Integratable non-clogging microconcentrator based on counter-flow principle for continuous enrichment of CaSki cells sample

This study addresses the need to reduce the risk of clogging when preparing samples for cell concentration, i.e., the CaSki Cell-lines (epidermoid cervical carcinoma cells). Aiming to develop a non-clogging microconcentrator, we proposed a new counter-flow concentration unit characterized by the directions of penetrating flows being at an obtuse angle to the main flow, due to employment of streamlined turbine blade-like micropillars. Based on the optimization results of the counter-flow unit profile, a fractal arrangement for the counter-flow concentration unit was developed. A counter-flow microconcentrator chip was then designed and fabricated, with both the processing layer and collecting layer arranged in terms of the honeycomb structure. Visualized experiments using CaSki cell samples on the microconcentrator chip demonstrated that no cell-clogging phenomena occurred during the test and that no cells were found in the final filtrate. The test results show an excellent concentration performance for the microconcentrator chip, while a concentrating ratio of >4 with the flow rate being below 1.0 ml/min. As only geometrical structure is employed in the passive device, the counter-flow microconcentrator can be easily integrated into advanced microfluidic systems. Owing to the merit of non-clogging and continuous processing ability, the counter-flow microconcentrator is not only suitable for the sample preparation within biomedical field, but also applicable in water-particle separation.     

Computational study on the interaction of a ring-hydroxylating dioxygenase from Sphingomonas CHY-1 with PAHs

The massive computational resources available in the framework of a grid paradigm approach represent an emerging tool in the bioinformatics field. In this paper, we used the above approach in the rapid determination of the interactions between the ring-hydroxylating dioxygenase, comprised six enzymatic subunits, and polycyclic aromatic hydrocarbons (PAHs) in their optimal positions. The results were obtained by simulating enzyme dynamics at 300 K through molecular dynamics calculations. For the first time, the equilibrated structure of the dioxygenase revealed a network of channels throughout the enzyme that were sufficiently large to allow a flow of small ions or molecules from the inner core of the complex to its exterior surface. The ring-hydroxylating dioxygenase was able to interact with some of the studied PAHs. Additionally, not only the number of aromatic rings but also the PAH shape were critical in predicting the ability of the dioxygenase to interact with these types of molecules. Docking calculations shed light on a new possible binding site that is far from the enzymatic one, which is potentially interesting in considering the stability of the enzyme itself.     

Active fault tolerant control of discrete event systems using online diagnostics

The aim of this paper is to deal with the problem of fault tolerant control in the framework of discrete event systems modeled as automata. A fault tolerant controller is a controller able to satisfy control specifications both in nominal operation and after the occurrence of a fault. This task is solved by means of a parameterized controller that is suitably updated on the basis of the information provided by online diagnostics: the supervisor actively reacts to the detection of a malfunctioning component in order to eventually meet degraded control specifications. Starting from an appropriate model of the system, we recall the notion of safe diagnosability as a necessary step in order to achieve fault tolerant control. We then introduce two new notions: (i) “safe controllability”, which represents the capability, after the occurrence of a fault, of steering the system away from forbidden zones and (ii) “active fault tolerant system”, which is the property of safely continuing operation after faults. Finally, we show how the problem can be solved using a general control architecture based on the use of special kind of diagnoser, called “diagnosing controller”, which is used to safely detect faults and to switch between the nominal control policy and a bank of reconfigured control policies. A simple example is used to illustrate the new notions and the control architecture introduced in the paper.     

Experiments on the reliability of stochastic spiking neural P systems

In the area of membrane computing, time-freeness has been defined as the ability for a timed membrane system to produce always the same result, independently of the execution times associated to the rules. In this paper, we use a similar idea in the framework of spiking neural P systems, a model inspired by the structure and the functioning of neural cells. In particular, we introduce stochastic spiking neural P systems where the time of firing for an enabled spiking rule is probabilistically chosen and we investigate when, and how, these probabilities can influence the ability of the systems to simulate, in a reliable way, universal machines, such as register machines.     

Adaptive course generation through learning styles representation

This paper presents an approach to automatic course generation and student modeling. The method has been developed during the European funded projects Diogene and Intraserv, focused on the construction of an adaptive e-learning platform. The aim of the platform is the automatic generation and personalization of courses, taking into account pedagogical knowledge on the didactic domain as well as statistic information on both the student’s knowledge degree and learning preferences. Pedagogical information is described by means of an innovative methodology suitable for effective and efficient course generation and personalization. Moreover, statistic information can be collected and exploited by the system in order to better describe the student’s preferences and learning performances. Learning material is chosen by the system matching the student’s learning preferences with the learning material type, following a pedagogical approach suggested by Felder and Silverman. The paper discusses how automatic learning material personalization makes it possible to facilitate distance learning access to both able-bodied and disabled people. Results from the Diogene and Intraserv evaluation are reported and discussed.     

