First report of Bufo bufo (Linnaeus, 1758) from Sardinia (Italy)

The Common toad Bufo bufo (Linneaus, 1758) was found for the first time in Sardinia in 2016 during herpetological surveys. The species appears to be well established in the finding area. Many adults and juveniles, tadpoles at different developmental stages, and eggs have been found during repeated monitoring. In order to infer the geographic origin of the Sardinian population, we amplified two mitochondrial markers (16S, cytb) and compared sequences with those available for the species across its natural range. We also screened samples for the presence of Bd pathogen to assess the risk of infection mediated by the species in the area. Results suggest that Sardinian individuals are genetically close to the Central Italian populations, although they show a unique distinct haplotype. Though the species should be considered allochthonous to the Island, further molecular and ecological data are urgently needed to assess the genetic structure and the possible impact on the local fauna, which is largely composed by endemic taxa. Particularly, possible interactions with other native amphibians like the green toad Bufo balearicus (Boettger, 1880), also present in the area, should be investigated, both in terms of competition for breeding sites and genetic pollution, as these species are already known to hybridize in the wild

Transmembrane protein TMEM230, regulator of metalloproteins and motor proteins in gliomas and gliosis

Glial cells provide physical and chemical support and protection for neurons and for the extracellular compartments of neural tissue through secretion of soluble factors, insoluble scaffolds, and vesicles. Additionally, glial cells have regenerative capacity by remodeling their physical microenvironment and changing physiological properties of diverse cell types in their proximity. Various types of aberrant glial and macrophage cells are associated with human diseases, disorders, and malignancy. We previously demonstrated that transmembrane protein, TMEM230 has tissue revascularization and regenerating capacity by its ability to secrete pro-angiogenic factors and metalloproteinases, inducing endothelial cell sprouting and channel formation. In healthy normal neural tissue, TMEM230 is predominantly expressed in glial and marcophate cells, suggesting a prominent role in neural tissue homeostasis. TMEM230 regulation of the endomembrane system was supported by co-expression with RNASET2 (lysosome, mitochondria, and vesicles) and STEAP family members (Golgi complex). Intracellular trafficking and extracellular secretion of glial cellular components are associated with endocytosis, exocytosis and phagocytosis mediated by motor proteins. Trafficked components include metalloproteins, metalloproteinases, glycans, and glycoconjugate processing and digesting enzymes that function in phagosomes and vesicles to regulate normal neural tissue microenvironment, homeostasis, stress response, and repair following neural tissue injury or degeneration. Aberrantly high sustained levels TMEM230 promotes metalloprotein expression, trafficking and secretion which contribute to tumor associated infiltration and hypervascularization of high tumor grade gliomas. Following injury of the central nervous or peripheral systems, transcient regulated upregulation of TMEM230 promotes tissue wound healing, remodeling and revascularization by activating glial and macrophage generated microchannels/microtubules (referred to as vascular mimicry) and blood vessel sprouting and branching. Our results support that TMEM230 may act as a master regulator of motor protein mediated trafficking and compartmentalization of a large class of metalloproteins in gliomas and gliosis

Lack of SARS-CoV-2 RNA environmental contamination in a tertiary referral hospital for infectious diseases in Northern Italy

