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P O'Neill

Publications and source records attributed to P O'Neill.

At least 19 recordsLinked to original sources
PubMed1999

Quantitative modelling of DNA damage using Monte Carlo track structure method.

This paper presents data on modelling of DNA damage induced by electrons, protons and alpha-particles to provide an insight into factors which determine the biological effectiveness of radiations of high and low linear energy transfer (LET). These data include the yield of single- and double-strand breaks (ssb, dsb) and base damage in a cellular environment. We obtain a ratio of 4-15 for ssb:dsb for solid and cellular DNA and a preliminary ratio of about 2 for base damage to strand breakage. Data are also given on specific characteristics of damage at the DNA level in the form of clustered damage of varying complexity, that challenge the repair processes and if not processed adequately could lead to the observed biological effects. It is shown that nearly 30% of dsb are of complex form for low-LET radiation, solely by virtue of additional breaks, rising to about 70% for high-LET radiation. Inclusion of base damage increases the complex proportion to about 60% and 90% for low- and high-LET radiation, respectively. The data show a twofold increase in frequencies of complex dsb from low-LET radiation when base damage is taken into account. It is shown that most ssb induced by high-LET radiation have associated base damages, and also a substantial proportion is induced by low-energy electrons.

DNA↗
PubMed1999

DNA-PK activation by ionizing radiation-induced DNA single-strand breaks.

PURPOSE: To assess the ability of 60Co gamma-radiation-induced plasmid DNA single-strand breaks (gamma-ssb) to activate the DNA-dependent protein kinase (DNA-PK) in vitro. MATERIALS AND METHODS: Plasmid DNA was gamma-irradiated under aerobic conditions to yield 0-6 gamma-ssb and <0.1 double-strand breaks (dsb) per plasmid molecule. The irradiated DNA was used to stimulate DNA-PK in crude HF19 fibroblast nuclear extracts and/or purified HeLa cell DNA-PK protein, and the activation compared with that obtained with a single enzymatically generated plasmid DNA ssb (GpII endonuclease) or dsb (EcoRI endonuclease). RESULTS: Gamma-Irradiated plasmid DNA activates DNA-PK in both crude and purified preparations and the kinase activity increases linearly with dose. As significant DNA-PK activation was detectable using irradiated plasmids which contain <0.1 dsb/molecule, it was concluded that this activation is due to gamma-ssb. However, using purified DNA-PK, this activation is relatively weak as approximately 3 approximately-ssb is equivalent to one GpII-generated DNA ssb or one end of an EcoRI-generated dsb in DNA-PK assays. CONCLUSIONS: As gamma-ssb are in a approximately 20-fold excess of approximately-dsb in vivo for low LET radiation, gamma-ssb may contribute significantly to DNA-PK signalling of gamma-radiation-induced DNA damage in vivo.

Cells, Cultured↗
PubMed1998

Guanine is the target for direct ionisation damage in DNA, as detected using excision enzymes.

Exposure of an aqueous, aerated solution (pH 7) of a double-stranded DNA to 193 nm light, of sufficient energy to ionise DNA, leads to selective, non-random modification at guanine in the form of frank single-strand break (ssb) and base modifications, revealed by treatment with either Escherichia coli formamidopyrimidine-DNA glycosylase (Fpg), Escherichia coli endonuclease III (Nth) or hot piperidine treatment. There is a similar neighbouring base sequence dependence for Fpg- and Nth-sensitive damage as that previously reported for both hot alkali-labile damage and prompt ssb. Low yields of photoproducts, namely pyrimidine dimers, are also revealed using the enzyme T4 endonuclease V (T4 endo V). Although irradiation of DNA with 193 nm light causes photoionisation of all the nucleic acid bases, these results indicate that guanine is the predominant site for localisation of the oxidative damage. These findings are consistent with migration of the radical cation to 'target' damage at guanine sites.

Base Sequence↗
PubMed1998

Cu,Zn superoxide dismutase from Photobacterium leiognathi is an hyperefficient enzyme.

