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L Duncan

Publications and source records attributed to L Duncan.

At least 55 records · Page 3Linked to original sources
PubMed1995

Activation of cell-specific transcription by a serine phosphatase at the site of asymmetric division.

Cell fate is determined by cell-specific activation of transcription factor sigma F after asymmetric division during sporulation by Bacillus subtilis. The activity of sigma F is governed by SpoIIAA, SpoIIAB, and SpoIIE, a membrane protein localized at the polar septum. SpoIIAB binds to and inhibits sigma F, and SpoIIAA inhibits SpoIIAB, which prevents SpoIIAB from binding to sigma F. SpoIIAB is also a serine kinase that inactivates SpoIIAA. Here, it is demonstrated that SpoIIE dephosphorylates SpoIIAA-P and overcomes SpoIIAB-mediated inhibition of sigma F. The finding that SpoIIE is a serine phosphatase links asymmetric division to the pathway governing cell-specific gene transcription.

Adenosine Triphosphate↗
PubMed1995

Synthetic oligonucleotide cocktails as probes for detection of human parvovirus B19.

A cocktail of 10 oligonucleotides selected at intervals along the length of the genome of human parvovirus B19 was labelled enzymically with digoxigenin and chemically with either digoxigenin (DIG) or dinitrophenyl (DNP). Chemical labelling was easier and more practical for the production of large quantities of probe. Pools labelled with either digoxigenin or DNP could detect 10 fg of B19 DNA in a dot blot reaction using an alkaline phosphatase antibody conjugate and colorimetric detection. Formalin fixed tissue from 11 consecutive cases of fetal hydrops were examined by in situ hybridisation (ISH). Both probe cocktails detected human parvovirus B19 DNA in 3 cases, with positive cells in all tissues examined and with equal sensitivity. The DNP pool is significantly cheaper and simpler to produce and could provide an inexpensive reagent suitable for diagnostic detection of viral nucleic acid in histopathological material.

Base Sequence↗
PubMed1995

Aromatase activity and interleukin-6 production by normal and malignant breast tissues.

The aromatase enzyme complex, which regulates the conversion of androstenedione to estrone, may have an important role in regulating estrogen synthesis in breast tissues. In this study the effect of tumor location on aromatase activity in adjacent tissue was examined and related to interleukin-6 (IL-6) production, which has been shown to stimulate aromatase activity in breast cancer cells. Samples of normal and malignant breast tissues were obtained from 11 women undergoing mastectomy. In 7 patients, aromatase activity was highest in the quadrant in which the tumor was located or on which the tumor impinged. Aromatase activity in tumor-bearing quadrants was significantly higher than that in adjacent and opposite quadrants. Aromatase activity and IL-6 production, expressed in terms of tissue weight, were significantly higher for tumor tissue compared with normal breast adipose tissue. A significant correlation was found between aromatase activity and IL-6 production for breast tumor tissue (rs = 0.56; P < 0.05), but not for adipose tissue from the breast quadrants. Aromatase activity and IL-6 production were also measured in tissue obtained from a normal woman undergoing reduction mammoplasty who had previously had breast augmentation by silicone injection, not contained within a capsule. In tissue from this patient there was evidence of chronic inflammation and a marked macrophage response. Aromatase activity in this tissue was considerably higher than that detected in mastectomy adipose tissue samples, and a significant correlation was found between aromatase activity and IL-6 production (rs = 0.77; P < 0.05). A preliminary study to examine the potential role of cells of the immune system in regulating breast tissue aromatase activity revealed that conditioned medium collected from macrophages and lymphocytes could markedly stimulate aromatase activity in tumor-derived fibroblasts. The results of this study confirmed that breast tumor location can influence aromatase activity in adjacent tissues and showed that aromatase activity is increased in tumor-bearing quadrants. The increased production of IL-6 by tumor tissue and its correlation with aromatase activity suggest that tumors may be the major source of IL-6, which is able to influence aromatase activity in adjacent tissues.

Adipose Tissue↗
PubMed1994

An adenosine nucleotide switch controlling the activity of a cell type-specific transcription factor in B. subtilis.

The sigma F factor establishes cell type-specific gene transcription during sporulation in B. subtilis. sigma F is negatively regulated by SpollAB, which forms complexes with sigma F or SpollAA. ATP and its nonhydrolyzable analogs stimulate the formation of the SpollAB.sigma F complex, whereas ADP stimulates the formation of the SpollAB.SpollAA complex. Which protein SpollAB associates with is determined by the concentrations of the two nucleotides, on which basis we propose a partner-switching model for the regulation of sigma F: [formula: see text] Consistent with this model, SpollAA reverses SpollAB-mediated inhibition of sigma F-directed transcription in a manner that depends on ADP. Cell-specific activation of sigma F could be due to an alteration in adenosine nucleotide levels in one cell of the sporangium.

Adenosine Diphosphate↗
PubMed1994

Establishment of cell type specific gene transcription during sporulation in Bacillus subtilis.

