Search PubMed⌕ Search

Biomedical subjects

R W Morgan

Publications and source records attributed to R W Morgan.

At least 37 records · Page 2Linked to original sources
PubMed1994

Enhanced expression of the Marek's disease virus-specific phosphoproteins after stable transfection of MSB-1 cells with the Marek's disease virus homologue of ICP4.

Phosphoprotein pp38, coded for by the BamHI-H fragment of the Marek's disease herpesvirus (MDV) genome is expressed in tumor cells and tumor cell lines. pp38 is associated with two other phosphoproteins, pp41 and pp24, and can be detected in a small percentage of tumor cells by indirect immunofluorescence assays (IIFA). The importance of MDV ICP4 for the regulation of pp38 expression was examined in the following MSB-1-derived cell lines stably transfected with the selection plasmid pNL1 [MDCC-CU221 (CU221)], pNL1 and the BamHI-A fragment of MDV DNA containing ICP4 (CU224), MDV ICP4 inserted in antisense direction in the eukaryotic expression vector pXT1 (CU222), or ICP4 in sense direction in pXT1 (CU223) or cotransfected with pNL1 and EcoRI-linearized BamHI-A MDV DNA (CU225, -237, -243, -244). IIFA analysis showed that CU223 had a markedly increased expression of pp38, while CU224 had a slightly increased expression. No changes were noted in CU221 or CU222, while expression of pp38 was decreased in CU225, -237, -243, and -244. Radioimmunoprecipitation assays demonstrated that the expression of all three phosphoproteins was enhanced in CU223. Steady-state transcriptional analysis showed that CU223 had increased levels of pp38-specific (1.9 and 3.3 kb) and ICP4-specific (10.0 kb) transcripts.

Animals↗
PubMed1994

Characterization of a Marek's disease virus mutant containing a lacZ insertion in the US6 (gD) homologue gene.

We report the construction of a Marek's disease virus (MDV) mutant containing the lacZ gene of Escherichia coli inserted into a homologue of the US6 (glycoprotein D, gD) gene of herpes simplex virus. The mutant was constructed using the high-passage GAatt85 MDV strain as the parent virus, since that strain grows readily in chicken embryo fibroblasts using culture conditions conducive to mutant virus construction. The lacZ insertion site was positioned one third of the way into the US6 (gD) open reading frame. Insertion of the lacZ gene disrupted a major 6.2 kb transcript that initiated approximately 2.5 kb upstream of the gD homologue gene in the vicinity of the US3 homologue and sorf4 genes, and extended into the US7 (gI) homologue gene. The mutant virus (US6lac) and the parent virus had similar growth kinetics in cell culture at 37 degrees C and 41 degrees C. Furthermore, the US6lac mutant could be reisolated from the spleens and peripheral blood of infected chickens with a frequency comparable to that of the parent virus. Our results indicate that the gene encoding the gD homologue is nonessential for growth in cell culture or for infection of chickens following intra-abdominal inoculation with an attenuated serotype-1 MDV.

Animals↗
PubMed1994

Identification of latency-associated transcripts that map antisense to the ICP4 homolog gene of Marek's disease virus.

Two small RNAs (0.9 and 0.75 kb), named Marek's disease virus (MDV) small RNAs (MSRs) and a 10-kb RNA, all of which map antisense to the MDV ICP4 homolog gene, have been readily detected in MDCC-MSB1 MDV-transformed T-lymphoblastoid cells. These RNAs were not detectable in reticuloendotheliosis virus-transformed T cells. When MDV was reactivated by treatment of lymphoblastoid cells with 25 micrograms of iododeoxyuridine per ml, the relative levels of the transcripts decreased. These RNAs were not detected by Northern (RNA) hybridization in productively infected chicken embryo fibroblasts 48 h postinfection; however, they were apparent 140 h postinfection. By using Northern hybridization, RNase protection assays, and primer extension analysis, the MSRs were determined to map antisense to the predicted translational start site of the ICP4 homolog gene. The conclusion most consistent with the data is that the two MSRs are overlapping, spliced RNAs. Both small RNAs contain a latency promoter binding factor consensus recognition sequence located toward their 5' ends as well as two potential ICP4 recognition consensus sequences, one in each orientation. The region contains a number of small open reading frames on each side and within the MSRs. Although the exact endpoints are unknown, the large 10-kb species spans the entire ICP4 homolog region. We believe that this group of RNAs, which map antisense to the ICP4 homolog gene, are latency-associated transcripts of MDV.

