L'epatite E nei suidi: epidemiologia, diagnosi e filogenesi

Hepatitis E is an infectious viral disease with clinical and morphological features of acute hepatitis. The aetiological agent is the Hepatitis E virus (HEV). The disease represents an important Public Health problem in developing countries where is frequently epidemic and primarily transmitted by fecal-oral route. In the last few years, a certain number of sporadic cases have been also described in industrialized countries, Italy included. A swine HEV was first identified in 1997 and is now considered an ubiquitous virus. Human and swine strains from the same geographical region have shown to have a high level of nucleotidic omology and in experimental infections, the possibility of interspecific transmission of swine strains to humans and of human strains to non-human primates has been demonstrated. Furthermore, some seroepidemiological studies have demonstrated that people working in contact with swine have a higher risk to get infected than normal blood donors. Recently, cases of HEV hepatitis have been directly associated to the ingestion of uncooked tissues from pigs, wild boar or deer and today the disease is considered an emerging zoonosis. The aims of this thesis were: evaluate HEV prevalence in Italian swine herds (both in fattening and in breeding animals); investigate the possibility of finding HEV in livers used for human consumption; investigate if there is any correlation between HEV infection and the presence of macroscopical lesions; investigate HEV prevalence in a demographic managed wild boar population; phylogenetically analyse viral strains identified. During an internship period at Veterinary Laboratories Agency (Weybridge, UK), furthermore, swine samples at different stages of production and slurry lagoons have been analysed. Six swine herds located in North Italy have been sampled at different stage of production. The overall prevalence resulted 42%, and both breeding and fattening animals were positive for HEV infection. A longitudinal study has been conducted in a herd across all stages of production until the slaughtering age. Livers have been collected from the animals at the abattoir and 11.8% of them were positive for HEV infection. No correlations have been identified between HEV infection and macroscopical lesions in pigs affected by different pathological conditions. Of 86 wild boars tested 22 (25%) were positive for HEV. Of the swine tested in UK 21,5 % and 2 of the 9 slurry lagoons (22,2%) were positive for HEV infection. All the strains identified belonged to genotype 3 and showed high percentages of nucleotidic identity with humans and swine strains identified in Europe. The high prevalence detected in these studies confirms the widespread diffusion of HEV in swine populations in Italy and in UK. Phylogenetical analysis of identified strains, similar to those identified in autochthonous human hepatitis E cases of the same geographical area, confirm the hypothesis that pigs can be a font of zoonotical infection. The finding that a fraction of the livers inserted in the food chain are positive for HEV infection it’s of some concern for Public Health. The finding of a high HEV prevalence in all examined farms, together with the observation that infection may be sub-clinical and affect animals at slaughtering age, raise concern because of the possible risk of transmission of HEV to humans by either direct contact with infected pigs, indirect contact with environment and working instruments contaminated with pig feces, or ingestion of contaminated undercooked meat.

Eterogeneità delle proprietà fisico – chimiche della proteina prionica patologica e variabilità fenotipica ceppo specifica nella malattia di Creutzfeldt – Jakob

Transmissible spongiform encephalopathies (TSEs), or prion diseases, are neurodegenerative disorders that affect humans and mammals. Creutzfeldt-Jakob disease (CJD), the most common TSE in humans, can be sporadic (sCJD), genetic (gCJD), or acquired by infection. All TSEs are characterised by the accumulation of PrPSc, a misfolded form of the cellular protein PrPC. PrPSc is insoluble in detergents, partially resistant to proteolysis and shows a highly enriched β-sheet secondary structure. Six clinico-pathological phenotypes of sCJD have been characterized which correlate at the molecular level with two types (1 or 2) of PrPSc with distinctive physicochemical properties and the genotype at the polymorphic (methionine or valine) codon 129 of the prion protein gene. According to the protein-only hypothesis, which postulates that prions are composed exclusively of PrPSc, the strains of prions that are largely responsible for the wide spectrum of TSE phenotypes are enciphered in PrPSc conformation. In support to this view, studies mainly conducted in experimental scrapie, have shown that several prion strains can be identified based on distinguishing PrPSc biochemical properties. To further contribute to the understanding of the molecular basis of strains and to develop more sensitive strain typing assays in humans we have analyzed PrPSc biochemical properties in two experimental setting. In the first we compared the size of the core after protease digestion and the glycoform pattern of PrPSc before and after transmission of human prions to non human primates or bank voles, whereas in the second we analyzed the conformational stability of PrPSc associated with sCJD, vCJD or fCJD using guanidine hydrochloride (GdnHCl) as denaturant. Combining the results of the two studies, we were able to distinguish five human strains for at least one biochemical property. The present data extend our knowledge about the extent of strain variation and its relationship with PrPSc properties in human TSEs.

