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PubMed1994

Timing of fungal invasion using host's ripening hormone as a signal.

In many postharvest fruit diseases, fungi remain latent until the fruit ripens. How the fungus times its infection at ripening of the host is not known. We have found that the volatiles produced by the climacteric tomato, avocado, and banana fruits induce germination and appressorium formation in Colletotrichum gloeosporioides and Colletotrichum musae. Exposure of the spores of these fungi to ethylene, the host's ripening hormone, at </=1 microl/liter, caused germination, branching of the germ tube, and formation of up to six appressoria from a single spore. Propylene, an ethylene analog, but not the hydrocarbon gas methane was able to induce spore germination and multiple appressorium formation. The ethylene effect on the fungi appears to be a plant-like response as it was inhibited by silver ion and 2,5-norbornadiene; the inhibition by the latter could be reversed by higher ethylene concentrations. Ethylene induced germination and appressorium formation in the Colletotrichum sp. penetrating climacteric fruit but not in other Colletotrichum strains. That the ethylene induction of multiple appressorium formation could be relevant to postharvest infection was indicated by the observation that C. gloeosporioides spores formed multiple appressoria on normally ripening tomato that produces ethylene, whereas on transgenic tomato and orange, fruits incapable of producing ethylene, exogenous ethylene was required to induce multiple appressorium formation and lesion formation. These results strongly suggest that these fungi must have coevolved to develop a mechanism to use the host's ripening hormone as a signal to differentiate into multiple infection structure and thus time the infection process.

Journal Article↗
PubMed2002

Identification of differentially expressed transcripts in the human pathogenic fungus Paracoccidioides brasiliensis by differential display.

Paracoccidioides brasiliensis is a dimorphic human pathogenic fungus that is the causal agent of paracoccidioidomycosis, a systemic disease that predominantly affects rural communities in South and Central America. Dimorphism is a common characteristic of systemic human pathogenic fungi. Here we describe the use of differential display (DD) to isolate and identify differentially expressed genes of P. brasiliensis, in the two cell types, yeast (Y) and mycelium (M), as well as at different time intervals during temperature-induced M to Y transition. Using two oligo-deoxythymidine-anchored primers combined with 10 arbitrary ones, we were able to detect the presence of at least 20 differentially transcribed cDNA fragments. Some of these fragments were further analysed by reverse-northern blot and northern blot in order to confirm their differential expression. The M32, M51 and M73 cDNA fragments were specific for the mycelial form of P. brasiliensis. Furthermore, we found two cDNA fragments (M-Y1 and M-Y2) that were upregulated during M-Y transition. This method was efficient and useful in the detection of differentially expressed genes in P. brasiliensis.

DNA, Complementary↗
PubMed2005

The costimulatory molecules CD80, CD86 and OX40L are up-regulated in Aspergillus fumigatus sensitized mice.

Aspergillus fumigatus (Af) is a fungus associated with allergic bronchopulmonary aspergillosis (ABPA) and other allergic diseases. Immune responses in these diseases are due to T and B cell responses. T cell activation requires both Af-specific engagement of the T-cell-receptor as well as interaction of antigen independent costimulatory molecules including CD28-CD80/CD86 and OX40-OX40L interactions. Since these molecules and their interactions have been suggested to have a potential involvement in the pathogenesis of ABPA, we have investigated their role in a model of experimental allergic aspergillosis. BALB/c mice were primed and sensitized with Af allergens, with or without exogenous IL-4. Results showed up-regulation of both CD86 and CD80 molecules on lung B cells from Af-sensitized mice (79% CD86+ and 24% CD80+) and Af/rIL-4-treated mice (90% CD86+ and 24% CD80+) compared to normal controls (36% and 17%, respectively). Lung macrophages in Af-sensitized mice treated or not with IL-4 showed enhanced expression of these molecules. OX40L expression was also up-regulated on lung B cells and macrophages from both Af-sensitized and Af/rIL-4 exposed mice as compared to normal controls. All Af-sensitized animals showed peripheral blood eosinophilia, enhanced total serum IgE and allergen-specific IgG1 antibodies and characteristic lung inflammation. The up-regulation of CD80, CD86 and OX40L molecules on lung B cells and macrophages from Af-allergen exposed mice suggests a major role for these molecules in the amplification and persistence of immunological and inflammatory responses in ABPA.

Allergens↗
PubMed2002

Spore diffusate isolated from some strains of Aspergillus fumigatus inhibits phagocytosis by murine alveolar macrophages.

