Role of organic acids and phytonutrients as natural alternatives to antibiotics in supporting intestinal functionality

The gastrointestinal tract (GIT) represents the major portion of the body that interfaces with the external environment, with the double function of food processing and line of defense of the body. Numerous components support and regulate the barrier function of the GIT, such as tight junctions (TJs), cytokines, commensal and pathogenic microorganisms, and other systems of the organism, as the endocannabinoid system (ECS). The ECS can control several gastrointestinal functions, as well as the regulation of intestinal inflammation. Failure of the intestinal barrier function triggers an increase of the concentration of pro-inflammatory cytokines and leads to a reduction in intestinal functionality. This thesis aimed to explore the potential of natural compounds as a new alternative approach to antibiotics not only as antimicrobial, but also supporting intestinal maturation and integrity, and as immune-boosting agents. Different experiments were performed to evaluate the potential of nature-identical compounds (NICs), organic acids (OAs), and essential oils (EOs) to support and fight various stressful stimuli. In vitro, a well characterized blend of NICs and OAs were able to improve TJs and transepithelial electrical resistance (TEER) in an intestinal cell line, exerting an anti-inflammatory potential. EOs enhanced TEER and TJs mRNA levels, with a reduction of paracellular permeability, showing antioxidant and antimicrobial properties. In vivo, thymol modulates the gene expression of ECS and gut chemosensing in the GIT of piglets, where the precise localization of the cannabinoid receptors was immunohistochemically confirmed, suggesting an anti-inflammatory potential. In conclusion, natural alternative molecules represent an effective alternative to support or replace the classical pharmacological prophylaxis. These alternative molecules act not only as antimicrobial agents, but also exerted a crucial role in supporting the intestinal barrier function, preventing oxidative stress, and reducing inflammation. Moreover, thymol seems able to modulate the ECS, representing a novel frontier to support animal health and productivity.

Nanocomposites based on poly(caprolactone)-chitosan electrospun nanofibers and metallic oxide nanoparticles. Their potential use to adsorption and photocatalytic degradation of organic contaminants

Pollution of water bodies is one of the most common environmental problems today. Organic pollutants are one of the main drawbacks in this natural resource, among which the following stand out long-lived dyes, pharmaceuticals, and pesticides. This research aims at obtaining nanocomposites based on polycaprolactone-chitosan (PCL-CS) electrospun nanofibers (NFs) containing TiO2 nanoparticles (NPs) for the adsorption and photocatalytic degradation of organic pollutants, using Rhodamine B as a model. The fabricated hybrid materials were characterized by FT-IR, TGA, DSC, SEM, TEM, tensile properties, and the contact angle of water drops. The photoactivity of the NFs was investigated using a batch-type system by following UV-Vis absorbance and fluorescence of rhodamine B (RhB). For this purpose, TiO2NPs were successfully ex-situ incorporated into the polymer matrix promoting good mechanical properties and higher hydrophilicity of the material. The results showed that CS in the NFs increased the absorption and degradation of RhB by the TiO2NPs. CS attracted the pollutant molecules to the active sites vicinity of TiO2NPs, favoring initial adsorption and degradation. In other words, a bait-hook-and-destroy effect was evidenced. It also was demonstrated that the sensitization of TiO2 by organic dyes (e.g., perylene derivative) considerably improves the photocatalytic activity under visible radiation, allowing the use of low amounts of TiO2. (≈0.05 g/1 g of fiber). Hence, the current study is expected to contribute with an environmentally friendly green alternative solution.

Origin and Nature of Radio-Loud Broad Absorption Line Quasars

The origin of Broad Absorption Line Quasars (BAL QSOs) is still an open issue. Accounting for ~20% of the QSO population, these objects present broad absorption lines in their optical spectra generated from outflows with velocities up to 0.2 c. Nowadays, the hypotheses about their nature are principally related to orientation or evolutionary scenarios. In the first one, absorption lines are produced by outflows originated by the accretion disk, basically present in all QSOs, but seen only when they intercept the line of sight. In the second hypothesis, BAL QSOs would be young or recently re-fueled QSOs, still ejecting their dust cocoon. In this case orientation would not play a role, since the absorption features would be produced by spherically ejected matter. In this work we present the results of a multi-frequency study of a Radio-Loud BAL QSO sample, and a comparison sample of Radio-Loud non-BAL QSOs. We performed observations from radio to Near-Infrared, aiming at collecting useful informations about the orientation, the age, and the morphologies of these objects. Various techniques have been applied, including local and continental radio interferometry, single dish observations and spectroscopy. The comparison with the non-BAL QSO sample allows us to conclude that no particular orientation is present in BAL QSOs. Moreover, various morphologies and ages can be found, analogously to "normal" QSOs. Thus, the solution to this astrophysical problem seems not to reside in a peculiarity of the BAL QSO subclass with respect to non-BAL QSOs, since both the studied models do not completely explain the observed characteristics. Further experiments with future instrumentation will allow us to underline useful differences and test the physical conditions in BAL QSOs.

