Urine is considered an ideal source of biomarkers, however in veterinary medicine a complete study on the urine proteome is still lacking. The present work aimed to apply proteomic techniques to the separation of the urine proteome in dogs, cats, horses, cows and some non-conventional species. High resolution electrophoresis (HRE) was also validated for the quantification of albuminuria in dogs and cats. In healthy cats, applying SDS-PAGE and 2DE coupled to mass spectrometry (MS), was produced a reference map of the urine proteome. Moreover, 13 differentially represented urine proteins were linked with CKD, suggesting uromodulin, cauxin, CFAD, Apo-H, RBP and CYSM as candidate biomarkers to be investigated further. In dogs, applying SDS-PAGE coupled to MS, was highlighted a specific pattern in healthy animals showing important differences in patients affected by leishmaniasis. In particular, uromodulin could be a putative biomarker of tubular damage while arginine esterase and low MW proteins needs to be investigated further. In cows, applying SDS-PAGE, were highlighted different patterns between heifers and cows showing some interesting changes during pregnancy. In particular, putative alpha-fetoprotein and b-PAP needs to be further investigated. In horses, applying SDS-PAGE, was produced a reference profile characterized by 13±4 protein bands and the most represented one was the putative uromodulin. Proteinuric horses showed the decrease of the putative uromodulin band and the appearance of 2 to 4 protein bands at higher MW and a greater variability in the range of MW between 49 and 17 kDa. In felids and giraffes was quantified proteinuria reporting the first data for UTP and UPC. Moreover, by means of SDS-PAGE, were highlighted species-specific electrophoretic patterns in big felids and giraffes.

In this study, we investigated the analytical capabilities of DCHQ5, a new fluorescent chemosensor, belonging to the family of diaza-crown-hydroxyquinolines, for the quantitative assessment of total intracellular magnesium content, and its biological applications. We performed a comparative study of DCHQ5 and DCHQ1, the latter being the mother probe of the series, which showed preliminary encouraging results comparable to atomic absorption spectroscopy. We demonstrated that DCHQ5 is able to accurately quantify the total amount of Mg in a very “small” cellular population, by using a simple spectrofluorimetric assay. Furthermore, DCHQ5 demonstrated to be a versatile tool for different applications: its higher intracellular retentions allow to perform cytofluorimetric assays and two-photon confocal microscopy on whole and viable cells; its photochemical characteristic make it excitable in both UV and visible spectra, and the presence of different lifetimes allow to perform fluorescence life time imaging of intracellular Mg. DCHQ5 was also exploited for studying the involvement of magnesium in the commitment of human adipose-derived mesenchymal stem cells (hASCs) with a mixture of hyaluronic, butyric and retinoic acids (HBR). We found that in normal magnesium availability, hASCs precommitment is associated by an increase of total magnesium content during time and by a block in the G2/M phase of the cell cycle. Moreover, our results demonstrated that magnesium deprivation triggers multilineage enrichments of HBR-induced preconditioning of hASCs. The second part of this research was aimed at comparing single cells elemental analysis performed with synchrotron-based fluorescence and cell population analysis carry out by DCHQ5. We exploited innovative techniques of x-ray fluorescence microscopy by using a multimodal approach in order to achieve within the cells the spatial distribution of the concentration of magnesium and fundamental light elements for life. The combination of classical and innovative analytical techniques can shed new light in the comprehension of magnesium homeostasis.

Many organic pollutants are daily released into the environment or pass through the wastewater treatment plants contaminating surface and drinkable water. Part of these pollutants belongs to the category of the emerging organic contaminants since they are still unregulated or in process of regularization. They give cause of concern since they are dangerous for human health and for the survivor of a large number of living organisms. The implementation of wastewater treatment plants against the emerging contaminants is one of the challenges for the enhancement of the water quality, and advanced oxidation processes represent new technologies very promising as tertiary treatments. The research activity carried out during my PhD course focused on degradation test on aqueous solutions contaminated with different class of pollutants such as surfactants, pharmaceuticals and personal care products, both as single compound solutions and mixture of contaminants. The photocatalytic process was studied developing a new pre-industrial pilot plant and testing new TiO2-based photocatalysts in a view of a technological transfer of the photocatalytic methodology. The reuse of a photocatalyst consisting of TiO2 supported on a solid substrates was also investigated with the aim to avoid some problems related to the use of a dispersed catalyst. Photocatalytic process coupled with ultrasounds was also examined in order to speed up the decomposition of the pollutants. The disappearance of every pollutant was followed by HPLC analysis and the mineralization was assessed by the determination of total organic carbon. In the end, the energy consumption related to the processes tested on the mixture of contaminants was calculated in order to establish the best methodology to obtain good degradation rate with reasonable costs.