The catalytic rate constant of recombinant Photobacterium leiognathi Cu,Zn superoxide dismutase has been determined as a function of pH by pulse radiolysis. At pH 7 and low ionic strength (I = 0.02 M) the catalytic rate constant is 8.5 x 10(9) M-1 s-1, more than two times the value found for all the native eukaryotic Cu,Zn superoxide dismutases investigated to date. Similarly, Brownian dynamics simulations indicate an enzyme-substrate association rate more than two times higher than that found for bovine Cu,Zn superoxide dismutase. Titration of the paramagnetic contribution to the water proton relaxation rate of the P. leiognathi with increasing concentration of halide ions with different radii indicates that the proteic channel delimiting the active site is wider than 4.4 A. This is at variance with that found on the eukariotic enzymes, and provides a rationale for the high catalytic rate of the bacterial enzyme. Evidence for solvent exposure of the active site different from that observed in the eukaryotic enzyme is suggested from the pH dependence of the water proton relaxation rate and of the EPR spectrum line shape, which indicate the occurrence of a prototropic equilibrium at pH 9.1 and 9.0, respectively. The pH dependence of the P. leiognathi catalytic rate has a trend different from that observed in the bovine enzyme, indicating that groups differently exposed to the solvent are involved in the modulation of the enzyme-substrate encounter.

Animals↗
PubMed1998

Conserved enzyme-substrate electrostatic attraction in prokaryotic Cu,Zn superoxide dismutases.

The catalytic activity of wild type Escherichia coli Cu,Zn superoxide dismutases and of two mutants in which two lysine residues conserved in most bacterial Cu,Zn superoxide dismutases have been replaced by serine was investigated by pulse radiolysis and Brownian dynamics simulations. Experimental and computational data show that neutralization of Lys60 strongly reduces the catalytic activity of the enzyme (approximately 50%), indicating that this residue has a primary role in the electrostatic attraction of the substrate towards the catalytic copper. Neutralization of Lys63 does not significantly influence the catalytic rate constant. The results suggest that prokaryotic Cu,Zn superoxide dismutases have evolved an electrostatic mechanism to facilitate the enzyme-substrate encounter that is functionally equivalent to that already found in the eukaryotic enzymes.

Amino Acid Sequence↗
PubMed1998

Does post-traumatic stress disorder occur after stroke: a preliminary study.

BACKGROUND AND PURPOSE: Psychological reactions after stroke have been recognized for some time. The present study examined whether psychological symptoms consistent with post-traumatic stress disorder (PTSD) could occur after stroke as a consequence of the sudden and unpredictable occurrence of a life-threatening internal stressor. METHODS: Sixty-one patients who had experienced a first-ever stroke or transient ischaemic attack were assessed using standard self-report clinical measures for anxiety, depression and PTSD. Those patients who fulfilled criteria on at least one PTSD self-report measure were subsequently assessed using a structured clinical interview for PTSD. RESULTS: Six (9.8%) patients fulfilled criteria for PTSD. No significant differences were found between the post-stroke PTSD group and the non-PTSD group in terms of premorbid health and lifestyle, or experiences of adverse life events. Significant differences were noted with respect to self-reported post-stroke mental health and premorbid neuroticism. The post-stroke PTSD group also scored consistently higher on measures of anxiety, depression and psychiatric caseness. CONCLUSIONS: The results of this study demonstrate a number of close similarities between post-stroke PTSD and classical PTSD. We therefore conclude that PTSD or a PTSD-like syndrome can occur after stroke.

Adult↗
PubMed1998

Role of the electrostatic loop charged residues in Cu,Zn superoxide dismutase.

We have expressed and characterized a mutant of Xenopus laevis Cu,Zn superoxide dismutase in which four highly conserved charged residues belonging to the electrostatic loop have been replaced by neutral side chains: Lys120 --> Leu, Asp130 --> Gln, Glu131 --> Gln, and Lys134 --> Thr. At low ionic strength, the mutant enzyme is one of the fastest superoxide dismutases ever assayed (k = 6.7 x 10(9) M(-1) s(-1), at pH 7 and mu = 0.02 M). Brownian dynamics simulations give rise to identical enzyme-substrate association rates for both wild-type and mutant enzymes, ruling out the possibility that enhancement of the activity is due to pure electrostatic factors. Comparative analysis of the experimental catalytic rate of the quadruple and single mutants reveals the nonadditivity of the mutation effects, indicating that the hyperefficiency of the mutant is due to a decrease of the energy barrier and/or to an alternative pathway for the diffusion of superoxide within the active site channel. At physiological ionic strength the catalytic rate of the mutant at neutral pH is similar to that of the wild-type enzyme as it is to the catalytic rate pH dependence. Moreover, mutation effects are additive. These results show that, at physiological salt conditions, electrostatic loop charged residues do not influence the diffusion pathway of the substrate and, if concomitantly neutralized, are not essential for high catalytic efficiency of the enzyme, pointing out the role of the metal cluster and of the invariant Arg141 in determining the local electrostatic forces facilitating the diffusion of the substrate towards the active site.

