Collagenase isoforms for pancreas digestion.

The available information concerning the characteristics and composition of collagenase batches, which are effective in the digestion of human pancreas for islet transplants, is scarce and incomplete. A large inter-and intrabatched variability in activity and efficiency of blend enzymes available for isolation has been observed. The aim of this study was to characterize enzyme blend components. Liberase batches were characterized by SDS-PAGE analyses, microelectrophoresis, and then by MALDI-TOF MS analysis. Three main bands were detected by SDS-PAGE analysis and submitted to MALDI-TOF MS analysis. Two bands were found to correspond to class I (isoform β and another of 106 kDa) and one to class II (isoform δ) collagenase. These results represent an important step towards a complete characterization of enzymes, with the final aim of identifying key components for a standardized product

Predicting human cardiac QT alterations and pro-arrhythmic effects of compounds with a 3D beating heart-on-chip platform

Determining the potential cardiotoxicity and pro-arrhythmic effects of drug candidates remains one of the most relevant issues in the drug development pipeline. New methods enabling to perform more representative pre-clinical in vitro studies by exploiting induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) are under investigation to increase the translational power of the outcomes. Here we present a pharmacological campaign conducted to evaluate the drug-induced QT alterations and arrhythmic events on uHeart, a 3D miniaturized in-vitro model of human myocardium encompassing iPSC-CM and dermal fibroblasts embedded in fibrin. uHeart was mechanically trained resulting in synchronously beating cardiac microtissues in one week, characterized by a clear field potential (FP) signal that was recorded by means of an integrated electrical system. A drug screening protocol compliant with the new International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines was established and uHeart was employed for testing the effect of 11 compounds acting on single or multiple cardiac ion channels and well-known to elicit QT prolongation or arrhythmic events in clinics. The alterations of uHeart's electrophysiological parameters such as the beating period, the FP duration, the FP amplitude and the detection of arrhythmic events prior and after drug administration at incremental doses were effectively analyzed through a custom developed algorithm. Results demonstrated the ability of uHeart to successfully anticipate clinical outcome and to predict the QT prolongation with a sensitivity of 83.3%, a specificity of 100% and an accuracy of 91.6%. Cardiotoxic concentrations of drugs were notably detected in the range of the clinical highest blood drug concentration (Cmax), qualifying uHeart as a fit-to-purpose pre-clinical tool for cardiotoxicity studies

The Alto Tiberina Near Fault Observatory (northern Apennines, Italy)

The availability of multidisciplinary and high-resolution data is a fundamental requirement to understand the physics of earthquakes and faulting. We present the Alto Tiberina Near Fault Observatory (TABOO), a research infrastructure devoted to studying preparatory processes, slow and fast deformation along a fault system located in the upper Tiber Valley (northern Apennines), dominated by a 60 km long low-angle normal fault (Alto Tiberina, ATF) active since the Quaternary. TABOO consists of 50 permanent seismic stations covering an area of 120 × 120 km2. The surface seismic stations are equipped with 3-components seismometers, one third of them hosting accelerometers. We instrumented three shallow (250 m) boreholes with seismometers, creating a 3-dimensional antenna for studying micro-earthquakes sources (detection threshold is ML 0.5) and detecting transient signals. 24 of these sites are equipped with continuous geodetic GPS, forming two transects across the fault system. Geochemical and electromagnetic stations have been also deployed in the study area. In 36 months TABOO recorded 19,422 events with ML ≤ 3.8 corresponding to 23.36e-04 events per day per squared kilometres; one of the highest seismicity rate value observed in Italy. Seismicity distribution images the geometry of the ATF and its antithetic/synthetic structures located in the hanging-wall. TABOO can allow us to understand the seismogenic potential of the ATF and therefore contribute to the seismic hazard assessment of the area. The collected information on the geometry and deformation style of the fault will be used to elaborate ground shaking scenarios adopting diverse slip distributions and rupture directivity models.PublishedS03275T. Sismologia, geofisica e geologia per l'ingegneria sismicaJCR Journa

Allogeneic islet transplantation

Significant progress has been made in the field of β-cell replacement therapies by islet transplantation in patients with unstable Type 1 diabetes mellitus (T1DM). Recent clinical trials have shown that islet transplantation can reproducibly lead to insulin independence when adequate islet numbers are implanted. Benefits include improvement of glycemic control, prevention of severe hypoglycemia and amelioration of quality of life. Numerous challenges still limit this therapeutic option from becoming the treatment of choice for T1DM. The limitations are primarily associated with the low islet yield of human pancreas isolations and the need for chronic immunosuppressive therapies. Herein the authors present an overview of the historical progress of islet transplantation and outline the recent advances of the field. Cellular therapies offer the potential for a cure for patients with T1DM. The progress in β-cell replacement treatment by islet transplantation as well as those of emerging immune interventions for the restoration of self tolerance justify great optimism for years to come

