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Biomedical subjects

C Barry

Publications and source records attributed to C Barry.

At least 37 records · Page 2Linked to original sources
PubMed1996

Methylation of DNA repeats of decreasing sizes in Ascobolus immersus.

In Ascobolus immersus, DNA duplications are subject to the process of methylation induced premeiotically (MIP), which methylates the cytosine residues within the repeats and results in reversible gene silencing. The triggering of MIP requires pairing of the repeats, and its detection requires maintenance of the resulting methylation. MIP of kilobase-size duplications occurs frequently and leads to the methylation of all C residues in the repeats, including those belonging to non-CpG sequences. Using duplications of decreasing sizes, we observed that tandem repeats never escaped MIP when larger than 630 bp and showed a sudden and drastic drop in MIP frequencies when their sizes decreased from 630 to 317 bp. This contrasted with the progressive decrease of MIP frequencies observed with ectopic repeats, in which apparently the search for homology influences the MIP triggering efficiency. The minimal size actually required for a repeat to undergo detectable MIP was found to be close to 300 bp. Genomic sequencing and Southern hybridization analyses using restriction enzymes sensitive to C methylation showed a loss of methylation at non-CpG sites in short DNA segments, methylation being restricted to a limited number of CpG dinucleotides. Our data suggest the existence of two distinct mechanisms underlying methylation maintenance, one responsible for methylation at CpG sites and the other responsible for methylation at non-CpG sites.

Ascomycota↗
PubMed1996

Morphological organization of oligodendrocyte processes during development in culture and in vivo.

In order to meet the requirements for myelin biogenesis, the processes of oligodendrocytes must differentiate into specialized cellular compartments. For translation of myelin basic protein mRNA to occur in the oligodendrocyte processes or the myelin membrane, these structures must contain ribosomes and other components of the protein translation machinery. Light microscopy and electron microscopy (EM) were used to explore this possibility by analyzing the cytoskeletal organization and the organelle population of the processes of oligodendrocytes during development in vivo and in culture. Microtubules (MTs) were few and bundled in the younger processes of oligodendrocytes in culture. Some of these processes were characterized by the additional presence of serpentine MTs. EM revealed the presence of clusters of ribosomes in both immature and mature oligodendrocyte processes. In the former these clusters were frequently found at regular intervals along the processes. A similar interval characterized the distribution of myelin-like figures along the processes of mature cells in culture. Both in vivo and in culture, the processes became enriched in endoplasmic reticulum cisternae and mitochondria as the cells matured. The spatial arrangement of cellular organelles is compatible with protein and lipid synthesis occurring in the processes, at or near the site of myelin assembly.

Aging↗
PubMed1995

Physical and functional interaction of the Epstein-Barr virus BZLF1 transactivator with the retinoic acid receptors RAR alpha and RXR alpha.

Epstein-Barr virus (EBV) reactivation, indicated by induction of EBV early antigens from latently infected lymphoid cell lines by phorbol esters, is inhibited by retinoic acid (RA). Viral reactivation, which is triggered by the immediate-early BZLF-1 (Z) viral gene product, is repressed by retinoic acid receptors (RARs) RAR alpha and RXR alpha. These proteins negatively regulate Z-mediated transactivation of the promoter for an EBV early gene product, early antigen-diffuse (EaD). Here we confirm a direct physical interaction between the AP1-like protein Z and RXR alpha and map the domains of interaction in the Z protein and RXR alpha. The domain required for homodimerization of Z is separate from that required for its interaction with RXR alpha. Z also has the effect of repressing activation of an RAR-responsive cellular promoter (BRE). Point mutants in the dimerization domain of Z unable to interact with RXR alpha do not repress RXR alpha-mediated transactivation of BRE, the promoter for RAR beta, which suggests that interaction between the two proteins is required for this repressor effect. The domain of RXR alpha required for interaction with Z has been mapped, and is again separate from that required for homodimerization. These results indicate that a 'cross-coupling' or direct interaction between Z and RAR alpha and RXR alpha can modulate the reactivation of latent EBV infection and suggest that, reciprocally, the viral protein Z may influence cellular regulatory pathways.

Amino Acid Sequence↗
PubMed1995

Optimal conditions required for the creation of an iatrogenic chorioretinal venous anastomosis in the dog using argon green laser photocoagulation.

