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Biomedical subjects

J A Lewis

Publications and source records attributed to J A Lewis.

At least 91 records · Page 5Linked to original sources
PubMed1989

Interferon-induced expression of different genes is mediated by distinct regulatory pathways.

Interferons (IFNs) regulate the expression of multiple genes and the regulatory sequences associated with several of these genes have been identified. How cell-surface IFN receptors communicate with such regulatory elements is as yet unknown. We have characterized the IFN responses of a mutant murine cell line (Ltk-aprt-) which is unable to mount an antiviral response when treated with IFN-beta. In contrast to its inability to activate antiviral pathways in these cells, IFN-beta inhibits their growth to the same level as observed in their parental L-929 line which is fully responsive to IFN, suggesting that distinct pathways account for antiviral and cytostatic responses. In agreement with this, Northern blot and nuclear run-on analyses show that the induction of transcription of three distinct genes (2,5(A) synthetase, BS-I-150, and BS-II-4) is blocked in Ltk-aprt- cells whereas another gene (I-8) is activated normally. Our results show that the defective responses observed in this cell line are not due to a nonfunctional IFN-beta cell-surface receptor and suggest that multiple pathways exist between the receptor and upstream gene regulatory elements.

2',5'-Oligoadenylate Synthetase↗
PubMed1989

Beta and gamma interferons act synergistically to produce an antiviral state in cells resistant to both interferons individually.

We showed previously that the mouse fibroblastoid cell line Ltk-aprt- is resistant to the antiviral effects of beta interferon. This lack of response reflects a partial sensitivity to the interferon that is accompanied by a failure to activate expression of several interferon-regulated genes, although certain other genes respond in a normal manner. We show here that Ltk-aprt- cells were also unable to establish an antiviral state and to activate expression of 2,5-oligo(A) synthetase when treated with gamma interferon. Strikingly, however, treatment with a combination of beta interferon and gamma interferon provided complete protection against viral replication. Although the cells were completely insensitive to up to 250 U of the interferons per ml added singly, essentially complete protection from viral cytopathic effects was achieved when as little as 10 U of each of the interferons per ml were combined. Expression of 2,5-oligo(A) synthetase was also sensitive to this synergistic effect. Activation of an antiviral state could also be achieved by sequential treatment, first with gamma interferon and then with beta interferon. Partial protection against viral replication could be achieved by pretreatment with gamma interferon for as little as 1 h before incubation with beta interferon and could be blocked by the addition of specific antibodies or by cycloheximide, indicating that gamma interferon induces the synthesis of a protein which can act synergistically with a signal produced by the beta-interferon receptor. We suggest that Ltk-aprt- cells suffer from defects in one or more components of the gene activation pathways for both type I and type II interferons. Nonetheless, gamma interferon is able to activate the expression of a gene encoding a protein required for signal transduction. This protein acts synergistically with a transient signal produced in response to beta interferon, thereby activating the expression of a further group of genes.

2',5'-Oligoadenylate Synthetase↗
PubMed1989

Beta-blockers in portal hypertension. An overview.

This paper describes a meta-analysis of data from the 5 primary and 15 secondary prevention studies in patients with portal hypertension, in which beta-blockers were compared with inactive control. These studies involved 1525 patients. Overall, the number of patients who suffered bleeding events was 38% lower in the groups treated with beta-blockers (p less than 0.001). The percentage reductions in bleeding-related deaths and in total mortality were 35% and 18%, respectively (p = 0.007 and p = 0.024). More detailed analysis supports the conclusion that propranolol, the beta-blocker predominantly used in these studies, is effective in both the primary and secondary prevention of bleeding events and mortality due to portal hypertension.

Adrenergic beta-Antagonists↗
PubMed1988

Induction of an antiviral state by interferon in the absence of elevated levels of 2,5-oligo(A) synthetase and eIF-2 kinase.

A series of clones has been derived from an interferon-resistant murine cell line, Ltk- aprt-, and their antiviral properties have been characterized. In the parental Ltk- aprt- line interferon is unable to establish antiviral properties or to increase the levels of 2,5-oligo(A) synthetase, the 2,5-oligo(A)-activated endonuclease F, 2',5'-phosphodiesterase, or eIF-2 kinase. However, interferon did prevent replication of vesicular stomatitis, Mengo virus, and reovirus in some of the derivative cell lines. The effect of interferon on the levels of the enzymes of the 2,5-oligo(A) and eIF-2 kinase pathways did not correlate directly with the antiviral properties of these cell clones. Greatly increased levels of 2,5-oligo(A) synthetase occurred in one clone without activation of an antiviral state. Another clone exhibited antiviral activity without detectably increased 2,5-oligo(A) synthetase activity. Changes in the levels of endonuclease F and 2',5'-phosphodiesterase were slight in all the clones examined. Neither 2,5-oligo(A) synthetase nor eIF-2 kinase levels were altered by interferon in another clone and yet an antiviral state was established and prevented replication of vesicular stomatitis, Mengo virus, and reovirus. The results show that mechanisms other than the 2,5-oligo(A) and eIF-2 kinase pathways are likely to contribute to the antiviral effects of interferon.

2',5'-Oligoadenylate Synthetase↗
PubMed1988

A microtiter cell-culture assay for the determination of anti-human immunodeficiency virus neutralizing antibody activity.

A microtiter cell-culture assay is described for measuring neutralizing antibody activity directed against the human immunodeficiency virus (HIV). The assay relies upon inhibition of HIV-mediated cell killing of infected MT-4 lymphoid cells. The assay exhibits comparable sensitivity to two other methods used for such measurements, is relatively rapid, may be adapted for screening large numbers of samples and involves minimal handling of infectious virus.

