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Biomedical subjects

M Braddock

Publications and source records attributed to M Braddock.

43 records · Page 3Linked to original sources

HIV-1 TAT "activates" presynthesized RNA in the nucleus.

Replication of the human immunodeficiency virus (HIV-1) depends upon the viral TAT protein. TAT stimulates gene expression via a target response sequence (TAR) located within the HIV-1 LTR. As TAR is located in the transcribed region it could act as a signal in either the DNA, the RNA, or both. To test whether TAT acts on transcription and/or posttranscriptionally, we produced TAT in yeast and monitored its activity after microinjection into the nucleus or cytoplasm of Xenopus oocytes. The TAT protein stimulated TAR-dependent expression, but this activation was not inhibited by transcriptional inhibitors. Furthermore, TAR-containing RNA, produced in vitro, was "activated" by TAT after coinjection into oocytes. This activation only occurred, however, when the RNA was injected into the nucleus and not into the cytoplasm. Our data indicate, therefore, that in the Xenopus system TAT acts on presynthesized RNA and that the nucleus is involved in this action.

Animals↗

HIV expression strategies: ribosomal frameshifting is directed by a short sequence in both mammalian and yeast systems.

The pol gene of the human immunodeficiency virus (HIV-1) is expressed as a gag:pol fusion, arising from a ribosomal frameshift that brings the overlapping, out-of-phase gag and pol genes into translational phase. In this study, we show that HIV frameshifting is mediated by a very short sequence in the viral RNA. We demonstrate the importance of a homopolymeric run within this sequence and conclude that HIV frameshifting is not dependent on stem-loop structures downstream from the frameshift site. Our analysis also indicates that the sequence requirements are identical in mammalian and yeast systems.

Animals↗

Synthesis of a gene for the HIV transactivator protein TAT by a novel single stranded approach involving in vivo gap repair.

The synthesis of a gene for the HIV TAT protein is described using a novel approach that capitalises on the ability to synthesise oligonucleotides of greater than 100 bp in length. It involves the synthesis of large oligomers covering one strand of the desired gene in its entirety and the use of small complementary bridging and adapter oligonucleotides to direct the assembly and cloning of the large oligomers. After ligation to the cloning vector the partially single stranded intermediate is transformed directly into the recipient bacterial host where the plasmid is repaired. The synthetic tat gene has been expressed in HeLa cells and is shown to trans-activate TAR+ but not TAR- HIV LTR-CAT constructs.

Amino Acid Sequence↗

Cloning of cDNA to rat mammary-gland fatty acid synthase mRNA. Evidence for the expression of two mRNA species during lactation.

A cDNA library was constructed in the expression vector lambda gt11, by synthesizing cDNA from size-selected poly(A) RNA from lactating rat mammary gland, using random hexanucleotide primers. Using this library we identified two recombinants which, on addition of a lac z inducer, produced proteins recognized by affinity-purified anti-fatty-acid synthase antibody, and which, therefore, contained fatty acid synthase coding sequences. The inserts were subcloned, were shown to be between 500 and 600 base pairs in size, and to cross-hybridize. The cloned DNA was then used in Northern hybridizations with mRNA isolated at various stages throughout lactation. Two mRNA species were identified of approx. 9.7 and 10.4 kilobases, which increased and decreased in parallel during lactation, reaching a peak at 12-13 days. Both mRNA species disappeared rapidly if the pups were removed prematurely. This study provides evidence that, during hormonal induction in lactation, regulation of the level of fatty acid synthase protein can be accounted for by variation in the level of mRNA.

Animals↗

Medical assessment of children going into emergency out-of-home placement.

Approximately 500,000 children and infants go into out-of-home placement in the United States each year. Increased attention is being given to the health care of children as they enter and remain in placement. This article describes a model, which has been in operation for 5 years, that provides medical assessments of children as they enter emergency out-of-home placement. The model is a community partnership with the county social service department, the police department, and the hospital. A computerized database that contains records for each child, including medical findings, has been helpful in developing a profile of the children served and contributes to continuity of care.

Adolescent↗

Variations in pregnancy outcomes by race among 10-14-year-old mothers in the United States.

This study used the 1983-86 U.S. Linked Live Birth-Infant Death Files to examine variations in pregnancy outcomes among 38,551 U.S. resident black and white adolescents ages 10 through 14. The birth rate was 4.29 per 1,000 for blacks, more than 7 times the rate for whites (.59 per 1,000). Black mothers had higher proportions of very low and low birth weight infants than did whites (very low birth weight: 3.7 versus 2.6; low birth weight: 15.0 versus 10.5). Neonatal and infant mortality rates were higher among very low birth weight and low birth weight white infants. Neonatal and infant mortality rates were similar for normal birth weight infants of both races, but were 3.7 to 7.4 times higher among black infants with birth weights more than 4,250 grams. Logistic regression indicated that black mothers were at higher risk for having infants who were low birth weight, very low birth weight, small for gestational age, preterm, and very preterm. There were no differences by race for neonatal, postneonatal, and infant mortality. While the risk for poor pregnancy outcomes is great among young adolescents, young black adolescents appear to be particularly vulnerable. Attempts to reduce unintended pregnancies in this group should receive highest priority.

Adolescent↗