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Biomedical subjects

M Edwards

Publications and source records attributed to M Edwards.

At least 19 recordsLinked to original sources

Erythroid expression and DNAaseI-hypersensitive sites of the carbonic anhydrase 1 gene.

The carbonic anhydrase 1 gene is expressed in adult human and mouse erythroid cells and colon epithelia from two distinct promoters. We have explored the erythroid promoter for cis-acting sequences involved in transcription using DNAaseI as a probe. Two DNAaseI-hypersensitive sites (DHS-1 and DHS-2) have been identified in the distal erythroid promoter in CA1-expressing erythroleukaemic cells. These sites are present at low levels in K562 cells, which have a foetal/embryonic phenotype and do not express CA1. DHS-1 and DHS-2 are not present in non-erythroid cells, including colon cells, which express CA1 from the proximal colon promoter. DHS-1 and DHS-2 were also generated in an heterologous CA1 gene containing 5 kb of erythroid promoter sequence after transfection into erythroid cells, including K562 cells. These transfection studies showed that both this fragment, and an abbreviated 817 bp promoter fragment which contains only DHS-1, were sufficient to confer erythroid-specific expression to a reporter gene. These promoters were active in cell lines expressing CA1 and in K562 cells. This latter observation implies that a developmental repressor factor is both present in K562 cells and binds to a cis-acting sequence that is absent from the sequence 5 kb upstream of the erythroid transcription start site.

Base Sequence

Neural tube and other developmental anomalies in the guinea pig following maternal hyperthermia during early neural tube development.

Guinea pigs were exposed to hyperthermia for 1 hr once or twice on day 11, 12, 13, or 14 (E11-E14) of pregnancy. The mean rectal temperatures were elevated by 3.4 degrees C-4.0 degrees C. This treatment resulted in a marked elevation of rates of resorption and developmental defects in embryos examined at day E23. The defects observed were those affecting the neural tube (NTD) (exencephaly, encephaloceles, and microphthalmia), kyphosis/scoliosis, branchial arch defects, and pericardial edema. Embryos with NTD and kyphosis/scoliosis have not been found among newborn guinea pigs to date following maternal heat exposure on days E12-E14. It appears that embryos with these defects are filtered out by resorption or abortion by days E30-E35.

Abortion, Spontaneous

Gastroesophageal physiology after stapled gastroesophagostomy in dogs.

A stapled vertical gastroesophagostomy (VGE) has been proposed for benign esophageal stricture. The VGE involves performing an anastomosis using a linear cutting stapler inserted via a gastrotomy (one limb in the esophagus, the other in the proximal fundus). In phase I of the study, a 50-mm VGE was performed in 13 dogs; 7 received an additional fundoplication. Endoscopy with pinch biopsies, esophageal manometry, and 4-hour ambulatory pH recordings were performed in each dog prior to and 1 month after VGE. The only difference between groups postoperatively was a higher lower esophageal sphincter pressure in the group with the added fundoplication. The incidence of histologic esophagitis was low in both groups. In phase II of the study, a VGE was performed in six dogs using a 75-mm linear cutting stapler, accompanied by fundoplication in each dog. These dogs were then followed for 6 months to more completely assess the long-term risk of developing reflux esophagitis. At necropsy, no gross or histologic evidence of esophagitis was found. The performance of a VGE through the lower esophageal sphincter in normal dogs does not result in significant reflux esophagitis. VGE may be an alternative to resection for esophageal stricture.

Anastomosis, Surgical

Colour inputs to random-dot stereopsis.

Recently it has been claimed by Livingstone and Hubel that, of three anatomically and functionally distinct visual channels (the magnocellular, parvocellular interblob, and blob channels), only the magnocellular channel is involved in the processing of stereoscopic depth. Since the magnocellular system shows little overt colour opponency, the reported loss of the ability to resolve random-dot stereograms defined only by colour contrast seems consistent with this view. However, Julesz observed that reversed-contrast stereograms could be fused if correlated colour information was added. In the present study, 'noise' (non-corresponding) pixels were injected into random-dot stereograms in order to increase fusion time. All six subjects tested were able to achieve stereopsis in less than three minutes when there was only correspondence in colour and not in luminance, and three when luminance contrast was completely reversed. This ability depends on information about the direction of colour contrast, not just the presence of chromatic borders. When luminance and chromatic contrast are defined in terms of signal-to-noise ratios at the photoreceptor mosaic, chromatic information plays at least as important a role in stereopsis as does luminance information, suggesting that the magnocellular channel is not uniquely involved.

Color Perception

The human carbonic anhydrase I gene has two promoters with different tissue specificities.

