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M Lovett

Publications and source records attributed to M Lovett.

At least 91 records · Page 5Linked to original sources
PubMed1990

Effects of the lethal yellow (Ay) mutation in mouse aggregation chimeras.

The Ay allele is a recessive lethal mutation at the mouse agouti locus, which results in embryonic death around the time of implantation. In the heterozygous state, Ay produces several dominant pleiotropic effects, including an increase in weight gain and body length, a susceptibility to hepatic, pulmonary and mammary tumors, and a suppression of the agouti phenotype, which results in a yellow coat color. To investigate the cellular action of Ay with regard to its effects upon embryonic viability and adult-onset obesity, we generated a series of aggregation chimeras using embryos that differ in their agouti locus genotype. Embryos derived from Ay/a x Ay/a matings were aggregated with those derived from A/A x A/A matings, and genotypic identification of the resultant chimeras was accomplished using a molecular probe at the Emv-15 locus that distinguishes among the three different alleles, Ay, A, and a. Among 50 chimeras, 25 analyzed as liveborns and 25 as 9.5 day embryos, 29 were a/a in equilibrium A/A and 21 were Ay/a in equilibrium A/A. The absence of Ay/Ay in equilibrium A/A chimeras demonstrates that Ay/Ay cells cannot be rescued in a chimeric environment, and the relative deficiency of Ay/a in equilibrium A/A chimeras suggests that, under certain conditions, Ay heterozygosity may partially affect cell viability or proliferation. In the 25 liveborn chimeras, Ay/a in equilibrium A/A animals became obese as adults and a/a in equilibrium A/A animals did not. There was no correlation between genotypic proportions and rate of weight gain, which shows that, with regard to its effects on weight gain, Ay heterozygosity is cell non-autonomous.

Animals↗
PubMed1988

Intravenous acyclovir therapy of first episodes of genital herpes: a multicenter double-blind, placebo-controlled trial.

PURPOSE: A collaborative multicenter double-blind, placebo-controlled trial of intravenous acyclovir treatment of first-episode genital herpes was performed in order to substantiate previous findings on the efficacy and safety of this drug, to evaluate the influence of parenteral therapy on recurrence frequency, and to obtain further data on the natural history of genital herpes. PATIENTS AND METHODS: Eighty-two patients with first episodes of genital herpes simplex virus (HSV) infection were randomly assigned in a double-blind fashion to treatment with intravenous acyclovir (5 mg/kg every eight hours) or placebo for five days. Before therapy, all lesions in the genital/perineal area and in extragenital sites were cultured. New lesions appearing in both areas after the onset of therapy were cultured separately. Lesions in all groups were cultured until completely healed. Sera were collected from all patients on entry to the study and on Day 21 to determine presence or absence of antibodies to HSV-1 and HSV-2. Time to healing, time to crusting, time to cessation of viral shedding, and appearance of new lesions during therapy were compared for each treatment group. RESULTS: Patients receiving acyclovir experienced a significant reduction in the median duration of pain (4.3 versus 4.8 days, p = 0.019), viral shedding (1.9 versus 8.4 days, p less than 0.001), and time to healing (8.4 versus 11.5 days, p = 0.02) compared with placebo recipients. These differences were largely attributable to the effect of therapy in the subset of patients with primary disease in whom acyclovir reduced the median duration of pain from 10.6 days to 4.2 days, the median duration of viral shedding from 17.1 days to 1.9 days, and the median time to healing from 14.2 days to 8.3 days. The rate of subsequent recurrence of genital herpes was not altered by acyclovir treatment: 24 of 32 acyclovir recipients (75 percent) experienced one or more recurrences during a mean follow-up of 14 months compared with 19 of 27 placebo recipients (70 percent). Among patients experiencing recurrences, the mean number of recurrences per month among acyclovir recipients was 0.25 compared with 0.19 for patients given placebo. CONCLUSION: This multicenter trial confirms the efficacy of intravenous acyclovir in the management of first-episode genital herpes, especially in patients with primary infection. However, therapy did not alter the frequency of recurrences.

Acyclovir↗
PubMed1988

Genetics and polymorphism of the mouse prion gene complex: control of scrapie incubation time.