Animals↗
PubMed1998

The effect of body composition on hexarelin-induced growth hormone release in normal elderly subjects.

OBJECTIVE: Growth hormone (GH) release is influenced by several factors including age, gender, physical exercise, nutritional status, sex steroids and body composition. The relationship with body composition is complex. Obesity is accompanied by suppression of spontaneous and stimulated GH release. As increasing body fat reduces stimulated GH secretion following a standard provocative test, the potential clinical uses of GH-releasing peptides (GHRPs), therapeutically or diagnostically, may be dependent on the relationship between body fat and GHRP-stimulated GH release. We have therefore assessed the effect of body composition and gender on the GH releasing capacity of hexarelin. DESIGN: A single bolus of subcutaneous hexarelin at a dose of 1.5 micrograms per kg of body weight was administered at time 0. Blood samples were taken at -10, 0, 10, 20, 30, 40, 50, 60, 90, 120, 170 and 180 min. SUBJECTS: Twenty-one (eight male) healthy elderly subjects with a median (range) age of 68 (60-81) years and BMI of 26 (19-30) kg/m2 were studied. METHODS: Dual-energy X-ray absorptiometry (DEXA) was used to assess body composition. RESULTS: Peak GH response correlated negatively with fat mass, BMI, percentage body fat, and weight [r = -0.72, P = 0.0001; r = -0.56, P = 0.009; r = -0.63, P = 0.002 and r = -0.48, P = 0.029, respectively,]. AUC GH correlated negatively with fat mass, BMI and percentage fat mass [r = -0.58, P = 0.006; r = -0.51, P = 0.019 and r = -0.66, P = 0.001 respectively]. Using multiple linear regression, fat mass was the most useful predictor for both peak GH response [R2 = 0.61, P < 0.0001] and AUC GH [R2 = 0.38, P = 0.003]. Gender was not a significant variable. CONCLUSIONS: Increasing total fat mass results in a blunted GH response following subcutaneous hexarelin. Total fat mass appears to be a useful predictor of peak GH response even in normal individuals as none of the subjects in the present study was morbidly obese. This indicates that there is a continuum of effect of fat mass on hexarelin-stimulated GH release. Any impact of gender on the GH response to hexarelin is almost certainly indirect and mediated via differences in body composition. This observation will have an impact on the potential diagnostic and therapeutic uses of hexarelin and related GH secretagogues.

Aged↗
PubMed1998

Why are there eccentricity effects in visual search? Visual and attentional hypotheses.

In standard visual search experiments, observers search for a target item among distracting items. The locations of target items are generally random within the display and ignored as a factor in data analysis. Previous work has shown that targets presented near fixation are, in fact, found more efficiently than are targets presented at more peripheral locations. This paper proposes that the primary cause of this "eccentricity effect" (Carrasco, Evert, Chang, & Katz, 1995) is an attentional bias that allocates attention preferentially to central items. The first four experiments dealt with the possibility that visual, and not attentional, factors underlie the eccentricity effect. They showed that the eccentricity effect cannot be accounted for by the peripheral reduction in visual sensitivity, peripheral crowding, or cortical magnification. Experiment 5 tested the attention allocation model and also showed that RT x set size effects can be independent of eccentricity effects. Experiment 6 showed that the effective set size in a search task depends, in part, on the eccentricity of the target because observers search from fixation outward.

Adolescent↗
PubMed1998

Laser line-scanning confocal fluorescence imaging of the photodynamic action of aluminum and zinc phthalocyanines in V79-4 Chinese hamster fibroblasts.

Confocal fluorescence microscopy, using a newly constructed laser line-scanning confocal microscope, was applied to an investigation of the early stages of photoinduced destruction of V79-4 Chinese hamster fibroblasts using aluminum and zinc phthalocyanines as photosensitizers. Results obtained in this work show that aluminum and zinc phthalocyanines, once internalized, localize in perinuclear sites that are disrupted upon light exposure resulting in fluorescence redistribution. The combination of laser-line scanning with charge-coupled device detection used in the confocal microscope developed in this work can enable rapid high-resolution sequential imaging, which is ideal for studying photoinduced intracellular fluorescence dynamics.

Animals↗
PubMed1998

Induction of DNA-protein crosslinks in Chinese hamster V79-4 cells exposed to high- and low-linear energy transfer radiation.