Primer extension and S1 nuclease analysis of human carbonic anhydrase I (HCA1) mRNA transcripts in erythroid and non-erythroid tissues show that the HCA1 gene has two promoters with different tissue specificities, separated by 36 kb of DNA. The promoter of the HCA1 gene which is active in non-erythroid tissue shows strong sequence similarity with a similar region in the mouse CA1 gene, but has two equally used transcription start sites.

Animals

Structure and solution properties of tamarind-seed polysaccharide.

The major polysaccharide in tamarind seed is a galactoxyloglucan for which the ratios galactose:xylose:glucose are 1:2:25:2.8. A minor polysaccharide (2-3%) contains branched (1----5)-alpha-L-arabinofuranan and unbranched (1----4)-beta-D-galactopyranan features. Small-angle X-ray scattering experiments gave values for the cross-sectional radius of the polymer in aqueous solution that were typical of single-stranded molecules. Marked stiffness of the chain (C infinity 110) was deduced from static light-scattering studies and is ascribed partially to the restriction of the motion of the (1----4)-beta-D-glucan backbone by its extensive (approximately 80%) glycosylation. The rigidity of the polymer caused significant draining effects which heavily influenced the hydrodynamic behaviour. The dependence of "zero-shear" viscosity on concentration was used to characterise "dilute" and "semi-dilute" concentration regimes. The marked dependence on concentration in the "semi-dilute" region was similar to that for other stiff neutral polysaccharide systems, ascribed to "hyper-entanglements", and it is suggested that these may have arisen through a tenuous alignment of stiffened chains.

Carbohydrate Conformation

Physical mapping of the human carbonic anhydrase gene cluster on chromosome 8.

A cluster of genes encoding the three cytoplasmic carbonic anhydrase isozymes CAI, CAII, and CAIII lie on the long arm of chromosome 8 (8q22) in humans. These genes have been mapped using pulsed-field gel electrophoresis. The genes lie in the order CA2, CA3, CA1. CA2 and CA3 are separated by 20 kb and are transcribed in the same direction, away from CA1. CA1 is separated from CA3 by over 80 kb and is transcribed in the direction opposite to CA2 and CA3. The arrangement of the genes is consistent with proposals that the duplication event which gave rise to CA1 predated the duplication which gave rise to CA2 and CA3. The order of these three genes differs from that suggested for the mouse based on recombination frequency.

Blotting, Southern

Development of radiation protection standards.

Radiation protection standards are based on the best available knowledge, caution, and perception. Dose limits for occupational exposure have decreased as knowledge was gained about radiation effects: from 0.6 Sv (60 rem)/year for 1900-1930 to 50 mSv (5 rem)/year in 1958 (the level still used as of 1990). Current dose limits for public exposure range from 1 mSv to 5 mSv, depending on frequency of exposure. For the embryo and fetus, dose limits are 0.5 mSv/mo and 5 mSv for the entire gestation. In the 1970s, the concept of acceptable risk and that of a non-threshold dose-response relationship became the basis for setting dose limits. Three principles of radiation protection are that (a) dose levels should not exceed acceptable levels, (b) optimal dose levels should be as low as reasonably achievable, and (c) radiation should not be used unless it produces a positive net benefit. Although no dose limits have been set for patients undergoing diagnostic and therapeutic radiologic procedures, such measures must provide a net benefit to patients at optimal dose levels.

History, 20th Century

Cloning and characterization of a gene encoding pig epidermal growth factor.

A portion of the pig epidermal growth factor (EGF) gene has been isolated and characterized. The nucleotide sequencies of exons 20 and 21, which encode the EGF region of the precursor protein, show 85% similarity with the human EGF gene sequence. In addition, conservation of the intron-exon boundaries between the two species was generally observed. Although the pig exon 21 appeared to lack a single nucleotide at its 5' end relative to the human gene, sequences obtained by direct amplification of the genomic DNA around the 5' end of this exon using the polymerase chain reaction, and from a pig EGF cDNA recombinant isolated from a kidney library, indicated that the deletion was probably a cloning artifact. Comparison of the predicted amino acid sequence of pig EGF with that of EGF from other species, as well as with several other polypeptides which bind to the EGF receptor, indicated conservation of Gly18, Tyr37, Gly39 and Arg41 in addition to all six cysteine residues and Leu47, which are known to be critical for biological activity. A synthetic gene encoding the predicted amino acid sequence of pig EGF was expressed in yeast. The recombinant polypeptide was shown to compete with 125I-labelled mouse EGF for binding to cells and to stimulate DNA synthesis in quiescent monolayers of Swiss 3T3 cells.

Amino Acid Sequence

Airshow disaster plans.

