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Biomedical subjects

J Gray

Publications and source records attributed to J Gray.

At least 19 recordsLinked to original sources
PubMed1992

X-ray laser microscopy of rat sperm nuclei.

The development of high brightness and short pulse width (< 200 picoseconds) x-ray lasers now offers biologists the possibility of high-resolution imaging of specimens in an aqueous environment without the blurring effects associated with natural motions and chemical erosion. As a step toward developing the capabilities of this type of x-ray microscopy, a tantalum x-ray laser at 44.83 angstrom wavelength was used together with an x-ray zone plate lens to image both unlabeled and selectively gold-labeled dried rat sperm nuclei. The observed images show approximately 500 angstrom features, illustrate the importance of x-ray microscopy in determining chemical composition, and provide information about the uniformity of sperm chromatin organization and the extent of sperm chromatin hydration.

Animals↗
PubMed1992

Deletion of chromosome 17p loci in breast cancer cells detected by fluorescence in situ hybridization.

Allelic loss of tumor suppressor genes on chromosome 17p has been implicated in the progression of breast cancer. This is in principle detectable by fluorescence in situ hybridization if the loss occurs by deletion. In order to determine if detectable deletions occur in primary breast cancer, we used dual-color hybridization with chromosome 17 pericentromeric and region-specific DNA probes to study 19 primary breast cancers. The copy numbers of 17 centromere and 17p13.1 sequences were compared with the loss of heterozygosity (LOH) for probe YNZ22 at 17p13.3 detected by restriction fragment length polymorphism. Nine of 11 cases showing LOH also showed the major population of nuclei with a deletion. The remaining two tumors with LOH were trisomic for both the centromere and 17p13.1 cosmid. In contrast, seven of eight tumors without LOH had no deletions by fluorescence in situ hybridization. These data suggest that the dominant mechanism of allelic loss at 17p in breast cancer is a physical deletion and that analysis of deletions by fluorescence in situ hybridization is a rapid and sensitive approach to studying chromosomal aberrations.

Alleles↗
PubMed1992

Use of recombinant biotinylated aequorin in microtiter and membrane-based assays: purification of recombinant apoaequorin from Escherichia coli.

Aequorin is a calcium-dependent bioluminescent protein isolated from the hydromedusan Aequorea victoria. The gene for aequorin has been cloned and overexpressed in Escherichia coli [Prasher et al. (1985) Biochem. Biophys. Res. Commun. 126, 1259; Prasher et al. (1987) Biochemistry 26, 1326]. Higher levels of expression have recently been obtained by subcloning aequorin cDNA into the pRC23 plasmid vector such that its expression is under control of the lambda PL promoter [Cormier et al. (1989) Photochem. Photobiol. 49, 509]. Purification of recombinant apoaequorin from E. coli containing this new recombinant plasmid (pAEQ1.3) was accomplished by a two-step procedure involving gel filtration and anion-exchange chromatography on Sephadex G-100 and DEAE-Sepharose, respectively. Typically, 400-500 mg of recombinant protein was obtained from 100 L of fermentation culture. The purified recombinant apoaequorin could be converted to aequorin in high yield upon incubation with synthetic coelenterate luciferin, dissolved oxygen, and a thiol reagent with a photon yield similar to the native photoprotein. Detection of recombinant aequorin in the Dynatech ML1000 Microplate luminometer was linear between 10(-18) and 10(-12) mol, and little loss of specific activity was observed when the protein was derivatized with biotin. The biotinylated derivative was stable when frozen, lyophilized, or stored at 4 degrees C. The feasibility of using biotinylated aequorin as a nonradioactive tag was established by its application in a variety of solid-phase assay formats using the high-affinity streptavidin/biotin interaction. A microtiter-based bioluminescent immunoassay (BLIA) using biotinylated aequorin and the ML1000 luminometer was developed for the detection of subnanogram amounts of a glycosphingolipid (Forsmann antigen). In addition, nanogram to subnanogram quantities of protein antigens and DNA, immobilized on Western and Southern blots, respectively, were detected on instant and X-ray films using biotinylated aequorin.

Aequorin↗
PubMed1992

Molecular biology of fruit ripening and its manipulation with antisense genes.