none140noNAnoneColaneri M.; Seminari E.; Piralla A.; Zuccaro V.; Di Filippo A.; Baldanti F.; Bruno R.; Mondelli M.U.; Brunetti E.; Di Matteo A.; Maiocchi L.; Pagnucco L.; Mariani B.; Ludovisi S.; Lissandrin R.; Parisi A.; Sacchi P.; Patruno S.F.A.; Michelone G.; Gulminetti R.; Zanaboni D.; Novati S.; Maserati R.; Orsolini P.; Vecchia M.; Sciarra M.; Asperges E.; Sambo M.; Biscarini S.; Lupi M.; Roda S.; Chiara Pieri T.; Gallazzi I.; Sachs M.; Valsecchi P.; Perlini S.; Alfano C.; Bonzano M.; Briganti F.; Crescenzi G.; Giulia Falchi A.; Guarnone R.; Guglielmana B.; Maggi E.; Martino I.; Pettenazza P.; Pioli di Marco S.; Quaglia F.; Sabena A.; Salinaro F.; Speciale F.; Zunino I.; De Lorenzo M.; Secco G.; Dimitry L.; Cappa G.; Maisak I.; Chiodi B.; Sciarrini M.; Barcella B.; Resta F.; Moroni L.; Vezzoni G.; Scattaglia L.; Boscolo E.; Zattera C.; Michele Fidel T.; Vincenzo C.; Vignaroli D.; Bazzini M.; Iotti G.; Mojoli F.; Belliato M.; Perotti L.; Mongodi S.; Tavazzi G.; Marseglia G.; Licari A.; Brambilla I.; Daniela B.; Antonella B.; Patrizia C.; Giulia C.; Giuditta C.; Marta C.; Rossana D.; Milena F.; Bianca M.; Roberta M.; Enza M.; Stefania P.; Maurizio P.; Elena P.; Antonio P.; Francesca R.; Antonella S.; Maurizio Z.; Guy A.; Laura B.; Ermanna C.; Giuliana C.; Luca D.; Gabriella F.; Gabriella G.; Alessia G.; Viviana L.; Claudia L.; Valentina M.; Simona P.; Marta P.; Alice B.; Giacomo C.; Irene C.; Alfonso C.; Di Martino R.; Di Napoli A.; Alessandro F.; Guglielmo F.; Loretta F.; Federica G.; Alessandra M.; Federica N.; Giacomo R.; Beatrice R.; Maria S.I.; Monica T.; Nepita Edoardo V.; Calvi M.; Tizzoni M.; Nicora C.; Triarico A.; Petronella V.; Marena C.; Muzzi A.; Lago P.; Comandatore F.; Bissignandi G.; Gaiarsa S.; Rettani M.; Bandi C.Colaneri, M.; Seminari, E.; Piralla, A.; Zuccaro, V.; Di Filippo, A.; Baldanti, F.; Bruno, R.; Mondelli, M. U.; Brunetti, E.; Di Matteo, A.; Maiocchi, L.; Pagnucco, L.; Mariani, B.; Ludovisi, S.; Lissandrin, R.; Parisi, A.; Sacchi, P.; Patruno, S. F. A.; Michelone, G.; Gulminetti, R.; Zanaboni, D.; Novati, S.; Maserati, R.; Orsolini, P.; Vecchia, M.; Sciarra, M.; Asperges, E.; Sambo, M.; Biscarini, S.; Lupi, M.; Roda, S.; Chiara Pieri, T.; Gallazzi, I.; Sachs, M.; Valsecchi, P.; Perlini, S.; Alfano, C.; Bonzano, M.; Briganti, F.; Crescenzi, G.; Giulia Falchi, A.; Guarnone, R.; Guglielmana, B.; Maggi, E.; Martino, I.; Pettenazza, P.; Pioli di Marco, S.; Quaglia, F.; Sabena, A.; Salinaro, F.; Speciale, F.; Zunino, I.; De Lorenzo, M.; Secco, G.; Dimitry, L.; Cappa, G.; Maisak, I.; Chiodi, B.; Sciarrini, M.; Barcella, B.; Resta, F.; Moroni, L.; Vezzoni, G.; Scattaglia, L.; Boscolo, E.; Zattera, C.; Michele Fidel, T.; Vincenzo, C.; Vignaroli, D.; Bazzini, M.; Iotti, G.; Mojoli, F.; Belliato, M.; Perotti, L.; Mongodi, S.; Tavazzi, G.; Marseglia, G.; Licari, A.; Brambilla, I.; Daniela, B.; Antonella, B.; Patrizia, C.; Giulia, C.; Giuditta, C.; Marta, C.; D'Alterio, Rossana; Milena, F.; Bianca, M.; Roberta, M.; Enza, M.; Stefania, P.; Maurizio, P.; Elena, P.; Antonio, P.; Francesca, R.; Antonella, S.; Maurizio, Z.; Guy, A.; Laura, B.; Ermanna, C.; Giuliana, C.; Luca, D.; Gabriella, F.; Gabriella, G.; Alessia, G.; Viviana, L.; Meisina, Claudia; Valentina, M.; Simona, P.; Marta, P.; Alice, B.; Giacomo, C.; Irene, C.; Alfonso, C.; Di Martino, R.; Di Napoli, A.; Alessandro, F.; Guglielmo, F.; Loretta, F.; Federica, G.; Albertini, Alessandra; Federica, N.; Giacomo, R.; Beatrice, R.; Maria, S. I.; Monica, T.; Nepita Edoardo, V.; Calvi, M.; Tizzoni, M.; Nicora, C.; Triarico, A.; Petronella, V.; Marena, C.; Muzzi, A.; Lago, P.; Comandatore, F.; Bissignandi, G.; Gaiarsa, S.; Rettani, M.; Bandi, C

