Intranucleolar localization of DNA topoisomerase IIalpha is a distinctive feature of necrotic, but not of apoptotic, Jurkat T-cells

Two distinct types of cell death have been described: apoptosis and necrosis. However, it is becoming increasingly clear that the differences between these two types are far less numerous than initially thought. Morphological analyses might provide important information to distinguish apoptotic from necrotic samples. We recently reported that in necrotic, but not apoptotic, HL-60 human myeloid leukaemia cells, the nuclear protein topoisomerase IIalpha concentrated in nucleoli. In order to ascertain whether or not this phenomenon was restricted to a peculiar cell type or could be detected also in cells of lymphoid lineage, we performed an investigation aimed at defining the localization of topoisomerase IIalpha in apoptotic and necrotic Jurkat human T lymphoblastoid cells. Immunofluorescence staining demonstrated that topoisomerase IIalpha was excluded from the condensed chromatin of apoptotic cells, whereas in necrotic cells it was localized in discrete nuclear dots. Immuno-electron microscopy analysis showed that topoisomerase IIalpha was undetectable in nucleoli of normal and apoptotic cells, whereas it was present in the nucleolus of necrotic cells irrespectively of the type of inducer used (ethanol, H(2)O(2), HgCl(2)). Taken together, our findings identify topoisomerase IIalpha as a potential morphological marker useful to discriminate between apoptotic and necrotic cells.     

Virtual agent multimodal mimicry of humans

This work is about multimodal and expressive synthesis on virtual agents, based on the analysis of actions performed by human users. As input we consider the image sequence of the recorded human behavior. Computer vision and image processing techniques are incorporated in order to detect cues needed for expressivity features extraction. The multimodality of the approach lies in the fact that both facial and gestural aspects of the user’s behavior are analyzed and processed. The mimicry consists of perception, interpretation, planning and animation of the expressions shown by the human, resulting not in an exact duplicate rather than an expressive model of the user’s original behavior.     

Nanostructured TiO2-based mixed metal oxides prepared using microemulsions for carbon monoxide detection

Nanostructured powders of Nb-doped TiO₂ (TN) and SnO₂ mixed with Nb-doped TiO₂ in two different atomic ratios—10 to 1 (TSN 101) and 1 to 1 (TSN 11)—were synthesized using the reverse micelle microemulsion of a nonionic surfactant (brine solution/1-hexanol/Triton X-100/cyclohexane). The powders were characterized by transmission electron microscopy (TEM) and X-ray diffraction (XRD). Thick films were fabricated for gas sensors and characterized by XRD analysis and field emission scanning electron microscopy (FE-SEM). The effects of the film morphology and firing temperature in the range 650–850 °C on CO sensitivity were studied. The best gas response, expressed as the ratio between the resistance in air and the resistance under gas exposure (R _air/R _gas), was measured for TSN 11 at 11 for 1,000 ppm CO exposure. All types of sensors showed good thermal stability. The electrochemical impedance spectroscopy (EIS) measurements were performed in different gas atmospheres (air, O₂, CO and NO₂) to better understand the electrical properties of the nanostructured mixed metal oxides.     

Fabrication of photoactive heterostructures based on quantum dots decorated with Au nanoparticles

Silica based multifunctional heterostructures, exhibiting near infrared (NIR) absorption (650–1200 nm) and luminescence in the visible region, represent innovative nanosystems useful for diagnostic or theranostic applications. Herein, colloidal synthetic procedures are applied to design a photoactive multifunctional nanosystem. Luminescent silica (SiO₂) coated quantum dots (QDs) have been used as versatile nanoplatforms to assemble on their surface gold (Au) seeds, further grown into Au spackled structures. The synthesized nanostructures combine the QD emission in the visible region, and, concomitantly, the distinctive NIR absorption of Au nanodomains. The possibility of having multiple QDs in a single heterostructure, the SiO₂ shell thickness, and the extent of Au deposition onto SiO₂ surface have been carefully controlled. The work shows that a single QD entrapped in 16 nm thick SiO₂ shell, coated with Au speckles, represents the most suitable geometry to preserve the QD emission in the visible region and to generate NIR absorption from metal NPs. The resulting architectures present a biomedical potential as an effective optical multimodal probes and as promising therapeutic agents due to the Au NP mediated photothermal effect.     

