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  • Journal article
    Scheid P, Griesenbach U, Davies J, Siddique Z, Gruenert DC, Geddes DM, Alton EWFWet al., 1998,

    Interleukin-8 production by airway epithelium and the effect of liposome-mediated IκB gene transfer

    , Thorax, Vol: 53, ISSN: 0040-6376

    Interleukin-8 (IL-8) is a major chemoattractant for neutrophils, present in the CF lung in large numbers. We measured basal and stimulated IL-8 secretion from CF and non CF airway epithelial cell lines (CFTE and 16HBE cells, respectively) as well as from Cos7 cells in a controlled in vitro environment. Basal IL-8 secretion was significantly higher in CFTE cells (8110 ± 2286 pg/mg protein) than in 16HBE (1113 ± 123 pg/mg protein) and Cos7 (497 ± 138 pg/mg protein) [n=6, p<0.05]. Following TNF-α stimulation IL-8 secretion increased 8 ± 0.9 fold in CFTE, 2 ± 0.4 fold in 16HBE and 2.7 ± 0.2 fold in Cos7 cells. Preliminary data in freshly obtained human nasal cells also indicate an increased basal secretion of IL-8 (non CF: 62 ± 10 pg/ml n=16, CF: 165 ± 37 pg/ml n=12, p<0.05). The response to addition of Pseudomonas aeruginosa to these cells as measured by IL-8 secretion was also increased in CF (3279 ± 656 pg/ml n=10, p<0.05) when compared to non CF samples (817 ± 143 pg/ml n=8). Thus it appears that the IL-8 response is disproportionately greater in CF cells. It is likely that new treatment aimed at reducing IL-8 levels will be beneficial in CF. Nuclear factor kappa B (NFκB) is a transcription factor that regulates expression of inflammatory mediators including IL-8. NFκB is kept in an inactive state by association with a specific inhibitor (IκB). We investigated the effects of liposome-mediated overexpression of IκB in CFTE and Cos7 cells. Despite successful transfection, as confirmed by RT-PCR in both cell types, we did not detect an alteration of basal or TNF-α stimulated IL-8 secretion. This is most likely related to low levels of gene expression. In conclusion, we have shown that CF cells lines as well as CF primary cells produce increased levels of secreted IL-8 in the basal as well as in a stimulated state. Reduction of IL-8 secretion is likely to b
  • Journal article
    Palmer LJ, Daniels SE, Rye PJ, Gibson NA, Tay GK, Cookson WO, Goldblatt J, Burton PR, LeSöuef PNet al., 1998,

    Linkage of chromosome 5q and 11q gene markers to asthma-associated quantitative traits in Australian children.

    , Am J Respir Crit Care Med, Vol: 158, Pages: 1825-1830, ISSN: 1073-449X

    Asthma is a genetically complex disease, and the investigation of putative linkages to candidate loci in independent populations is an important part of the gene discovery process. This study investigated the linkage of microsatellite markers in the 5q and 11q regions to asthma-associated quantitative traits in 121 Australian Caucasian nuclear families. The families were recruited on the basis of a child proband: a cohort of 95 randomly recruited families of unselected probands (n = 442 subjects) and a cohort of 26 families of probands selected on the basis of severe symptomatic asthma (n = 134 subjects). The quantitative traits assessed included serum levels of total IgE and specific IgE to house dust mite and mixed grass, blood eosinophil counts, and the dose-response slope (DRS) of FEV1 to histamine provocation. Multipoint linkage analysis using Haseman-Elston sib-pair methods provided evidence of significant linkage between the chromosome 5q markers and loge total serum IgE levels, specific serum IgE levels, and loge blood eosinophil counts. The chromosome 11q markers showed evidence of significant linkage to specific serum IgE levels. Neither region demonstrated significant linkage to the loge DRS to histamine. Phenotypes were residualized for age and sex. These data are consistent with the existence of loci regulating asthma-associated quantitative traits in both the 5q31-33 and 11q13 chromosomal regions.
  • Journal article
    Caddick MX, Arst HN, 1998,

    Deletion of the 389 N-terminal residues of the transcriptional activator AREA does not result in nitrogen metabolite derepression in <i>Aspergillus nidulans</i>

