Dissertations / Theses on the topic 'Setters (Dogs)'

In the present research project I explored the effects of old age on cognitive and emotional aspects of the dog-owner relationship. To this aim, three studies were conducted, which focused on a) the detection of age-related cognitive impairment, b) the assessment of selective social attention skills in adult and aged dogs and c) the effect of age and of impaired cognitive functions on the dog-owner emotional bond. Although the existence of age-related cognitive impairment has been acknowledged in veterinary medicine, methodological issues limited the amount of scientific research in this aspect of companion dogs’ ageing. To address this problem, in the first study (pp. 10-47), we presented the development of a new method for the detection of cognitive decline in aged dogs. We employed a multiple approach, combining a questionnaire-based evaluation of dogs’ behaviour (Cognitive Evaluation Scale, CES) with the assessment of dogs’ memory and reversal learning abilities in a spatial cognition task. Application of these methods to a population of adult and aged dogs resulted in the detection of significant differences between the two age groups, with aged dogs obtaining lower CES scores and showing a worse performance in the spatial cognition task. A correlation between the two parameters was an important further indication of the validity of the method. Compared to other currently available tools, the present approach could have characteristics of higher sensitivity and specificity, providing practical advantages, for both future research in the field and for the clinical assessment of cognitive decline. A further aspect that received scarce scientific interest in aged dogs, is their attention towards humans, a phenomenon that could play a critical role in the expression of socio-cognitive abilities and in the adaptability of dogs into inter-specific groups. In the absence of suitable methods, we developed a behavioural task protocol to assess the quantitative and qualitative characteristics of dogs’ selective attention in an inter-specific context. In the first part of this second study (pp. 48-75), we presented the validation of this procedure, performed on a sample of adult pet dogs. The results showed a selective response in dogs simultaneously exposed to the owner and to an unfamiliar person, demonstrating that the pattern of attention is dependent on the strength of the dog-human relationship. Moreover, the task allows discriminating ‘passive’ (the simple orientation of senses to a stimulus) from ‘active’ (involving cognitive processing) forms of attention. In the second part of the study, the application of this method to a sample of aged dogs revealed a detrimental effect of age on the level of selective attention towards the owner. Old dogs had a higher propensity for ‘passive’ attention, being less capable than adults to retain information about social stimuli when these disappear from their visual field. The third study (pp. 75-98) dealt with the effect of old age on the nature of the dog-owner emotional bond. Sharing similarities with the human infant-mother relationship, the dog-owner bond has been described as a form of attachment: an enduring relationship with a particular other, which becomes apparent through distinctive behaviours, expressed under stress situations. Despite human research emphasizes the importance that emotional bonds might hold in the well being of aged individuals, the study of this aspect was neglected in the dog. We investigated the effect of old age on dogs’ attachment behaviours, using the Strange Situation Test, a method originally designed to elicit attachment responses in human infants. Our results show that old age does not affect the expression of ‘secure’ attachment (confidence at the presence of the object of attachment and distress at separation). However, we saw in old dogs an alteration in the expression of other behaviours (play and passive behaviours), which could represent a different emotional dimension of the dog-owner attachment (insecure-ambivalent). Since part of the dogs’ sample that was included in the present study had also participated in the previous studies, when possible the effect of cognitive impairment and selective attention on the dog-owner emotional bond was analyzed.
Il presente progetto di ricerca indaga gli effetti dell’età senile su aspetti pertinenti alla sfera cognitiva ed emozionale della relazione cane-proprietario. A tal fine sono stati condotti tre studi indipendenti, incentrati su: a) la rilevazione di deficit cognitivi legati all’età, b) la valutazione delle capacità di attenzione sociale selettiva in cani adulti e anziani e c) gli effetti dell’età anziana e del decadimento cognitivo sulle manifestazioni di attaccamento al proprietario. Benché l’esistenza di un declino cognitivo età-dipendente sia un’evenienza riconosciuta in medicina veterinaria, problemi di carattere metodologico hanno limitato la produzione scientifica su questo aspetto dell’invecchiamento nel cane da compagnia. Nel tentativo di ovviare a queste limitazioni, nel primo studio (pp. 10-47), è presentato lo sviluppo di un nuovo metodo per la rilevazione di deficit cognitivi legati nel cane anziano. Il metodo si basa su un approccio multiplo, che combina l’uso di scale di valutazione cognitiva, basate su questionari (Cognitive Evaluation Scale, CES) con la valutazione diretta delle capacità memoniche e di ‘apprendimento inverso’ dei cani in un test di cognizione spaziale. L’applicazione di questi metodi a una popolazione di cani da compagnia adulti e anziani è esitata nella rilevazione di differenze significative tra i due gruppi di età. I cani anziani ottengono punteggi peggiori nella scala di valutazione cognitiva e mostrano compromesse capacità di apprendimento nel test di cognizione spaziale. La correlazione tra i due parametri rappresenta un’importante indicazione della validità del metodo. Paragonato ad altri strumenti attualmente disponibili, questo approccio potrebbe garantire maggiore sensibilità e specificità, apportando un importante vantaggio pratico, sia in ambito di ricerca, che quale strumento di valutazione diagnostica in ambito clinico veterinario. Un ulteriore aspetto che ha ricevuto scarso interesse scientifico nel cane è l’attenzione rivolta all’uomo, un fenomeno che potrebbe rivestire un ruolo critico nell’espressione di capacità socio-cognitive e nell’integrazione del cane in gruppi sociali misti. In assenza di metodi appropriati, è stato sviluppato un test comportamentale per la valutazione delle caratteristiche quantitative e qualitative dell’attenzione selettiva del cane in un contesto interspecifico. Nella prima parte di questo secondo lavoro (pp. 48-75), presento lo sviluppo e la validazione di questa procedura, effettuata su un campione di cani adulti. I risultati mostrano una risposta selettiva nell’attenzione che i cani rivolgono al proprietario o a una persona estranea, cui sono simultaneamente esposti. Inoltre, il protocollo permette di discriminare forme di attenzione ‘attiva’ (con il coinvolgimento di processi cognitivi) e ‘passiva’ (cioè il semplice orientamento visivo). L’applicazione di questo metodo a un campione di cani anziani, nella seconda parte dello studio, ha rivelato un effetto negativo dell’età anziana sul livello di attenzione selettiva rivolta al proprietario. I cani anziani, inoltre, dimostrano una maggiore propensione per forme di attenzione ‘passiva’ essendo meno in grado, rispetto agli adulti, di ritenere informazioni riguardo allo stimolo sociale all’uscita di questo dal campo visivo. Nel terzo studio (pp. 75-98) è stato affrontato l’effetto dell’età anziana sulla qualità del legame emotivo cane-proprietario. In studi precedenti il legame cane-proprietario è stato assimilato all’attaccamento infante-madre. L’attaccamento definisce un legame duraturo con uno specifico individuo, che si manifesta attraverso comportamenti espressi in condizioni di stress. Benché ricerca in ambito umano abbia dimostrato la rilevanza della sfera affettivo-emotiva nella salvaguardia del benessere delle persone anziane, questo aspetto risulta scarsamente investigato nella specie canina. Nel presente studio il legame cane-proprietario è stato studiato mediante lo Strange Situation Test, un protocollo originalmente sviluppato per evidenziare comportamenti di attaccamento nel bambino in età infantile. I risultati mostrano come l’età anziana non alteri le manifestazioni comportamentali di attaccamento ‘sicuro’ (sicurezza in presenza e stress alla separazione dal proprietario). Tuttavia si evidenzia nel cane anziano un’alterazione nell’espressione di altri comportamenti (gioco e comportamento passivo), che potrebbero riflettere una diversa dimensione emotiva dell’attaccamento (insicuro-ambivalente). Poiché parte dei cani inclusi in questo studio avevano preso parte anche ai precedenti studi, quando i dati lo consentivano è stato indagato l’effetto del declino cognitivo e di differenze nelle capacità attentive sull’espressione dell’attaccamento cane-proprietario.

In the last decades several studies have been investigated the behavior and cognition of dogs (Canis familiars) especially from socio-cognitive aspects. We already gained a solid knowledge on how dogs communicate with humans and conspecific partners, how they are able recognize, interpret and rely on social-communicational cues in their everyday lives, thus continuously perceiving a flow of visual information. However, the fundaments of how they process these visual information are still missing from the literature. The current PhD project have been developed around three independent studies, all of them investigating visual cognition in dogs (and for one case, also in humans) in the domain of perceiving motion.
Negli ultimi decenni sono stati studiati diversi studi sul comportamento e sulla cognizione dei cani (familiari di Canis), specialmente da aspetti socio-cognitivi. Abbiamo già acquisito una solida conoscenza su come i cani comunicano con gli umani e con i partner conspecifici, come sono in grado di riconoscere, interpretare e fare affidamento su segnali di comunicazione sociale nella loro vita quotidiana, percependo continuamente un flusso di informazioni visive. Tuttavia, i fondamenti di come elaborano queste informazioni visive sono ancora mancanti dalla letteratura. L'attuale progetto di dottorato è stato sviluppato intorno a tre studi indipendenti, tutti studiati sulla cognizione visiva nei cani (e per un caso, anche negli esseri umani) nel campo del percepire il movimento.

Diarrhea is a common problem in recently weaned and young dogs. If the separation from the litter occurs at a time when the immune system is not fully competent or whether as the puppy had a lower colostrum consummation or a bad alimentation, this combination of events increases the susceptibility of the puppy or young dog to a variety of infections and gastrointestinal disturbances. Bacterial pathogens, such as enteropathogenic Escherichia coli and enterotoxigenic E. coli are also associated with enteric disease in young dogs. Our study about bovine colostrum supplementation aimed to know its effects on recently weaned puppies at 32 days of age. The puppies were divided into 2 groups each composed of 3 and 4 animals, the control one (CT) and the treatment one (TG) randomly. The experimental groups belonged to the same litter. The involved puppies were fed with the same basal diet. A treatment was represented by administering bovine colostrum once a week through a specific oral syringe. Feed intake was registered daily. Individual body weight and average daily gain were measured weekly. We found that its principal benefit was an increased gut flora such as Lactobacillus spp. versus E.coli. in treated puppies. The purpose of the second study was to evaluate the role of the integration of an alternative source of fiber, i.e. microcrystalline cellulose, in relation to its potential for action in the intestine, in the diet of dogs suffering from acute gastrointestinal symptoms. For the test we took into consideration twenty subjects, ten of which received a simple industrial diet, while the remaining ten were given the same industrial insoluble fiber (microcrystalline cellulose) enriched diet in such quantity that the final ration contained crude fiber of 10% dry matter. All subjects featured a symptomatic gastrointestinal clinical manifestation characterizing variable output of feces, soft feces, formed feces, the feces without definite form until liquid stools. The third study was about chronic diarrhea as a consequence of food intolerance. The dietary management of these cases varies according to the underlying disease. Generally, the animal should receive a diet with moderate levels of fiber and fat divided in into small portions during the day. The objective of this diet was to facilitate intestinal regulation, beneficially modify the composition and metabolic activity of the intestinal microbiota and exclude antigens from the diet. In our cases the patents showed a good response to these elimination homemade diets.

Leishmaniasis is a parasitic disease caused by Leishmania spp. Once transmitted to the host, the parasite, being an obligate intracellular pathogen, localizes primarily inside macrophages, where it starts replication. Interferon γ (IFN-γ) is one of the most important mediators produced by Th1 subpopulation and it is responsible for macrophages activation that could lead to parasite control. Neopterin belongs to the chemical group known as pteridines. It is synthesised also by macrophages upon stimulation with interferon-γ and is indicative of immune system activation. Measurement of N concentrations in body fluids like blood, cerebrospinal fluid or urine provides information about the cell-mediated (T helper cells type 1 Th1) immune sistem activation in humans; assuming that, the present study aimed at investigating if N could also be considered as an indicator of a protective reaction (Th1-mediated) of the host against Leishmania, and then if it could be used clinically as prognostic factor in leishmaniotic patients or as helpful parameter in monitoring treated patients, as investigated in men. For these purposes, the study was divided into two phases independent of each other. The first phase of the study aimed to at investigating the possible correlations between sera N values and antibody titers (dosed by immunofluorescence antibody test IFAT) in 74 dogs, both seronegative and seropositive. In the second one, 5 patients affected by leishmaniasis and in treatment for it, were followed for a period of 24 months comparing N values to other parameters (e.g. IFAT, PCRs, citology on fine needle aspiration (FNA) of lymphnodes). Even in the absence of a statistically significant correlation between N values and IFAT titers, serum N appeared to be an interesting marker possibly able to provide an indirect value of Th1/Th2 ratio in leishmaniotic patients. It should also be further investigated its possible role in the early detection of ill patients possibly predisposed to develop renal failure and/or to relapse.

Strain elastography is useful in differentiating benign from malignant lesions in human medicine. The aim of this study was to assess the repeatability and reproducibility of elastography in the evaluation of canine splenic lesions, mammary lesions and superficial nodules and to verify if they can be differentiated in benign or malignant. Twenty-four dogs presenting a single splenic hypoechoic lesion, underwent B-mode ultrasonographic and elastographic evaluation. The Strain Ratio (SR) and Hardness Value (HV) were calculated for all the lesions. The SR and HV of benign lesions were statistically different from those of malignant lesions (p<0,05). Malignant lesions tended to have a SR >1,5 (p=0,0003; Se 75%, Sp 100%, PLR 0, NLR 0.25, PPV 100%, NPV 88.24%) and an HV > 70% (p=0,001; Se 100%, Sp 75%, PLR 4, NLR 0, PPV 66,67%, NPV 100%). The correlation between SR >1,5 and malignancy was statistically significant (OR, 80,6; p=0,0067) as the one between HV >70% and malignancy (OR, 47,22; p=0,013). This technique was repeatable and reproducible (CV 0,08 ± 0,05 and K=1 for the SR; CV 0,09 ± 0,06 and K=0,67 for the HV). Strain elastography can differentiate malignant from benign canine hypoechoic splenic lesions less then 4 cm width. Twenty-one dogs presenting mammary, underwent B-mode ultrasonographic and elastographic evaluation. Each mammary lesion was then classified according to the Tsukuba score, the presence /absence of infiltration on the elastogram and to the elastogram/Bmode ratio. The Pearson’s chi-square test was used to assess the presence of a statistically significant difference between the benign or malignant tumors according to the Tsukuba score results. The Fisher’s exact test was used to assess the presence of statistically significant difference between the benign or malignant tumors according to the elastogram/B-mode ratio. Values of p ≤0,05 were considered significant. The K of Cohen was used to verify the agreement between the elastosonographic assessment of infiltration and the histological confirmation of tumors invasion. The Pearson’s chi-square test did not reported significant differences between groups (p=0,278) even considering the assessment of the elastosonographic infiltration. The K of Cohen presented a value of 0,384 corresponding to a poor agreement. The assessment of the elastogram/B-mode ratio was not possible. Finally the Strain Elastography was as an applicable technique in evaluating mammary neoplasms in our sample, demonstrating their elastographic patterns; but it was clinically not useful in differentiating benign from malignant lesions. Sixty nodular skin lesions from 36 canine patients were evaluated using B-mode ultrasonographic and elastographic evaluation. Each lesion was classified using 2 score systems by a software and then subjectively by three operators. The K of Cohen was used to verify the agreement between the elastosonographic assessment of the three operators. The Student t-test was used to assess whether there was a statistically significant difference between benign/malignant considering the hardness value calculated using the software. The Pearson chi-square test was used to assess the existence of statistically significant differences between benign and malignant lesions considering the two score proposed above. Values of p < 0,05 were considered significant. The result obtained in the Student's t test and Pearson’s chi square test allow us to say that there is a statistically significant difference between benign and malignant lesions evaluated by HV and score system respectively (p<0,05). Considering the inter-observer agreement we can see that among all three operators there is a very good correlation in score evaluation. From the average values calculated for the weighed K, we can see that there is a better agreement among operators than between observer and the objective score. The results obtained in this study are encouraging about the possibility of applying the elastosonography the nodular lesions of the skin. We have seen that this technique is able to differentiate benign and malignant lesions due to the change of elasticity of the tissues involved by malignant pathological processes. the very good agreement between operators and the good agreement between operators and objective score demonstrate the feasibility of this technique.

The long stay of dogs in the shelters is often a welfare problem for the animals, a cost to society and an ethical problem. It is therefore important to prevent the abandonment and increase the success rate of adoptions. The aim of this study was to identify, through the application of a protocol for behavioural assessment, an adoption indicator easily definable for dogs coming into the shelters, that allows to estimate the ease with which the animal will be adopted. 593 sheltered dogs (age 2 months to 18 years) were involved in this study over a period of three years starting from March 2010. The data were divided into three groups, A, B and C. The physical (breed, age, sex, size and physical pathologies) and behavioural data of each dog were assessed by a trained examiner using an appropriate test. In the Group A the results of the characteristics presenting a significant difference distribution between adopted or not adopted dogs, were utilized to calculate a synthetic number that we named ''IDA'' (r/s). A standardized post adoption telephonic follow-up (3 months after the adoption) was performed for investigate whether the adoptions were satisfactory for owners. Subsequently, in order to verify if the hypothesis we created was reliable, the adoption indicator (IDAr and IDAs) has been applied to C and B. The data were analysed by chi square tables (SPSS 12,0, 2003). The dogs scoring more then the mean value (rounded) of the adopted dogs of the Group A were defined as easy adoptable (H, high probability to be adopted), the dogs scoring less then the mean value of the not adopted dogs were set out as hardly adoptable (L), and the other as of medium adoptability (M). In the Group A the proportion between adopted and not adopted dogs (within 3 months after recovery) were significant for IDAr between H and M or L (P minus 0,01) and between M and L (P minus 0,05); The IDAs significant differences were between H and L (P minus 0,01) and M and L (P minus 0,05). A first way to validate the reliability of the adoption indicator was to explore the rate of adoption in the animals of Group A during the period of one year after recovery There are a strong link between IDA score and adoption rate in the period (P minus 0,01), more clear for IDAs, with more significant differences between High and Low scored dogs (P minus 0,01) than between Hig and Medium (P minus 0,05 for IDAr; P minus 0,01 for IDAs). The satisfaction rate of owners was 98 %. In the Group C the percentage of adopted dogs with H IDAr or IDAs were significantly different from those of M and L IDAr/IDAs dogs (P minus 0,01) but not between M and L. Therefore as for the Group A the results show that an H IDA corresponds to a high percentage of adoption. Therefore in the Group C the value of the adoption indicator (IDAr/s), calculated according to the same procedure designed for the Group A, did not different significantly from that of Group A (P minus 0,05). The application of the IDA scores to the dogs of group C was analyzed also during the year after recovery and as for the Group A the results confirm that the correlation between IDA and adoption rate is maintained in the long term. In the Group C the satisfaction rate of owner was 98,75%. The results of the correlation analysis between the periods elapsed by dogs in the kennel show that the IDA scores correlate negatively with the period of time from recovery in the kennel to adoption or, for the not adopted dogs, till one year after recovery. In order to verify the reliability of the data was evaluated the mean of the time elapsed in the kennel for dogs with H, M or L IDAr and H, M and L IDAs in the three groups. Dogs with a H IDA tend to stay in shelter less time than those with a lower IDA. The IDA as calculated by us seems to meet the indications for a good test : Feasibility, Reliability and Validity.

