Aspergillus fumigatus calcineurin interacts with a nucleoside diphosphate kinase

The Ca2+-calcineurin pathway affects virulence and morphogenesis in filamentous fungi. Here, we identified 37 CalA-interacting proteins that interact with the catalytic subunit of calcineurin (CalA) in Aspergillus fumigatus, including the nucleoside diphosphate kinase (SwoH). The in vivo interaction between CalA and SwoH was validated by bimolecular fluorescence complementation. A. fumigatus swoH is an essential gene. Therefore, a temperature-sensitive conditional mutant strain with a point mutation in the active site, SwoH(V83F), was constructed, which demonstrated reduced growth and increased sensitivity to elevated temperatures. The SwoH(V83F) mutation did not cause a loss in virulence in the Galleria mellonella infection model. Taken together these results imply that CalA interacts with SwoH. (C) 2012 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Aspergillus fumigatus mitochondrial electron transport chain mediates oxidative stress homeostasis, hypoxia responses and fungal pathogenesis

We previously observed that hypoxia is an important component of host microenvironments during pulmonary fungal infections. However, mechanisms of fungal growth in these in vivo hypoxic conditions are poorly understood. Here, we report that mitochondrial respiration is active in hypoxia (1% oxygen) and critical for fungal pathogenesis. We generated Aspergillus fumigatus alternative oxidase (aoxA) and cytochrome C (cycA) null mutants and assessed their ability to tolerate hypoxia, macrophage killing and virulence. In contrast to ?aoxA, ?cycA was found to be significantly impaired in conidia germination, growth in normoxia and hypoxia, and displayed attenuated virulence. Intriguingly, loss of cycA results in increased levels of AoxA activity, which results in increased resistance to oxidative stress, macrophage killing and long-term persistence in murine lungs. Thus, our results demonstrate a previously unidentified role for fungal mitochondrial respiration in the pathogenesis of aspergillosis, and lay the foundation for future research into its role in hypoxia signalling and adaptation.

Functional Characterization of an Aspergillus fumigatus Calcium Transporter (PmcA) that Is Essential for Fungal Infection

Aspergillus fumigatus is a primary and opportunistic pathogen, as well as a major allergen, of mammals. The Ca+2-calcineurin pathway affects virulence, morphogenesis and antifungal drug action in A. fumigatus. Here, we investigated three components of the A. fumigatus Ca+2-calcineurin pathway, pmcA,-B, and -C, which encode calcium transporters. We demonstrated that CrzA can directly control the mRNA accumulation of the pmcA-C genes by binding to their promoter regions. CrzA-binding experiments suggested that the 5'-CACAGCCAC-3' and 5'-CCCTGCCCC-3' sequences upstream of pmcA and pmcC genes, respectively, are possible calcineurin-dependent response elements (CDREs)-like consensus motifs. Null mutants were constructed for pmcA and -beta and a conditional mutant for pmcC demonstrating pmcC is an essential gene. The Delta pmcA and Delta pmcB mutants were more sensitive to calcium and resistant to manganese and cyclosporin was able to modulate the sensitivity or resistance of these mutants to these salts, supporting the interaction between calcineurin and the function of these transporters. The pmcA-C genes have decreased mRNA abundance into the alveoli in the Delta calA and Delta crzA mutant strains. However, only the A. fumigatus Delta pmcA was avirulent in the murine model of invasive pulmonary aspergillosis.

Comparative EST analysis provides insights into the basal aquatic fungus Blastocladiella emersonii

Abstract Background Blastocladiella emersonii is an aquatic fungus of the Chytridiomycete class, which is at the base of the fungal phylogenetic tree. In this sense, some ancestral characteristics of fungi and animals or fungi and plants could have been retained in this aquatic fungus and lost in members of late-diverging fungal species. To identify in B. emersonii sequences associated with these ancestral characteristics two approaches were followed: (1) a large-scale comparative analysis between putative unigene sequences (uniseqs) from B. emersonii and three databases constructed ad hoc with fungal proteins, animal proteins and plant unigenes deposited in Genbank, and (2) a pairwise comparison between B. emersonii full-length cDNA sequences and their putative orthologues in the ascomycete Neurospora crassa and the basidiomycete Ustilago maydis. Results Comparative analyses of B. emersonii uniseqs with fungi, animal and plant databases through the two approaches mentioned above produced 166 B. emersonii sequences, which were identified as putatively absent from other fungi or not previously described. Through these approaches we found: (1) possible orthologues of genes previously identified as specific to animals and/or plants, and (2) genes conserved in fungi, but with a large difference in divergence rate in B. emersonii. Among these sequences, we observed cDNAs encoding enzymes from coenzyme B12-dependent propionyl-CoA pathway, a metabolic route not previously described in fungi, and validated their expression in Northern blots. Conclusion Using two different approaches involving comparative sequence analyses, we could identify sequences from the early-diverging fungus B. emersonii previously considered specific to animals or plants, and highly divergent sequences from the same fungus relative to other fungi.