Animals↗
PubMed1994

Characterization of Marek's disease virus insertion and deletion mutants that lack US1 (ICP22 homolog), US10, and/or US2 and neighboring short-component open reading frames.

We report the characterization of Marek's disease virus (MDV) strains having mutations in various genes that map to the unique short (US) region of the viral genome. A deletion mutant (GA delta 4.8lac) lacks 4.8 kbp of US region DNA, the deleted segment having been replaced by the lacZ gene of Escherichia coli. This deletion results in the loss of the MDV-encoded US1, US10, and US2 homologs of herpes simplex virus type 1, as well as three putative MDV-specific genes, Sorf1, Sorf2, and Sorf3. Two mutants containing lacZ insertions in the US1 and US10 genes have been constructed, and we have previously reported a US2lac insertion mutant (J. L. Cantello, A. S. Anderson, A. Francesconi, and R. W. Morgan, J. Virol. 65:1584-1588, 1991). The isolation of these mutants indicates that the relevant genes are not required for growth of MDV in chicken embryo fibroblasts. The mutants had early growth kinetics indistinguishable from those of their parent viruses; however, 5 to 7 days after being plated, the US1 insertion mutant (US1lac) and the GA delta 4.8lac deletion mutant showed a 5- to 10-fold decrease in virus growth. This decrease in virus accumulation correlated with a 30 to 50% decrease in plaquing efficiency when these viruses were plated onto established versus fresh chicken embryo fibroblast monolayers compared with a 10 to 15% decrease seen for the parent viruses and for the US10lac or US2lac insertion mutants. Finally, GA delta 4.8lac could be reisolated from chickens, indicating that the deleted genes are not required for the infection of chickens following intra-abdominal inoculation of an attenuated serotype 1 MDV.

Animals↗
PubMed1993

Update on artificial sweeteners and bladder cancer.

Review of the most recent epidemiologic studies indicates there is no detectable association between artificial sweetener consumption and bladder cancer. The summary relative risk from a meta-analysis of all case-control studies approaches unity (RR .97). This analysis, in combination with new evidence for a unique mechanism of saccharin carcinogenicity in the male rat, leads to the conclusion that saccharin is not related to bladder cancer in humans.

Animals↗
PubMed1993

Avian herpesvirus as a live viral vector for the expression of heterologous antigens.

Control of Marek's disease in the poultry industry has been successfully achieved for several decades by large-scale vaccination of day-old chickens with live herpesvirus of turkeys (HVT) strains. Several features of this virus including lack of pathogenicity and long-term immune protection due to a persistent viraemic infection made us decide to use HVT as a live viral vector for the expression of foreign antigens. Potential sites for the integration of foreign DNA in the unique short region of the HVT genome were identified by the insertion of a beta-galactosidase expression cassette. Vaccination trials with recombinant virus strains indicated that the marker gene was expressed and stably maintained during animal passage. Based on an insertion site mapping in one of the open reading frames of the unique short region, a general recombination vector was designed for the integration of foreign genes into HVT. Recombinant virus-directed expression of individual antigens from Newcastle disease virus was driven by a strong promoter element derived from the lung terminal repeat sequence of Rous sarcoma virus.

Amino Acid Sequence↗
PubMed1992

Complete nucleotide sequence of the Marek's disease virus ICP4 gene.

The Marek's disease virus (MDV) gene encoding a homologue to the ICP4 protein of herpes simplex virus has been mapped to BamHl fragment A based on the physical map of the MDV genome (Fukuchi et al., 1984). The gene lies completely within the inverted repeat flanking the unique short region of the genome. The complete nucleotide sequence of the MDV ICP4 gene has been determined. The coding region is 4245 nucleotides long and has an overall G+C content of 52%. The MDV ICP4 protein is predicted to have a structure similar to that of ICP4-like proteins of other herpesviruses in that it has five distinct regions, the second and fourth of which are highly conserved. In addition, the protein contains the characteristic run of serine residues located toward its amino terminus. The MDV ICP4 gene is expressed in MDV-infected chicken embryo fibroblasts.

