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Biomedical subjects

G O'Neill

Publications and source records attributed to G O'Neill.

At least 19 recordsLinked to original sources

Observer variation in the sonographic measurement of optic nerve sheath diameter in normal adults.

AIM: To quantify the observer variation in the sonographic measurement of optic nerve sheath diameter (ONSD) in normal adults. MATERIALS AND METHODS: Sixty-seven normal adult volunteers underwent ultrasound examination of each eye by three independent observers using a 7 MHz sector probe. Three measurements were made of each eye by each observer and a mean value calculated for each eye. Median and 5th-95th centile values for both intra- and inter-observer variation were calculated for the three observers. RESULTS: The median intra-observer variation was +/-0.1 mm, with 5th-95th centile values of +/-0-0.4 mm. The median inter-observer variation was +/-0.2-0.3 mm, with 5th-95th centile values of +/-0-0.7 mm. Careful review of examination technique by the three observers after the first 17 examinations was shown to reduce both intra- and inter-observer variation. CONCLUSION: The sonographic measurement of ONSD is a readily learned, reproducible technique with low intra- and inter-observer variation. The average inter-observer variation (+/-0.2 mm) is comparable to the inherent variability of the ultrasound machine. The importance of standardisation of examination technique is stressed.

Adult↗

The occurrence of Corynosoma strumosum in the grey seal, Halichoerus grypus, caught off the Atlantic coast of Ireland.

The occurrence, location and sex ratio of Corynosoma strumosum (Acanthoceophala: Polymorphidae) from 26 juvenile grey seals, Halichoerus grypus, by-caught from around the Inishkea Islands, Co. Mayo, from March to June 1997 were recorded. The location of C. strumosum within the seal did not vary, with all worms being found in the small intestine. The prevalence of infection was 100%, with no other acanthocephalan species being recorded. The mean abundance was 416 (range 80-846) C. strumosum per seal. There was no significant change in abundance by month. The sex ratio of mature female to mature male parasites was 1.2:1. The lengths of female and male worms did not differ significantly.

Acanthocephala↗

Frequent loss of estrogen receptor-beta expression in prostate cancer.

The role of estrogen and its receptors in the etiology and progression of prostate cancer (PC) is poorly understood. In normal and malignant human prostate, estrogen receptor-alpha is expressed only in the stroma, whereas estrogen receptor-beta (ERbeta) is present in both normal stroma and epithelium. Because loss of ERbeta expression is associated with prostate hyperplasia in ERbeta-null mice, this study determined patterns of ERbeta expression in normal, hyperplastic, and malignant human prostate and associations with clinical outcome. Five normal prostates from organ donors and 159 radical prostatectomy specimens from patients with clinically localized PC were assessed for ERbeta expression using immunohistochemistry. ERbeta-positivity was defined as > or =5% of cells demonstrating nuclear immunoreactivity. All of the five normal prostates showed strong ERbeta-nuclear staining in >95% of the epithelium and 35% of the stromal cells. The number of ERbeta-positive cases declined to 24.2% (38/157) in hyperplasia adjacent to carcinoma and 11.3% (18/159) in PCs. ERbeta-positivity was related to decreased relapse-free survival (log-rank P = 0.04). Thus, loss of ERbeta expression is associated with progression from normal prostate epithelium to PC, whereas those cancers that retained ERbeta expression were associated with a higher rate of recurrence. These data identify the need to further investigate the potential role of ERbeta in the regulation of prostate epithelial cell proliferation and the functional consequences of decreased ERbeta expression in the evolution of PC.

Adult↗

Dual AV nodal pathway physiology after injury with radiofrequency energy in a patient without a history of reentrant tachycardia.

Radiofrequency (RF) atrioventricular (AV) nodal modification has been reported to occasionally produce a proarrhythmic effect. Dual AV nodal pathway physiology in patients without reentrant tachychardia has also been reported. This case describes AV nodal modification with RF energy in an anatomically intermediate area resulting in the appearance of discontinuous antegrade conduction curves and reentry in a patient in which these were previously not present. This suggests that AV nodal injury may be a mechanism for acquired AV nodal reentry.

Atrial Fibrillation↗

Methadone and the pregnant user: a matter for careful clinical consideration.

