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Preliminary evaluation of an in vitro test for assessment of excitotoxicity by measurement of early gene (c-fos mRNA) levels

Griffiths, R. (author), Grieve, A. (author), Scollon, J. (author), Scott, M. (author), Williams, C. (author), Meredith, C. (author), TNO Kwaliteit van Leven TNO Bibra (author)

Using primary cultures of mouse cerebellar granule cells as an in vitro model system, it has been demonstrated that different profiles of temporal expression of the c-fos proto-oncogene are observed under non-excitotoxic and excitotoxic conditions. A ratio has been derived previously for the steady- state level of c-fos mRNA after 30 min and 240...

article 2000

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A role for NF-KB-dependent gene transactivation in sunburn

Abeyama, K. (author), Eng, W. (author), Jester, J.V. (author), Vink, A.A. (author), Edelbaum, D. (author), Cockerell, C.J. (author), Bergstresser, P.R. (author), Takashima, A. (author)

Exposure of skin to ultraviolet (UV) radiation is known to induce NF-κB activation, but the functional role for this pathway in UV-induced cutaneous inflammation remains uncertain. In this study, we examined whether experimentally induced sunburn reactions in mice could be prevented by blocking UV-induced, NF-κB-dependent gene transactivation...

article 2000

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Urokinase receptor expression on human microvascular endothelial cells is increased by hypoxia: Implications for capillary-like tube formation in a fibrin matrix

Kroon, M.E. (author), Koolwijk, P. (author), van der Vecht, B. (author), van Hinsbergh, V.W.M. (author), Gaubius Instituut TNO (author)

Hypoxia stimulates angiogenesis, the formation of new blood vessels. This study evaluates the direct effect of hypoxia (1% oxygen) on the angiogenic response of human microvascular endothelial cells (hMVECs) seeded on top of a 3-dimensional fibrin matrix, hMVECs stimulated with fibroblast growth factor-2 (FGF-2) or vascular endothelial growth...

article 2000

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Characterization of a foldase, protein disulfide isomerase A, in the protein secretory pathway of Aspergillus niger

Centraal Instituut voor Voedingsonderzoek TNO TNO Voeding (author), Ngiam, C. (author), Jeenes, D.J. (author), Punt, P.J. (author), van den Hondel, C.A.M.J.J. (author), Archer, D.B. (author)

Protein disulfide isomerase (PDI) is important in assisting the folding and maturation of secretory proteins in eukaryotes. A gene, pdiA, encoding PDIA was previously isolated from Aspergillus niger, and we report its functional characterization here. Functional analysis of PDIA showed that it catalyzes the refolding of denatured and reduced...

article 2000

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The role of biotransformation in dietary (anti)carcinogenesis

van Iersel, M.L.P.S. (author), Verhagen, H. (author), van Bladeren, P.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

The fact that dietary compounds influence the susceptibility of human beings to cancer, is widely accepted. One of the possible mechanisms that is responsible for these (anti)carcinogenic effects is that dietary constituents may modulate biotransformation enzymes, thereby affecting the (anti)carcinogenic potential of other compounds. This...

article 1999

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Selection of Arabidopsis mutants overexpressing genes driven by the promoter of an auxin-inducible glutathione S-transferase gene

van der Kop, D.A.M. (author), Schuyer, M. (author), Pinas, J.E. (author), van der Zaal, B.J. (author), Hooykaas, P.J.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

Transgenic arabidopsis plants were isolated that contained a T-DNA construct in which the promoter of an auxin-inducible glutathione S-transferase (GST) gene from tobacco was fused to the kanamycin resistance (nptII) as well as to the β-glucuronidase (gusA) reporter gene. Subsequently, seeds were treated with EMS to obtain mutants in which both...

article 1999

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Expression of the protective antigen of bacillus anthracis by lactobacillus casei: Towards the development of an oral vaccine against anthrax

Zegers, N.D. (author), Kluter, E. (author), van der Stap, H. (author), van Dura, E. (author), van Dalen, P. (author), Shaw, M. (author), Baillie, L. (author), TNO Preventie en Gezondheid (author)

Bacillus anthracis is the causative organism of the disease anthrax. The ability of the organism to form resistant spores and infect via the aerosol route has led to it being considered as a potential biological warfare agent. The current available human vaccines are far from ideal, they are expensive to produce, require repeated doses and may...

