Three-dimensional apoptotic nuclear behavior analyzed by means of field emission in lens scanning electron microscope

Apoptosis is an essential biological function required during embryogenesis, tissue homeostasis, organ development and immune system regulation. It is an active cell death pathway involved in a variety of pathological conditions. During this process cytoskeletal proteins appear damaged and undergo an enzymatic disassembling, leading to formation of apoptotic features. This study was designed to examine the threedimensional chromatin behavior and cytoskeleton involvement, in particular actin re-modeling. HL-60 cells, exposed to hyperthermia, a known apoptotic trigger, were examined by means of a Field Emission in Lens Scanning Electron Microscope (FEISEM). Ultrastructural observations revealed in treated cells the presence of apoptotic patterns after hyperthermia trigger. In particular, three-dimensional apoptotic chromatin rearrangements appeared involving the translocation of filamentous actin from cytoplasm to the nucleus. FEISEM immunogold techniques showed actin labeling and its precise three-dimensional localization in the diffuse chromatin, well separated from the condensed one. The actin presence in dispersed chromatin inside the apoptotic nucleus can be considered an important feature, indispensable to permit the apoptotic machinery evolution

Flavonoid-inspired vascular disrupting agents: Exploring flavone-8-acetic acid and derivatives in the new century

Naturally occurring flavonoids are found as secondary metabolites in a wide number of plants exploited for both medicine and food and have long been known to be endowed with multiple biological activities, making them useful tools for the treatment of different pathologies. Due to the versatility of the scaffolds and the vast possibilities of appropriate decoration, they have also been regarded as fruitful sources of lead compounds and excellent chemical platforms for the development of bioactive synthetic compounds. Flavone-8-acetic acid (FAA) and 5,6-dimethylxanthone acetic acid (DMXAA) emerged for their antitumour potential due to the induction of cytokines and consequent rapid haemorrhagic necrosis of murine tumour vasculature, and different series of derivatives have been designed thereafter. Although the promising DMXAA failed in phase III clinical trials because of strict species-specificity, a boost in research came from the recent identification of the stimulator of interferon genes (STING), responsible for supporting tumoural innate immune responses, as a possible biological target. Consequently, in the last decade a renewal of interest for these flavonoid-based structures was noticed, and novel derivatives have been synthesised and evaluated for a deeper understanding of the molecular features needed for affecting human cells. Un-doubtedly, these natural-derived molecules deserve further investigation and still appear attractive in an anticancer perspective

Optimization of Esterase Production in Solid-State Fermentation of Agricultural Digestate

The continuous increase in biogas production poses the need for innovative applications for its by-products. Solid-state fermentation (SSF) has regained attention in the development of several products because of the possibility to use low-cost and easily available substrates, such as organic wastes. SSF represents a valuable process for agricultural digestate valorization in terms of enzyme production. In the present study, cellulase and esterase were produced by Trichoderma asperellum R on a digestate-based substrate in SSF, with esterase as the highest obtained activity. After assessing the effect of light on it, the esterase production in SSF was optimized using response surface methodology. The influence of substrate composition, temperature and humidity on the enzyme production was evaluated on two sets of data generated based on digestate concentration (50% and 70% w/w). The statistical analyses revealed that these parameters affected esterase production only when Trichoderma asperellum grew on substrate containing 50% w/w of digestate. The best esterase activity (264.6 mU/mg total protein) was achieved with the following optimized SSF parameters: 50% digestate, 50% fruits, 10% sawdust, 30 & DEG;C. The current finding of esterase production on digestate-based substrates makes the SSF method presented here a sustainable and completely circular technology

Impact of Extending Hard-Cheese Ripening : A Multiparameter Characterization of Parmigiano Reggiano Cheese Ripened up to 50 Months

