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Development of a Fusarium graminearum Affymetrix GeneChip for profiling fungal gene expression in vitro and in planta.

Recently the genome sequences of several filamentous fungi have become available, providing the opportunity for large-scale functional analysis including genome-wide expression analysis. We report the design and validation of the first Affymetrix GeneChip microarray based on the entire genome of a filamentous fungus, the ascomycetous plant pathogen Fusarium graminearum. To maximize the likelihood of representing all putative genes (approximately 14,000) on the array, two distinct sets of automatically predicted gene calls were used and integrated into the online F. graminearum Genome DataBase. From these gene sets, a subset of calls was manually annotated and a non-redundant extract of all calls together with additional EST sequences and controls were submitted for GeneChip design. Experiments were conducted to test the performance of the F. graminearum GeneChip. Hybridization experiments using genomic DNA demonstrated the usefulness of the array for experimentation with F. graminearum and at least four additional pathogenic Fusarium species. Differential transcript accumulation was detected using the F. graminearum GeneChip with treatments derived from the fungus grown in culture under three nutritional regimes and in comparison with fungal growth in infected barley. The ability to detect fungal genes in planta is surprisingly sensitive even without efforts to enrich for fungal transcripts. The Plant Expression Database (PLEXdb, http://www.plexdb.org) will be used as a public repository for raw and normalized expression data from the F. graminearum GeneChip. The F. graminearum GeneChip will help to accelerate exploration of the pathogen-host pathways that may involve interactions between pathogenicity genes in the fungus and disease response in the plant.

Computational Biology↗

Inhibition of fungal growth by Pseudomonas aeruginosa and Pseudomonas cepacia isolated from patients with cystic fibrosis.

This study was undertaken because of the infrequency of infections due to Candida species in patients with cystic fibrosis despite their extensive treatment with broad-spectrum antibiotics. In vitro susceptibility studies revealed significant inhibition of 11 strains of fungi known to infect human beings by 10 strains of Pseudomonas aeruginosa and nine strains of Pseudomonas cepacia isolated from the sputum of patients with cystic fibrosis. The fungi were Candida krusei, Candida keyfr, Candida guillermondii, Candida tropicalis, Candida lusitaniae, Candida parapsilosis, Candida pseudotropicalis, Candida albicans, Torulopsis glabrata, Saccharomyces cerevisiae and Aspergillus fumigatus. Inhibition of fungal growth by Escherichia coli (NCTC 10418), Staphylococcus aureus (NCTC 6571) and Haemophilus influenzae (NCTC 11931) could not be demonstrated. The continued presence in the sputum of patients with cystic fibrosis of strains of P. aeruginosa and P. cepacia, which produce antifungal substances, may inhibit growth of Candida species and so prevent overt Candida infections. A. fumigatus would seem to be the most important fungus causing disease in patients with cystic fibrosis. It is therefore interesting to note that this was the most resistant of all the fungi tested for inhibition by P. aeruginosa and P. cepacia.

Antibiosis↗

Mycology.

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Humans↗

Antemortem diagnosis of systemic aspergillosis: ten-year review and report of a case.

A review of the medical literature over the last ten years indicates the rarity of antemortem diagnosis and treatment of systemic aspergillosis despite its rapid increase in incidence in the United States. We report a case of systemic aspergillosis caused by Aspergillus flavus which was isolated from the blood of an immunosuppressed patient with chronic obstructive pulmonary disease. The fungus grew rapidly on brain-heart infusion medium and Sabouraud dextrose agar cultures. Autopsy examination of tissue and subsequent cultures confirmed the antemortem diagnosis.

Aspergillosis↗