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Scientific investigations of artworks are crucial in terms of preservation since they provide a measurable evaluation of the materials and the state of conservation. This is the case of Antonello da Messina's painting "Ecce Homo": its delicate state of conservation, with the need for constant monitoring, required a broad and in-depth diagnostic campaign to support the restorers. The project was carried out entirely in situ using non-invasive cutting-edge techniques and proposes a multimodal and data-centric approach, integrating 3D and 2D methodologies. The surface irregularities and the support were analysed with a structured-light 3D scanner and X-ray tomography. The painting materials were investigated with X-ray fluorescence scanning (MA-XRF) and reflectance hyperspectral imaging (HSI). Primarily, the data were jointly used for a scientific scope and provided new knowledge of the painting in terms of materials and painting techniques. In addition, two web-based interactive platforms were developed: one to provide restorers and experts with a new perspective of the hidden geometries of the painting, and the other targeted at the general public for dissemination purposes. The results of the Ecce Homo scientific analysis were exhibited, using a touch-screen interface, and developed for different user levels, from adults to kids. [ABSTRACT FROM AUTHOR]

Current approaches for the treatment of chronic lymphocytic leukemia (CLL) have greatly improved the prognosis for survival, but some patients remain refractive to these therapeutic regimens. Hence, in addition to reducing the long-term sideeffects of therapeutics for all leukemia patients, there is an urgent need for novel therapeutic strategies for difficult-to-treat leukemia cases. Due to the cytotoxicity of drugs, the major challenge currently is to deliver the therapeutic agents to neoplastic cells while preserving the viability of non-malignant cells. In this study, we propose a therapeutic approach in which high doses of hydroxychloroquine and chlorambucil were loaded into biodegradable polymeric nanoparticles coated with an anti-CD20 antibody.We first demonstrated the ability of the nanoparticles to target and internalize in tumor B-cells. Moreover, these nanoparticles could kill not only p53-mutated/deleted leukemia cells expressing a low amount of CD20, but also circulating primary cells isolated from chronic lymphocytic leukemia patients. The safety of these nanoparticles was also demonstrated in healthy mice, and their therapeutic effects were shown in a new model of aggressive leukemia. These results showed that anti-CD20 nanoparticles containing hydroxychloroquine and chlorambucil can be effective in controlling aggressive leukemia and provided a rationale for adopting this approach for the treatment of other B-cell disorders. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Mast cells (MC) play a key role in triggering the inflammatory process and share some functions with professional phagocytes. It is not clear whether or not the phagocytic process in MC follows the same route and has the same meaning of that of professional phagocytes. Herein we analyze in detail the structure of the phagosome in rat peritoneal mast cells (RPMC). The ultrastructural analysis of the phagosome, containing either model particles or bacteria, reveals that these vacuoles are very tight, and in several areas, their membrane seems to have dissolved. RPMC express NOD1 and NOD2 proteins whose role is to recognize intracellular foreign components and induce the production of pro-inflammatory mediators. Following Escherichia coli ingestion, both these molecules are found on the phagosome membrane and on ingested pathogens, together with phagosome maturation markers. These findings suggest that in RPMC the ingested cargo can, through interruptions of the phagosome membrane, interact directly with NODs, which act as switches in the process of cytokine production. [ABSTRACT FROM AUTHOR]

Mast cells (MCs) reside in tissues that are common targets of Candida spp. infections, and can exert bactericidal activity, but little is known about their fungicidal activity. MCs purified from rat peritoneum (RPMC) and a clinical isolate of C. albicans, were employed. Ingestion was evaluated by flow cytometry (FACS) and optical microscopy. The killing activity was assayed by FACS analysis and by colony forming unit method. RPMC degranulation was evaluated by β-hexosaminidase assay and phosphatidylserine externalization by FACS. Phagocytosing RPMC were also analyzed by transmission electron microscopy. Herein, we show that the killing of C. albicans by RPMC takes place in the extracellular environment, very likely through secreted granular components. Ultrastructural analysis of the ingestion process revealed an unusual RPMC- C. albicans interaction that could allow fungal survival. Our findings indicate that MCs have a positive role in the defense mechanism against Candida infections and should be included among the cell types involved in host-defense against this pathogen. [ABSTRACT FROM AUTHOR]

In older persons, vitamin D insufficiency and a subclinical chronic inflammatory status frequently coexist. Vitamin D has immune-modulatory and in vitro anti-inflammatory properties. However, there is inconclusive evidence about the anti-inflammatory role of vitamin D in older subjects. Thus, we investigated the hypothesis of an inverse relationship between 25-hydroxyvitamin D (25(OH)D) and inflammatory markers in a population-based study of older individuals. After excluding participants with high-sensitivity C-reactive protein (hsCRP) ≥ 10 mg/dl and those who were on chronic anti-inflammatory treatment, we evaluated 867 older adults ≥65 years from the InCHIANTI Study. Participants had complete data on serum concentrations of 25(OH)D, hsCRP, tumor necrosis factor (TNF)-α, soluble TNF-α receptors 1 and 2, interleukin (IL)-1β, IL-1 receptor antagonist, IL-10, IL-18, IL-6, and soluble IL-6 receptors (sIL6r and sgp130). Two general linear models were fit (model 1-adjusted for age, sex, and parathyroid hormone (PTH); model 2-including covariates of model 1 plus dietary and smoking habits, physical activity, ADL disability, season, osteoporosis, depressive status, and comorbidities). The mean age was 75.1 ± 17.1 years ± SD. In model 1, log(25OH-D) was significantly and inversely associated with log(IL-6) ( β ± SE = −0.11 ± 0.03, p = <0.0001) and log (hsCRP) ( β ± SE = −0.04 ± 0.02, p = 0.04) and positively associated with log(sIL6r) ( β ± SE = 0.11 ± 0.04, p = 0.003) but not with other inflammatory markers. In model 2, log (25OH-D) remained negatively associated with log (IL-6) ( β ± SE = −0.10 ± 0.03, p = 0.0001) and positively associated with log(sIL6r) ( β ± SE = 0.11 ± 0.03, p = 0.004) but not with log(hsCRP) ( β ± SE = −0.01 ± 0.03, p = 0.07). 25(OH)D is independently and inversely associated with IL-6 and positively with sIL6r, suggesting a potential anti-inflammatory role for vitamin D in older individuals. [ABSTRACT FROM AUTHOR]

