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Biomedical subjects

L J Nicholson

Publications and source records attributed to L J Nicholson.

13 recordsLinked to original sources
PubMed1999

A20 RNA expression is associated with undifferentiated nasopharyngeal carcinoma and poorly differentiated head and neck squamous cell carcinoma.

A20 is an anti-apoptotic gene that can be induced in human epithelial cell lines in response to expression of the Epstein-Barr virus (EBV) gene product latent membrane protein 1 (LMP1). EBV is a ubiquitous, persistent human herpesvirus that is consistently associated with undifferentiated nasopharyngeal carcinoma (NPC), in which antigen expression includes LMP1. Consistent with a potential role in the development of NPC, LMP1 has profound effects on epithelial cell growth. A20 may be a key downstream effector of LMP1 in NPC, as LMP1-induced A20 blocks p53-mediated apoptosis in H1299 epithelial cells and most NPCs have wild-type p53. Moreover, the potential role of A20 in the development of epithelial malignancies may extend to tumours not associated with EBV. The purpose of this study was to develop an in situ hybridization assay to assess expression of A20 RNA in undifferentiated NPC and in non-EBV-associated poorly differentiated head and neck squamous cell carcinomas (SCCs) and well-differentiated SCCs of the skin. A20 RNA expression was also examined in normal samples of oral mucosa and skin. Expression of A20 was demonstrated in 76 per cent of undifferentiated NPCs and in 80 per cent of poorly differentiated head and neck SCCs, suggesting a role for A20 in the pathogenesis of these epithelial malignancies. By contrast, A20 RNA was not detected in well-differentiated SCCs of the skin, or in any normal samples of squamous epithelial tissue. The pathway leading to A20 expression in non-EBV-associated poorly differentiated head and neck SCCs is clearly LMP1-independent. LMP1 expression was demonstrated in 29 per cent of NPC biopsies, suggesting an LMP1-independent pathway to A20 induction in undifferentiated NPC.

Animals↗
PubMed1999

Definition of the transcription factors which bind the differentiation responsive element of the Epstein-Barr virus BZLF1 Z promoter in human epithelial cells.

Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus and an important human pathogen. Initiation of the EBV lytic cycle is dependent upon transcription of the EBV BZLF1 gene. Our previous studies of transcriptional regulation of the BZLF1 Z promoter (Zp) in human SCC12F epithelial cells identified a region within Zp that is responsive to epithelial cell differentiation. In the present study, we localize this differentiation responsive element to the CREB/AP-1-like binding site (TGACATCA) between -67 to -60 bp within Zp, previously designated ZII, and furthermore show that homodimers and heterodimers of CREB and ATF-1 specifically bind ZII. Consistent with a regulatory role for CREB and ATF-1 in differentiation dependent BZLF1 expression, ZII was able to bind approximately 3-fold more CREB and ATF-1 when incubated with nuclear extract obtained from populations of SCC12F cells enriched for the differentiated phenotype than when incubated with extract obtained from populations enriched for the undifferentiated phenotype. In addition, CREB and ATF-1 were found to increase in abundance during SCC12F differentiation. These results indicate a regulatory role for CREB and ATF-1 in differentiation-dependent expression of BZLF1 in human epithelial cells.

Activating Transcription Factor 1↗
PubMed1997

Epstein-Barr virus latent membrane protein does not inhibit differentiation and induces tumorigenicity of human epithelial cells.

Latent membrane protein (LMP) is a latent Epstein-Barr virus (EBV) protein expressed in the EBV associated malignancy, nasopharyngeal carcinoma (NPC). Properties ascribed to this protein include inhibition of epithelial cell differentiation and deregulation of epithelial cellular gene expression, and are believed to contribute to the development of NPC. Studies to evaluate the oncogenic potential of LMP in epithelial cells have not been conclusive. We carried out studies to determine the tumorigenic activity of LMP in two human epithelial cell lines, SCC12F and HaCaT; while SCC12F LMP transfectants were non-tumorigenic in severe combined immunodeficient mice, HaCaT LMP transfectants were strongly oncogenic. The tumours produced were well differentiated, keratinising squamous cell carcinomas suggesting that LMP does not inhibit epithelial cell differentiation which conflicts with a previous report by Dawson et al. (1990). To resolve this discrepancy we examined the ability of HaCaT and SCC12F LMP transfectants to differentiate in a suspension culture assay. Both lines were able to differentiate to a similar extent as parental lines and control transfectants. Our results indicate that LMP is strongly oncogenic in human epithelial cells but that inhibition of differentiation is not necessarily a mechanism by which LMP contributes to the pathogenesis of NPC.