Extending ripening of hard cheeses well beyond the traditional ripening period is becoming increasingly popular, although little is known about the actual evolution of their characteristics. The present work aimed at investigating selected traits of Parmigiano Reggiano cheese ripened for 12, 18, 24, 30, 40 and 50 months. Two cheeses per each ripening period were sampled. Although moisture constantly decreased and was close to 25% in 50-month cheeses, with a parallel increase in cheese hardness, several biochemical changes occurred involving the activity of both native and microbial enzymes. Capillary electrophoresis demonstrated degradation of \u3b1s1- and \u3b2-casein, indicating residual activity of both chymosin and plasmin. Similarly, continuous release of free amino acids supported the activity of peptidases deriving from lysed bacterial cells. Volatile flavor compounds, such as short-chain fatty acids and some derived ketones, alcohols and esters, evaluated by gas chromatography with solid-phase micro-extraction, accumulated as well. Cheese microstructure was characterized by free fat trapped in irregularly shaped areas within a protein network, with native fat globules being no longer visible. This study showed for the first time that numerous biochemical and structural variations still occur in a hard cheese at up to 50 months of aging, proving that the ripening extension deserves to be highlighted to the consumer and may justify a premium price

Object-based image analysis for historic maps classification

Heritage maps represent fundamental information for the study of the evolution of a region, especially in terms of landscape and ecologic features. Historical maps present two kinds of hurdle before they can be used in a modern GIS: they must be geometrically corrected to correspond to the datum in use and they must be classified to exploit the information they contain. This study deals the latter problem: the Historical Cadaster Map, created between 1851 and 1861, for the Trentino region in the North of Italy is available as a collection of maps in the ETRS89/UTM 32N datum. The map is a high resolution scan (230 DPI, 24 bit) of the original map and has been used in several ecological studies, since it provides detailed information not only about land property but also about land use. In the past the cadaster map has been manually digitized and for each area a set of attributes has been recorded. Since this approach is time consuming and prone to errors, automatic and semi-automatic procedures have been tested. Traditional image classification techniques, such as maximum likelihood classification, supervised or un-supervised, pixelwise and contextual, do not provide satisfactory results for many reasons: map colors are very variable within the same area, symbols and characters are used to identify cadaster parcels and locations, lines, drawn by hand on the original map, have variable thickness and colors. The availability of FOSS tools for the Object-based Image Analysis (OBIA) has made possible the application of this technique to the cadaster map. This paper describes the use of GRASS GIS and R for the implementation of the OBIA approach for the supervised classification of the historic cadaster map. It describes the determination of the optimal segments, the choice of their attributes and relevant statistics, and their classification. The result has been evaluated with respect to a manually digitized map using Cohens Kappa and the analysis of the confusion matrix. The result of the OBIA classification has also been compared to the classification of the same map using maximum likelihood classification, un-supervised and supervised, both pixelwise and contextual. The OBIA approach has provided very satisfactory results with the ability to automatically remove the background and symbols and characters, creating a ready to be used classified map. This study highlights the effectiveness of the OBIA processing chain available in the FOSS4G ecosystem, and in particular the added value of the interoperability between GRASS GIS and R

Cytokine Profiling in Myeloproliferative Neoplasms: Overview on Phenotype Correlation, Outcome Prediction, and Role of Genetic Variants

Among hematologic malignancies, the classic Philadelphia-negative chronic myeloproliferative neoplasms (MPNs) are considered a model of inflammation-related cancer development. In this context, the use of immune-modulating agents has recently expanded the MPN therapeutic scenario. Cytokines are key mediators of an auto-amplifying, detrimental cross-talk between the MPN clone and the tumor microenvironment represented by immune, stromal, and endothelial cells. This review focuses on recent advances in cytokine-profiling of MPN patients, analyzing different expression patterns among the three main Philadelphia-negative (Ph-negative) MPNs, as well as correlations with disease molecular profile, phenotype, progression, and outcome. The role of the megakaryocytic clone as the main source of cytokines, particularly in myelofibrosis, is also reviewed. Finally, we report emerging intriguing evidence on the contribution of host genetic variants to the chronic pro-inflammatory state that typifies MPNs