Summary Social referencing and understanding of human emotional expressions in dogs. Introduction The literature on dog cognition and in particular social cognition has grown incredibly in recent years, however no studies have focused on the process of Social referencing in dog-human communication and interaction, and only few preliminary studies have investigated dogs’ understanding of human emotions. Social referencing is a process characterized by the use of another person’s perceptions and interpretation of a situation to form one’s own understanding and guide action. Human infants use this process to go beyond the information given by an informant to construct a more general interpretation of the meaning of a stimulus and they can successfully link the emotions expressed to a referent object. Social referencing has been investigated not only in humans, but also in a few primate species, namely chimpanzees and barbary macaques, although with mixed results. In fact, results from primate studies are overall ambiguous, with the strongest evidence of social referencing coming from human-raised chimpanzees, where it is exhibited with their human caretaker. There is, however, more consistent evidence that non-human primates can refer conspecific and human emotional expressions to a referent object. The dog-human relationship is a very special one and the more recent literature shows that the dog-human bond is similar in many respect to an infantile attachment. Furthermore, studies suggest that dogs are sensitive to a number of behaviours potentially revealing the person’s attentional states; can follow a number of human referential cues, and, preliminary evidence, seems to suggest that they can also discriminate between some expressions of human emotions. Finally there is also some evidence that dogs can communicate intentionally and referentially with humans (although this is still a mater of some controversy). Considering the above, dogs are particularly good candidates for the comparative investigation of social referencing and of the ability to understand human’s emotional expressions as referring to specific objects. Four studies are presented in this thesis which aimed to investigate Social referencing in dog-human dyads and dogs’ capacity to understand the referential nature of a person’s emotional message. The aims of the present research were to add to the literature on dogs’ socio-cognitive abilities and of human-dog communication by studying (1) the presence of Social referencing, both referential looking and behavioural regulation, in dogs towards humans, (2) the potential selectivity of this process, based on the relationship with the informant (owner vs. stranger), (3) the effect of a particular kind of training experiences (i.e. water rescue training) on this process, and (4) the ability of dogs to refer different emotional expressions toward two objects. To answer these questions we carried out four different experiments: Study 1 Social referencing in dog-human dyads was investigated using the “new object paradigm”. As no study has been carried out on dogs on this topic so far, we set up a new procedure suitable to this species, that was similar to the one used in the infant. In particular in our “new object paradigm” we presented dogs with a new and potentially scary object (a fan with plastic ribbons attached to it) in presence of their owner as the informant. The aim was to evaluate whether, in a social referencing paradigm, dogs would show referential looking and behavioural regulation toward the owner acting as the informant and hence approach the object more having witnessed a positive vs. a negative message. We tested 75 dog-owner dyads at the Canis Sapiens Lab of the University of Milan. There were two different groups of dogs: in one group the owner was delivering a positive emotional message towards the object, whereas in the second group the owner delivered a negative emotional message. The results of this first study showed that dogs, like human infants, use referential looking towards a familiar person (their owner) in a situation of ambiguity. However, differently from infants, dogs showed no clear evidence of behavioural regulation after receiving an emotional message from the owner. Study 2 Since the results obtained in the first study showed a clear presence of referential looking in dogs, but not clear evidence of behavioural regulation toward the owner, we modified the testing procedure to further evaluate behavioural regulation. We investigated the presence of Social referencing in dogs with the same potentially scary object (a fan with plastic ribbons attached on it) in presence of their owner vs. a stranger. The study had two main aims. First, since results obtained in the first study provided only unclear evidence of behavioural regulation toward the object, we wanted to see whether using a procedure more closely resembling the one used with infants we would be able to find evidence of behavioural regulation. The second aim was to assess the influence of the informant’s identity on social referencing. Selectivity is an important aspect of children’s Social referencing: they seem to use referential looking toward both a familiar and an unfamiliar person, but they regulate their behaviour only when a familiar person is the informant or when she/he is present in the experimental set-up together with the unfamiliar informant. We tested 90 dog-owner dyads and there were four groups of dogs: two groups were tested with the owner as the informant (either expressing a positive or a negative emotion), and two were tested with a stranger as informant (either expressing a positive or a negative emotion). Results provided clear evidence that dogs use referential looking not only towards their owner but also towards a stranger, with no difference between the two persons. Furthermore, dogs regulated their behaviour towards the object after receiving a positive or a negative emotional expression by the owner, but not when the stranger was acting as the informant. Study 3 Since in Study 2, besides referential looking, we found a clear evidence of behavioural regulation with the owner, whereas no clear results emerged with the stranger acting as the informant, we tested dogs exposed to a particular type of training: water rescue dogs. In fact, during their training these dogs become used to focusing their attention on unfamiliar person. Thus we investigated the presence of Social referencing in dog-human dyads using the same procedure and scary object as in study 2 but with a stranger acting as the informant. For this study we tested 22 dog-owner dyads: the group of trained water rescue dogs were tested with their owners in Naples (Università degli Studi di Napoli “Federico II”), while the breed, age and sex matched control group was tested in Milano at the Canis Sapiens Lab of the University. Dogs of both groups showed referential looking toward the stranger and this result supports those obtained in study 2. However, highly trained water rescue dogs did not change their behaviour towards the object (behavioural regulation) when the stranger was exhibiting a positive emotional expression towards it. Surprisingly, the control (untrained) group dogs did change their behaviour toward the ambiguous object, approaching it more than the trained dogs when the positive emotional message was given by the stranger. These findings are discussed in relation to our previous results (in Study 2) where no such effect was found with the stranger acting as the informant (we believe the likely cause is the breed of dogs tested in Study 3) and in relation to the potential inhibitory effect of training on dogs’ behavioural regulation in this context. Study 4 In the previous three studies dogs were tested always in the presence of only one object (the potentially scary fan), and, although the informant’s message always referred to that object, the goal of these studies was not strictly to assess whether dogs were capable of appropriately referring the human’s emotional expression to the object itself. So far only one study has tested dogs’ ability to attribute a human’s emotional reaction (facial expression and short vocalization) to a specific object (i.e. to grasp that emotions can be referred to something in the outside world). Thus the aim of this study was to assess dog’s understanding of human emotional expression as referential. In particular, we evaluated if dogs can discriminate between three different emotional expressions (fear, happiness and neutral) and whether they have a perception that a human’s (owner vs. stranger) emotional expression can refer to specific objects in the environment. We tested 95 dog-owner dyads at the Canis Sapiens Lab of the University of Milan. We adapted a procedure used with infants, and more recently also with chimpanzees and dogs, in which the informant expresses two different emotions towards two identical (hidden) objects. In a control group, the same procedure is adopted but in the absence of the objects (“no-object” condition). After observing the emotional expressions being conveyed by the informant (owner vs. stranger) dogs were free to approach the objects. Dogs showed a clear preference for the hidden object eliciting the positive emotion, compared to one eliciting the negative one when the owner was acting; on the contrary no preference emerged when the stranger was acting as informant. Furthermore, dogs didn’t show a choice behaviour when the owner was expressing the emotion in the “no-object condition”. A follow-up study, contrasting the positive and negative emotion in turn with a neutral one, showed that dogs tended to approach the object eliciting a positive emotion rather than avoid the one eliciting a negative reaction. Taken together these results, show that dogs do in fact appreciate that an emotional message can relate to a specific object if the owner is the informant but they do not do so if the stranger is the informant. Furthermore, their performance seems to be based on approaching the positive stimulus rather than avoiding the negative one. Together these results suggest that prior experience with their owner using positive emotional expressions, has allowed dogs to associate these emotions to particular objects. Finally, considering the non-specific behaviours exhibited by dogs in the no-object control group, dogs may even have come to expect that an emotional message refers to a specific object. Conclusions The experiments presented in the current thesis reveal some new and interesting aspects of dog social cognition and communication with humans. On the one hand they provide the first evidence of social referencing in dogs: they show that dogs, like infants, can show referential looking and behavioural regulation in ambiguous situations when the emotional message is delivered by the owner, or by a stranger (in presence of the owner). Finally, these findings suggest that dogs have some understanding that emotional expressions are referential, in the sense of being directed to specific stimuli in the environment as has been reported for infants and non human primates.

The aim of echocardiography is to perform the study of the cardiac function, through the evaluation of alterations of anatomy, contractility and flow. The ultrasound techniques used until now to reveal possible alterations of myocardial contractility, such as b-mode, m-mode and Tissue Doppler Imaging (TDI) haven't allowed to obtain complete evaluations. In 2002, Buckberg, analyzing the cardiac structure, demonstrated a complex movement of the heart, determined by the particular helical disposition of the myocardial bundles. The motion is composed by three different components: radial, longitudinal and of torsion. The TDI is dependent from the angle Doppler of incidence, therefore it allows to estimates only movements parallel to the ultrasound beam. Recently, the application of Speckle Tracking , being independent from the angle Doppler, has permitted to improve the study of the cardiac motion. X-StrainTM is a new ultrasound tool by ESAOTE, based on Feature Tracking, that is able to perform the evaluation of the ventricular performance. The aims of this study are to verify the possible application of X-StrainTM in 18 dogs affected by mitral regurgitation, to verify the relationship between ventricular indexes (ejection fraction, shortening fraction, and left atrium diameter- aorta diameter ratio) obtained through traditional techniques (B-MODE, M-MODE), X-StrainTM and the severeness of heart failure in dogs with mitral regurgitation (ISACHC classification), and to verify if X-StrainTM indexes (radial strain and strain rate, endocardial and epicardial circumferential strain and strain rate, radial and rotational displacement of endocardium and epicardium) could have a prognostic value and allow an easier therapeutic control of the patient during the follow-up. The dogs have been evaluated twice (T0 and T1), and the data have been subjected to a statistical analysis. It has been demonstrated a significant positive correlation between the radial strain and strain rate and the ejection an shortening fractions. Besides, to the increase of the radial strain and strain rate, corresponds an improvement of the clinical conditions. The radial strain seems to be less sensible to the volume overload caused by the mitral regurgitation. The endocardial and the epicardial circumferential strain and strain rate have the same trend at T0 and T1. This condition suggests that the motion of torsion could be not influenced by the volume overload. Besides, it has been possible to consider the follow-up of four dogs treated with inotropic medicines, and it has been possible to note their improvement of ventricular contraction. The results therefore, suggest the possibility to use X-StrainTM as a valid ultrasound method in dog to complete the cardiac study, and in particular, to reveal alteration of regional and global myocardial contractility, and to evaluate the complex motion of torsion of heart. It would be interesting to perform a study in a bigger sample of dogs, to obtain physiological data and to verify the eventual different effect of the treatment. X-StrainTM could allow to check the follow-up of dogs, to choose the right therapy.

PDA is one of the most common congenital cardiac malformation in the dog. Echocardiography has been the cornerstone in diagnosing and providing hemodynamic information in a wide variety of disease.Recently, speckle tracking echocardiography has been regarded as a useful tool to assess cardiac function. Aim of the study was to compare cardiovascular hemodynamics in dogs with PDA with healthy control dogs and evaluate any change in cardiac contractility before and after PDA closure. A statistically significant difference was found between standard and advanced echocardiographic techniques, with absolute higher values in dogs with PDA as compared to healthy controls. Cardiovascular hemodynamics were markedly different before and after PDA closure. Based on the results, STE should be considered in echocardiographic assessment of dogs with PDA, as they represent an animal model of congenital left ventricular overload not affected.

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The world of sugar plantation in Brazil, since the conquest and colonization, has been built as a favored subject of European speeches, as per example the 1500-s chronicles of European travelers and settlers, besides the innumerous mentions contained in the Jesuitic Letters, especially those from José de Anchieta and Manuel da Nóbrega, obtaining, in 1711, an enormous visibility with the work Culture and opulence of Brazil, from Father Antonil. Among the most varied speeches of colonial phase, the subject of the Portuguese sugar enterprise in Brazil would surpass the texts from periods after our autonomy, guaranteeing its frequency as much in literary speech as in essayistic and sociological speeches. In the beginning of 20th century, this subject would be retaken as central by the Pernambucan Mário Sette, with the work Sugar plantation lady. The publication of this novelistic speech, in 1921, would make Mário Sette the precursor of Modern School in Northeast and Northeastern Regionalism of 20th century and, consequently, a forerunner of the essayistic speeches of Gilberto Freyre and the novelistic narratives of José Américo de Almeida and José Lins do Rego, literary icons of Northeastern Modernism. Far from losing strength as speech motivation, the sugar society would be retaken as novelistic speech at the end of 20th century with the publication of the work Bonfire of memory, from Francisco Dantas, published in 1991. Knowing the importance of the sugar universe in our several cultural speeches, we undertook a comparative reading between the works Culture and opulence of Brazil, from Father Antonil, and Sugar plantation lady, from Mário Sette, seeking to observe the representative traits that informam and enformam these speeches about the world of sugar plantation in Brazil. To achieve our purpose we take refuge with an interdisciplinary theoretical-methodological approach, more precisely in the basis of Integral Critic outlined by the Brazilian critic Antonio Candido since the beginning of the second period of last century.
O mundo do engenho no Brasil tem, desde o período da conquista e da colonização, se constituído como tema privilegiado dos discursos europeus, a exemplo das crônicas quinhentistas, de viajantes e colonizadores, além das inúmeras menções contidas nas Cartas jesuíticas, especialmente as de José de Anchieta e as de Manuel da Nóbrega, alcançando, em 1711, uma enorme visibilidade com a obra Cultura e opulência do Brasil, do Pe. Antonil. Presente nos mais variados discursos da fase colonial, a tematização da empresa lusitana do açúcar, no Brasil, perpassaria os textos dos períodos que sucedem a nossa autonomia, garantindo sua freqüentação tanto no discurso literário, quanto no discurso ensaístico e sociológico. Nos inícios do século XX, essa temática seria retomada, como centralidade, pelo pernambucano Mário Sette, com a obra Senhora de engenho. A publicação desse discurso romanesco, em 1921, tornaria Mário Sette o precursor do Modernismo no Nordeste e do Regionalismo nordestino do século XX e, consequentemente, num antecipador dos discursos ensaísticos de Gilberto Freyre e das narrativas romanescas de José Américo de Almeida e de José Lins do Rego, ícones literários do Modernismo nordestino. Longe de arrefecer, como motivação discursiva, a sociedade do açúcar seria novamente retomada, como discursividade romanesca, em fins do século XX, com a publicação da obra, Coivara da memória, de Francisco Dantas, publicada em 1991. Cientes da importância do universo açucareiro em nossos mais diversos discursos culturais, empreenderemos uma leitura comparativa entre as obras Cultura e opulência do Brasil, do Pe. Antonil, e Senhora de engenho, de Mário Sette, buscando observar os traços representativos que informam e enformam esses discursos sobre o mundo do engenho no Brasil. Para a realização de nosso propósito, nos valeremos de uma abordagem teórico-metodológica interdisciplinar, mais precisamente dos pressupostos da Crítica Integral, esboçada pelo crítico brasileiro Antonio Candido, desde os inícios da segunda metade do século passado.

The unit research of Camerino executes a study on 9 dogs affected by periodontal disease (PD), of phase 2, 3 and 4, to evaluate the regenerative capacity of the nanohydroxyapatite in the periodontal tissue. The project lasts not more than 24 months. During this period the recruitment of the subjects and the record on the dental sheet relative to the periodontium conditions, supported by a radiographic and iconographic documentation, are effected. The half animals, divided in 3 homogeneous groups of 3 subjects in base on the level of the estimated PD, is subjected to a regenerative therapy of the periodontium (PD phase 2 and 3) and eventually to extraction and method by plant (PD phase 4). The dogs are subjected quarterly to clinical exams and other evaluations both instrumental (RX, TAC, RM, Echocardiography) and of laboratory (hemochrome and eventually hemoculture). The results are then analyzed with a statistical method.

Renal secondary hyperparathyroidism (RHPT) is a common complication in humans and dogs affected with chronic kidney disease (CKD). RHPT is promoted mainly by the loss of nephrons and consequent hyperphosphatemia and its severity increases in parallel to the disease. The identification and management of RHPT are suggested in dogs affected with CKD, to establish promptly appropriate treatments and follow the progression of the disease. However, in clinical practice, the determination of parathyroid hormone (PTH) concentration is not routinely due to limited available and validated method and need of radioimmunoassay. In CKD, data about the presence of RHPT in large populations of dogs and the thorough investigation of the association of PTH and other clinicopathological parameters involved in kidney disease are limited. Moreover, in human medicine, PTH has also been identified as an independent prognostic factor in patients affected with CKD and it is also reported that dysregulation of mineral metabolism leading to RHPT is present also in course of acute kidney injury (AKI), as occurs in CKD. Information about these two aspects in dogs is lacking. Hence, this thesis has been divided into two main parts: the first about analytical aspects and the second about clinical data. The first part was focused on the analytical validation of a new immunoenzymatic method validated in humans for the measurement of canine intact PTH and the establishment of the reference interval in healthy dogs. The second main part of this thesis was aimed to investigate the prevalence of RHPT in dogs affected by CKD and AKI, to compare these populations, and to assess the association between PTH and hematological, biochemical, and urinary parameters. Finally, a survival study has been done to elucidate the possible role of PTH as a survival factor in dogs with CKD. The first part of the study has allowed us to successfully validate the method in dogs, which has showed satisfying accuracy and repeatability, and to demonstrate a storage stability of PTH until 6 months at freezing temperature. A reference interval of 1.42-14.59 pg/mL has been established. The second part has allowed us to confirm the relevant presence of RHPT in CKD, also in early stages, and discover several associations between PTH and some parameters such as creatinine and phosphorus, as known, and others not yet identified, such as potassium, hematocrit, and blood pH. RHPT has been identified also in a high percentage of dogs affected with AKI, though the PTH concentrations were not significantly different between AKI and CKD. The survival analysis has revealed that higher concentrations of PTH corresponded to significantly short survival times compared to the lower concentrations, showing a possible prognostic role of PTH in dogs.

Despite some experiments have been carried out to investigate dog’s visual cognitive abilities and to study the mechanisms underlying how they promptly modify their behaviour while interacting with humans, it is still not clear to which features of the stimuli they mainly attend when solving a task. In addition to physical proprieties, there are different factors that modulate the saliency of a visual stimulus in dogs, such as the previous experience and training received. To investigate whether during the associative learning some aspects of visual information have different salience during the associative learning and the role played by previous experiences, the project was developed around three independent studies. The purpose of the first study was to assess the visual processing of global and local levels of hierarchical stimuli in domestic dogs. Fourteen dogs were trained to recognise a compound stimulus in a simultaneous conditioned discrimination procedure and were then tested for their local/global preference in a discrimination test. As a group, dogs showed a non-significant trend for global precedence, although large inter-individual variability was observed. Choices in the test were not affected by either dogs’ sex or type of stimulus used for training. However, the less time a dog took to complete the training, the higher the probability that it chose the global level of test stimulus. Moreover, dogs which showed a clear preference for the global level in the test were significantly less likely to show positional responses during discrimination training. These differences in the speed of acquisition and response patterns may reflect individual differences in the cognitive requirements during discrimination training. The individual variability in global/local precedence suggests that experience with using visual information may be more important than predisposition in determining global/local processing in dogs. The second study, composed by two experiments, investigated the relevance of visual stimuli using more complex images: human face pictures. Dogs exhibit characteristic looking patterns when looking at human faces but little is known about what the underlying cognitive mechanisms are and how much these are influenced by individual experience. In Experiment 1, seven dogs were trained in a simultaneous discrimination procedure to assess whether they could discriminate a) the owner’s face parts (eyes, nose or mouth) presented in isolation and b) whole faces where the same parts were covered. Dogs discriminated all the three parts of the owner’s face presented in isolation, but needed fewer sessions to reach the learning criterion for the eyes than for both nose and mouth. Moreover, covering the eyes region significantly disrupted face discriminability compared to the whole face condition while such difference was not found when the nose or mouth was hidden. In Experiment 2, dogs were presented with manipulated images of the owner’s face (inverted, blurred, scrambled, grey-scale) to test the relative contribution of part-based and configural processing in the discrimination of human faces. Furthermore, by comparing the dogs enrolled in the previous experiment and seven ‘naïve’ dogs we examined if the relative contribution of part-based and configural processing was affected by dogs' experience with the face stimuli. Naïve dogs discriminated the owner only when configural information was provided, whereas expert dogs could discriminate the owner also when part-based processing was necessary. The present study provides the first evidence that dogs can discriminate isolated internal features of a human face and corroborate previous reports of salience of the eyes region for human face processing. Although the reliance on part-perception may be increased by specific experience, our findings suggest that human face discrimination by dogs relies mainly on configural rather than on part-based elaboration. For a more applied comprehension of the phenomenon and to understand what happens in a more natural situation, the third study explored the characteristic of dog’s attention to natural visual stimuli (humans) during a training session in which different type of exercises and levels of difficulty were requested. Moreover, the quality and quantity of training dogs had received during their lives was taken into account to investigate a possible influence on their attention to humans. As in the second study, the specific effect of training emerged also in the third. Having received a specific training modified dogs’ attention towards the caregiver. The variation of attention pattern did not emerge in a baseline condition but only when the caregiver asked the dog to perform some obedience exercises evidencing that the training had a very context-related effect. In conclusion, dogs seem to acquire more quickly and spontaneously global/configural information of a visual stimulus and when an effect of previous experience such as training is present, this effect is specific and strictly context-related.
Nonostante alcuni esperimenti abbiano indagato quali siano le abilità cognitive visive del cane ed esplorato i meccanismi alla base della sua prontezza nell’adeguare il comportamento durante l’interazione con l’uomo, non è ancora chiaro a quali caratteristiche degli stimoli visivi i cani prestino maggiore attenzione mentre risolvono un compito. Oltre alle proprietà fisiche, sono molti i fattori che possono modulare la salienza di uno stimolo visivo, come per esempio l’esperienza pregressa e l’addestramento ricevuto. Il presente progetto è stato sviluppato su tre studi indipendenti, al fine di studiare quali aspetti dell’informazione visiva abbiano maggior rilevanza durante l’apprendimento associativo e quale ruolo giochi l’addestramento. Il primo studio ha esplorato le caratteristiche della processazione (globale e locale) di stimoli geometrici gerarchici nel cane domestico. Quattordici cani sono stati addestrati a distinguere uno stimolo composto, applicando una procedura di discriminazione simultanea condizionata e la loro preferenza per il livello locale/globale è stata verificata con un test di discriminazione visiva. Il gruppo di cani ha dimostrato una tendenza non significativa per la precedenza globale, sebbene sia stata riscontrata un’ampia variabilità individuale. Le scelte eseguite durante il test non sono state influenzate nè dal genere dei cani, nè dal tipo di stimolo usato durante l’apprendimento. Tuttavia, minore era il tempo impiegato per completare la fase di apprendimento, maggiore era la probabilità che il cane scegliesse il livello gloable dello stimolo visivo durante il test. Inoltre, i cani che hanno dimostrato una preferenza per il livello globale dello stimolo visivo durante il test hanno manifestato, con minor probabilità, risposte posizionali durante l’apprendimento. Queste differenze, emerse sia nella velocità di apprendimento della procedura che negli schemi di risposta durante il test, potrebbero rispecchiare l’esistenza di prerequisiti cognitivi diversi tra i soggetti. La variabilità individuale, riscontrata nella precedenza globale/locale di processazione, suggerisce che l’esperienza nell’uso delle informazioni visive potrebbe essere più importante della predisposizione nel determinare quale sia la tipologia di processazione (globale/locale) usata dai cani. Il secondo studio, composto da due esperimenti, ha indagato quale sia la rilevanza degli stimoli visivi utilizzando immagini più complesse, ossia fotografie di volti umani. I cani esibiscono schemi di attenzione caratteristici quando guardano i volti umani, però si conosce poco dei meccanismi cognitivi sottostanti e quanto questi siano influenzati dall’esperienza individuale. Nel primo esperimento, sette cani sono stati addestrati applicando una procedura di discriminazione simultanea per verificare che fossero in grado di discriminare: a) le parti del volto del proprietario (occhi, naso o bocca) presentate singolarmente; b) le facce intere dove le stesse parti erano state coperte. I cani sono riusciti a discriminare tutte e tre le parti della faccia del proprietario presentate singolarmente, ma, per gli occhi, hanno avuto bisogno di un minor numero di sessioni per raggiungere il criterio di apprendimento stabilito rispetto a naso e bocca. Inoltre, la discriminabilità dei visi è stata inficiata di più quando la regione degli occhi non era visibile, che non quando erano nascosti naso o bocca. Nel secondo esperimento, per testare il contributo relativo della processaziona basata sulle parti o sulla configurazione nella discriminazione di facce umane, sono state usate delle immagini del viso del proprietario manipolate in diversi modi (invertite, sfuocate, ‘scrambled’ - ossia suddivise in parti e riarrangiate in modo casuale - e in scala di grigi). Inoltre, confrontando i sette cani coinvolti nel primo esperimento con sette cani ‘naïve’ è stato esaminato se il contributo relativo della processazione basata sulle parti o sulla configurazione sia stato influenzato dall’esperienza che hanno avuto i cani con gli stimoli raffiguranti le facce. I cani naïve sono riusciti a discriminare il proprietario solo quando l’informazione configurazionale era conservata, mentre i cani esperti sono riusciti a discriminare il proprietario anche quando era necessario processare le immagini basandosi sulle parti. Questo studio prova, per la prima volta, che i cani riescono a discriminare gli elementi interni di un viso umano presentati singolarmente e conferma la salienza della regione degli occhi per la processazione di facce umane. Sebbene un’esperienza specifica possa incrementare la capacità di basarsi sulla percezione delle parti, i risultati suggeriscono che, nel cane, la discriminazione di volti umani si basa principalmente sull’elaborazione della configurazione che delle parti. Per una comprensione più applicata del fenomeno e per capire cosa succede in un contesto più naturale, il terzo studio ha esplorato le caratteristiche dell’attenzione del cane a stimuli naturali (uomo) durante una sessione di addestramento, in cui era richiesta l’esecuzione di esercizi di diverso tipo e difficoltà. Inoltre, sono state prese in considerazione la quantità e la qualità di addestramento ricevuto nel corso della vita dei soggetti per indagare la loro eventuale influenza sulle caratteristiche dell’attenzione prestata all’uomo. Come emerso anche nel secondo studio, è stato riscontrato un effetto specifico dell’addestramento. Aver ricevuto un addestramento specifico influenza l’attenzione prestata dai cani al proprietario. L’attenzione non è variata, infatti, in una condizione di base, ma solo quando il conduttore ha chiesto al cane di eseguire alcuni esercizi di obbedienza, mettendo in luce che l’addestramento ha avuto un effetto contesto-specifico. Per concludere, dai risultati ottenuti nel corso dei tre esperimenti del progetto, sembra che i cani acquisiscano più velocemente e spontaneamente le informazioni visive globali e configurazionali. Quando è presente un effetto dell’addestramento, questo è specifico e strettamente legato al contesto.

