Monitoring and Management of Inland Waters

Inland waters are important ecosystems for both their biodiversity and the services they provide to humans [...

Sequencing of β-Peptides by Mass Spectrometry

Seven ?-peptides containing up to 18 β2-, β3- and β2,3-amino acids have been subjected to ESI-tandem mass spectrometry (low-energy fragmentation, positive ions). From the fragment ions formed from the free β-peptides, as well as from the corresponding methyl esters (+14 U) and N-acetyl derivatives (+42 U), the known sequences of β-amino acids could be confirmed unambiguously with the program Sherpa. Thus, the commonly used MS-sequencing procedure for α-peptides can be adopted for β-peptides without modification. However, there are pronounced differences in the fragmentation patterns of the two types of peptides: the β-peptides disclose their relationship to Mannich bases in the mass-spectrometric experiment by the elimination of ammonia from the N-terminus (→ RCH=CH-CO-NH-R') and the occurence of retro-Mannich cleavage (cf. formation of HN=CHR + CH3CO-NH-R' from β-amino-acid residues)

Contaminanti nelle acque lacustri

No abstract availableConfronto von i dati pregressi e analisi temporali della contaminazion

Closely related dermatophyte species produce different patterns of secreted proteins

Dermatophytes are the most common infectious agents responsible for superficial mycosis in humans and animals. Various species in this group of fungi show overlapping characteristics. We investigated the possibility that closely related dermatophyte species with different behaviours secrete distinct proteins when grown in the same culture medium. Protein patterns from culture filtrates of several strains of the same species were very similar. In contrast, secreted protein profiles from various species were different, and so a specific signature could be associated with each of the six analysed species. In particular, protein patterns were useful to distinguish Trichophyton tonsurans from Trichophyton equinum, which cannot be differentiated by ribosomal DNA sequencing. The secreted proteases Sub2, Sub6 and Sub7 of the subtilisin family, as well as Mep3 and Mep4 of the fungalisin family were identified. SUB6, SUB7, MEP3 and MEP4 genes were cloned and sequenced. Although the protein sequence of each protease was highly conserved across species, their level of secretion by the various species was not equivalent. These results suggest that a switch of habitat could be related to a differential expression of genes encoding homologous secreted protein

Thermal regime of a highly regulated Italian river (Ticino River) and implications for aquatic communities

Thermal alteration is one of the adverse effects of flow regulation sharpened in recent years due to climate changes. In this work, we characterize the thermal regime of a highly regulated river located in Northern Italy, which is the emissary of Lake Maggiore. The thermal characteristics of the study reach are influenced by the presence of two dams and by climate warming. In the 15-year monitored period (1999-2013) a significant increase of the mean annual river temperature (i.e., +1.8°C) was indeed recorded. Minimum and maximum water temperatures were detected in correspondence of low-flow periods in February and August, respectively. During August the temperature reached a maximum of 27°C, that is a value below the upper thermal limit of tolerance for most of the aquatic taxa collected in the study area. Moreover, the local presence of seeps and the interaction with groundwater play an important ecological role by guaranteeing patches of cool water to the aquatic communities. Nevertheless, the sensitive early life stages of fish species that spawn in spring/summer may be affected by the high summer temperatures and the expected future climate warming. The wider range of temperatures preferred by alien fish species could result into a competitive disadvantage for autochthonous species. The current minimum flows established by local laws as environmental flows appeared to be able to guarantee an acceptable thermal alteration in morphologically heterogeneous reaches where river/groundwater interaction occurred, at least within the current climatic conditions.</p

The Silkworm as a Source of Natural Antimicrobial Preparations: Efficacy on Various Bacterial Strains

The global spread of multi-resistant pathogens responsible for infections, which cannot be treated with existing drugs such as antibiotics, is of particular concern. Antibiotics are becoming increasingly ineffective and drug resistance is leading to more difficult-to-treat infections; therefore, new bioactive compounds with antimicrobial activity are needed and new alternative sources should be found. Antimicrobial peptides (AMPs) are synthesized by processes typical of the innate immune system and are present in almost all organisms. Insects are extremely resistant to bacterial infections as they can produce a wide range of AMPs, providing an effective first line of defense. The AMPs produced by insects therefore represent a possible source of natural antimicrobial molecules. In this paper, the possibility of using plasma preparations from silkworm (Bombyx mori) larvae as a source of antimicrobials was evaluated. After simple purification steps, insect plasma was analyzed and tested on different Gram-positive and Gram-negative bacterial strains. The results obtained are encouraging as the assays on Escherichia coli and Enterobacter cloacae showed significant decrease in the growth of these Gram-negative bacteria. Similar results were obtained on Gram-positive bacteria, such as Micrococcus luteus and Bacillus subtilis, which showed strong susceptibility to the silkworm AMPs pool. In contrast, Staphylococcus aureus displayed high resistance to Bombyx mori plasma. Finally, the tested plasma formulations were assessed for possible storage not only at 4 \ub0C, but also above room temperature. In conclusion, partially purified plasma from silkworm could be a promising source of AMPs which could be used in formulations for topical applications, without additional and expensive purification steps