Weathering steel (WS) describes a class of material, presenting higher corrosion resistance and enhanced mechanical properties than carbon steel when exposed to medium-aggressive environments (rural, urban and light-industrial). The chemical composition of WS, including elements such as Cu, Cr, Ni, Mn, promotes the creation of an adherent passivation layer, which opposes the entry of corrosive species and protects material from further corrosion. However, depending on the exposure environment, the evolution and the composition of patinas can strongly change; the exposure to corrosive atmospheres (i.e., marine sites and industrial/urban areas) destabilizes the patina and reduces corrosion resistance, leading WS to be more susceptible to corrosion and wash-out processes. Consequently, WS releases a certain quantity of alloying metals that could disperse in surrounding areas (soil or water); depending on their chemical form, mobility and bioavailability, some of these elements could represent an hazard to the environment and to living organisms, because of their persistence and potential chronic toxicity. In this work, the issue of atmospheric corrosion of WS was extensively investigated through field and laboratory tests, to evaluate and quantify the effects that surface treatments, exposure geometry, environmental factors and main atmospheric contaminants may have on WS corrosion. Specifically, the characterization of the composition and morphology of corrosion products formed on WS was combined with an innovative approach, that is the quantification of metal dissolution during the early years of exposure, when environmental impact could be greater, and the application of multivariate data analysis. A specific focus was made on the interaction of material with the main aggressive saline ions contained in particulate matter. This work allowed to give a more complete and comprehensive characterization of the environmental behavior of weathering steel and to point out interesting conclusion on the main environmental aspects affecting outdoor corrosion of this material.

Enrollment in K-12 online courses continues to rise substantially each year (Evergreen Education Group, 2015). As the number of students taking courses online increases, the number of parents parenting in online courses also increases. This qualitative exploratory case study, bounded by the online program that was studied, was performed to better understand parents’ perceptions of their roles in online high school mathematics courses. Eighty-seven parents participated in an online questionnaire which elicited both quantitative and qualitative responses. Guided by the major tenets of symbolic interactionism theoretical framework, these responses were combined with data from six interviews to investigate why parents chose to enroll their children in online mathematics courses, their expectations pertaining to the online mathematics course, and their perceived roles and responsibilities in the online mathematics course. Through a detailed process of analyzing the questionnaire and interview data, nine themes emerged: (a) participant parents enrolled their children in online mathematics courses to remove their child from a negative social environment and to avoid distractions in the traditional setting; (b) participant parents want their children to have the flexibility to work ahead of their peers; (c) the school should provide quality curriculum and resources for teachers, students, and parents; (d) teachers should identify and address when students need help; (e) teachers should be available and approachable; (f) students should put forth their best effort; (g) students should ask for help when they experience difficulty understanding a new concept; (h) participant parents monitor to make sure their children are completing assignments and asking for help; and (i) participant parents help their children by re-teaching mathematics concepts or encouraging the child to seek help from others. This study has theoretical and practical significance by adding to literature investigating parental roles in mathematics education and providing insight on the nature of parental involvement in an online high school mathematics program. Consistent with relevant literature (Currie-Rubin & Smith, 2014; Curtis, 2013; Thurber, 2013), results of this study call upon educators to invest in efforts that enhance understanding of parents’ perspectives in an effort to strengthen parental involvement in online mathematics courses. INDEX WORDS: Mathematics, Online learning, Online mathematics, K-12 online learning, Virtual learning, Parental involvement, Parental engagement, Parental roles, Interactions, Teacher responsibilities, Student responsibilities, Success, Perceptions of success, Flexibility