After prolonged learning we all have experienced a reduction of alertness, resulting in errors that we would normally not make. Despite this being a common situation in everyday life, the reasons for this phenomenon are unclear. A possible explanation is that the regions of the brain which are involved in the learning, go off-line trying to partially recover. This event is defined as local sleep and it has been detected in animals and sleep-deprived humans performing learning tasks. Local sleep is a sleep-like electrophysiological activity occurring locally, while the rest of the brain is fully awake, and producing performance deterioration. However, since all the studies included both lack of sleep and learning, it is uncertain whether such phenomenon is related to sleep deprivation or if it is the consequence of prolonged learning. Further, local sleep has not been related to electrophysiological changes occurring during the task. This thesis aimed to assess, for the first time in well rested subjects, whether local sleep and performance decline occur because of prolonged learning. Specifically, the goal was to discriminate between sustained practice and learning, as to determine whether learning is required to cause local sleep. Also, a 90-minute nap was evaluated to establish whether sleep is necessary to counterbalance neuronal fatigue and performance decrease. The starting hypothesis was that local sleep is a plasticity-related phenomenon affecting performance and requiring learning to be triggered. Consequently, sleep would be a prerequisite to counterbalance performance and electrophysiological changes. High-Density EEG and behavioral data of 78 healthy young subjects were collected during and after two learning tasks performed for three hours: a visual sequence learning task, and a visuo-motor rotation task, randomly selected. Afterward, subjects were divided in two groups: those who slept for one hour and a half and those who remained awake and quietly rested for the same amount of time before being tested for electrophysiological and behavioral changes. Moreover, to discriminate between the effects of prolonged learning and practice, 11 additional subjects performed a control condition consisting in planar upper limb reaching movements instead of the above-mentioned learning tasks. In detail, the power spectrum of the EEG activity during the task and at rest with eyes opened was divided into five ranges to determine frequency changes of the EEG activity: delta 1 to 4 Hz; theta 4 to 8 Hz; alpha 8 to 13 Hz, beta 13 to 25 Hz, gamma 25 to 55 Hz. Additionally, movement-related beta activity of 35 young subjects was analyzed to find a relationship between task related oscillations and performance indices, as the modulatory activity during practice may reflect plasticity-related phenomena that can describe the occurrence of local sleep. Finally, 13 young subjects were compared to a dataset of 13 older participants who performed planar upper limb reaching movements to determine whether beta oscillations were affected by age. Specifically, beta activity was assessed during reaching movements in different brain regions, in terms of topography, magnitude, and peak frequency. Results demonstrated that sustained learning produced electrophysiological changes both at rest and during the task. In fact, resting state was characterized by a progressive slowing of the EEG activity over areas overlapping with those engaged during the task. Precisely, we detected task-related activity mainly in the high-frequency ranges (gamma and beta right temporo-parietal activity for the visual sequence learning task; alpha and beta activity over a fontal and left parietal areas for the visuo-motor rotation); the same areas were characterized by a progressive increase of the low frequency EEG activity at rest ranging from alpha, beta after one hour of practice, to theta after three one-hour blocks. The control task did not trigger such EEG slowing, as reaching movements without learning did only left an alpha, beta trace in the resting state over a cluster reflecting the motor area contralateral to the movement. Further, continuous learning triggered performance deteriorations only in tests sharing the same neural substrate of the previously performed task. In other words, the visuo-motor learning task only affected performance in a motor test consisting in random reaching movements; conversely, visual sequence learning altered performance on a visual working memory test, but did not influence reaching movements. Also, the control condition did not affect performance in any of the two exercises. Performance decline, learning ability and local sleep were partially renormalized by a 90-minute nap but not by an equivalent period of wake. As such, the global EEG activity, computed as the mean power of all the electrodes, was not affected by either 90 minutes of sleep or quiet wake. However, the regions characterized by low frequency at rest benefited from the sleep period, as the low frequencies content partially decreased after the nap but not after quiet wake. Task related beta activity during motor practice presented similar magnitude and timing patterns in different brain areas, with a progressive increase with practice, in both young and older subjects, despite the older subjects performing slower, less accurate movements. Intriguingly, the motor areas showed a post movement beta synchronization having a peak between 15 and 18 Hz, as opposed to a frontal area that has it between 23 and 29 Hz. Finally, results did not reveal any direct relationship between EEG beta oscillations and performance indices. Altogether, these results indicate that local sleep and performance decrease can be triggered by prolonged learning in well rested subjects; furthermore, some amount of sleep can partially renormalize learning ability, EEG activity and performance. Also, differences in the brainnoscillations during motor activity can express separate processes underlying motor planning, execution and skills acquisition. The present study adds some important knowledge in the field of local sleep; in fact, it suggests that such phenomenon is triggered by sustained learning rather than sleep deprivation, thus being a plasticity-related phenomenon. Finally, the role of sleep on counterbalancing local sleep has been proved, despite additional studies are required to establish whether a full night of sleep rather than a specific amount of time is needed to fully restore learning ability and electrophysiological activity. In conclusion, the present findings are of importance in all the fields where sustained learning is required, such as rehabilitative programs, sport and military trainings, and must be taken into account when plasticity plays a fundamental role in the acquisition of new skills.

Omics techniques have been widely applied to veterinary science, although mostly on farm animal productions and infectious diseases. In canine oncology, on the contrary, the use of omics methodologies is still far behind. This thesis aims to fill in the gap in the molecular background of canine cancer tumors and to apply this new knowledge for the identification of suitable biomarkers mainly based on the identification and quantification of microRNAs. The thesis applied next generation sequencing (NGS) to investigate onco-miRNome and skin microbiota of one of the most relevant, highly metastatic canine malignant tumor, the mast cell tumor. The results of miRNomic have been validated by RT-qPCR to develop molecular tools to enhance the robustness of clinical decision-making. To elucidate the host-tumor interaction, the skin and dermis microbiota of MCT-affected dogs has been investigated as well. This thesis aims also to characterized the proteomic profiles of exosomes purified from plasma of MCT-affected dogs. Due to the pandemic situation, the protocol for exosome purification has been developed, while the proteomic analysis is still ongoing. Finally, a very preliminary evaluation of miRNA dysregulation in canine oral and cutaneous melanoma was carried out. To characterize the epigenetic modulation of MCT microenvironment and to identify a panel of miRNAs dysregulated during the neoplastic process and suitable as biomarkers, the miRNome of canine MCT was profiled on FFPE samples using an NGS approach; the result was then investigated also in saliva samples of MCT-affected dogs. The NGS findings revealed that 63 miRNAs were differentially expressed between MCT and healthy adjacent margins (45 down-regulated and 18 up-regulated). A panel of nine miRNAs was validated by qPCR, of which five were dysregulated in the tumor. Specifically, miR-21 and miR-379 were up-regulated while miR-885, miR-338 and miR-92a were down-regulated in the MCT compare to the healthy margins. A further analysis including the variable of the lymph node involvement in the tumor, revealing that a panel of three miRNAs, miR-21, miR-379, and miR-885, was able to discriminate metastatic from non-metastatic MCTs. The diagnostic potential of the panel was also explored in the saliva of MCT-affected dogs, showing that miR-21 and miR-885 discriminated tumor-affected from healthy dogs; miR-885 could also discriminate the metastatic from the non-metastatic MCTs with good diagnostic potential, make this miRNA attractive for a future diagnostic purpose starting form a less-invasive matrix. The microbiota of the MCT associated with the skin surface and dermis was characterized for the first time investigating alpha diversity which describes the differences of taxa by qualitative and quantitative approaches and beta diversity which describes the differences between groups generating distance matrices between samples. The data showed that a reduction of taxa was present in the tumor skin surface compared with the healthy contralateral part with an increase of Firmicutes phylum and Corynebacteriaceae family. A similar reduction of taxa found out in the tumor dermis compared with the overlying tumor skin surface. These differences exerted an impact also in beta diversity, which highlighted the differences between the samples. The characterization of the core microbiota of the tumor and healthy skin surface and the tumor dermis showed that the healthy skin was characterized by a higher number of ASVs (27 ASVs) compared to the tumor skin surface (12 ASVs) and dermis (16 ASVs) and that 10 ASVs were present only in the tumor site. The data proved that the presence of the tumor interfered in some way with the microbial population, paving the way for further investigations on host-microbiota interaction. Moreover, the reduction of microbiota diversity and taxa was seen to be associated with an unhealthy status. A protocol for the purification of plasmatic exosome from dog affected by MCT was developed to obtain samples with a good degree of purity and suitable for proteomic analysis, that may elucidate the cell-to-cell communication strategy used by MCT to prepare the metastatic niches, and may promote the identification of suitable biomarkers for the MCTs diagnosis and prognosis. The Size Exclusion Chromatography purification approach was identified as the best approach to obtain samples depleted from the most abundant plasmatic protein, including albumin, and exosomes in the right size range and with concertation feasible for further proteomic analysis. Finally, the potential of a target miRNAs panel was also evaluated in canine cutaneous and oral malignant melanoma. The data showed that, among six miRNAs selected from the bibliography, miR-145, miR-365, miR-146a, and miR-425 were differentially expressed in canine melanoma. In detail, miR-145 was down-regulated in cutaneous and oral melanoma and miR-365 was down-regulated in cutaneous melanoma. On the other hand, miR- 146a and miR-425 were up-regulated in cutaneous melanoma. The gene ontology analysis suggested that these miRNAs may have a role in tumor progression and cell proliferation.

Prion protein (PrPC) is a cell surface glycoprotein, anchored by the GPI to the cell membrane. It seems to be involved in some neurodegenerative diseases, but its physiological function is still undefined. Many cellular functions were proposed as PrPC roles, such as the modulation of several signal transduction pathways known to promote cellular survival and the protection against oxidative stress. Nevertheless, the main hypothesis on PrPC role is in copper homeostasis. Copper is an essential micronutrient (EMN) and all EMNs (vitamins and metals) are present in bodies and foods in micromolar concentrations. This feature can easily lead to deficiency. It is also clear that deficiency of EMNs produces overlapping diseases symptoms. Also the uptake pathway (endocytosis) is common for many EMN. Thanks to these reasons we hypothesize that prion protein could allow EMNs uptake. The aim of this work is to study these binding processes and their characteristics. So N-terminal domain of mouse PrPC (mPrP23-109) was recombinant expressed and purified. To study specificity and characteristics of metals binding process, mPrP23-109 was titrated with five first transition serie divalent metals (Mn, Co, Ni, Cu, Zn), at several pH values and it was followed in fluorescence spectroscopy. Furthermore, to define if metal binding could drive structural rearrangements in the prion protein N- terminal domain, we performed structural analysis using circular dicroism (CD). To investigate vitamins stacking between triptophan indolic rings, mPrP23-109 was titrated with four vitamins belonging to B group and titrations were monitored by fluorescence anisotropy, that allows to know the fluorophores average molecular rotational speed. Finally, we also investigated the possibility that prion protein N- terminal domain could interact with membrane mimetic systems. Our findings confirm that prion protein has a functional role in copper homeostasis. We also propose that prion protein, together with copper, plays a key role in integrated endocytic uptake pathway, involving all EMNs.
La protein prionica (PrPC) è una glicoproteina legata alla superficie extracellulare della membrana tramite l’ancora GPI. Ad essa è stata attribuita la causa di alcune malattie neuro degenerative, ma la sua funzione fisiologica non è ancora stata definita. Molte funzioni sono state proposte, tra cui la modulazione di alcune vie di transduzione del segnale che promuovono la sopravvivenza cellulare e la protezione da stress ossidativo, ma l’ipotesi principale è una funzione nell’omeostasi del rame. Il rame è un micro nutriente essenziale (MNE) e tutti gli MNE sono presenti nel corpo e negli alimenti in concentrazioni micromolari; ciò può determinare facilmente una deficinza. È ormai chiaro che i sintomi da deficienza sono comuni a molti MNE. Inoltre anche le vie di assorbimento di molti MNE sono analoghe fra loro, usando vie endocitiche. Grazie a queste evidenze abbiamo ipotizzato che la PrPC possa essere coinvolta nell’assorbimento degli MNE. Lo scopo di questo lavoro è di studiare questi legami e le loro caretteristiche. Per far ciò, è stato espresso e purificato il dominio N-terminale della proteina prionica di topo (mPrP23-109). Per studiare la specificità e le caratteristiche del legame con i metalli, la proteina ricombinante è stata titolata con cinque metalli divalenti della prima serie di transizione (Mn, Co, Ni, Cu, Zn), a diversi valori di pH utilizzando la spettroscopia di fluorescenza. Per definire se le interazioni osservate fossero funzionali, è stata effettuata l’analisi strutturale in dicroismo circolare. Per verificare la proabile interazione tra le vitamine ed i triptofani della proteina, mPrP23-109 è stata titolata con quattro vitamine, appartenenti al gruppo B, ed è stata monitorata la velocità dei fluorofori mediante l’anisotropia di fluorescenza. Infine è stata anche studiata l’interazione tra dominio N-terminale della PrPC con sistemi mietici di membrane. I nostri risultati confermano che la PrPC ha un ruolo funzionale nell’omeostasi del rame. Inoltre proponiamo che la proteina prionica, insieme con il rame, ricopre un ruolo chiave nell’assorbimento integrato degli MNE, via endocitosi.

Photophysical properties of semiconductor quantum dots (QDs), such as broad absorption band and size dependent spectral emission, together with the high effective surface area available for interaction with target chemicals are attractive for applications like chem-/bio-sensors and lab-onchips. In the present work the state of the art of the research field will be discussed, with particular emphasis on the physical and chemical properties of near-infrared (NIR) emitting quantum dots. So far most of the reports of chem-/bio-sensing with QDs are based on colloidal nanocrystals synthesized by wet chemistry methods. Recently, a new approach has been introduced, which made use of Surface QDs (SQDs) grown by epitaxy on solid substrates. These QDs are not suitable to be used in solution, and thus cannot be exploited for application like in vivo imaging and diagnostics. However, the peculiarity that they are directly grown on a semiconductor surface represents a major advantage for chem-/bio-sensor and lab-on-chip design. At least in principle all is needed for a fluorescence based sensor is a light source, the quantum dots and a light detector, and all these components can be easily made with well-developed semiconductor fabrication technology. The presented research has given a contribute to this interesting new perspective. In particular this study is focused on InP SQDs. These island-like nanostructures were synthesized by gas source molecular beam epitaxy (GS-MBE) on In0.48Ga0.52P buffer layer lattice matched to Si doped GaAs substrate. They present a room-temperature NIR photoluminescence (PL). The emission wavelength depends on their dimensions (in the range 750-865 nm). For the present study samples presenting high dot coverage have been synthesized and characterized in term of structural, optical and chemical sensing properties. The most important achievement of this study has been the observation of a reversible PL enhancement when the SQDs were exposed to the vapours of different polar protic solvents. The emission energy and shape were not affected by the solvent vapours, while the photoluminescence intensity depends on solvent vapour concentration (with linear law over a limited concentration range). Besides, a clear correlation between the structural parameters of the SQDs and the response to chemicals has been identified. The presented results showed that InP SQDs are suitable materials for application like optical chemical sensors, bio-sensors and lab-onchip devices.

Recent years have seen a rapid improvement in the techniques for the localization of people buried in avalanches through the use of radio transmitters (transceivers). However, when the avalanche victims are not equipped with transmitter equipment, the use of rescue dogs remains the most effective method for locating them. Although the level of training of rescue dogs is generally good, there remains a considerable margin of improvement. Such an improvement could be achieved through innovative training methods based on the analysis of the dogs performance in a multi-disciplinary context. Among the problems that make this kind of study difficult, there is our lack of knowledge of the chemical and physical variables that govern the diffusion of volatile and semi-volatile organic compounds rising from the buried body to the snow surface. Moreover, we still lack a study of a dog’s olfactory ability to discriminate among the different classes of molecules emitted by the human body (sweat, breath, clothes, etc.) and how these molecules reach the surface after moving through the snow mass. One of the ultimate goals of this study is to select specific olfactory targets that can be used for training avalanche dogs.