Considerable progress in tomato molecular biology has been made over the past five years. At least 19 different mRNAs which increase in amount during tomato fruit ripening have been cloned and genes for enzymes involved in cell wall degradation (polygalacturonase and pectinesterase) and ethylene synthesis (ACC synthase) have been identified by conventional procedures. Transgenic plants have been used to identify regions of DNA flanking fruit-specific, ripening-related and ethylene-regulated genes and trans-acting factors which bind to these promoters have also been identified. Antisense genes expressed in transgenic plants have proved to be highly effective for inhibiting the specific expression of ripening-related genes. These experiments have changed our understanding of how softening occurs in tomato fruit. Antisense techniques have also been used to identify genes encoding enzymes for carotenoid biosynthesis (phytoene synthase) and ethylene biosynthesis (the ethylene-forming enzyme). The altered characteristics of fruit transformed with specific antisense genes, such as retarded ripening and resistance to splitting, may prove to be of value to fruit growers, processors and ultimately the consumer.

Antisense Elements (Genetics)↗
PubMed1992

Construction and characterization of a yeast artificial chromosome library containing 1.5 equivalents of human chromosome 21.

A library of yeast artificial chromosomes (YACs) was constructed from a human/hamster somatic cell hybrid containing human chromosome 21 (q11-qter). Cells were embedded in agarose, and the DNA was partially digested with EcoRI, released into solution by agarase treatment of the agarose plugs, ligated into pYAC4, and transferred into yeast. Double screening of the yeast transformants with human and hamster genomic DNA allowed the selection of clones hybridizing only with human DNA. The library consists of 321 clones, amounting to 1.5 equivalents (61 Mb) of chromosome 21. The mean YAC size calculated from 178 clones is 190 +/- 100 kb. Screening of the library with eight sequence-tagged sites gave six positives. Among 21 YACs tested by in situ hybridization, 17 mapped to chromosome 21.

Animals↗
PubMed1992

Rapid translocation frequency analysis in humans decades after exposure to ionizing radiation.

This paper presents an analysis of the utility of fluorescence in situ hybridization (FISH) with whole-chromosome probes for measurement of the genomic frequency of translocations found in the peripheral blood of individuals exposed to ionizing radiation. First, we derive the equation: Fp = 2.05fp(1-fp)FG, relating the translocation frequency, Fp, measured using FISH to the genomic translocation frequency, FG, where fp is the fraction of the genome covered by the composite probe. We demonstrate the validity of this equation by showing that: (a) translocation detection efficiency predicted by the equation is consistent with experimental data as fp is changed; (b) translocation frequency dose-response curves measured in vitro using FISH agree well with dicentric frequency dose-response curves measured in vitro using conventional cytogenetic procedures; and (c) the genomic translocation frequencies estimated from FISH measurements for 20 Hiroshima A-bomb survivors and four workers exposed to ionizing radiation during the Y-12 criticality accident are approximately the same as the translocation frequencies measured using G-banding. We also show that translocation frequency dose response curves estimated using FISH are similar for Hiroshima A-bomb survivors and for first division lymphocytes irradiated in vitro. We conclude with a discussion of the potential utility of translocation frequency analysis for assessment of the level of acute radiation exposure independent of the time between analysis and exposure.

Accidents↗
PubMed1992

Classical conditioning and cognitive determinants of subjective craving for opiates: an investigation of their relative contributions.

Subjective craving for drugs is frequently reported by detoxified addicts. Explanatory models emphasizing classical conditioning (CC) and cognitive mechanisms have been proposed, and the mechanisms postulated by the two approaches are discussed here. There are three major variants of the CC model, arguing that craving is associated with conditioned changes which are respectively drug-agonistic, opponent-process, or withdrawal reactions. The cognitive model highlights the role of 'outcome expectancies' as determinants of both appetitive and avoidant motivations. The present investigation has identified correlates of craving predicted by the above models, and tested them empirically. Detoxified opiate addicts participated in in a Craving Test requiring them to rate their craving, physical state, and emotions during exposures to both neutral and drug-related material. Other measures, including personality and outcome expectancies for drug use, were also taken, and interrelationships among the variables were explored statistically. Whilst some of the predictions of the CC formulations were borne out, the cognitive model received more consistent support. The theoretical and treatment implications of these data are considered.

