Antiangiogenic resveratrol analogues by mild m-CPBA aromatic hydroxylation of 3,5-dimethoxystilbenes.

A mild treatment of the resveratrol analogue 3,5,4′-trimethoxystilbene 2 with m-CPBA afforded two hydroxylated methoxystilbenes 5 and 6 by direct aromatic hydroxylation. A similar protocol was applied to other stilbenes bearing a 3,5-dimethoxy moiety, namely tetramethoxystilbenes 7 and 10 to obtain respectively the hydroxylated analogues 8, 9 and 11, 12. The substrate 2 and the new compounds 5, 8 and 11 were evaluated as anti-angiogenic agents and proved significantly active in the range 1 – 100 μM

A relevant long-term impact of the circulation of a potentially contaminated vaccine on the distribution of scrapie in Italy. Results from a retrospective cohort study

A sudden increase in the incidence of scrapie in Italy in 1997 was subsequently linked to the use of a potentially infected vaccine against contagious agalactia. The relative risk for the exposed farms ranged between 6 and 40. The aim of this study was to assess the long-term impact of exposure to the potentially scrapie-contaminated vaccine on the Italian classical scrapie epidemic. We carried out a retrospective cohort study, fitting mixed-effects Poisson regression models, dividing national geographic areas into exposure categories on the basis of the vaccine circulation levels. We took into account the sensitivity of the surveillance system applied in the different areas. The population attributable fraction (PAF) was used to assess the impact on the total population of farms associated with the effect of circulation of the vaccine. The provinces where the vaccine was more often sold were noted to have a higher level of disease when compared to those provinces where the vaccine was sold less often (incidence rate ratio [IRR]: 2.7; 95% confidence interval [CI]: 1.1-6.5). The population attributable fraction was high (68.4%). Standardization techniques allowed to account for the potential of geographical variability in the sensitivity of the Italian surveillance system. Although the number of the directly exposed farms was limited, an important long-term impact of the vaccine circulation could be quantified in terms of secondary outbreaks likely due to the exchange of animals from directly exposed flocks

Evolution of Swine Influenza Virus H3N2 in Vaccinated and Nonvaccinated Pigs after Previous Natural H1N1 Infection

Swine influenza viruses (SIV) produce a highly contagious and worldwide distributed disease that can cause important economic losses to the pig industry. Currently, this virus is endemic in farms and, although used limitedly, trivalent vaccine application is the most extended strategy to control SIV. The presence of pre-existing immunity against SIV may modulate the evolutionary dynamic of this virus. To better understand these dynamics, the viral variants generated in vaccinated and nonvaccinated H3N2 challenged pigs after recovery from a natural A(H1N1) pdm09 infection were determined and analyzed. In total, seventeen whole SIV genomes were determined, 6 from vaccinated, and 10 from nonvaccinated animals and their inoculum, by NGS. Herein, 214 de novo substitutions were found along all SIV segments, 44 of them being nonsynonymous ones with an allele frequency greater than 5%. Nonsynonymous substitutions were not found in NP; meanwhile, many of these were allocated in PB2, PB1, and NS1 proteins. Regarding HA and NA proteins, higher nucleotide diversity, proportionally more nonsynonymous substitutions with an allele frequency greater than 5%, and different domain allocations of mutants, were observed in vaccinated animals, indicating different evolutionary dynamics. This study highlights the rapid adaptability of SIV in different environments.info:eu-repo/semantics/publishedVersio

Identification and Characterization of Swine Influenza Virus H1N1 Variants Generated in Vaccinated and Nonvaccinated, Challenged Pigs

Influenza viruses represent a continuous threat to both animal and human health. The 2009 H1N1 A influenza pandemic highlighted the importance of a swine host in the adaptation of influenza viruses to humans. Nowadays, one of the most extended strategies used to control swine influenza viruses (SIVs) is the trivalent vaccine application, whose formulation contains the most frequently circulating SIV subtypes H1N1, H1N2, and H3N2. These vaccines do not provide full protection against the virus, allowing its replication, evolution, and adaptation. To better understand the main mechanisms that shape viral evolution, here, the SIV intra-host diversity was analyzed in samples collected from both vaccinated and nonvaccinated animals challenged with the H1N1 influenza A virus. Twenty-eight whole SIV genomes were obtained by next-generation sequencing, and differences in nucleotide variants between groups were established. Substitutions were allocated along all influenza genetic segments, while the most relevant nonsynonymous substitutions were allocated in the NS1 protein on samples collected from vaccinated animals, suggesting that SIV is continuously evolving despite vaccine application. Moreover, new viral variants were found in both vaccinated and nonvaccinated pigs, showing relevant substitutions in the HA, NA, and NP proteins, which may increase viral fitness under field conditions.info:eu-repo/semantics/publishedVersio