The practice of placing all pregnant heroin users on methadone as the treatment of choice needs to be questioned. While it may be suitable for those who stabilise their illicit heroin use at or shortly after conception, its suitability for those who show little movement away from regular heroin use and its associated lifestyle is more circumspect. Neonates of women who continue heroin use throughout pregnancy are likely to be below birthweight and/or premature. As a consequence, they are in a less than optimal condition to cope with the additional assault caused by prescribed methadone such as neonatal withdrawal. This may help explain why the relative risk of neonatal mortality in women who continue illicit heroin use during pregnancy and are prescribed methadone, is greater than for those who continue to use heroin but are not prescribed methadone. Clinicians must take the time of maternal presentation and the likelihood of continued maternal heroin use into consideration when determining who is suitable for methadone and whether a reduced level of methadone will suffice. A number of different clinical scenarios are identified and possible management strategies discussed. The need to develop innovative services appropriate for pregnant women who continue regular heroin use, and for well-designed studies that define best practice for the management of these women is evident.

Female↗

Obstetric and neonatal outcomes associated with maternal naltrexone exposure.

Poor maternal and neonatal outcomes are associated with the pregnant heroin user. These include increased antepartum haemorrhage, decreased neonatal birthweight and increased neonatal mortality Medically supervised withdrawal from heroin during pregnancy has, however, been discouraged due to possible risk to the fetus and because of the high incidence of return to regular illicit heroin use by the mother. In recent years, however, a number of withdrawal procedures using anaesthesia, oral sedation, or intravenous sedation, precipitated by naloxone and/or naltrexone have been developed and carried out successfully on pregnant heroin users. We have now collated information on 18 cases (19 detoxifications) from three countries (Portugal, Australia and the United Kingdom). These case study data, although limited, indicate that detoxification of the pregnant heroin user is possible without significant risk to the neonate or mother, with many women not returning to dependent heroin use following detoxification. Naltrexone maintenance has also been used in the non-pregnant heroin user to discourage illicit heroin use. Similarly to methadone, stabilisation on naltrexone may be associated with conception and pregnancy Over the past three years, 26 women have conceived while on the Western Australia naltrexone program. Due to the unknown teratogenic effects, most have ceased naltrexone intake at approximately seven or eight weeks gestation. In a number of instances, however, naltrexone maintenance has been recommenced following return to a dependent pattern of heroin use. As a consequence, neonates have had different periods of naltrexone exposure, building from the initial seven or eight weeks. We now report on seven women who have delivered and three who are well into their third trimester. Neonatal and obstetric features were unremarkable with good Apgar scores, birthweight and head circumference observed. In the three cases still in third-term gestation, normal fetal development has been observed at recent ultrasound examinations. These case data indicate that naltrexone maintenance may have a role in the management of the pregnant heroin user.

Adult↗

Segregation of micrometer-dimension biosensor elements on a variety of substrate surfaces.

With the rapid development of micro total analysis systems and sensitive biosensing technologies, it is often desirable to immobilize biomolecules to small areas of surfaces other than silicon. To this end, photolithographic techniques were used to derivatize micrometer-sized, spatially segregated biosensing elements on several different substrate surfaces. Both an interference pattern and a dynamic confocal patterning apparatus were used to control the dimensions and positions of immobilized regions. In both of these methods, a UV laser was used to initiate attachment of a photoactive biotin molecule to the substrate surfaces. Once biotin was attached to a substrate, biotin/avidin/biotin chemistry was used to attach fluorescently labeled or nonlabeled avidin and biotinylated sensing elements such as biotinylated antibodies. Dimensions of 2-10 microm were achievable with these methods. A wide variety of materials, including glassy carbon, quartz, acrylic, polystyrene, acetonitrile-butadiene-styrene, polycarbonate, and poly(dimethylsiloxane), were used as substrates. Nitrene- and carbene-generating photolinkers were investigated to achieve the most homogeneous films. These techniques were applied to create a prototype microfluidic sensor device that was used to separate fluorescently labeled secondary antibodies.

Biosensing Techniques↗

Fluorescent amplified-fragment length polymorphism analysis of the MRSA epidemic.

Genotypes and putative genetic relationships were characterised for epidemic methicillin-resistant Staphylococcus aureus (EMRSA) strains and isolates from England and Wales, using a high resolution DNA fingerprinting technique, fluorescent amplified-fragment length polymorphism (FAFLP). Each of the phage types of EMRSA had a distinct FAFLP profile. The technique revealed clusters of strains and isolates, and could distinguish isolates belonging to the same phage type. FAFLP provides a new approach to the epidemiological study and control of MRSA.

DNA Fingerprinting↗

Sheep and goats: natural and experimental TSEs and factors influencing incidence of disease.

