High glucose concentrations are required for endocrine pancreatic differentiation of mammalian adult fibroblasts.

Epigenetic conversion overcomes the stability of a terminally differentiated cell, allowing phenotypeswitch and providing an unlimited source of autologous cells of a different type. It is based on theexposure to an epigenetic modifier that increases cell plasticity, followed by a differentiation protocol.In our work we treat mammalian dermal fibroblasts with the demethylating agent 5-azacytidine. Celldifferentiation is directed toward the endocrine pancreatic lineage, with a sequential combination ofkey growth factors. The overall duration of the process is 36 days (Pennarossa, 2013; Brevini, 2015; Brevini,2015). However, this protocol, as well as all differentiation procedures described in the literature, useshigh and non-physiological concentrations of glucose. Here we report experiments aimed atinvestigating whether the use of lower glucose concentrations, that more closely mimic the in vivophysiological environment, can support fibroblast conversion into β-like cells. To do so, cells werecultured as described above, but using lower and more physiological glucose levels, namely 5.5 and 8.5mM that correspond to normoglycaemia before and after meals (International Diabetes Federation,2007). Our results show that mammalian cells are not able to differentiate into insulin secreting cells ina low glucose environment. In particular, cells do not aggregate into pancreatic islet structures anddisplay an altered gene expression pattern for several early pancreatic markers, when compared to thestandard trend obtained with 17.5 mM of glucose. These results suggest that high glucose levels areessential for the achievement of the endocrine pancreatic differentiation process in mammalian cellsand appear to be crucial for functional efficiency and morphological organization

Liquid Marble micro-bioreactor promotes 3D cell rearrangement and induces, maintains and stabilizes high plasticity in epigenetically erased fibroblasts

In the last years, many works demonstrated the possibility to directly interact with the epigenetic signature of an adult mature cell, through the use of epigenetic modifiers, (Pennarossa et al., 2013; Brevini et al., 2014, Chandrakantan et al., 2016) and new mechanisms underlying this process have been recently described (Manzoni et al., 2016). In particular, the small molecule 5-azacytidine (5-aza-CR) has been shown to induce a transient higher plasticity state in adult somatic cells, grown in standard 2D conditions. Recent evidence have also shown the possibility to regulate and maintain cell pluripotency through the use of 3D culture systems. In the experiments here presented, we combine the two approaches and investigate whether the simultaneous use of a 3D micro-bioreactor and 5-aza-CR is able to promote cell rearrangement, boost the induction of high plasticity and stably maintain it.To this purpose, fibroblasts were either plated on plastic dishes (2D) or encapsulated in a Liquid Marble (LM) micro-bioreactor (polytetrafluoroethylene (PTFE)), which has been previously shown to support the growth of living microorganisms, tumor spheroids, fibroblasts, red blood cells, and embryonic stem cells (Ledda et al., 2016). Cells were then erased with 5-aza-CR, for 18 hours and cultured in Embryonic Stem Cell (ESC) medium for up to 28 days. Morphological analysis and pluripotency related gene expression levels were monitored for the entire length of the experiments. 2D cells, kept a monolayer pattern and acquired a pluripotent state that was, however, transient and lost by day 6. In contrast the use of a 3D system maintained and stabilized the high plasticity state in LM cells until the end of the experiments (Fig. 1). The data obtained demonstrate that cell rearrangement and interactions may modulate 5-aza-CR induced plasticity and suggest a correlation between 3D mechano-transduction-related pathways and  epigenetic regulation of cell phenotype

Matrix stiffness boosts pancreatic differentiation via the YAP/TAZ mechanotransduction mediated pathway

In the last years, many papers highlighted the possibility to use epigenetic modifiers to directly interact with the epigenetic signature of an adult mature cell (Pennarossa et al., 2013; Chandrakantan et al., 2016). In particular, the molecule 5-azacytidine (5-aza-CR), which is able to interfere with DNA methylation, through both a direct and an indirect effect (Manzoni et al., 2016), can be used to remove the epigenetic ‘blocks’ responsible for tissue specification and to facilitate  cell transition to a different lineage. In parallel, recent evidence has also shown that epigenetic conversion is influenced by the 3D rearrangement and by the mechanical properties of the cellular microenvironment (Pennarossa et al., 2017). In the experiments here presented, we investigated the effect of a selected 3D culture system on the conversion process. We used INS-eGFP porcine fibroblasts, that express enhanced green fluorescent protein (eGFP) under the control of insulin gene promoter, as experimental model, and wild-type pig fibroblasts, as control. Both cell types, were plated either on plastic or on 1kPa polyacrylamide (PAA) gel, that mimics the stiffness of pancreatic tissue in vivo. Cells were erased with 5-aza-CR for 18h and exposed to specific differentiation stimuli for 36 days (Pennarossa et al., 2014). The use of INS-eGFP fibroblasts allowed real-time monitoring of cells progressing towards the pancreatic phenotype. Morphological analysis and pancreatic marker expression were checked for the entire length of the experiment. PAA gels encouraged the induction of islet-like structures, suggesting that the of tridimensional clusters may be a crucial aspect of pancreatic differentiation in vitro. Moreover, the use of an adequate substrate accelerated cell differentiation process and anticipated insulin secretion ability. The results obtained demonstrated the direct implication of the yes-associated protein/transcriptional co-activator with PDZ-binding motif (YAP/TAZ) mechanotransduction-mediated pathway, indicating  that mechanical cues exert a key role in pancreatic phenotype definition

