Maria Guglielminetti

Lung injury and degradation of extracellular matrix components by Aspergillus fumigatus serine proteinase

Aspergillus fumigatus produces a variety of extracellular proteinases that are believed to be virulence factors towards Aspergillus-related lung disease. Among Aspergillus proteinases, the serine proteinase is thought to play a major virulent role because of its widespread production. Nevertheless, evidence of direct pulmonary injury caused by the A. fumigatus serine proteinase is still lacking. The purpose of our work was: (1) to provide evidence for a pivotal role of A. fumigatus serine proteinase in producing lung injury in an animal model, and (2) to investigate the broadness of the substrate specificity of the proteinase towards extracellular matrix components. To achieve this aim, the proteinase from an A. fumigatus strain isolated from human airways was purified by a four-step procedure, including cation exchange and hydrophobic interaction. High-performance capillary electrophoresis, SDS-PAGE, determination of K(m) towards synthetic substrates, and inhibitory studies were used to further characterize the A. fumigatus serine proteinase. With reference to extracellular matrix components, the A. fumigatus serine proteinase was shown to degrade human lung elastin at a higher rate than an equimolar amount of human neutrophil elastase. Human lung collagen, type I and type III collagens, as well as fibronectin, were quickly digested by the A. fumigatus serine proteinase. Finally, mice intratracheally injected with the proteinase showed a significant degree of lower respiratory tract destruction. We conclude that the A. fumigatus serine proteinase is capable per se of hydrolyzing the major structural barriers of the lung.

Mycological and ultrastructural studies to evaluate biodeterioration of mural paintings. Detection of fungi and mites in Frescos of the monastery of St Damian in Assisi

Abstract The role of fungi in the biodeterioration of frescos has been investigated by microbiological and ultrastructural techniques. With this aim, the mycoflora present on samples taken from deteriorated indoor wall paintings (frescos) in the Monastery of St Damian in Assisi was isolated and identified. The results showed that the fungal colonization of the two paintings ‘Crocifisso con Francesco giovane’ and the ‘Mensa di S. Chiara’, located inside St Clares Refectory, was mainly due to Cladosporium, Aspergillus, Alternaria, Penicillium and Fusarium genera. Although considered not uncommon in frescos, their direct involvement in enzymatic degradation of paints was not confirmed. Interestingly, the presence of mites in proximity to actively growing fungal mycelium, as revealed by scanning electron microscopy, although considered an example of parasitic nutritional relation, could also suggest a metabolic interaction resulting in the synthesis of biodeteriogenic substances.

Biodiversity of rock, beach and water fungi in Italy

Abstract The fungal biodiversity in its overall is mostly still unknown and the ecological role of these organisms, particularly in some border ecosystems, is often underestimated. This study aims to give both an overview of the state of the art and to present new data on the mycodiversity in some peculiar environments as rocks, beach sand, and water in Italy. Particularly, rock fungi are here reported from high mountain peaks, sea cliffs, and monuments; sand associated fungi from beach ecosystems in Puglia and Ligurian coasts; marine fungi associated with the endemic seagrass of the Mediterranean Posidonia oceanica L.; aquatic hyphomycetes (Ingoldian fungi) from both streams in the Ticino Natural Park and lentic water in Lago Maggiore; fungi from the water distribution system in Turin. Ecological and evolutive considerations are put forward.

High spots for diversity of soil and litter microfungi in Italy

Abstract Soil fungal diversity plays a fundamental role in delivering key ecosystem goods and services. This article assesses diversity of saprobic soil and litter microfungi, as taxonomical and functional components which affect above- and below-ground relationships within Alpine and Mediterranean regions of Italy. We highlighted biodiversity high spots focusing on four research topics that have been developed over time and are currently in progress in Italy. Preliminary quantitative data concerning soil microfungi in the Raethian Alps showed a strong reduction of Colony Forming Unit (CFU)s with altitude. Keratinophilic microfungi in natural and anthropogenic environments were widespread among filamentous fungi and 121 species have been isolated in Italy since 1960. Heat stimulated microfungi in Mediterranean region soils showed high values both in abundance and species density even two years after the experimental fire, with Neosartorya spp. playing a pivotal role. The diversity of microfungi of Quercus ilex (150 species), in the Mediterranean region, higher than that in leaf litter of other species, was explained mainly by different forms of growth and the phytoclimatic characters of the areas under study.

Respiratory syndrome very similar to extrinsic allergic alveolitis due to Penicillium verrucosum in workers in a cheese factory.

A respiratory syndrome very similar to extrinsic allergic alveolitis due to Penicillium verrucosum was recognized in 4 workers employed in a Gorgonzola cheese factory. A mycogen allergy to P. verrucosum, used as starter in the production, was demonstrated by positive sputum culture and detection of specific antibodies in the blood. Intense and prolonged exposure to inhalation of fungal spores could have lead to the development of this allergic response. The fact that 2 of the subjects are siblings seems to indicate host susceptibility or immunological constitution in the pathogenesis of the respiratory allergy.