The induction of DNA-protein crosslinks (DPCs) in Chinese hamster V79-4 cells after irradiation under hypoxic and aerobic conditions at 277 K with 60Co gamma rays, 238Pu alpha particles and aluminum K (Al(K)) ultrasoft X rays has been determined using a nitrocellulose filter binding assay. The dose dependences for the induction of DPCs, which involves covalent linkage, are linear over the absorbed dose range used (0-400 Gy with alpha-particle and gamma radiation, 0-600 Gy with Al(K) X rays). The yield of DPCs induced under hypoxic conditions is 55, 51 and 25 DPCs per gray per cell for 60Co gamma rays, alpha particles and Al(K) X rays, respectively. The yield of DPCs is significantly reduced in the presence of oxygen by 20, 50 and 79% for 60Co gamma rays, alpha particles and Al(K) X rays, respectively. Since the mean size of the DNA attached to the protein is uniform for 60Co gamma rays and alpha particles, variations in the DNA size do not influence the yields of DPCs. Although a DPC may be considered as a complex lesion combining two macromolecules, the dependence of the yield of DPCs on LET does not reflect the ionizing density of the radiations used. Further, this dependence on LET and the effect of oxygen do not reflect the corresponding dependences determined for a variety of biological responses. From these findings and knowledge of the radiation tracks, it is proposed that DPCs induced particularly under aerobic conditions with 60Co gamma rays are formed mainly in the sparsely ionizing segments of the radiation track.

Animals↗
PubMed1997

Building a relationship: communications and relationships between staff and stroke patients on a rehabilitation ward.

Communications among staff and patients on a stroke rehabilitation ward form the focus of this article, which reports on some aspects of a larger study using a grounded theory approach. Tape-recorded interviews were transcribed and analysed concurrently according to recommendations for the approach. A main theme entitled building a relationship was identified, and this process was found to occur in a context varying from participative at one end of a continuum to hierarchical at the other. Building a relationship was found to be influenced by role, personal qualities and organizational context. Appropriate relationships between role-holders were subject to negotiation, leading to a resulting congruence or incongruence between participants' expectations of each other and their roles. Personal qualities were brought into play in the process, with patients' views of staff and staff views of patients both being influential. Some of these views seemed to parallel what has been described in earlier literature as 'the sick role' and the labelling of patients as 'good' or 'bad'. Responses to personal qualities led to nurses ascribing meaning to patients' behaviour in terms of adjustment to their stroke, giving time to them to help them to adjust, and withdrawal and handing over to other staff if this strategy failed. Organizational context also had an influence on building a relationship, with time constraints being identified particularly by nurses, and the need to fit in the most essential aspects of care. Place was also important, in that nurses were confined to the ward as a work location, whereas other therapists and doctors worked in other places and sometimes had the facility to take patients off the ward to concentrate on therapy. The findings are discussed against the background of related literature and the conclusion is drawn that the crucial role of nurses in rehabilitation is not recognized and valued, and that shortages of resources-especially suitably qualified and trained nursing staff-are a negative influence on building the relationships which are vital to successful rehabilitation.

Cerebrovascular Disorders↗
PubMed1997

Computational modelling of low-energy electron-induced DNA damage by early physical and chemical events.

Modelling and calculations are presented as a first step towards mechanistic interpretation and prediction of radiation effects based on the spectrum of initial DNA damage produced by low energy electrons (100 eV-4.5 keV) that can be compared with experimental information. Relative yields of single and clustered strand breaks are presented in terms of complexity and source of damage, either by direct energy deposition or by reaction of OH radicals, and dependence on the activation probability of OH radicals and the amount of energy required to give a single strand break (ssb). Data show that the majority of interactions in DNA do not lead to damage in the form of strand breaks and when they do occur, they are most frequently simple ssb. However, for double-strand breaks (dsb), a high proportion (approximately 30%) are of more complex forms, even without considering additional complexity from base damage. The greater contribution is from direct interactions in the DNA but reactions of OH radicals add substantially to this, both in terms of the total number of breaks and in increasing the complexity within a cluster. It has been shown that the lengths of damaged segments of DNA from individual electron tracks tend to be short, indicating that consequent deletion length (simply by loss of a fragment between nearby dsb) would be short, very seldom exceeding a few tens of base pairs.

Computer Simulation↗
PubMed1997

Interactions between transvenous nonthoracotomy cardioverter defibrillator systems and permanent transvenous endocardial pacemakers.