Integrative Musterbildung kranialer Strukturen

Form und Gestalt kraniofazialer Strukturen sind primär beeinflusst durch die inhärente Integration der unterschiedlichsten Funktionssysteme und externer selektiver Einflüsse. Die Variabilität der Schädel-Morphe ist ein Indikator für solche Einflussfaktoren und damit ein idealer Gegenstand für vergleichende Analysen morphogenetischer Formbildung. Zur Ermittlung morphologisch-adaptiver Trends und Muster wurden sowohl Hypothesen zur morphologischen Differenziertheit als auch zu Korrelationen zwischen modularen Schädelkompartimenten (fazial, neurokranial, basikranial) untersucht. Zusätzlich wurden aus Schichtröntgenaufnahmen (CT) virtuelle Modelle rekonstruiert, welche die Interpretation der statistischen Befunde unterstützen sollten. Zur Berechnung der Gestaltunterschiede wurden mittels eines mechanischen Gelenkarm-Messgerätes (MicroScribe-G2) max. 85 ektokraniale Messpunkte (Landmarks) bzw. dreidimensionale Koordinaten an ca. 520 Schädeln von fünf rezenten Gattungen der Überfamilie Hominoidea (Hylobates, Pongo, Gorilla, Pan und Homo) akquiriert. Aus dem Datensatz wurden geometrische Störfaktoren (Größe, Translation, Rotation) mathematisch eliminiert und die verbleibenden Residuale bzw. ‚Gestalt-Variablen‘ diversen multivariat-statistischen Verfahren unterzogen (Faktoren, Cluster-, Regressions- und Korrelationsanalysen sowie statistische Tests). Die angewandten Methoden erhalten die geometrische Information der Untersuchungsobjekte über alle Analyseschritte hinweg und werden unter der Bezeichnung „Geometric Morphometrics (GMM)“ als aktueller Ansatz der Morphometrie zusammengefasst. Für die unterschiedlichen Fragestellungen wurden spezifische Datensätze generiert. Es konnten diverse morphologische Trends und adaptive Muster mit Hilfe der Synthese statistischer Methoden und computer-basierter Rekonstruktionen aus den generierten Datensätzen ermittelt werden. Außerdem war es möglich, präzise zu rekonstruieren, welche kranialen Strukturen innerhalb der Stichprobe miteinander wechselwirken, einzigartige Variabilitäten repräsentieren oder eher homogen gestaltet sind. Die vorliegenden Befunde lassen erkennen, dass Fazial- und Neurokranium am stärksten miteinander korrelieren, während das Basikranium geringe Abhängigkeiten in Bezug auf Gesichts- oder Hirnschädelveränderungen zeigte. Das Basikranium erweist sich zudem bei den nicht-menschlichen Hominoidea und über alle Analysen hinweg als konservative und evolutiv-persistente Struktur mit dem geringsten Veränderungs-Potential. Juvenile Individuen zeigen eine hohe Affinität zueinander und zu Formen mit einem kleinem Gesichts- und großem Hirnschädel. Während das Kranium des rezenten Menschen primär von Enkephalisation und fazialer Retraktion (Orthognathisierung) dominiert ist und somit eine einzigartige Gestalt aufweist, zeigt sich der Kauapparat als maßgeblich formbildendes Kompartiment bei den nicht-menschlichen Formen. Die Verbindung von GMM mit den interaktiven Möglichkeiten computergenerierter Modelle erwies sich als valides Werkzeug zur Erfassung der aufgeworfenen Fragestellungen. Die Interpretation der Befunde ist durch massive Interkorrelationen der untersuchten Strukturen und der statistisch-mathematischen Prozeduren als hoch komplex zu kennzeichnen. Die Studie präsentiert einen innovativen Ansatz der modernen Morphometrie, welcher für zukünftige Untersuchungen im Bereich der kraniofazialen Gestaltanalyse ausgebaut werden könnte. Dabei verspricht die Verknüpfung mit ‚klassischen’ und modernen Zugängen (z. B. Molekularbiologie) gesteigerte Erkenntnismöglichkeiten für künftige morphometrische Fragestellungen.