Chemical crosslinking and mass spectrometry to elucidate the topology of integral membrane proteins.

Here we made an attempt to obtain partial structural information on the topology of multispan integral membrane proteins of yeast by isolating organellar membranes, removing peripheral membrane proteins at pH 11.5 and introducing chemical crosslinks between vicinal amino acids either using homo- or hetero-bifunctional crosslinkers. Proteins were digested with specific proteases and the products analysed by mass spectrometry. Dedicated software tools were used together with filtering steps optimized to remove false positive crosslinks. In proteins of known structure, crosslinks were found only between loops residing on the same side of the membrane. As may be expected, crosslinks were mainly found in very abundant proteins. Our approach seems to hold to promise to yield low resolution topological information for naturally very abundant or strongly overexpressed proteins with relatively little effort. Here, we report novel XL-MS-based topology data for 17 integral membrane proteins (Akr1p, Fks1p, Gas1p, Ggc1p, Gpt2p, Ifa38p, Ist2p, Lag1p, Pet9p, Pma1p, Por1p, Sct1p, Sec61p, Slc1p, Spf1p, Vph1p, Ybt1p)

Considering mesohabitat scale in ecological impact assessment of sediment flushing

Benthic macroinvertebrates respond to several factors characterizing the physical habitats, as water depth, current and streambed substrate. Thus, anthropogenic disturbances altering these factors may have different effects on benthos, also depending on mesohabitats. These disturbances include sediment flushing operations, commonly carried out to recover reservoir capacity, and investigating their effects at mesohabitat scale could be relevant for an adequate ecological impact assessment of these operations. Here, we compared benthic macroinvertebrate communities sampled before and after a controlled sediment flushing operation in three different mesohabitats (a pool, a riffle and a step-pool) of an Alpine stream. Contrary from expectations, the composition of macroinvertebrate assemblages was not significantly different among mesohabitats. Moreover, the impact of sediment flushing was more significant in terms of density rather than in richness. Two stressor-specific indices were tested, but only one (the Siltation Index for LoTic EcoSystems - SILTES) clearly detected the impact of sediment flushing on the macroinvertebrate community structure. Finally, some differences in the temporal trajectories and recovery times to pre-flushing conditions were observed among mesohabitats, both if the three mesohabitats were considered separately and if all their possible combinations were accounted for. Particularly, riffle was the most sensitive mesohabitat, not fully recovering one year after the sediment disturbance

Proteome-Wide Effect of 17-β-Estradiol and Lipoxin A4 in an Endometriotic Epithelial Cell Line.

Endometriosis affects approximately 10% of women of reproductive age. This chronic, gynecological inflammatory disease results in a decreased quality of life for patients, with the main symptoms including chronic pelvic pain and infertility. The steroid hormone 17-β Estradiol (E2) plays a key role in the pathology. Our previous studies showed that the anti-inflammatory lipid Lipoxin A4 (LXA4) acts as an estrogen receptor-alpha agonist in endometrial epithelial cells, inhibiting certain E2-mediated effects. LXA4 also prevents the progression of endometriosis in a mouse model via anti-proliferative mechanisms and by impacting mediators downstream of ER signaling. The aim of the present study was therefore to examine global proteomic changes evoked by E2 and LXA4 in endometriotic epithelial cells. E2 impacted a greater number of proteins in endometriotic epithelial cells than LXA4. Interestingly, the combination of E2 and LXA4 resulted in a reduced number of regulated proteins, with LXA4 mediating a suppressive effect on E2-mediated signaling. These proteins are involved in diverse pathways of relevance to endometriosis pathology and metabolism, including mRNA translation, growth, proliferation, proteolysis, and immune responses. In summary, this study sheds light on novel pathways involved in endometriosis pathology and further understanding of signaling pathways activated by estrogenic molecules in endometriotic epithelial cells