Clinical characteristics of coronavirus disease (COVID-19) early findings from a teaching hospital in Pavia, North Italy, 21 to 28 February 2020

We describe clinical characteristics, treatments and outcomes of 44 Caucasian patients with coronavirus disease (COVID-19) at a single hospital in Pavia, Italy, from 21\u201328 February 2020, at the beginning of the outbreak in Europe. Seventeen patients developed severe disease, two died. After a median of 6 days, 14 patients were discharged from hospital. Predictors of lower odds of discharge were age>65 years, antiviral treatment and for severe disease, lactate dehydrogenase >300 mg/dL

Molecular markers and bioinformatics in species detection: two case studies in Pelophylax spp. (Amphibia, Ranidae) and in a novel bacterial endosymbiont, in ciliate protista

Species detection is one of the older problems encountered by science. This detection implicitly needs the identification of several features that are peculiar of a group, or taxa, such that their cumulative presence or absence defines the category of the target. Initially, taxonomies were based on morphological characters, with some limitations as there are species lacking the needed informative features, as prokaryotic or cryptic species. From the discovery of genotype, sequences have proven as powerful features to be used to infer taxonomy and phylogenetic trees. A sample can be affiliated to a taxonomy by finding a homology to an already taxonomically classified sequence present in a public database. However, if the target sequence does not have a match, then a phylogenetic tree constructed together with the found similar sequences can be used to infer their relationships. This type of taxonomy annotation can be challenging as there is not a consensus about how much distant two taxa have to be to be considered of different species. In this PhD thesis I consider two taxonomically challenging case studies, the first involving a complex of morphologically cryptic interbreeding Anuran species, the second a bacterial endosymbiont of a ciliate protozoa. The first case is focused on Pelophylax, a genus of morphologically cryptic anuran species that show a form of sexual parasitism, called hybridogenesis, where inter-species mate produces viable offspring. Some species can be identified on a bio-acoustic basis, but the presence of hybrids makes the detection unaffordable. The identification is more precisely performed using mitochondrial DNA (mtDNA) and Short Tandem Repeats (STR) markers. I coupled these two techniques to classify animals sampled in the Po Valley, near Pavia, where two autochthonous taxa can be found together with allochthonous. A correct detection of these species is important to assess the conservation status of the local taxa and the impact of the allochthonous ones. The second case is focused on the study if a prokaryotic endosymbiont of a ciliate Protista, where morphological traits are virtually useless to determinate the taxonomy. Due to differences between host and symbionts, it is possible to use molecular traits to discern the species, as using 16s rRNA for Bacteria and 18s rRNA for Eukaryotes. However, this approach may only provide species identification, not genomic information, which is needed to get a functional understanding of the symbiotic system, as also of the endosymbiotic species, which may be unknown. With this purpose, both organisms are sequenced together by using Whole Genome Sequencing (WGS) with Next Generation Sequencing (NGS) technologies. However, this procedure allows to obtain portion of the genomes fragmented into different contigs, which than have to be deconvolved to obtain separate genomes. We then decided to develop a fully automated tool, called SeqDex, able to deconvolve host-endosymbiont dataset by coupling partial taxonomic affiliations (homology derived) to composition analysis to predict the affiliations of all the sequences 4 using state of the art machine learning algorithms. The second case study is composed by three Spirostomum samples, which showed evidence of presence of a Neisseriales bacterium inside the ciliate cells. I have used SeqDex to deconvolve this dataset to reconstruct partially the endosymbionts genomes and perform functional analysis to infer their role and the nature of the relationship that bound the hosts and the bacteria

First report of bufo bufo (Linnaeus, 1758) from sardinia (Italy)

The Common toad Bufo bufo (Linneaus, 1758) was found for the first time in Sardinia in 2016 during her-petological surveys. The species appears to be well established in the finding area. Many adults and juveniles, tadpoles at different developmental stages, and eggs have been found during repeated monitoring. In order to infer the geographic origin of the Sardinian population, we amplified two mitochondrial markers (16S, cytb) and compared sequences with those available for the species across its natural range. We also screened samples for the presence of Bd pathogen to assess the risk of infection mediated by the species in the area. Results suggest that Sardinian individuals are genetically close to the Central Italian populations, although they show a unique distinct haplotype. Though the species should be considered allochthonous to the Island, further molecular and ecological data are urgently needed to assess the genetic structure and the possible impact on the local fauna, which is largely composed by endemic taxa. Particularly, possible interactions with other native amphibians like the green toad Bufo balearicus (Boettger, 1880), also present in the area, should be investigated, both in terms of competition for breeding sites and genetic pollution, as these species are already known to hybridize in the wild

