Chitin Deacetylases: Properties and Applications

Chitin deacetylases, occurring in marine bacteria, several fungi and a few insects, catalyze the deacetylation of chitin, a structural biopolymer found in countless forms of marine life, fungal cell and spore walls as well as insect cuticle and peritrophic matrices. The deacetylases recognize a sequence of four GlcNAc units in the substrate, one of which undergoes deacetylation: the resulting chitosan has a more regular deacetylation pattern than a chitosan treated with hot NaOH. Nevertheless plain chitin is a poor substrate, but glycolated, reprecipitated or depolymerized chitins are good ones. The marine Vibrio sp. colonize the chitin particles and decompose the chitin thanks to the concerted action of chitinases and deacetylases, otherwise they could not tolerate chitosan, a recognized antibacterial biopolymer. In fact, chitosan is used to prevent infections in fishes and crustaceans. Considering that chitin deacetylases play very important roles in the biological attack and defense systems, they may find applications for the biological control of fungal plant pathogens or insect pests in agriculture and for the biocontrol of opportunistic fungal human pathogens.     

Control of brown spot of pear by reducing the overwintering inoculum through sanitation

Stemphylium vesicarium, the causal agent of brown spot of pear, overwinters in the leaf residues of pear and herbaceous plants of the orchard floor. Pseudothecia of the teleomorph, Pleospora allii, are formed on these residues where they produce ascospores. New methods were tested aimed at reducing this overwintering inoculum and increasing the efficacy of control of brown spot of pear. Sanitation methods were evaluated in nine trials in Girona (Spain) and Ferrara (Italy) over a 4-year period. The sanitation methods were leaf litter removal in December to February, and application of biological control agents (commercial formulates of Trichoderma spp.) to the orchard ground cover from February to May. Fungicides were also applied to the trees during the pear-growing season, scheduled according to the BSPcast model. The different methods were tested as stand-alone applications or in combination. All methods consistently reduced the disease incidence at harvest on fruit with an efficacy between 30 to 60% for leaf litter removal and more than 60% for the combination of leaf litter removal and biological control. Efficacy of sanitation alone (leaf litter removal and biological control) in reducing the brown spot level on fruit was similar in most of the trials to the efficacy obtained when fungicides were applied alone. However, integration of sanitation methods and fungicides did not improve the efficacy of disease control over the level provided by fungicides alone.     

Combining sanitation and disease modelling for control of grapevine powdery mildew

Chasmothecia of Erysiphe necator form in one season, survive winter and discharge ascospores that cause primary infections and trigger powdery mildew epidemics in the next season. A strategy for powdery mildew control was developed based on (i) the reduction in overwintering chasmothecia and on (ii) spring fungicide applications to control ascosporic infections timed based on estimate risk (two to five sprays per season). Several fungicides, the hyperparasite Ampelomyces quisqualis, and a mineral oil product were first tested as separate applications in a greenhouse and in vineyards. In the greenhouse, A. quisqualis suppressed chasmothecia formation by 41 %; fungicides and mineral oil suppressed chasmothecia formation by 63 % and ascospore viability by 71 %. In vineyards, application of boscalid + kresoxim-methyl or meptyldinocap once after harvest, as well as application of A. quisqualis pre- and post-harvest, delayed disease onset and epidemic development in the following season by 1 to 3 weeks and lowered disease severity (up to the pea-sized berry stage) by 56 to 63 %. Risk-based applications of sulphur and of synthetic fungicides provided the same control as the grower spray program but required fewer applications (average reduction of 47 %). Sanitation strategies were then tested by combining products and application times (late-season, and/or pre-bud break, and/or spring). Adequate disease control with a reduced number of sprays was achieved with the following combination: two applications of A. quisqualis (pre- and post-harvest), one application of mineral oil before bud break, and model-based applications of sulphur fungicides between bud break and fruit set.     

Ellagic Acid and Polyhydroxylated Urolithins Are Potent Catalytic Inhibitors of Human Topoisomerase II: An in Vitro Study

Ellagic acid (EA), a natural polyphenol abundant in fruits and common in our diet, is under intense investigation for its chemopreventive activity resulting from multiple effects. EA inhibits topoisomerase II, but the effects on the human enzyme of urolithins, its monolactone metabolites, are not known. Therefore, the action of several synthetic urolithins toward topoisomerases II was evaluated, showing that polyhydroxylated urolithins, EA, and EA-related compounds are potent inhibitors of the α and β isoforms of human topoisomerase II at submicromolar concentrations. Competition tests demonstrate a dose-dependent relationship between ATP and the inhibition of the enzyme. Docking experiments show that the active compounds bind the ATP pocket of the human enzyme, thus supporting the hypothesis that EA and polyhydroxylated urolithins act as ATP-competitive inhibitors of human topoisomerase II.     

Long-term evaluation of yield components of young olive trees during the onset of fruit production under different irrigation regimes

A four-year study was conducted on young Olea europaea L. trees to investigate the effect of deficit irrigation starting from the onset of fruit production. Subsurface drip irrigation was used to supply 100% (FI), 46–52% (DI), or 2–6% (SI) of tree water needs. Tree growth was reduced by deficit irrigation, whereas, return bloom was not. Per tree fruit yield of DI trees was 68% that of FI, but fruit yield efficiency based on tree size was similar between treatments. Fruit set and the number of fruits of FI trees were similar to those of DI trees and significantly higher than in SI trees. No significant differences in fruit fresh weight were found between FI and DI. The oil yield and oil yield efficiency of the DI treatment were 82 and 110% that of FI trees, respectively. A level of about 50% deficit proved sustainable to irrigate trees for oil production.     