Simulating Emergent Properties of Coordination in Maude: the Collective Sort Case

Recent coordination languages and models are moving towards the application of techniques coming from the research context of complex systems: adaptivity and self-organization are exploited in order to tackle the openness, dynamism and unpredictability of today's distributed systems. In this area, systems are to be described using stochastic models, and simulation is a valuable tool both for analysis and design. Accordingly, in this work we focused on modelling and simulating emergent properties of coordination techniques.We first develop a framework acting as a general-purpose engine for simulating stochastic transition systems, built as a library for the Maude term rewriting system. We then evaluate this tool to a coordination problem called collective sort, where autonomous agents move tuples across different tuple spaces according to local criteria, and resulting in the emergence of the complete clustering property.     

Santolina pinnata Viv. Exerts Promising Antitumor Activity against Breast Cancer Cells and Anti-Inflammatory Effects in LPS-Stimulated RAW 264.7 Cells

Cancer is one of the largest causes of mortality in the world, and due to its incidence, the discovery of novel anticancer drugs is of great importance. Many successful anticancer drugs used in clinical practices are derived from natural products. The genus Santolina is a group of species distributed in the Mediterranean area and used in traditional medicine for their biological properties. The aim of this work was to investigate, for the first time, the multi-target biological potential of Italian Santolina pinnata in relation to their chemical profile, by which an interesting natural source of valuable phytochemicals endowed with anticancer and anti-inflammatory features could be assessed. n-Hexane (EHSP) and methanol (EMSP) extracts were investigated by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) and ultra-high-performance liquid chromatography (UHPLC), respectively. Anti-proliferative activity was analyzed on MCF-7 and MDA-MB-231 breast cancer cells, as well as on non-tumorigenic MCF-10A cells, by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Apoptotic death was assessed by comet assay. Cell motility and invasive features were examined in highly invasive MDA-MB-231 by wound-healing scratches, while, in both breast cancer cell lines, by gel-zymography experiments. The anti-inflammatory potential was analyzed by nitric oxide (NO) production and the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) staining experiments in bacterial lipopolysaccharides (LPS) which stimulated RAW 264.7 cells. EHSP and EMSP extracts exhibited anticancer activity against breast cancer cells, promoting apoptotic death, as well as decreasing cell migration and invasive behaviours. The highest activity (IC₅₀ of 15.91 μg/mL) was detected against MDA-MB-231 cells, a highly invasive breast cancer cell line. Both extracts were also able to promote anti-inflammatory effects (IC₅₀ values ranging from 27.5 to 61.14 μg/mL), as well as to reduce NO levels by inducing inhibitory effects on NF-κB nuclear translocation in LPS-stimulated RAW 264.7 cells. The different biological behaviours found between the extracts could be related to their different chemical compositions. Herein, the multi-target biological potential of S. pinnata in inducing antitumor and anti-inflammatory effects was comprehensively demonstrated. These findings will provide important stepping-stones for further investigations and may lead to the development of highly effective S. pinnata extract-based treatments for breast cancer and inflammatory processes.     

The PINK1 p.Asn521Thr Variant Is Associated with Earlier Disease Onset in GRN/C9orf72 Frontotemporal Lobar Degeneration

Genetic frontotemporal lobar degeneration (FTLD) is characterized by heterogeneous phenotypic expression, with a disease onset highly variable even in patients carrying the same mutation. Herein we investigated if variants in lysosomal genes modulate the age of onset both in FTLD due to GRN null mutations and C9orf72 expansion. In a total of 127 subjects (n = 74 GRN mutations and n = 53 C9orf72 expansion carriers), we performed targeted sequencing of the top 98 genes belonging to the lysosomal pathway, selected based on their high expression in multiple brain regions. We described an earlier disease onset in GRN/C9orf72 pedigrees in subjects carrying the p.Asn521Thr variant (rs1043424) in PTEN-induced kinase 1 (PINK1), a gene that is already known to be involved in neurodegenerative diseases. We found that: (i) the PINK1 rs1043424 C allele is significantly associated with the age of onset; (ii) every risk C allele increases hazard by 2.11%; (iii) the estimated median age of onset in homozygous risk allele carriers is 10–12 years earlier than heterozygous/wild type homozygous subjects. A replication study in GRN/C9orf72 negative FTLD patients confirmed that the rs1043424 C allele was associated with earlier disease onset (−5.5 years in CC versus A carriers). Understanding the potential mechanisms behind the observed modulating effect of the PINK1 gene in FTLD might prove critical for identifying biomarkers and/or designing drugs to modify the age of onset, especially in GRN/C9orf72-driven disease.     

Translational Implications for Radiosensitizing Strategies in Rhabdomyosarcoma

Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma of childhood and adolescence that includes FP-RMS, harboring the fusion oncoprotein PAX3/7-FOXO1 and FN-RMS, often mutant in the RAS pathway. Risk stratifications of RMS patients determine different prognostic groups and related therapeutic treatment. Current multimodal therapeutic strategies involve surgery, chemotherapy (CHT) and radiotherapy (RT), but despite the deeper knowledge of response mechanisms underpinning CHT treatment and the technological improvements that characterize RT, local failures and recurrence frequently occur. This review sums up the RMS classification and the management of RMS patients, with special attention to RT treatment and possible radiosensitizing strategies for RMS tumors. Indeed, RMS radioresistance is a clinical problem and further studies aimed at dissecting radioresistant molecular mechanisms are needed to identify specific targets to hit, thus improving RT-induced cytotoxicity.