Stimulation of Activin A/Nodal signaling is insufficient to induce definitive endoderm formation of cord blood-derived unrestricted somatic stem cells

Introduction: Unrestricted somatic stem cells (USSC) derived from umbilical cord blood are an attractive alternative to human embryonic stem cells (hESC) for cellular therapy. USSC are capable of forming cells representative of all three germ line layers. The aim of this study was to determine the potential of USSC to form definitive endoderm following induction with Activin A, a protein known to specify definitive endoderm formation of hESC. Methods: USSC were cultured for (1) three days with or without 100 ng/ml Activin A in either serum-free, low-serum or serum-containing media, (2) three days with or without 100 ng/ml Activin A in combination with 10 ng/ml FGF4 in pre-induction medium, or (3) four days with or without small molecules Induce Definitive Endoderm (IDE1, 100 nM; IDE2, 200 nM) in serum-free media. Formation of definitive endoderm was assessed using RT-PCR for gene markers of endoderm (Sox17, FOXA2 and TTF1) and lung epithelium (surfactant protein C; SPC) and cystic fibrosis transmembrane conductance regulator; CFTR). The differentiation capacity of Activin A treated USSC was also assessed. Results: Activin A or IDE1/2 induced formation of Sox17+ definitive endoderm from hESC but not from USSC. Activin A treated USSC retained their capacity to form cells of the ectoderm (nerve), mesoderm (bone) and endoderm (lung). Activin A in combination with FGF4 did not induce formation of Sox17+ definitive endoderm from USSC. USSC express both Activin A receptor subunits at the mRNA and protein level, indicating that these cells are capable of binding Activin A. Conclusions: Stimulation of the Nodal signaling pathway with Activin A or IDE1/2 is insufficient to induce definitive endoderm formation from USSC, indicating that USSC differ in their stem cell potential from hESC

Arbovirus Screening in Mosquitoes in Emilia-Romagna (Italy, 2021) and Isolation of Tahyna Virus

Several viruses can be transmitted by mosquitoes. We searched some of these viruses in 20,778 mosquitoes, collected in 95 traps on the plains of Emilia-Romagna (North of Italy) in 2021. We detected West Nile virus (WNV) and Usutu virus (USUV) in pools of Culex (Cx.) pipiens. In addition, we detected two insect-specific flaviviruses in three pools of Aedes (Ae.) caspius and in two of Ae. vexans. Tahyna virus (TAHV) was detected in six pools, three of Ae. caspius and three of Cx. pipiens, and one isolated strain was obtained from one of the Ae. caspius pools. Moreover, we detected TAHV in pools of several mosquito species (Ae. caspius, Ae. vexans, Ae. albopictus, Anopheles maculipennis s.l.) collected in the previous year of surveillance. Our data indicate Ae. caspius as the species most infected with TAHV in the surveyed area. Together with the likely plasticity of the cycle, we reported strong genome stability of the TAHV, probably linked to a successful adaptation of the virus to its ecological niche. Interestingly, in six pools of Cx. pipiens we detected two associated viruses among USUV, WNV, TAHV and all the three viruses in two pools. This result allows us to assume the presence of particular conditions that prompt the circulation of arboviruses, creating the conditions for viral hot spots. While no human diseases related to Tahyna virus were reported in Italy, its detection over the years suggests that it is worth investigating this virus as a potential cause of disease in humans in order to assess its health burden. IMPORTANCE We reported in this work the detection of three Arboviruses (Arthropod-borne viruses) in mosquitoes collected in Emilia-Romagna in 2021. In addition to West Nile and Usutu viruses, which were reported from more than 10 years in the study area, we detected and isolated Tahyna virus (TAHV). We also reported detections of TAHV obtained in previous years of surveillance in different species of mosquitoes. TAHV is the potential causative agent of summer influenza-like diseases and also of meningitis. Even if human cases of disease referable to this virus are not reported in Italy, its relevant presence in mosquitoes suggests investigating the possibility they could

ORIGO: A mission concept to challenge planetesimal formation theories

Comets are generally considered among the most pristine objects in our Solar System. There have thus been significant efforts to understand these bodies. During the past decades, we have seen significant progress in our theoretical understanding of planetesimal/cometesimals (the precursors of comets) formation. Recent space missions—such as ESA’s Rosetta mission to comet 67P/Churyumov-Gerasimenko—have provided observations claimed by proponents of different comet formation theories to validate their scenarios. Yet, no single formation paradigm could be definitively proven. Given the importance of understanding how the first bodies in our Solar System formed, we propose a dedicated mission to address this issue. ORIGO will deliver a lander to the surface of a cometary nucleus where it will characterise the first five m of the subsurface. With remote sensing instruments and the deployment of payload into a borehole, we will be able to study the physico-chemical structure of ancient, unmodified material. The mission has been designed to fit into the ESA M-class mission budget

West Nile virus transmission. results from the integrated surveillance system in Italy, 2008 to 2015

IIn Italy a national Plan for the surveillance of imported and autochthonous human vector-borne diseases (chikungunya, dengue, Zika virus disease and West Nile virus (WNV) disease) that integrates human and veterinary (animals and vectors) surveillance, is issued and revised annually according with the observed epidemiological changes. Here we describe results of the WNV integrated veterinary and human surveillance systems in Italy from 2008 to 2015. A real time data exchange protocol is in place between the surveillance systems to rapidly identify occurrence of human and animal cases and to define and update the map of affected areas i.e. provinces during the vector activity period from June to October. WNV continues to cause severe illnesses in Italy during every transmission season, albeit cases are sporadic and the epidemiology varies by virus lineage and geographic area. The integration of surveillance activities and a multidisciplinary approach made it possible and have been fundamental in supporting implementation of and/or strengthening preventive measures aimed at reducing the risk of transmission of WNV trough blood, tissues and organ donation and to implementing further measures for vector control

Origo - an ESA M-class mission proposal to challenge planetesimal formation theories.