Chemical Composition and Antimicrobial Activity of the Volatile Fractions from Leaves and Flowers of the Wild Iraqi Kurdish Plant Prangos peucedanifolia Fenzl

The volatile fractions isolated from Prangos peucedanifolia Fenzl leaves and flowers were investigated for their phytochemical composition and biological properties. Flower and leaf hydrodistillation afforded 3.14 and 0.49 g of yellowish oils in 1.25 and 0.41% yields, respectively, from dry vegetable materials. According to the GC‐FID and GC/MS analyses, 36 (99.35% of the total oil composition) and 26 compounds (89.12%) were identified in the two oils, respectively. The major constituents in the flower volatile fraction were β‐pinene (35.58%), α‐pinene (22.13%), and β‐phellandrene (12.54%), while m‐cresol (50.38%) was the main constituent of the leaf volatile fraction. The antimicrobial activity was evaluated against several bacterial and fungal strains, on the basis of the minimum inhibitory concentration (MIC) by the micro‐ and macrodilution methods. The two volatile fractions showed moderate antifungal and antibacterial activities, especially against Trichophyton rubrum (MIC of 2×103 μg/ml), Streptococcus mutans, Streptococcus pyogenes, and Staphylococcus aureus (MIC≤1.9×103 μg/ml for all).

Chemical composition, enantiomeric analysis, AEDA sensorial evaluation and antifungal activity of the essential oil from the Ecuadorian plant Lepechinia mutica Benth (Lamiaceae)

This study describes the GC‐FID, GC/MS, GC‐O, and enantioselective GC analysis of the essential oil hydrodistilled from leaves of Lepechinica mutica (Lamiaceae), collected in Ecuador. GC‐FID and GC/MS analyses allowed the characterization and quantification of 79 components, representing 97.3% of the total sample. Sesquiterpene hydrocarbons (38.50%) and monoterpene hydrocarbons (30.59%) were found to be the most abundant volatiles, while oxygenated sesquiterpenes (16.20%) and oxygenated monoterpenes (2.10%) were the minor components. In order to better characterize the oil aroma, the most important odorants, from the sensorial point of view, were identified by Aroma Extract Dilution Analysis (AEDA) GC‐O. They were α‐Pinene, β‐Phellandrene, and Dauca‐5,8‐diene, exhibiting the characteristic woody, herbaceus, and earthy odors, respectively. Enantioselective GC analysis of L. mutica essential oil revealed the presence of twelve couples and two enantiomerically pure chiral monoterpenoids. Their enantiomeric excesses were from a few percent units to 100%. Moreover, the essential oil exhibited moderate in vitro activity against five fungal strains, being especially effective against M. canis, which is a severe zoophilic dermatophyte causal agent of pet and human infections.

Phytochemical researches and antimicrobial activity of Clinopodium nubigenum Kunth (Kuntze) raw extracts

The essential oil of the species Clinopodium nubigenum (Kunth) Kuntze, Lamiaceae, was analyzed by GC-MS and GC-FID, taking into account the more recent literature. Among the seventy compounds identified, the majority are oxygenated monoterpenoids. The essential oil, tested for antimicrobial activity, resulted effective in vitro against Candida albicans. From the aqueous MeOH extract of the aerial parts of the plant two nonvolatile compounds, named schizonepetoside A and schizonepedoside C, have been isolated. They are rare glycosyl terpenoids, which were previously isolated from only one plant, but never found before in the genus Clinopodium.

Dietary Supplementation of Hericium erinaceus Increases Mossy Fiber-CA3 Hippocampal Neurotransmission and Recognition Memory in Wild-Type Mice

Hericium erinaceus (Bull.) Pers. is a medicinal mushroom capable of inducing a large number of modulatory effects on human physiology ranging from the strengthening of the immune system to the improvement of cognitive functions. In mice, dietary supplementation with H. erinaceus prevents the impairment of spatial short-term and visual recognition memory in an Alzheimer model. Intriguingly other neurobiological effects have recently been reported like the effect on neurite outgrowth and differentiation in PC12 cells. Until now no investigations have been conducted to assess the impact of this dietary supplementation on brain function in healthy subjects. Therefore, we have faced the problem by considering the effect on cognitive skills and on hippocampal neurotransmission in wild-type mice. In wild-type mice the oral supplementation with H. erinaceus induces, in behaviour test, a significant improvement in the recognition memory and, in hippocampal slices, an increase in spontaneous and evoked excitatory synaptic current in mossy fiber-CA3 synapse. In conclusion, we have produced a series of findings in support of the concept that H. erinaceus induces a boost effect onto neuronal functions also in nonpathological conditions.

Evaluation of the antigen from chlamydospores of Candida albicans in the serodiagnosis of candidiasis

Antigens have been prepared from the chlamydospores and blastospores of Candida albicans and their precipitin patterns were analysed by two-dimensional immunoelectrophoresis using specific antisera.The two antigens were used in routine serological tests of patients suffering from candidiasis. On double-diffusion tests for the detection of circulating antibodies of Candida albicans, the antigen from chlamydospores displays precipitin lines that differ in number and intensity from those obtained with the antigen from blastospores. The results are briefly discussed in the framework of C. albicans antigen standardization.