PubMed Health⌕ Search

Biomedical subjects

R D Saunders

Publications and source records attributed to R D Saunders.

At least 37 records · Page 2Linked to original sources

P-element insertion alleles of essential genes on the third chromosome of Drosophila melanogaster: correlation of physical and cytogenetic maps in chromosomal region 86E-87F.

We have established a collection of 2460 lethal or semi-lethal mutant lines using a procedure thought to insert single P elements into vital genes on the third chromosome of Drosophila melanogaster. More than 1200 randomly selected lines were examined by in situ hybridization and 90% found to contain single insertions at sites that mark 89% of all lettered subdivisions of the Bridges' map. A set of chromosomal deficiencies that collectively uncover approximately 25% of the euchromatin of chromosome 3 reveal lethal mutations in 468 lines corresponding to 145 complementation groups. We undertook a detailed analysis of the cytogenetic interval 86E-87F and identified 87 P-element-induced mutations falling into 38 complementation groups, 16 of which correspond to previously known genes. Twenty-one of these 38 complementation groups have at least one allele that has a P-element insertion at a position consistent with the cytogenetics of the locus. We have rescued P elements and flanking chromosomal sequences from the 86E-87F region in 35 lines with either lethal or genetically silent P insertions, and used these as probes to identify cosmids and P1 clones from the Drosophila genome projects. This has tied together the physical and genetic maps and has linked 44 previously identified cosmid contigs into seven "super-contigs" that span the interval. STS data for sequences flanking one side of the P-element insertions in 49 lines has identified insertions in the alphagamma element at 87C, two known transposable elements, and the open reading frames of seven putative single copy genes. These correspond to five known genes in this interval, and two genes identified by the homology of their predicted products to known proteins from other organisms.

Alleles↗

Acute exposure to power-frequency magnetic fields has no effect on the acquisition of a spatial learning task by adult male mice.

A series of four experiments was performed to determine whether acute exposure to a range of 50 Hz magnetic fields had any effect on a learning task in adult male CD1 mice. A radial-arm maze placed within the bore of an electromagnet was used to assess spatial discrimination learning for food reward. Subjects were reduced to 85% of their free-feeding weight and were placed in the maze for up to 15 minutes each day for 10 days. Performance of the task was measured by using maximum likelihood techniques to calculate the probability that an animal would not reenter any given arm of the maze. Experimental subjects were exposed to a vertical, 50 Hz sinusoidal magnetic field at 5 microT, 50 microT, 0.5 mT, or 5.0 mT (rms). Control subjects were exposed only to a background time-varying field of less than 50 nT and the ambient static field of about 40 microT. The variation in the applied magnetic field was less than 5% except at the ends of the arms, where it approached 10%. It was found that all eight groups of subjects (n = 10 in all cases) showed similar increases in performance with testing, and the acquisition curve for each group of experimental subjects was not significantly different from that of their control group (P > 0.05 in all cases). It was concluded that exposure had no effect on learning at any flux density. This result is contrary to the findings of a number of preliminary studies, although other studies have reported that magnetic fields do not affect spatial learning in adult male rodents. It is possible that differences between experimental conditions might explain some of this apparent discrepancy.

Animals↗

Prenatal exposure to a 50 Hz magnetic field has no effect on spatial learning in adult mice.

Male CD1 mice were exposed in utero to a 50 Hz sinusoidal magnetic field at 5 mT (rms) for the period of gestation and were raised subsequently without applied fields. At 82-84 days of age, they began a radial-arm-maze experiment that was designed to test for deficits in spatial learning and memory. Mice exposed in utero and sham-exposed mice exhibited no statistically significant differences in performances.

Animals↗

Effects of 50 Hz magnetic field exposure on the rate of DNA synthesis by normal human fibroblasts.

Interest in the potential adverse biological effects of exposure to power-frequency magnetic fields has centred on the possibility that these fields may influence tumour promotion, possibly by increasing the rate of cell proliferation. In order to investigate whether exposure to magnetic fields can indeed affect the rate of cell proliferation, normal human fibroblasts were serum starved overnight and then exposed to 50 Hz magnetic fields in a purpose-built facility. The rate of DNA synthesis was taken as a measure of cell proliferation, and was determined by following the incorporation of [3H]-thymidine into macromolecular material. The rate of DNA synthesis in exposed cells was compared with that in control cultures maintained in a standard CO2 incubator where they were exposed to background magnetic fields of < 200 nT. Positive controls were maintained in the same CO2 incubator, but were treated with human recombinant fibroblast growth factor to check that the cells were responsive to growth stimuli. Magnetic fields at 50 Hz and at a range of flux densities between 20 microT and 20 mT had no detectable effect on the rate of DNA synthesis by cells exposed for up to 30 h.