The Origo mission was submitted in response to the 2021 call for a Medium-size mission opportunity in ESA's Science Programme.The goal of Origo is to inform and challenge planetesimal formation theories. Understanding how planetesimals form in protoplanetary disks is arguably one of the biggest open questions in planetary science. To this end, it is indispensable to collect ground truths about the physico-chemical structure of the most pristine and undisturbed material available in our Solar System. Origo seeks to resolve the question of whether this icy material can still be found and thoroughly analysed in the sub-surface of comets.Specifically, Origo aims to address the following immediate science questions:Were cometesimals formed by distinct building blocks such as e.g. "pebbles", hierarchical sub-units, or fractal distributions? How did refractory and volatile materials come together during planetesimal growth e.g. did icy and refractory grains grow separately and come together later, or did refractory grains serve as condensation nuclei for volatiles? Did the building blocks of planetesimals all form in the vicinity of each other, or was there significant mixing of material within the protoplanetary disk? To answer these questions Origo will deliver a lander to a comet where we will characterise the first five meters of the subsurface with a combination of remote-sensing and payloads lowered into a borehole. Our instruments will examine the small scale physico-chemical structure. This approach will allow us to address the following objectives, each of which informs the respective science question: Reveal the existence of building blocks of a cometary nucleus from the (sub-)micron to metre scale by exploring unmodified material. Determine the physical structure of these building blocks, in particular, the size distribution of components and how refractory and volatile constituents are mixed and/or coupled. Characterise the composition of the building blocks by identifying and quantifying the major ices and refractory components. Over the past decade, significant theoretical advances have been achieved in working out possible planetesimal formation scenarios.The two leading hypotheses for how planetesimals formed from sub-micron dust and ice particles in the proto-planetary nebula can be classified into two groups:the hierarchical accretion of dust and ice grains to form planetesimals; and the growth of so-called pebbles, which are then brought to gentle gravitational collapse to form larger bodies by e.g. the streaming instability. These competing theories only have indirect proof from observations.Direct evidence, i.e. ground truths, about the building blocks of planetesimals remain hidden. Origo would challenge these theories by examining the physico-chemical structure of the most pristine material available in our Solar System. Though the proposal was not retained for step 2 we present our concept for community discussion

Rho/ROCK pathway is essential to the expansion, differentiation, and morphological rearrangements of human neural stem/progenitor cells induced by lysophosphatidic acid

We previously reported that lysophosphatidic acid (LPA) inhibits the neuronal differentiation of human embryonic stem cells (hESC). We extended these studies by analyzing LPA\u27s effects on the expansion of neural stem/ progenitor cells (NS/PC) derived from hESCs and human induced pluripotent stem cells (iPSC), and we assessed whether data obtained on the neural differentiation of hESCs were relevant to iPSCs. We showed that hESCs and iPSCs exhibited comparable mRNA expression profiles of LPA receptors and producing enzymes upon neural differentiation. We demonstrated that LPA inhibited the expansion of NS/PCs of both origins, mainly by increased apoptosis in a Rho/Rho-associated kinase (ROCK)-dependent mechanism. Furthermore, LPA inhibited the neuronal differentiation of iPSCs. Lastly, LPA induced neurite retraction of NS/ PC-derived early neurons through Rho/ROCK, which was accompanied by myosin light chain (MLC) phosphorylation. Our data demonstrate the consistency of LPA effects across various sources of human NS/PCs, rendering hESCs and iPSCs valuable models for studying lysophospholipid signaling in human neural cells. Our data also highlight the importance of the Rho/ROCK pathway in human NS/ PCs. As LPA levels are increased in the central nervous system (CNS) following injury, LPA-mediated effects on NS/ PCs and early neurons could contribute to the poor neurogenesis observed in the CNS following injury

If Human Brain Organoids Are the Answer to Understanding Dementia, What Are the Questions?

© The Author(s) 2020. Because our beliefs regarding our individuality, autonomy, and personhood are intimately bound up with our brains, there is a public fascination with cerebral organoids, the “mini-brain,” the “brain in a dish”. At the same time, the ethical issues around organoids are only now being explored. What are the prospects of using human cerebral organoids to better understand, treat, or prevent dementia? Will human organoids represent an improvement on the current, less-than-satisfactory, animal models? When considering these questions, two major issues arise. One is the general challenge associated with using any stem cell–generated preparation for in vitro modelling (challenges amplified when using organoids compared with simpler cell culture systems). The other relates to complexities associated with defining and understanding what we mean by the term “dementia.” We discuss 10 puzzles, issues, and stumbling blocks to watch for in the quest to model “dementia in a dish.