in vitro.

The telomeres of poxviral chromosomes comprise covalently closed hairpin structures bearing mismatched bases. These hairpins are formed as concatemeric replication intermediates and are processed into mature, unit-length genomes. The structural transitions and enzymes involved in telomere resolution are poorly understood. Here we show that the type I topoisomerase of Shope fibroma virus (SFV) can promote a recombination reaction which converts cloned SFV replication intermediates into hairpin-ended molecules resembling mature poxviral telomeres. Recombinant SFV topoisomerase linearised a palindromic plasmid bearing 1.5 kb of DNA encoding the SFV concatemer junction, at a site near the centre of inverted-repeat symmetry. Most of these linear reaction products bore hairpin tips as judged by denaturing gel electrophoresis. The resolution reaction required palindromic SFV DNA sequences and was inhibited by compounds which block branch migration (MgCl2) or poxviral topoisomerases. The resolution reaction was also slow, needed substantial quantities of topoisomerase, and required that the palindrome be extruded in a cruciform configuration. DNA cleavage experiments identified a pair of suitably oriented topoisomerase recognition sites, 90 bases from the centre of the cloned SFV terminal inverted repeat, which may mark the resolution site. These data suggest a resolution scheme in which branch migration of a Holliday junction through a site occupied by covalently bound topoisomerase molecules, could lead to telomere resolution.

DNA Topoisomerases, Type I↗