Retirement Systems, Demography, Happiness and Welfare Redistribution

This research investigates whether an equity improvement within retirement-systems domain may positively influence demography, people’s happiness and their financial conditions. In particular, a fertility-boosting policy has been tested, acting on the contributory rate. This project has been carried out by using software simulation and with specific Agent-based Computational Economics (ACE) methodology. Two virtual worlds have been created, in order to try to reproduce Italian society. In the first model, (W1), vertical equity has been improved, while in the second one, (W2), it is has not. Five further variants of these two worlds have been produced by altering some parameters, in order to test our hypothesis through several simulations. The research outcomes prove that an equity improvement can positively influence demographic trends, can increase the level of happiness in the society, and can grant a more homogeneous welfare redistribution.Retirement systems, demography, happiness, wealth distribution, equity, software simulation, Agent Based Computational Economics

Investigating reward-based motor performance in volatile environments using computational modelling and electroencephalography

Motor improvements have been linked to reward magnitude in deterministic contexts. Nevertheless, it remains unclear whether individual inferences about reward probability dynamically influence motor vigour. Moreover, how factors such as age, Parkinson’s disease or anxiety affect the modulation of motor vigour by predictions of reward probability remains unexplored. This thesis, across four experiments, investigates how inferences about the volatile action-reward contingencies modulate motor performance on a trial-by-trial basis. We employed a reward-based motor decision-making task and modelled the behavioural data using the Hierarchical Gaussian Filter (HGF). In the final two studies, we also recorded the brain electrical activity through electroencephalography and used convolution models for oscillatory responses to delve into the neural underpinnings of motor decisions. The results revealed that stronger predictions about action-reward probabilities led to faster performance tempo on a trial-by-trial basis in healthy participants. This effect was preserved in older adults and medicated Parkinson’s disease patients. Furthermore, the invigoration of motor responses extended to explicit beliefs (confidence) about reward tendencies. Trait anxiety did not modulate the association between predictions and motor performance but affected practice effects over time. Analyses of the time-frequency representation of HGF computational quantities describing decision making unveiled increased alpha/beta correlates of different types of uncertainty among high trait anxiety individuals. Finally, we found that state anxiety dampened the invigoration effect previously discussed. This manifested as longer reaction time for actions that were highly anticipated to yield rewards. Moreover, state anxiety led to reduced theta oscillatory responses during processing win/lose outcomes. In conclusion, this thesis integrates computational modelling, Bayesian statistics, and electrophysiological approaches to explore motor decision-making behaviour under volatility. It provides novel evidence for an invigoration of motor performance by predictions about the action-reward contingency and sheds light on the modulation of this effect by age, Parkinson’s disease, trait and state anxiety

TESTUDINID HERPESVIRUS 3: DETECTION AND MOLECULAR CHARACTERIZATION OF STRAINS IN ITALIAN TESTUDO SPP.