We have hypothesized that mast cells (MC) and eosinophils (Eos), the main effectors of allergy, can form an effector unit. These cells co-exist in the inflamed tissues during the late and chronic stages of allergy and have been shown to be capable of influencing each other's survival and activity via soluble mediators. We have recently described couples of receptor-ligands that are expressed on either/both of these cells and that imply a physical interaction. In this study, we have investigated the existence of short-term (60 min) in vitro interactions between human peripheral blood Eos and cord-blood-derived MC by transmission electron microscopy. We have found that MC and Eos adhere to each other; the lipid body content and the granule morphology of co-cultured MC and Eos, respectively, are altered, and the level of Eos peroxidase (EPO) released by co-cultured Eos is elevated. Moreover, the transfer of EPO from Eos to MC and tryptase from MC to Eos has been observed. Our results thus indicate that, when co-cultured, MC and Eos show signs of physical contact and of reciprocal activation. This is the first in vitro evidence of functional physical interactions between human MC and Eos, interactions that might also occur in vivo during allergic diseases. [ABSTRACT FROM AUTHOR]

Reports on a study conducted by researchers in Italy, which shows that human neutrophils and promyelocitic cells express neuronal calcium sensor-1, a calcium binding protein involved in exocytosis in various cell types. Association of neuronal calcium sensor-1 with azurophilic granules; Role of neuronal calcium sensor-1 in the calcium-dependent secretion of azurophilic granules.

The microbicidal activity of phagocytes depends on intraphagosomal secretion (i.e., the degranulation process), during which the content of the secretory granules is discharged into the phagosome to kill ingested microorganisms. The availability of a reliable assay to quantify the extent of degranulation can be an important tool to gain a deeper insight into the mechanism of bacterial processing in phagocytes and into the transmembrane signaling that leads to granule–phagosome fusion. In an early study, Baehner et al. (25) showed by using the peroxidase chromogenic substrate DAB that the occurence of intraphagosomal MPO release in human neutrophils. Starting from that finding and from that of Herzog and Fahimi (26), who reported the quantitative evaluation of peroxidase-mediated DAB oxidation, we set up a method for measuring MPO intraphagosomal release in human neutrophils. The method is based on the passive engulfment of DAB together with the phagocytosable particle. Inside the vacuole, this substrate is oxidized by MPO released from the azurophilic granules. The colorimetrical evaluation of the amount of DAB oxidized allows for cheap, rapid quantification of MPO intraphagosomal secretion in whole cells. Using this method, we show that the degranulation process, involving azurophilic granules, can be monitored carefully during phagocytosis. It takes place after the ingestion of zymosan particles opsonized with normal human serum, as well as during IgG-mediated phagocytosis and under conditions where β2 integrins are blocked. However our findings also show that the extent of intraphagosomal secretion depends on either the extent of opsonization or the type of receptor engaged during the phagocytic event. [ABSTRACT FROM AUTHOR]

The surface composition of the azurophilic granule of human neutrophils is largely unknown. To understand the role played by the membrane components of these organelles a simple methodology for their disruption and the isolation of suitable amounts of granule membranes is required. The method of disruption of azurophilic granules available up to now consists of seven steps of freezing-thawing and sonication, a method time-consuming when large amount of granule needs to be processed. In this paper we describe a simple, one-step method based on the granule damaging capacity of Percoll which is suitable for the disruption of large amounts of azurophilic granules. The components of granule lysates obtained with this method were resolved on sucrose gradients. Two major bands were obtained, which are shown to be different from each other judging from their content of selective markers. [ABSTRACT FROM AUTHOR]

The so-called amphibole asbestos fibers are enriched with mineral iron ions, able to stimulate ROS production. We recently reported that crocidolite asbestos was able to interact with the cell membranes of Xenopus laevis oocytes, to alter their electrical membrane properties. Here, we found that applied iron ions (Fe3+) or H2O2 (for ROS generation) mimicked these effects, suggesting that at least one effect of iron-containing asbestos fiber exposure was mediated by ROS production. Furthermore, combined Fe3+ and H2O2 acted synergistically, producing a membrane effect stronger than that induced by these factors alone. Similar to crocidolite, these changes peaked within 30 minutes of incubation and vanished almost completely after 120 min. However, in the presence of cytochalasin D, which inhibits membrane actin repair mechanisms, crocidolite or applied Fe3+/H2O2 invariably produced oocyte cell death. While the electrophysiological modifications induced by crocidolite suggested a modification of an intrinsic chloride ion channel, the morphological appearance of the treated oocytes also indicated the formation of membrane "pores"; the effects of asbestos exposure may therefore consist of multiple (not necessarily exclusive) underlying mechanisms. In conclusion, using Xenopus oocytes allowed us for the first time, to focus on a specific membrane effect of crocidolite asbestos exposure, which deserves to be tested also on human lung cell lines. Much available evidence suggests that asbestos fibers damage cells through the production of ROS. Our present data confirm that crocidolite fibers can indeed trigger ROS-mediated damaging effects in the oocyte cell membrane, provided iron ions and H2O2 are available for ROS production. [ABSTRACT FROM AUTHOR]