First report of <em>Bufo bufo</em> (Linnaeus, 1758) from Sardinia (Italy)

The Common toad Bufo bufo (Linneaus, 1758) was found for the first time in Sardinia in 2016 during herpetological surveys. The species appears to be well established in the finding area. Many adults and juveniles, tadpoles at different developmental stages, and eggs have been found during repeated monitoring. In order to infer the geographic origin of the Sardinian population, we amplified two mitochondrial markers (16S, cytb) and compared sequences with those available for the species across its natural range. We also screened samples for the presence of Bd pathogen to assess the risk of infection mediated by the species in the area. Results suggest that Sardinian individuals are genetically close to the Central Italian populations, although they show a unique distinct haplotype. Though the species should be considered allochthonous to the Island, further molecular and ecological data are urgently needed to assess the genetic structure and the possible impact on the local fauna, which is largely composed by endemic taxa. Particularly, possible interactions with other native amphibians like the green toad Bufo balearicus (Boettger, 1880), also present in the area, should be investigated, both in terms of competition for breeding sites and genetic pollution, as these species are already known to hybridize in the wild

Absence of 6’Sialyllactose during lactation impairs cognitive capabilities and modulates gene expression

Aims: Human milk is the ideal source of nutrition for the new-born, promoting the development of cognitive capabilities. However, which components of maternal milk are involved in the proper development of executive functions has remained elusive. We hypothesized that the maturation of attention, cognitive flexibility, and memory depends on the neonatal bioavailability of a specific human milk oligosaccharide (HMOs), sialyl(alpha2,6)lactose (6’SL). Methods: To test this hypothesis, we evaluated the aforementioned cognitive capabilities in adult mice that received maternal milk containing different concentrations of 6’SL. To modulate the availability of 6’SL during lactation, we leveraged a genetically engineered mouse (C57BL/6-St6gal1tm1.1Jxm/J, KO), that provides milk without 6‘SL. Specifically, we performed a cross-fostering study in which wild-type (WT) mice were reared to either KO or WT dams. During lactation, maternal behaviour was analysed for potential effects of maternal care on offspring. Adult subjects were tested for spatial memory, working memory and sensorimotor gating. To understand the underpinning molecular mechanisms of potential effect, we performed an RNA-seq analysis on PFC and hippocampal samples. Furthermore, we addressed ex vivo long-term potentiation (LTP, an electrophysiological correlate of memory performance) in hippocampal slices. Results: Mice that received 6’SL deficient milk showed an impoverishment of spatial reference memory, working memory and attentional capabilities compared to control. Furthermore, this group showed an altered regulation in the expression of genes involved in PFC development and an alteration in LTP. Conclusions: These findings show that absence of 6’SL in maternal milk impairs cognitive functions, such as memory and attention

Exposure to 3&prime;Sialyllactose-Poor Milk during Lactation Impairs Cognitive Capabilities in Adulthood

Breast milk exerts pivotal regulatory functions early in development whereby it contributes to the maturation of brain and associated cognitive functions. However, the specific components of maternal milk mediating this process have remained elusive. Sialylated human milk oligosaccharides (HMOs) represent likely candidates since they constitute the principal neonatal dietary source of sialic acid, which is crucial for brain development and neuronal patterning. We hypothesize that the selective neonatal lactational deprivation of a specific sialylated HMOs, sialyl(alpha2,3)lactose (3&prime;SL), may impair cognitive capabilities (attention, cognitive flexibility, and memory) in adulthood in a preclinical model. To operationalize this hypothesis, we cross-fostered wild-type (WT) mouse pups to B6.129-St3gal4tm1.1Jxm/J dams, knock-out (KO) for the gene synthesizing 3&prime;SL, thereby providing milk with approximately 80% 3&prime;SL content reduction. We thus exposed lactating WT pups to a selective reduction of 3&prime;SL and investigated multiple cognitive domains (including memory and attention) in adulthood. Furthermore, to account for the underlying electrophysiological correlates, we investigated hippocampal long-term potentiation (LTP). Neonatal access to 3&prime;SL-poor milk resulted in decreased attention, spatial and working memory, and altered LTP compared to the control group. These results support the hypothesis that early-life dietary sialylated HMOs exert a long-lasting role in the development of cognitive functions