La biochimica clinica dei rettili e dei cheloni in particolare, non ha ottenuto al momento lo stesso livello di attenzione rivolto ai mammiferi. Con il presente lavoro viene proposta la valutazione dei più importanti parametri ematologici e biochimici in campioni di sangue di Testudo hermanni al fine di facilitare l'interpretazione dei dati di laboratorio e la diagnosi di eventuali patologie. A questo scopo, sono stati calcolati gli intervalli di riferimento di questi parametri in individui clinicamente sani e sono state analizzate le influenze di fattori ambientali e fisiologici. Sono state inoltre determinate le concentrazioni di alcuni importanti elementi chimici sia essenziali che non essenziali. Su campioni di chirottero del genere Tadarida teniotis sono state determinate le concentrazioni di PCB DL, PCB NDL, PCDD/F, PFAS e di elementi chimici essenziali e non, al fine di valutare: il possibile livello di contaminazione in relazione alla specie e all’habitat, la prevalenza e/o il rapporto delle classi di composti nel caso di una possibile contaminazione, la eventuale fonte di contaminazione. A quanto ci risulta, il nostro lavoro rappresenta il primo tentativo di analizzare su vasta scala le concentrazioni di questi inquinanti in una popolazione di pipistrelli residenti in un’area urbana. Tra i vari contaminanti esaminanti in questo studio, il Pb e le diossine possono costituire un serio problema per Tadarida teniotis. / Hermann’s tortoise, Testudo hermanni, is currently on the IUCN red list of endangered species. Reptile medicine uses laboratory analyses to evaluate health status, but data on hematology, blood biochemistry and protein electrophoresis are limited in tortoises.The purposes of this study were to establish the most important hematological and biochemical parameters for health assessment in Hermann’s tortoises, calculate the reference intervals and evaluate the impact of environmental and physiological factors.The reference intervals defined in the present study can be considered a useful tool for clinical pathologists and researchers working in tortoise medicine and conservation. In Europe bat populations are gradually declining due to environmental pollution; in this thesis we have studied the concentrations of many contaminants including essential and non essential trace elements, dioxins, PCB DL and NDL and PFAS in tissues of juveniles Tadarida teniotis. The high concentrations of Pb and dioxin determined are indicative of acute evnironmental exposure.

An acceptable agreement permits interchangeability of the instruments. For this purpose, we have investigated the agreement of several clinical instruments frequently used in clinical practice with their laboratory counterpart. We have estimated the agreement between a point-of-care blood gas analyzer (i-Stat, Abaxis) and a bench-top blood gas analyzer (Nova, Biomedical) in venous samples from Hermann’s tortoises. We have estimated the agreement between a point-of-care chemistry analyzer (VetScan VS2, Abaxis) and a laboratory analyzer (Olympus AU400, Olympus Co.) in venous samples from Hermann’s tortoises. We have estimated the agreement between portable blood glucose meters (Accu-Chek, Aviva; AlphaTrak 2, Abbott) and a laboratory analyzer (Dimension EXL, Siemens) in venous samples from client-owned rabbits. We have estimated the agreement between point-of-care bench-top glucose measurement (VetScan VS2, Abaxis) and a laboratory analyzer (Dimension EXL, Siemens) in venous samples from client-owned rabbits. Beyond method comparison and validation, reference interval determination for common laboratory testing is required to allow the clinician to discriminate individuals that are different from the remaining population for a certain parameter. We have calculated reference intervals for blood gas in Hermann’s tortoises. We have calculated reference intervals for protein electrophoresis in Hermann’s tortoises. We have described normal hematology in Hermann’s tortoises. We have calculated reference intervals for clinical chemistry in Hermann’s tortoises. We have calculated reference intervals for aldosterone in ferrets. Based on our results, animal species requires individual validation of laboratory methods and reference intervals. Lack of consideration of these findings may result in clinical misdiagnosis and improper treatment of animals.

Sediments are the final repositories for most pollutants, which can accumulate and remain in the sedimentary matrix for long periods of time and may accumulate through the food web, affecting marine biota, aquatic-dependent wildlife, and ultimately human health. Thus, a wide-ranging work monitoring different groups of contaminants in sediments is a key tool for a comprehensive understanding of contaminants behavior and identification of the overall environmental quality and possible threatens to the whole marine ecosystem. POPs, among which PCBs and DDTs rank first, polycyclic aromatic hyrocarbons (PAHs), and selected groups contaminants of emerging concern (CECs) (fragrances, UV filters, endocrine disruptors) were the objective of this work. First, PCB congeners were analyzed in sediment cores and recent sediments along the north-south axis of the western Adriatic Sea. Chronological records of PCB concentrations and homologue patterns were assessed, supporting the reconstruction of PCBs fluxes and total inventories. Then, the spatial distribution and fate of selected legacy and emerging compounds were investigated in surface sediments sampled along the Adriatic mud-wedge and in deep-sea regions from the southern Adriatic basin, providing the first extensive dataset and also the evaluation of potential ecotoxicological risk of target emerging contaminants in sediments from the Adriatic Sea. Finally, the analyzes of selected legacy contaminants (PCBs, DDTs and PAHs) in sediment cores from coastal and deep-sea sediments from the Adriatic Sea supported the comparison between these two areas and further reinforces the hypothesis on the transfer of contaminants from contaminated areas in the northern Adriatic to the deep southern Adriatic basin. The present thesis is the summary of the work carried out in the last three year of the PhD program “Scienze ambientali: tutela e gestione delle risorse naturali” and contains three manuscripts (two under review and one in preparation), and the main conclusions regarding this PhD research project.