Anatomical landmarks for tibial nerve motor branches in the management of spastic equinovarus foot after stroke: an ultrasonographic study

Objective: To identify the anatomical landmarks of tibial motor nerve branches to the gastrocnemii, soleus and tibialis posterior muscles for selective motor nerve blocks in the management of spastic equinovarus foot. Design: Observational study. Patients: Twenty-five chronic stroke patients with spastic equinovarus foot. Methods: Motor nerve branches to the gastrocnemii, soleus and tibialis posterior muscles were tracked in the affected leg, using ultrasonography, and located in the space (vertical, horizontal and deep) according to the position of the fibular head (proximal/distal) and a virtual line from the middle of the popliteal fossa to the Achilles tendon insertion (medial/lateral). Results: Mean coordinates for the gastrocnemius medialis motor branch were: 1.5 cm (standard deviation (SD) 2.7) vertical (proximal), 1.7 cm (SD 1.3) horizontal (medial), 1.1 cm (SD 0.4) deep; for the gastrocnemius lateralis motor branch: 0.9 cm (SD 2.2) vertical (proximal), 1.8 cm (SD 1.7) horizontal (lateral), 1.0 cm (SD 0.3) deep; for the soleus motor branch: 1.4 cm (SD 1.1) vertical (distal), 1.6 cm (SD 0.7) horizontal (lateral), 2.8 cm (SD 0.7) deep; and for the tibialis posterior motor branch: 4.3 cm (SD 1.5) vertical (distal), 1.9 cm (SD 0.9) horizontal (lateral), 4.2 cm (SD 0.8) deep. Conclusion: These findings may help in the identification of tibial motor nerve branches to the gastrocnemii, soleus and tibialis posterior muscles for selective motor nerve blocks in the management of spastic equinovarus foot

New HARPS and FEROS observations of GJ1046

In this paper we present new precise Doppler data of GJ1046 taken between November 2005 and July 2018 with the HARPS and the FEROS high-resolution spectographs. In addition, we provide a new stellar mass estimate of GJ1046 and we update the orbital parameters of the GJ1046 system. These new data and analysis could be used together with the GAIA epoch astrometry, when available, for braking the $\sin i$ degeneracy and revealing the true mass of the GJ1046 system.Comment: 2 pages, 1 figure, 1 table with RV data (available only in the Astro-PH version of the paper), Accepted by RNAA

Exploring emotional distress, psychological traits and attitudes in patients with chronic migraine undergoing OnabotulinumtoxinA prophylaxis versus withdrawal treatment

This explorative cross-sectional study aims at exploring emotional distress, psychological profiles, and the attitude towards receiving psychological support in eighty-seven patients with chronic migraine (CM) undergoing OnabotulinumtoxinA prophylactic treatment (OBT-A, n = 40) or withdrawal treatment (WT, n = 47). The outcomes were explored through a specific battery of questionnaires. 25% of patients undergoing OBT-A and almost half of the patients undergoing WT reported psychological distress of at least moderate-severe level, respectively. Coping strategies, self-efficacy, and perceived social support were similar in the two groups. Patients undergoing OBT-A presented lower psychological inflexibility than patients undergoing WT. Predictors of higher psychological distress were low perceived social support by friends, low self-efficacy, and higher avoidance strategies. In both groups, most of the patients evaluated receiving psychological support to be useful (79%). The potential beneficial effects of OBT-A on the severity of symptoms and psychological distress might further support its role in the multidisciplinary management of patients with CM. Identifying patients with psychological vulnerabilities who may benefit from psychological support is relevant in patients with CM

Screening for Mild Cognitive Impairment in Parkinson's Disease: Comparison of the Italian Versions of Three Neuropsychological Tests

Mild cognitive impairment (MCI) is frequent in Parkinson's disease (PD). Recently proposed criteria for MCI in PD (PD-MCI) indicate level I diagnosis based on abbreviated assessment and level II based on comprehensive neuropsychological evaluation. The study explored the sensitivity and specificity of the Italian versions of three neuropsychological tests for level I diagnosis of PD-MCI. We recruited 100 consecutive PD patients. After screening for inclusion criteria, 43 patients were included. The sensitivity and specificity of the Mini Mental State Examination (MMSE), the Montreal Cognitive Assessment (MoCA), and the Addenbrooke's Cognitive Examination Revised (ACE-R) in comparison to level II diagnosis of PD-MCI were examined. PD-MCI was diagnosed (level II) in 51% of patients. Disease duration was significantly longer and PD motor scales were more severely impaired in MCI group. The receiver-operator characteristics curve documented nonsignificant difference in the performance of the three tests, with slight advantage of MMSE (corrected data). The time of administration favored MMSE. In Italian-speaking PD patients, MMSE might represent a good screening tool for PD-MCI, because of the shorter time of administration and the performance comparable to those of MoCA and ACE-R. Further studies are needed to validate the new PD-MCI criteria across different languages and cultures