article 1999

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Isolation and characterization of promoter regions from Streptococcus gordonii CH1

Vriesema, A.J.M. (author), Dankert, J. (author), Zaat, S.A.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

We aimed to identify transcription signal sequences from Streptococcus gordonii strain CH1 by random chromosomal cloning. Five genomic fragments from a Sau3A digest, which constitutively activated transcription of a promoterless spectinomycin resistance gene in this strain, were isolated and characterized. Additionally, one promoter fragment was...

article 1999

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Insulin suppresses apolipoprotein(a) synthesis by primary cultures of cynomolgus monkey hepatocytes

Neele, D.M. (author), de Wit, E.C.M. (author), Princen, H.M.G. (author), Gaubius instituut TNO (author)

Raised plasma lipoprotein(a) (lp(a)) concentrations have been reported in patients with Type I (insulin-dependent) diabetes mellitus, which were lowered by insulin therapy. To investigate the biochemical background of these changes, we studied the effect of insulin on apolipoprotein(a) (apo(a)) synthesis and mRNA levels in primary cultures of...

article 1999

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Fibrates suppress fibrinogen gene expression in rodents via activation of the peroxisome proliferator-activated receptor-α

Kockx, M. (author), Gervois, P.P. (author), Poulain, P. (author), Derudas, B. (author), Peters, J.M. (author), Gonzalez, F.J. (author), Princen, H.M.G. (author), Kooistra, T. (author), Staels, B. (author), Gaubius instituut TNO (author)

Plasma fibrinogen levels have been identified as an important risk factor for cardiovascular diseases. Among the few compounds known to lower circulating fibrinogen levels in humans are certain fibrates. We have studied the regulation of fibrinogen gene expression by fibrates in rodents. Treatment of adult male rats with fenofibrate (0.5% [wt/wt...

article 1999

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Lipoprotein cholesterol uptake mediates up-regulation of bile-acid synthesis by increasing cholesterol 7α-hydroxylase but not sterol 27-hydroxylase gene expression in cultured rat hepatocytes

Gaubius instituut TNO (author), Post, S.M. (author), Twisk, J. (author), van der Fits, L. (author), de Wit, E.C.M. (author), Hoekman, M.F.M. (author), Mager, W.H. (author), Princen, H.M.G. (author)

Lipoproteins may supply substrate for the formation of bile acids, and the amount of hepatic cholesterol can regulate bile-acid synthesis and increase cholesterol 7α-hydroxylase expression. However, the effect of lipoprotein cholesterol on sterol 27-hydroxylase expression and the role of different lipoproteins in regulating both enzymes are not...

article 1999

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Fibrate-modulated expression of fibrinogen, plasminogen activator inhibitor-1 and apolipoprotein A-I in cultured cynomolgus monkey hepatocytes. Role of the peroxisome proliferator-activated receptor-α

Kockx, M. (author), Princen, H.M.G. (author), Kooistra, T. (author), Gaubius Instituut TNO (author)

Fibrates are used to lower plasma triglycerides and cholesterol levels in hyperlipidemic patients. In addition, fibrates have been found to alter the plasma concentrations of fibrinogen, plasminogen activator inhibitor-1 (PAI-1) and apolipoprotein A-I (apo A-I). We have investigated the in vitro effects of fibrates on fibrinogen, PAI-1 and apo A...

article 1998

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Effect of steroid hormones and retinoids on the formation of capillary-like tubular structures of human microvascular endothelial cells in fibrin matrices is related to urokinase expression

Lansink, M. (author), Koolwijk, P. (author), van Hinsbergh, V. (author), Kooistra, T. (author), Gaubius Instituut TNO (author)

Angiogenesis, the formation of new capillary blood vessels, is a feature of a variety of pathological processes. To study the effects of a specific group of hormones (all ligands of the steroid/retinoid/thyroid hormone receptor superfamily) on the angiogenic process in humans, we have used a model system in which human microvascular endothelial...

article 1998

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Expression and secretion of defined cutinase variants by Aspergillus awamori

van Gemeren, I.A. (author), Beijersbergen, A. (author), van den Hondel, C.A.M.J.J. (author), Verrips, C.T. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