The major factor influencing incidence of disease following challenge with transmissible spongiform encephalopathies (TSEs) in sheep is the allotype at amino acid numbers 136, 154 and 171 of the PrP protein. There are at least two groups of TSEs, one which targets the amino acid encoded at position 136 and the other which is more influenced by the amino acid at codon 171. Within these groups of TSE types, there may additionally be sub-types, as resistance to some, but not all, "136-type" TSEs can also be affected by the amino acid at codon 154. In goats, there are also PrP polymorphisms which apparently influence incubation period of TSE disease, however, this has not found to be true for cattle and BSE incidence. Sheep PrP amino acid codons 136, 154 and 171 do not explain everything about, for example, natural scrapie occurrence in sheep flocks, and attention is now turning to the flanking regions of the PrP gene looking for sequence differences in gene expression control motifs which may also have an influence on disease development. The sheep PrP gene produces two mRNAs in peripheral tissues, the result of alternative polyadenylation in the 3' untranslated region of the gene. Results from transfection assays of murine neuroblastoma cells with constructs expressing different regions of ovine PrP mRNA have revealed the presence of sequences in the 3' untranslated region that modulate protein synthesis and have therefore the potential to affect disease progression.

3' Untranslated Regions↗

Systematic errors in bone conduction audiometry.

Air and bone conduction audiometry was carried out on two separate groups of 12 normal hearing volunteers. One group was tested at St Mary's hospital, the other at Charing Cross hospital. The results from both centres showed evidence of a systematic error in bone conduction resulting in a pattern of 'notching' at 2 kHz. We argue that the effect is likely to be more apparent in conductive deafness and that a significant distortion of the audiogram occurs in about 17% of such cases. Since the problem appears not to be restricted to the centres involved in the study, we strongly recommend that the issue be addressed by the appropriate professional bodies.

Adult↗

The detection of enterotoxins and toxic shock syndrome toxin genes in Staphylococcus aureus by polymerase chain reaction.

A simple polymerase chain reaction (PCR)-based procedure was developed for the detection of fragments of staphylococcal enterotoxins (SEs) SEA, SEB, SEC, SED, SEE, SEG, SEH, and SEI together with the toxic shock syndrome toxin (TSST-1) genes of Staphylococcus aureus. One hundred and twenty-nine cultures of S. aureus were selected, 39 of which were recovered from 38 suspected staphylococcal food-poisoning incidents. The method was reproducible, and 32 different toxin genotypes were recognized. The presence of SE genes was associated with S. aureus strains reacting with phages in group III, and the TSST-1 gene with phages in group I. There was a 96% agreement between the PCR results for detection of SEA-D and TSST-1 as compared with a commercial reverse passive latex agglutination assay for the detection of SEs from cultures grown in vitro. Enterotoxin gene fragments were detected in S. aureus cultures recovered from 32 of the 38 suspected staphylococcal food poisoning incidents, and of these, 17 were associated with SEE, SEG, SEH, and SEI in the absence of SEA-D. Simple PCR procedures were also developed for the detection of SE directly in spiked food samples, and this was most successfully achieved in mushroom soup and ham. Detection was less successful in three types of cheese and in cream. SEA or SEB were detected by enzyme-linked immunosorbent assay in three food samples (two of which were associated with food poisoning incidents) naturally heavily contaminated with S. aureus: the appropriate SEA or SEB gene fragments were detected directly in these three foods by PCR.

Animals↗

Novel mutations in the gene encoding ATP-binding cassette 1 in four tangier disease kindreds.

Tangier disease (TD) is an autosomal co-dominant disorder in which homozygotes have a marked deficiency of high density lipoprotein (HDL) cholesterol and, in some cases, peripheral neuropathy and premature coronary heart disease (CHD). Homozygotes are further characterized by cholesteryl ester deposition in various tissues throughout the body, most notably in those of the reticuloendothelial system. Several studies have demonstrated that the excess lipid deposition in TD is due to defective apolipoprotein-mediated efflux of cellular cholesterol and phospholipids. Although much progress has been made in our understanding of the metabolic basis of TD, the precise molecular defect had remained elusive until very recently. By positional cloning methods, we: 1) confirm the assignment of TD to chromosome 9q31, 2) provide evidence that human ATP-binding cassette-1 (hABC-1) maps to a 250 kb region on 9q31, and 3) describe novel deletion, insertion, and missense mutations in the gene encoding hABC-1 in four unrelated TD kindreds. These results establish a causal role for mutations in hABC-1 in TD and indicate that this transporter has a critical function in the regulation of intracellular lipid trafficking that dramatically affects plasma HDL cholesterol levels.

ATP-Binding Cassette Transporters↗

Receptor for motilin identified in the human gastrointestinal system.