Adult↗
PubMed1992

Mutation of the miaA gene of Agrobacterium tumefaciens results in reduced vir gene expression.

vir regulon expression in Agrobacterium tumefaciens involves both chromosome- and Ti-plasmid-encoded gene products. We have isolated and characterized a new chromosomal gene that when mutated results in a 2- to 10-fold reduction in the induced expression of vir genes by acetosyringone. This reduced expression occurs in AB minimal medium (pH 5.5) containing either sucrose or glucose and containing phosphate at high or low concentrations. The locus was cloned and used to complement A. tumefaciens strains harboring Tn5 insertions in the gene. Sequence analysis of this locus revealed an open reading frame with strong homology to the miaA locus of Escherichia coli and the mod5 locus of Saccharomyces cerevisiae. These genes encode tRNA: isopentenyltransferase enzymes responsible for the specific modification of the A-37 residue in UNN codon tRNA species. The function of the homologous gene in A. tumefaciens was proven by genetic complementation of E. coli miaA mutant strains. tRNA undermodification in A. tumefaciens miaA mutant strains may reduce vir gene expression by causing a reduced translation efficiency. A slight reduction in the virulence of these mutant Agrobacterium strains on red potato plants, but not on tobacco, tomato, kalanchoe, or sunflower plants, was observed.

Acetophenones↗
PubMed1992

Specificity of the proteus antibody response in rheumatoid arthritis.

Antibodies to proteus were determined by indirect immunofluorescence in 146 serum samples from patients with rheumatoid arthritis (RA). An autoantibody screen was performed in the same samples and in 52 of these antibody titres to the viruses influenza A, adenovirus, rubella, and parvovirus were determined. There was no significant correlation between proteus antibodies and any of the other antibodies tested. Dividing the samples into those from patients with active (C reactive protein > 10 mg/l) and inactive RA showed that the only antibodies to be significantly increased in active RA were the proteus antibodies. These observations suggest that the proteus antibody response in RA is specific.

Acute Disease↗
PubMed1992

P blood group phenotype, proteus antibody titres, and rheumatoid arthritis.

The interrelationships between P blood group phenotype, proteus antibodies and rheumatoid arthritis (RA) were investigated in 140 patients with RA and 114 of their siblings who did not have RA. In the group with RA P2 subjects had significantly higher titres of proteus antibodies than P1 patients. This was not observed in the group without RA, or for antibodies to Escherichia coli. Although C reactive protein was the best predictor of proteus antibodies in the group with RA, the P blood group had an independent and significant influence. These observations suggest a testable model in which asymptomatic carriage of proteus in the urinary tract may lead to antibody production, which in turn may be important in the pathogenesis of RA.

Acute Disease↗
PubMed1992

Changing patterns of malaria in south-east Scotland: implications for practitioner awareness and prophylactic advice.

The medical records of all 229 patients with malaria admitted to the Edinburgh City Hospital between 1969 and 1988 were studied retrospectively. A total of 137 were from Africa, 44 from the Indian subcontinent, 19 from the Far East, 18 from New Guinea, 5 from the Middle East and 3 from South America. The number of yearly admissions rose markedly after 1983, mainly due to an increase in Plasmodium falciparum cases. Ninety-four cases (15 with severe parasitaemia) mainly from Kenya and Nigeria were due to P. falciparum infection and 99 to P. vivax. There were no deaths. A seasonal distribution of onset of fever in patients with P. vivax infections originating from the Indian subcontinent showed that most patients presented during the summer. Prophylaxis had generally been irregular or non-existent but many compliant patients may have been receiving an inadequate dose of chloroquine on a mg/kg body weight basis. General practitioners are likely to see at least one case of malaria every 4 years. They are encouraged to seek advice from a specialist unit whenever necessary whether before or after their patient travels abroad. Travellers, in particular to Kenya and Nigeria, and Asian immigrants to the UK returning on holiday to their country of origin should be strongly advised to take regular prophylaxis including on return to the UK.