The mouse prion protein (PrP) gene (Prn-p), which encodes the only macromolecule that has been identified in scrapie prions, is tightly linked or identical to a gene (Prn-i) that controls the duration of the scrapie incubation period in mice. Constellations of restriction fragment length polymorphisms distinguish haplotypes a to f of Prn-p. The Prn-pb allele encodes a PrP that differs in sequence from those encoded by the other haplotypes and, in inbred mouse strains, correlates with long scrapie incubation time (Westaway et al., Cell 51: 651-662, 1987). In segregating crosses of mice, we identified rare individuals with a divergent scrapie incubation time phenotype and Prn-p genotype, but progeny testing to demonstrate meiotic recombination was not possible because scrapie is a lethal disease. Crosses involving the a, d, and e haplotypes demonstrated that genes unlinked to Prn-p could modulate scrapie incubation time and that there were only two alleles of Prn-i among the mouse strains tested. All inbred strains of mice that had the Prnb haplotype were probably direct descendants of the I/LnJ progenitors. We established the linkage relationship between the prion gene complex (Prn) and other chromosome 2 genes; the gene order, proximal to distal, is B2m-II-1a-Prn-Itp-A. Recombination suppression in the B2m-Prn-p interval occurred during the crosses involved in transferring the I/LnJ Prnb complex into a C57BL/6J background. Transmission ratio distortion by Prna/Prnb heterozygous males was also observed in the same crosses. These phenomena, together with the founder effect, would favor apparent linkage disequilibrium between Prn-p and Prn-i. Therefore, transmission genetics may underestimate the number of genes in Prn.

Animals↗
PubMed1987

The mouse homolog of the human amyloid beta protein (AD-AP) gene is located on the distal end of mouse chromosome 16: further extension of the homology between human chromosome 21 and mouse chromosome 16.

The human amyloid beta protein is the major constituent of the brain amyloid plaques found in Alzheimer disease. The gene that encodes this protein is located on chromosome 21, and individuals with Down syndrome (trisomy 21) also exhibit an early onset form of Alzheimer disease. We have used the cloned human amyloid beta protein gene and a panel of somatic cell hybrids to map the location of the mouse homolog of this gene. We report here that the mouse gene is located on chromosome 16 within the region 16C3----ter, in common with three other genes which map within the Down syndrome region of human chromosome 21.

Amyloid↗
PubMed1987

Transgenic mice with increased Cu/Zn-superoxide dismutase activity: animal model of dosage effects in Down syndrome.

Down syndrome, the phenotypic expression of human trisomy 21, is presumed to result from a 1.5-fold increase in the expression of the genes on human chromosome 21. As an approach to the development of an animal model for Down syndrome, several strains of transgenic mice that carry the human Cu/Zn-superoxide dismutase gene have been prepared. These animals express the transgene in a manner similar to that of humans, with 0.9- and 0.7-kilobase transcripts in a 1:4 ratio, and synthesize the human enzyme in an active form capable of forming human-mouse enzyme heterodimers. Cu/Zn-superoxide superoxide dismutase activity is increased from 1.6- to 6.0-fold in the brains of four transgenic strains and to an equal or lesser extent in several other tissues. These animals provide a unique system for studying the consequences of increased dosage of the Cu/Zn-superoxide dismutase gene in Down syndrome and the role of this enzyme in a variety of other pathological processes.

Alzheimer Disease↗
PubMed1987

The lethal yellow allele-associated provirus results in the production of chimeric viral-host RNAs.

The lethal yellow (Ay) mutation is the most dominant allele at the agouti locus of the mouse. Mice heterozygous for this allele have a yellow coat color, are genetically obese with an increased susceptibility to cancer, and have other metabolic derangements. Mice homozygous for Ay die early in embryogenesis, possibly because of a trophectoderm defect. The Ay mutation is distinguished from the many alleles described at the agouti locus in being associated with an endogenous provirus, designated endogenous ecotropic murine leukemia viral locus 15 (Emv-15). To obtain DNA sequences from regions close to or within the agouti locus, we have isolated the Emv-15 provirus and have found that the DNA sequences adjacent to the provirus are part of a mouse gene that is expressed in the same transcriptional orientation as the proviral genes. These mouse DNA sequences recognize two distinct size classes of RNA in various adult wild-type tissues including skin. In Ay heterozygotes both types of transcript hybridize to the proviral long terminal repeat, and, in heterozygous spleens, the shorter transcript is present at an enhanced steady-state level. These results suggest that the Ay-associated provirus (Emv-15) is distinguished from the endogenous C-type proviruses described to date in having altered the expression of a flanking host gene by promoter insertion, resulting in the production of chimeric viral-host fusion transcripts.

Alleles↗
PubMed1987

Molecular markers for the agouti coat color locus of the mouse.