The Italian veterinary medicine admission test: analysis of student intake in the years 2007, 2008, and 2009, and of the test's relationship with students' academic careers

The present paper analyzes the admission test administered to candidates to the veterinary medicine program in Italy for the academic years 2007-2008, 2008-2009, and 2009-2010 nationwide as well as the University of Pisa student intake from 2001-2002 through 2009-2010, comparing the relationship between the admission test and students' academic careers at Pisa. This paper finds that the Italian system of a locally enforced fixed intake number does not select the best possible candidates for admission because (1) there are significant variations in the candidates' preparation among the different locations where the test is held, (2) the subjects tested are not equally selective in identifying the best candidates, and (3) there is a very strong relationship between candidates' performance on the admission test and the subsequent academic career of the admitted candidates. In its findings, this study in part contradicts what is commonly believed by the Italian veterinary medicine community, and, as a result, it is extremely important that care is taken in the decision-making process-in the process, that is, of identifying a fixed intake number and of selecting the subjects to be tested on the admission test.     

Drying and wetting of Mediterranean soils stimulates decomposition and carbon dioxide emission: the "Birch effect"

Observations on the net carbon exchange of forests in the European Mediterranean region, measured recently by the eddy covariance method, have revived interest in a phenomenon first characterized on agricultural and forest soils in East Africa in the 1950s and 1960s by H. F. Birch and now often referred to as the "Birch effect." When soils become dry during summer because of lack of rain, as is common in regions with Mediterranean climate, or are dried in the laboratory in controlled conditions, and are then rewetted by precipitation or irrigation, there is a burst of decomposition, mineralization and release of inorganic nitrogen and CO(2). In forests in Mediterranean climates in southern Europe, this effect has been observed with eddy covariance techniques and soil respiration chambers at the stand and small plot scales, respectively. Following the early work of Birch, laboratory incubations of soils at controlled temperatures and water contents have been used to characterize CO(2) release following the rewetting of dry soils. A simple empirical model based on laboratory incubations demonstrates that the amount of carbon mineralized over one year can be predicted from soil temperature and precipitation regime, provided that carbon lost as CO(2) is taken into account. We show that the amount of carbon returned to the atmosphere following soil rewetting can reduce significantly the annual net carbon gain by Mediterranean forests.     

Total Phenolic Levels,In Vitro Antioxidant Properties,and Fatty Acid Profile of Two Microalgae,Tetraselmis marina Strain IMA043 and Naviculoid Diatom Strain IMA053, Isolated from the North Adriatic Sea

This work studied the potential biotechnological applications of a naviculoid diatom (IMA053) and a green microalga (Tetraselmis marina IMA043) isolated from the North Adriatic Sea. Water, methanol, and dichloromethane (DCM) extracts were prepared from microalgae biomass and evaluated for total phenolic content (TPC) and in vitro antioxidant properties. Biomass was profiled for fatty acid methyl esters (FAME) composition. The DCM extracts had the highest levels of total phenolics, with values of 40.58 and 86.14 mg GAE/g dry weight (DW in IMA053 and IMA043, respectively). The DCM extracts had a higher radical scavenging activity (RSA) than the water and methanol ones, especially those from IMA043, with RSAs of 99.65% toward 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt (ABTS) at 10 mg/mL, and of 103.43% against 2,2-diphenyl-1-picrylhydrazyl (DPPH) at 5 mg/mL. The DCM extract of IMA053 displayed relevant copper chelating properties (67.48% at 10 mg/mL), while the highest iron chelating activity was observed in the water extract of the same species (92.05% at 10 mg/mL). Both strains presented a high proportion of saturated (SFA) and monounsaturated (MUFA) fatty acids. The results suggested that these microalgae could be further explored as sources of natural antioxidants for the pharmaceutical and food industry and as feedstock for biofuel production.     

Relationships between biological and physico-chemical soil characteristics in a mediterranean agricultural area

Fourteen agricultural soils from various areas of Tuscany were characterized by a range of measurements indicative of soil biological activity. The objective of our research was to identify soil parameters suitable as indicators for evaluating their quality. In general, enzyme activities were found to vary widely, with the highest activity for each enzyme being distributed among only five of the 14 soils studied. The narrowest range (14-fold) in enzyme activities for the various soils was observed for catalase and the widest range (577-fold) for g -glucosidase. Biomass C and, among the measured enzyme activities, amylase, were well correlated with total organic carbon, total N, cation and anion exchange capacity. Positive correlations were found between the maximum water holding capacity and dehydrogenase, amylase, biomass C, FDA hydrolytic activity, the biological index of fertility and the enzyme activity number, so showing that soil moisture may play an important role in affecting soil biological characteristics. No significant correlations were observed among the soil enzymes themselves. The FDA hydrolytic activity appeared to be the index most related with the other biological characteristics tested in this study and, for this reason, can be considered the most effective index for putting in evidence relationships existing between the different biological characteristics in the soils investigated.