Limited information is available regarding potential adverse interactions between transvenous nonthoracotomy cardioverter defibrillators and pacemakers. We describe our experience with 37 patients who have undergone successful implantation of both a transvenous defibrillator and pacemaker. The patients' mean age was 64 +/- 12.9 years. Thirty-three were male and four were female. The mean LVEF was 30.8% +/- 11.8%. The indications for pacemaker implantation included sick sinus syndrome in 13 patients, complete heart block in 15 patients, sinus bradycardia secondary to medications in 8 patients, and neurocardiogenic syncope in 1 patient. The indications for insertion of a defibrillator included medically refractory VT in 27 patients and sudden cardiac death in 10 patients. Twenty-three patients received an Endotak lead and 14 patients received a Transvene lead. Eighteen patients had a pacemaker prior to an ICD, 14 patients had an ICD prior to a pacemaker, and 4 patients had both devices placed simultaneously. Interaction was evaluated at implant of the second device and 1-3 days after both devices were placed. Detection of VF/VT was analyzed during asynchronous pacing (DOO/VOO) with maximum pacing output. In addition, in six patients, DFT was determined before and after pacemaker implantation. In 14 patients (38%), device interactions that could not always be optimally corrected were observed. In five patients, the pacemaker was reset to the "noise reversion" mode after high energy ICD discharge. Oversensing of atrial pacemaker stimuli resulted in inappropriate ICD firings in four patients. This was observed only with a specific device and could not be prevented by atrial lead repositioning in two of them, but required reprogramming of the pacemaker to the VVI mode. An increase in DFT was observed in five patients who had a pacemaker implanted after an ICD. Compared with previously published studies, a greater frequency of transvenous ICD and pacemaker interactions were observed. Considering that almost 50% of the patients already have a pacemaker at the time of ICD implant, the availability of defibrillators with dual chamber pacing capability will not eliminate the potential for this problem.

Aged↗
PubMed1997

Irradiation of DNA with 193 nm light yields formamidopyrimidine-DNA glycosylase(Fpg) protein-sensitive lesions.

Irradiation of aqueous solutions of plasmid DNA (pUC18) at pH 7.6 with 193 nm laser light results in low yields of prompt single strand breakage (air-saturated sample phi ssh = [1.5 +/- 0.1] x 10(4), argon-saturated sample phi ssh = [0.9 +/- 0.1] x 10(4). Treatment of the irradiated DNA samples with Escherichia coli formamidopyrimidine-DNA glycosylase (Fpg) protein results in an approximate 20-fold increase in the yield of single strand break-age (air-saturated sample phi fpg = [33.1 +/- 3.1] x 10(-4), argon-saturated sample phi fpg = [23.8 +/- 2.6] x 10(-4). This result indicates that 193 nm light induces other modification(s) (most likely of the purine moieties) that are 20 times more abundant than prompt strand breakage within the DNA matrix.

DNA Adducts↗
PubMed1997

Induction and rejoining of DNA double-strand breaks in Chinese hamster V79-4 cells irradiated with characteristic aluminum K and copper L ultrasoft X rays.

Characteristic aluminum K (AlK) (energy of 1.5 keV) and copper L (CuL) (energy of approximately 0.96 keV) ultrasoft X rays have been used to investigate the effectiveness of the numerous low-energy secondary electrons produced by low-linear energy transfer (LET) ionizing radiation. Cellular inactivation and induction and rejoining of DNA double-strand breaks (DSBs) in Chinese hamster V79-4 cells irradiated as monolayers with these ultrasoft X radiations have been studied under aerobic and anaerobic conditions. The mean cell thickness, determined by confocal laser scanning fluorescence microscopy, was used to calculate the mean dose to the nucleus of the irradiated cells. Relative to 60Co gamma rays, the relative biological effectiveness (RBE) for cellular inactivation at 10% survival is 1.7 +/- 0.1 and 2.3 +/- 0.3 for AIK and CuL ultrasoft X rays, respectively. The RBE values for induction of DSBs of 2.5 +/- 0.2 and 3.0 +/- 0.3 for AlK and CuL X rays, respectively, were determined after irradiation at 277 K using the technique of pulsed-field gel electrophoresis. Induction of DSBs is linearly dependent on dose. Oxygen enhancement ratios of 1.9 and 2.1 for cellular inactivation and DSB induction, respectively, were obtained with AIK X rays. These values are less than those for 60Co gamma radiation. The repair kinetics for rejoining of DSBs after a dose of 15 Gy is similar for both X-ray energies and 60Co gamma rays with a first half-life of 18-22 +/- 5 min. From these studies, it is suggested that induction of DSBs by low-LET radiations such as 60Co gamma rays reflects clustered damage produced predominantly by low-energy electron "track ends," which represent about 30% of the total dose.

Aerobiosis↗