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Biomedical subjects

R Hull

Publications and source records attributed to R Hull.

At least 181 records · Page 10Linked to original sources
PubMed1983

Isolation and characterization of faithful and altered clones of the genomes of cauliflower mosaic virus isolates Cabb B-JI, CM4-184, and Bari I.

Full-length genomes of cauliflower mosaic virus (CaMV) isolates Cabb B-JI, CM4-184, and Bari I have been cloned in the SalGI site of plasmid pAT 153. The cloned DNAs were characterized by restriction mapping and infectivity assays. All the sites present in the virion DNAs were found in the cloned DNAs. Comparison of restriction maps with those of DNA from two other isolates which have been recently completely sequenced revealed a close relationship among the different isolates. Some of the clones appear to be faithful copies of the viral genomes and these viral inserts are infectious when inoculated into turnip plants. Various clones with deletions in the CaMV DNA have been isolated and characterized. Some of them may correspond to deletions naturally occurring in a subpopulation of the virus whereas others occurred during cloning. None of the deleted fragments are infectious when inoculated into plants. Strikingly, all the deletions overlap one or two of the specific single-stranded breaks characteristic of caulimoviruses, suggesting that sequences surrounding the breaks are not dispensable.

Chromosome Mapping↗
PubMed1983

The application of spot hybridization to the detection of DNA and RNA viruses in plant tissues.

A solid-phase nucleic acid hybridization technique for the detection of DNA and RNA viruses in plant tissues is described. The method involves spotting crude samples onto nitrocellulose and using 12P-labelled DNA hybridization probes. The limit of sensitivity is 5-20 pg virus/spot or approximately 5 micrograms/g leaf tissue. The method is quantitative for DNA viruses in crude homogenates, but not for RNA viruses. The amount of cauliflower mosaic virus in infected leaves and protoplasts was estimated. The amplitude of spot hybridization to screening plant material from glasshouses and field is discussed.

DNA, Viral↗
PubMed1983

A simple procedure for the extraction of double-stranded RNA from virus-infected plants.

A simple procedure for the isolation of double-stranded (ds) RNA from virus-infected plants is described. The method is based on grinding plant tissue in 4% p-aminosalicylic acid and recovery of ds RNA by phenol extraction and precipitation with 30% ethanol. The presence of both negative and positive virus RNA strands in RNA fractionated in agarose gels was verified by Northern blot hybridization with polynucleotide kinase labelled genomic RNA or complementary DNA (cDNA) probes. The procedure enabled detection of three major ds RNA species (MWs 4.2, 1.05 and 0.48 X 10(6)) and at least 4 minor bands with estimated MWs of 3.5, 2.5, 2.2 and 2.0 X 10(6) in Nicotiana tabacum plants systemically infected with tobacco mosaic virus (TMV). Cucumber mosaic virus (CMV)-infected Pachystachys coccinea plants contained 2 minor bands of MWs 0.49 and 0.35 X 10(6) in addition to the previously described 4 major ds RNAs and ds CARNA 5 (MW 0.22 X 10(6)). The patterns of ds RNA are useful for diagnosing natural infections of CMV and TMV in N. glauca plants and of citrus tristeza virus in Citrus spp.

Mosaic Viruses↗
PubMed1983

Contribution of adhesion to bacterial persistence in the mouse urinary tract.

The affinity of uropathogenic Escherichia coli to kidneys and bladders of experimentally infected mice was shown to be determined in part by the adhesive properties of the infecting bacteria. Mice were infected with various pairwise combinations of two homogeneic sets of bacteria: (i) mutants derived from a human pyelonephritis E. coli isolate which were selected to express either or both adhesins specific for globoseries glycolipid receptors or for "mannosides"; and (ii) transformants of a normal fecal isolate which harbored recombinant plasmids encoding the genes for one or the other adhesin or which harbored only the vector plasmid. The relative efficiency of survival of the strains to be compared was evaluated in each animal by plating on selective media of samples of homogenized kidneys and bladders taken 24 h after intravesical inoculation. The presence of adhesins specific for globoseries glycolipid receptors, which mediate the in vitro mannose-resistant attachment to human and mouse uroepithelial cells, enhanced bacterial recovery from both kidneys and bladders of infected animals. The addition to the infecting strain of adhesins binding mannoside residues further improved bacterial recovery from the bladder, but not from the kidney. The mutants and transformants with adhesins binding only mannosides were recovered in higher numbers from the bladder than those expressing adhesins specific for the globoseries glycolipids only. There was apparent selection in vivo decreasing expression of mannoside binding adhesins in the kidneys, but not in the bladders, of animals infected with the mutant expressing both types of adhesins. Regardless of adhesive properties, the mutants of the pyelonephritis isolate were recovered in significantly higher numbers than the fecal isolate with adhesins encoded on recombinant plasmids. We conclude that the adhesive properties in part determine the localization and retention of bacteria in the mouse urinary tract. However, the addition of adhesins to a commensal E. coli strain was not sufficient to confer colonization capacity comparable to that of a pyelonephritis strain.

