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Biomedical subjects

M Page

Publications and source records attributed to M Page.

At least 109 records · Page 6Linked to original sources
PubMed1986

A mechanism for the insertion of complement component C9 into target membranes.

Complement component C9 is a globular serum protein which can insert and polymerise in a target membrane to form a large membrane channel. The ability to insert in the membrane is conferred by amphipathetic elements of secondary structure in the central part of the molecule. Towards each end high cysteine domains are found, one of which is homologous to the apoprotein binding domains of the LDL receptor. From the sequence and topological data for C9 we present a model for its structure and insertion into the membrane.

Cell Membrane↗
PubMed1986

Androgen regulation by the long terminal repeat of mouse mammary tumor virus.

Mouse mammary tumor virus (MMTV) has long been implicated in mouse mammary carcinogenesis, and it is now well established that the long terminal repeat (LTR) contains regulatory sequences responsible for glucocorticoid-mediated induction of viral RNA. However, we have demonstrated previously that androgens as well as glucocorticoids can regulate MMTV RNA in the S115 mouse mammary tumor cell line. To determine if androgens act directly on the LTR in these cells, plasmids were constructed with the MMTV LTR joined to the coding sequences of genes not normally expressed in the cells. Following transfection of these chimeric genes into S115 cells, we show that the expression of the genes is regulated by both androgens and glucocorticoids. Furthermore, hormonal regulation is also conferred by the LTR on the neighboring guanine phosphoribosyltransferase (gpt) gene. Thus, androgens can act on the LTR of MMTV when the appropriate receptors are present in the cells, and this interaction can influence the expression of additional adjacent genes.

Androgens↗
PubMed1985

Transferrin and iron release from rat hepatocytes in culture.

The regulation of transferrin and iron release from the liver was studied using adult rat hepatocytes in primary monolayer culture. The cells were prelabeled by incubation with rat transferrin doubly labeled with iodine-125 and iron-59. Approximately 50% of the 125I-transferrin but only 10% of the iron-59 taken up by the cells was released during reincubation for 24 h. Less than 10% of the refluxed transferrin was catabolized as indicated by the protein-free iodine-125 values. These results suggest that at least part of iron uptake by hepatocytes is mediated by the reversible binding of transferrin in a manner comparable with erythroid cells and placenta. However, several iron chelators mobilized hepatic iron, in contrast to erythroid cells. Apotransferrin and desferrioxamine released a maximum of about 20% iron-59 with little effect on transferrin binding. A greater proportion of the iron-59 was available for chelation after shorter uptake times (1-2 h) than longer times. Hence, there are at least three iron compartments in hepatocytes in culture: rapidly refluxing iron that may be transferrin bound, a fixed pool, and a chelatable pool that may represent iron in transit between plasma transferrin and ferritin.

Animals↗
PubMed1985

Effect of desferrioxamine, rhodotorulic acid and cholylhydroxamic acid on transferrin and iron exchange with hepatocytes in culture.

The mechanism of action of the hydroxamate iron chelators desferrioxamine (DFO), rhodotorulic acid (RHA) and cholylhydroxamic acid (CHA) was studied using rat hepatocytes in culture. Each chelator affected both the uptake and, to a much smaller extent, the release of transferrin-125I-59Fe from the cells. All chelators reduced the 59Fe uptake and incorporation into ferritin in a concentration-dependent manner. Uptake of 59Fe into the membrane (stromal-mitochondrial) fraction was also decreased by DFO and RHA but increased by CHA. Transferrin-125I binding was reduced slightly by DFO and RHA and increased by CHA. All chelators released 59Fe transferrin-125I from hepatocytes prelabelled by incubation with rat transferrin-125I-59Fe and washed before reincubation in the presence of the chelators. DFO decreased membrane 59Fe but had little effect on ferritin-59Fe. RHA decreased 59Fe in both membrane and ferritin fractions. CHA decreased hepatocyte-59Fe but increased 59Fe in the hepatocyte membrane fraction. Higher concentrations of the chelators had little further effect on 59Fe release but promoted transferrin-125I release from hepatocytes. All chelators appeared to act on kinetically important iron pools of limited size and hence are likely to be most effective when given by continuous infusion rather than bolus injection.

Animals↗
PubMed1984

Fluorescence determination of microconcentrations of chlorambucil after photoactivation.