Cancer-on-chip: a 3D model for the study of the tumor microenvironment

Abstract The approval of anticancer therapeutic strategies is still slowed down by the lack of models able to faithfully reproduce in vivo cancer physiology. On one hand, the conventional in vitro models fail to recapitulate the organ and tissue structures, the fluid flows, and the mechanical stimuli characterizing the human body compartments. On the other hand, in vivo animal models cannot reproduce the typical human tumor microenvironment, essential to study cancer behavior and progression. This study reviews the cancer-on-chips as one of the most promising tools to model and investigate the tumor microenvironment and metastasis. We also described how cancer-on-chip devices have been developed and implemented to study the most common primary cancers and their metastatic sites. Pros and cons of this technology are then discussed highlighting the future challenges to close the gap between the pre-clinical and clinical studies and accelerate the approval of new anticancer therapies in humans

A catalogue of non-tectonic earthquakes in central-eastern Italy

Quarry explosions and other non-tectonic signals can contaminate seismic catalogues, especially in areas where dense seismic networks allow to detect even low energy events. This paper presents the algorithm adopted for the discrimination of non-tectonic earthquakes in central-eastern Italy, an area with a high rate of events of this kind (4500 were recorded in the 1996-2012 period). Starting from an empirical classification based on the analysis of areas in which the ratio between daytime and nighttime events is strongly anomalous, a waveform similarity approach allows to simplify the procedure of detection making the final classification more robust. The resulting catalogue of non-tectonic earthquakes (quarry blasts and other anomalous signals) is a useful tool for anyone wanting to carry out a careful analysis of the tectonic seismicity of central-eastern Italy.PublishedS03284T. Sismologia, geofisica e geologia per l'ingegneria sismicaJCR Journa

Automated control procedures and first results from the temporary seismic monitoring of the 2012 Emilia sequence

After moderate to strong earthquakes in Italy or in the surrounding areas, the Istituto Nazionale di Geofisica e Vulcanologia (INGV; National Institute for Geophysics and Volcanology) activates a temporary seismic network infrastructure. This is devoted to integration with the Italian National Seismic Network (RSN) [Delladio 2011] in the epicentral area, thus improving the localization of the aftershocks distribution after a mainshock. This infrastructure is composed of a stand-alone, locally recording part (Re.Mo.) [Moretti et al. 2010] and a real-time telemetered part (Re.Mo.Tel.) [Abruzzese et al. 2011a, 2011b] that can stream data to the acquisition centers in Rome and Grottaminarda. After the May 20, 2012, Ml 5.9 earthquake in the Emilia region (northern Italy), the temporary network was deployed in the epicentral area; in particular, 10 telemetered and 12 stand-alone stations were installed [Moretti et al. 2012, this volume]. Using the dedicated connection between the acquisition center in Rome and the Ancona acquisition sub-center [Cattaneo et al. 2011], the signals of the real-time telemetered stations were acquired also in this sub-center. These were used for preliminary quality control, by adopting the standard procedures in use here (see next paragraph, and Monachesi et al. [2011]). The main purpose of the present study is a first report on this quality check, which should be taken into account for the correct use of these data. […

The “ON-OFF” Switching Response of Reactive Oxygen Species in Acute Normobaric Hypoxia: Preliminary Outcome

Exposure to acute normobaric hypoxia (NH) elicits reactive oxygen species (ROS) accumulation, whose production kinetics and oxidative damage were here investigated. Nine subjects were monitored while breathing an NH mixture (0.125 FIO2 in air, about 4100 m) and during recovery with room air. ROS production was assessed by Electron Paramagnetic Resonance in capillary blood. Total antioxidant capacity, lipid peroxidation (TBARS and 8-iso-PFG2α), protein oxidation (PC) and DNA oxidation (8-OH-dG) were measured in plasma and/or urine. The ROS production rate (μmol·min−1) was monitored (5, 15, 30, 60, 120, 240 and 300 min). A production peak (+50%) was reached at 4 h. The on-transient kinetics, exponentially fitted (t1/2 = 30 min r2 = 0.995), were ascribable to the low O2 tension transition and the mirror-like related SpO2 decrease: 15 min: −12%; 60 min: −18%. The exposure did not seem to affect the prooxidant/antioxidant balance. Significant increases in PC (+88%) and 8-OH-dG (+67%) at 4 h in TBARS (+33%) one hour after hypoxia offset were also observed. General malaise was described by most of the subjects. Under acute NH, ROS production and oxidative damage resulted in time and SpO2-dependent reversible phenomena. The experimental model could be suitable for evaluating the acclimatation level, a key element in the context of mountain rescues in relation to technical/medical workers who have not had enough time for acclimatization—as, for example, during helicopter flights