We have previously reported iatrogenic retinal to choroidal vein anastomosis, developed as a potential method of by-passing the site of obstruction to venous outflow in retinal venous occlusion in dogs (1), and in rats (2). In order to minimise tissue damage to the retina and choroid and increase the rate of success in these experiments, we investigated in the dog model the factors that would promote an anastomosis and compare the effects of three different power levels. A small spot size (50 microns) argon green laser beam of 514 nm at power levels of 0.5w, 1.5w and 2.5w were used. Spaced serial sections from each lesion were examined by light and by transmission electron microscopy. Morphometric measurements of the corresponding retinal pigment epithelium (RPE)/glial scar was computed using IBMPC digitising pad and sigma scan software and the extent of tissue damage at the different power levels assessed. At the lowest power level of 0.5w the damage to the retina was mild and there was an absence of anastomosis formation. At the 1.5w power level an anastomosis formed in 40% of the lesions. At the highest power level of 2.5w a 71% rate of success was obtained however, the damage to the retina tended to be severe. The results of this study also indicate that disruption of Bruch's membrane and vein rupture at the time of irradiation are essential for anastomosis formation, which may be further enhanced by necrotic tissue, retinal pigment epithelial and glial scar formation and inflammation. These findings are useful in establishing optimal conditions for the creation of a chorioretinal venous anastomosis, for consideration in human trials.

Anastomosis, Surgical↗
PubMed1994

Significance of Bruch's membrane in the creation of iatrogenic chorioretinal venous anastomosis.

Iatrogenic retinal to choroidal vein anastomoses, as a method of bypassing retinal venous occlusions has been reported in dogs in which Bruch's membrane is poorly formed. In order to determine whether chorioretinal venous anastomoses can be induced in an animal with a Bruch's membrane that is well developed as in humans, pigmented rats were chosen. A high intensity, small spot size argon green laser beam of 514 nm was used to induce the anastomosis. Three out of 5 rat eyes developed a retinal vein to choroidal vein anastomosis. The success rate of iatrogenic retinal to choroidal vein anastomoses in the rat was comparable to that obtained in the dog. This study suggests that Bruch's membrane was not significant in the creation of an iatrogenic chorioretinal venous anastomosis.

Anastomosis, Surgical↗
PubMed1993

Methylation induced premeiotically in Ascobolus: coextension with DNA repeat lengths and effect on transcript elongation.

The effect of duplications of gene fragments on the triggering of DNA methylation induced premeiotically (MIP) was studied in the sexual progeny of strains harboring, in addition to the resident met2 gene, a fragment of this gene inserted at an ectopic position. Cytosine methylation of the resident gene was checked for each of the eight duplications tested. Methylation was always found and it was coextensive with the length of the duplications. Silencing of the resident gene was triggered by duplications of segments corresponding to the region 5' to the open reading frame, to only the open reading frame, or to segments beginning 0.87-1.2 kb downstream from the transcription start. Silencing was accompanied by either the absence of transcripts or the presence of truncated transcripts, which suggests that methylation acts on transcript elongation.

5-Methylcytosine↗
PubMed1993

Performance of two films for densitometry of retinal photographs.

The relative performance was determined of two different photographic films (Kodak Panatomic-X and Kodak Technical Pan) for densitometry readings from retinal photographs of patients who had or were suspected of having glaucoma. The raw data from the two films were significantly different. The high contrast film was particularly sensitive to external variables; however, when normalised in terms of the standardised deviation, the data from the two films were comparable (P < 0.005). We also measured the pattern of nerve fibre layer loss using digital image analysis of red-free photographs of normal, ocular-hypertensive, glaucoma-suspect and glaucomatous eyes. Several mathematical techniques were used to characterise the data from each eye and then to compare these data to the mean photographic density of the normal eyes. Results showed that it was possible to separate normal eyes from glaucomatous eyes. The highest sensitivity achieved was 100% for right and 88% for left eyes, and the highest specificity was 100% for both eyes.

Adult↗
PubMed1993

Transport and localization of exogenous myelin basic protein mRNA microinjected into oligodendrocytes.

We have studied transport and localization of MBP mRNA in oligodendrocytes in culture by microinjecting labeled mRNA into living cells and analyzing the intracellular distribution of the injected RNA by confocal microscopy. Injected mRNA initially appears dispersed in the perikaryon. Within minutes, the RNA forms granules which, in the case of MBP mRNA, are transported down the processes to the periphery of the cell where the distribution again becomes dispersed. In situ hybridization shows that endogenous MBP mRNA in oligodendrocytes also appears as granules in the perikaryon and processes and dispersed in the peripheral membranes. The granules are not released by extraction with non-ionic detergent, indicating that they are associated with the cytoskeletal matrix. Three dimensional visualization indicates that MBP mRNA granules are often aligned in tracks along microtubules traversing the cytoplasm and processes. Several distinct patterns of granule movement are observed. Granules in the processes undergo sustained directional movement with a velocity of approximately 0.2 micron/s. Granules at branch points undergo oscillatory motion with a mean displacement of 0.1 micron/s. Granules in the periphery of the cell circulate randomly with a mean displacement of approximately 1 micron/s. The results are discussed in terms of a multi-step pathway for transport and localization of MBP mRNA in oligodendrocytes. This work represents the first characterization of intracellular movement of mRNA in living cells, and the first description of the role of RNA granules in transport and localization of mRNA in cells.