Trabajo de Investigación Requisito para optar al Título de Cirujano Dentista / Autor no autoriza el acceso a texto completo de su tesis en el Portal de Tesis Electrónicas / Introducción: La Periodontitis Apical Asintomática (PAA) corresponde a una patología inflamatoria causada por microorganismos que invaden los canales radiculares del diente. Se caracteriza por la destrucción de los tejidos periapicales en presencia de varios mediadores inflamatorios. La metaloproteinasa de matriz extracelular-12 (MMP-12), expresada en varias patologías inflamatorias crónicas, tiene la capacidad de degradar varios componentes de la matriz extracelular (MEC) tales como elastina, fibronectina y colágeno. También es capaz de activar a otras MMPs con actividad gelatinasa y colagenasa, y dada su función, podría estar involucrada en la patogénesis de la PAA. El objetivo de nuestro trabajo fue determinar la localización tisular de MMP-12 en biopsias de quistes radiculares inflamatorios (QRI), granulomas periapicales (GPA) y en tejido periapical sano You are using demo version Please purchase full version from www.technocomsolutions.com Materiales y Métodos: En este estudio descriptivo se incluyeron 10 sujetos con diagnóstico clínico de PAA e indicación de extracción y 5 sujetos con diagnóstico clínico de Ligamento Periodontal Sano (LS) e indicación de extracción por ortodoncia, con diagnóstico anátomo-patológico de GPA (n=5), QRI (n=5) y LS (n=5). Posteriormente se analizó la inmunolocalización de la MMP-12 mediante inmunohistoquímica para su posterior observación por microscopía óptica. Resultados: De un total de 15 muestras, la MMP-12 se detectó en los 5 QRI y 5 GPA estudiados, mientras que no se detectó en las muestras de LS. En ambos tipos de lesiones periapicales, MMP-12 fue inmunolocalizada principalmente en el infiltrado inflamatorio. En GPA, se observó inmunopositividad en células compatibles con macrófagos, mastocitos, y plasmocitos; mientras que en QRI, MMP-12 se inmunolocalizó en el revestimiento epitelial e infiltrado inflamatorio, particularmente en plasmocitos. Conclusiones: MMP-12 se inmunolocaliza en el infiltrado inflamatorio de GPA y QRI, mientras que no se identificó en LS y por tanto, podría estar involucrada en la patogénesis de la PAA en asociación con el proceso inflamatorio periapical.