Effects of Combined Physical and Cognitive Virtual Reality-Based Training on Cognitive Impairment and Oxidative Stress in MCI Patients: A Pilot Study

none12noThe growing elderly population and the increased incidence of mild cognitive impairment (MCI) and Alzheimer's disease (AD) call for the improvement of the quality and the efficacy of the healthcare and social support services. Exercise and cognitive stimulation have been demonstrated to mitigate cognitive impairment and oxidative stress (OxS) has been recognized as a factor that contributes to the advancement of neurodegenerative diseases. Taking these aspects into account, the impact of a novel virtual reality (VR)-based program combining aerobic exercise and cognitive training has been evaluated in the pilot study proposed here. Ten patients (aged 73.3 ± 5.7 years) with MCI (Mini-Mental State Examination, MMSE: 23.0 ± 3.4) were randomly assigned to either 6 weeks physical and cognitive training (EXP) or control (CTR) group. Evaluations of cognitive profile, by a neuropsychological tests battery, and OxS, by collection of blood and urine samples, were performed before and at the end of the experimental period. The assessment of the patients' opinions toward the intervention was investigated through questionnaires. EXP group showed a tendency towards improvements in the MMSE, in visual-constructive test and visuo-spatial tests of attention, while CTR worsened. EXP group showed a greater improvement than CTR in the executive test, memory functions and verbal fluency. No statistical significance was obtained when comparing within and between both the groups, probably due to small number of subjects examined, which amplifies the effect of the slight heterogeneity in scores recorded. Despite a greater worsening of Daily Living Activities tests, all participants reported a better performance in real life, thanks to the elicited self-perceived improvement. After training intervention OxS (i.e., reactive oxygen species (ROS) production, oxidative damage of lipids and DNA) decreased resulting in significantly (range p < 0.05-0.001) lower in EXP vs. CTR group. Although not conclusive, the recorded effects in the present study are promising and suggest that this proposal would be a useful tool in support of cognitive training reducing OxS too. However, further studies on larger scale samples of patients are needed.noneMrakic-Sposta, Simona*; Di Santo, Simona G.; Franchini, Flaminia; Arlati, Sara; Zangiacomi, Andrea; Greci, Luca; Moretti, Sarah; Jesuthasan, Nithiya; Marzorati, Mauro; Rizzo, Giovanna; Sacco, Marco; Vezzoli, AlessandraMrakic-Sposta, Simona; Di Santo, Simona G.; Franchini, Flaminia; Arlati, Sara; Zangiacomi, Andrea; Greci, Luca; Moretti, Sarah; Jesuthasan, Nithiya; Marzorati, Mauro; Rizzo, Giovanna; Sacco, Marco; Vezzoli, Alessandr

Performance of earthquake early warning systems during the 2016-2017 Mw5-6.5 central Italy sequence

Earthquake early warning systems (EEWSs) are nowadays contributing to seismic risk mitigation actions, both in terms of losses and societal resilience, by issuing an alert promptly after the earthquake origin and before the ground-shaking impacts the target to be protected. In this work, we analyze the performance of network-based and stand-alone (on-site) early warning systems during the 2016–2017 central Italy sequence, characterized by events with magnitude as large as 6.5. For the largest magnitude event, both systems predict well the ground shaking nearby the event source, with a rate of success in the 85%–90% range, within the potential earthquake damage zone. However, the lead time, that is, the time available for security actions, is significantly larger for the network-based system. For the regional system, it increases to more than 10 s at 40 km from the event epicenter. The stand-alone system performs better in the near-source region, still showing a positive albeit small lead time (60 km), the performances slightly degrade, mostly owing to the large uncertainty in the attenuation relationships. This study opens up the possibility for making an operational EEWS in Italy, based on the available acceleration networks, provided that the delay due to data telemetry has to be reduced.Published1-125T. Sismologia, geofisica e geologia per l'ingegneria sismicaJCR Journa