Adolescent↗
PubMed1992

Factors that determine the in vivo dose-response relationship for stable chromosome aberrations in A-bomb survivors.

An overview is given of the dose-response relationship for stable chromosome aberrations (i.e., translocations and inversions) in the peripheral blood lymphocytes of A-bomb survivors in Hiroshima. Special emphasis is placed on (i) the overdispersion of survivor cases with either unexpectedly high or low aberration frequencies relative to the estimated DS86 kerma values assigned to individual survivors, termed "cytogenetic outliers", and (ii) the correlation of chromosome aberration frequencies with other biological endpoints, such as acute radiation symptoms (severe epilation). A new molecular biological technique, known as fluorescence in situ hybridization (FISH) with composite, whole-chromosome probes to paint differentially the target chromosomes, has facilitated rapid, efficient, and extensive scoring of translocation-type chromosome aberrations in which the target chromosomes are involved. Using this methodology, the observed findings on translocation frequencies in A-bomb survivors have shown that the frequency of stable chromosome aberrations, which have persisted for years without change in frequency in irradiated persons, is indeed useful as an indicator for biological dosimetry.

Chromosome Aberrations↗
PubMed1992

Yersinia-related arthritis in the United Kingdom. A report of 12 cases and review of the literature.

Reactive arthritis following infection with Yersinia is endemic in Scandinavian countries; the prevalence is low in the UK, however. We have reviewed the literature pertaining to Yersinia-related reactive arthritis in the UK and describe 12 patients who presented over a 3-year period with an asymmetrical seronegative polyarthropathy and serological evidence of recent Yersinia infection. Five patients recalled having a diarrhoeal illness prior to the onset of the arthropathy. None had a prior history of psoriasis, inflammatory bowel disease or ankylosing spondylitis. A history of urethral discharge was elicited from one patient. Extra-articular manifestations were seen in three patients (iritis in two, erythema nodosum in another). Four patients developed chronic joint disease after periods of 4, 6, 8, and 18 months, respectively. The prevalence of Yersinia-related arthritis in the UK may be higher than previously thought.

Adolescent↗
PubMed1992

A randomized trial of nimodipine in severe head injury: HIT I. British/Finnish Co-operative Head Injury Trial Group.

We studied the efficacy of nimodipine in severely head-injured patients in a randomized study. Of 176 patients who received nimodipine, 2 mg/h iv for 2 day, 53% had a favorable outcome (moderate or good recovery). Of 175 control patients, 49% had a favorable outcome. This difference was not statistically significant but does not exclude the possibility that a study of a larger number of patients could show a clinically useful benefit.

Adult↗
PubMed1991

An NF kappa B-like factor is essential but not sufficient for cytokine induction of endothelial leukocyte adhesion molecule 1 (ELAM-1) gene transcription.

The endothelial leukocyte adhesion molecule 1 (ELAM-1) is transiently expressed specifically on the surface of cytokine-induced endothelial cells. We demonstrate that the transient expression of the protein is paralleled by an increase and decrease in transcription of the ELAM-1 gene. To identify the cis-acting transcription control regions within the ELAM-1 gene that are responsible for this cytokine-induced expression, we isolated and analyzed an ELAM-1 genomic clone containing sequences upstream of the transcription start site. We constructed a series of ELAM-1 deletion mutants linked to a reporter gene and analyzed their expression in both endothelial and non-endothelial cells. Results show that a fragment of 233 bp upstream of the transcription start site is sufficient to confer cytokine inducibility upon the reporter gene in both endothelial and non-endothelial cells. Further analysis defined two elements within this region that are involved in the cytokine inducibility of the ELAM-1 gene. One element lies within the -233 to -117 region, the other element represents an NF kappa B consensus binding site between nucleotides -94 to -85. Gel shift analysis reveals increased binding of an NF kappa B-like factor to this consensus sequence in extracts prepared from IL-1-induced endothelial cells. The results suggest that cytokine induction of ELAM-1 gene transcription is imparted by a combination of positive factors, one being an NF kappa B-like transcription factor, interacting with cis-acting elements within the enhancer/promoter of the gene.

Base Sequence↗