Improvement in motor symptoms, physical fatigue, and self-rated change perception in functional motor disorders: a prospective cohort study of a 12-week telemedicine program

Background: Functional motor disorders (FMDs) are highly disabling conditions associated with long-term disability, poor quality of life, and economic burden on health and social care. While multidisciplinary 5-days rehabilitation programs have been shown to reduce motor and non-motor symptoms, long-term management and monitoring in FMDs remain an unmet need. Aim: To compare a 12-weeks telemedicine program against a 12-weeks self-management program after a 5-days rehabilitation program for improving motor, non-motor symptoms, quality of life, and perception of change in patients with FMDs. Methods: The study population was 64 consecutive patients with a definite diagnosis of FMDs who underwent a 5-days in-person rehabilitation program followed by either a self-management (the first 32 patients) or a telemedicine program (the latter 32 patients). Validated measures of motor and non-motor symptoms such as fatigue and pain, quality of life, perception of change, gait, and postural control were recorded before (T0), after completion of rehabilitation (T1), and then again at 3 months (T2). Results: Improvement at 3-month follow-up assessment of motor symptoms (p < 0.001), physical fatigue (p = 0.028), and self-rated change perception (p = 0.043) was greater in the telemedicine group. No different between-groups effect was found on other dimensions of fatigue, pain, physical and mental health, and gait and postural control. Conclusions: Long-term management and expert monitoring of patients with FMDs via telemedicine may enhance long-term outcomes in motor symptoms and physical fatigue, with a positive long-term impact on self-rated health perception of change

Screening for Mild Cognitive Impairment in Parkinson’s Disease: Comparison of the Italian Versions of Three Neuropsychological Tests

Mild cognitive impairment (MCI) is frequent in Parkinson’s disease (PD). Recently proposed criteria for MCI in PD (PD-MCI) indicate level I diagnosis based on abbreviated assessment and level II based on comprehensive neuropsychological evaluation. The study explored the sensitivity and specificity of the Italian versions of three neuropsychological tests for level I diagnosis of PD-MCI. We recruited 100 consecutive PD patients. After screening for inclusion criteria, 43 patients were included. The sensitivity and specificity of the Mini Mental State Examination (MMSE), the Montreal Cognitive Assessment (MoCA), and the Addenbrooke’s Cognitive Examination Revised (ACE-R) in comparison to level II diagnosis of PD-MCI were examined. PD-MCI was diagnosed (level II) in 51% of patients. Disease duration was significantly longer and PD motor scales were more severely impaired in MCI group. The receiver-operator characteristics curve documented nonsignificant difference in the performance of the three tests, with slight advantage of MMSE (corrected data). The time of administration favored MMSE. In Italian-speaking PD patients, MMSE might represent a good screening tool for PD-MCI, because of the shorter time of administration and the performance comparable to those of MoCA and ACE-R. Further studies are needed to validate the new PD-MCI criteria across different languages and cultures

Comparison between physical and cognitive treatment in patients with MCI and Alzheimer's disease.

Cognitive and physical activity treatments (CT and PT) are two non-pharmacological approaches frequently used in patients with Mild Cognitive Impairment (MCI) and Alzheimer's Disease (AD). The aim of this study was to compare CT and PT in these diseases. Eighty-seven patients were randomly assigned to CT (n=30), PT (n=27) or control group (CTRL; n=30) for 6 months. The global cognitive function was measured by Mini Mental State Examination (MMSE). Specific neuropsychological tests explored attention, memory, executive functions, behavioral disorders. Cardiovascular risk factors (CVD) were collected. All measures were performed before (T0), after treatments (T1), and at three-months follow-up (T2). MMSE did not change from T0 to T1 and T2 in patients assigned to PT and CT, while CTRL patients showed a decline MCI: -11.8%, AD: -16.2%). Between group differences (MCI vs AD) were not found at T1 and T2. Significant worsening was found for CTRL in MCI (T0- T1: P=.039; T0-T2: P<.001) and AD (T0-T1: P<.001; T0-T2: P<.001), and amelioration was found for CT in AD (T0-T2: P<.001). Attention, executive functions and behavioral disorders were unaffected by either PT or CT. Memory was increased in patients with MCI assigned to PT (+6.9%) and CT (+8.5%).. CVD were ameliorated in the PT group. CTRL patients of both groups, revealed significant decline in all functions and no between groups differences were detected. PT appear to ameliorate CVD. Although between groups differences were not found, results suggest a major retention in MCI compared with AD, suggesting that the latter might benefit better of constant rather than periodic treatments. This study confirms the positive effects of CT and PT in mitigating the cognitive decline in MCI and AD patients, and it is the first to demonstrate their similar effectiveness on maintaining cognitive function