Microalgal Enzymes with Biotechnological Application

The aim of this thesis was to address the current state of art of microalgal enzymes with biotechnological application, and to expand the current knowledge of enzymes with the potential to find proper application in industry or novel market niches, via their production and biochemical characterization. Available transcriptomic datasets were used to identify possible enzymes of interest. Specifically, after a review of the available scientific literature (Chapter 1) my experimental activities focused on different enzymes identified in the transcriptome of the marine diatom Cylindrotheca closterium. Studied enzymes were two di-n-butyl phthalate hydrolases (CcDBPH1-2), putatively involved in phthalate degradation (Chapter 2), two platelet activating factor- acetylhydrolase (CcPAF-AH1-2), involved in lipid metabolism and inflammation modulation, which role in diatom is still unknown (Chapter 3), a L-asparaginase (CcASNase), that is an enzyme with a well-supported market niche in cancer treatment and acrylamide mitigation (Chapter 4). The experimental pipeline of CcDBPH1-2, starting from transcript identification till protein functional activity studies, was completed and reached the stage of protein purification and activity assay. In addition, different bioinformatics analyses and cc_dbph1-2 expression by RT-qPCR in C. closterium grown with different concentrations of DBP were also performed confirming evidences of the protein bioremediation activity. The expression of CcPAF-AH1-2 was hampered by the strong toxic effect that recombinant proteins production induced in E. coli cells. However, the mutation of the catalytic serine 243 of CcPAF-AH2 allowed enzyme accumulation in 1L cultures. Finally, the expression of CcASNase was successful in BL21 (DE3) and Rosetta 2 (DE3) E. coli strains. However, the low amount of produced protein will need further optimization of the growth conditions. Overall, this PhD thesis represents a contribution to the growing number of studies aimed to unravel the application of microalgae as an eco-friendly and eco-sustainable source of novel enzymes for human applications

Andic soils and catastrophic mudflows in Italy: morphological and hydropedological evidences

In Italy rapid landslides are the most frequently occurring natural disasters and, after earthquakes, cause the highest number of victims. In this contribution we attempt to prove that there exist a tight connection between the presence of a specific soil type, namely andic soils, and the occurrence of the main catastrophic mudflows and debris flows occurred in Italy in the last decades. The study was performed by means of an integrated pedological and hydrological analysis on the detachment crowns of some of the most important catastrophic mudflows and debris flows occurred in Italy in the last decades and involving/evolving surface soils. The results at both regional (Campania) and National (Italy) scale clearly show that despite the large variability of the environmental settings of the studied sites there are indeed some striking homogeneous soil features in the detachment crowns including (i) soil morphology, (ii) andic features ranging from high to moderate, (iii) high water retention throughout a large range of pressure heads. Results seem to reveal clear cause-effect evidences between andic soils and the investigated catastrophic mudflows/debrisflows; this must be related to the unique physical properties of these soils inducing high landslide vulnerability

Intraseasonal Drainage Network Dynamics in a Headwater Catchment of the Italian Alps

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Volcanic soils and landslides: a case study of the island of Ischia (southern Italy) and its relationship with other Campania events

Abstract. An integrated investigation was carried out on the volcanic soils involved in the landslide phenomena that occurred in 2006 at Mt. Vezzi on the island of Ischia (southern Italy). Chemical (soil pH, organic carbon content, exchangeable cations and cation exchange capacity, electrical conductivity, Na adsorption ratio and Al, Fe and Si forms), physical (particle and pore size distribution, pore structure), hydrological (soil water retention, saturated and unsaturated hydraulic conductivity), mineralogical and micromorphological analyses were carried out for three soil profiles selected in two of the main head scarps. The studied soils showed a substantial abrupt discontinuity in all the studied properties at the interface with a buried fine ash layer (namely, the 2C horizon), that was only marginally involved in the sliding surface of the landslide phenomena. When compared to the overlying horizons, 2C showed (i) fine grey ash that is almost pumice free, with the silt content increasing by 20 %; (ii) ks values 1 order of magnitude lower; (iii) a pore distribution concentrated into small (15–30 μm modal class) pores characterised by a very low percolation threshold (approximately 15–25 μm); (iv) the presence of expandable clay minerals; and (v) increasing Na content in the exchange complex. Most of these properties indicated that 2C was a lower permeability horizon compared to the overlying ones. Nevertheless, it was possible to assume this interface to be an impeding layer to vertical water fluxes only by the identification of a thin (6.5 mm) finely stratified ash layer, on top of 2C, and of the hydromorphic features (e.g. Fe / Mn concretions) within and on top of the layer. Although Mt. Vezzi's soil environment has many properties in common with those of other Campania debris-mudflows (e.g. high gradient, north-facing slope, similar forestry, and volcanic origin of the parent material), the results of this study suggest a more complex relationship between soil properties and landslides and emphasise the role of vertical discontinuities as noteworthy predisposing factors

Retroperitoneal pararenal isolated neurofibroma: report of a case and review of literature

The neurofibroma is a tumour of neural origin. This kind of neoplasm, though, is generally skin located. Rare cases in deep organs or in the peritoneal cavity are also reported in the literature. There are two types of neurofibromas, localized and diffuse; the latter is associated with von Recklinghausen disease and always occurs together with skin neurofibromas. Here we report the case of a 47-year-old man affected by retroperitoneal neurofibroma, but not associated with von Recklinghausen disease. A computed tomography (CT) scan described a retroperitoneal pararenal lesion with no clear involvement of adjacent viscera. We describe the diagnostic modality, treatment planning and the timing of treatment of this neoplasm, reviewing also the literature