Amino Acid Sequence↗
PubMed1992

Characterization of reticuloendotheliosis virus-transformed avian T-lymphoblastoid cell lines infected with Marek's disease virus.

The expression of Marek's disease virus (MDV) transcripts and protein products was investigated in reticuloendotheliosis virus-transformed avian T-lymphoblastoid cell line RECC-CU91, which was superinfected with MDV. The presence of MDV in the superinfected cell line, renamed RECC-CU210, was demonstrated by Southern hybridization with 32P-labeled BamHI-H and -B fragments of the BamHI MDV DNA library. Examination of RECC-CU210 for the expression of MDV-specific RNA transcripts encoded by the internal repeat long (IRL), internal repeat short (IRS), and unique short (US) regions of the MDV genome revealed two small transcripts of 0.6 and 0.7 kb. These transcripts were mapped to the IRL and IRS regions, respectively. In contrast, RECC-CU211, which was developed through transfection of CU210 with the BamHI-A fragment of MDV, expressed an additional nine transcripts from the IRL, IRS, and US regions. CU211 but not CU210 also expressed a complex of polypeptides of 40, 38, and 24 kDa, identified by monoclonal antibodies as MDV-specific phosphoproteins. The 38-kDa phosphoprotein is likely to be pp38, an early viral protein that maps within the IRL region of the MDV genome. These findings suggest that genes located within the transfected BamHI-A fragment transactivated a number of genes located in the IRL region of the MDV genome.

Animals↗
PubMed1991

The solvent effects of calcium hydroxide irrigating solution on bovine pulp tissue.

The solvent effects of calcium hydroxide irrigating solution (used alone and in combination with sodium hypochlorite) on bovine pulp tissue were studied. Forty pieces of pulp tissue weighing 90 mg each were treated with calcium hydroxide solution alone, calcium hydroxide and sodium hypochlorite alternated, sodium hypochlorite alone, and saline alone. Each piece of tissue was treated for 32 min. Desiccated pretreatment and posttreatment weights were compared. There was no significant difference between the dissolution capability of calcium hydroxide solution used alone and of saline. No significant difference was noted between calcium hydroxide solution and sodium hypochlorite used alternately, and sodium hypochlorite used alone. However, both of these groups were significantly more effective at dissolving tissue than calcium hydroxide solution alone or saline. Calcium hydroxide solution was an ineffective solvent of pulpal tissue. If tissue dissolution is desired during root canal therapy, the use of calcium hydroxide solution as the sole irrigant is no more effective than saline.

Analysis of Variance↗
PubMed1991

Isolation of a Marek's disease virus (MDV) recombinant containing the lacZ gene of Escherichia coli stably inserted within the MDV US2 gene.

We have isolated a stable, recombinant Marek's disease virus (MDV) containing the lacZ gene of Escherichia coli inserted into the unique short region of the genome. The nucleotide sequence of the insertion site indicates that it lies within a sequence homologous to the US2 gene of herpes simplex virus. Stable insertion of the lacZ gene into the MDV US2 gene indicates that the site is nonessential for MDV growth in cell culture.

Amino Acid Sequence↗
PubMed1991

Whodunit? Liability for mesothelioma cases.

It is possible to apportion blame for mesothelioma incidence to previous exposure in relation to an exponent of the time since that exposure. This article demonstrates the application of a formula to calculate the probability that a specific exposure caused an individual mesothelioma case.

Asbestosis↗
PubMed1989

An alkyl hydroperoxide reductase from Salmonella typhimurium involved in the defense of DNA against oxidative damage. Purification and properties.