Micro-simulation models at neighbourhood scale: the interplay of agents, buildings, activities and transport.

Urban systems consist of several interlinked sub-systems - social, economic, institutional and environmental – each representing a complex system of its own and affecting all the others at various structural and functional levels. An urban system is represented by a number of “human” agents, such as individuals and households, and “non-human” agents, such as buildings, establishments, transports, vehicles and infrastructures. These two categories of agents interact among them and simultaneously produce impact on the system they interact with. Try to understand the type of interactions, their spatial and temporal localisation to allow a very detailed simulation trough models, turn out to be a great effort and is the topic this research deals with. An analysis of urban system complexity is here presented and a state of the art review about the field of urban models is provided. Finally, six international models - MATSim, MobiSim, ANTONIN, TRANSIMS, UrbanSim, ILUTE - are illustrated and then compared.

Visual-somatosensory interactions in mental representations of the body and the face

The body is represented in the brain at levels that incorporate multisensory information. This thesis focused on interactions between vision and cutaneous sensations (i.e., touch and pain). Experiment 1 revealed that there are partially dissociable pathways for visual enhancement of touch (VET) depending upon whether one sees one’s own body or the body of another person. This indicates that VET, a seeming low-level effect on spatial tactile acuity, is actually sensitive to body identity. Experiments 2-4 explored the effect of viewing one’s own body on pain perception. They demonstrated that viewing the body biases pain intensity judgments irrespective of actual stimulus intensity, and, more importantly, reduces the discriminative capacities of the nociceptive pathway encoding noxious stimulus intensity. The latter effect only occurs if the pain-inducing event itself is not visible, suggesting that viewing the body alone and viewing a stimulus event on the body have distinct effects on cutaneous sensations. Experiment 5 replicated an enhancement of visual remapping of touch (VRT) when viewing fearful human faces being touched, and further demonstrated that VRT does not occur for observed touch on non-human faces, even fearful ones. This suggests that the facial expressions of non-human animals may not be simulated within the somatosensory system of the human observer in the same way that the facial expressions of other humans are. Finally, Experiment 6 examined the enfacement illusion, in which synchronous visuo-tactile inputs cause another’s face to be assimilated into the mental self-face representation. The strength of enfacement was not affected by the other’s facial expression, supporting an asymmetric relationship between processing of facial identity and facial expressions. Together, these studies indicate that multisensory representations of the body in the brain link low-level perceptual processes with the perception of emotional cues and body/face identity, and interact in complex ways depending upon contextual factors.

Analysen zur molekularen Charakterisierung von Proteinen des humanen Usher-Syndroms und Evaluation genbasierter Therapiestrategien