Gli Herpesvirales sono un gruppo di virus a DNA ontogeneticamente antico, in grado di infettare diverse specie animali inclusi i rettili. Tra le diverse famiglie di herpesvirus gli Alphaherpesvirus sono la sottofamiglia di Herpesvirales maggiormente coinvolta nelle infezioni di rettili e sono re-sponsabili di lesioni necrotizzanti di gravit\ue0 variabile a carico di diversi organi; spesso le lesioni portano a morte il soggetto sia a causa dei danni indotti direttamente dal virus che per l'insorgenza di infezioni batteriche secondarie. Tra tutte le specie di rettili le tartarughe, le lucer-tole e i serpenti sono gli animali domestici pi\uf9 comuni. I proprietari di tartarughe sono, tra le tipologie di proprietari di rettili, quelli pi\uf9 proni ad acquistare animali prelevati in natura e ad abbandonare specie alloctone sul territorio. Inoltre, sono presenti numerosi allevamenti amatoriali di queste specie che non rispettano le buone norme di allevamento e introducono animali senza effettuare regolare quarantena. La somma di questi comportamenti ha avuto molteplici conse-guenze, tra le quali la diffusione di malattie esotiche e non esotiche nelle popolazioni autoctone di cheloni. Tra tutti i virus che possono causare infezioni nelle tartarughe, gli Herpesvirus sono i maggior-mente rappresentati, con 5 gruppi in grado di infettare testuggini terrestri e acquatiche. Inoltre, sulla base di quanto riportato in letteratura, gli herpesvirus sono una delle principali cause di morte tra i cheloni. Sebbene questi agenti eziologici abbiano un ruolo di rilievo sia da un punto di vista della bioconservazione che economico, non sono presenti informazioni sulla presenza del virus in Italia e, in linea generale, pochissime informazioni sono disponibili sulle caratteristiche genetiche e sui meccanismi di interazione ospite-patogeno. Questo lavoro ha lo scopo di identificare la presenza di Testudinid herpesvirus (TeHVs) in Italia, caratterizzare il genoma del virus ed iniziare ad investigare i meccanismi di interazione ospi-te-patogeno tra TeHV3 e Testudo graeca. Per valutare la presenza dei TeHVs in Italia, abbiamo effettuato uno studio prospettico effet-tuando test ELISA e PCR su tutte le tartarughe presenti nell'Oasi WWF di Vanzago ed abbiamo fatto, inoltre, uno studio retrospettivo effettuando la PCR sui campioni d'archivio dell'Istituto di Anatomia Patologica Veterinaria dell'Universit\ue0 di Milano. Esame necroscopico ed istopatologi-co completo sono stati effettuati su tutti i soggetti deceduti spontaneamente provenienti dall'Oasi di Vanzago. Inoltre, da uno dei soggetti provenienti dall'Oasi \ue8 stato possibile effettuare l'isolamento virale. Utilizzando ceppi di TeHV3 provenienti da Italia, USA e Svizzera, \ue8 stato possibile sequenziare il genoma di TeHV3. La valutazione dell'interazione ospite-patogeno \ue8 stata fatta utilizzando una libreria fagica screenata con siero iperimmune di T. graeca, ottenuto da uno studio di trasmissione precedente. Il nostro studio ha dimostrato che tutti i campioni positivi, sia prospettici che retrospettivi, erano positivi per TeHV3, tranne un caso retrospettivo positivo per TeHV1. . Il genoma di TeHV3 \ue8 risultato essere lungo, circa, 150.080 nucleotidi, con una configurazione genomica di tipo D ed \ue8 risultato altamente colineare con il genoma dell'Human herpesvirus 1. Da un punto di vista immunologico, abbiamo individuato tre potenziali proteine responsabili della risposta immunitaria dell'ospite, TE-17, UL-15 e la gB. Sebbene si fosse inizialmente ritenuto che la gB fosse la principale proteina responsabile della risposta immunitaria, valutazioni successive hanno dimostrato che l'orientamento della sequenza che codifica per la gB nel fagemide fosse antisenso e che, quindi, non potesse essere trascritta. Per valutare il possibile ruolo antigenico di gB, il fagemide originale \ue8 stato modificato in modo tale da contenere un solo frammento codificante. Le tecniche utilizzate per modificare il fagemide sono state: restrizione enzimatica, clonaggio con PCR e direct site mutagenesis con PCR. Sebbene siano stati fatti numerosi tentativi e utilizzate metodologie differenti, a tutt'oggi, non \ue8 stato possibile ottenere colonie batteriche contenti il fagemide modificato e, quindi, non \ue8 stato possibile confermare l'ipotesi originale.Herpesvirales is an ancient group of DNA viruses that infect different animal species, including reptiles. The Alphaherpesvirus is the major subfamily of Herpesvirales involved in reptile infections, and responsible for the onset of variably severe necrotizing lesions, affecting different body parts, and frequently associated with the animal's death. The death of the animal can be caused directly by the effect of the virus, or more frequently by the onset of secondary bacterial infections. Today, the reptile species most commonly kept as household pets are tortoises, lizards, and snakes. Compared to the owners of the other two groups of reptiles, at least in Europe, pet tortoise owners frequently buy animals that are collected from the wild (especially exotic species) and are also prone to abandon these animals into the wild. Moreover, good breeding practices are frequently not applied to reptile breeding centers, which commonly introduce new animals in their enclosure without testing and/or quarantining the animals. These habits have different consequences, including spreading both exotic and non-exotic diseases on Italian soil. Among all viruses infecting chelonians, herpesviruses are the group with the highest number of members, with more than five groups affecting turtles and tortoises. International literature re-ports herpesviruses as one of the most relevant causes of death in chelonians. Despite the relevance of the disease from both a conservational and economical point of view, no information is available on the presence of these viruses in Italy, and very little is known about the genetic characteristics and the host-pathogen interaction mechanisms. This work aims to identify the presence of Testudinid herpesviruses (TeHVs) in Italy, characterize the genome of the virus, and start to understand the host-pathogen interaction mechanisms between TeHV3 and Testudo graeca. To evaluate the presence of TeHVs in Italy, we carried out a prospective study, performing the ELISA test on live animals in the WWF Vanzago's Oasis, combined with PCR on both prospective and retrospective samples collected from the archive of the anatomical pathology section of the University of Milan. Complete necropsy and histological evaluation were performed on all prospective samples after their natural death. Furthermore, viral isolation was successfully carried out on one of the Vanzago cases. Using both TeHV3 strains collected in Italy, USA, and Switzerland, complete genome sequencing was performed. Evaluation of T. graeca immune response against TeHV3 was evaluated by creating a bacteriophage expression library screened with hyperimmune tortoise sera, obtained from a previous transmission study. Our study demonstrates that from both the prospective and retrospective samples, all that were positive were TeHV3, but one retrospective sample was TeHV1 positive. TeHV3 genome sequencing demonstrated that the viral genome is at least 150,080 nucleotides long, arranged in a D-type configuration, and extensively co-linear with the human herpesvirus 1 genome. From an immunological point of view, we were able to identify three relevant TeHV3 candidate antigenic proteins, TE-17, UL-15, and gB. Although we initially supposed the gB was the most relevant antigenic protein in T. graeca immune response against TeHV3, further investigation showed that the gB sequence in the evaluated phagemid was antisense compared to the origin of replication, and could not be transcribed. To assess the possible immunological relevance of TeHV3 gB protein in boosting host immune response, we performed enzymatic restriction, PCR cloning and PCR direct site mutagenesis on the original phagemid, to obtain new structures containing only one of the three possible immunogenic proteins previously identified. Despite the numerous attempts and techniques used, we are currently not able to obtain viable bacteria to confirm our hypothesis