Aspergillus fumigatus is a ubiquitous fungus that grows in decaying organic matter. It can cause disease in both immunodeficient and immunocompetent patients by using virulence factors to escape the host defenses. Some of these factors, such as a diffusate, released from the spores of A. fumigatus, have previously been described. This diffusate was demonstrated to inhibit oxidative burst and phagocytosis of coated red blood cells. The present study has shown that this substance can inhibit the phagocytosis of A. fumigatus spores by murine alveolar macrophages (MALU) and evaluated the action of this substance. We quantified phagocytosis by MALU cells with and without diffusate and evaluated the inhibition of phagocytosis by testing diffusates from different strains. We conclude that the spore diffusate of some strains of A. fumigatus can reversibly decrease the ability of alveolar macrophages to ingest A. fumigatus spores.

Animals↗
PubMed2001

A novel actin-related protein gene of Colletotrichum gloeosporioides f. sp. malvae shows altered expression corresponding with spore production.

A novel actin-related protein (arp) was found in the plant pathogenic fungus, Colletotrichum gloeosporioides f. sp. malvae (Cgm), which causes anthracnose disease of round-leaved mallow (Malva pusilla). Sequence comparisons showed that this gene, arpA, belongs to the highly divergent 'other arps' category in the current arp classification system. ArpA is most similar to the arp11 gene of Mus musculus but has a unique structure with deletions at the C-terminus similar to that of the arp10 gene of Saccharomyces cerevisiae. A portion of another putative arp gene, arpB, was found immediately downstream of arpA. Expression of arpA was compared to the constitutively expressed Cgm actin gene, actA. In culture, the relative expression of arpA increased when growth conditions favored sporulation. During infection, arpA expression was greatest at the late necrotrophic phase, when sporulation occurred. Arps have been shown to be important in nuclear migration in fungal hyphae, and the expression pattern of arpA indicates that it may have a particular role during sporulation.

Actins↗
PubMed2003

Mycotoxigenic Fusarium and deoxynivalenol production repress chitinase gene expression in the biocontrol agent Trichoderma atroviride P1.

Mycotoxin contamination associated with head blight of wheat and other grains caused by Fusarium culmorum and F. graminearum is a chronic threat to crop, human, and animal health throughout the world. One of the most important toxins in terms of human exposure is deoxynivalenol (DON) (formerly called vomitoxin), an inhibitor of protein synthesis with a broad spectrum of toxigenicity against animals. Certain Fusarium toxins have additional antimicrobial activity, and the phytotoxin fusaric acid has recently been shown to modulate fungus-bacterium interactions that affect plant health (Duffy and Défago, Phytopathology 87:1250-1257, 1997). The potential impact of DON on Fusarium competition with other microorganisms has not been described previously. Any competitive advantage conferred by DON would complicate efforts to control Fusarium during its saprophytic growth on crop residues that are left after harvest and constitute the primary inoculum reservoir for outbreaks in subsequent plantings. We examined the effect of the DON mycotoxin on ecological interactions between pathogenic Fusarium and Trichoderma atroviride strain P1, a competitor fungus with biocontrol activity against a wide range of plant diseases. Expression of the Trichoderma chitinase genes, ech42 and nag1, which contribute to biocontrol activity, was monitored in vitro and on crop residues of two maize cultivars by using goxA reporter gene fusions. We found that DON-producing F. culmorum and F. graminearum strains repressed expression of nag1-gox. DON-negative wild-type Fusarium strains and a DON-negative mutant with an insertional disruption in the tricothecene biosynthetic gene, tri5, had no effect on antagonist gene expression. The role of DON as the principal repressor above other pathogen factors was confirmed. Exposure of Trichoderma to synthetic DON or to a non-DON-producing Fusarium mutant resulted in the same level of nag1-gox repression as the level observed with DON-producing FUSARIUM: DON repression was specific for nag1-gox and had no effect, either positive or negative, on expression of another key chitinase gene, ech42. This is the first demonstration that a target pathogen down-regulates genes in a fungal biocontrol agent, and our results provide evidence that mycotoxins have a novel ecological function as factors in Fusarium competitiveness.

Chitinases↗
PubMed1994

Binding of Paracoccidioides brasiliensis to laminin through surface glycoprotein gp43 leads to enhancement of fungal pathogenesis.

Extracellular matrix protein laminin binds specifically to yeast forms of Paracoccidioides brasiliensis and enhances adhesion of the fungus to the surface of epithelial Madin-Darby canine kidney cells in vitro. Immunoblotting of fungal extracts showed that the gp43 glycoprotein is responsible for adhesion. This was confirmed by binding assays using purified gp43, with a Kd of 3.7 nM. The coating of P. brasiliensis yeast forms with laminin before injection into hamster testicles enhanced the fungus virulence, resulting in a faster and more severe granulomatous disease. These results indicate that interaction of fungi with extracellular matrix elements may constitute a basis for the evolution of fungal infection toward regional spreading and dissemination.