Reconstructing the recent West Nile virus lineage 2 epidemic in Europe and Italy using discrete and continuous phylogeography

West Nile virus lineage 2 (WNV-2) was mainly confined to sub-Saharan Africa until the early 2000s, when it was identified for the first time in Central Europe causing outbreaks of human and animal infection. The aim of this study was to reconstruct the origin and dispersion of WNV-2 in Central Europe and Italy on a phylodynamic and phylogeographical basis. To this aim, discrete and continuous space phylogeographical models were applied to a total of 33 newly characterised full-length viral genomes obtained from mosquitoes, birds and humans in Northern Italy in the years 2013-2015 aligned with 64 complete sequences isolated mainly in Europe. The European isolates segregated into two highly significant clades: a small one including three sequences and a large clade including the majority of isolates obtained in Central Europe since 2004. Discrete phylogeographical analysis showed that the most probable location of the root of the largest European clade was in Hungary a mean 12.78 years ago. The European clade bifurcated into two highly supported subclades: one including most of the Central/East European isolates and the other encompassing all of the isolates obtained in Greece. The continuous space phylogeographical analysis of the Italian clade showed that WNV-2 entered Italy in about 2008, probably by crossing the Adriatic sea and reaching a central area of the Po Valley. The epidemic then spread simultaneously eastward, to reach the region of the Po delta in 2013, and westward to the border area between Lombardy and Piedmont in 2014; later, the western strain changed direction southward, and reached the central area of the Po valley once again in 2015. Over a period of about seven years, the virus spread all over an area of northern Italy by following the Po river and its main tributaries

Evolution of Swine Influenza Virus H3N2 in Vaccinated and Nonvaccinated Pigs after Previous Natural H1N1 Infection

Swine influenza viruses (SIV) produce a highly contagious and worldwide distributed disease that can cause important economic losses to the pig industry. Currently, this virus is endemic in farms and, although used limitedly, trivalent vaccine application is the most extended strategy to control SIV. The presence of pre-existing immunity against SIV may modulate the evolutionary dynamic of this virus. To better understand these dynamics, the viral variants generated in vaccinated and nonvaccinated H3N2 challenged pigs after recovery from a natural A(H1N1) pdm09 infection were determined and analyzed. In total, seventeen whole SIV genomes were determined, 6 from vaccinated, and 10 from nonvaccinated animals and their inoculum, by NGS. Herein, 214 de novo substitutions were found along all SIV segments, 44 of them being nonsynonymous ones with an allele frequency greater than 5%. Nonsynonymous substitutions were not found in NP; meanwhile, many of these were allocated in PB2, PB1, and NS1 proteins. Regarding HA and NA proteins, higher nucleotide diversity, proportionally more nonsynonymous substitutions with an allele frequency greater than 5%, and different domain allocations of mutants, were observed in vaccinated animals, indicating different evolutionary dynamics. This study highlights the rapid adaptability of SIV in different environments

Arcobacter spp. in raw milk from vending machines in Piedmont and occurrence of virulence genes in isolates

Arcobacter spp. has been recognized as an emerging foodborne pathogen and a hazard to human health. In the dairy chain, it has been isolated from different sources, nevertheless data on Arcobacter occurrence in raw milk provided by vending machines are few. This study aimed to identify potentially pathogenic Arcobacter spp. in raw milk intended for human consumption sold through vending machines located in Piedmont. In an 8-month period, 37 raw milk samples were collected from 24 dairy farms: 12 (32,4%) were collected directly in farm from bulk tank milk and 25 (67,6%) from vending machines. Eight (21,6%) out of the 37 milk samples and 7 (29,2%) out of the 24 dairy farms were positive for Arcobacter spp. by culture examination. Four (16%) out of the 25 samples from vending machines and 4 (33,3%) out of the 12 samples from bulk tank milk were positive. All 8 isolates were identified as A. butzleri both by MALDI-TOF MS and multiplex end-point PCR. According to the detection of virulence genes, a total of four Patho-types were highlighted: 5 isolates in P-type 1 and only one isolate for each of the P-types 2-3-4. A. butzleri isolates carrying encoding virulence factors genes were isolated from raw milk intended for human consumption: these findings strengthen the compulsory consumption after boiling as required by current legislation and suggest the need of enlarging the analytical investigations to other microorganisms not yet included in the food safety criteria