The agouti (a) coat color locus of the mouse acts within the microenvironment of the hair follicle to control the relative amount and distribution of yellow and black pigment in the coat hairs. Over 18 different mutations with complex dominance relationships have been described at this locus. The lethal yellow (Ay) mutation is the top dominant of this series and is uniquely associated with an endogenous provirus, Emv-15, in three highly inbred strains. However, we report here that it is unlikely that the provirus itself causes the Ay-associated alteration in coat color, since one strain of mice (YBR-Ay/a) lacks the provirus but still retains a yellow coat color. Using single-copy mouse DNA sequences from the regions flanking Emv-15 we have detected three patterns of restriction fragment length polymorphisms (RFLPs) within this region that can be used as molecular markers for different agouti locus alleles: a wild-type agouti (A) pattern, a pattern which generally cosegregates with the nonagouti (a) mutation, and a pattern which is specific to Emv-15. We have used these RFLPs and a panel of 28 recombinant inbred mouse strains to determine the genetic linkage of these sequences with the agouti locus and have found complete concordance between the two (95% confidence limit of 0.00 to 3.79 centimorgans). We have also physically mapped these sequences by in situ hybridization to band H1 of chromosome 2, thus directly confirming previous assignments of the location of the agouti locus.

Alleles↗
PubMed1986

Isolation of transcribed DNA sequences from chromosome 21 using mouse fetal cDNA.

A technique for isolating DNA sequences that are likely to be transcriptionally active in both humans and mice has been developed. This method is based on the screening of a human genomic DNA library with single stranded cDNA prepared from late gestation mouse fetal RNA. Using a human chromosome 21 DNA library, we have isolated and characterized two clones which are entirely composed of single copy human DNA sequences and are 2.7 and 6.7 kilobases in length. The 2.7 kilobase clone is homologous to a transcript found predominantly in nonpolyadenylated human fibroblast RNA. It also shows homology to mouse genomic DNA and recognises several polyadenylated RNAs in mouse testis. This cloned sequence has been mapped by in situ hybridization to the distal third of chromosome 21, the region involved in the causation of Down syndrome.

Animals↗
PubMed1986

The mouse IFN-alpha (Ifa) locus: correlation of physical and linkage maps by in situ hybridization.

The physical location of the mouse IFN-alpha locus (Ifa) on chromosome 4 was defined by in situ hybridization of a cloned mouse IFN-alpha probe to metaphase spreads in which one chromosome 4 was present as part of a single metacentric chromosome, all other chromosomes being acrocentric. (This approach greatly facilitates analysis and can be used even when it is difficult to obtain good banding). Using unbanded chromosomes, the grains were localized over the chromosome 4 part of the metacentric, in a region 0.61 +/- 0.07 (SD) of the distance from the centromere to the telomere. In Giemsa-banded spreads, the majority of the grains were in the region 4C3----C6. Consideration of these results and of the known linkage maps for mouse and man indicates that the Galt - Aco-1 - Ifa syntenic group spans a distance of approximately 14 cM and suggests that the same group on human 9p will also occupy a similarly sized region, with GALT proximal and IFL distal to the centromere.

Animals↗
PubMed1985

Treatment of first episode genital HSV with oral acyclovir: long term follow-up of recurrences. A preliminary report.

Systemic acyclovir (ACV) treatment has been shown to have significant effects on shortening the clinical course of first episode genital HSV infections, decreasing the quantity of HSV antibody in convalescent phase serum, without affecting the incidence of recurrences in 6 months of follow-up. In order to assess long term recurrence patterns, we prospectively studied subjects enrolled in a randomized double blind trial of oral ACV for first episode genital HSV. Sixty-three of sixty-eight subjects were followed monthly for recurrences for a mean of 36 months with 90% of subjects completing two years. There was no difference in the incidence of recurrence (90%) in (29) placebo vs (37) ACV treated HSV-2 infected subjects. Recurrences rates were similar between ACV and placebo treated subjects with nonprimary HSV-2 infection followed 2 years. The majority of these subjects were found to have HSV-2 antibody in their acute sera suggesting prior asymptomatic acquisition of HSV-2 infection and therefore established ganglionic latency at the time of first clinical disease. In subjects with true primary HSV-2 infection, however, mean recurrence rates were significantly lower in ACV treated subjects after 6 months, 0.87 ACV vs 3 placebo per 6 month period (p less than 0.01). The percentage of subjects experiencing recurrences after 1 year was also reduced by ACV treatment (70% placebo subjects vs 12.5% ACV subjects).(ABSTRACT TRUNCATED AT 250 WORDS)

Acyclovir↗
PubMed1984

The chromosomal location of mouse interferon alpha genes.

The chromosomal location of mouse leukocyte-interferon (IFN-alpha) genes was determined by Southern blot analysis of DNA from a panel of Chinese hamster x mouse somatic cell hybrids using a mouse IFN-alpha cDNA as a hybridization probe. All resolvable mouse genes are located on mouse chromosome 4. In addition, two common restriction site polymorphisms within these genes were identified in several mouse strains.