Animals↗
PubMed1995

Identification of an epithelial cell differentiation responsive region within the BZLF1 promoter of the Epstein-Barr virus.

The Epstein-Barr virus (EBV) BZLF1 gene product, Zta is able to trigger the viral lytic cycle in latently infected B lymphocytes. Investigations into regulation of the promoter, Zp for the BZLF1 gene in B cells have identified 12-O-tetradecanoylphorbol 13-acetate, anti-immunoglobulin and Zta responsive elements as well as a negative regulatory element within Zp. EBV infects and replicates within squamous epithelial cells and there is evidence to indicate that EBV gene expression is linked to the differentiation status of epithelial cells. We have investigated regulation of Zp in undifferentiated and differentiated cells of the human squamous epithelial cell line SCC12F. Zp was found to be active in SCC12F cells and activity was increased approximately 7-fold upon induction of epithelial terminal differentiation. Sequences responsive to epithelial cell differentiation were contained within the region of Zp from -86 to +12 bp, a region previously shown to contain an AP-1-like binding site, designated the ZII domain. In addition, enhancing sequences were detected in the region -221 to -86 bp. These studies will lead to a greater understanding of differentiation-linked EBV gene expression in human squamous epithelial cells.

Animals↗
PubMed1993

Regulation of keratinocyte terminal differentiation by integrin-extracellular matrix interactions.

Suspension-induced terminal differentiation of human epidermal keratinocytes can be inhibited by fibronectin through binding to the alpha 5 beta 1 integrin. We have investigated the effect of fibronectin on expression of integrins and proteins of the actin cytoskeleton and have explored the nature of the differentiation stimulus by testing different combinations of anti-integrin monoclonal antibodies or extracellular matrix proteins in the suspension assay. Fibronectin prolonged cell surface expression of beta 1 integrins but did not overcome the inhibition of intracellular transport of integrins that occurs when keratinocytes are placed in suspension. Fibronectin did not prevent the suspension-induced decline in the level of mRNAs encoding the beta 1 integrin subunit, actin, filamin and alpha-actinin; furthermore, the inhibition of terminal differentiation did not depend on the state of assembly of microfilaments or microtubules. Terminal differentiation could be partially inhibited by an adhesion-blocking monoclonal antibody to the beta 1 integrin subunit or by a combination of adhesion blocking antibodies recognising the alpha subunits that associate with beta 1 (alpha 2, alpha 3 and alpha 5). Although laminin and type IV collagen do not inhibit terminal differentiation individually, they were inhibitory when added to cells in combination with a low concentration of fibronectin. We conclude that the proportion of keratinocyte beta 1 integrins occupied by ligand can regulate the initiation of terminal differentiation independently of the state of assembly of the actin cytoskeleton.

Actins↗
PubMed1991

Expression of E-cadherin, P-cadherin and involucrin by normal and neoplastic keratinocytes in culture.

We have compared expression of involucrin, E-cadherin and P-cadherin in cultures of normal keratinocytes and in five different lines derived from squamous cell carcinomas (SCCs), using Northern analysis and immunofluorescence. In normal keratinocytes there was an inverse correlation between P-cadherin and involucrin expression, whereas E-cadherin was expressed by both basal and terminally differentiating cells. In SCC lines involucrin expression was lower than in normal keratinocytes, and there was variable expression of P- and E-cadherin: E-cadherin mRNA levels tended to be lower in SCC lines than in normal keratinocytes, whereas P-cadherin levels were similar. Our results are consistent with observations of cadherin expression in vivo and suggest that the cultures provide a useful experimental model for investigating the role of cadherins in determining the spatial organization of normal and neoplastic keratinocytes.

Cadherins↗
PubMed1991

Decreased expression of fibronectin and the alpha 5 beta 1 integrin during terminal differentiation of human keratinocytes.