Usage of SoS methodologies in production system design

Production systems design is a multifaceted task, due to a variety of aspects such as the mutual interdependency between the sub-systems, the variety of configurations and alternative system control strategies, the multiple managerial and “soft” aspects that cast an influence on the behavior of the system. A reasonable number of modeling tools can be applied to production system design, but they tend to divide the problem into unconnected sub-problems whose individual solutions may result in a poor global one. This is despite the fact that production design encompasses all aspects of manufacturing operations, and needs a systemic approach, as clearly shown in practitioner-oriented literature. This paper proposes to apply the “System of Systems” approach to production system design in order to represent their main aspects and support the rational definition of the path leading from corporate strategy to system (re)design.     

Monophasic Nanostructured Powders of Niobium, Tantalum, and Hafnium Carbonitrides Synthesized by a Mechanically Induced Self-Propagating Reaction

Niobium, tantalum, and hafnium carbonitride nanopowders were obtained by high-energy ball milling of elemental metal and graphite in a nitrogen atmosphere. By adjusting milling parameters, a mechanically induced self-propagating reaction was observed during the formation of carbonitride phases. The stoichiometry of transition metal carbonitrides was controlled by fixing the starting metal-to-carbon molar ratio. Ignition of the reactant mixture was determined by the combined effect of nitrogen and carbon, gas, and solid reagents, respectively. The composition and lattice parameters were determined by X-ray diffraction measurements. Microstructural characterization showed a nanophase structure with a homogeneous chemical composition.     

Apoptotic DNA fragmentation can be revealed in situ: An ultrastructural approach

A common pattern of apoptotic death is DNA cleavage, initially producing large fragments (50 kbp), followed by the production of nucleosomic/oligonucleosomic fragments. Nevertheless, apoptosis without DNA fragmentation, at least of the nucleosomic type, has been reported. To investigate the spatial relationship between DNA cleavage and chromatin condensation, we applied the TUNEL technique to the ultrastructural analysis of apoptotic cells. A modified method, utilizing a gold‐conjugated antidigoxigenin antibody, was carried out on U937 versus Molt‐4 cells, both exposed to UVB radiation or staurosporine treatment. Gold particle density in the different domains of apoptotic cells was evaluated by a four‐way ANOVA test. Gold labelling was more strongly localised in condensed chromatin than in the diffuse chromatin. U937 cells, which evidenced in vitro oligonucleosomic fragmentation after both UVB and staurosporine treatments, revealed a significantly higher gold particle density, when compared with Molt‐4, which did not show, on the other hand, oligonucleosomic cleavage even in the presence of ≤50 kbp cleavage. Thus, a correlation between DNA fragment sizes and gold particle density appears. TUNEL applied to electron microscopy is an effective approach to study the relationship between apoptotic chromatin condensation and DNA cleavage. Both these events indeed appear in the apoptotic nucleus, but their reciprocal correlation is still greatly unknown. Microsc. Res. Tech. 2009. © 2009 Wiley‐Liss, Inc.     

Critical Review of the Evolution of Extracellular Vesicles’ Knowledge: From 1946 to Today

Extracellular vesicles (EVs) are a family of particles/vesicles present in blood and body fluids, composed of phospholipid bilayers that carry a variety of molecules that can mediate cell communication, modulating crucial cell processes such as homeostasis, induction/dampening of inflammation, and promotion of repair. Their existence, initially suspected in 1946 and confirmed in 1967, spurred a sharp increase in the number of scientific publications. Paradoxically, the increasing interest for EV content and function progressively reduced the relevance for a precise nomenclature in classifying EVs, therefore leading to a confusing scientific production. The aim of this review was to analyze the evolution of the progress in the knowledge and definition of EVs over the years, with an overview of the methodologies used for the identification of the vesicles, their cell of origin, and the detection of their cargo. The MISEV 2018 guidelines for the proper recognition nomenclature and ways to study EVs are summarized. The review finishes with a “more questions than answers” chapter, in which some of the problems we still face to fully understand the EV function and potential as a diagnostic and therapeutic tool are analyzed.     

Ketoprofen Spray-dried Microspheres Based on Eudragit® RS and RL: Study of the Manufacturing Parameters

The preparation of ketoprofen spray-dried microspheres can be affected by the long drug recrystallization time. Polymer type and drug–polymer ratio as well as manufacturing parameters affect the preparation. The purpose of this work was to evaluate the possibility to obtain ketoprofen spray-dried microspheres using the Eudragit® RS and RL; the influence of the spray-drying parameters on morphology, dimension, and physical stability of microspheres was studied. Ketoprofen microspheres based on Eudragit® blend can be prepared by spray-drying and the nebulization parameters do not influence significantly particle properties; nevertheless, they can be affected by drying and storage methods. No effect of the container material is found.