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L Coates

Publications and source records attributed to L Coates.

29 records · Page 2Linked to original sources

In vitro infection of woodchuck hepatocytes with woodchuck hepatitis virus and ground squirrel hepatitis virus.

Primary cultures of woodchuck hepatocytes were demonstrated to be susceptible to in vitro infection by both woodchuck hepatitis virus and ground squirrel hepatitis virus, as evidenced by the appearance of DNA species characteristic of hepadnavirus replication. Initiation of infection by woodchuck hepatitis virus was blocked by the presence of suramin, polybrene, or dideoxycytidine. Viral CCC DNA, the putative template for viral RNA transcription, was detected at 2 days postinfection. Accumulation of intracellular intermediates in virion DNA synthesis was negligible until 7-10 days postinfection, but these DNA intermediates then increased dramatically in amount over the next few weeks. Results were obtained which suggested that the prolonged accumulation of intermediates in virion DNA synthesis was an intrinsic property of the infection of individual cells, and not the result of a slow spread of virus through the cultures.

Animals↗

Molecular cloning of hepatitis delta virus RNA from an infected woodchuck liver: sequence, structure, and applications.

cDNA prepared from the single-stranded circular RNA genome of hepatitis delta virus was cloned in lambda gt11 by using RNA from the liver of an infected woodchuck. From the sequence of overlapping clones, we assembled the full sequence of 1,679 nucleotides. The sequence indicated an exceptional ability for intramolecular base pairing, yielding a rod structure with at least 70% of the bases paired and a predicted free energy of -805 kcal (-3,368 kJ)/mol. Three of the lambda clones contained sequences that were not only expressed as fusion proteins with beta-galactosidase but were recognized by human hepatitis delta virus-specific antibody. These clones were sequenced so as to establish the reading frame of the delta antigen on the antigenomic strand. The fusion protein produced by one clone was purified by immunoaffinity chromatography and then was used to raise rabbit antibodies specific for the delta antigen.

Animals↗

Suramin inhibits in vitro infection by duck hepatitis B virus, Rous sarcoma virus, and hepatitis delta virus.

Suramin blocked in vitro infection by duck hepatitis B virus, a hepadnavirus, and Rous sarcoma virus, a retrovirus. Although suramin was able to inhibit the virus-encoded reverse transcriptase activities of these two viruses, this inhibition did not appear to account for the anti-viral effect of the drug. In particular, suramin was unable to block synthesis within cells of full-length viral DNAs when added subsequent to infection. The results are consistent with the hypothesis that suramin acted by blocking virus uptake or uncoating. As further support of this hypothesis, we found that suramin also blocked infection by hepatitis delta virus, an RNA virus that is not known to employ reverse transcriptase during the initiation of infection.

Animals↗

Humoral immune responsiveness in duck hepatitis B virus-infected ducks.

Immunofluorescence assays with fixed tissue sections were used to characterize antibody reactivity in sera obtained from duck hepatitis B virus-infected ducks. Under conditions of experimental infection, antibody to core antigen but not to surface antigen was detectable. A majority of the ducks infected at 8 days after hatching and a minority of those infected at 1 day after hatching showed a transient anti-core antigen humoral response; this response was stronger in the antibody-positive ducks infected on day 8 than in those infected on day 1. Antiviral antibody was not detected in the sera of ducks congenitally infected with duck hepatitis B virus. Several of the infected ducks, but none of the uninfected ducks, exhibited autoantibody reactivity for alpha-islet-cell-associated antigen.

Animals↗

Replication of human hepatitis delta virus in primary cultures of woodchuck hepatocytes.

We obtained two lines of evidence that monolayer cultures of primary woodchuck hepatocytes support replication of the genome of human hepatitis delta virus (HDV). (i) From a Northern (RNA blot) analysis of the HDV-related RNA in infected cultures, both genomic and antigenomic 1.7-kilobase RNA species were detected at 11 days after infection. The ratio of genomic RNA to antigenomic RNA was 2:1 to 10:1, comparable to that previously reported in studies of experimentally infected chimpanzees and woodchucks. (ii) Replication in culture was also demonstrated by in situ hybridization with a strand-specific probe. Such studies showed that only a small fraction of the cultured cells supported replication and that in such cells the relative and absolute levels of the HDV RNAs were comparable to those in liver cells infected in vivo. Furthermore, as with the in vivo studies, the HDV RNAs were predominantly localized to the nucleus. In summary, we demonstrated that cultured cells supported both the early events of HDV adsorption and penetration and the intermediate events of genome replication.

Animals↗

Experimental transmission of duck hepatitis B virus.

Susceptibility to experimental infection with duck hepatitis B virus (DHBV) was explored, with the objective of defining procedures that were both rapid and reproducible. For the purpose of these experiments, a small flock of DHBV-free breeders was established as a source of susceptible eggs and ducklings, since ca. 10% of the ducks (all ages) from commercial flocks were DHBV infected. Intravenous inoculation of DHBV into 15-day duck embryos from the DHBV-free flock produced a persistent infection, with a high-titer viremia, in at least 80% of the injected animals. The tissue tropism of DHBV in these experimentally infected animals was similar to that associated with natural, congenital infections from viremic ducks to their progeny. Virus antigen was found not only in hepatocytes and bile duct epithelium of liver, but also in cells associated with exocrine and endocrine pancreas, and in proximal convoluted tubular epithelium of kidney. Infection of embryonic liver was rapid, as evidenced by active synthesis of DHBV-DNA by reverse-transcription of RNA by 24 hr postinjection. During this latter analysis, formation of supercoiled viral DNA appeared to precede the reverse-transcription phase of viral DNA synthesis, suggesting that this species may be important in initiation of infection.

Animals↗

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Child Development↗

Care of the ventilated patient.

Practice profiles are reflective pieces written by nurses in practice and based on continuing professional development articles. This week Laura Coates discusses mechanical ventilation. Article No. 503. Henderson N (1999) Mechnical ventilation.

Humans↗