Effect of Byrsonima crassa and Phenolic Constituents on Helicobacter pylori-Induced Neutrophils Oxidative Burst

Byrsonima crassa Niedenzu (Malpighiaceae) is used in Brazilian folk medicine for the treatment of diseases related mainly to gastric ulcers. In a previous study, our group described the gastric protective effect of the methanolic extract from the leaves of B. crassa. The present study was carried out to investigate the effects of methanolic extract and its phenolic compounds on the respiratory burst of neutrophils stimulated by H. pylori using a luminol-based chemiluminescence assay as well as their anti-H. pylori activity. The suppressive activity on oxidative burst of H. pylori-stimulated neutrophils was in the order of methyl gallate > (+)-catechin > methanol extract > quercetin 3-O-alpha-L-arabinopyranoside > quercetin 3-O-beta-D-galactopyranoside > amentoflavone. Methyl gallate, compound that induced the highest suppressive activity with IC50 value of 3.4 mu g/mL, did not show anti-H. pylori activity. B. crassa could be considered as a potential source of natural antioxidant in gastric ulcers by attenuating the effects on the damage to gastric mucosa caused by neutrophil generated reactive oxygen species, even when H. pylori displays its evasion mechanisms.

Tracing latitudinal gradient, river discharge and water masses along the Subtropical South American Coast using benthic Foraminifera assemblages

More than 30% of Buccella peruviana (D'Orbigny), Globocassidulina crassa porrecta (Earland & Heron-Allen), Cibicides mackannai (Galloway & Wissler) and C. refulgens (Montfort) indicate the presence of cold Sub Antarctic Shelf Water in winter, from 33.5 to 38.3 degrees S, deeper than 100 m, in the southern part of the study area. In summer, the abundance of this association decreases to less than 15% around 37.5-38.9 degrees S where two species (Globocassidulina subglobosa (Brady), Uvigerina peregrina (Cushman) take over. G. subglobosa, U. peregrina, and Hanzawaia boueana (D'Orbigny) are found at 27-33 degrees S in both seasons in less than 55 m deep in the northern part, and are linked with warm Subtropical Shelf Water and Tropical Water. Freshwater influence was signalized by high silicate concentration and by the presence of Pseudononion atlanticum (Cushman), Bolivina striatula (Cushman), Buliminella elegantissima (D'Orbigny), Bulimina elongata (D'Orbigny), Elphidium excavatum (Terquem), E. poeyanum (D'Orbigny), Ammobaculites exiguus (Cushman & Bronnimann), Arenoparrella mexicana (Kornfeld), Gaudryina exillis (Cushman & Bronnimann), Textularia earlandi (Parker) and thecamoebians in four sectors of the shelf. The presence of Bulimina marginata (D'Orbigny) between 34.1-32.8 degrees S in the winter and 34.2-32.7 degrees S in the summer indicates that the influence of the Subtropical Shelf Front on the sediment does not change seasonally, otherwise, the presence of Angulogerina angulosa (Williamson) in the winter, only in Mar del Plata (38.9 degrees S), show that Malvinas currents are not influencing the sediment in the summer.

Taxonomic study on the molluscs collected in Marion-Dufresne expedition (MD55) to SE Brazil: Xenophoridae, Cypraeoidea, mitriforms and Terebridae (Caenogastropoda)

