Porcine Small Intestinal Submucosa (SIS) as a Suitable Scaffold for the Creation of a Tissue-Engineered Urinary Conduit: Decellularization,Biomechanical and Biocompatibility Characterization Using New Approaches

Bladder cancer (BC) is among the most common malignancies in the world and a relevant cause of cancer mortality. BC is one of the most frequent causes for bladder removal through radical cystectomy, the gold-standard treatment for localized muscle-invasive and some cases of high-risk, non-muscle-invasive bladder cancer. In order to restore urinary functionality, an autologous intestinal segment has to be used to create a urinary diversion. However, several complications are associated with bowel-tract removal, affecting patients’ quality of life. The present study project aims to develop a bio-engineered material to simplify this surgical procedure, avoiding related surgical complications and improving patients’ quality of life. The main novelty of such a therapeutic approach is the decellularization of a porcine small intestinal submucosa (SIS) conduit to replace the autologous intestinal segment currently used as urinary diversion after radical cystectomy, while avoiding an immune rejection. Here, we performed a preliminary evaluation of this acellular product by developing a novel decellularization process based on an environmentally friendly, mild detergent, i.e., Tergitol, to replace the recently declared toxic Triton X-100. Treatment efficacy was evaluated through histology, DNA, hydroxyproline and elastin quantification, mechanical and insufflation tests, two-photon microscopy, FTIR analysis, and cytocompatibility tests. The optimized decellularization protocol is effective in removing cells, including DNA content, from the porcine SIS, while preserving the integrity of the extracellular matrix despite an increase in stiffness. An effective sterilization protocol was found, and cytocompatibility of treated SIS was demonstrated from day 1 to day 7, during which human fibroblasts were able to increase in number and strongly organize along tissue fibres. Taken together, this in vitro study suggests that SIS is a suitable candidate for use in urinary diversions in place of autologous intestinal segments, considering the optimal results of decellularization and cell proliferation. Further efforts should be undertaken in order to improve SIS conduit patency and impermeability to realize a future viable substitute.     

Pediatric Brain Tumors: Signatures from the Intact Proteome

The present investigation aimed to explore the intact proteome of tissues of pediatric brain tumors of different WHO grades and localizations, including medulloblastoma, pilocytic astrocytoma, and glioblastoma, in comparison with the available data on ependymoma, to contribute to the understanding of the molecular mechanisms underlying the onset and progression of these pathologies. Tissues have been homogenized in acidic water–acetonitrile solutions containing proteases inhibitors and analyzed by LC–high resolution MS for proteomic characterization and label-free relative quantitation. Tandem MS spectra have been analyzed by either manual inspection or software elaboration, followed by experimental/theoretical MS fragmentation data comparison by bioinformatic tools. Statistically significant differences in protein/peptide levels between the different tumor histotypes have been evaluated by ANOVA test and Tukey’s post-hoc test, considering a p-value > 0.05 as significant. Together with intact protein and peptide chains, in the range of molecular mass of 1.3–22.8 kDa, several naturally occurring fragments from major proteins, peptides, and proteoforms have been also identified, some exhibiting proper biological activities. Protein and peptide sequencing allowed for the identification of different post-translational modifications, with acetylations, oxidations, citrullinations, deamidations, and C-terminal truncations being the most frequently characterized. C-terminal truncations, lacking from two to four amino acid residues, particularly characterizing the β-thymosin peptides and ubiquitin, showed a different modulation in the diverse tumors studied. With respect to the other tumors, medulloblastoma, the most frequent malignant brain tumor of the pediatric age, was characterized by higher levels of thymosin β4 and β10 peptides, the latter and its des-IS form particularly marking this histotype. The distribution pattern of the C-terminal truncated forms was also different in glioblastoma, particularly underlying gender differences, according to the definition of male and female glioblastoma as biologically distinct diseases. Glioblastoma was also distinguished for the peculiar identification of the truncated form of the α-hemoglobin chain, lacking the C-terminal arginine, and exhibiting oxygen-binding and vasoconstrictive properties different from the intact form. The proteomic characterization of the undigested proteome, following the top-down approach, was challenging to originally investigate the post-translational events that differently characterize pediatric brain tumors. This study provides a contribution to elucidate the molecular profiles of the solid tumors most frequently affecting the pediatric age, and which are characterized by different grades of aggressiveness and localization.     

Host Restriction Factors Modulating HIV Latency and Replication in Macrophages

In addition to CD4⁺ T lymphocytes, myeloid cells and, particularly, differentiated macrophages are targets of human immunodeficiency virus type-1 (HIV-1) infection via the interaction of gp120Env with CD4 and CCR5 or CXCR4. Both T cells and macrophages support virus replication, although with substantial differences. In contrast to activated CD4⁺ T lymphocytes, HIV-1 replication in macrophages occurs in nondividing cells and it is characterized by the virtual absence of cytopathicity both in vitro and in vivo. These general features should be considered in evaluating the role of cell-associated restriction factors aiming at preventing or curtailing virus replication in macrophages and T cells, particularly in the context of designing strategies to tackle the viral reservoir in infected individuals receiving combination antiretroviral therapy. In this regard, we will here also discuss a model of reversible HIV-1 latency in primary human macrophages and the role of host factors determining the restriction or reactivation of virus replication in these cells.     