Physiological control of angiogenesis in the ovarian follicle and potential interferents

La funzionalità del follicolo ovarico è dipendente dall’azione di numerosi fattori che ne regolano l’attività agendo sulle cellule che lo compongono. Fra queste un ruolo chiave è svolto dalle cellule della granulosa. Esse infatti intervengono in diversi processi e, fra questi, quello angiogenico occupa un posto di rilievo. Gli studi sperimentali descritti in questa tesi sono stati condotti utilizzando come modello il follicolo antrale suino ed impiegando un saggio biologico di angiogenesi in vitro. Lo scopo è stato quello di indagare il ruolo svolto nella regolazione fisiologica dell’angiogenesi follicolare di tre fattori proteici, la Stanniocalcina-1, la Netrina-1, e Prolattina e, di contro, la potenziale azione interferente sul processo fisiologico di due sostanze esogene, ma potenzialmente in grado di raggiungere il follicolo, il Gossipolo e il Bisfenolo A. Inoltre, questo sistema sperimentale è stato utilizzato per indagare le propriètà biologiche di analoghi di sintesi del Resveratrolo e dei Lignani, potenziali agenti terapeutici per l’angiogenesi patologica. I risultati ottenuti hanno evidenziato che la Stanniocalcina-1, la Netrina-1 e la Prolattina rappresentano potenziali regolatori fisiologici locali dell’angiogenesi follicolare. Per quanto riguarda il Gossipolo e il Bisfenolo A, modulando l’attività funzionale delle cellule della granulosa, potrebbero causare un’alterazione del controllo fisiologico dei processi follicolari. Infine, gli analoghi di sintesi del Resveratrolo e dei Lignani hanno mostrato proprietà biologiche di estremo interesse nell’ottica di un loro potenziale utilizzo a scopo terapeutico.Ovarian follicle function is dependent on many factors that act on its cells to regulate their activities. In particular, a pivotal role is played by granulosa cells. Indeed, they are involved in different processes and, among these, angiogenesis have an important role. The experimental studies described in this thesis were conducted employing as a model the swine antral follicle and an in vitro angiogenesis bioassay. The aim was to investigate the role of proteins namely Stanniocalcin-1, Netrin-1, and Prolactin in the physiological regulation of follicular angiogenesis and, conversely, the potential interfering action on the physiological process of Gossypol and Bisphenol A, exogenous compounds virtually able to reach the follicle. In addition, this experimental system was used to investigate the biological functions of synthetic analogues of resveratrol and lignans, promosing therapeutic agents for pathological angiogenesis. The results showed that Stanniocalcin-1, Netrin-1 and Prolactin are likely local physiological regulators of follicular angiogenesis. As for Gossypol and Bisphenol A, they might change the physiological regulation of follicular processes modulating granulosa cells function. Finally, synthetic analogues of resveratrol and lignans have shown biological properties of great interest in view of their future therapeutic use

Gossypol, a polyphenolic aldehyde from cotton plant, interferes with swine granulosa cell function

Gossypol is a polyphenol isolated from the seed, roots and stem of cotton plant (Gossypium sp.) It has been associated with adverse effects on female reproduction, but recently also shown having promising effects against several malignancies. Its mechanisms of action are however still not fully understood. This study was therefore conducted to investigate the effect of 5 or 25 microg/mL gossypol on swine granulosa cell steroidogenic activity, redox status and Vascular Endothelial Growth Factor (VEGF) production. Study demonstrated that gossypol significantly (P<0.001) inhibited granulosa cell estradiol 17beta and progesterone production, an effect that could be at least partially mediated by an increase (P<0.05) of nitric oxide and superoxide anion production as a consequence of superoxide dismutase inhibition. Moreover, gossypol stimulates (P<0.001) VEGF production. In conclusion, study has demonstrated effects of gossypol on swine granulosa cell function in vitro. Effects on female swine fertility can not be exclude