Introdução. A paracoccidioidomicose (PCM) é uma micose sistêmica crônica, endêmica na América Latina, principalmente Brasil, sendo a oitava causa de morte entre as doenças infecciosas crônicas recorrentes. A PCM infecção é caracterizada por uma resposta Th1, a forma aguda por um perfil misto da resposta Th2/Th9, enquanto na forma crônica caracteriza-se pelo perfil Th17/Th22. A ocorrência e gravidade da PCM humana podem também estar associadas a fatores genéticos como os polimorfismos dos genes de citocinas. Objetivos. 1. Descrever a frequência dos polimorfismos de (SNPs) IFNG +874 T/A, IL12B 3\' UTR +1188 A/C e IL12RB1 11014 A/G no éxon 7 em pacientes e controles; 2. Investigar a associação entre esses polimorfismos e as diferentes formas clínicas da micose; 3. Verificar se há associação entre esses polimorfismos e a secreção das citocinas IFN-y, IL-12p40 e IL-12p70. Materiais e Métodos. 143 pacientes com PCM foram incluídos (40 com a forma aguda, 100 com a forma crônica multifocal e 17 unifocal). Critérios de inclusão: ter doença ativa (DA) comprovada por exame micológico ou histopatológico positivo ou presença de anticorpos anti-Paracoccidioides brasiliensis (>= 1/32 por contraimunoeletroforese) ou ter doença curada/tratada (CT) quando comprovada anteriormente pelos critérios de DA e atualmente com títulos de anticorpos estáveis e <= 4 em dois períodos com intervalo >= 6 meses. Analisaram-se os SNPs IFNG pela técnica de PCR-ARMS (\"Polymerase Chain Reaction - Amplification Refractory Mutational System\"), IL12B e IL12RB1 por RFLP (\"PCR-Restriction Fragment Lenght Polymorphism\"). Para a dosagem de citocinas foram utilizadas as técnicas de ELISA (n=29) e CBA (\"Cytometric Bead Array\"; n= 18), sendo considerados estatisticamente significantes, os valores de p < 0,05 para os testes de x2 e o teste de Kruskal-Wallis, com pós-teste de Dunn. Resultados. O genótipo AA do SNP IL12RB1 foi mais frequente na forma crônica multifocal e o genótipo AG, na forma unifocal masculina (p= 0,048). À análise desta forma clínica entre ambos os sexos, o genótipo AG foi também mais frequente no sexo masculino (p= 0,009). Segundo a etnia, foi demonstrada diferença estatisticamente significante nas frequências dos genótipos e alelos dos SNPs IFNG e IL12RB1 (p < 0,05). Em relação às formas clínicas da PCM, houve similaridade nas frequências dos genótipos e alelos dos SNPs estudados. Quanto aos níveis das citocinas, para os SNPs IFNG, IL12B e IL12RB1, maiores níveis de secreção de citocinas, frente a PHA, foram registrados nos grupos CT e CO em relação ao DA, sugerindo relação com a evolução da doença e com a imunossupressão já descrita na doença ativa. Conclusão. Não houve associação entre os SNPs IFNG, IL12B e IL12RB1 e as diferentes formas da doença quando todos os pacientes foram analisados; no sexo masculino, sugere-se que o genótipo AA esteja associado à doença crônica mais disseminada (IL12RB1). Houve diferença significante entre as etnias nos SNPs IFNG e IL12RB1, sugerindo-se a ampliação do número de pacientes em determinadas etnias e na forma clínica unifocal para melhor compreensão dessas associações / Introduction. Paracoccidioidomycosis (PCM) is a systemic chronic mycosis, endemic in Latin America, mainly in Brazil where it is the eighth cause of death among chronic recurrent infectious diseases. PCM infection is characterized by the Th1 immune response, the acute form, by a mixed Th2/Th9 profile, while the chronic form is characterized by Th17/Th22 profile. The occurrence and severity of human PCM can also be associated with genetic factors such as polymorphisms on genes of cytokines. Objectives. 1. To describe the frequencies of the single nucleotide polymorphisms (SNPs) IFNG +874 T/A, IL12B 3\'UTR +1188 A/C and IL12RB1 11014 A/G on exon 7, on patients with PCM and non-PCM controls; 2. To investigate the association between those SNPs and the different clinical forms of PCM. 3. To verify the possible association between those SNPs and the secretion of the cytokines IFN-?, IL-12p40 and IL12p70. Materials and Methods. 143 patients with PCM were included (40 with acute form, 100 with multifocal chronic form and 17 unifocal). Inclusion criteria: active disease (DA) proved by fungal identification on direct microscopy/histopathology or culture, or presence of antibodies antiParacoccidioides brasiliensis ( >= 1/32 by counterimmunoelectrophoresis) or cured/treated disease (CT) when previously proved by criteria of DA and present stable antibodies titles =6 months in between. The SNP IFNG was analyzed by PCR-ARMS (Polymerase Chain Reaction - Amplification Refractory Mutational System) and the SNPs IL12B and IL12RB1 by PCR-RFLP (PCR-Restriction Fragment Length Polymorphism). The levels of cytokines were detected by ELISA (n= 29) and CBA (Cytometric Bead Array; n= 18) and values of p < 0.05 for ?2 test and Kruskal-Wallis\' test, with Dunn\'s post-test were considered statistically significant. Results. The AA genotype of SNP IL12RB1 was the most frequent in the multifocal chronic form while the AG was more frequent in men with the unifocal chronic form of PCM (p = 0.048). On this clinical form in the comparison between genres, the AG genotype was also more frequent in men (p= 0.009). On ethnicity, it was demonstrated statistical difference between the frequencies of genotypes and alleles of SNPs IFNG and IL12RB1 (p < 0.05). In the comparison between the clinical forms of PCM, the frequencies of genotypes and alleles of the evaluated SNPs were similar. On the levels of cytokines, for SNPs IFNG, IL12B and IL12RB1, increased levels of cytokines were observed with PHA on the CT and CO groups compared with DA, suggesting a connection with the evolution of the disease and the previously described immunosuppression during active disease. Conclusion. There was no association between the SNPs IFNG, IL12B and IL12RB1 and the different forms of PCM when all patients were analyzed; among men, it is suggested that the AA genotype of IL12RB1 is associated with a more disseminated chronic disease. There was a significant difference between the ethnicities on SNPs IFNG and IL12RB1, being the latter also associated with the chronic form in men. The increase in the number of patients in certain ethnic groups and in the unifocal clinical form of PCM might help the better understanding of these associations