During the last decades, the combination of cellular and molecular biology and mechanical engineering, has stimulated the development of new multidisciplinary fields in biomedical sciences, such as tissue engineering and regenerative medicine. The primary goal of these approaches is to provide an alternative to organ and tissue transplantation, by creating viable substitutes for damaged or diseased organs and tissues. Tissue healing is a complex process that involves a cascade of cellular and molecular events, that are mostly shared by the different tissues of the body. Interestingly, the tissue repair process initiates immediately after a traumatic injury and is mediated and controlled by a wide range of cytokines, proteins and growth factors released from platelets upon activation. Consequently, many growth factors have being considered as therapeutic molecules for the repair or regeneration of a wide range of tissues. Although their role has been only partially elucidated, the potential benefit of most growth factors has been demonstrated. In the last few years, the development of platelet-rich preparations has revolutionized the field of regenerative medicine, due to the repair capacities of the platelet-released growth factors, that stimulate and accelerate both soft and hard tissue healing and regeneration. Today, platelet concentrates are used in a wide range of disciplines such as dentistry, oral surgery, orthopedics, sport medicine, dermatology, and plastic and reconstructive surgery. Currently, different techniques to obtain these hemocomponents are available, each leading to preparations that may differ in platelet concentration, leukocyte and fibrin content. Among them, there is Plasma Rich in Growth Factors (PRGF), a preparation rich in platelets, easily obtained from a small volume of patient’s blood, which, being autologous, does not promote any immune reaction and infectious disease transmission. Additionally, leukocytes are excluded from PRGF, so as to avoid any pro-inflammatory effects. Despite the growing craze for platelet concentrates technology, the scientific literature reported controversial results regarding the beneficial effect of these preparations on tissue healing, especially for bone regeneration. Considering all this information, the aim of our research was to developed a study model in order to gain more information about the biological effect of a platelet concentrate on both soft and hard tissue regeneration. For this purpose, human adipose-derived stem cells (hASCs), human osteoblasts (hObs) and human dermal fibroblasts (hDFs) cultured in the presence of PRGF, were studied. Usually, cells used for human therapy are expanded in fetal bovine serum (FBS), but this way carries the risk of potential immunogenic residual bovine proteins exposure and possible contamination with infectious agents, likely generating immune responses in patients. Therefore, in this study, FBS was completely substituted by the platelet concentrate for the cells culture. Interestingly, PRGF never affected cell viability and, after a short adaptation period, both hASCs and hDFs grown in the presence of PRGF, increased their proliferation rate compared to standard culture condition, while hObs growth was evident only without any cell’s detachment. Furthermore, neither donor nor cell population effect on both cell viability and proliferation was observed. Regarding osteo-differentiation, alkaline phoshatase (ALP) activity was induced in both hASCs and hObs, with a peak at day 7 and 14, respectively. In parallel, in order to elucidate the clinical benefit of platelet concentrates in hard tissue repair, the combination of a bone graft substitute with PRGF in enhancing a bone defect regeneration performed in a preclinical animal model, was also investigated. In particular, bilateral circular critical lesions were created in the proximal tibia of six New Zealand rabbits, for a total of 12 defects, filled with the bone substitute alone or in association with PRGF. Untreated defects were also included as control. No side-effects have been observed during the 8 weeks follow-up. Both histological and histomorphometric analysis showed a non-significant difference between treatment groups in terms of bone density, even if the addition of PRGF to the bone substitute seemed to induce a higher percentage of newly formed bone. In conclusion, this in vitro and in vivo study provided the evidence of the beneficial effect of platelet concentrates in both soft and hard tissue regeneration, thus supporting the application of this technology in clinical regenerative therapies.

CARDIOVASCULAR PROTEOMICS AND MITRAL VALVE DISEASE IN DOGS: SEARCHING FOR A SEROLOGICAL BIOMARKER Myxomatous valve disease (MVD) is the most common heart valvular disease in humans in Europe and United States, and it’s the primary naturally occurring heart disease in dogs. MVD is pathologically identical in humans and dogs, suggesting a common pathogenesis in these species, and creating an increasing interest in the canine MVD as a model for the human medicine. Nowadays, MVD is one of the most studied CVDs, because of its high prevalence in the clinical practice. Thanks to the research efforts, great diagnostic and therapeutic progresses have been made in the last decades, and continuous improvement are in the making. In recent years, new technologies have been used by the cardiovascular medicine, and the advancement of proteomic techniques has improved the methods available for investigating CVDs. Proteomics is the large-scale study of proteins and its application to uncover the protein function and structure in normal or disease states in the cardiovascular field is called cardiovascular proteomics. Even if the use of cardiovascular biomarkers is widespread in both human and veterinary medicine, the application of cardiovascular proteomics to the MVD study has recently begun. In the present study we selected a cohort of private-owned dogs recruited from the Cardiologic Service of the Department of Veterinary Science and Public Health. The dogs were selected with precise inclusion and exclusion criteria. Two breeds were considered: Cirneco dell’Etna and Cavalier King Charles Spaniels. The Cirneco dell’Etna breed is a poorly diffuse Italian hunting breed whose CVDs prevalence has never been studied before, while the Cavalier King Charles Spaniel breed is the most studied breed for all the MVD researches, because of the high prevalence of MVD, the early onset and the strong scientifically proved hereditary component. The objective of the present study was to search one or more than one serological biomarkers in dogs affected by MVD, comparing blood samples from the healthy dogs of the groups with blood samples from the dogs affected by different stages of MVD. The proteomics results were then matched with the clinical and echocardiographic data obtained in the clinical trial of all the patients included in the study, to find a connection available in the clinical practice. 64% of the Cirneco dell’Etna dogs included in the study were affected by MVD, and all the dogs older than 6 years had echocardiographic signs of MVD. The proteomic analysis of the Cirneco samples gave the following results: the alpha-1-antytripsina (A1AT) was up-regulated in the patients affected by MVD, according to the severity of the pathology, while the complement C3 was down-regulated with the development of MVD, according to the stage of the disease. Among the Cavalier King Charles breed there was a high prevalence of MVD (63%) and a very low medium age of onset (4 years). The proteomic analysis of the Cavalier King Charles Spaniel samples produced the following results: the serum albumin was down-regulated, according to the severity of the pathology, while it was observed a strong up-regulation of some specific types of IgG and IgM, according to the severity of the pathology. Based on our study results, the Cirneco dell’Etna breed is a primitive hunting breed predisposed to the development of MVD, with an early onset of the pathology. All the CdE dogs older than 6 years should be therefore evaluated for MVD, and included in a screening program. We confirmed that Cavalier King Charles Spaniel is a breed with a high prevalence of MVD and an early onset too, as previously reported. The proteomic analysis conducted on our samples and correlated to the clinical results, indicate that the MVD is a pathology that is strictly connected to a chronic inflammation state. The up-regulation of A1AT, IgG, IgM, and the down-regulation of complement C3 and serum albumin are connected with an inflammatory status, that cause a depletion of the components of the complement system, an activation of the acute phase proteins and of the components of immunity response like IgG and IgM immunoglobulins. The hypothesis that MVD could be related to a chronic inflammation was already speculated in the last years, and, based on the present study results, we think that the analysis of the inflammatory mediators in MVD patients could be a great chance to uncover the pathogenic mechanism at the base of mitral valve disease.

Patients with acute coronary syndromes (ACS) are treated with dual antiplatelet therapy (DAPT), which includes aspirin, an inhibitor of thromboxane A2 (TxA2) production, and an antagonist of the P2Y12 receptor for ADP. Based on the recent observation that P2Y12 antagonists also inhibit the platelet production of TxA2, it has been suggested that patients with ACS might be safely treated with P2Y12 antagonists only. However, the observation that platelets congenitally deficient of P2Y12 synthesize normal amounts of TxA2 contrasts the results obtained with P2Y12 antagonists. To test whether the reported inhibitory effect of P2Y12 antagonists on TxA2 production is due to off-target effects, or secondary to inhibition of platelet aggregation (PA). Serum TxB2 was measured in 2 patients with inherited P2Y12 deficiency and 7 healthy subjects in presence/absence of increasing concentrations of P2Y12 antagonists added in vitro, and in 20 patients treated with 10 mg/d prasugrel (P2Y12 antagonist) or placebo for 14 days in a randomized, double-blind, cross-over study. TxB2 levels were also measured after stimulation of citrate-PRP by collagen (0,5µg/mL) or arachidonic acid (1mM) in an aggregometer in presence/absence of P2Y12 antagonists, under stirring and non-stirring conditions (PA does not occur without stirring). P2Y12 antagonists did not decrease serum TxB2 levels both in vitro and ex vivo (prasugrel-treated patients). They partially inhibited TxB2 production under stirring, but not under non-stirring conditions. In conclusion, P2Y12 antagonists do not inhibit the platelet TxA2 production; therefore, there is no pharmacological evidence that aspirin should be withheld in patients with ACS.

Unrelenting advances in the field of nanoscience are fostering the progress in diverse research fields, ranging from light-emitting to medicine and diagnostics, from energy conversion to communication technologies. Besides representing the most paradigmatic example of nanoscience, semiconductor quantum dots (QDs) avowedly brought revolutions in many of the research fields mentioned above. Nowadays, some QDs-based devices and applications reported efficiencies almost as good as current state-of-the-art technologies. The founding concept of QDs is the application of quantum confinement effects on excitons, i.e., the main players of optical properties in bulk semiconductors. Among the wealth of ensuing properties, the size- and shape- tunability of the electronic excitations and increased coupling with light field aroused much interest. Also, the colloidal approach endows QDs with high processability and low cost, thereby encouraging their implementation in existing technologies and extending their impact to other fields. Howbeit, despite three decades of investigations, the bottom line has not been reached yet, and researchers are still delving deeper into the photophysics of these nanosystems. Though many of the low hanging fruit of QDs have been harvested, higher-lying ones seem to be even more succulent. This thesis deals with the quest for highly performing nanostructures, as a prerequisite for some high impact optoelectronic applications, e.g., QD-Lasers and QD-Solar Cells. Within this framework, the struggle against fast Auger recombinations and trapping of either hot carriers or cold excitons was addressed mainly by sophisticated core/shell technologies. Thus, the first part of the thesis reports how tuning different shell parameters (e.g., the smoothness of the interface potential, the height of the confining potential, and the interfacial strain) it is possible to exert control on these detrimental recombination processes. Though often disregarded, even the role of organic capping ligand is reconsidered in promoting the outcoupling of QDs excited states and addressing their interaction. Besides the useful and technologically relevant advice gathered within these studies, the primary inheritance of the first part is the comprehensive photophysical scenario, portrayed by a phenomenological model that successfully describes many aspects of the exciton dynamics in QDs. This amount of knowledge was capitalized in the second part of this thesis, dealing with the quest for novel materials, potentially outpacing conventional CdSe-based QDs. Perovskite-based QDs reported promising results, whereas some pitfall in the conventional characterization of carbon-based QDs were discovered. The rationalization of both nature and dynamics of this materials is expected to expedite their development as alternative (and potentially superior) technologies concerning those studied in the first part.
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As equipes brasileiras de voleibol sempre tiveram os melhores levantadores, no cenário mundial, tais como: William, Jaqueline, Maurício, Fernanda Venturini, entre outros. Esta posição é considerada como a função mais importante. Isto nos leva a acreditar na importância das características físicas, técnicas, táticas e psicológicas de um levantador. Para o estudo foram entrevistados técnicos e atletas de equipes na categoria adulta (masculinas e femininas) participantes da Superliga atual campeonato nacional. Como método utilizou-se uma entrevista sem iestruturada com estes técnicos e atletas. Para análise qualitativa foi aplicada a metodologia do DSC (Discurso do Sujeito Coletivo). Trata-se de uma técnica de pesquisa qualitativa que está sendo desenvolvida no campo da saúde desde 1997, mas que tem utilidade em todo tipo de abordagem da temática que envolva o pensamento, o sentimento, as crenças, as atitudes, os valores, as representações sociais, expressados sob a forma de discursos verbais. O procedimento da categorização das respostas (O Pensamento Coletivo como Soma Qualitativa) foi aplicado nessa fase. Consistiu em agrupar as respostas de sentido equivalente \"assim como sua força - freqüência com que uma dada Idéia Central apareceu no campo pesquisado. Das diversas questões abordadas nesta entrevista, evidenciou-se para esta pesquisa a seguinte: \"Para você, quais seriam as variáveis que um levantador deve ter para ser um atleta de alto rendimento e chegar a uma seleção nacional?\"
Brazilian volleyball teams always had the best setters worldwide, such as William, Mauricio, Jaqueline, Fernanda Venturini among others. This position is considered the most important in the team, reason why we may conclude that physical, technical, tactical and psychological characteristics are very important in a setter. The sample was composed of adult teams (male and female) coaches and athletes from \"SUPERLlGA\" - Brazilian National Championship. The methodology used was a semi structured interview. For the qualitative analysis was used the CSS (Collective Subject Speech) , a qualitative technical research that is being developed in the Health Area since 1997 and that is helpful for any study involving thinking, altitude, feeling, social representation, beliefs, expressed by verbal speech. The answers computing (collective thinking as qualitative addition) was applied at this time, and consists of joining the same sense answers as well as the incidence of the central idea occurrence in the study. Among all the questions made in this presentation one carne up: 11 What would be the most important characteristics (physical, technical, tactical and psychological) that a setter must have to acquire a high performance profile and reach a National team? \"

The herein reported thesis finds its roots in the concepts of sustainability and circular economy and has as purpose the valorisation of agro- and fishery-waste for the production of added-value products. In particular, different types of waste biomass were used as starting material for the synthesis of carbon dots (CDs) used as photocatalysts, for the obtainment of biopolymers, namely collagen and chitin, and for the extraction of valuable compounds using supercritical CO2. The investigation started studying the dependence of structure and photocatalytic behaviour of CDs on the carbon source. Six different types of carbon nanoparticles were synthesized and compared from the morphological and optical point of view revealing a strong correlation of their properties with the starting material and the synthetic method. The citric acid-derived CDs were then employed to photoactivate the atom transfer radical polymerization of a methacrylate leading to the obtainment of the polymer in high conversions (89%) and narrow dispersity (1.4). The study on CDs proceeded then with the employment of more complex starting material, namely fish scales. Naturally nitrogen doped carbon dots were successfully synthesized starting from bass scales, fully characterized and employed as photocatalyst for the reduction of methyl viologen. The bass-CDs were compared with classic citric acid-derived CDs resulting in higher initial photoreduction rate (7.5·10−8 M·s−1 vs 4.9·10−8 M·s−1). The fish-derived nanoparticles were then applied as photocatalyst for the continuous flow degradation of azo dyes highlighting a quantitative degradation of five model dyes in only 2 min. This study was conducted during a research period abroad (September 2021-April 2022) in the laboratories of the Université de Liège under the supervision of Prof. Jean-Christophe Monbaliu. The upgrading of fishery waste was then developed more during the second part of this thesis where the possibility to extract biopolymers from fish scales or crab carapaces was explored. After a review study on the production of UV-shields using biopolymers, an investigation on the preparation of UV-blocking films from mullet scales-derived gelatin and bass-CDs was conducted. The obtained materials were fully characterized from the morphological, mechanical and optical point of view; by adding just 5% of CDs the films blocked almost 70% of the UV radiation with negligible change in opacity and in transparency. A further study on the possibility to obtain chitin from crab shells using a one-step protocol using ionic liquids was then conducted. All characterization data confirmed that in the conditions tested, ammonium formate prepared in situ seems to be a promising candidate for one-pot chitin pulping process, allowing quantitative isolation, high purity and a high degree of acetylation (DA > 90%). To have a complete overview on the possibility to exploit waste biomass for the production of added-value products, an investigation on the valorisation of agro-waste was then conducted. In this frame, a supercritical CO2 extraction method was developed to obtain natural preservatives from hops and jimsonweed. scCO2 extracts were compared to ethanolic ones: the supercritical fluid led to the selective extraction of volatile compounds resulting in a better recovery for preservatives. The scCO2 technique was then applied to obtain fatty acids for cosmetic formulations from fruit pomace, resulting again more selective when compared to classical organic solvents.
The herein reported thesis finds its roots in the concepts of sustainability and circular economy and has as purpose the valorisation of agro- and fishery-waste for the production of added-value products. In particular, different types of waste biomass were used as starting material for the synthesis of carbon dots (CDs) used as photocatalysts, for the obtainment of biopolymers, namely collagen and chitin, and for the extraction of valuable compounds using supercritical CO2. The investigation started studying the dependence of structure and photocatalytic behaviour of CDs on the carbon source. Six different types of carbon nanoparticles were synthesized and compared from the morphological and optical point of view revealing a strong correlation of their properties with the starting material and the synthetic method. The citric acid-derived CDs were then employed to photoactivate the atom transfer radical polymerization of a methacrylate leading to the obtainment of the polymer in high conversions (89%) and narrow dispersity (1.4). The study on CDs proceeded then with the employment of more complex starting material, namely fish scales. Naturally nitrogen doped carbon dots were successfully synthesized starting from bass scales, fully characterized and employed as photocatalyst for the reduction of methyl viologen. The bass-CDs were compared with classic citric acid-derived CDs resulting in higher initial photoreduction rate (7.5·10−8 M·s−1 vs 4.9·10−8 M·s−1). The fish-derived nanoparticles were then applied as photocatalyst for the continuous flow degradation of azo dyes highlighting a quantitative degradation of five model dyes in only 2 min. This study was conducted during a research period abroad (September 2021-April 2022) in the laboratories of the Université de Liège under the supervision of Prof. Jean-Christophe Monbaliu. The upgrading of fishery waste was then developed more during the second part of this thesis where the possibility to extract biopolymers from fish scales or crab carapaces was explored. After a review study on the production of UV-shields using biopolymers, an investigation on the preparation of UV-blocking films from mullet scales-derived gelatin and bass-CDs was conducted. The obtained materials were fully characterized from the morphological, mechanical and optical point of view; by adding just 5% of CDs the films blocked almost 70% of the UV radiation with negligible change in opacity and in transparency. A further study on the possibility to obtain chitin from crab shells using a one-step protocol using ionic liquids was then conducted. All characterization data confirmed that in the conditions tested, ammonium formate prepared in situ seems to be a promising candidate for one-pot chitin pulping process, allowing quantitative isolation, high purity and a high degree of acetylation (DA > 90%). To have a complete overview on the possibility to exploit waste biomass for the production of added-value products, an investigation on the valorisation of agro-waste was then conducted. In this frame, a supercritical CO2 extraction method was developed to obtain natural preservatives from hops and jimsonweed. scCO2 extracts were compared to ethanolic ones: the supercritical fluid led to the selective extraction of volatile compounds resulting in a better recovery for preservatives. The scCO2 technique was then applied to obtain fatty acids for cosmetic formulations from fruit pomace, resulting again more selective when compared to classical organic solvents.

Animals are considered sentient beings by law and the notion of animal welfare, both physical and psychological, is well-established (“Five Freedoms”). As a result, regulation on animal testing puts researchers through highly regulated and restrained protocols (to reduce the number of animals included in in vivo experiments and replace them with alternative procedures whenever possible. This project revolves around the study of the bone tissue in different species, grounding on the idea of cadaver “recycle” applied to bone-related research. The bone, in fact, is particularly easy to study in cadavers since it suffers the early consequences of post-mortal decay to a lesser extent compared to other tissues. The bone is a highly specialized connective tissue characterized by a mineralized extracellular matrix, specifically designed for structural, mechanical and metabolic functions. Despite general shared features, interspecific differences exist in bone microscopic organisation, which have not yet been completely elucidated. Because of its peculiarities, a comprehensive evaluation of the bone includes several different techniques, such as histology, densitometry and biomechanics. Histologic techniques are particularly common, especially when it is necessary to evaluate tissue biocompatibility and integration in animal models of bone healing and regeneration. However, due to highly mineralized nature of its matrix, several problems arise during bone processing, especially during decalcification (the removal of mineral from the bone while preserving all the essential microscopic elements and tissue antigenicity), a process that is yet far from being standardized, but is necessary to prepare paraffin-embedded specimens. The first two aims of this project were the definition of species-specific and site-specific guidelines for bone specimens’ decalcification and the characterization of swine and ovine bone tissue with a multidisciplinary approach. Basic protocols for trabecular bone decalcification in different species with different solutions were provided, showing that excellent morphologic results can be achieved with solution containing strong acids in rodents, sheep and pig. As a general rule, hydrochloric acid solution proved to be too aggressive, as previously reported. On the other hand, the combined use of hydrochloric acid and formic acid proved to be a very good compromise, shortening experimental time and preserving morphology. Finally, a combination of citric acid formic acid provided excellent morphology, but at the expenses of decalcification time. Collagen I was preserved in swine, canine and ovine samples that proved to have good/excellent morphology, while it wasn’t preserved in rats or mice, despite their excellent morphology. A multidisciplinary approach to the study of the bone in pigs and sheep was developed, evaluating several different parameters with different techniques on the same samples, also providing standard reference parameters about swine and ovine bone histology, densitometry and biomechanics, to be compared with future research results, to deepen the knowledge on the bone tissue and facilitate biomedical research on animal models. The histogenesis of bone can be divided in two modalities: endochondral and intramembranous, which share a common sequence of events. Bone growth starts during the fetal stage, and then continues after birth together with the growth of the entire individual. Special structures, called cartilage canals, are involved in the process of endochondral ossification, with nutrients supply to the growing cartilage and waste products elimination being their primary functions. Cartilage canals have been described in several species, but little information is known about them in dogs. The development of a newborn into an adult implies dramatic changes in both body size and shape. The dog, in particular, exhibits the highest intraspecific differences in body shape and size among mammals, and this might be reflected by different growth patterns. This is especially true when the skull is considered, as it is the most variable body part among different breeds. Little information is known about skull development in dogs in relation to their adult craniometric category (brachycephalic, mesaticephalic, dolychocephalic). Due to the extreme plasticity of young bones, moreover, skeletal development could potentially be influenced by the presence of orthopaedic diseases, which could modify the normal shape and size of the future adult bones. The last three aims of this project were the study of endochondral ossification in long bones, with particular emphasis on the role role of cartilage canals; the study of the effects of pathology (quadriceps contracture) on hindlimb development; and the study of skull development and its compared features in brachycephalic, mesaticephalic and dolychocephalic breeds. New insights on cartilage canals in small-sized dogs were provided, which confirmed their involvement in bone extracellular matrix production in dogs, as in other species, laying the basis for more extensive studies on the delicate mechanism of the formation of ossification centres and the regulation of ossification. The abnormalities induced by quadriceps contracture on hind limb skeletal development, mainly consisted of volume reduction and abnormal shape of several centres, were described in a litter of Dobermann. These alterations that should be carefully considered when evaluating puppies affected by the disease to plan a therapy. Finally, novel contribution in canine craniometry and skull development were provided, documenting morphometric differences among dolichocephalic, mesaticephalic and brachycephalic purebred puppies in the early neonatal period through a radiographic and anatomic study. Cadavers invariably proved to be an invaluable source of research material, providing a valid alternative to in vivo experiments and contributing to the reduction of the use of experimental animals.