The deep-water molluscs collected during the expedition MD55 off SE Brazil have been gradually studied in some previous papers. The present one is focused on samples belonging to caenogastropod taxa Xenophoridae Troschel, 1852, Cypraeoidea Rafinesque, 1815, mitriforms and Terebridae Morch, 1852. Regarding the Xenophoridae, Onustus aquitanus n. sp. is a new species, collected off the littoral of Espirito Santo and Rio de Janeiro, Brazil, 430-637 m depth (continental slope). The main characters of the species include the small size (c. 20 mm), the proportionally wide shell, the white colour, the short peripheral flange, the oblique riblets weakly developed and a brown multispiral protoconch. This appears to be the smallest living species of the family, resembling in this aspect fossil species. In respect to the Cypraeoidea, the following results were obtained: family Cypraeidae Rafinesque, 1815: Erosaria acicularis (Gmelin, 1791) and Luria cinerea (Gmelin, 1791) had the deepest record, respectively 607-620 m and 295-940 m, although the samples were all dead, eroded shells. Family Lamellariidae d'Orbigny, 1841: a total of three lots were collected, provisionally identified as Lamellaria spp. as the samples consist of only vestigial shells; possibly each lot represents a different species. Family Pediculariidae Gray, 1853: a sample of Pedicularia tibia Simone, 2005 was found, expanding the range c. 1000 km southwards, from Ceara to Espirito Santo. Family Ovulidae Fleming, 1822: Pseudosimnia lacrima n. sp., collected off Espirito Santo, 607-620 m depth, is described here and is mainly characterised by its strong biconic outline, small size (c. 7 mm), and a thick peripheral callus. Family Triviidae Troschel, 1863: Cleotrivia antillarum (Schilder, 1922) is recorded for the first time as deep as 620 m, and its distribution expanded from Rio Grande do Norte to Espirito Santo; Dolichupis akangus n. sp. with rounded outline and c. 15 transverse ribs; D. pingius n. sp. with the outer lip expanded posteriorly and c. 10 ribs. In respect to the mitriform neogastropods, the following species are emphasised: family Costellariidae MacDonald, 1860: Vexillum sp., 607-620 m depth; Turricostellaria amphissa n. sp., 295 m depth; T. jukyry n. sp.; T. apyrahi n. sp., both 790-1575 m depth; T. ovir n. sp., 1200 m depth; Nodicostellaria crassa (Simone, 1995), 240-600 m depth, with extension northwards of the range up to Espirito Santo; Austromitra decresca n. sp., 60-105 m depth. Family Mitridae Swainson, 1829: Subcancilla joapyra n. sp., 295 m depth; S. cf. straminea (Adams, 1853), 607-620 m depth. Family Volutomitridae Gray, 1854: Microvoluta corona n. sp., 1500-1575 m depth. Family Mitromorphidae Casey, 1904: Mitromorpha sama n. sp., 607-940 m depth; M. mirim n. sp., 60105 m depth. Regarding the conoidean Terebridae, this paper is a complement of a previous study. It deals with a new species Terebra assu Simone n. sp., from the Abrolhos Bank, 295 m depth, characterised by its narrow outline, yellowish colour, weak sculpture on the last whorls, and a proportionally broad, paucispiral protoconch. A second finding of Terebra alagoensis Lima, Tenorio & Barros, 2007 expands the geographic range from Alagoas to north Espirito Santo. A discussion on the systematics of the "complex Terebra doellojuradoi" in South American coast is also provided, highlighting the improbability of synonymy between T. leptapsis Simone, 1999 and T doellojuradoi Carcelles, 1953. Differences in size, sculpture, spire angulation, aperture, and mainly in protoconch, indicate specific separations. The presently studied terebrids belong to the "complex Terebra doellojuradoi", which encompasses closely related, deep-water, small species, possessing a relatively high degree of endemicity.

Tracing latitudinal gradient, river discharge and water masses along the subtropical South American coast using benthic Foraminifera assemblages

More than 30% of Buccella peruviana (D'Orbigny), Globocassidulina crassa porrecta (Earland & Heron-Allen), Cibicides mackannai (Galloway & Wissler) and C. refulgens (Montfort) indicate the presence of cold Sub Antarctic Shelf Water in winter, from 33.5 to 38.3º S, deeper than 100 m, in the southern part of the study area. In summer, the abundance of this association decreases to less than 15% around 37.5-38.9º S where two species (Globocassidulina subglobosa (Brady), Uvigerina peregrina (Cushman) take over. G. subglobosa, U. peregrina, and Hanzawaia boueana (D'Orbigny) are found at 27-33º S in both seasons in less than 55 m deep in the northern part, and are linked with warm Subtropical Shelf Water and Tropical Water. Freshwater influence was signalized by high silicate concentration and by the presence of Pseudononion atlanticum (Cushman), Bolivina striatula (Cushman), Buliminella elegantissima (D'Orbigny), Bulimina elongata (D'Orbigny), Elphidium excavatum (Terquem), E. poeyanum (D'Orbigny), Ammobaculites exiguus (Cushman & Brönnimann), Arenoparrella mexicana (Kornfeld), Gaudryina exillis (Cushman & Brönnimann), Textularia earlandi (Parker) and thecamoebians in four sectors of the shelf. The presence of Bulimina marginata (D'Orbigny) between 34.1-32.8º S in the winter and 34.2-32.7º S in the summer indicates that the influence of the Subtropical Shelf Front on the sediment does not change seasonally, otherwise, the presence of Angulogerina angulosa (Williamson) in the winter, only in Mar del Plata (38.9º S), show that Malvinas currents are not influencing the sediment in the summer.