Cells, Cultured↗

Association of depression with reduced heart rate variability in coronary artery disease.

Decreased heart rate (HR) variability is an independent risk factor for mortality in cardiac populations. Clinical depression has also been associated with adverse outcomes in patients with coronary artery disease (CAD). This study tests the hypothesis that depressed patients with CAD have decreased HR variability compared with nondepressed CAD patients. Nineteen patients with angiographically documented CAD and either major or minor depression were compared with a sample of nondepressed CAD patients according to age, sex, and smoking status. All patients underwent 24-hour Holter monitoring, and the standard deviation of all normal-to-normal intervals was used as the primary index of HR variability. HR variability was significantly lower in depressed than nondepressed patients (90 +/- 35 vs 117 +/- 26 ms; p < or = 0.01), even after adjusting for relevant covariates. Thus, decreased HR variability may help explain the increased risk for cardiac mortality and morbidity in depressed CAD patients.

Aged↗

Dominant lethal studies in male mice after exposure to a 50 Hz magnetic field.

The potential mutagenicity of power frequency magnetic fields was investigated using a dominant lethal assay in mice. A total of 42 male mice were exposed for 8 weeks to a 50 Hz sinusoidal magnetic field at 10 mT (rms) and 47 males acted as simultaneous cage controls. Each male was subsequently mated with two females on weeks 1, 3, 5, 7, and 9 post-exposure. The numbers of pregnant females, corpora lutea, and live and dead implants were recorded. Multiple logistic regression analyses examined the effects of exposure on pregnancy rate, pre-implantation survival and post-implantation survival. There were no statistically significant differences in overall response between exposed and control groups, nor was there any significant effect of exposure in any post-exposure week. Thus, exposure to power frequency magnetic fields at 10 mT for the approximate period of spermatogenesis did not appear to induce dominant lethal mutation in the germ cells of male mice.

Animals↗

A physical map of the X chromosome of Drosophila melanogaster: cosmid contigs and sequence tagged sites.

A physical map of the euchromatic X chromosome of Drosophila melanogaster has been constructed by assembling contiguous arrays of cosmids that were selected by screening a library with DNA isolated from microamplified chromosomal divisions. This map, consisting of 893 cosmids, covers approximately 64% of the euchromatic part of the chromosome. In addition, 568 sequence tagged sites (STS), in aggregate representing 120 kb of sequenced DNA, were derived from selected cosmids. Most of these STSs, spaced at an average distance of approximately 35 kb along the euchromatic region of the chromosome, represent DNA tags that can be used as entry points to the fruitfly genome. Furthermore, 42 genes have been placed on the physical map, either through the hybridization of specific probes to the cosmids or through the fact that they were represented among the STSs. These provide a link between the physical and the genetic maps of D. melanogaster. Nine novel genes have been tentatively identified in Drosophila on the basis of matches between STS sequences and sequences from other species.

Amino Acid Sequence↗

Effects of prenatal exposure to 50 Hz magnetic fields on development in mice: I. Implantation rate and fetal development.

Pregnant CD1 mice were exposed or sham-exposed from day 0 to day 17 of gestation to a 50 Hz sinusoidal magnetic field at 20 mT (rms). Preimplantation and postimplantation survival were assessed and fetuses examined for the presence of gross external, internal, and skeletal abnormalities. There were no statistically significant field-dependent effects on preimplantation or postimplantation survival, sex ratio, or the incidence of fetuses with internal or skeletal abnormalities. Magnetic field exposure was, however, associated with longer and heavier fetuses at term, with fewer external abnormalities. The results lend no support to suggestions of increased rates of spontaneous abortion or congenital malformation following prenatal exposure to power frequency magnetic fields.

Abnormalities, Radiation-Induced↗

Effects of prenatal exposure to 50 Hz magnetic fields on development in mice: II. Postnatal development and behavior.