Asymmetric cell division during the process of sporulation in the bacterium Bacillus subtilis generates dissimilar progeny that exhibit distinct programs of gene transcription. Recent work reveals a partner switching mechanism that governs the activity of the sporulation regulatory protein sigma F and that may be responsible for the establishment of cell type specific gene transcription.

Adenine Nucleotides↗
PubMed1993

SpoIIAB is an anti-sigma factor that binds to and inhibits transcription by regulatory protein sigma F from Bacillus subtilis.

The sigma F factor is a regulatory protein that is responsible for directing gene expression in the forespore compartment of developing cells of the spore-forming soil bacterium Bacillus subtilis. The sigma F factor is encoded by the promoter-distal member of sporulation operon spoIIA, which consists of cistrons called spoIIAA, spoIIAB, and spoIIAC. Genetic evidence indicates that the activity of sigma F is negatively regulated by the product (SpoIIAB) of the spoIIAB cistron. We now report that SpoIIAB is capable of binding to sigma F and inhibiting its capacity to direct transcription by core RNA polymerase from the promoter for a forespore-expressed gene. SpoIIAB is an anti-sigma factor that may be directly involved in the compartmentalization of sigma F-directed gene expression.

Bacillus subtilis↗
PubMed1993

Inhibition of estrone sulfatase activity by estrone-3-methylthiophosphonate: a potential therapeutic agent in breast cancer.

Many breast tumors are hormone dependent, and there is evidence that hydrolysis of estrone sulfate (E1S) to estrone, by estrone sulfatase, is an important source of the estrogen which is found in tumors. In this study, we have developed a novel pathway for the synthesis of estrone-3-methylthiophosphonate (E1-3-MTP) and examined its ability to inhibit estrone sulfatase activity in MCF-7 breast cancer cells and human placental and breast tumor preparations. In MCF-7 breast cancer cells, E1-3-MTP, 100 nM and 10 microM, inhibited estrone sulfatase activity by 52 and > 98%, respectively. The apparent Km and Vmax for E1S were 4.8 microM and 148 pmol/min/mg for placental and 16.9 microM and 38 pmol/min/mg for breast tumor preparations. Kinetic studies revealed that E1-3-MTP inhibited estrone sulfatase in a competitive manner with the Ki values for placental and tumor preparations being 14.6 and 32.8 microM, respectively. A comparison of the metabolism of [3H]E1S and [3H]E1-3-MTP by human placenta or rat liver revealed that, whereas 50-60% of [3H]E1S was converted to [3H]estrone, < 3% of [3H]E1-3-MTP was hydrolyzed. The development of an efficient inhibitor of estrone sulfatase, which is resistant to metabolism, will allow the importance of the estrone sulfatase pathway of estrogen formation in breast tumors to be assessed and such an inhibitor may have considerable potential as a therapeutic agent.

Animals↗
PubMed1993

Co-amplification of the gene for parathyroid hormone-related protein (PTHRP) and KRAS2 in a human lung cancer cell line.

Parathyroid hormone-related protein (PTHRP) is expressed in a large number of tumors and is the mediator of parathyroid hormone-like effects seen in humoral hypercalcemia of malignancy. The gene coding for PTHRP has been localised to the short arm of chromosome 12. This is at the same region as the oncogene KRAS2, and amplification of KRAS2 has previously been found in human lung cancer. The BEN cell line which is known to express PTHRP was established from a patient who had squamous cell carcinoma of the lung with hypercalcemia. Cytogenetic analysis of the BEN cell line revealed a very complex karyotype with many marker chromosomes. Chromosomal in situ hybridization with biotinylated DNA probes visualized by a biotin-streptavidin-polyalkaline-phosphatase complex was used to analyse two dicentric marker chromosomes containing homogeneously staining regions (hsr) in BEN. The hsr were found to contain amplified PTHRP and KRAS2 at levels of 30-fold and 14-fold per cell, respectively. The higher level of amplification of the PTHRP gene would suggest that PTHRP is the target gene of amplification in the amplicon. This is the first report of gene amplification of PTHRP and in addition its co-amplification with KRAS2.

Chromosome Aberrations↗
PubMed1993

Sexual function in women. Do antihypertensive drugs have an impact?

Drug-induced sexual dysfunction is well known to occur with antihypertensive drugs in men. There are much less data on the effects of drugs on female sexual function. The physiology of the female sexual response has similarities to that of the male sexual response and there are therefore good reasons for suspecting that antihypertensive drugs are likely to adversely affect sexual function in women. Present evidence suggests that clonidine, methyldopa, guanethidine and reserpine are associated with adverse effects on sexual function. In healthy volunteers, labetalol appears to reduce vaginal lubrication, but there are no studies in patients receiving the drug therapeutically. Thiazide diuretics may be associated with the worsening of sexual problems, which interestingly appear to be ameliorated by weight reduction. Present evidence on the effects of vasodilators is limited but the evidence suggests that sexual function in women receiving calcium antagonists is not altered by changing to an angiotensin converting enzyme (ACE) inhibitor. Although present evidence suggests that effects on female sexual function may not be very great, it should be recognised that there are very few data in this area. Further work is clearly necessary.

