Gut acellular matrix for the in vitro study of Enteric Nervous System cells

Enteric nervous system (ENS) cells respond to the intestinal extracellular matrix (ECM) signals changing their proliferation rate, migration and differentiation. In this study, we explored in vitro the adaptive response of primary ENS cell cultures to the stimulation of gut acellular matrix (AM) defining the gene expression profile of neuronal functionality markers. Scanning electron microscopy was used to detect the acquisition of specific morphological features. Intestinal AM was prepared using an enzyme-detergent treatment. Primary rat enteric cells were isolated from the myenteric plexus of postnatal rats using an enzymatic method and seeded on intestinal AM in the presence of exogenous neurotrophic factors. The morphological properties and the expression of specific differentiation markers were evaluated by Scanning Electron Microscopy (SEM) and wholemount fluorescent staining. In order to verify the synergic activity of soluble factors and AM, the gene expression of neurotransmitter receptors was evaluated by qPCR in ENS cells cultured in SM conditions in the presence or not of AM. The development of interconnected ganglion-like structures and the expression of neurotransmitter receptors suggested that gut matrix engineered with ENS cells could be useful for medical applications of regenerative medicine or for the in vitro assessment of tridimensional culture system of ENS

Discourse-centric learning analytics

Drawing on sociocultural discourse analysis and argumentation theory, we motivate a focus on learners' discourse as a promising site for identifying patterns of activity which correspond to meaningful learning and knowledge construction. However, software platforms must gain access to qualitative information about the rhetorical dimensions to discourse contributions to enable such analytics. This is difficult to extract from naturally occurring text, but the emergence of more-structured annotation and deliberation platforms for learning makes such information available. Using the Cohere web application as a research vehicle, we present examples of analytics at the level of individual learners and groups, showing conceptual and social network patterns, which we propose as indicators of meaningful learning

A Model of Porous Catalyst Accounting for Incipiently Non-isothermal Effects*

An approximate model accounting for incipiently non-isothermal effects is derived from a well-known model of porous catalyst for appropriate, realistic limiting values of the parameters. In this limit, the original model is a singularly perturbed, m-D reaction–diffusion system, and the approximate model is given by the m-D heat equation with nonlinear boundary condition, coupled with infinitely many (ifm2) 1-D semilinear parabolic equations, one for each point of the boundary of the spatial domain. Some limiting cases are still considered in the approximate model that lead to further simplifications

A service-learning experience in a free medical centre for undocumented migrants and homeless people

Background: Service-learning experiences, informed by the realities of poverty and marginalization, are important for the education of future health professionals in order to commit them to tackling health inequalities and working with underserved populations. At the Caritas Medical Centre for undocumented migrants and homeless in Rome, students obtain an educational experience of service. The aim of this study is to try to measure the long-term impact of this experience on the professional and life choices of the student participants. Methods: A questionnaire was designed and distributed by email to all 19–29 years old participants in the experience. Responses were collected and analysed in a quantitative descriptive way and in a qualitative way using the knowledge, skills and attitudes model. Results: One hundred and seven students responded from the total 763 questionnaires distributed. Ninety-five percent of participants expressed a very high overall satisfaction, 93% declared that the experience influenced his/her future personal choices, and 84% found that the experience influenced their professional choices. Results were arranged into 6 categories of comments: knowledge about the realities of migration, poverty, and marginalization; relational skills; collaborative skills; attitudes towards migrants, poor people and others; Attitudes towards future professions; Attitudes towards life. A final category was listed with self-reflective questions related to the experience. Conclusion: This research shows the importance of service-learning experiences made during academic studies from young students of medicine and other faculties. Developing a relationship with marginalized and homeless people, within a voluntary service setting, can influence the future professional and personal choices of students. Universities should recognize the value of such experiences and establish partnerships with non-profit organizations to allow future health professionals to confront health inequities and commit themselves to their reduction

Growth and Differentiation of Circulating Stem Cells After Extensive Ex Vivo Expansion

Background:: Stem cell therapy is gaining momentum as an effective treatment strategy for degenerative diseases. Adult stem cells isolated from various sources (i.e., cord blood, bone marrow, adipose tissue) are being considered as a realistic option due to their well-documented therapeutic potentials. Our previous studies standardized a method to isolate circulating multipotent cells (CMCs) that are able to sustain long term in vitro culture and differentiate towards mesodermal lineages. Methods:: In this work, long-term cultures of CMCs were stimulated to study in vitro neuronal and myogenic differentiation. After induction, cells were analysed at different time points. Morphological studies were performed by scanning electron microscopy and specific neuronal and myogenic marker expression were evaluated using RT-PCR, flow cytometry and western blot. For myogenic plasticity study, CMCs were transplanted into in vivo model of chemically-induced muscle damage. Results:: After neurogenic induction, CMCs showed characteristic dendrite-like morphology and expressed specific neuronal markers both at mRNA and protein level. The calcium flux activity of CMCs under stimulation with potassium chloride and the secretion of noradrenalin confirmed their ability to acquire a functional phenotype. In parallel, the myogenic potential of CMCs was confirmed by their ability to form syncytium-like structures in vitro and express myogenic markers both at early and late phases of differentiation. Interestingly, in a rat model of bupivacaine-induced muscle damage, CMCs integrated within the host tissue taking part in tissue repair. Conclusion:: Overall, collected data demonstrated long-term cultured CMCs retain proliferative and differentiative potentials suggesting to be a good candidate for cell therapy