Analysen zur molekularen Charakterisierung von Proteinen des humanen Usher-Syndroms und Evaluation genbasierter Therapiestrategien rnDas humane Usher Syndrom (USH) ist die häufigste Form vererbter Taub-Blindheit. In der vorliegenden Dissertation wurde diese komplexe Erkrankung auf verschiedenen Ebenen analysiert: in Arbeiten zur Expression und Lokalisation von USH-Proteinen, der Analyse der USH-Proteinnetzwerke und deren Funktionen sowie darauf aufbauend die Entwicklung von Therapiestrategien für USH.rnIm Rahmen der Arbeit wurde die Expression und (sub)-zelluläre Lokalisation des USH1D-Genproduktes CDH23 in der Retina und Cochlea analysiert. CDH23-Isoformen werden in der Maus zeitlich und räumlich differentiell exprimiert. In den Retinae von Mäusen, nicht humanen Primaten und Menschen zeigten Analysen eine unterschiedliche Expression und Lokalisation des Zell-Zelladhäsionsmoleküls CDH23, was auf Funktions-unterschiede der einzelnen Isoformen in den analysierten Spezies hindeutet.rnAnalysen zur Aufklärung der USH-Proteinnetzwerke ergaben eine potentielle Interaktion des USH1G-Gerüstproteins SANS mit dem Golgi- und Centrosom-assoziierten Protein Myomegalin. Die direkte Interaktion der Proteine konnte durch unabhängige Experimente verifiziert werden. Beide Interaktionspartner sind in den Retinae verschiedener Spezies partiell ko-lokalisiert und partizipieren im periciliären USH-Proteinnetzwerk. Die Assoziation von SANS und Myomegalin mit dem Mikrotubuli-Cytoskelett weist auf eine Funktion des Proteinkomplexes in gerichteten Transportprozessen innerhalb der Photorezeptoren hin und bekräftigt die Hypothese einer Rolle von SANS und assoziierten Netzwerken mit Transportprozessen.rnDas hier gewonnene erweiterte Verständnis der molekularen Grundlagen sowie die Aufklärung der zellulären Funktion der Proteinnetzwerke ermöglichen die Entwicklung therapeutischer Strategien für USH. Ein Fokus der vorliegenden Arbeit lag auf der Entwicklung genbasierter Therapiestrategien und deren Evaluation, wobei der Schwerpunkt auf der Therapiestrategie der Genreparatur lag. Die mit Hilfe von Zinkfinger-Nukleasen (ZFN) induzierte Homologe Rekombination für die Genkorrektur wurde exemplarisch an der 91C>T/p.R31X-Mutation im USH1C-Gen gezeigt. Effiziente ZFN wurden identifiziert, generiert und erfolgreich im Zellkulturmodellsystem eingesetzt. Die Analysen demonstrierten eine Reparatur der Mutation durch Homologe Rekombination auf genomischer Ebene und die Expression des wiederhergestellten Proteins. Durch die Genkorrektur im endogenen Lokus sind Größe des Gens, Isoformen oder die Art der Mutation keine limitierenden Faktoren für die Therapie. Die in der vorliegenden Arbeit durchgeführten Experimente unterstreichen das enorme Potential ZFN-basierter Therapiestrategien hin zu personalisierten Therapieformen nicht nur für USH sondern auch für andere erbliche Erkrankungen, deren genetische Grundlagen bekannt sind.rn

Why do we yawn? The importance of evidence for specific yawn-induced effects

Gallup (this issue) believes that our recent review on the function of yawning (Guggisberg et al., 2010) is unbalanced and that it ignores evidence for his thermoregulation hypothesis. Here we address these criticisms and show them to be untenable. While we never claimed that the social hypothesis of yawning has "definite experimental support", we emphasize the importance of experimental evidence for specific effects of yawns when considering why we yawn. The only specific effect of yawning that could be demonstrated so far is its contagiousness in humans, some non-human primates, and possibly dogs, whereas all studies investigating physiological consequences of yawns were unable to observe specific yawn-induced effects in the individual of any species. The argument that from an evolutionary perspective, yawns must have a "primitive" physiological function arises from imprecise reasoning.

Capuchin Monkeys Exercise Self-control by Choosing Token Exchange Over an Immediate Reward

Self-control is a prerequisite for complex cognitive processes such as cooperation and planning. As such, comparative studies of self-control may help elucidate the evolutionary origin of these capacities. A variety of methods have been developed to test for self-control in non-human primates that include some variation of foregoing an immediate reward in order to gain a more favorable reward. We used a token exchange paradigm to test for self-control in capuchin monkeys (Cebus apella). Animals were trained that particular tokens could be exchanged for food items worth different values. To test for self-control, a monkey was provided with a token that was associated with a lower-value food. When the monkey exchanged the token, the experimenter provided the monkey with a choice between the lower-value food item associated with the token or another token that was associated with a higher-value food. If the monkey chose the token, they could then exchange it for the higher-value food. Of seven monkeys trained to exchange tokens, five demonstrated that they attributed value to the tokens by differentially selecting tokens for higher-value foods over tokens for lower-value foods. When provided with a choice between a food item or a token for a higher-value food, two monkeys selected the token significantly more than expected by chance. The ability of capuchin monkeys to forego an immediate food reward and select a token that could then be traded for a more preferred food demonstrated some degree of self-control. Thus, results suggest a token exchange paradigm could be a successful technique for assessing self-control in this New World species.