Airshows require careful medical planning. Disaster drills and coordination between all rescue agencies are necessary. Rescue personnel and command post must be highly visible. Good communication includes dedicated radio frequencies at the site, dedicated phone lines to hospitals, and multiple back-ups. Security personnel should use yellow police tape for crowd control, and volunteers must be supervised. Key medical elements include efficient triage and stabilization at the site, adequate medical personnel, equipment dispersed within the crowd, rapid transportation including helicopters, and crisis intervention. Separate areas are needed for persons with minor injuries, relatives and friends, the press, and a morgue. Disaster plans for airshows have unique aspects. A predetermined time benefits planning. Airports often have pre-existing medical facilities, and participating military agencies may have additional resources. Burns and inhalation injuries predominate at airshow accidents. Enlarging spectator safety zones and modifying aerobatic maneuvers can prevent injury.

Aerospace Medicine

Autosomal recessive inheritance of myopia in Hong Kong Chinese infants.

Hong Kong Chinese infants were refracted by retinoscopy at regular intervals between the age of approximately 10 and 40 weeks. Cyclopentolate 1% was used to obtain cycloplegia. The spherical equivalent of the refractive error was calculated for each child and analysis of variance carried out according to the refractive status of the parents. From approximately 20 weeks of age a significant difference was found between the mean spherical equivalent of the refractive error of infants of parents both of whom were myopic and that of infants of parents neither of whom was myopic. At 40 weeks of age a good fit with an autosomal recessive probability model was obtained.

Adult

The refractive status of Hong Kong Chinese infants.

One hundred and fifty-eight full-term Hong Kong Chinese infants were examined, at the age of approximately 10 weeks, using retinoscopy. Fifty of the subjects were re-examined at the ages of 20, 30 and 40 weeks. Cyclopentolate 1% was used to produce cycloplegia. Photographic methods were used to assess the straightness of the eyes of 122 infants. While the mean spherical equivalent of the refractive error at 10 weeks was within the limits reported in studies of Caucasian infants, a rapid decrease, not previously reported, occurred between the ages of 10 and 40 weeks. The amounts of astigmatism found were in line with results from Caucasian eyes but the direction was overwhelmingly with-the-rule. The prevalence of anisometropia decreased with age and anisometropia was not prevalent at the age of 37 weeks. Two exotropes (1.6%) and no esotropes were identified although this may have been confounded by the use of cyclopentolate.

Astigmatism

Neuroimaging of AIDS.

A retrospective review of the neuroimaging procedures of 84 patients with the diagnosis of AIDS was performed. Both computed tomography (CT) and magnetic resonance imaging (MRI) procedures were evaluated for the presence of atrophy, enhancing lesions and focal non-enhancing lesions. The imaging findings in several infectious conditions (toxoplasmosis, cytomegalovirus, papovavirus, HIV virus, tuberculosis and histoplasmosis) are described. Intracranial lymphoma, another complication of AIDS, also is discussed.

Acquired Immunodeficiency Syndrome

Structure and methylation patterns of the gene encoding human carbonic anhydrase I.

The gene (CAI) encoding human carbonic anhydrase I (CAI) has been isolated and shown to have a total length of 50 kb. Some 36 kb of this consists of a large intron separating the erythroid-specific promoter from the coding region. A small (54 bp) noncoding exon from within this intron is occasionally found in transcripts. Two different polyadenylation sites have been found, the most distal of which is the most commonly used. Methylation levels near the promoter differ widely in cell lines. In CAI-expressing cells, a region of DNA near the promoter is demethylated in a generally highly methylated background. Surprisingly, non-CAI-expressing cell lines show much lower levels of methylation.

Amino Acid Sequence

Expression of a synthetic gene for horseradish peroxidase C in Escherichia coli and folding and activation of the recombinant enzyme with Ca2+ and heme.

A synthetic gene encoding horseradish peroxidase isoenzyme C (HRP C) has been synthesized and expressed in Escherichia coli. The nonglycosylated recombinant enzyme (HRP C*) was produced in inclusion bodies in an insoluble inactive form containing only traces of heme. HRP C* was solubilized and conditions under which it folded to give active enzyme were determined. Folding was shown to be critically dependent upon the concentrations of urea, Ca2+, and heme and on oxidation by oxidized glutathione. Purification of active HRP C* from the folding mixture gave a peroxidase, with about half the activity of HRP C. Glycosylation is thus not essential for correct folding and activity. The C-terminal and N-terminal extensions to HRP identified previously in cloned cDNA sequences are also not required for correct folding. However, Ca2+ appears to play a key role in folding to give the active enzyme. The overall yield of purified active enzyme was 2-3%, but this could be increased by reprocessing material that precipitated during folding.

Amino Acid Sequence