Cellular and molecular effects of cross reacting material 197 (CRM197) on HT-29 colon cancer cell line

Cross Reacting Material 197(CRM197) is a Diphteria toxin non toxic mutant that had shown anti-tumor activity in mice and humans. CRM197 is utilized as a specific inhibitor of heparin-binding epidermal growth factor (HB-EGF), that competes for the epidermal growth factor receptor (EGFR), overexpressed in colorectal cancer and implicated in its progression. We evaluated the effects of CRM197 on HT-29 human colon cancer cell line behaviour and, for CRM197 recognized ability to inhibit HB-EGF, its possible effects on EGFR activation. In particular, while HT-29 does not show any reduction of viability after CRM197 treatment, or changes in cell cycle distribution, in EGFR localization or activation, they show a change in gene expression profile analyzed by microarray. This is the first study where the CRM197 treatment on HT-29 show the alteration of a specific and selected number of genes.

Mitochondrial inheritance and germ line determination in a bivalve species with Doubly Uniparental Inheritance (DUI)

Mitochondria are inherited maternally in most metazoans. However, in some bivalves, two mitochondrial lineages are present: one transmitted through eggs (F), the other through sperm (M). This is called Doubly Uniparental Inheritance (DUI). During male embryo development, spermatozoon mitochondria aggregate and end up in the primordial germ cells, while they are dispersed in female embryos. The molecular mechanisms of segregation patterns are still unknown. In the DUI species Ruditapes philippinarum, I examined sperm mitochondria distribution by MitoTracker, microtubule staining and TEM, and I localized germ line determinants with immunocytochemical analysis. I also analyzed the gonad transcriptome, searching for genes involved in reproduction and sex determination. Moreover, I analyzed an M-type specific open reading frame that could be responsible for maintenance/degradation of M mitochondria during embryo development. These transcripts were also localized in tissues using in situ hybridization. As in Mytilus, two distribution patterns of M mitochondria were detected in R. philippinarum, supporting that they are related to DUI. Moreover, the first division midbody concurs in positioning aggregated M mitochondria on the animal-vegetal axis of the male embryo: in organisms with spiral segmentation this zone is not involved in further cleavages, so aggregation is maintained. Moreover, sperm mitochondria reach the same embryonic area where germ plasm is transferred, suggesting their contribution in male germ line formation. The finding of reproduction and ubiquitination transcripts led to formulate a model in which ubiquitination genes stored in female oocytes during gametogenesis would activate sex-gene expression in the early embryonic developmental stages (preformation). Only gametogenetic cells were labeled by in situ hybridization, proving their specific transcription in developing gametes. Other than having a role in sex determination, some ubiquination factors could also be involved in mitochondrial inheritance, and their differential expression could be responsible for the different fate of sperm mitochondria in the two sexes.

Media activism on and off line by Italian adolescents and young adults

From several researchers it appears that Italian adolescents and young people are grown up with commercial television which is accused to contain too much violence, sex, reality shows, advertising, cartoons which are watched from 1 to 4 hours daily. Adolescents are also great users of mobile phones and spend a lot of time to use it. Their academic results are below the average of Ocse States. However the widespread use of communication technology and social networks display also another side of adolescents who engage in media activism and political movement such as Ammazzateci tutti!, Indymedia, Movimento 5 Stelle, Movimento No Tav. In which way does the world economic crisis -with the specific problems of Italy as the cutting founds for school, academic research and welfare, the corruption of political class, mafia and camorra organisation induce a reaction in our adolescents and young people? Several researches inform us about their use of internet in terms of spending time but, more important, how internet, and the web 2.0, could be an instrument for their reaction? What do they do online? How they do it? Which is the meaning of their presence online? And, has their online activity a continuity offline? The research aims are: 1. Trough a participant observation of Social Network profiles opened by 10 young active citizens, I would seek to understand which kind of social or political activities they engage in online as individuals and which is the meaning of their presence online. 2. To observe and understand if adolescents and young people have a continuity of their socio-political engagement online in offline activities and which kind of experiences it is. 3. Try to comprehend which was (or which were) the significant, learning experiences that convinced them about the potential of the web as tool for their activism.