Adhesins, Escherichia coli↗
PubMed1983

Genetics of digalactoside-binding adhesin from a uropathogenic Escherichia coli strain.

The uropathogenic strain Escherichia coli J96 mediates mannose-resistant hemagglutination owing to production of a digalactoside-binding adhesin. A cosmid clone from this strain has been isolated that, when harbored in E. coli K-12, expressed Pap pili and this adhesin (R. Hull et al., Infect. Immun. 33:933-938, 1981). By transposon mutagenesis and by the construction of a number of hybrid plasmid derivatives, we have demonstrated that about 8.5 kilobases of DNA is required to generate a mannose-resistant hemagglutination-positive phenotype in E. coli K-12 strain P678-54. The structural gene for the Pap pili monomer, papA, has been identified and mapped close to the promotor-proximal end of the Pap operon. Although strain P678-54 that harbored a Tn5 insertion within papA showed a mannose-resistant hemagglutination-positive phenotype, it was negative in a competitive enzyme-linked immunosorbent assay with anti-Pap pilus serum. This could mean that a Pap adhesin is encoded by a region on the Pap operon that is distinct from papA.

Adhesins, Escherichia coli↗
PubMed1983

Molecular cloning and physical characterization of a chromosomal hemolysin from Escherichia coli.

A chromosomal hemolysin determinant was isolated in the form of recombinant plasmid pSF4000 from a human urinary tract isolate. Escherichia coli J96. A restriction endonuclease fragment map of pSF4000 was constructed. The ampicilin resistance transposon Tn/ was used as a site-specific mutagen in conjunction with in vitro derived deletions to localize the hemolysin determinant to a 7.0-kilobase region of pSF4000. Seventeen hemolytic E. coli isolates from human urinary tract infections were found to share similar DNA sequences with J96 hemolysin sequences, using a hemolysin-specific restriction endonuclease fragment as a hybridization probe. In all 17 hemolytic E. coli strains the hemolysin gene sequences were localized to the bacterial chromosome. The hemolysin-specific gene sequences can be found only in hemolytic strains by colony blot hybridization, suggesting that in the evolutionary sense the hemolysin-specific genetic sequences have only recently been introduced into the E. coli chromosome. Evidence is also presented that in the regions neighboring the J96 chromosomal hemolysin there are additional unique DNA sequences not commonly found in other E. coli isolates.

Base Sequence↗
PubMed1983

Target cell specificity of wild-type E. coli and mutants and clones with genetically defined adhesins.

The species, individual and tissue specificity of bacterial binding reactions was studied using wild-type E. coli strains from diarrhoea or urinary tract infection, and derivatives with genetically manipulated adhesins. E. coli J96 and GR12 were isolated from the urine of patients with acute pyelonephritis; E. coli strains expressing the CFAI and II antigens from the stools of patients with diarrhoea. E. coli J96, GR12 and CFAI induced mannose-resistant agglutination of human erythrocytes; E. coli J96 and GR12 in addition carried mannose-sensitive adhesins. Mutants of GR12 with either or both of these adhesins were obtained through chemical mutagenesis. Cloning of 6-8 mdal fragments of chromosomal DNA from J96 into E. coli K12 resulted in expression of pili and binding properties in the previously non-piliated and non-binding strain. Bacterial binding was registered to target cells from different human tissues; small intestinal brush borders, uroepithelial and buccal cells and erythrocytes and was compared between species using rabbit intestinal brush borders, mouse bladder cells and guinea pig erythrocytes. Individual variation was illustrated by agglutination of human P1 erythrocytes and those of blood group p lacking the globoseries glycolipid receptors. Specific recognition of globoseries glycolipid receptors was defined as capacity to agglutinate guinea pig erythrocytes after but not before coating with globotetraosylceramide. Binding specific for mannose-containing receptors was diagnosed by mannose-reversible agglutination of guinea pig erythrocytes. The binding pattern of the wild-type strains was related to the site of infection, i.e. the CFAI and II strains bound to small intestinal brush borders and the pyelonephritogenic E. coli to uroepithelial cells, but not vice versa. The mutants and clones retained the binding properties of the parent/donor both in degree and specificity of binding. Strains with adhesins specific for globoseries glycolipids attached to human uroepithelia, mouse uroepithelial and buccal cells. Within the group of strains with mannose-sensitive adhesins heterogeneity was observed. Strains sharing ability to agglutinate guinea pig erythrocytes to a mannose-reversible manner bound or did not bind to human buccal cells, to human uroepithelial cells and agglutinated or did not agglutinate human erythrocytes. The results demonstrate the usefulness of genetic technology in the study of bacterial binding reactions. The role of pili as adhesins is discussed.