Antihypertensive Agents↗
PubMed1992

Modulation of oestrone sulphatase activity in breast cancer cell lines by growth factors.

Currently there is much interest in the role that growth factors may play in the development of human breast tumours. We have shown previously that growth factors secreted by breast tumours may influence the activity of oestradiol hydroxysteroid dehydrogenase, the enzyme which catalyses the interconversion of oestrone (E1) and oestradiol. As the formation of E1 from its sulphate (E1S) by oestrone sulphatase may be quantitatively more important than production from androstenedione via aromatase, we have studied the effect of insulin-like growth factor-1 (IGF-I) and basic fibroblast growth factor (bFGF) on oestrone sulphatase activity in the hormone-dependent MCF-7 and the hormone-independent MDA-MB-231 breast cancer cell lines. In both these cell types, bFGF (1-200 ng/ml) and IGF-I (25-200 ng/ml) significantly stimulated oestrone sulphatase activity in a dose-dependent manner (by 8-60%) after 48 h. Additionally, cycloheximide significantly inhibited (by 90-120%) this stimulation of oestrone sulphatase activity by the two growth factors in both MCF-7 and MDA-MB-231 cells. Basal oestrone sulphatase activity was higher in the oestrogen receptor, ER-ve MDA-MB-231 cells than in the ER + ve MCF-7 breast cancer cells. We conclude that these growth factors, believed to be secreted by breast tumours, may induce enzymes of oestrogen synthesis and hence increase local production of oestrogens.

Breast Neoplasms↗
PubMed1991

Glomus tumor of the coccyx. A curable cause of coccygodynia.

A 30-year-old woman presented with recurrent severe coccygodynia. She underwent exploration for a possible pilonidal sinus and was found to have a precoccygeal glomus tumor that also involved bony trabeculae of the coccyx. To our knowledge, a glomus tumor involving the coccygeal bone has not been previously documented. In view of the relief of this patient's pain following the surgical excision of coccyx and tumor, a causal role is suggested.

Adult↗
PubMed1990

The synthesis of polyamide-oligonucleotide conjugate molecules.

We have developed methods for the synthesis of peptide-oligodeoxyribonucleotide conjugate molecules in particular, and polyamide-oligonucleotide conjugates in general. Synthesis is carried out by a solid-phase procedure and involves the assembly of a polyamide on the solid support, conversion of the terminal amino group to a protected primary aliphatic hydroxy group by reaction with alpha, omega-hydroxycarboxylic acid derivatives, and finally oligonucleotide synthesis using phosphoramidite chemistry. The conjugate molecules can be used as DNA probes, with the polyamide component carrying one or more non-radioactive markers. These conjugates also have the potential to be used as anti-sense inhibitors of gene expression, with the peptide segment acting as a targeting moiety.

Amino Acid Sequence↗
PubMed1990

The preparation of polyamide-oligonucleotide probes containing multiple non-radioactive labels.

Oligonucleotide probes containing multiple non-radioactive labels have been prepared by utilising and extending the methods used to prepare polyamide-oligonucleotide conjugates. The probes were prepared by incorporating suitable amino acid residues, such as lysines, in the polyamide, which were then used as sites for the attachment of the non-radioactive labels. The procedures developed give control over the distance of the label from the oligonucleotide, and also the inter-label distance. The labels can be conveniently introduced while the substrate is still on the solid support. Even though fluorescent oligonucleotide probes prepared in this way carrying multiple carboxyfluorescein labels gave low levels of fluorescence due to quenching, the probes containing ten biotin labels gave a detection sensitivity of approximately 5 attomole (3 million molecules).

Animals↗
PubMed1990

Control of developmental transcription factor sigma F by sporulation regulatory proteins SpoIIAA and SpoIIAB in Bacillus subtilis.

The sporulation operon spoIIA of Bacillus subtilis consists of three cistrons called spoIIAA, spoIIAB, and spoIIAC. Little is known about the function of spoIIAA and spoIIAB, but spoIIAC encodes a sigma factor called sigma F, which is capable of directing the transcription in vitro of genes that are expressed in the forespore chamber of the developing sporangium. We now report that the products of the spoIIA operon constitute a regulatory system in which SpoIIAA is an antagonist of SpoIIAB (or otherwise counteracts the effect of SpoIIAB) and SpoIIAB is, in turn, an antagonist of SpoIIAC (sigma F). This conclusion is based on the observations that (i) overexpression of spoIIAB inhibits sigma F-directed gene expression, (ii) a mutation in spoIIAB stimulates sigma F-directed gene expression, (iii) a mutation in spoIIAA blocks sigma F-directed gene expression, and (iv) a mutation in spoIIAB relieves the block in sigma F-directed gene expression caused by a mutation in spoIIAA. The SpoIIAA/SpoIIAB/SpoIIAC regulatory system could play a role in controlling the timing of sigma F-directed gene expression and/or could be responsible for restricting sigma F-directed gene expression to the forespore chamber of the sporangium.

Bacillus subtilis↗