To investigate the potential of magnetic fields to act as a behavioral teratogen, pregnant CD1 mice were exposed or sham-exposed for all of gestation to a 50 Hz/20 mT magnetic field. Maturation of offspring was assessed using a range of standard developmental indices (eye opening, pinna detachment, hair coat, tooth eruption, sexual maturity, and weight) and simple reflexive behaviors (air righting, surface righting, forepaw grasp, cliff avoidance, and negative geotaxis). Activity and coordination levels were explored in juvenile and adult mice using an open field arena, a head-dip board, an accelerating Rotarod, and a residential activity wheel. All assessments were carried out without knowledge of exposure condition. Results from 168 sham-exposed mice from 21 litters and from 184 exposed mice from 23 litters were compared using survival analysis techniques and multivariate regression methods. Three possible field-dependent effects were found: Exposed animals performed the air righting reflex earlier (P < 0.01); exposed males (but not females) were significantly lighter in weight (P = 0.008) at 30 days of age; and exposed animals remained on a Rota-rod for less time as juveniles (P = 0.03). Some of these results have not been reported in other studies and may reflect spurious statistical significance, although some effect of magnetic field exposure cannot be ruled out. Overall, these results suggest that prenatal exposure to a 50 Hz magnetic field does not engender any gross impairments in the postnatal development or behavior of mice. This does not preclude such exposure affecting more subtle aspects of behavior.

Animals↗

Prenatal irradiation and spatial memory in mice: investigation of dose-response relationship.

Pregnant CD1 mice were exposed on gestational day 18 to 250 kV X-rays at 0.1, 0.25, 0.35 and 0.5 Gy. The performances of 10 adult male offspring from each exposure condition were investigated on a spatial discrimination learning task in a radial arm maze. An impairment in the performance of this task was found which showed a correlation with dose. Compared with sham exposed control mice, performance was not significantly affected with irradiation at 0.1 Gy and was slightly but non-significantly reduced at 0.25 Gy. Irradiation at 0.35 Gy caused a significant impairment in performance, and exposure at 0.5 Gy resulted in a still larger impairment. The overall association between dose and behavioural impairment was best described by a linear relationship without a threshold, although at doses lower than about 0.25 Gy any impairment would appear to be too small to be detectable.

Animals↗

Benzylamine antioxidants: relationship between structure, peroxyl radical scavenging, lipid peroxidation inhibition, and cytoprotection.

Three homologous series of 3,5-dialkoxy-4-hydroxybenzylamines were prepared and tested (1) as peroxyl radical scavengers in homogeneous aqueous solution, (2) as inhibitors of iron-dependent peroxidation of rabbit brain vesicular membrane lipids, and (3) as cytoprotective agents using primary cultures of rat hippocampal neurons exposed to hydrogen peroxide. The structural requirements for efficient radical trapping in homogeneous solution differed from those for effective lipid peroxidation inhibition: In homogeneous solution a kinetic preference existed for smaller, less sterically encumbered substituents flanking the reactive phenolic hydroxyl group. Lipid peroxidation inhibition, on the other hand, required longer more lipophilic substituents. Consequently, a lipophilic alkoxyl substituent at C3 and a small substituent at C5 appeared optimal for efficient radical scavenging activity in both lipid and homogeneous solution. Maximal cytoprotection of rat hippocampal neurons exposed to hydrogen peroxide was also associated with more lipophilic derivatives although substituent length and substituent bulk may represent independent parameters for relating structure and efficacy in this system.

Animals↗

Cloning of the fourth functional gene for protein phosphatase 1 in Drosophila melanogaster from its chromosomal location.

Complementary DNA encoding a catalytic subunit of protein phosphatase 1, PP1 87B, hybridises at four positions (87B, 9C, 13C and 96A) to Drosophila melanogaster polytene chromosomes, three of which are known to be expressed [Dombrádi, V., Axton, J.M., Brewis, N.D., Da Cruz e Silva, E.F., Alphey, L. & Cohen, P.T.W. (1990) Eur. J. Biochem. 194, 739-745]. The fourth gene has been isolated by screening a genomic library of cosmid clones, representing division 13 of the X-chromosome of D. melanogaster, with a PP1 87B probe. This library was constructed as part of the Drosophila genome mapping project [Sidén-Kiamos, I., Saunders, R.D.C., Spanos, L., Majerus, T., Trenear, J., Savakis, C., Louis, C., Glover, D.M., Ashburner, M. & Kafatos, F.C. (1990) Nucleic Acids Res. 18, 6261-6270]. The 5' non-coding region of the isolated gene hybridised to cytological position 13C1-2. By combining reverse transcription and the polymerase chain reaction, the gene was shown to be expressed at a very low level. The PP1 13C gene encodes a protein of 302 amino acids with a predicted molecular mass of 34.5 kDa. It shows 85-94% amino acid identity to the other three protein phosphatase 1 catalytic subunits (PP1 87B, PP1 96A and PP1 9C) described previously, being most closely related to the isoform PP1 87B, which is involved in the control of chromosome separation at cell division and the regulation of chromosome condensation at interphase.