Several cutinase variants derived by molecular modelling and site- directed mutagenesis of a cutinase gene from Fusarium solani pisi are poorly secreted by Saccharomyces cerevisiae. The majority of these variants are successfully produced by the filamentous fungus Aspergillus awamori. However, the L51S and T179Y mutations caused reductions in...

article 1998

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Expression of heterologous genes in Schizophyllum commune is often hampered by the formation of truncated transcripts

Schuren, F.H.J. (author), Wessels, J.G.H. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

GPD regulatory sequences were used to express a phleomycin resistance gene (Sh ble) in Schizophyllum commune, resulting in high numbers of phleomycin-resistant transformants. Attempts to express heterologous genes coding for hygromycin B phosphotransferase (hph), aminoglycoside phosphotransferase (apt), β-glucuronidase (uidA) and β-galactosidase...

article 1998

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Cloning, sequence analysis, and characterization of the genes involved in isoprimeverose metabolism in Lactobacillus pentosus

Chaillou, S. (author), Lokman, B.C. (author), Leer, R.J. (author), Posthuma, C. (author), Postma, P.W. (author), Pouwels, P.H. (author)

Two genes, xylP and xylQ, from the xylose regulon of Lactobacillus pentosus were cloned and sequenced. Together with the repressor gene of the regulon, xylR, the xylPQ genes form an operon which is inducible by xylose and which is transcribed from a promoter located 145 bp upstream of xylP. A putative xylR binding site (xylO) and a cre-like...

article 1998

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The untranslated leader sequence of the barley lipoxygenase 1 (Lox1) gene confers embryo-specific expression

Rouster, J. (author), van Mechelen, J. (author), Cameron-Mills, V. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

The barley lipoxygenase 1 (Lox1) gene encodes a protein expressed in embryos during grain development and germination and in leaves after methyl-jasmonate (MeJA) treatment. Transient gene expression assays in germinating barley embryos were used to identify cis-regulatory elements involved in the embryo-specific expression of the Lox1 gene....

article 1998

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Lactic acid bacteria as antigen delivery vehicles for oral immunization purposes

Pouwels, P.H. (author), Leer, R.J. (author), Shaw, M. (author), Heijne Den Bak-Glashouwer, M.J. (author), Tielen, F.D. (author), Smit, E. (author), Martinez, B. (author), Jore, J. (author), Conway, P.L. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

In vaccination programmes in which large numbers of subjects are involved, the oral route of administration is more convenient as compared to the more frequently used parenteral route. This is particularly relevant when vaccines are to be applied in less industrialized countries. Lactic acid bacteria in general and strains of Lactobacillus in...

article 1998

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Analysis of the role of the gene bipA, encoding the major endoplasmic reticulum chaperone protein in the secretion of homologous and heterologous proteins in black Aspergilli

Punt, P.J. (author), van Gemeren, I.A. (author), Drint-Kuijvenhoven, J. (author), Hessing, J.G.M. (author), Muijlwijk van - Harteveld, G.M. (author), Beijersbergen, A. (author), Verrips, C.T. (author), van den Hondel, C.A.M.J.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

The function of the endoplasmic-reticulum-localized chaperone binding protein (BiP) in relation to protein secretion in filamentous fungi was studied. It was shown that the overproduction of several homologous and heterologous recombinant proteins by Aspergillus strains induces the expression of bipA, the BiP-encoding gene from Aspergillus niger...

article 1998

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Retinoids increase human apo C-III expression at the transcriptional level via the retinoid X receptor: Contribution to the hypertriglyceridemic action of retinoids

Gaubius Instituut TNO (author), Vu-Dac, N. (author), Gervois, P. (author), Torra, I.P. (author), Fruchart, J.C. (author), Kosykh, V. (author), Kooistra, T. (author), Princen, H.M.G. (author), Dallongeville, J. (author), Staels, B. (author)

Hypertriglyceridemia is a metabolic complication of retinoid therapy. In this study, we analyzed whether retinoids increase the expression of apo C- III, an antagonist of plasma triglyceride catabolism. In men, isotretinoin treatment (80 mg/d; 5 d) resulted in elevated plasma apo C-III, but not apo E concentrations. In human hepatoma HepG2 cells...

article 1998