La terapia delle lesioni spinali croniche dell'uomo rappresenta attualmente uno dei maggiori campi di interesse della ricerca scientifica. Tra le strategie sperimentali di ultima elaborazione, il trapianto di cellule staminali sta dimostrando notevoli potenzialita'  per la cura di molte patologie del SNC. I modelli animali principalmente utilizzati per gli studi sperimentali sono i ratti, ma le dimensioni del midollo spinale in questa specie sono estremamente diverse da quelle dell'uomo, e le condizioni di laboratorio in cui i protocolli sperimentali vengono applicati sono molto differenti dalle condizioni accidentali delle lesioni umane. Il trauma spinale e' un'evenienza piuttosto frequente nel cane, ma molto spesso la terapia non viene effettuata precocemente o e' inefficace, cosi' da portare ad una condizione di paraplegia cronica. In questa specie l'eziopatogenesi delle lesioni e le dimensioni del midollo sono simili a quelle dell'uomo. Per tali ragioni il cane rappresenta un buon modello per lo studio delle terapie sperimentali nelle lesioni spinali croniche. Lo scopo del presente lavoro e' quello di verificare se l'impianto autologo di cellule staminali adulte di natura olfattoria sia in grado di ripristinare la deambulazione volontaria in cani paraplegici cronici. Sono stati inclusi nello studio 6 soggetti sottoposti alla nostra attenzione tra gennaio 2004 e dicembre 2005 per l'insorgenza di un grave trauma spinale compreso tra le vertebre T4 e L3. Trascorsi almeno due mesi dall'insorgenza della paraplegia, in ciascun cane e' stato effettuato il prelievo bioptico della mucosa nasale. Il campione e' stato quindi inviato nel laboratorio di riferimento, dove le cellule staminali sono state isolate e allestite su un apposito supporto. Nei mesi successivi tutti i 6 pazienti sono stati sottoposti all'impianto delle cellule autologhe, dopo esposizione chirurgica del segmento midollare leso e asportazione del tessuto cicatriziale. La fase post-operatoria e' stata dedicata all'esecuzione di intensi protocolli fisioterapici e di indagini cliniche e strumentali, finalizzate alla valutazione del recupero neurologico. I risultati ottenuti hanno evidenziato la presenza di un parziale ripristino delle capacita' deambulatorie in 3 soggetti, mentre i tracciati elettrofisiologici relativi ai Potenziali Evocati Somato-Sensoriali (PESS) non hanno subito alcuna variazione nel periodo successivo all'impianto. In un soggetto, deceduto spontaneamente per una patologia indipendente dalla condizione neurologica, e' stato possibile sottoporre il midollo spinale a valutazioni di tipo istologico e immunoistochimico. Queste hanno evidenziato la presenza, nel tratto midollare sede dell'impianto, di gruppi di neuroni-fibre dispersi all'interno di abbondante tessuto fibroso. Pur considerando l'esiguita' del numero dei soggetti inclusi nella ricerca, e in attesa di poter sottoporre ad indagini istologiche il midollo spinale di 5 di loro, si puo' concludere che l'impianto di cellule staminali di origine olfattoria nel midollo spinale di cani paraplegici cronici puo' presentare notevoli potenzialita' terapeutiche. The treatment of chronic spinal cord injuries in humans is, today, one of the most important fields of scientific research. Among the experimental strategies recently developed, stem cell transplants are demonstrating great potentiality for the cure of many central nervous system (CNS) pathologies. The most common animal models used in experimental studies are rats but the dimensions of the spinal cord of this species are quite different from those of the human, and the laboratory conditions in which the experimental protocols are applied do not resemble the accidental conditions of human injuries. Spinal cord injuries are relatively frequent in dogs but very often the therapy is not applied promptly or is not efficacious, and the result is a condition of chronic paraplegia. The etiopathogenesis of the injuries and the dimensions of the spine of the dog are similar to those of the human. For these reasons, the dog represents a good model for studying experimental treatment of chronic spinal cord injuries. The aim of this study is to verify whether the autologous implant of adult stem cells obtained from nasal olfactory mucosa (NOM) is able to restore voluntary ambulation in dogs with chronic paraplegia. This study concerns six subjects that we cared for between January 2004 and December 2005 after the onset of a serious spinal trauma in the area between the vertebrae T4 and L3. At least two months after the onset of paraplegia, a bioptic sample was taken from the nasal mucosa of each dog. The sample was sent to the reference laboratory where the stem cells were isolated and prepared on an appropriate scaffold. During the following months all six patients were subjected to the implant of autologous cells, after the surgical exposure of the injured segment of the spinal cord and the removal of scar tissue. The post-surgical phase was dedicated to intensive physiotherapy protocols and clinical and instrumental investigations, with the aim of evaluating the neurological recovery. The results obtained demonstrated the presence of a partial restoration of the ambulatory capacity in three subjects, while the electrophysiological pathways relative to the Somatosensory Evoked Potential (SSEPs) did not undergo any variation in the period following the implant. In the case of one dog, that died spontaneously from a pathology not connected with the neurological condition, it has been possible to carry out histological and immununohistochemical evaluations of the spinal cord. These have demonstrated the presence of groups of neuron-fibres dispersed inside abundant fibrous tissue in the area of the implant. While taking into consideration the small number of subjects included in the research and in anticipation of being able to subject the spinal cords of the other five dogs to histological investigations, we can nevertheless conclude that the implant of the stem cells of olfactory origin in the spinal cord of dogs with chronic paraplegia can offer outstanding therapeutic potential.

In this PhD thesis work I investigated the expression modulation of the major adhesion factors in Escherichia coli; in particular I focused on the role of GGDEF and EAL proteins, on their modulation in E. coli biofilm formation in response to environmental signals and on regulation of curli fibers, cellulose and poly-N-acetylglucosamine (PNAG), the most important biofilm determinants in E. coli. E. coli is an Enterobacterium, normally living inside the mammalian gut, at temperature of 37° C and in relatively nutrient-rich environment. Once outside the host, bacteria usually face much lower temperatures (< 30°C) and a nutrient-limiting environment. The biofilm determinants studied in this thesis are all expressed in response to environmental conditions such as low temperature, low osmolarity and starvation, suggesting that E. coli bacteria switch to a biofilm mode of growth as part of their adaptation to the natural environment. In response to reduction in growth rates, E. coli seems to canalize its energy consumption into production of extracellular features such as curli or exopolysaccharides. Biofilms can be thus considered as a “resistance form” of growth able to withstand stress conditions more efficiently than cells living in a planktonic mode of growth. The CsgD protein is the master regulator of E. coli biofilm formation. It is a transcriptional factor necessary for curli genes transcription and, through the AdrA protein, for cellulose biosynthesis. Gene regulation by CsgD is tightly connected to production and sensing of cyclic di-GMP, a bacterial second messenger involved in various cellular processes, including biosynthesis of extracellular polysaccharides (Simm et al., 2004), biofilm formation (Hickman et al., 2005), and virulence (Pratt et al., 2007; Tischler and Camilli, 2005), as well as morphological and physiological differentiation (Paul et al., 2004). The CsgD-dependent adrA gene, involved in cellulose biosynthesis (Zogaj et al., 2001), encodes a cyclic di-GMP synthase (Simm et al., 2004). CsgD can also activate yoaD, whose gene product is a cyclic di-GMP phosphodiesterase, suggesting that CsgD is directly involved in feedback regulation of cyclic di-GMP intracellular levels and of cellulose biosynthesis (Brombacher et al., 2006). CsgD is also able to activate the iraP gene: IraP acts as a stabilization factor for the σs protein, an alternative sigma factor of RNA polymerase which directs transcription of genes involved in adaptation to slow growth and to cellular stresses. Here I showed that CsgD transcription activation of the iraP gene does result in a significant increase of σs intracellular concentration by positively affecting σs protein stability, thus leading to altered expression of σs-dependent genes. CsgD-mediated increase of σs cellular concentrations via the iraP gene would trigger an autoactivation loop leading to an increased production of CsgD-dependent adhesion determinants such as curli fibers and cellulose. This autoregulatory circuitry might be further fueled by σs-dependent induction of genes encoding di-guanylate cyclases, i.e., proteins able to synthesize the second messenger di-cyclic- GMP, which, in turn, can positively affect csg gene expression (Kader et al., 2006; Weber et al., 2002). The yddV-dos operon is the most expressed among c-di-GMP-related genes showing dependence on σs (Weber et al., 2006; Sommerfeldt et al., 2009). It encodes, respectively, a protein with DGC activity and a PDE that can degrade c-di- GMP to pGpG. Both Dos and YddV are heme-binding oxygen sensors, and interact to form a stable protein complex (Tuckerman et al., 2009). Although it has been reported that YddV overexpression can stimulate biofilm formation (Mendez-Ortiz et al., 2006), the targets of yddV-dependent biofilm induction had not yet been identified. Here I showed that YddV acts modulating curli and PNAG expression. Control of curli production by yddV-dos takes place at the level of transcription regulation of the csgBAC operon, encoding curli structural subunits, and is mediated by the DGC and PDE activities of YddV and Dos. In contrast, the YddV–Dos protein complex does not strongly influence csgDEFG expression, nor does it affect the expression of the CsgD-dependent adrA gene, encoding a positive effector for cellulose biosynthesis. Regarding PNAG production, we showed that YddV is able to prevent degradation of pgaABCD transcript in the MG1655csrA background, thus suggesting that a DGC might regulate gene expression by affecting mRNA stability in E. coli. YddV regulation of pgaABCD operon in a wild type contest is still controversial: pgaABCD genes are expressed at low levels in MG1655 (the standard laboratory strain of E. coli) and their mRNA half-life is lower than two minutes regardless of the growth conditions tested; thus, possible effects of yddV inactivation on destabilization of the pga transcript are not easy to evaluate in the wt contest. In the last part of my thesis I tried to characterize a biofilm-forming mutant of E. coli, able to express pgaABCD genes at high levels. Even if initial data suggested that a mutation in the csrA gene could be responsible for pga mRNA stabilization in this mutant, actual the mutation leading to the adhesive phenotype and to PNAG production is outside the csrA gene and is still unknown. Moreover my data suggest a connection between pga expression and iron regulation in E. coli strains: it is conceivable that pgaABCD expression and consequent biofilm formation and the adherent phenotype depends on concerted production of different determinants, whose expression is also affected by iron concentration. Thus, my research highlighted that biofilm production is the result of coordinated expression of different adhesion determinants, whose regulation is complex and not fully understood. In particular, the precise extent and the molecular mechanism of c-di-GMP adhesion factors regulation remains to be largely identified and represents an exciting challenge for future research in the biofilm field.

L'esame clinico unitamente agli esami di diagnostica collaterale di laboratorio e di diagnostica per immagine risultano spesso efficaci nel sospettare la presenza di una patologia epatica, ma solo raramente permettono una diagnosi definitiva, per la quale a volte e' necessario ricorrere a tecniche piu' invasive quali l'esame citologico e/o istologico. Inoltre, analogamente a quanto accade per la medicina umana, anche in ambito veterinario si e' cercato di verificare l'effettiva utilita' diagnostica di alcuni marcatori tumorali, nello svelare precocemente i soggetti affetti da neoplasie epatiche primarie o metastatiche. Gli obiettivi che il presente lavoro si e' prefisso sono stati i seguenti: · valutare la sensibilita' e la specificita' di alcuni parametri di biochimica clinica (in particolare di ALT, AST, ALP, γGT e GLDH) e verificarne il grado di correlazione in corso delle principali patologie a livello epatico; · stimare l'accuratezza diagnostica di tecniche poco invasive, quali la FNAB (fine-Needle Aspiration Biopsy) eco-guidata nella diagnosi precoce di alterazioni epatiche; · stabilire l'effettiva utilita'  diagnostica in citopatologia di alcune colorazioni speciali quali PAS ed OIL RED-O nel differenziare l'epatopatia vacuolare steroido-indotta da quella da accumulo di lipidi; · cercare di valutare il ruolo diagnostico e prognostico di alcuni marcatori tumorali, quali l'AFP, il CEA ed il CA 19,9, quali indicatori di patologie neoplastiche a carico del fegato. A tale scopo, sono stati analizzati i risultati provenienti da 38 pazienti, tutti appartenenti alla specie canina, che presentavano, sulla scorta dell'esame clinico diretto e/o alla valutazione sierica del profilo epatico sopra elencato, alterazioni compatibili con compromissione epatica. Per meglio allestire una valutazione statistica, e' stato inoltre preso in considerazione un gruppo di 10 cani sani, utilizzati come controllo. I cani oggetto del presente lavoro sono stati sottoposti a prelievo epatico con tecnica di ago-infissione per la valutazione citologica e mediante ago da biopsia per quella istologia. Entrambe le procedure sono state eseguite sotto guida ecografica e con accesso percutaneo. Inoltre su siero si sono eseguite le determinazioni relative ai marcatori tumorali di AFP, CEA e CA 19,9. I risultati relativi alle valutazioni ematobiochimiche hanno mostrato i seguenti valori di: AST ed ALT elevati rispetto ai valori di riferimento rispettivamente in 24 e 15 soggetti, 33 pazienti hanno mostrato aumenti della fosfatasi alcalina ALP, in 30 la γGT ha presentato alterazioni rispetto al range fisiologico. Aumenti dei livelli sierici del GLDH si sono verificati in 25 pazienti. La valutazione citologica ha evidenziato un quadro compatibile con diagnosi di fegato normale in 3 pazienti (Prev. app. 7,89%); patologia infiammatoria in 5 (Prev. app. 13,16%); epatopatia degenerativa vacuolare in 18 (Prev. app. 47,37%); accumulo di pigmento in 17 soggetti (Prev. app. 44,74%); iperplasia in 5 (Prev. app. 13,16%); neoplasia in 11 (Prev. app. 28,95%). Molti dei cani oggetto di tale studio (17/38) hanno presentato, alla valutazione cito-morfologica epatica, un quadro compatibile con piu' di una delle categorie diagnostiche considerate. Dei 18 campioni con diagnosi di epatopatia degenerativa vacuolare, 9 hanno mostrato una netta positivita' alla colorazione PAS e i restanti 9 sono risultati positivi alla OIL RED-O, con valori di sensibilita' e specificita' pari al 100%, rispetto alla valutazione istologica. L'esito dell'esame istologico ha fornito i seguenti risultati: in 3 soggetti il quadro istologico era risultato compatibile con fegato normale (Prev. reale 7,89%); la patologia infiammatoria, si e' riscontrata in 14 cani (Prev. reale 36,84%) e la epatopatia degenerativa vacuolare in 23 pazienti (Prev. reale 60,53%). In 16 soggetti si e' evidenziato accumulo di pigmento (Prev. reale 42,11%); in 4 iperplasia nodulare (Prev. reale 10,53%), mentre in 9 cani l'istologia ha diagnosticato neoplasia primaria o metastatica (Prev. reale 23,68%). Anche in questo caso in 23 soggetti coesistevano piu' categorie diagnostiche. Valori di AFP al di sopra del limite di riferimento, considerato per la specie canina, pari a minus 70 ng/ml, si sono verificati in 13 cani in 5 dei quali major250 ng/ml, mentre nei restanti 8 casi i valori della AFP erano inferiori a 250 ng/ml. Sia il CEA che il CA 19,9 sono risultati non dosabili in nessuno dei pazienti oggetto del nostro studio con valori inferiori a 0,5 ng/ml e minus 1 U/ml. Nei campioni compatibili con patologia infiammatoria, accumulo di pigmento e neoplasia tutti i parametri di biochimica clinica hanno mostrato una differenza significativa ( minus 0,05) tra i due gruppi considerati. Nei soggetti con fegato normale, epatopatia degenerativa vacuolare ed iperplasia gli unici parametri di biochimica clinica che hanno mostrato differenze significative ( minus 0,05) tra i due gruppi sono stati la ALP e la GLDH. Dai risultati ottenuti confrontando l'affidabilita' dell'esame citopatologico con quello istopatologico, emerge come la sensibilita' dell'esame citologico si e' attestata su valori variabili compresi tra il 35,71% (in corso di lesioni infiammatorie) ed il 100%, (fegato normale, accumulo di pigmento e condizione neoplastica). Nei campioni con diagnosi istologica di epatopatia degenerativa vacuolare la sensibilita' della citologia e' risultata pari al 78,26%. La sensibilita' e la specificita' delle colorazioni PAS ed OIL RED-O nel differenziare le lesioni steroido-indotte da quelle causate dall'accumulo di lipidi sono risultate entrambe pari al 100%. La specificita' dell'esame citologico ha mostrato valori variabili dal 91,18% (iperplasia epatocellulare) al 100% (fegato normale, condizione infiammatoria e epatopatia degenerativa vacuolare). I risultati ottenuti dalla valutazione del profilo ematico ci portano a considerare come, analogamente a quanto riportato in letteratura, i parametri biochimici da noi considerati, non hanno mostrato alcuna specificita' nel permettere di differenziare un evento neoplastico da una condizione infiammatoria o degenerativa a carico del fegato. La frequente presenza in uno stesso campione di pia'¹ tipi di lesioni per le quali il test citologico mostra sensibilita' e specificita' discordanti e la bassa sensibilita' riscontrata per maggior parte delle categorie diagnostiche, esclude l'utilizzo della metodica citologica quale valida sostituta a quella istologica. L'elevata sensibilita' e specificita' dimostrate dalle colorazioni PAS ed OIL RED-O nella corretta interpretazione delle strutture vacuolari intracitoplasmatiche, ne suggeriscono l'impiego nella pratica citologica quale valido ausilio diagnostico nel differenziare i casi con epatopatia steroido-indotta da quelli con lipidosi. Contrariamente a quanto descritto in letteratura, i risultati estrapolati dalla valutazione dell'AFP nei soggetti in esame, hanno mostrato i limiti di tale determinazione nel discriminare i soggetti con neoplasia epatica da quelli con epatite. I risultati ottenuti dalle valutazioni di CEA e di CA 19,9 eseguiti con metodiche routinariamente utilizzate nella diagnostica oncologica umana, ne suggeriscono la non applicabilita'  in medicina veterinaria.

Canine Leishmaniasis is a zoonotic disease that is spreading over mediterranean basin. Despite the pathogenesis of canine leishmaniasis has been investigated through several studies, some aspect of the host-parasite interaction that lead to overt diseases and typical inflammatory lesions have not yet been understood. Among these, the oxidant-anti-oxidant mechanisms that occur in dogs with the clinical disease are particularly interesting since on one hand they can provide to us useful information on the pathogenesis of the disease, which can also be a model for investigating immune-mediated chronic inflammatory conditions occurring also in other species on the other hand the identification of changes in molecules involved in oxidative stress may serve as a potential target for ancillary treatments or as markers of disease severity and progression. Similarly, most of the researches done until now have been focused on renal damage but only a few studies were focused on markers of tubular damage, despite the development of tubulo-interstitial nephritis is associated with more advanced stages of the disease. As regards inflammatory/oxidative responses, our studies confirmed the presence of oxidative phenomena associated with the inflammatory profiles of infected dogs and suggested that reactive oxygen species released by inflammatory cells are not the only oxidative mediators involved in this disease. The serial analysis of markers of inflammation or oxidation defined the possible role of these markers as prognostic factors, especially after treatment. More specifically, antioxidant molecules such as PON1 and HDL may play a role as early markers of remission after treatment since they quickly normalize in dogs that respond to treatments. As regards renal markers, we evidenced the possible role of GGT as a simple and inexpensive marker of tubular damage in dogs with leishmaniasis.