Animals↗
PubMed1983

Treatment of first episodes of genital herpes simplex virus infection with oral acyclovir. A randomized double-blind controlled trial in normal subjects.

We performed a double-blind placebo-controlled trial of oral acyclovir in the treatment of first episodes of genital herpes simplex virus infections in 48 young adults (31 women and 17 men). Subjects were randomized to receive either placebo or acyclovir (200 mg per dose) five times daily for 10 days; they were examined on at least eight visits until healed and at monthly visits thereafter. Acyclovir treatment, as compared with placebo, significantly reduced virus shedding, new lesion formation after 48 hours, and the duration of genital lesions in both men and women. The total duration and severity of clinical symptoms (such as pain, adenopathy, dysuria, and malaise) were significantly reduced by acyclovir in both men and women by the third and fourth day, respectively (P less than or equal to 0.025), as compared with placebo. No toxicity was observed. Recurrence rates have so far been similar in placebo and acyclovir recipients. Oral acyclovir treatment of first-episode genital herpes simplex virus infections is clinically effective, but it does not seem to prevent virus latency or associated recurrent disease.

Acyclovir↗
PubMed1982

Structure and synthesis of a simian virus 40 super T-antigen.

Mouse cells transformed by simian virus 40 often contain virus-coded tumor antigens distinct from those synthesized in productively infected permissive cells. The SV3T3 C120 cell line produces no large T-antigen of apparent molecular weight 94,000 but instead a super T-antigen of apparent molecular weight 145,000. We used recombinant DNA techniques to isolate the template for this super T-antigen and determined its structure by DNA sequencing. The integrated viral early transcription unit contains an in-phase, perfect tandem duplication of 1,212 base pairs. Transfer hybridization and endonuclease S1 mapping experiments were performed to elucidate the structures of the stable, cytoplasmic mRNAs of SV3T3 C120 cells, mRNAs of 3.9 and 3.6 kilobases, containing the small t- and large T-antigen splices, respectively, were transcribed from the internally duplicated early transcription unit. We showed by in vitro translation that these mRNAs encode small t-antigen and the super T-antigen of molecular weight 145,000. Peptide mapping studies of the SV3T3 C120 super T-antigen were consistent with its being derived from an internally duplicated template, since the protein has methionine and cysteine tryptic fingerprints virtually identical to those of normal large T-antigen, with certain methionine peptides present in greater than one molar yield.

Amino Acid Sequence↗
PubMed1982

Functional analysis of a simian virus 40 super T-antigen.

The SV3T3 C120 line of simian virus 40-transformed mouse cells synthesizes no large T-antigen of molecular weight 94,000 but instead a super T-antigen of molecular weight 145,000. In the accompanying paper (Lovett et al., J. Virol. 44:963-973, 1982), we showed that the integrated viral DNA segment SV3T3-20-K contains a perfect, in-phase, tandem duplication of 1.212 kilobases within the large T-antigen coding sequences. Our data suggested that this integrated template encodes mRNAs of 3.9 and 3.6 kilobases, the smaller of which directs the synthesis of the super T-antigen of molecular weight 145,000. We transfected the DNA segment SV3T3-20-K into nonpermissive rat cells and into TK- mouse L cells and analyzed the T-antigens and viral mRNAs in the transfectants; these data prove directly the coding assignments suggested previously. The super T-antigen retained the ability to induce morphological transformation, and may even transform better than the wild-type protein. It also retained the ability to bind to the cell-coded p53 protein. Transfection into permissive CV-1 cells showed that the super T-antigen encoded by SV3T3-20-K was incapable of initiating DNA replication at the viral origin. The duplication in SV3T3-20-K thus defines a mutation which separates the transformation and DNA replication functions of large T-antigen. We discuss why such mutations may be selected in transformed cells.

Animals↗
PubMed1980

The structure and expression of the integrated viral DNA in mouse cells transformed by simian virus 40.

Mouse cells stably transformed by simian virus 40 (SV40) contain viral DNA covalently integrated into their chromosomal DNA. We have used molecular cloning techniques to isolate and characterize the integrated viral DNA, together with the flanking cellular sequences, from several lines of transformed cells. We have identified in SV40-infected mouse cells a novel form of viral DNA, which we believe to be the precursor to the integrated DNA. The mRNAs transcribed from the integrated viral DNA templates have been characterized. Integrating the viral genome into the cellular chromosome can significantly alter its transcription pattern and in several cases the altered mRNAs encode novel forms of viral tumour antigen.

Animals↗