Antibodies, Viral↗
PubMed1987

Activation of metallothionein expression is potentiated by DNA sequences present in the herpes simplex virus thymidine kinase gene.

A mouse cell line (Ltk-aprt-) which is resistant to the anti-viral effects of interferon also has a reduced ability to synthesize metallothionein on exposure to cadmium. Like the ability to respond to interferon, cadmium-induced metallothionein synthesis is restored to wild-type levels in clones obtained by introducing a thymidine kinase gene into Ltk-aprt-cells. Transfection of other genes does not have such an effect. Since metallothionein expression is also activated by interferon the results suggest that the regulation of several genes which are responsive to interferon can be modulated by specific sequences present in the Herpes virus thymidine kinase gene.

Animals↗
PubMed1987

Migraine trials: crossover or parallel group?

This paper evaluates the relative merits of the crossover design and the parallel group design for the evaluation of migraine therapy, focussing on prophylaxis. Three issues are highlighted which lead to a preference for the parallel group trial: changes in the treatment effect with time, the possibility of carry-over effects, and the handling of withdrawals and protocol deviations. These have to be weighed against the greater power of the crossover design. It is shown that the difference in power is relatively small when baseline or run-in assessments are available. It is therefore concluded that the parallel group trial is the design of choice for prophylaxis, and that it has a place in the evaluation of acute therapy.

Clinical Trials as Topic↗
PubMed1987

Parasites, diseases and health status of sympatric populations of sambar deer and white-tailed deer in Florida.

From December 1983 to December 1984 a study on parasites, diseases and health status was conducted on sympatric populations of sambar deer (Cervus unicolor) and white-tailed deer (Odocoileus virginianus) from St. Vincent Island, Franklin County, Florida. Ten sambar and six white-tailed deer were examined. White-tailed deer had antibodies to epizootic hemorrhagic disease virus and bluetongue virus. Serologic tests for antibodies to the etiologic agents of bovine virus diarrhea, infectious bovine rhinotracheitis, vesicular stomatitis, parainfluenza 3, brucellosis, and leptospirosis were negative in both species of deer. White-tailed deer harbored 19 species of parasites; all were typical of the parasite fauna of this species in coastal regions of the southeastern United States. Sambar deer harbored 13 species of parasites, which apparently were derived largely from white-tailed deer. The only exception was Dermacentor variabilis which occurs frequently on wild swine on the island. The general health status of sambar deer appeared to be better than that of white-tailed deer. This was hypothesized to result from the sambar deer's utilization of food resources unavailable or unacceptable to white-tailed deer and to the absence and/or lower frequency of certain pathogens in sambar deer.

Animals↗
PubMed1987

Cholinergic receptor mutants of the nematode Caenorhabditis elegans.

Potential acetylcholine receptor (AChR) mutants of the nematode are selectable by resistance to the neurotoxic drug levamisole, a probable cholinergic agonist. To determine which mutants may have achieved resistance through loss of levamisole receptor function, we have assayed mutant extracts for specific 3H-meta-aminolevamisole binding activity in the presence and absence of mecamylamine. We find that mutants in 3 of the 7 genes associated with extreme levamisole resistance are obviously deficient in saturable specific 3H-meta-aminolevamisole binding activity. Mutants of the 4 other genes have abnormal binding activities that fail to undergo the apparent allosteric activation of saturable specific 3H-meta-aminolevamisole binding activity caused by mecamylamine. Thus, all 7 genes appear to be required to produce a fully functional levamisole receptor. Mutants of several other genes associated only with partial resistance to levamisole have at least grossly normal receptor binding activities.

Animals↗
PubMed1987

The levamisole receptor, a cholinergic receptor of the nematode Caenorhabditis elegans.

We describe a glass fiber filter binding assay for the levamisole receptor, a putative acetylcholine receptor of the nematode Caenorhabditis elegans, and we show that receptor detected in vitro binds both levamisole derivatives and cholinergic agonists with the pharmacological specificity expected of the physiologically functional nematode receptor. The receptor is detected by the binding of tritiated meta-aminolevamisole ([3H]MAL, 27 Ci/mmol). In extracts of the wild-type nematode, there is a saturable, high affinity binding activity for [3H]MAL (Kd approximately 5-10 nM). Well fed wild-type worms contain as much as 3 fmol of high affinity binding activity per mg of extract protein (0.14 pmol/g of wet weight of worms) and dauer larvae, a special juvenile stage, contain as much as 15 fmol of activity per mg of protein. Specific binding activity per mg of protein is highest in larval stages and decreases severalfold in the adult worm. The rates of formation and dissociation of the [3H]MAL-receptor complex are relatively slow (dissociation half-life, 17 min), in agreement with physiological studies of levamisole on Ascaris muscle strips. Levamisole derivatives and cholinergic agonists have the same relative potencies in inhibiting [3H]MAL binding as they do in causing nematode muscle contraction. Vertebrate cholinergic antagonists do not inhibit [3H]MAL binding, but several antagonists (mecamylamine, alpha-bungarotoxin, and cobra venom) potentiate the binding of [3H]MAL and can be used to demonstrate more clearly the presence of a second, lower affinity binding activity whose ligand-binding affinity is also potentiated by these agents. Both high and low affinity wild-type binding components are missing in the extremely levamisole-resistant mutant unc-74(x19).

Age Factors↗