DOES SOUND MATTER? Studies of information encoding in short-term memory of signers and speakers. Summary The short-term memory (STM) span, which corresponds to the longest sequence of items correctly recalled in a specific order, represents a widely used measure of STM capacity. STM span is shorter with signs as compared to speech, a robust finding that has been documented in different languages and populations using a variety of experimental paradigms. Attempts to characterize the source of modality-specific variations in STM span have been primarily of two types. Some accounts have drawn attention to structural differences between signs and verbal stimuli, supposing that signs are more complex in their internal structure and require more processing load for simultaneous integration of different features (movement, orientation, location, handshape) and longer articulatory duration. While a few results lent support to accounts assuming structural differences, findings showing that shorter STM spans persisted even when signs were carefully matched to verbal stimuli in duration, complexity, similarity etc., weaken accounts that identify structural differences as a primary cause of the disadvantage observed with signs. A second type of accounts hinges on the hypothesis that the reduced span is an effect of modality, stemming from greater STM capacity for encoding serial information in auditory STM as compared to visuo-spatial STM. The critical role these accounts assign to the encoding of temporal order information is justified by findings showing that when the task was free recall, without a requirement of a specific order, comparable spans appeared across modalities. The findings from free recall demonstrate comparable encoding of sign and speech, and working memory tests show comparable performance across modalities. Therefore the disadvantages for signs restricted to serial order recall lend support to a hypothesis linking the reduced STM capacity for signs to sequential order information. The present study aims to contribute to the investigation of the sequential order hypothesis that associates the reduced span of sign to the limitations of visuo-spatial STM processes in encoding temporal sequences. Research has shed light on the representation of serial order used in verbal STM, revealing that positions are encoded with respect to both start and end positions. The problem addressed in the present study is the source of the span reduction for ordered recall of signs, and a first question that arises is whether the same position scheme for order encoding in STM is used for signs and speech. We can explore this by identifying the scheme used to represent the position of items in a sequence in STM for signs and compare that to previous results from STM for speech. Here, we examined the perseverations that signers of Italian Sign Language produced in an immediate serial recall tasks. Perseverations were analysed to determine whether their occurrences reflected the encoding of serial positions with respect to both edges that characterizes verbal STM. In the core experiment of the present study we presented sequences of consonants of Italian sign language (LIS) alphabet to a group of 20 deaf students of Magarotto Institute in Padua, a secondary school where the communication among students and teachers is based on LIS. Sign sequences varying in length from 4 to 7 consonants were randomly presented on a computer screen at a rate of 1 second per sign. At the end of each sequence, participants were instructed to recall the letter signs in the same order. The length of the sequences was often purposely overspan to generate errors. We analysed the perseveration error pattern using the same technique as Fischer-Baum (2010). The results showed that, compared to hearing people, deaf participants demonstrated a reduced span, however, we found evidence for the same both-edges representation of position in STM for signed stimuli, suggesting that the same scheme is used to represent position in this task as is used in the STM for speech stimuli task. A next step was repeating the same experimental paradigm only using words and not letters as stimuli. Data was collected from 20 participants. However, in that case participants tended to produce responses of the same length, that made impossible to discern between different position encoding schemes. Those data were not enough to draw any definite conclusions from. A drastic extension of the subjects pool could have solved this problem, but at the moment it was not possible. There was, however, a possible bias: since the signers are literate, can read and write and possess strong skills of lip reading, there was a possibility that some phonological information could be involved in verbal material processing. To control for that, a study has been conducted involving two groups of participants: 20 signers (Magarotto institute students) and 15 speakers (University of Padua students). Two sets of experimental words were selected: similar in sign and similar in sound. Two matching control sets were balanced to experimental sets. The task was recalling sequences of 4 words, the 4 words were selected from one of the 4 sets. The hypothesis was that if the signers use phonological information, then words similar in sound would elicit more interference and would lead to weaker recall than a matching control group. However, the results demonstrate that signers show a strong effect of interference only in case of recall of words similar in sign, and no effect of similar sound, so we can conclude that there’s no phonological information involved in signers’ processing. An extended control study was also conducted to explore the contribution of the phonological loop in positional information encoding, since many theoretical explanations of the differences between STM of signers and speakers suggest that it’s the core structure for verbal information encoding for speakers. In this study the phonological loop was artificially blocked in speakers by means of adding articulatory suppression to the task of verbal stimuli memorizing and recall. However, the results of that study show that articulatory suppression leads participants to produce a greater rate of errors, but they use the same both-edges position encoding scheme, so we can conclude that the phonological loop doesn’t play a crucial role in serial order encoding scheme construction. We may have to search further for an intermediate problem of binding the elements of the input to serial position slots, that however exist in short term memory for signs as well as for spoken material, as it has been demonstrated by our studies.
Riassunto Lo span di memoria a breve termine (MBT), che corrisponde alla sequenza più lunga di elementi ricordati correttamente in un ordine specifico, rappresenta una misura della capacità di MBT ampiamente utilizzata. Lo span di MBT è più breve per il materiale in lingua dei segni rispetto al materiale in lingua parlata, è un risultato robusto che è stato confermato in diverse lingue e popolazioni utilizzando una varietà di paradigmi sperimentali. I tentativi di caratterizzare la fonte di differenze di span di MBT dovuti alla modalità di stimoli sono stati essenzialmente di due tipi. Alcuni hanno focalizzato la loro l'attenzione sulle differenze strutturali tra i segni e gli stimoli verbali, supponendo che i segni sono più complessi nella loro struttura interna e richiedono più carico di elaborazione per l'integrazione simultanea di diverse caratteristiche (movimento, orientamento, posizione, forma della mano) e la durata articolatoria più lunga. Mentre alcuni risultati mostrano evidenze a favore di questo approccio che assume differenze strutturali, ci sono altri risultati che dimostrano l’abbassamento di span di MBT anche quando i segni sono stati accuratamente abbinati e bilanciati rispetto agli stimoli verbali in durata, complessità, somiglianza ecc., e queste evidenze indeboliscono le ipotesi che assumono differenze strutturali come causa principale dello svantaggio osservato nello span per i segni. Un secondo tipo di approccio teorico si basa sull'ipotesi che lo span ridotto è proprio un effetto della modalità, derivante dalla maggiore capacità di MBT per codificare informazioni nell’ordine seriale nella modalità uditiva rispetto alla modalità visuo-spaziale. Il ruolo critico che questo secondo tipo di approcci assegnano alla codifica delle informazioni di ordine temporale è giustificato dai risultati che mostrano che quando il compito richiede di ricordare gli stimoli nell’ordine libero (free recall), senza l'obbligo di mantenere l’ordine di presentazione, lo span di MBT è comparabile tra le diverse modalità. I risultati di recall libero dimostrano capacità di codifica comparabili per segni e parlato, e anche i test di memoria di lavoro mostrano prestazioni confrontabili tra le diverse modalità. Quindi gli svantaggi per i segni sono limitati alla capacità di ricordare le sequenze di stimoli nell’ordine seriale, e questo rinforza l’ ipotesi che collega la capacità di MBT ridotta per i segni con la capacità di codificare le informazioni dell’ordine sequenziale. Il presente studio cerca di contribuire all’esplorazione dell'ipotesi dell’ordine sequenziale che associa lo span ridotto per i segni con dei limiti dei processi nella MBT visuospaziale nella codifica delle sequenze temporali. La ricerca ha messo in luce la questione delle rappresentazioni dell'ordine utilizzati nella MBT verbale, rivelando che in molti casi le posizioni sono codificati rispetto all’inizio e la fine della sequenza. La questione che ci poniamo nel presente studio è la fonte della riduzione di span per i segni, e, come primo passo, bisognerebbe capire se lo schema di rappresentazione della posizione per la codifica ordine in MBT è lo stesso per i segni e per il parlato. Possiamo esplorarlo identificando lo schema utilizzato per rappresentare la posizione degli elementi in una sequenza nella MBT per i segni e confrontandolo con i risultati precedenti ottenuti con la MBT per il parlato. Abbiamo esaminato gli errori di perseverazione che i parlanti della Lingua Italiana dei Segni producono in un compito di recall seriale ordinato che coinvolge i segni della lettera. Perseverazioni sono stati analizzati per determinare se i loro occorrenze riflettevano la codifica delle posizioni di serie rispetto a entrambi i bordi che caratterizza STM verbale. Nel principale esperimento del presente studio abbiamo presentato sequenze di consonanti di lingua dei segni italiana (LIS) ad un gruppo di 20 studenti sordi dell’Istituto Magarotto di Padova, una scuola specializzata in cui la comunicazione tra studenti e insegnanti è basata sulla LIS. Sequenze di segni di lunghezza da 4 a 7 consonanti sono stati presentati in ordine random sullo schermo del computer alla velocità di 1 segno per secondo. Alla fine di ogni sequenza, i partecipanti erano chiesti di ripetere i segni nell’ordine di presentazione. La lunghezza delle sequenze era spesso volutamente overspan in modo tale da generare errori. Abbiamo analizzato il pattern di errori di perseverazione con la stessa tecnica che ha usato Fischer-Baum (2010). I risultati hanno mostrato che, rispetto agli udenti, i partecipanti sordi hanno uno span di MBT ridotto, tuttavia, abbiamo trovato la conferma per la rappresentazione della posizione di un elemento della sequenza ancorata a due estremi della sequenza, suggerendo che lo stesso schema è utilizzato per rappresentare la posizione dei segni che per il materiale parlato. Un passo successivo era ripetere un esperimento con lo stesso paradigma sperimentale utilizzando parole e non le lettere come stimoli. I dati sono stati raccolti da 20 partecipanti. Tuttavia, in quel caso i partecipanti tendevano a produrre risposte della stessa lunghezza, che hanno reso impossibile distinguere tra diversi schemi di codifica di posizione. I dati ottenuti non sono stati sufficienti a trarre conclusioni definitive. Un aumento significativo della quantità di partecipanti avrebbe potuto risolvere questo problema, ma al momento non è stato possibile. C'era, però, un possibile bias: visto che i segnanti sordi sono comunque in grado di leggere e scrivere e hanno anche le competenze di lettura labiale, c'era una possibilità che alcune informazioni fonologiche potrebbero essere coinvolte nell’elaborazione del materiale verbale. Per controllare questo, abbiamo condotto uno studio con due gruppi di partecipanti: 20 segnanti (studenti dell'Istituto Magarotto) e 15 parlanti (studenti dell’Università di Padova). Abbiamo selezionato due gruppi di parole come stimoli sperimentali: parole simili in segno e simili il suono. Per il controllo abbiamo aggiunto due gruppi di parole corrispondenti, bilanciate alle parole sperimentali, ma dissimili tra di loro. Il compito era ricordare sequenze di 4 parole, dove la sequenza di 4 parole apparteneva a uno dei gruppi. Ci aspettavamo che se i segnanti usano informazioni fonologiche, allora le parole simili in suono avrebbero suscitato più interferenza portando alla prestazione peggiore che nel gruppo di controllo. Tuttavia, i risultati dimostrano che i segnanti dimostrano un forte effetto di interferenza solo in caso di parole simili in segno, e le parole simili in suono non elicitano nessun effetto. Allora possiamo concludere le informazioni fonologiche non sono coinvolte nell’elaborazione di stimoli segnati. Uno studio di controllo esteso è stato condotto per esplorare il contributo del loop fonologico nella codifica dell’informazione posizionale, visto che molte spiegazioni teoriche delle differenze tra MBT di segnanti e parlanti suggeriscono che è la struttura di base per la codifica dell’informazione verbale per i parlanti. In questo studio il loop fonologico dei parlanti è stato bloccato attraverso l'aggiunta di soppressione articolatoria al compito di memorizzazione di stimoli. I risultati dimostrano che la soppressione articolatoria porta i partecipanti a produrre un maggior numero di errori, ma, comunque, usano lo stesso schema di codifica di posizione seriale ancorata a due estremi della sequenza, quindi possiamo concludere che il loop fonologico non ha un ruolo importante nella costruzione dello schema di rappresentazione posizionale. We may have to search further for an intermediate problem of binding the elements of the input to serial position slots, that however exist in short term memory for signs as well as for spoken material, as it has been demonstrated by our studies. Quindi dovremmo cercare ulteriormente per un problema intermedio di collegamento di elementi dell’input ordinato alle posizioni rappresentate secondo lo schema ancorato a due estremi, lo schema, che comunque esiste anche nel caso dei segni, come abbiamo dimostrato, però si verifica più problematico il processo di posizionamento di elementi in questo schema.

The presence of liver fibrosis should be considered in order to define a correct therapeutical approach in liver diseases of the dog. Histologic evaluation of the liver is considered the gold standard for fibrosis assessment, but tru-cut liver biopsy, even if performed under ultrasound guidance, has some technical limitation and risks. In humans indirect biochemical markers, which reflect alteration in hepatic function, and direct biochemical markers, which reflect extracellular matrix turnover, have been identified. Objectives: to study the relationship between some non-invasive biochemical markers (AST/ALT ratio, APRI index and Hyaluronic acid HA) and hepatic fibrosis in the dog and to determine their value in the clinical practice. Methods: 32 blood samples (20 from dogs with chronic hepatitis diagnosed by liver biopsy and 12 from healty dogs) were collected for the study concerning AST/ALT ratio and APRI index. 18 blood samples (6 from pathologic dogs diagnosed by liver biopsy and 12 from healthy dogs) were tested for HA with ELISA immuno-assay. An histologic staging of liver flogosis and liver fibrosis was set up for this study and the results were compared with non-invasive indexes of liver fibrosis considered. Results: an AST/ALT ratio of 0.37 has 80% of sensibility and 91.67% of specificity for liver fibrosis in the dog (P< 0,01). No statistic correlation was detected between liver fibrosis and APRI index or HA. Conclusion: AST/ALT ratio in the dog seems to be correlated with liver fibrosis in the dog. Further studies should be considered to investigate the relation between AST/ALT ratio and the stage of fibrosis in the dog.

BACKGROUND - The urine test is probably the oldest laboratory examination of the history of medicine, and is still considered to be of undoubted utility and considered essential in the evaluation of diseases and urinary extraurinarie and monitoring of different metabolism. The chemical and physical examination of the urine in veterinary medicine, is still to be performed in ambulatory care in both clinical laboratories using a dry chemical method (dipstick), which allows the simultaneous determination of the following parameters: specific gravity, pH, leukocytes, nitrite, protein, glucose, ketone bodies, urobilinogen, bilirubin, blood and hemoglobin. Unfortunately, being born in human diagnostics, when used for the urine of dog and cat, do not provide reliable results for all tests. The systematic investigation of the physical-chemical parameters and determination of urinary analytes using standardized methods is a relatively recent acquisition, dating back to the 40s-50s. The legislation CLIA '88 (Clinical Laboratory Improvement Amendments) included examination of urine tests that can also run utside the laboratory, untrained operators, for its cheapness and speed of execution. However you must remember that this can be detrimental to the quality of results, which must be captured in an objective and especially knowing the factors that influence the sensitivity and specificity of the test. This makes it necessary to develop new methods that may also be more reliable and automated, to move from a semi-quantitative, such as that of the test strips, a quantitative determination of urinary chemical parameters. The new methods should also meet the various shortcomings of the dipstick in veterinary medicine, expanding, so the range of parameters commonly analyzed. An examination of urinary sediment, often overlooked and misunderstood, has a very useful diagnostic if correctly executed. In particular, Birch and Farley have reassessed the importance of the evaluation of haematuria in urinary sediment for the recognition of the anatomy of bleeding, in particular the study of the morphology of erythrocytes and the presence of acanthocyturia. The use of polarized light microscopy, phase contrast and improve the assessment of the elements of urinary sediment (in particular RBCs), the application of colors also allow extempuraneous staining evaluation of new elements of urinary sediment that may be indicative of renal disease. All this implies the need to introduce standard procedures in the collection, storage, preparation and analysis of the sample, such that the examination be reproducibily inside and outside the laboratory. AIM OF THE THESIS - The experimental work has focused on the evaluation of methods in liquid chemistry, recently introduced in human medicine for the quantitative determination of some urinary parameter (pH, specific gravity, glucose, bilirubin, nitrites, hemoglobin) in dogs and cats. These new methods will be applied to an automatic spectrophotometric commonly used for biochemical tests on serum. The results will be correlated with data obtained by dipstick and/or other urinary parameters. For the evaluation of the methods will be carried out repeatability and linearity tests on samples from both species. We want to emphasize that the automation of these methods is aimed to achieve several objectives, not influenced by the error of intra and inter-operator, then the improvement of analytical grade (for purposes of quality control laboratory) and the reliability of the data, no more qualitative or semi-quantitative, but quantitative. Furthermore, it highlights the economic importance of this method compared to the dry chemistry not only in terms of cost/reagent but also time-consuming. Another important step in the analysis of urine, often overlooked or misunderstood in laboratory medicine is the microscopic examination of urinary sediment. The application of standard procedures, new staining and multimodal reading of urinary sediment leads to a broadening of the diagnostic capabilities of this examination. MATERIALS AND METHODS - In the first study, the urinary specific gravity was determined by liquid chemistry method in samples of 84 dogs and 47 cats. The results were compared with data obtained by refractometer. The dipstick was excluded from this comparison to the previously documented unreliability in the species of interest. The data obtained showed that the determination of specific gravity in liquid chemistry has proved unenforceable in dogs and cats for the limited range of the method. In addition, when compared with the refractometric evaluation showed lack correlation. In an attempt to identify a substance can be determined and correlated with urinary specific gravity we have evaluated if there is a correlation between specific gravity and urinary creatinine (UCr) as has been demonstrated in humans.In the second part of the experimental work we have shown the first assessments of the mathematical model, and this correlation found between the two variables for the dog and cat. In the second, study the urinary pH was evaluated by liquid chemistry method, bench-top pH meter and dipstick. Data from urine samples of 205 dogs and 72 cats were compared. The presence of significant correlations between the three methods compared with each other, characterized by moderately high correlation coefficients in both species showed a close relationship between the methods considered. In the third study, we evaluated the kit for the quantitative determination of urinary glucose in liquid chemistry in urine samples from 207 dogs and 73 cats. The data will be compared with semi-quantitative assessments of the dipstick obtained on the same samples. The small number of samples positive for glycosuria has made statistical analysis inefficient but the quantitative method has proved especially useful in detecting low concentrations of glucose. This option allows you to expand the diagnostic capability of this test not only to confirm such a suspicion of diabetes, but now also for suspicion of tubulopathy. In the fourth study, quantitative method in chemistry liquid was evaluated for the determination of urinary bilirubin. Data obteined from urine samples of 194 dogs and 74 cats were compared with the dipstick results. The statistical analysis in samples of canine origin, however, has shown a statistically significant difference between the concentrations of bilirubin measured by the quantitative method and the three classes identified with dipstick. For the lack of positive samples by dipstick of cats urine, has not been possible to carry out a comparative statistical analysis. We observed that the dipstick is unreliable for the high rate of false negative and for the presence of overestimated and underestimated results in both species while the quantitative method in liquid chemistry has presented a good repeatability and linearity. The liquid chemistry method was being able to measure even low concentrations of urinary bilirubin. This is very important per una diagnosi precoce, especially in cats where the presence of bilirubinuria is always considered pathological. In the fifth study, we evaluated the kit for the quantitative determination of urinary nitrites in liquid chemistry in urine samples from 204 dogs and 72 cats in relation to different parameters and correlation with urinary tract infections. The high rate of false negative results when a dipstick is known for some time, but test strips remain the only instrument used in the routine diagnostic laboratory for the detection of nitrites in the urine. In addition, the ability to detect lower urinary concentrations of nitrite dipstick to those obtainable, opens new horizons. Currently, their importance in the clinical field, is tied exclusively to the diagnosis of urinary tract infections. It has been documented in numerous studies such as the presence of these ions may arise from the metabolism of nitric oxide and a small part also from the diet. Starting from the almost constant finding of nitrites in the urine, it is therefore sought to understand whether and how their concentrations might vary according to different characteristics of the urine. The ability to quantify the concentration of this analyte has led us to investigate the diagnostic utility of some parameters. In the sixth study, we applied the staining with Sudan III for the determination of lipiduria in 60 urine sediment (42 dogs and 12 cats). The evaluation of the urinary sediment after extemporaneous staining by optical microscopy showed that the lipid droplets consisted essentially of neutral fatty acids in both species. In addition, Sudan has been particularly useful to confirm the nature of the lipid inclusions present in cells and cylinders. At present it is difficult to assign them a certain pathological significance, however, we found these "oval fat bodies" only in sediments classified as pathological. It remains now the opportunity to apply different colors for easy identification elements potentially pathognomonic. In the seventh study, we wanted the qualitative assessment of haematuria. In human medicine, since the 70s, there have been numerous studies on the number of erythrocytes in the urine, red blood cell morphology and the relationship between MCV and MCV urinary blood in order to determine the origin of bleding. These parameters, especially the qualitative evaluation of haematuria (RBC dimorphism), are hardly taken into account in veterinary laboratory medicine despite considerable diagnostic potential. To this end, we evaluated the morphology of red blood cells in urine sediment by phase contrast microscopy. CONCLUSIONS - The urinalysis is probably the oldest question in the history of medicine, and still has a key role in the diagnostic laboratory. The technological history of chemical and physical examination of the urine, expressed in the second half of last century with the birth of the test strip (dipstick), which in clinical practice is still widely used. Over the years numerous studies have been conducted to assess the reliability, and although the results have highlighted the many limitations of this method is still used both in human and in veterinary medicine. Probably the lack of alternative economic and easy to use has allowed the maintenance of this custom. This, however, at the expense of the validity of the information obtained, which in many cases can be misleading in the diagnostic decision of a patient. In addition, the failure to apply standard procedures in the collection, storage, preparation and analysis of the sample made this examination poorly reproducible even within the same lab, increasing even more the unreliability of the test. This experimental work has highlighted the possibility of introducing alternative methods dipstick analysis. Quantitative methods in the chemical liquid, generally have the great advantage of spectrophotometric methods, also applicable to automated tools commonly used for biochemical analysis of serum calibrators and controls that allow you to keep check the analytical performance. The automation of the method eliminates operator dependent variables in the case of visual reading of the dipstick can greatly influence the result based on operator experience. It should be emphasized that even from an economic standpoint, the cost analysis by liquid chemical method is much less than the cost analysis by dipstick. The experimental work has focused on the quantitative determination of urinary following parameters: specific gravity, pH, glucose, bilirubin and nitrite. We want to remember as well that these parameters are of importance in veterinary laboratory medicine, that their determination in urine of dogs and cats dipstick has more limitations than those inherent in the method itself. In fact, the specific gravity determined by dipstick is almost unusable for the limited range of measurement, the pH appears inaccurate, especially for dog urine, glucose and bilirubin as well as being very sensitive in determining the analyte, has a high variable results over and under estimated without a trend identifiable nitrites however, have a high rate of false negatives in the dog and a high percentage of false positives in the cat. Analysis of the results we can say in general that all liquid chemical methods evaluated presented good repeatability and linearity. In particular, the determination of pH in chemical liquid showed a good correlation with the pH meter in both species, but it has some limits of detection in samples at pH less than about 5.5. We want to emphasize that the measurement range of the method, can detect pH between 4 and 10, are therefore also applicable to the evaluation of the urine of domestic large herbivores, whose physiological pH range between 7.5 and 8 in cattle and between 7.5 and 8.5 in the horse. The spectrophotometric method for the determination of glucose has allowed us to detect low concentrations of glucose. The statistical correlation of results with values obtained in dry chemical has been shown to lack. However we must emphasize that the paucity of positive samples may have affected the correlation test, in fact, the descriptive analysis of the results have not revealed large discrepancies between the two methods. The spectrophotometric method being more sensitive than dry chemical, allows an earlier diagnosis of glycosuria. Regarding the bilirubinuria, we can say that the method has found a good repeatability and linearity. In addition, a statistically significant difference was found between the concentrations of bilirubin (measured by spectrophotometry) in the three classes identified with disptick. We want to point out how many were found in the dipstick false negatives. Should be emphasized that the visual interpretation of this parameter can be affected by the presence of other chromogens. For this reason and for the applicability of the spectrophotometric method we need to re-evaluate the bilirubinuria, considering healthy animals and determining the physiological range bilirubinuria in dogs and cats. The method of detection of nitrite had a good repeatability and linearity. The statistical correlation between the chemical determinations of nitrite in dry and liquid chemical was not possible. Only 4 samples were positive dipstick, while the chemical liquid showed the presence of nitrite in almost all samples in our study. It has long been known to the inapplicability of the dipstick for the determination of nitrite in dogs and cats, due to the high rate of false negatives. For these reasons we preferred to consider the evaluation of nitrite in the liquid chemical in relation to the presence of bacteriuria by microscopic examination of sediment. The concentration of nitrite was significantly different for the presence of rods in the urine. the presence / absence of cocci. A significant variance was found only in the presence of "rod". This leaves open the possibility that these two species, as in humans, urinary nitrites do not come exclusively from bacterial metabolism, catabolism, but from which the endogenous nitric oxide. It would be interesting also to determine in a prospective study of healthy animals, the physiological concentration range of nitrite in urine. The determination of specific gravity of liquid in chemistry has proved unenforceable in dogs and cats for the limited range of the method. In addition, when compared with the refractometric evaluation showed poor correlation. It should be noted that the method, set up for human medicine, is capable of determining the specific gravity by weighing the chloride ions present in solution. It is possible that, for this reason, the method is not used to the dog and cat. We certainly need to find a component of urine that is more closely related to the specific weight in these two species. A study in human medicine, suggests the development of a method in chemical liquid for evaluating the specific weight, based on the determination of creatinine. Statistical analysis of the specific gravity evaluated refractometry and creatinine values we have seen that the variables are correlated, suggesting the development of a new method that considers the urinary creatinine as an index of specific gravity. The data analysis conducted between USG and then allowed UCr to develop a mathematical model, which can be a starting point for assessing the correlation between these two variables, but for now we can not say that the determination of specific gravity is possible by the determination of urinary creatinine. It is therefore considered desirable to continue the study for the development of chemical methods in the automation of the analysis of urine liquid times in dogs and cats, implementing the rose of the parameters currently available. Another key part of the analysis of urine from the urinary sediment is covered, but too often overlooked or misinterpreted. Even for a reliable measurement of sediment must be the application of standard procedures (ECLM, 2000) as it does for human medicine. The information obtained by the present microscopic evaluation may be extended by introducing a multi-modal reading of the sediment. In our experimental work we have seen how the application of colors impromptu allowed a qualitative assessment of lipiduria in dogs and cats. If it is considered pathological in the cat, the dog we saw how the presence of oval fat body and associated sediments "pathological." Further investigations are needed to examine these preliminary results. Furthermore, the application of phase contrast microscopy allowed us to evaluate the morphology of RBCs being haematuria,. Unfortunately, the lack of positive samples has limited the statistical evaluation of this study, but preliminary results comfort us in pursuing this path. The ability to determine the source of haematuria by morphological evaluation of RBCs would be unequivocally useful in the diagnostic field.