Amino Acid Sequence↗

The antioxidant LY231617 reduces global ischemic neuronal injury in rats.

BACKGROUND AND PURPOSE: In the rat four-vessel occlusion model with 30 minutes of ischemia most agents have failed to be of benefit when given after ischemia. Because postischemia administration is more clinically relevant, we evaluated the antioxidant LY231617 (2,6-bis(1,1-dimethylethyl)-4-[[(1-ethyl)amino]methyl]phenol hydrochloride]) when administered after 30 minutes of four-vessel occlusion. METHODS: Male Wistar rats were subjected to 30 minutes of four-vessel occlusion. LY231617 was either given orally 30 minutes before ischemia or intravenously beginning at 30 minutes after the onset of ischemia. Hippocampal CA1 layer and striatal damage were rated on a scale of 0-3 (0, no damage; 3, > 90% cell loss). We also evaluated the ability of LY231617 to prevent iron-dependent lipid peroxidation and to prevent hydrogen peroxide-induced neuronal death of hippocampal neurons in primary culture by exposing cultures to a 50-microM concentration of hydrogen peroxide for 15 minutes in the presence of LY231617. RESULTS: Oral administration of LY231617 reduced both striatal and hippocampal CA1 damage by > 75% (p < 0.0001). In two separate experiments in which LY231617 was given intravenously beginning 30 minutes after occlusion, hippocampal and striatal damage were reduced by approximately 50% (p < 0.03) in the first experiment and by approximately 41% (p < 0.02) in the second experiment. Addition of 5 microM of LY231617 to primary hippocampal neuronal cultures antagonized the lethal effect of hydrogen peroxide (p < 0.05). Iron-dependent lipid peroxidation was also inhibited in a dose-related fashion. CONCLUSIONS: The significant reduction of ischemia-induced or hydrogen peroxide-induced neuronal damage and inhibition of lipid peroxidation by LY231617 observed in this study suggest that reactive oxygen intermediates play an important role in the events leading to neuronal death after global ischemia/reperfusion.

Animals↗

Mutations in the Drosophila melanogaster gene three rows permit aspects of mitosis to continue in the absence of chromatid segregation.

We have cloned the three rows (thr) gene, by a combination of chromosome microdissection and P element tagging. We describe phenotypes of embryos homozygous for mutations at the thr locus. Maternal mRNA and protein appear to be sufficient to allow 14 rounds of mitosis in embryos homozygous for thr mutations. However, a small percentage of cells in syncytial blastoderm stage thr embryos sink into the interior of the embryo as if they have failed to divide properly. Following cellularisation all cells complete mitosis 14 normally. All cells become delayed at mitosis 15 with their chromosomes remaining aligned on the spindle in a metaphase-like configuration, even though both cyclins A and B have both been degraded. As cyclin B degradation occurs at the metaphase-anaphase transition, subsequent to the microtubule integrity checkpoint, the delay induced by mutations at the thr locus defines a later point in mitotic progression. Chromosomes in the cells of thr embryos do not undertake anaphase separation, but remain at the metaphase plate. Subsequently they decondense. A subset of nuclei go on to replicate their DNA but there is no further mitotic division.

Amino Acid Sequence↗

Dodeca satellite: a conserved G+C-rich satellite from the centromeric heterochromatin of Drosophila melanogaster.

To identify sequences from the centromeric region, we have constructed a Drosophila melanogaster yeast artificial chromosome (YAC) library and screened it with purified DNA from the minichromosome Dp(1;f)1187 derived from the X chromosome. We describe the structure of one clone isolated in this way. This YAC is structurally unstable and contains tandemly repeated G+C-rich 11-mer and 12-mer units, which we call dodeca satellite. Most of this satellite is located near the centromere of an autosome. Cross-hybridizing sequences are found in the genomes of organisms as distant as Arabidopsis thaliana and Homo sapiens.

Animals↗