We have examined the expression of fibronectin and the alpha 5 beta 1 fibronectin receptor during terminal differentiation of human epidermal keratinocytes, using involucrin as a terminal differentiation marker. The levels of mRNAs encoding fibronectin and the alpha 5 and beta 1 integrin subunits were measured in keratinocyte populations that had been enriched for involucrin-negative or -positive cells by unit gravity sedimentation or suspension-induced terminal differentiation. All three mRNAs decreased in abundance during terminal differentiation, and the corresponding proteins were localised by immunofluorescence to the basal layer in stratified colonies. We also examined expression in ndk, a strain of epidermal cells with a complete block in terminal differentiation, which, as a result, do not express involucrin. Messenger RNA levels for fibronectin and the alpha 5 and beta 1 subunits were higher in ndk, than in unfractionated keratinocytes and the corresponding proteins were expressed by all ndk, consistent with a basal keratinocyte phenotype. We conclude that expression of fibronectin and the alpha 5 beta 1 fibronectin receptor decreases during terminal differentiation and that such changes are likely to play a role in the selective migration of terminally differentiating cells from the basal epidermal layer.

Antibodies, Monoclonal↗
PubMed1988

Effects of sodium cyclamate and sodium saccharin on focus induction in explant cultures of rat bladder.

The tumour-promoting activities of sodium cyclamate and sodium saccharin were investigated in an assay based on the induction of epithelial foci exhibiting enhanced growth potential in a rat bladder explant culture system. An initiating, non-focus-inducing dose was defined for the carcinogen N-methyl-N-nitrosourea (MNU) to make promotion studies possible. Saccharin induced epithelial foci when added to cultures pretreated with an initiating dose of MNU, and also increased the incidence of foci in cultures treated with transforming doses of MNU. Cyclamate was found to induce a high incidence of foci when added to cultures by itself. When MNU and cyclamate treatments were combined, an additive effect could be detected. These results indicate that both cyclamate and saccharin can contribute to epithelial transformation in this system.

Animals↗
PubMed1985

Cigarette smoke-induced injury of peritoneal mesothelial cells.

Cultured rat peritoneal mesothelial cells have been exposed to plasma obtained from healthy adult male volunteers before and after the smoking of two medium-tar cigarettes. The pre-smoking plasma had little effect on the mesothelial cells as observed by scanning electron microscopy. Post-smoking plasma produced extensive cellular damage evidenced by the production of blebs and microvillus-like structures on the cell surface. Some cells were also found to have a perforated luminal membrane. These results suggest that factors cytotoxic for these cells are present in plasma following exposure of humans to cigarette smoke.

Adult↗
PubMed1984

The mesothelial cell as a non-thrombogenic surface.

A technique for harvesting mesothelial cells is described. This entails collagenase digestion of omentum after which the cells can be cultured. The technique has been developed using the rat, but has also been successfully applied to human tissue. Cultured rat mesothelial cells obtained in this way have been examined by scanning electron microscopy. Rat mesothelial cells grown on plastic film have been exposed to blood in an in vitro system using a Baumgartner chamber and have been demonstrated to support blood flow. No adhering platelets were observed on the mesothelial cell surface. Fibroblasts similarly exposed to blood as a control were washed off the plastic.

Animals↗
PubMed1984

Seeding Dacron arterial prostheses with peritoneal mesothelial cells: a preliminary morphological study.

A preliminary laboratory study has been undertaken using dogs, in which porous Dacron arterial prostheses have been seeded with autologous peritoneal mesothelial cells before implantation into the arterial system. These cells were harvested from omentum by collagenase digestion and were introduced into the graft at the time of preclotting. Examination of the grafts by scanning and transmission electron microscopy one month after insertion showed no organized cellular lining in the control graft. In three seeded grafts there was a lining of mesothelial cells which extended over the whole surface of the grafts up to 1 cm from the suture lines. Selected areas of the grafts have been analysed morphometrically. In the seeded grafts there is a cellular layer which on average covers 94 per cent of the luminal surface. These results suggest that mesothelial cells, which both secrete prostacyclin and possess fibrinolytic activity, present a possible alternative to endothelium as a cellular lining for prosthetic grafts.

Animals↗
PubMedDate not supplied

Declogging small-bore feeding tubes.

Nine substances (Pancrease, Viokase, pork pancreatin, bromelain, papain, cranberry juice, Coca-Cola, chymotrypsin, and distilled water) were tested every half-hour for 4 hr using 900 mm of water pressure to determine their effectiveness in declogging small-bore feeding tubes. At the end of 4 hr the tubes were irrigated with air using a 50-cc syringe. None of the substances tested were effective within 4 hr. Three substances allowed for successful syringe irrigation at the end of 4 hr. They were: chymotrypsin, papain, and distilled water.

Enteral Nutrition↗