The Savannah hypotheses: origin, reception and impact on paleoanthropology

The reconstruction of the human past is a complex task characterized by a high level of interdisciplinarity. How do scientists from different fields reach consensus on crucial aspects of paleoanthropological research? The present paper explores this question through an historical analysis of the origin, development, and reception of the savannah hypotheses (SHs). We show that this model neglected to investigate crucial biological aspects which appeared to be irrelevant in scenarios depicting early hominins evolving in arid or semi-arid open plains. For instance, the exploitation of aquatic food resources and other aspects of hominin interaction with water were largely ignored in classical paleoanthropology. These topics became central to alternative ideas on human evolution known as aquatic hypotheses. Since the aquatic model is commonly regarded as highly controversial, its rejection led to a stigmatization of the whole spectrum of topics around water use in non-human hominoids and hominins. We argue that this bias represents a serious hindrance to a comprehensive reconstruction of the human past. Progress in this field depends on clear differentiation between hypotheses proposed to contextualize early hominin evolution in specific environmental settings and research topics which demand the investigation of all relevant facets of early hominins' interaction with complex landscapes.

Nature, Poetry, and Liminality in the Poetry of Robert Frost

Explores how Frost examines and configures the divide between life’s imperfections and its rewards. I am particularly interested in how Frost positions both the non-human natural world and poetry itself as intermediary (or liminal) realms that might help us live simultaneously in the worlds of reality and the imagination, or truth and beauty, or heaven and earth.

The contexts of scratching behavior and postconflict behavior in squirrel monkeys (Saimiri sciureus)

Self-directed behavior (SDB), such as scratching, is a reliable indicator of emotional arousal in non-human primates. In contrast, affiliative behavior, such as social grooming, has been shown to have a calming effect in primates and reduce arousal. In order to test whether the expression of SDB was related to arousal, the scratching behavior of eight captive squirrel monkeys (Saimiri sciureus) was compared across four social contexts (huddling, proximity to others, solitary and post-conflict). In addition,rates of scratching were examined before and after affiliative behavior during the postconflict context. I tested for this effect by using the post-conflict/matched control(PC/MC) method in which post-conflict (PC) behavior of an animal is compared to thebehavior of the same animal in a baseline, nonaggressive situation or a matched control(MC). Context and associated scratching data were obtained from a total of 98 hours of focal sample data. Scratching was significantly lower while animals were huddling thanthe other two contexts. Scratching rates while solitary were significantly higher than those occurring while animals were in proximity. Scratching was also higher in PC than MC. Following conflict, animals were significantly more likely to make contact withthird parties not involved in aggression. Most of these (79%) were a third party approaching a combatant. Further, scratching rates decreased following post-conflict third party contacts and the decrease was not due to a general decrease in scratching thatmight have been occurring after the aggressive interaction. Huddling behavior appears to reduce arousal in squirrel monkeys and may act as a tension-reduction mechanism. The elevated scratching in the solitary context may suggest that squirrel monkeys may be engaged in activities while solitary, such as vigilant behavior that may increase arousal. The third party post conflict affiliative contacts observed were the first such interactions observed in squirrel monkeys. The fact that these third contacts reduced scratching ratesin the combatants indicates that 'consolation' may have been demonstrated in this species. The overall pattern of results suggested that scratching was reliable behavioral indicator of anxiety in squirrel monkeys. These results indicate that overt behavior can be used to assess emotional states in this and other species, acting as a mediator to understanding how emotions regulate social behavior.