Carbon dots (CDs) are a novel carbon-based nanomaterial that draws a great attention in the last decade. The easy way of synthesis and the cheapest nature of the precursors used, stimulate a lot of scientist to propose new way of synthesis and applications. Notwithstanding the number of paper published, a systematic and reproducible way of synthesis is not yet been achieved, as well as a common definition of the resulting nanomaterials and their properties. In this thesis, the synthetic protocols and the characterization procedures used for the CDs will be deeply investigated in order to apply the nanomaterials in different applications. In particular, carbon dots synthesized from fully biocompatible precursors such as glucose, fructose and ascorbic acid, are characterized and employed for drug loading applications. The study reveals that the choice of the precursors is a crucial step because it affects the structural properties of the nanomaterials and the biological properties, revealing an unexpected toxicity for the fructose derived CDs, ascribable to its thermal degradation pathway. Furthermore, the drug loading capabilities were found to be correlated to the morphology of the nanoparticles, revealing the crucial role of the π-π interactions in achieving a loading up to 28 %wt for the glucose-based CDs. A second study was conduct on citric acid based CDs, in order to determine the best synthetic approach for photocatalytic applications. The nanoparticles synthesized both by hydrothermal and pyrolysis treatment, from sole citric acid and in combination with a nitrogen doping agent, were deeply characterized. The analysis reveals the difference in chemical, structural, and optical properties between the two synthetic methods. The photoreduction of methylviologen (MV) was used as model reaction to study the photocatalytic ability of the CDs, and the results reveal a relationship between the synthetic methods employed, the structural and optical properties of the CDs and their ability to act as a sensitizer. The amorphous nitrogen doped CDs reveals to be the best choice for this application with an initial rate conversion comparable to other reported in literature. The same CDs were also tested for the photocatalytic cleavage of C-O bonds in activated esters without the use of metals. The study reveals that CDs can successfully catalyze the reaction, with complete conversion and almost total selectivity. In addition, the CDs employed, shows different reactivity depending on the precursors and synthetic methods employed for their synthesis, with the same trend displayed for the MV photoreduction. This study highlights the ability of CDs to act as photosensitizer, without the addition of metals, in an organic reaction, opening a new scenario in the use of this nanomaterial in photocatalytic applications.

Negli ultimi anni, c’è stato un rapido sviluppo delle tecniche di crescita dei materiali nanostrutturati, e un forte impulso è stato dato dall’introduzione delle tecniche di crescita colloidale. Tali tecniche consentono di crescere un ampia gamma di materiali nanostrutturati, metalli e semiconduttori, con elevata cristallinità, dimensioni ridotte (< 5 nm) e con una distribuzione delle dimensioni molto stretta. Il solfuro di cadmio (CdS) nanostrutturato ha promettenti future applicazioni tecnologiche, come ad esempio nei dispositivi optoelettronici, celle solari ad alta efficienza e come tracciante fluorescente in biologia. Tuttavia, per poter sfruttare al meglio le proprietà fisiche a favore delle citate applicazioni è di fondamentale importanza una conoscenza approfondita delle proprietà fisiche. In questa tesi, sono state studiate le proprietà optovibrazionali e optoelettroniche dei quantum dots (QDs) di solfuro di cadmio cresciuti tramite un metodo di crescita colloidale. Tramite i metodi di crescita colloidale è possibile produrre QDs con dimensione ridotta e una distribuzione della dimensione molto stretta. La sintesi dei CdS-QDs consiste nella termolisi a circa 260 °C dello stearato di cadmio in presenza di solfuro di idrogeno in un solvente organico altobollente (1-ottadecene). La velocità della crescita e la dimensione finale dei QDs sono regolate dalla presenza di una molecola surfattante, l’ossido di triottilfosfina (TOPO). In particolare, QDs con una determinata dimensione e con una sua distribuzione molto stretta possono essere ottenuti regolando opportunamente la temperatura di crescita, la concentrazione dei precursori e principalmente la concentrazione del surfattante e del tempo di reazione (crescita arrestata). La morfologia, la dimensione (diametro) e la distribuzione dei diametri sono state determinate tramite TEM. Tramite spettroscopia di assorbimento, si ottengono informazioni sugli stati elettronici, inoltre, sfruttando la relazione esistente tra la band gap e il diametro, si può determinare il diametro medio di un campione di QDs. Le proprietà emissive dei QDs sono state studiate tramite spettroscopia di fotoluminescenza (PL) e dall’energia della banda di PL si può ottenere una stima del diametro medio dei QDs. Dalla larghezza di banda degli spettri di assorbimento e di PL si può ottenere anche una stima sulla distribuzione del diametro dei QDs. Un estesa parte del lavoro riguarda lo studio delle proprietà vibrazionali dei CdS-QDs, tramite spettroscopia Raman. Queste indagini sono state effettuate su campioni di CdS-QDs cresciuti appositamente con diversi diametri. Per eseguire misure micro-Raman, i campioni di CdS-QDs coordinati da molecole di TOPO che hanno una consistenza gelatinosa, sono stati trattati con acido tioglicolico (TGA). Questo trattamento è necessario per avere CdS-QDs in forma di polvere, la quale è più adatta per essere studiata tramite spettroscopia Raman. Per evitare effetti termici negli spettri o il danneggiamento del campione, le misure micro-Raman devono essere effettuate con potenze del laser molto basse. Negli spettri Raman di CdS-QDs si osserva uno spostamento del picco del fonone LO verso frequenze più basse , in particolare, tale spostamento è più marcato per i QDs più piccoli, mentre, al crescere del diametro, la frequenza si avvicina progressivamente a quella del bulk. Questa diminuzione di frequenza è causata dall’espansione del cristallo che avviene nei QDs, con il conseguente indebolimento dei legami per i quali diminuisce la frequenza di risonanza. Oltre a questo, il confinamento quantistico dei fononi è visibile come un allargamento asimmetrico della linea dei fononi, e come l’apparizione di un nuovo picco a circa 270 cm-1. Alcune pubblicazioni assegnano questo picco ai modi di superficie, mentre altri lo descivono come la conseguenza delle nuove regole di selezione dovute dalla bassa dimensionalità. Lo studio ha anche lo scopo di comparare le previsioni teoriche basate sia sul modello “dielectric continuum” che sui fononi di superficie con i risultati sperimentali. È stata trovata una relazione tra i valori di frequenza dei fononi predetti teoricamente e i risultati sperimentali, in particolare, le frequenze dei fononi di superficie sono in accordo con i risultati sperimentali. In conclusione, lo scopo di questo studio consiste nello sviluppo di un metodo per crescere CdS-QDs con le caratteristiche fisiche desiderate (diamtero voluto e distribuzione di diametro stretta) per poterne poi effettuare uno studio sistematico delle proprietà vibrazionali ed elettroniche.
In recent years, there has been a rapid development of the growth techniques of nanostructured materials, and a particular breakthrough was given by the introduction of colloidal growth techniques. These techniques allow to grow by affordable facilities, a wide range of nanostructured materials, metals and semiconductors, with high crystallinity, reduced size, narrow size distribution. Nanostructured cadmium sulfide (CdS) has promising future applications as in the realization of optoelectronic devices, high efficiency solar cells as well as fluorescent biological probe. However, in order to fully exploit the potential technological applications, the study of the physical properties of such materials is of crucial importance. In this thesis, the optoelectronic and optovibrational properties of cadmium sulfide quantum dots (QDs) grown by colloidal chemical method are studied. By the means of colloidal growth, it is possible to grow QDs with reduced size and narrow size distribution. The synthesis of CdS-QDs consists in the thermolysis (T=260 °C) of cadmium stearate in presence of hydrogen sulfide in a high temperature boiling point solvent (1-octadecene). The growth rate and final QDs size are regulated by the presence of the surfactating molecule trioctylphosphine oxide (TOPO). QDs with a determined size and a narrow size distribution can be obtained properly adjusting the growth parameters such as temperature, precursors concentrations, and principally the surfactant concentration and reaction time (arrested growth). The QDs morphology, their size and their size distribution is determined by TEM imaging. By absorption spectroscopy, information regarding the electronic states in QDs are obtained, and exploiting the relation existing between band gap and QD diameter, the mean diameter of the QDS is determined. The emissive properties of the QDs are probed by photoluminescence spectroscopy (PL). From the energy of PL band, an estimation of the QDs diameter can be obtained. Based on the width of absorbance and PL bands, the width of QDs size distributions can be estimated. A large part of the work is concerned with the study of vibrational properties of CdS-QDs by Raman spectroscopy. These investigations are carried out on the CdS-QDs samples purposely grown with different average sizes. In order to perform micro-Raman measurements, the gel-like TOPO-coated CdS-QDs are treated to replace the TOPO layer by thioglycolic acid (TGA). This treatment is necessary in order to have powder-like CdS-QDs being more suitable to a Raman scattering study. To avoid thermal effects or damage to the sample, the micro-Raman measurements must to be performed using very low laser powers (on the sample). In the Raman spectra of CdS-QDs, a decrease of the phonon frequency (red-shift) with respect to the bulk CdS frequency is observed. In particular, the red-shift is expected to be more pronounced for the smallest QDs, while at the increasing of QDs size, the phonon frequency will approach progressively to the bulk value. This red-shift is caused by the lattice expansion and by a subsequent weakening of the bonds which causes a reduction of the resonance frequency. Beyond the red-shift, the quantum confinement is visible also as an asymmetric broadening of the phonon line and by the apparition of a new peak a circa 270 cm-1. Some reports assign this peak to surface modes, while other reports describe this mode as a consequence of new selection rules arising from the reduced dimensionality. The study has also the aim to cross check the theoretical prediction based on the dielectric continuum model and on the surface modes with the experimental results. A relation between the theory and the experiment has been found, in particular, the predicted surface frequencies are in good agreement with the experiments. In conclusion, the goal of this thesis work is to develop a method to grow CdS-QDs with the desired physical characteristics (narrow size distribution) suitable for a systematic study of optical properties (vibrational and electronic).

Esta dissertação tem como tema o processo de reterritorialização das famílias rurais atingidas pela construção da barragem Barra Grande, situada no rio Pelotas, na divisa entre os estados do Rio Grande do Sul e Santa Catarina. A maioria da população rural atingida pertence a dois grupos socioculturais – caboclos e colonos (italianos) – e produzia essencialmente uma agricultura de subsistência. O objetivo é compreender a dinâmica de reorganização territorial, as mudanças nas unidades familiares produtivas e as consequências nos diferentes grupos socioculturais. A pesquisa foi conduzida sob uma perspectiva qualitativa, mediante utilização de entrevistas semiestruturadas, observação direta e documentação fotográfica, aliadas aos dados provenientes de fontes primárias e secundárias. Foram feitas entrevistas não estruturadas com base na perspectiva da história oral, procurando obter informações sobre o processo de territorialização-desterritorialização-reterritorialização (T-D-R), sobre as mudanças provocadas, a situação atual e a perspectiva das famílias. O universo empírico é constituído por agricultores deslocados compulsoriamente pela implantação da barragem que atualmente ocupam localidades e reassentamentos situados nos municípios de Anita Garibaldi/SC, Pinhal da Serra/RS e Esmeralda/RS. A territorialização diferente dos caboclos e dos colonos se expressa em configurações espaciais diferentes, mas a geografia da região ocupada, de relevo acidentado, faz com que haja aproximação do modo de produção agrícola, conhecido por roça (agricultura tradicional), com o uso de recursos disponíveis no ambiente. A construção da barragem, por sua vez, inicia o processo de desterritorialização tanto dos caboclos quanto dos colonos, tornando-os atingidos. Além do deslocamento compulsório dos agricultores, a barragem provoca impactos socioambientais, como a inundação das áreas agricultáveis e naturais, diminuindo o hábitat de espécies endêmicas da região. A reterritorialização é uma confluência do histórico de conflitos, das especificidades culturais e da organização ou não no movimento social. Há a perda de autonomia dos atingidos na reterritorialização, pois esta impõe mudanças nos usos da terra, com a substituição da roça, agricultura tradicional de subsistência, pela lavoura, agricultura moderna, altamente dependente de insumos externos (adubos, agrotóxicos e sementes); na relação e percepção do meio ambiente; no relevo, na paisagem e no microclima; e no uso de recursos naturais (água, lenha e frutas). A organização dos atingidos em um movimento social redefiniu a reterritorialização, principalmente na disputa em torno das modalidades de remanejamento populacional, sendo que, por parte do Movimento dos Atingidos por Barragens (MAB), a reivindicação era terra por terra (reassentamento), além do reconhecimento de todos os atingidos, disputando as modalidades indenizatórias propostas pela empresa (indenização em dinheiro e autorreassentamento por carta de crédito). A organização em um movimento social no meio rural através da mobilização e conscientização política acaba gerando outras formas de sociabilidade mais abertas com a sociedade regional, como associações produtivas, cooperativas, sindicatos rurais e partidos políticos.
The theme of this thesis is the process of reterritorialization of rural families affected by the construction of the Barra Grande dam, located in Pelotas River, on the border between the states of Rio Grande do Sul and Santa Catarina. Most of the affected rural population belongs to two socio-cultural groups – caboclos and (Italian) settlers – and produced an essentially subsistence farming. The goal is to understand the dynamics of territorial reorganization, changes in family production units and the consequences in different socio-cultural groups. The research was conducted under a qualitative perspective, by using semi-structured interviews, direct observation and photographic documentation, combined with data from primary and secondary sources. Non-structured interviews were conducted from the perspective of oral history, looking for information about the process of territorializationdeterritorialization- reterritorialization (T-D-R), the changes brought about, the current situation and perspective of families. The empirical universe is made up of farmers that were forcefully displaced by the construction of the dam and that currently occupy villages and settlements located in the municipalities of Anita Garibaldi/SC, Pinhal da Serra/RS and Esmeralda/RS. The different territorialization of caboclos and settlers has manifested itself in different spatial configurations, but the geography of the area occupied, with its broken terrain, causes an approximation to traditional farming, with the use of resources available from the environment. The construction of the dam in turn triggers the process of deterritorialization both of caboclos and settlers, affecting them. In addition to the compulsory displacement of farmers, the dam will cause social and environmental impacts, such as flooding of farmland and natural areas, thus reducing the habitat of endemic species in the region. A reterritorialization is a confluence of historical conflicts, cultural specificities and the presence or lack of organization in the social movement. There is loss of autonomy of those affected by the reterritorialization, as it demands changes in the uses of the land, by replacing the traditional subsistence farming with modern agriculture, which is largely dependent on external inputs (fertilizers, pesticides and seeds); in both the relationship with and the perception of the environment; in the topography, landscape and micro-climate; and in the use of natural resources (water, firewood and fruit). The organization of people affected in a social movement has redefined the reterritorialization, especially in the dispute over the methods of population relocation. The Movement of People Affected by Dams (MAB) was claiming land for land (resettlement), besides the acknowledgment of all those affected, competing for the modalities proposed by the company for damages (monetary indemnity and self-resettlement by means of letter of credit). The organization in a social movement in rural areas by mobilization and political awareness ends up generating other forms of sociability that are more open to the regional society, such as production associations, cooperatives, rural unions and political parties.