The Physical activity behaviour of vulnerable diabetic migrants: the role of juridical status

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Infrapatellar Fat Pad Stem Cells Responsiveness to Microenvironment in Osteoarthritis: From Morphology to Function

Recently, infrapatellar fat pad (IFP) has been considered as a source of stem cells for cartilage regeneration in osteoarthritis (OA) due to their ability for differentiation into chondrocytes. However, stressful conditions, like that related to OA, may induce a pathogenic reprograming. The aim of this study was to characterize the structural and functional properties of a new population of stem cells isolated from osteoarthritic infrapatellar fat pad (OA-IFP). Nine OA patients undergoing total knee arthroplasty (TKA) were enrolled in this study [median age = 74 years, interquartile range (IQR) = 78.25-67.7; median body mass index = 29.4 Kg/m2, IQR = 31.7-27.4]. OA-IFP stem cells were isolated and characterized for morphology, stemness, metabolic profile and multi-differentiative potential by transmission electron microscopy, flow cytometric analysis, gene expression study and cytochemistry. OA-IFP stem cells displayed a spindle-like morphology, self-renewal potential and responsiveness (CD44, CD105, VEGFR2, FGFR2, IL1R, and IL6R) to microenvironmental stimuli. Characterized by high grade of stemness (STAT3, NOTCH1, c-Myc, OCT-4, KLF4, and NANOG), the cells showed peculiar immunophenotypic properties (CD73+/CD39+/CD90+/CD105+/CD44\u2013/+/CD45\u2013). The expression of HLA-DR, CD34, Fas and FasL was indicative of a possible phenotypic reprograming induced by inflammation. Moreover, the response to mechanical stimuli together with high expression level of COL1A1 gene, suggested their possible protective response against in vivo mechanical overloading. Conversely, the low expression of CD38/NADase was indicative of their inability to counteract NAD+-mediated OA inflammation. Based on the ultrastructural, immunophenotypic and functional characterization, OA-IFP stem cells were hypothesized to be primed by the pathological environment and to exert incomplete protective activity from OA inflammation

Inflammatory profile of neurotrophins, IL-6, IL1-β, TNF-α, VEGF, ICAM-1 and TGF-β in the Human Waldeyer’s ring

The palatine tonsils, nasopharyngeal tonsil (adenoid) and lingual tonsil constitute the major part of Waldeyer’s ring, with the tubal tonsils and lateral pharyngeal bands as less prominent components. The lymphoid tissue of Waldeyer’s ring is located at the gateway of the respiratory and alimentary tract and belongs to the mucosa-associated lymphoid tissue (MALT). The lymphatic tissue is known to interact with the nervous system and several organs implicated in the host response to a wide range of stressors (Otten et al., 1995; Kaneko et al., 2012; Ogasawara et al., 2011). This study focusses on the expression of some neurotrophins (NTs), their high- and low-affinity receptors in human adenoid tissues, lingual and palatine tonsils via immunohistochemical analysis, as well as on the expression of some inflammatory cytokines and other tissue growth factors (IL-6, IL1-β, TNF-α, VEGF, ICAM-1 and TGF-β)). Light microscopy immunohistochemistry performed on human samples showed to be generally positive for all the NTs investigated (NGF, BDNF, NT-3) and their receptors (TrKA, TrKB and TrKC) as well as the other cytokines and growth factors studied with some different expression levels. Real time PCR analysis is in progress to quantitate these data. Our data corroborate previous studies, suggesting that neurotrophins and inflammatory cytokines may mediate functional signals in lymphoid aggregates (Yusuf-Makagiansar et al., 2002; Ruoco et al., 1990)

S100B protein as a therapeutic target in multiple sclerosis: The S100B inhibitor arundic acid protects from chronic experimental autoimmune encephalomyelitis

S100B is an astrocytic protein behaving at high concentration as a damage-associated molecular pattern molecule. A direct correlation between the increased amount of S100B and inflammatory processes has been demonstrated, and in particular, the inhibitor of S100B activity pentamidine has been shown to ameliorate clinical scores and neuropathologic-biomolecular parameters in the relapsing-remitting experimental autoimmune encephalomyelitis mouse model of multiple sclerosis. This study investigates the effect of arundic acid (AA), a known inhibitor of astrocytic S100B synthesis, in the chronic experimental autoimmune encephalomyelitis, which is another mouse model of multiple sclerosis usually studied. By the daily evaluation of clinical scores and neuropathologic-molecular analysis performed in the spinal cord, we observed that the AA-treated group showed lower severity compared to the vehicle-treated mice, particularly in the early phase of disease onset. We also observed a significant reduction of astrocytosis, demyelination, immune infiltrates, proinflammatory cytokines expression and enzymatic oxidative reactivity in the AA-treated group. Overall, our results reinforce the involvement of S100B in the development of animal models of multiple sclerosis and propose AA targeting the S100B protein as a focused potential drug to be considered for multiple sclerosis treatment