Characterization of the immune response against Testudinid herpesvirus 3: new insight

Testudinid herpesvirus 3 (TeHV3) is one of the most lethal viral agents in tortoises worldwide. Although TeHV3 have been extensively studied, only little information is available about host-pathogen interaction. TeHV3 infections in different species of the genus Testudo correlate with various lesions profiles, disease severity and clinical outcome, suggesting the existence of a complex host-pathogen interaction. This might reflect a possible viral-host coevolution (Origgi, 2012).To study the host-pathogen interaction, we previously screened 5.000 clones from a bacteriophage library obtained from the TeHV3 genomic DNA using Testudo graeca seropositive sera. Of the six detected positive clones, only one was confirmed by F.A.C.S. Selected clone was determined to be a concatamer of different TeHV3 genomic fragments including the partial sequence of TE17, UL15, Major capsid protein (MCP), and Glycoprotein B (gB) genes. After complete sequencing of the selected clone, the MCP and the gB were antisenses compared to the phagemid promoter.In order to assess which of the gene fragments among TE17 and UL15 was encoding for the antigenic determinant that was recognized by the anti-TeHV3 tortoise sera, distinct approaches were followed.TE-17 and UL15 fragments were knock out from the original phagemid using the following approaches: a) directed-site mutagenesis, b) molecular cloning, and c) restriction enzymes cloning. All the modified constructs were cloned in two different E. coli cloning vectors (D5α and XL 1-Blue).Transformation of competent cells with the constructs described above did not yield any viable bacteria.Among the different aspects might have influenced transformation success rate, construct size was probably the most relevant (about 9Kb). Furthermore, we could not entirely exclude that genomic DNA editing might have induced mutations in the construct sequence causing toxic effects on the host bacterial cell. Cloning of TE-17 and UL15 gene fragments into different prokaryotic expression vectors is currently under way

CyPLOS: a new family of synthetic ionophores

The ion transport properties of a new family of synthetic ionophores based on cyclic phosphate-linked oligosaccharide (CyPLOS) macrocycles are described

Characterization of the immune response against Testudinid herpesvirus 3.