Since their first application, nanotechnologies have been capable to produce improvements and revolutionary results in many technological and industrial branches: medicine, transport, food security, environmental management and many others. Among these, nanotechnologies have proved their potential in medicine and in the treatment of polluted water. Nano-medicine made available increasing numbers of personalized and selective diagnostic methods and therapies. Nano-systems produced also more efficient purification processes for the treatment of polluted water. Thanks to the results obtained so far, these research fields are particularly significant and highly valued in the scientific world. With this in mind, the present thesis focused on assessing the application of Carbon Dots (CDs). CDs are nano-sized carbon particles, with dimensions between 1 and 10 nm. Since their discovery, they have proven their potential in countless fields. Here, CDs were evaluated for the treatment of polluted water and for their potential application in nano-medicine. In particular, their reduce toxicity makes them the ideal candidate for drug delivery purposes. In addition to this, their peculiar nature allows their employment as charge delocalizers and photosensitizers, implementing the properties of established water purification systems. Among these systems, TiO2 is, up to now, one of the most powerful material. When excited by the correct wavelength, TiO2 can react with the water absorbed on its surface and produce highly oxidative radicals. Radicals that are capable to decompose water pollutants. In this study, CDs have been synthesized from different precursors, and they have been characterized. The samples with the most promising properties have been selected and applied for the production of a TiO2-based high-performance photocatalytic material, capable of purifying polluted waters by exploiting solar light. This study pointed out how the formation of sub-products during the CDs synthesis may be the responsible for CDs cytotoxicity. The research demonstrated, moreover, that CDs cytotoxicity is not related to the toxicity of the starting reagents. The class of CDs, synthesized with citric acid and diethylenetriamine, displays the best properties for advanced TiO2 oxidation systems and the lowest cytotoxicity. Additionally, a TiO2-based photocatalytic system was also applied for water purification, exploiting the use of solar radiation. A high-performance photocatalytic material was produced by loading the CDs onto TiO2 particles. In order to preserve the CDs properties within the system, a hydrothermal process was developed and evaluated. In conclusion, we obtained a material which may be used to treat polluted water by exploiting the solar radiation. This material may represent the starting point for advanced water purification systems in developed and developing countries.

The purpose of this research is the problematisation in Adam Smith's moral philosophy of a classical question in the history of thought: what does it mean ‘to be human’ from a moral standpoint? In particular, I explored the relationship between the concepts of ‘human nature’ and ‘the self’ in order to reconstruct Smith’s theory of subjectivity, in the light of which I tried to show what it means ‘to be moral human beings’ in Smith’s philosophy. After providing a systematic reconstruction of Adam Smith’s conception of ‘human nature’ and ‘the self’ – exploring some aspects of Smith’s philosophy (e.g., sympathy, nature, human nature) and their empirical expressions (e.g., conduct) – I argued that, in Adam Smith’s work, the meaning of ‘moral human beings’ depends on the human being’s effort to live in harmony with oneself and the others. For Smith, this harmony can be reached by developing responsibly/morally one’s selfhood and satisfying «the most ardent desire» of human nature in society: the consciousness of being beloved or deservedly approved by oneself and the others. Finally, in Smith, ‘harmony with oneself and the others’ would be achieved in relation to a certain kind of awareness that can be possible when the human being tries to judge the conduct and tries to act according to the impartial spectator.

Now a days there is a world wide concern about the environment and this subject has affected all countries. However the commitment among the countries is not the same. The exploration of natural resources without rationality results in an environmental disaster, compromising the future of generations to come. Slashing the forest is a habit of the modern mankind that is the prejudicial not only to himself but also to other animals on earth. The settlers especially those from the Land Reform Program, organize themselves to preserve the environment after receiving the land. This study was focused in an investigation to understand the environmental knowledge of settlers from Darci Ribera s Project, located at Itaporanga County, to verify in loco, how they explain and organize themselves, in order to preserve the environment to reach a sustainable development. The work was based on theoretical concepts shown by Leff, Diegues and others thinkers, and was based on semi structured interviews, applications forms, and by observations of the day by day of settlers. The results obtained showed that settlers have a great amount of environmental knowledge, based in a potential of conservation and preservation, which has been passed from generation to generation, in how to use native vegetation not only as human food, but also as medicines, called by them jungle medicines . It was also pointed out, their productive capacity in order to provide surviving products, with a potential for the internal market, in order to promote a sustainable development. .
Na atualidade mundial a questão ambiental tem sido o assunto que tem afetado todos os países muito embora essa preocupação não se tenha por igual responsabilidade. A exploração dos recursos naturais provocando o desequilíbrio ambiental é a razão de ser das preocupações com o futuro da vida no Planeta e o desmatamento é a prática cultural do homem moderno mais nociva para a sua própria existência e dos demais seres bióticos. Os assentados, em particular aquele em Projetos de Assentamento da Reforma Agrária tem se organizado para conservar e ou preservar o meio ambiente após o recebimento da terra. O presente estudo teve como foco principal de investigação compreender os saberes ambientais dos assentados no Projeto de Assentamento Darci Ribeiro, localizado no município de Itaporanga D Ajuda/SE, na perspectiva de verificar in loco as formas explicativas da sua organização em relação com o meio ambiente e sua lógica de conservação e preservação de modo a promover o desenvolvimento sustentável. Para a concretização, o lume teórico revelado por Leff, Diegues e outros pensadores no assunto que direcionaram os rumos das investigações. Para tanto, foram realizadas por entrevistas semi-estruturadas, aplicação de questionários e da observação do cotidiano dos assentados. Os resultados alcançados revelaram que os assentados possuem um saber ambiental com base num potencial de preservação e conservação dos saberes adquiridos de geração a geração referente, em essência, ao uso da vegetação nativa tanto na alimentação humana como no uso e consumo de remédios ou remédios do mato , como dizem os assentados. Ressalta-se ainda sua capacidade produtiva como fonte abastecedora de produtos para a sua sobrevivência e em potencial para o mercado interno, promovendo o desenvolvimento sustentável.

In order to obtain safe and highly digestible foods destined to copanion animals the quality of the raw materials is essential. Food digestibility is a fundamental aspect in the pet foood industries, and particularly for the production of “complete foods”, that are opportunely formulated for the satisfaction of the entire animals nutritional needs. Dog food digestibility can be evalutated thorugh specific equations, through in vivo methods in which are employed animals, or in vitro methods, that simulate the passage of the food through the gastrointestinal tract, by using digestive enzymes. The majority of the pet foods sold nowadays are obtained by using the extrusion, a fast process that in few seconds can mix and heat raw materials (both dry and/or fresh) and turn into kibbles, that are already sanitized, and ready to be stabilizd through a drying phase. The extrusion process applies high temperatures (until 150°C) and pressure (40 atm), and in those conditions the digestibility of the material can be increased (starch gelatinisation, protein denaturation), but also the nutritional quality can be compormised by a decrease of available aminoacids (Maillard reaction) or available starch (starch retrogradation), fats oxydation and vitamin losses. A careful monitoring of the extrusion process is a key point in the assurance of the feed nutritional quality. In the present work the extrusion process for the dog foods production, the monitoring of the process parameters and the following evaluation of the digestibility, and the metabolic resoponses to the diets produced were studied. A further study regarded also the in vitro methods for the estimation of the food digestibility. This thesis is composed of an introductive bibliographic review and 3 experiments. The objective of the first contribution was the evaluation of the use of alpha amylases during the extrusion process, as possible mean for increasing the process procutivity. Two experiments were conducted, in each one the addiction of a different alpha amylase was tested. The productive parameters, along with the diets digestibility were evaluated. Resuld demonstrated an increase in extrusion productivity (until 40%) and a reduction of the energetic costs when the alpha amylase is added, without altering the food digestibility. The second contribution focused on the influence of the raw materials particle size and extrusion temperature on prodtive process and nutritional quality (digestibility, glycemic response and intestinal fermentations). To this purpose the same diet was ground using 4 different sieves mesh (0.5,0.8, 1.4 e 2 mm), and further extuded using two different extruder conformations,that allowed to reach two different temperatures (110 or 135°C). The 8 experimental diets obtained were then tested on 48 adult healthy dogs. Extrusion temperatures did not affect any parameter considered, while the raw material particle sze modified the productibity, that increased linearly to a particle size increase. If it is considered that in order to obtain finer particle sizes the energetic expense increases, the increase in productivity, along with a lower energetic demand for the grinding can be traduced in lower porductive costs. The in vivo tests demonstrated that to an increase in particle size corresponds a decrease in dry matter and protein digestibility, but also a higher microbial proliferation (significant linear increase of total aerobes, anaerobes and bifidobacteria). Moreover a lower fecal pH and ammonia, along with a higher SCFA production were registrated in animals fed diets with higher particle size. Those factors can contribute in keeping animals in healthy intestinal conditions. In the third contribute an in vitro 2 steps method (digestion with pepsin and pancreatin) was studied and some possible improvements were tested. A precipitation step with ethanol, before the recovery of the digested samples throug filtration, was introduced with the objective of increasing the soluble but not digested feed fraction, that otherwise wolud be lost during the filtration phase. Another objective of the study was to make the method easily reproducible in any simply equipped laboratory, to this purpose the in vitro procedure was carried out in a waterbath with manual or automatic agitation, instead of the equipment used in the original method, a thermostated camera with a magnetic stirrer . Samples used were previously tested for their in vivo digestibility. The in vitro estimation of the dog feed digestibility improved with ethanol precipitation, but a higher number of samples should be tested for the confirmation of an effective improvement. The use of a waterbath with manual agitation does not seem to interfere with the in vitro digestibility estimation; the reproducibility and repeatibility of the test conducted in waterbath is similar to that of the incubation equipment employed in the original method.
La qualità delle materie prime è un requisito fondamentale per ottenere alimenti per animali da compagnia sani e altamente digeribili. Soprattutto nel caso di alimenti “completi”, cioè studiati per soddisfare l’intero fabbisogno degli animali da compagnia, la digeribilità è un importante indice della qualità degli ingredienti ed un parametro di assoluto rilievo per l’industria del pet food. Essa può essere valutata attraverso l’uso di equazioni, metodiche in vivo che prevedono l’utilizzo di animali sperimentali e metodiche in vitro, che simulano il passaggio gastrointestinale degli alimenti mediante l’uso di enzimi digestivi. La quasi totalità degli alimenti destinati agli animali da compagnia è ottenuta attraverso l’estrusione, processo produttivo che consente in pochi secondi di passare dalla materia prima (farina o residui freschi dell’industria alimentare umana) al prodotto estruso, sanitizzato e pronto per essere stabilizzato tramite essiccazione. Il processo di estrusione prevede l’applicazione di alte temperature (fino a 150°C) e pressioni (fino a 40 atm), che possono sia migliorare la digeribilità (gelatinizzazione dell’amido, denaturazione proteica) degli alimenti, sia comprometterne la qualità nutrizionale (reazione di Maillard, retrogradazione dell’amido, ossidazione, perdita di vitamine). L’attento monitoraggio del processo di estrusione viene ad essere quindi un elemento basilare per poter garantire un prodotto nutrizionalmente corretto. Il presente lavoro di tesi ha avuto lo scopo di approfondire il processo di estrusione di alimenti per cani, monitorando il processo produttivo per verificarne l’impatto sulla digeribilità e le risposte metaboliche degli animali alle diete prodotte. Il primo contributo ha avuto come oggetto l’esecuzione di un processo estrusivo ai fini di valutare le ripercussioni dello stesso sulla materia prima aggiunta di due alfa amilasi differenti. Si è rilevato come l’aggiunta di tali enzimi al prodotto consente di raggiungere un aumento della produttività dell’estrusore ed una sensibile riduzione del costo energetico. Nel secondo contributo sperimentale è stata approfondita l’influenza delle dimensioni delle materie prime e della temperatura di estrusione sul processo produttivo e sulla qualità nutrizionale degli alimenti prodotti, espressa in termini di digeribilità e fermentazione intestinale. Nel presente esperimento è stata formulata una dieta di mantenimento per cani adulti, in seguito macinata utilizzando quattro setacci con granulometrie differenti. Le diete così ottenute sono state successivamente estruse utilizzando due diverse conformazioni dell’estrusore. Il piano sperimentale ha quindi previsto il test in vivo delle otto diete sperimentali su , 48 animali Le temperature di estrusione non hanno influito su nessun parametro. Al contrario, si è evidenziato una correlazione positiva tra granulometria e produttività. Considerando che l’energia impiegata per la macinazione aumenta di pari passo con la diminuzione della granulometria desiderata, si evince come si possa ottenere un’ulteriore diminuzione dei costi produttivi. L’esperimento in vivo ha dimostrato che ad un aumento delle dimensioni delle materie prime corrisponde una diminuzione della digeribilità di materia secca, proteine e grassi, ma anche una maggiore proliferazione microbica (batteri aerobi totali, anaerobi totali e bifido batteri), una diminuzione del pH e della concentrazione di ammoniaca fecale, ed una maggiore produzione di acidi grassi a catena corta, tutti fattori che migliorano la salute degli animali. Il lavoro descritto nel terzo contributo ha avuto come finalità l’implementazione di una metodica in vitro che prevede la simulazione del processo digestivo con enzimi quali pepsina e pancreatina. Attraverso l’introduzione di uno passaggio aggiuntivo di precipitazione con etanolo si è cercato di recuperare la componente solubile indigerita, che altrimenti verrebbe persa in fase di filtrazione. Si è inoltre cercato di rendere il metodo facilmente riproducibile anche con le strumentazioni più comuni; Infatti, in sostituzione della camera termostatata dotata di un agitatore magnetico utilizzata nel metodo originale, si è proceduto utilizzando un bagno termostatato in cui i campioni venivano agitati manualmente. Per tale lavoro, sono stati impiegati campioni di mangimi su cui erano già state eseguite precedenti analisi di digeribilità in vivo. L’introduzione della fase di precipitazione, ha favorito l’avvicinamento della stima di digeriblità in vitro al valore ottenuto in vivo. Inoltre, l’utilizzo del bagno termostatato come sistema di incubazione non sembra interferire con la stima della digeribilità in vitro e ha dimostrato una riproducibilità e ripetibilità del tutto paragonabili a quelle del metodo di incubazione originale. Grazie a questi incoraggianti primi risultati, sarebbe lecito pianificare in futuro l’applicazione dello stesso protocollo su una maggiore numerosità campionaria, al fine di affinare e irrobustire i risultati fin qui ottenuti.

The objective of our study is to evaluate the influence of colopexy on both clinical and surgical outcome of dogs treated with perineal herniorrhapy and, if possible, to establish its impact on rectal dilatation. Moreover our second aim is to compare the results of our surgical approach to those almost published in veterinary literature. Our preliminary hypothesis is that colopexy might improve prognosis of dogs treated with herniorrhapy and could at least reduce the severity of secondary rectal dilatation. According to our results colopexy, whether combined with herniorrhapy, seems to affect positively the surgical outcome of dogs with complicated perineal hernias. Patients treated with both colopexy and herniorrhaphy had relevant lower incidence of recurrence compared with dogs treated with the only herniorrhaphy. On the other hand it was not possible to quantify the impact of colopexy on rectal dilatation. To our opinion it was due to the fact that previously reported classification criteria, based over clinical reliefs, did not demonstrate enough accuracy in the evaluation of grade of rectal dilatation and no other diagnostic tool was available. Nevertheless we can disprove previous hypothesis(6) according which colopexy is effective in solving rectal dilatation since it was uncommon event in our study. Surprisingly a slight worse clinical outcome was assessed in dogs treated with colopexy since a significant percentage of dogs still had mild tenusmus in the postoperative period. We did not find any correlation between this aspect and data regarding the surgical outcome. However all the owners declared as good the overall quality of life of their dogs. It was not possible to perform a satisfying compare between our studies and other data previously published due to the huge heterogeneity of duration and methodology of follow up collection. Therefore we want to undermine the importance, in the near future, to develop and employ standardize criteria in both the preoperative classification and postoperative follow up evaluation in veterinary literature on perineal hernia treatment. Since the scarcity of the sample and the lack of strict inclusion criteria we have to consider our results as preliminary. Therefore additional studies with larger population are needed to confirm our data.

Abstract Study n. 1: Aujeszky Disease: serological and virological surveillance in Italy during 2012-2014 Although wild boar can act as a persistent Aujeszky’s disease (AD) reservoir, limited data are available on long-term epidemiology in free-ranging wild boar living in areas where industrial swine herds are limited. Hence, this study provides crucial information, which fills this knowledge gap, on the natural dynamics of AD infection. From 3260 sera sampled during eight hunting seasons, 162 (4.97%) were tested positive. Factors, including the animal’s age class, and the sampling year, had significant effects on the probability of the wild boar being seropositive, while wild boar mean abundance per area, yearly abundance and the total number of pig farms, as well as interactions among age, year and sex, were not significant. In particular, a positive trend of seroprevalence was observed over the years, with values ranging from 2.1 to 10.8%. This long-term surveillance showed an increase in seroprevalence with a higher probability of being seropositive in older individuals and the independence of wild boar seropositivity from the likelihood of contact with pigs in the area. Study n. 2: Genomic characterization of pseudorabies virus strains isolated from swine in Italy In this study, we undertook the genomic characterization of 44 pseudorabies virus (PRV) strains originated on pig farms isolated in Italy during 1984–2010. The characterization was based on partial sequencing of the UL44 (gC) and US8 (gE) genes. Thirty six porcine PRV strains, which were closely related to those isolated in Europe and America in the last 20 years belong to cluster B in both phylogenetic trees. Six porcine strains that do not belong to cluster B are related in both gE and gC phylogenetic trees to the ‘old’ porcine PRV strains isolated in the 1970s and 1980s. In the last two decades, the presence of these strains in domestic pig populations has been reduced drastically, whereas they are prevalent in wild boar. The two remaining strains have an interesting genomic profile, characterized by the gC gene being closely related to the old porcine PRV strains, and the gE gene being similar to that of recently isolated strains. These results provide interesting insight into the genomic characterization of PRV strains and reveal a clear differentiation between the strains isolated from wild boar and those originating from domestic pigs. Study n. 3: Detection and molecular analysis of pseudorabies virus strains isolated from dogs and wild boar in Italy Aujeszky’s disease (AD) is one of the most economically important diseases of farmed pigs. Wild boars can act as reservoirs and might represent a potential threat for domestic animals, including dogs. The aim of this study was to report the results of an AD survey based on the Pseudorabies virus (PRV) genome detection in samples of dogs clinically suspected of AD and of wild boars collected during four consecutive hunting seasons in the period 2010–2014. Genomic characterization was based on the partial gC sequence of the Italian strains and the comparison with those from domestic pigs and European PRV strains circulating in wild boars. The Italian PRV strains were mainly distributed into three different clusters and revealed two interesting findings. First, there was a clear distinction between the viral strains that were isolated from dogs used for hunting and subsequently traced back to wild boars and the strains that were isolated from working dogs and subsequently found to be closely related to domestic pigs. Second, the Italian epidemiological situation was found to be different from those of European countries in that the Italian situation was characterized by the presence of both the typical Italian clades 1 and 2 and supported by new patterns of aa deletions/insertions. Italian clade 1 included strains from hunting dogs and two Italian wild boars, and Italian clade 2 grouped with recent strains from dogs that were unable to hunt and domestic pigs that were related to one old reference strain (S66) and not included elsewhere. Molecular and phylogenetic analyses of PRV strains are therefore necessary to improve the understanding of the distribution of the PRV clusters and their evolution.

Aim - Veterinarians have frequently to deal with wounds to the skin, subcutis and underlying muscle. The aim was to explore the application of Hyaluronic Acid (HA) -containing dressing on open skin wounds in dogs. The progress of healing was assessed by wound area reduction and two scoring scales applied in human medicine. Materials and Methods - Ten client-owned dogs with 12 cutaneous open wounds healed by second intention were included. All wounds were treated using available in commerce HA-containing wound dressing from admission to complete re-epithelialization. At every clinical examination, wound area and scale scoring assessments were performed. Results - After debridement an increased wound size was obtained while an improvement was determined by both grading systems. The median numbers of return to clinic for bandage change was 5 times. The median time to complete wound healing was 34.5 days. The mean wound area at day 7,14,21 and 28 were respectively 90.4%, 47.7%, 22.4% and 14.8% of the original size (for linear measurement) and 95.5%, 54.4%, 23.10% and 14.8% of the original size (for software measurement). Regarding wound healing assessment tools, the agreement between two operators was considered high for both scales. Conclusions - HA-containing dressing may be a possible wound treatment for cutaneous open wounds in dogs. The assessment of wound quality using scale scoring system could be useful especially in the earlier healing period and to direct clinical decision-making process.