Adhesins, Escherichia coli↗
PubMed1982

Different intensities of oral anticoagulant therapy in the treatment of proximal-vein thrombosis.

We have previously reported that long-term therapy with warfarin is effective for preventing recurrent venous thromboembolism in patients with proximal-vein thrombosis but that there is an appreciable risk of hemorrhage. To determine whether that risk could be reduced without a loss of effectiveness, we randomly allocated 96 patients with proximal-vein thrombosis to a group receiving less intense anticoagulant therapy, with a mean prothrombin time of 26.9 seconds using the Manchester comparative reagent (corresponding Simplastin time, 15 seconds), or a group given more intense therapy, with a mean Simplastin time of 19.4 seconds (corresponding prothrombin time 41 seconds with the Manchester comparative reagent) (P less than 0.001). Two of 47 patients (4 per cent) in the less intensely treated group had hemorrhagic complications, as compared with 11 of 49 patients (22 per cent) in the more intensely anticoagulated group (P = 0.015 by the two-tailed test). This difference was due to minor bleeding episodes. The frequency of recurrent venous thromboembolism was low in both groups (2 per cent). Our findings indicate that less intense anticoagulant therapy is associated with a low frequency of recurrent venous thromboembolism (2 per cent) and a reduced risk of hemorrhage.

Administration, Oral↗
PubMed1982

Inhibition of experimental ascending urinary tract infection by an epithelial cell-surface receptor analogue.

It has been shown that the establishment of urinary tract infection by Escherichia coli is dependent on attachment of the bacteria to epithelial cells. The attachment involves specific epithelial cell receptors, which have been characterized as glycolipids. Reversible binding to cell-surface mannosides may also be important. This suggests an approach to the treatment of infections--that of blocking bacterial attachment with cell membrane receptor analogues. Using E. coli mutants lacking one or other of the two binding specificities (glycolipid and mannose), we show here that glycolipid analogues can block in vitro adhesion and in vivo urinary tract infection.

Animals↗
PubMed1982

Adjusted subcutaneous heparin versus warfarin sodium in the long-term treatment of venous thrombosis.

Previously, we compared fixed low doses of heparin with adjusted doses of warfarin for the long-term treatment of venous thrombosis; in that study low-dose heparin was ineffective in preventing recurrence in patients with proximal-vein thrombosis. We have now completed a randomized trial comparing adjusted doses of heparin and of warfarin for prevention of recurrent venous thromboembolism in patients with proximal-vein thrombosis. One hundred six consecutive patients with acute proximal-vein thrombosis confirmed by venography were treated with intravenous heparin and then randomized to secondary prophylaxis. Two of 53 patients receiving heparin, as compared with one of 53 receiving warfarin, had new episodes of objectively documented venous thromboembolism. Nine patients taking warfarin had bleeding complications (which were major in three patients), as compared with one patient taking heparin (P = 0.008). Our data indicate that adjusted-dose subcutaneous heparin therapy provides an effective alternative to warfarin sodium and is associated with a lower risk of bleeding.

Clinical Trials as Topic↗
PubMed1982

The diagnostic value of the fibrinogen/fibrin fragment E antigen assay in clinically suspected deep vein thrombosis.

We have evaluated the fibrinogen/fibrin fragment E antigen assay as a diagnostic test in patients with clinically suspected venous thrombosis by comparing the results of this assay with venography in 272 patients. The result of the fragment E antigen assay was elevated in 79 of 80 patients with positive venograms for recent venous thrombosis (sensitivity 99%) and within the normal range in 161 of 192 patients with normal venograms (specificity 84%). The fragment E assay was also evaluated in 130 medical and surgical controls without evidence of venous thrombosis by leg scanning and the test was found to be relatively nonspecific. However, in the patient group under study, a correct clinical diagnosis of no thrombosis, based on a normal fragment E result, was made in 161 of 162 cases (negative predictive value of 99%). Therefore, a normal test result effectively excludes a diagnosis of venous thrombosis in clinically symptomatic patients. The assay, as currently performed, is technically demanding and takes 24 hr to complete. Therefore, it will have to be simplified before it can be applied to clinical practice.

Adolescent↗