Numerous infectious diseases have been documented in reptiles, however minimal information isavailable concerning their immunological response. One of the most diffuse and lethal reptile pathogenis Testudinid herpesvirus 3 (TeHV3), a Alphaherpesvirinae. All species of tortoises (Testudinide) areconsidered susceptible to TeHV3, however the virus is over represented in the genus Testudo, whichincludes, among others, T. graeca, T. hermanni, T. marginata, and T. horsfieldii, that are popular pets inEurope. Incidence of TeHV3-associated disease is highest right after hibernation (Origgi, 2012).The aim of this work is to partially characterize the immunological response of T. graeca against TeHV3.A bacteriophage library composed of about 5.000 clones containing genomic DNA fragments of TeHV3was produced. Bacteriophages were amplified in a specific strain of E. coli and were screened withTeHV3-seropositive sera from T. graeca. Phagemids were excised from the positive bacteriophages,sequenced, and compare with the TeHV3 genome to identify the encoding genes. Six differentstructural and non-structural proteins have identified as immune relevant. Vero cells where transfectedwith phagemids of the positive clones, to confirm previous results. TeHV3’s proteins expression wasassessed by F.A.C.S using T. graeca seropositive sera. Of all the six selected clones, only that expressingthe partial sequence of the glycoprotein B (gB) showed a positive signal in the F.A.C.S. analysis. Thisresult is consistent with the well-known immunogenicity of gB of other herpesviruses including thoseinfecting humans and with the highly conserved role that gB plays in host-pathogen interaction acrossspecies and evolution (Beals et al., 2016)

Synthesis and characterization of trans-di-(4-pyridyl)porphyrin dimers

Preparation and characterization of a small library of symmetric trans-di(4-pyridyl) porphyrin dimers, obtained by either Glaser\u2013Hay or Sonogashira coupling reactions from appropriately prepared trans-di-4-pyridylporphyrin precursors, is presented. The porphyrin dimers are differentiated by a phenyl-alkynyl bridge of increasing length at one meso-position, while for all the derivatives the two remaining opposite meso-positions are tailored with a phenyl moiety bearing a short polyether chain. Coordination of the four pyridyl groups with appropriate metal fragments may be exploited to construct tubular hollow structures, with varied internal sizes, depending on the choice of the porphyrin dimer component

TeHV3 outbreak characterization in captive Testudo spp.