Intraseasonal Drainage Network Dynamics in a Headwater Catchment of the Italian Alps

In the majority of existing studies, streams are conceived as static objects that occupy predefined regions of the landscape. However, empirical observations suggest that stream networks are systematically and ubiquitously featured by significant expansion/retraction dynamics produced by hydrologic and climatic variability. This contribution presents novel empirical data about the active drainage network dynamics of a 5 km2 headwater catchment in the Italian Alps. The stream network has been extensively monitored with a biweekly temporal resolution during a field campaign conducted from July to November 2018. Our results reveal that, in spite of the wet climate typical of the study area, more than 70% of the observed river network is temporary, with a significant presence of disconnected reaches during wet periods. Available observations have been used to develop a set of simple statistical models that were able to properly reconstruct the dynamics of the active stream length as a function of antecedent precipitation. The models suggest that rainfall timing and intensity represent major controls on the stream network length, while evapotranspiration has a minor effect on the observed intraseasonal changes of drainage density. Our results also indicate the presence of multiple network expansion and retraction cycles that simultaneously operate at different time scales, in response to distinct hydrological processes. Furthermore, we found that observed spatial patterns of network dynamics and unchanneled lengths are related to the underlying heterogeneity of geological attributes. The study offers novel insights on the physical mechanisms driving stream network dynamics in low-order alpine catchments

Evaluating stream CO2 outgassing via drifting and anchored flux chambers in a controlled flume experiment

Carbon dioxide (CO2) emissions from running waters represent a key component of the global carbon cycle. However, quantifying CO2 fluxes across air-water boundaries remains challenging due to practical difficulties in the estimation of reach-scale standardized gas exchange velocities (k(600)) and water equilibrium concentrations. Whereas craft-made floating chambers supplied by internal CO2 sensors represent a promising technique to estimate CO2 fluxes from rivers, the existing literature lacks rigorous comparisons among differently designed chambers and deployment techniques. Moreover, as of now the uncertainty of k(600) estimates from chamber data has not been evaluated. Here, these issues were addressed by analysing the results of a flume experiment carried out in the Summer of 2019 in the Lunzer:::Rinnen - Experimental Facility (Austria). During the experiment, 100 runs were performed using two different chamber designs (namely, a standard chamber and a flexible foil chamber with an external floating system and a flexible sealing) and two different deployment modes (drifting and anchored). The runs were performed using various combinations of discharge and channel slope, leading to variable turbulent kinetic energy dissipation rates (1.5 x 10(-3) epsilon < 1 x 10(-1) m(2) s(-3)). Estimates of gas exchange velocities were in line with the existing literature (4 < k(600) < 32 m(2) s(-3)), with a general increase in k(600) for larger turbulent kinetic energy dissipation rates. The flexible foil chamber gave consistent k600 patterns in response to changes in the slope and/or the flow rate. Moreover, acoustic Doppler velocimeter measurements indicated a limited increase in the turbulence induced by the flexible foil chamber on the flow field (22 % increase in 8, leading to a theoretical 5 % increase in k(600)). The uncertainty in the estimate of gas exchange velocities was then estimated using a generalized likelihood uncertainty estimation (GLUE) procedure. Overall, uncertainty in k(600) was moderate to high, with enhanced uncertainty in high-energy set-ups. For the anchored mode, the standard deviations of k 6 00 were between 1.6 and 8.2 m d(-1), whereas significantly higher values were obtained in drifting mode. Interestingly, for the standard chamber the uncertainty was larger (+ 20 %) as compared to the flexible foil chamber. Our study suggests that a flexible foil design and the anchored deployment might be useful techniques to enhance the robustness and the accuracy of CO2 measurements in low-order streams. Furthermore, the study demonstrates the value of analytical and numerical tools in the identification of accurate estimations for gas exchange velocities. These findings have important implications for improving estimates of greenhouse gas emissions and reaeration rates in running waters

Mars Regolith Simulant Ameliorated by Compost as In Situ Cultivation Substrate Improves Lettuce Growth and Nutritional Aspects

Heavy payloads in future shuttle journeys to Mars present limiting factors, making self-sustenance essential for future colonies. Therefore, in situ resources utilization (ISRU) is the path to successful and feasible space voyages. This research frames the concept of planting leafy vegetables on Mars regolith simulant, ameliorating this substrate’s fertility by the addition of organic residues produced in situ. For this purpose, two butterhead lettuce (Lactuca sativa L. var. capitata) cultivars (green and red Salanova®) were chosen to be cultivated in four dierent mixtures of MMS-1 Mojave Mars simulant:compost (0:100, 30:70, 70:30 and 100:0; v:v) in a phytotron open gas exchange growth chamber. The impact of compost rate on both crop performance and the nutritive value of green- and red-pigmented cultivars was assessed. The 30:70 mixture proved to be optimal in terms of crop performance, photosynthetic activity, intrinsic water use eciency and quality traits of lettuce. In particular, red Salanova® showed the best performance in terms of these quality traits, registering 32% more phenolic content in comparison to 100% simulant. Nonetheless, the 70:30 mixture represents a more realistic scenario when taking into consideration the sustainable use of compost as a limited resource in space farming, while still accepting a slight significant decline in yield and quality in comparison to the 30:70 mixture

TILs in ER+/HER2- breast cancer

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