    , JOURNAL OF BACTERIOLOGY, Vol: 180, Pages: 5762-5764, ISSN: 0021-9193
  • Journal article
    Denison SH, Negrete-Urtasun S, Mingot JR, Tilburn J, Mayer WA, Goel A, Espeso EA, Peñalva MA, Arst Net al., 1998,

    Putative membrane components of signal transduction pathways for ambient pH regulation in <i>Aspergillus</i> and meiosis in <i>Saccharomyces</i> are homologous

    , MOLECULAR MICROBIOLOGY, Vol: 30, Pages: 259-264, ISSN: 0950-382X
  • Journal article
    Hamada W, Spanu PD, 1998,

    Co-suppression of the hydrophobin gene <i>HCf-1</i> is correlated with antisense RNA biosynthesis in <i>Cladosporium fulvum</i>

    , MOLECULAR AND GENERAL GENETICS, Vol: 259, Pages: 630-638, ISSN: 0026-8925
  • Journal article
    Byrne B, Klahn S, Taylor PL, Eidne KAet al., 1998,

    Functional analysis of GnRH receptor ligand binding using biotinylated GnRH derivatives

    , MOLECULAR AND CELLULAR ENDOCRINOLOGY, Vol: 144, Pages: 11-19, ISSN: 0303-7207
  • Journal article
    Wilson RA, Arst HN, 1998,

    Mutational analysis of AREA, a transcriptional activator mediating nitrogen metabolite repression in <i>Aspergillus nidulans</i> and a member of the "streetwise" GATA family of transcription factors

    , MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS, Vol: 62, Pages: 586-+, ISSN: 1092-2172
  • Journal article
    Wilson RA, Arst HN, 1998,

    Mutational analysis of AREA, a transcriptional activator mediating nitrogen metabolite repression in Aspergillus nidulans and a member of the "streetwise" GATA family of transcription factors.

    , Microbiol Mol Biol Rev, Vol: 62, Pages: 586-596, ISSN: 1092-2172

    The transcriptional activator AREA is a member of the GATA family of transcription factors and mediates nitrogen metabolite repression in the fungus Aspergillus nidulans. The nutritional versatility of A. nidulans and its amenability to classical and reverse genetic manipulations make the AREA DNA binding domain (DBD) a useful model for analyzing GATA family DBDs, particularly as structures of two AREA-DNA complexes have been determined. The 109 extant mutant forms of the AREA DBD surveyed here constitute one of the highest totals of eukaryotic transcription factor DBD mutants, are discussed in light of the roles of individual residues, and are compared to corresponding mutant sequence changes in other fungal GATA factor DBDs. Other topics include delineation of the DBD using both homology and mutational truncation, use of frameshift reversion to detect regions of tolerance to mutational change, the finding that duplication of the DBD can apparently enhance AREA function, and use of the AREA system to analyze a vertebrate GATA factor DBD. Some major points to emerge from work on the AREA DBD are (i) tolerance to sequence change (with retention of function) is surprisingly great, (ii) mutational changes in a transcription factor can have widely differing, even opposing, effects on expression of different structural genes so that monitoring expression of one or even several structural genes can be insufficient and possibly misleading, and (iii) a mutational change altering local hydrophobic packing and DNA binding target specificity can markedly influence the behavior of mutational changes elsewhere in the DBD.
  • Journal article
    Moffatt MF, Cookson WO, 1998,

    Gene identification in asthma and allergy.

    , Int Arch Allergy Immunol, Vol: 116, Pages: 247-252, ISSN: 1018-2438

    Asthma and atopy are due to the interaction between genetic and environmental factors. The genetic basis of asthma and atopy is becoming more certain, with the promise of improvements in the diagnosis and treatment of these disorders. A combination of candidate gene and positional cloning studies has already identified several genes which influence allergic disease. These genes predispose in general to atopy, or influence the specific IgE response to individual allergens, or enhance inflammation independently of atopic mechanisms. In addition to these known factors, the chromosomal localisation of other putative genes is becoming clear.
  • Journal article
    Karpovich-Tate N, Spanu P, Dewey FM, 1998,

    Use of monoclonal antibodies to determine biomass of <i>Cladosporium fulvum</i> in infected tomato leaves

    , MOLECULAR PLANT-MICROBE INTERACTIONS, Vol: 11, Pages: 710-716, ISSN: 0894-0282

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