Brown Capuchins (Cebus apella) Exhibit Self-Control on a Delay of Gratification Food Exchange Task When Food Options Differ Qualitatively

Self-control allows an individual to obtain a more preferred outcome by forgoing an immediate interest. Self-control is an advanced cognitive process because it involves the ability to weigh the costs and benefits of impulsive versus restrained behavior, determine the consequences of such behavior, and make decisions based on the most advantageous course of action. Self-control has been thoroughly explored in Old World primates, but less so in New World monkeys. There are many ways to test self-control abilities in non-human primates, including exchange tasks in which an animal must forgo an immediate, less preferred reward to receive a delayed, more preferred reward. I examined the self-control abilities of six capuchin monkeys using a task in which a monkey was given a less preferred food and was required to wait a delay interval to trade the fully intact less preferred food for a qualitatively higher, more preferred food. Partially eaten pieces of the less preferred food were not rewarded, and delay intervals increased on an individual basis based on performance. All six monkeys were successful in inhibiting impulsivity and trading a less preferred food for a more preferred food at the end of a delay interval. The maximum duration each subject postponed gratification instead of responding impulsively was considered their delay tolerance. This study was the first to show that monkeys could inhibit impulsivity in a delay of gratification food exchange task in which the immediate and delayed food options differed qualitatively and a partially eaten less preferred food was not rewarded with the more preferred food at the end of a delay interval. These results show that New World monkeys possess advanced cognitive abilities similar to those of Old World primates.

The tragedy of the commons in evolutionary biology

Garrett Hardin's tragedy of the commons is an analogy that shows how individuals driven by self-interest can end up destroying the resource upon which they all depend. The proposed solutions for humans rely on highly advanced skills such as negotiation, which raises the question of how non-human organisms manage to resolve similar tragedies. In recent years, this question has promoted evolutionary biologists to apply the tragedy of the commons to a wide range of biological systems. Here, we provide tools to categorize different types of tragedy and review different mechanisms, including kinship, policing and diminishing returns that can resolve conflicts that could otherwise end in tragedy. A central open question, however, is how often biological systems are able to resolve these scenarios rather than drive themselves extinct through individual-level selection favouring self-interested behaviours.

Sweet Puppies and Cute Babies: Perceptual Adaptation to Babyfacedness Transfers across Species

Infant faces are very salient stimuli. The Kindchenschema describes specific features that characterize a cute infant face. In this study we used a visual adaptation paradigm to investigate the universality of the perceptual properties of the Kindchenschema. In Experiment 1, twenty-four participants adapted to cute and less cute human infant faces and in Experiment 2, twenty-four new participants adapted to cute and less cute faces of puppy dogs. In both experiments the task was to assess the cuteness of subsequently presented human infant faces. The results revealed cuteness after-effects for human infant faces in both adaptation conditions, suggesting a common mechanism coding cuteness in human and non-human faces. This study provides experimental evidence for the universality of the well-described concept of the Kindchenschema.

Change Detection Memory in Rhesus Monkeys and Humans

Visual short-term memory (VSTM) is the storage of visual information over a brief time period (usually a few seconds or less). Over the past decade, the most popular task for studying VSTM in humans has been the change detection task. In this task, subjects must remember several visual items per trial in order to identify a change following a brief delay interval. Results from change detection tasks have shown that VSTM is limited; humans are only able to accurately hold a few visual items in mind over a brief delay. However, there has been much debate in regard to the structure or cause of these limitations. The two most popular conceptualizations of VSTM limitations in recent years have been the fixed-capacity model and the continuous-resource model. The fixed-capacity model proposes a discrete limit on the total number of visual items that can be stored in VSTM. The continuous-resource model proposes a continuous-resource that can be allocated among many visual items in VSTM, with noise in item memory increasing as the number of items to be remembered increases. While VSTM is far from being completely understood in humans, even less is known about VSTM in non-human animals, including the rhesus monkey (Macaca mulatta). Given that rhesus monkeys are the premier medical model for humans, it is important to understand their VSTM if they are to contribute to understanding human memory. The primary goals of this study were to train and test rhesus monkeys and humans in change detection in order to directly compare VSTM between the two species and explore the possibility that direct species comparison might shed light on the fixed-capacity vs. continuous-resource models of VSTM. The comparative results suggest qualitatively similar VSTM for the two species through converging evidence supporting the continuous-resource model and thereby establish rhesus monkeys as a good system for exploring neurophysiological correlates of VSTM.