Italian Tortoises species are considered either endangered or near threatened according to International Union for Conservation of Nature. When pet tortoises are abandoned or found injured or seized following illegal detention, they are sent to wildlife rehabilitation centers. From 2008, the Testudo spp. population housed in the WWF Vanzago’s oasis exhibited clinical signs compatible with Testudinid herpesviurs 3 (TeHV3) infection.  By the end of 2012 all Testudo had died. The presence of TeHV3 was investigated by molecular biology and pathology. All the tortoises housed in Vanzago resulted ELISA positive for the presence of anti-TeHV3 antibodies except one T. hermanni. Of these, 12 animals died and were all necropsied. Lesion frequency distribution was evaluate by histology. PCR was positive in 8/12 tortoises. To better complement the epidemiological evaluation of the virus in northern Italy, 20 retrospective cases were selected from the archive of the University of Milan. Of these, 5 were TeHV3 PCR positive. Lesions closely resembled those of the Vanzago’s population. These results are consistent with a high prevalence of TeHV3 in northern Italy. The finding of intranuclear inclusion bodies demonstrated to be specific but not sensitive. TeHV3 diagnostic pathological lesions have been reported to vary according with host immune response and by the viral replicative status. Molecular techniques were often necessary to confirm the infection. According to the literature and to our findings, T. hermanni spp. seems the species with higher mortality and lower antibody concentrations when infected with TeHV3.Italian Tortoises species are considered either endangered or near threatened according to International Union for Conservation of Nature. When pet tortoises are abandoned or found injured or seized following illegal detention, they are sent to wildlife rehabilitation centers. From 2008, the Testudo spp. population housed in the WWF Vanzago’s oasis exhibited clinical signs compatible with Testudinid herpesviurs 3 (TeHV3) infection.  By the end of 2012 all Testudo had died. The presence of TeHV3 was investigated by molecular biology and pathology. All the tortoises housed in Vanzago resulted ELISA positive for the presence of anti-TeHV3 antibodies except one T. hermanni. Of these, 12 animals died and were all necropsied. Lesion frequency distribution was evaluate by histology. PCR was positive in 8/12 tortoises. To better complement the epidemiological evaluation of the virus in northern Italy, 20 retrospective cases were selected from the archive of the University of Milan. Of these, 5 were TeHV3 PCR positive. Lesions closely resembled those of the Vanzago’s population. These results are consistent with a high prevalence of TeHV3 in northern Italy. The finding of intranuclear inclusion bodies demonstrated to be specific but not sensitive. TeHV3 diagnostic pathological lesions have been reported to vary according with host immune response and by the viral replicative status. Molecular techniques were often necessary to confirm the infection. According to the literature and to our findings, T. hermanni spp. seems the species with higher mortality and lower antibody concentrations when infected with TeHV3

Guanine-based amphiphiles: synthesis, ion transport properties and biological activity

Novel amphiphilic guanine derivatives, here named Gua1 and Gua2, have been prepared through few, simple and efficient synthetic steps. In ion transport experiments through phospholipid bilayers, carried out to evaluate their ability to mediate H(+) transport, Gua2 showed high activity. When this compound was investigated for ion-selective transport activities, no major differences were observed in the behaviour with cations while, in the case of anions, selective activity was observed in the series I(-)>Br(-)>Cl(-)>F(-). The bioactivity of these guanine analogues has been evaluated on a panel of human tumour and non-tumour cell lines in preliminary in vitro cytotoxicity assays, showing a relevant antiproliferative profile for Gua2

Evaluation of cytological diagnostic accuracy for canine splenic neoplasms : an investigation in 78 cases using STARD guidelines

Cytology represents a useful diagnostic tool in the preliminary clinical approach to canine splenic lesions, and may prevent unnecessary splenectomy. However, few studies have evaluated diagnostic accuracy of cytology in the diagnosis of canine splenic neoplasms. The aim of this study was to determine overall accuracy, sensitivity, specificity, positive and negative predictive values (i.e. diagnostic accuracy indexes) of cytology for canine splenic neoplasms following Standards for the Reporting of Diagnostic Accuracy Studies (STARD) guidelines. A consecutive series of canine splenic cytological samples was retrospectively retrieved from the database of the Diagnostic Pathology Service of the Department of Veterinary Medicine (DIMEVET\u2014University of Milan). Histopathology was set as the diagnostic reference standard. Cytological cases were enrolled when slides were available for review and when the same lesion was submitted for histopathology. Seventy-eight (78) lesions were included in the study. By histopathology, 56 were neoplastic and 22 were non-neoplastic. Cytology had an overall accuracy of 73.08% (95% C.I. 61.84%-82.50%), sensitivity of 64.29% (95% C.I. 50.36%-76.64%), specificity of 95.45% (95% C.I. 77.16%-99.88%), and positive and negative predictive values of 97.3% (95% C.I. 84.01%-99.60%) and 51.22% (95% C.I. 42.21%-60.15%), respectively. Low sensitivity and negative predictive value were balanced by very high specificity and positive predictive value. When positive for neoplasia, cytology represents a useful diagnostic tool to rule in splenic neoplasia, prompting surgery independently from other diagnostic tests. Conversely, a negative cytological result requires additional investigations to confirm the dog to be disease free