Direct conversion of human pluripotent stem cells into cranial motor neurons using a piggyBac vector

Human pluripotent stem cells (PSCs) are widely used for in vitro disease modeling. One of the challenges in the field is represented by the ability of converting human PSCs into specific disease-relevant cell types. The nervous system is composed of a wide variety of neuronal types with selective vulnerability in neurodegenerative diseases. This is particularly relevant for motor neuron diseases, in which different motor neurons populations show a different susceptibility to degeneration. Here we developed a fast and efficient method to convert human induced Pluripotent Stem Cells into cranial motor neurons of the branchiomotor and visceral motor subtype. These populations represent the motor neuron subgroup that is primarily affected by a severe form of amyotrophic lateral sclerosis with bulbar onset and worst prognosis. This goal was achieved by stable integration of an inducible vector, based on the piggyBac transposon, allowing controlled activation of Ngn2, Isl1 and Phox2a (NIP). The NIP module effectively produced electrophysiologically active cranial motor neurons. Our method can be easily extended to PSCs carrying disease-associated mutations, thus providing a useful tool to shed light on the cellular and molecular bases of selective motor neuron vulnerability in pathological conditions

FUS mutant human motoneurons display altered transcriptome and microRNA pathways with implications for ALS pathogenesis

The FUS gene has been linked to amyotrophic lateral sclerosis (ALS). FUS is a ubiquitous RNA-binding protein, and the mechanisms leading to selective motoneuron loss downstream of ALS-linked mutations are largely unknown. We report the transcriptome analysis of human purified motoneurons, obtained from FUS wild-type or mutant isogenic induced pluripotent stem cells (iPSCs). Gene ontology analysis of differentially expressed genes identified significant enrichment of pathways previously associated to sporadic ALS and other neurological diseases. Several microRNAs (miRNAs) were also deregulated in FUS mutant motoneurons, including miR-375, involved in motoneuron survival. We report that relevant targets of miR-375, including the neural RNA-binding protein ELAVL4 and apoptotic factors, are aberrantly increased in FUS mutant motoneurons. Characterization of transcriptome changes in the cell type primarily affected by the disease contributes to the definition of the pathogenic mechanisms of FUS-linked ALS

Lessons from April 6, 2009 L'Aquila earthquake to enhance microzoning studies in near-field urban areas

AbstractThis study focuses on two weak points of the present procedure to carry out microzoning study in near-field areas: (1) the Ground Motion Prediction Equations (GMPEs), commonly used in the reference seismic hazard (RSH) assessment; (2) the ambient noise measurements to define the natural frequency of the near surface soils and the bedrock depth. The limitations of these approaches will be discussed throughout the paper based on the worldwide and Italian experiences performed after the 2009 L'Aquila earthquake and then confirmed by the most recent 2012 Emilia Romagna earthquake and the 2016–17 Central Italy seismic sequence. The critical issues faced are (A) the high variability of peak ground acceleration (PGA) values within the first 20–30 km far from the source which are not robustly interpolated by the GMPEs, (B) at the level 1 microzoning activity, the soil seismic response under strong motion shaking is characterized by microtremors' horizontal to vertical spectral ratios (HVSR) according to Nakamura's method. This latter technique is commonly applied not being fully compliant with the rules fixed by European scientists in 2004, after a 3-year project named Site EffectS assessment using AMbient Excitations (SESAME). Hereinafter, some "best practices" from recent Italian and International experiences of seismic hazard estimation and microzonation studies are reported in order to put forward two proposals: (a) to formulate site-specific GMPEs in near-field areas in terms of PGA and (b) to record microtremor measurements following accurately the SESAME advice in order to get robust and repeatable HVSR values and to limit their use to those geological contests that are actually horizontally layered

A New Approach in the Management of Triangular Fossa Auricular Defects: The Posterior Auricular Artery Perforator Antihelix-Conchal Flap (PAAP Flap).

Background: The reconstruction of extensive scapha or triangular fossa defects to the ear still represents a challenge considering the lack of local simple surgical procedures. Objective: To present a new perforator flap which is a modification of the Elsahy and Ladocsi chondrocutaneous rotation flaps. Patients and Methods: Ten patients, in whom the oncological resection to the triangular fossa did not include cartilage, but skin and perichondrium. Defects no larger than 3 cm in diameter were treated. The dissection of the flap was carried out from lateral to medial on the anterior subperichondral plane. The flap was thus raised without the cartilage component but with the anterior perichondrium branched with the perforating vessels. Results: All flaps survived and treated patients presented good aesthetic results. Conclusions: We think that this procedure represents a valuable option to be considered in reconstructing well selected composite defects to upper ear

FUS affects circular RNA expression in murine embryonic stem cell-derived motor neurons

The RNA-binding protein FUS participates in several RNA biosynthetic processes and has been linked to the pathogenesis of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia. Here we report that FUS controls back-splicing reactions leading to circular RNA (circRNA) production. We identified circRNAs expressed in in vitro -derived mouse motor neurons (MNs) and determined that the production of a considerable number of these circRNAs is regulated by FUS. Using RNAi and overexpression of wild-type and ALS-asso- ciated FUS mutants, we directly correlate the modulation of circRNA biogenesis with alteration of FUS nuclear levels and with putative toxic gain of function activities. We also demonstrate that FUS regulates circRNA biogenesis by binding the introns flanking the back-splicing junctions and that this control can be reproduced with artificial constructs. Most circRNAs are conserved in humans and specific ones are deregulated in human-induced pluripotent stem cell-derived MNs carrying the FUS P525L mutation associated with AL

Thu0349 autologous fat grafting in the treatment of patients with systemic sclerosis: current experience and future prospects

none13nomixedSpinella, Amelia; Pignatti, Marco; Citriniti, Giorgia; Lumetti, Federica; Cocchiara, Emanuele; Palermo, Adalgisa; Sighinolfi, Gianluca; Pacchioni, Lucrezia; Zaccaria, Giovanna; Lusetti, Irene Laura; Santis, Giorgio De; Salvarani, Carlo; Giuggioli, DiliaSpinella, Amelia; Pignatti, Marco; Citriniti, Giorgia; Lumetti, Federica; Cocchiara, Emanuele; Palermo, Adalgisa; Sighinolfi, Gianluca; Pacchioni, Lucrezia; Zaccaria, Giovanna; Lusetti, Irene Laura; Santis, Giorgio De; Salvarani, Carlo; Giuggioli, Dili

環境管理計画への市民参加とその規定因としてのエンパワーメント

<p>DMPC/2/siRNA treatment of Jurkat T cells, 48 hours from the beginning of transfection.</p

Health and nutrition studies related to cereal biodiversity: a participatory multi-actor literature review approach

Recently, a large and growing body of literature has investigated the health potential of different wheat species. In particular, a considerable number of studies dealing with nutritional aspects has grown up around the theme of the recovery of ancient wheat varieties (species that have remained unchanged over the last hundred years). According to several studies, indeed, ancient varieties present a healthier nutritional profile than modern ones. In the framework of the European project "CERERE, CEreal REnaissance in Rural Europe: embedding diversity in organic and low-input food systems", this paper aimed to review recent research on the issue of health and nutritional cereal systems by adopting an innovative and participatory multi-actor approach which involved practitioners along with researchers. The participatory approach is the main innovation and peculiarity of this literature review. Nevertheless, the review highlights the many positive effects derived from eating whole and ancient grains such as a significant reduction in the risk of chronic diseases such as cancer, cardiovascular disease, and also a more favorable long-term weight management and increase in satiety. This review may be considered as a fruitful starting point that integrates research results to foster current and future healthier and sustainable practices in cereal systems