Adhesiveness↗
PubMed1980

Allergic bronchopulmonary aspergillosis complicating cystic fibrosis in childhood.

Allergic bronchopulmonary aspergillosis, known to be associated with cystic fibrosis in older patients, occurred in 7 young atopic children with cystic fibrosis. The diagnosis was suggested by the onset of, or the increase in, asthmatic symptoms accompanied by major chest x-ray changes ranging from total collapse of a lung or lobe to extensive but changing areas of consolidation. Each of the children had a blood eosinophilia, positive type I skin tests to Aspergillus fumigatus, and reversible airways obstruction. Most had a positive type III skin test and circulating precipitins to A. fumigatus, with raised IgE levels which contained specific antibodies to the fungus on radioallergosorbent (RAST) test. None had advanced suppurative chest disease of cystic fibrosis. None was given specific antifungal agents; two received systemic treatment with corticosteroids, the other received additional drugs for their asthma. Two developed total collapse of one lung, one child being only 2 years old. Five have had recurrences of pulmonary shadowing typical of allergic aspergillosis but are not showing significant progression of their cystic fibrosis lung disease. Our experience suggests that there should be an increased awareness of this condition, particularly its association with extensive pulmonary collapse or consolidation in children with cystic fibrosis who are atopic.

Aspergillosis, Allergic Bronchopulmonary↗
PubMed1997

Dermatological conditions in winter in primary health care on Operation Resolute (Bosnia).

The distribution of dermatological conditions has been studied in a total of 1822 consultations with British troops in a primary health care setting on Operation Resolute (Bosnia) between 1 January and 4 March 1996. Approximately one in eight (12%) of the consultations were for skin conditions; eczema was the most common complaint, but, taken as a whole, infections due to virus (excluding warts), fungus and bacteria made up 30%. The overall distribution of diseases was similar to that seen in British general practice.

Adult↗
PubMed2005

Multiple non-ribosomal peptide synthetase genes determine peptaibol synthesis in Trichoderma virens.

Trichoderma virens, an imperfect fungus, is used as a biocontrol agent to suppress plant disease caused by soilborne fungal pathogens. Antimicrobial peptides it produces include peptaibols of 11, 14, and 18 amino acids in length. These peptaibols were previously reported to be synthesized by a non-ribosomal peptide synthetase (NRPS) encoded by the Tex1 gene in strain Tv29-8. The present study examined the Tex1 homolog in a commercially relevant T. virens strain, G20. Although the gene in G20 was 99% identical in DNA sequence to Tex1 in the 15.8 kb compared, gene disruption results indicate that it is only responsible for the production of an 18-mer peptaibol, and not 11-mer and 14-mer peptaibols. Additional NRPS adenylate domains were identified in T. virens and one was found to be part of a 5-module NRPS gene. Although the multimodule gene is not needed for peptaibol synthesis, sequence comparisons suggest that two of the individual adenylate domain clones might be part of a separate peptaibol synthesis NRPS gene. The results indicate a significant diversity of NRPS genes in T. virens that is unexpected from the literature.

Molecular Sequence Data↗
PubMedDate not supplied

Antrolithiasis in the frontal sinus.

A very rare case of sinusitis with antrolithiasis in the left frontal sinus of a 63-year-old male patient is reported. Various conservative treatments had no effect on the decrease of his left frontal pain and of postnasal drip. Neither bacteria nor fungus were detected in the discharge. Computed tomographic scanning revealed several high-dense spots in an isodense shadow in the left frontal sinus. At first, endoscopic sinus surgery (ESS) was employed and a stony mass was detected in the nasofrontal duct and sinus. Patency of the nasofrontal duct was insured and a sticky paste and small masses were removed as well as possible. However, the flow of discharge from the frontal duct continued after surgery. We performed a second operation with extranasal approach and additional stones in the sinus were successfully removed. Most cases of antrolithiasis are caused by a foreign body or caseous sinusitis with fungus. The maxillary sinus is the most common site of disease in antrolithiasis. It is unknown why the present case of antrolithiasis was in the frontal sinus. In such cases of antrolithiasis or cases having pastelike contents in the frontal sinuses, we conclude that ESS may be an unsuccessful treatment and a classical surgical approach may be required.

Endoscopy↗