Animals↗
PubMed1992

Induced chorioretinal venous anastomosis in experimental retinal branch vein occlusion.

Iatrogenic retinal vein to choroidal vein anastomoses were created using laser photocoagulation in six of seven dog eyes in which a partial branch retinal vein occlusion had previously been created photochemically. A similar attempt to create an anastomosis was made in six control eyes in which no branch vein occlusion was present. In the eyes in which a branch retinal vein had been created, a venous chorioretinal anastomosis appeared to be present by 3 to 6 weeks. In three control eyes similar venous anastomosis was created; however this took 6 to 8 weeks to develop and was of much smaller calibre than the one that developed in the presence of a partial branch retinal vein occlusion. No adverse complications were noted in the period of the study (3 months). This study demonstrates that chorioretinal venous anastomoses can be created and may be of use in the treatment of partial retinal vein occlusions that show signs of progression.

Anastomosis, Surgical↗
PubMed1992

Intracellular pH in OK cells. II. Effects of temperature on cell pH.

Intracellular pH (pHi) is known to acidify as temperature rises. It has been proposed that this relationship reflects the temperature dependence of the dissociation constant of intracellular buffers. The purpose of this study was to evaluate whether H+ production and membrane acid/base transport events also contribute to the temperature dependence of pHi. This relationship was studied by means of the 490/450-nm fluorescence of 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein, to measure pHi of OK cells grown on cover slips and perfused in a thermostated chamber. As temperature was increased from 12 degrees C, pHi progressively acidified, until a precipitous and large alkalinization appeared at 40-45 degrees C. We specifically examined the range from 25 to 37 degrees C, over which pHi acidified by 0.32 units from 7.50 to 7.18. The three major components that regulate pHi in the OK cell were quantitated as follows: H+ production, H+ extrusion via Na(+)-H+ antiport, and the passive H+ leak from buffer to cell. Relative to the rate at 25 degrees C, acid production at 37 degrees C increased by 129% (3.82 +/- 0.78 vs. 8.76 +/- 1.12 mmol.l-1.min-1). The recovery of pHi after NH4Cl loading was used to measure the rate of Na(+)-H+ antiport, and in Na(+)-free depolarized cells the magnitude of passive H+ conductance through the leak pathway. Compared with the rate at 25 degrees C, at 37 degrees C the Na(+)-H+ antiport increased by 26% (1.25 +/- 0.22 vs. 1.58 +/- 0.25 mmol.l-1.min-1) and the H+ permeability increased by 140% (0.005 +/- 0.001 vs. 0.012 +/- 0.003 cm/s).(ABSTRACT TRUNCATED AT 250 WORDS)

Amiloride↗
PubMed1992

Avascular necrosis of the femoral head post heart-transplantation and steroid dosage.

Avascular necrosis (avn) post heart-transplantation has been considered to be due to the high doses of steroids used to immunosuppress these patients in attempting to prevent transplant rejection. This study shows that avascular necrosis occurs even when low dose steroids regimes are used and demonstrates no significant correlation between steroid dosage and the development of avn. Patients with symptomatic avn benefit from early diagnosis and management of their condition in that the need for total joint arthroplasty can be prevented in many cases.

Adult↗
PubMed1992

Low energy high frequency pulsed electromagnetic therapy for acute whiplash injuries. A double blind randomized controlled study.

The standard treatment of acute whiplash injuries (soft collar and analgesia) is frequently unsuccessful. Pulsed electromagnetic therapy PEMT (as pulsed 27 MHz) has been shown to have pro-healing and anti-inflammatory effects. This study examines the effect of PEMT on the acute whiplash syndrome. One half of the 40 patients entering the study received active PEMT collars: the other half facsimile (placebo). All patients were given instructions to wear the collar for eight hours a day at home and advised to mobilise their necks. At 2 and 4 weeks the actively treated group had significantly improved (p less than 0.05) in terms of pain (visual analogue scale). By chance movement scores for the PEMT group were significantly worse at entry to the study than the control group (p less than 0.05). At 12 weeks they had become significantly better (p less than 0.05). PEMT as described is safe for domiciliary use and this study suggests that PEMT has a beneficial effect in the management of the acute whiplash injury.

Accidents, Traffic↗