A fluorescence assay is described which measures the alkylating activity of chlorambucil or its isocyanate derivative after photoactivation in the presence of dimethyl sulfoxide. This assay has a lower limit of sensitivity of 100 ng/mL and RSD of less than 10% for chlorambucil. The method requires less than 5 micrograms of alkylating agent and the fluorophore produced is stable for at least 24 h.

Chlorambucil↗
PubMed1984

Organization and expression of prostatic steroid binding protein genes.

The function of regulatory regions of DNA, which flank the genes for prostatic steroid binding protein, is being analysed by introducing the cloned genes into heterologous cells. Two non-allelic genes for the C3 polypeptide, C3(1) and C3(2) have been transfected into androgen-responsive S115 cells using SV2-gpt vectors. The genes have been introduced either intact or as so-called fusion genes consisting of putative C3 promoters plus human beta-interferon cDNA. Both genes for C3 were accurately transcribed and their expression was stimulated 5-10-fold with 10(-8) M testosterone. Thus we should be able to define at least one region of DNA which confers androgen sensitivity to the genes. Although both genes were expressed similarly in S115 cells only C3(1) is responsible for C3 in vivo and C3(2) is transcribed poorly, if at all. The most likely explanation for the difference is that in vivo the C3(2) gene remains hypermethylated whereas the DNA which was introduced into S115 cells was unmethylated. This result supports the notion that the state of DNA methylation is important for controlling the transcription of genes.

Androgen-Binding Protein↗
PubMed1984

Organization and expression of genes encoding prostatic steroid binding protein.

We have cloned the genes for prostatic steroid binding protein to study the mechanism whereby their expression is regulated by testosterone. The genes for the C1 and C2 polypeptides are probably unique whereas there are two genes C3(1) and C3(2) for the C3 polypeptide of which only the former is transcribed in vivo. The state of DNA methylation associated with the two genes for C3 also differ, insofar as C3(1) is demethylated in ventral prostate from 14-28 days of age, whereas the C3(2) gene remains hypermethylated. The organization of all four genes is similar and appreciable DNA sequence homologies suggest that they may have arisen from a single ancestral gene. To study C3 expression and its hormonal regulation we have introduced the cloned genes into mouse S115 cells, an androgen-responsive cell line. Both genes were accurately transcribed and their expression was stimulated up to fivefold by 10(-8) M testosterone in approximately one third of the clones tested. To delineate the site of action of the hormone we have constructed chimeric genes consisting of putative C3 promoters and regulatory sequences together with a marker gene, interferon. This chimeric gene resulted in interferon production but its expression was stimulated by less than twofold in all clones tested. Therefore, these results indicate that, in mouse cells, testosterone does not interact directly with the rat C3 promoter but, in certain clones, may act post-transcriptionally.

Androgen-Binding Protein↗
PubMed1983

Growth modification of normal and tumor cell lines with retinol.

The behavior of some normal and tumor cell lines was studied after treatment with retinol. Human amnion CCL-25 cells, human embryonic intestine CCL-6 cells, human colon adenocarcinoma LOVO cells, and mouse hepatoma BW-1 cells were used. Acute and chronic toxicity of vitamin A was evaluated. The effect of retinol on cell proliferation was monitored with regard to the cell growth rate and the number of cells at saturation density. The influence of the metabolic status of the cell was studied with respect to its susceptibility to antiproliferative effect of retinol. Major effects on growth inhibition could be observed with BW-1 and CCL-6 cells, especially when they were seeded at low density.

Adenocarcinoma↗
PubMed1982

Prostatic steroid binding protein: organisation of C1 and C2 genes.

Prostatic steroid binding protein, whose expression is stimulated by androgens, consists of two subunits: one containing the polypeptides C1 and C3 and the other containing C2 and C3. We have characterised genomic clones containing the C1 and C2 genes by restriction enzyme analysis and DNA sequencing. Both genes are 3.2 Kb, have similar exon/intron arrangements and share considerable DNA sequence homologies in their coding regions, intervening sequences and 5' upstream DNA sequence which suggests that they have probably arisen from the duplication of an ancestral gene. The 5' termini of C1 and C2 mRNA have been mapped; the sequence TATAAA appears 30 nucleotides upstream but a CAAT-like sequence at -60 - -80 is absent. Finally, homologous human genes have not been detected.

Androgen-Binding Protein↗