Clinical outcome and biological characteristics of Chronic Myeloid Leukemia patients treated with nilotinib front-line

The present work reports the outcome of the GIMEMA CML WP study CML0811, an independent trial investigating nilotinib as front-line treatment in chronic phase chronic myeloid leukemia (CML). Moreover, the results of the proteomic analysis of the CD34+ cells collected at CML diagnosis, compared to the counterpart from healthy donors, are reported. Our study confirmed that nilotinib is highly effective in the prevention of the progression to accelerated/blast phase, a condition that today is still associated with high mortality rates. Despite the relatively short follow-up, cardiovascular issues, particularly atherosclerotic adverse events (AE), have emerged, and the frequency of these AEs may counterbalance the anti-leukemic efficacy. The deep molecular response rates in our study compare favorably to those obtained with imatinib, in historic cohorts, and confirm the findings of the Company-sponsored ENESTnd study. Considering the increasing rates of deep MR over time we observed, a significant proportion of patients will be candidate to treatment discontinuation in the next years, with higher probability of remaining disease-free in the long term. The presence of the additional and complex changes we found at the proteomic level in CML CD34+ cells should be taken into account for the investigation on novel targeted therapies, aimed at the eradication of the disease.

Kinematics of local and high-z galaxies through 3D modeling of emission-line datacubes

The kinematics is a fundamental tool to infer the dynamical structure of galaxies and to understand their formation and evolution. Spectroscopic observations of gas emission lines are often used to derive rotation curves and velocity dispersions. It is however difficult to disentangle these two quantities in low spatial-resolution data because of beam smearing. In this thesis, we present 3D-Barolo, a new software to derive the gas kinematics of disk galaxies from emission-line data-cubes. The code builds tilted-ring models in the 3D observational space and compares them with the actual data-cubes. 3D-Barolo works with data at a wide range of spatial resolutions without being affected by instrumental biases. We use 3D-Barolo to derive rotation curves and velocity dispersions of several galaxies in both the local and the high-redshift Universe. We run our code on HI observations of nearby galaxies and we compare our results with 2D traditional approaches. We show that a 3D approach to the derivation of the gas kinematics has to be preferred to a 2D approach whenever a galaxy is resolved with less than about 20 elements across the disk. We moreover analyze a sample of galaxies at z~1, observed in the H-alpha line with the KMOS/VLT spectrograph. Our 3D modeling reveals that the kinematics of these high-z systems is comparable to that of local disk galaxies, with steeply-rising rotation curves followed by a flat part and H-alpha velocity dispersions of 15-40 km/s over the whole disks. This evidence suggests that disk galaxies were already fully settled about 7-8 billion years ago. In summary, 3D-Barolo is a powerful and robust tool to separate physical and instrumental effects and to derive a reliable kinematics. The analysis of large samples of galaxies at different redshifts with 3D-Barolo will provide new insights on how galaxies assemble and evolve throughout cosmic time.

Predictive And Prognostic Role Of Biological Markers In Neuroendocrine Neoplasia And Evaluation Of Activity And Safety Of Second Line Treatments In Neuroendocrine Carcinoma Patients