An automatic procedure for the synthesis and optimization of very low-frequency Gm–C integrators

Fully integrated low frequency filters are critical cells that should be carefully designed in order to avoid excessive area occupation. In this work we propose an automatic procedure capable of optimizing the design of G_m?CC integrators, which constitute the basis of a wide class of G_m?CC filters. The optimization target is minimizing the cell area with constraints on input range and low frequency noise. Lower and upper bounds can be fixed to most quantities and design parameters in order to avoid solutions that are not compatible with the physical limitations of the process. The program has been developed within the MATLAB^? platform, exploiting the optimization toolbox. The effect of several important design parameters on the optimization of low frequency integrators has been investigated using the proposed routine. The strong interaction between noise and low frequency constraints has been demonstrated, showing the impressive impact of strict noise specifications on the occupied area. The actual effectiveness of parameters such as the current division factor or approaches such as flicker noise rejection by means of chopper modulation has been investigated. Examples of integrator synthesis, performed using the proposed procedure configured with the parameters of a commercial CMOS process, are presented. The consistence between the characteristics of the cells and the initial specifications has been checked using electrical simulations showing a maximum discrepancy with the initial specifications of nearly 80%. A semi-manual method to refine the synthesized cells and improve the accuracy is proposed.     

Low computational cost CFD analysis of thermoacoustic oscillations

The numerical analysis of thermoacoustic oscillation phenomena by means of time-dependent CFD simulations usually requires a great computational effort, which may not be reasonable in industrial design. On the other hand, CFD tools provide the only approach that includes all the physical and chemical aspects involved in the thermoacoustic coupling between flame heat release and the acoustic modes of the burner/combustion chamber system. This paper presents some guidelines to reduce the computational effort required to perform a CFD analysis of the thermoacoustic oscillations with commercial codes. These guidelines are organized in a procedure that can be followed to analyze thermoacoustic coupling conditions that actually lead to unstable oscillations or are identified as potentially critical in the design phase. This procedure is also illustrated by an example of application, the partially-premixed flame type burner of a real 10 MW industrial boiler which shows noisy pressure fluctuations at a low frequency.     

Disaster-survivable cloud-network mapping

Cloud-computing services are provided to consumers through a network of servers and network equipment. Cloud-network (CN) providers virtualize resources [e.g., virtual machine (VM) and virtual network (VN)] for efficient and secure resource allocation. Disasters are one of the worst threats for CNs as they can cause massive disruptions and CN disconnection. A disaster may also induce post-disaster correlated, cascading failures which can disconnect more CNs. Survivable virtual-network embedding (SVNE) approaches have been studied to protect VNs against single physical-link/-node and dual physical-link failures in communication infrastructure, but massive disruptions due to a disaster and their consequences can make SVNE approaches insufficient to guarantee cloud-computing survivability. In this work, we study the problem of survivable CN mapping from disaster. We consider risk assessment, VM backup location, and post-disaster survivability to reduce the risk of failure and probability of CN disconnection and the penalty paid by operators due to loss of capacity. We formulate the proposed approach as an integer linear program and study two scenarios: a natural disaster, e.g., earthquake and a human-made disaster, e.g., weapons-of-mass-destruction attack. Our illustrative examples show that our approach reduces the risk of CN disconnection and penalty up to 90 % compared with a baseline CN mapping approach and increases the CN survivability up to 100 % in both scenarios.     

Radiant floors integrated with PCM for indoor temperature control

The control of indoor thermal comfort in buildings through thermal inertia during the summer season plays a fundamental role in the design of energy efficient buildings, especially in the Mediterranean climate. In fact, lightweight, highly insulated buildings cannot provide the necessary mass to buffer thermal gains. Phase change materials (PCM) have been used to provide lightweight building components with the required thermal inertia without increasing their overall mass. So far the integration of PCM into lightweight piped radiant floors for the control of thermal comfort during summer cooling regimes has not been investigated. This paper reports the development of a lightweight piped radiant floor prototype with an integrated PCM layer aimed at buffering internal gains at constant temperature during summer cooling regimes without affecting its winter warming capacity. Both the construction of the laboratory specimens and the development of the optimized finite element models are detailed and the assessment of the floor performance in a simulated room is discussed.     

Prediction of point-defect migration energy barriers in alloys using artificial intelligence for atomistic kinetic Monte Carlo applications

We significantly improved a previously proposed method to take into account chemical and also relaxation effects on point-defect migration energy barriers, as predicted by an interatomic potential, in a rigid lattice atomistic kinetic Monte Carlo simulation. Examples of energy barriers are rigorously calculated, including chemical and relaxation effects, as functions of the local atomic configuration, using a nudged elastic bands technique. These examples are then used to train an artificial neural network that provides the barriers on-demand during the simulation for each configuration encountered by the migrating defect. Thanks to a newly developed training method, the configuration can include a large number of neighbour shells, thereby properly including also strain effects. Satisfactory results have been obtained when the configuration includes different chemical species only. The problems encountered in the extension of the method to configurations including any number of point-defects are stated and solutions to tackle them are sketched.