Motilin is a 22-amino acid peptide hormone expressed throughout the gastrointestinal (GI) tract of humans and other species. It affects gastric motility by stimulating interdigestive antrum and duodenal contractions. A heterotrimeric guanosine triphosphate-binding protein (G protein)-coupled receptor for motilin was isolated from human stomach, and its amino acid sequence was found to be 52 percent identical to the human receptor for growth hormone secretagogues. The macrolide antibiotic erythromycin also interacted with the cloned motilin receptor, providing a molecular basis for its effects on the human GI tract. The motilin receptor is expressed in enteric neurons of the human duodenum and colon. Development of motilin receptor agonists and antagonists may be useful in the treatment of multiple disorders of GI motility.

Alternative Splicing↗

Synthesis of the nanomolar photoaffinity GABA(B) receptor ligand CGP 71872 reveals diversity in the tissue distribution of GABA(B) receptor forms.

A radioiodinated probe, [125I]-CGP 71872, containing an azido group that can be photoactivated, was synthesized and used to characterize GABA(B) receptors. Photoaffinity labeling experiments using crude membranes prepared from rat brain revealed two predominant ligand binding species at approximately 130 and approximately 100 kDa believed to represent the long (GABA(B)R1a) and short (GABA(B)R1b) forms of the receptor. Indeed, these ligand binding proteins were immunoprecipitated using a GABA(B) receptor-specific antibody confirming the receptor specificity of the photoaffinity probe. Most convincingly, [125I]-CGP 71872 binding was competitively inhibited in a dose-dependent manner by cold CGP 71872, GABA, saclofen, (-)-baclofen, (+)-baclofen and (L)-glutamic acid with a rank order and stereospecificity characteristic of the GABA(B) receptor. Photoaffinity labeling experiments revealed that the recombinant GABA(B)R2 receptor does not bind [125I]-CGP 71872, providing surprising and direct evidence that CGP 71872 is a GABA(B)R1 selective antagonist. Photoaffinity labeling experiments using rat tissues showed that both GABA(B)R1a and GABA(B)R1b are co-expressed in the brain, spinal cord, stomach and testis, but only the short GABA(B)R1b receptor form was detected in kidney and liver whereas the long GABA(B)R1a form was selectively expressed in the adrenal gland, pituitary, spleen and prostate. We report herein the synthesis and biochemical characterization of the nanomolar affinity [125I]-CGP 71872 and CGP 71872 GABA(B)R1 ligands, and differential tissue expression of the long GABA(B)R1a and short GABA(B)R1b receptor forms in rat and dog.

Affinity Labels↗

The laryngeal mask airway in tonsillectomy: the surgeon's perspective.

The laryngeal mask airway presents certain advantages in the management of the airway during general anaesthesia. However, from the surgeon's perspective, there have been reports of problems occurring when the laryngeal mask airway is used in tonsillectomy. This study of 90 patients undergoing tonsillectomy suggests that surgical access is inferior with a laryngeal mask airway and the weight of tonsillar tissue excised is less. In addition the laryngeal mask airway needs to be changed to an endotracheal tube during the procedure in 11.4% of patients.

Airway Obstruction↗

Genotyping of epidemic methicillin-resistant Staphylococcus aureus phage type 15 isolates by fluorescent amplified-fragment length polymorphism analysis.

Fluorescent amplified-fragment length polymorphism (FAFLP) analysis was investigated for its ability to identify and subtype isolates of an epidemic methicillin-resistant phage type of Staphylococcus aureus, EMRSA-15. These isolates were also characterized by PCR-restriction fragment length polymorphism (PCR-RFLP) of the coagulase gene and pulsed-field gel electrophoresis (PFGE). For FAFLP, DNA was double digested with restriction enzymes ApaI plus TaqI or EcoRI plus MseI. Site-specific adaptors were ligated to one or the other set of restriction fragments, and PCR amplification was carried out with adaptor-specific primers. Amplified fragments separated on an ABI 377 automated sequencer and analyzed with Genescan version 2.1 software generated FAFLP profiles for all the isolates. The presence or absence of fragments was scored, similarity coefficients were calculated, and UPGMA (unweighted pair group method using arithmatic averages) cluster analysis was performed. Either enzyme-primer combination readily differentiated EMRSA-15 from other methicillin-resistant S. aureus (MRSA) isolates and also revealed heterogeneity within the phage type. The discriminatory power of FAFLP was high. By combining both enzyme-primer data sets, 24 isolates were divided into 11 profiles. PCR-RFLP did not discriminate among these EMRSA-15 isolates. PFGE could discriminate well between isolates but was not as reproducible as FAFLP. All S. aureus and MRSA isolates in this study were typeable by FAFLP, which was easy to perform, robust, and reproducible, with evident potential to subtype MRSA for purposes of hospital infection control.

Bacteriophage Typing↗