BACKGROUND Neuroendocrine neoplasia (NEN) are divided in well differentiated G1,G2 and G3 neuroendocrine tumors (NETs) and G3 neuroendocrine carcinomas (NECs). For the latter no standard therapy in second-line is available and prognosis is poor. METHODS Primary aim was to evaluate new prognostic and predictive biomarkers (WP1-3). In WP4 we explored the activity of FOLFIRI and CAPTEM as second-line in NEC patients in a multicenter non-comparative phase II trial RESULTS In WP1-2 we found that 4 of 6 GEP-NEC patients with a negative 68Ga-PET/CT had a loss of expression of RB1. In WP3 on 47 GEP-NENs patients the presence of DLL3 in 76.9% of G3 NEC correlate with RB1-loss (p<0.001), negative 68Ga-PET/CT(p=0.001) and a poor prognosis. In the WP4 we conducted a multicenter non-comparative phase II trial to explore the activity of FOLFIRI or CAPTEM in terms of DCR, PFS and OS given as second-line in NEC patients. From 06/03/2017 to 18/01/2021 53 out of 112 patients were enrolled in 17 of 23 participating centers. Median follow-up was 10.8 (range 1.4 – 38.6) months. The 3-month DCR was 39.3% in the FOLFIRI and 32.0 % in the CAPTEM arm. The 6-months PFS rate was 34.6% ( 95%CI 17.5-52.5) in FOLFIRI and 9.6% (95%CI 1.8-25.7) in CAPTEM group. In the FOLFIRI subgroup the 6-months and 12-months OS rate were 55.4% (95%CI 32.6-73.3) and 30.3% (CI 11.1-52.2) respectively. In CAPTEM arm the 6-months and 12-months OS rate were 57.2% (95%34.9-74.3) and 29.0% (95%10.0-43.3). The miRNA analysis of 20 patients compared with 20 healthy subjects shows an overexpression of miRNAs involved in staminality , neo-angiogenesis and mitochontrial anaerobic glycolysis activation. CONCLUSION WP1-3 support the hypothesis that G3NECs carrying RB1 loss is associated with a DLL3 expression highlighting a potential therapeutic opportunity. Our study unfortunately didn’t met the primary end–point but the results are promising

Study of Bone Biomineralization in 2D and 3D Cultures of a Human Osteosarcoma Cell Line As Osteoblast-like Model

This PhD project focuses on the study of the early stages of bone biomineralization in 2D and 3D cultures of osteoblast-like SaOS-2 osteosarcoma cells, exposed to an osteogenic cocktail. The efficacy of osteogenic treatment was assessed on 2D cell cultures after 7 days. A large calcium minerals production, an overexpression of osteogenic markers and of alkaline phosphatase activity occurred in treated samples. TEM microscopy and cryo-XANES micro-spectroscopy were performed for localizing and characterizing Ca-depositions. These techniques revealed a different localization and chemical composition of Ca-minerals over time and after treatment. Nevertheless, the Mito stress test showed in treated samples a significant increase in maximal respiration levels associated to an upregulation of mitochondrial biogenesis indicative of an ongoing differentiation process. The 3D cell cultures were realized using two different hydrogels: a commercial collagen type I and a mixture of agarose and lactose-modified chitosan (CTL). Both biomaterials showed good biocompatibility with SaOS-2 cells. The gene expression analysis of SaOS-2 cells on collagen scaffolds indicated an osteogenic commitment after treatment. and Alizarin red staining highlighted the presence of Ca-spots in the differentiated samples. In addition, the intracellular magnesium quantification, and the X-ray microscopy on mineral depositions, suggested the incorporation of Mg during the early stages of bone formation process., SaOS-2 cells treated with osteogenic cocktail produced Ca mineral deposits also on CTL/agarose scaffolds, as confirmed by alizarin red staining. Further studies are underway to evaluate the differentiation also at the genetic level. Thanks to the combination of conventional laboratory methods and synchrotron-based techniques, it has been demonstrated that SaOS-2 is a suitable model for the study of biomineralization in vitro. These results have contributed to a deeper knowledge of biomineralization process in osteosarcoma cells and could provide new evidences about a therapeutic strategy acting on the reversibility of tumorigenicity by osteogenic induction.

Development and characterization of cutting-edge equipment for EMC measurements and advanced power line filter design

This Doctoral Thesis aims at studying, developing, and characterizing cutting edge equipment for EMC measurements and proposing innovative and advanced power line filter design techniques. This document summarizes a three-year work, is strictly industry oriented and relies on EMC standards and regulations. It contains the main results, findings, and effort with the purpose of bringing innovative contributions at the scientific community. Conducted emissions interferences are usually suppressed with power line filters. These filters are composed by common mode chokes, X capacitors and Y capacitors in order to mitigate both the differential mode and common mode noise, which compose the overall conducted emissions. However, even at present days, available power line filter design techniques show several disadvantages. First of all, filters are designed to be implemented in ideal 50 Ω systems, condition which is far away from reality. Then, the attenuation introduced by the filter for common or differential mode noise is analyzed independently, without considering the possible mode conversion that can be produced by impedance mismatches, or asymmetries in either the power line filter itself or the equipment under test. Ultimately, the instrumentation used to perform conducted emissions measurement is, in most cases, not adequate. All these factors lead to an inaccurate design, contributing at increasing the size of the filter, making it more expensive and less performant than it should be.