A peroxide reductase (peroxidase) which converts lipid hydroperoxides and other alkyl hydroperoxides to the corresponding alcohols, using either NADH or NADPH as the reducing agent, has been identified in both Salmonella typhimurium and Escherichia coli. This enzyme is shown to play a role in protecting against alkyl hydroperoxide mutagenesis. To our knowledge this work represents the first description of an NAD(P)H peroxidase in enteric bacteria and the first reported bacterial peroxidase to exhibit high activity toward alkyl hydroperoxides. A high performance liquid chromatography-based assay for the alkyl hydroperoxide reductase has been developed by monitoring the reduction of cumene hydroperoxide, a model alkyl hydroperoxide. By using this assay, the enzyme has been purified from a S. typhimurium regulatory mutant, oxyR1, which overexpresses a number of proteins involved in defenses against oxidative damage, and which contains 20-fold more of the alkyl hydroperoxide reductase than the wild-type strain. The purified activity requires the presence of two separable components having subunit molecular weights of 22,000 and 57,000. The 57-kDa protein contains a bound FAD cofactor and can use either NADH or NADPH as an electron donor for the direct reduction of redox dyes, or of alkyl hydroperoxides when combined with the 22-kDa protein. This enzyme may thus serve as a prokaryotic equivalent to the glutathione reductase/glutathione peroxidase system in eukaryotes.

Benzene Derivatives↗
PubMed1989

An epidemiologic investigation of birth outcomes in relation to dibromochloropropane contamination in drinking water in Fresno County, California, USA.

This report describes an epidemiologic investigation of the relationship between dibromochloropropane (DBCP), contamination in drinking water and birth outcomes between 1978 and 1982 in Fresno County, California, USA. Census tracts in the county were categorized according to DBCP level in drinking water. A direct comparison between exposure categories was done by the use of the Mantel-Haenszel chi-square procedure to compute relative ratios or risks for low birth weight and birth defect, adjusting for age, race, percent Hispanic, and parity. No correlation between these birth outcomes and DBCP contamination was found. In addition, we did not find any difference in sex ratio in relation to DBCP contamination.

Adolescent↗
PubMed1989

An alkyl hydroperoxide reductase induced by oxidative stress in Salmonella typhimurium and Escherichia coli: genetic characterization and cloning of ahp.

The ahp genes encoding the two proteins (F52a and C22) that make up an alkyl hydroperoxide reductase were mapped and cloned from Salmonella typhimurium and Escherichia coli. Two classes of oxidant-resistant ahp mutants which overexpress the two proteins were isolated. ahp-1 was isolated in a wild-type background and is dependent on oxyR, a positive regulator of defenses against oxidative stress. ahp-2 was isolated in an oxyR deletion background and is oxyR independent. Transposons linked to ahp-1 and ahp-2 or inserted in ahp mapped the genes to 13 min on the S. typhimurium chromosome, 59% linked to ent. Deletions of ahp obtained in both S. typhimurium and E. coli resulted in hypersensitivity to killing by cumene hydroperoxide (an alkyl hydroperoxide) and elimination of the proteins F52a and C22 from two-dimensional gels and immunoblots. ahp clones isolated from both S. typhimurium and E. coli complemented the cumene hydroperoxide sensitivity of the ahp deletion strains and restored expression of the F52a and C22 proteins. A cis-acting element required for oxyR-dependent, rpoH-independent heat shock induction of the F52a protein was present at the S. typhimurium but not the E. coli ahp locus.

Blotting, Western↗
PubMed1989

Ecological analyses and case-control studies of gastric cancer and leukaemia in relation to DBCP in drinking water in Fresno County, California.

Through ecological analyses and case-control studies, the possible relation of gastric cancer and leukaemia to dibromochloropropane (DBCP) contamination of drinking water in Fresno County, California, has been examined. The ecological analyses examined the correlation between gastric cancer and leukaemia (including the lymphatic varieties), mortality rates, and DBCP concentrations in drinking water by census tract in Fresno County, 1960-83. No correlation was found between gastric cancer or leukaemia and DBCP. The gastric cancer case-control study consisted of 263 deaths from gastric cancer in the county, 1975 to mid-1984, and 1044 controls, using information on residential history and occupation of both cases and controls. Analyses were based on residence at death, as well as one and ten years before death. The case-control study did not find any relation between gastric cancer and DBCP in drinking water. Hispanics in the county were found to experience a relative risk of gastric cancer of 2.77, compared with non-Hispanics. A similar case-control study consisting of 259 cases of leukaemia and 1161 controls found no relation between all leukaemia or lymphatic leukaemia and DBCP in drinking water. Farm workers, however, do appear to have an increased risk of leukaemia.

Agricultural Workers' Diseases↗