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Type 1 diabetes mellitus (T1D) is a chronic disease characterized by an autoimmune destruction of insulin-producing β-pancreatic cells. Although many advances have been achieved in T1D treatment, current therapy strategies are often unable to maintain perfect control of glycemic levels. Several studies are searching for new and improved methodologies for expansion of β-cell cultures in vitro to increase the supply of these cells for pancreatic islets replacement therapy. A promising approach consists of differentiation of stem cells into insulin-producing cells (IPCs) in sufficient number and functional status to be transplanted. Differentiation protocols have been designed using consecutive cytokines or signaling modulator treatments, at specific dosages, to activate or inhibit the main signaling pathways that control the differentiation of induced pluripotent stem cells (iPSCs) into pancreatic β-cells. Here, we provide an overview of the current approaches and achievements in obtaining stem cell-derived β-cells and the numerous challenges, which still need to be overcome to achieve this goal. Clinical translation of stem cells-derived β-cells for efficient maintenance of long-term euglycemia remains a major issue. Therefore, research efforts have been directed to the final steps of in vitro differentiation, aiming at production of functional and mature β-cells and integration of interdisciplinary fields to generate efficient cell therapy strategies capable of reversing the clinical outcome of T1D.

Mosquito saliva facilitates blood feeding through the anti-haemostatic, anti-inflammatory and immunomodulatory properties of its proteins. However, the potential contribution of non-coding RNAs to host manipulation is still poorly understood. We analysed small RNAs from Aedes aegypti saliva and salivary glands and show here that chikungunya virus-infection triggers both the siRNA and piRNA antiviral pathways with limited effects on miRNA expression profiles. Saliva appears enriched in specific miRNA subsets and its miRNA content is well conserved among mosquitoes and ticks, clearly pointing to a non-random sorting and occurrence. Finally, we provide evidence that miRNAs from Ae. aegypti saliva may target human immune and inflammatory pathways, as indicated by prediction analysis and searching for experimentally validated targets of identical human miRNAs. Overall, we believe these observations convincingly support a scenario where both proteins and miRNAs from mosquito saliva are injected into vertebrates during blood feeding and contribute to the complex vector–host–pathogen interactions.

During blood feeding haematophagous arthropods inject into their hosts a cocktail of salivary proteins whose main role is to counteract host haemostasis, inflammation and immunity. However, animal body fluids are known to also carry miRNAs. To get insights into saliva and salivary gland miRNA repertoires of the African malaria vector Anopheles coluzzii we used small RNA-Seq and identified 214 miRNAs, including tissue-enriched, sex-biased and putative novel anopheline miRNAs. Noteworthy, miRNAs were asymmetrically distributed between saliva and salivary glands, suggesting that selected miRNAs may be preferentially directed toward mosquito saliva. The evolutionary conservation of a subset of saliva miRNAs in Anopheles and Aedes mosquitoes, and in the tick Ixodes ricinus, supports the idea of a non-random occurrence pointing to their possible physiological role in blood feeding by arthropods. Strikingly, eleven of the most abundant An. coluzzi saliva miRNAs mimicked human miRNAs. Prediction analysis and search for experimentally validated targets indicated that miRNAs from An. coluzzii saliva may act on host mRNAs involved in immune and inflammatory responses. Overall, this study raises the intriguing hypothesis that miRNAs injected into vertebrates with vector saliva may contribute to host manipulation with possible implication for vector-host interaction and pathogen transmission.

Extraction of the cardiac surfaces of interest from multi-detector computed tomographic (MDCT) data is a pre-requisite step for cardiac analysis, as well as for image guidance procedures. Most of the existing methods need manual corrections, which is time-consuming. We present a fully automatic segmentation technique for the extraction of the right ventricle, left ventricular endocardium and epicardium from MDCT images. The method consists in a 3D level set surface evolution approach coupled to a new stopping function based on a multiscale directional second derivative Gaussian filter, which is able to stop propagation precisely on the real boundary of the structures of interest. We validated the segmentation method on 18 MDCT volumes from healthy and pathologic subjects using manual segmentation performed by a team of expert radiologists as gold standard. Segmentation errors were assessed for each structure resulting in a surface-to-surface mean error below 0.5 mm and a percentage of surface distance with errors less than 1 mm above 80%. Moreover, in comparison to other segmentation approaches, already proposed in previous work, our method presented an improved accuracy (with surface distance errors less than 1 mm increased of 8-20% for all structures). The obtained results suggest that our approach is accurate and effective for the segmentation of ventricular cavities and myocardium from MDCT images.

Colorectal cancers (CRCs) often show a dense infiltrate of cytokine-producing immune/inflammatory cells. The exact contribution of each immune cell subset and cytokine in the activation of the intracellular pathways sustaining CRC cell growth is not understood. Herein, we isolate tumor-infiltrating leukocytes (TILs) and lamina propria mononuclear cells (LPMCs) from the tumor area and the macroscopically unaffected, adjacent, colonic mucosa of patients who underwent resection for sporadic CRC and show that the culture supernatants of TILs, but not of LPMCs, potently enhance the growth of human CRC cell lines through the activation of the oncogenic transcription factors signal transducer and activator of transcription 3 (STAT3) and nuclear factor-kappa B (NF-kB). Characterization of immune cell complexity of TILs and LPMCs reveals no differences in the percentages of T cells, natural killer T cells, natural killer (NK) cells, macrophages and B cells. However, T cells from TILs show a functional switch compared with those from LPMCs to produce large amounts of T helper type 17 (Th17)-related cytokines (that is, interleukin-17A (IL-17A), IL-17F, IL-21 and IL-22), tumor necrosis factor-alpha (TNF-alpha) and IL-6. Individual neutralization of IL-17A, IL-17F, IL-21, IL-22, TNF-alpha or IL-6 does not change TIL-derived supernatant-driven STAT3 and NF-kB activation, as well as their proproliferative effect in CRC cells. In contrast, simultaneous neutralization of both IL-17A and TNF-alpha, which abrogates NF-kB signaling, and IL-22 and IL-6, which abrogates STAT3 signaling, reduces the mitogenic effect of supernatants in CRC cells. IL-17A, IL-21, IL-22, TNF-alpha and IL-6 are also produced in excess in the early colonic lesions in a mouse model of sporadic CRC, associated with enhanced STAT3/NF-kB activation. Mice therapeutically given BP-1-102, an orally bioavailable compound targeting STAT3/NF-kB activation and cross-talk, exhibit reduced colon tumorigenesis and diminished expression of STAT3/NF-kB-activating cytokines in the neoplastic areas. These data suggest that strategies aimed at the cotargeting of STAT3/NF-kB activation and interaction between them might represent an attractive and novel approach to combat CRC.

The importance of autophagy in mechanisms underlying inflammation has been highlighted. Downstream effects of the bacterial sensor NOD2 include autophagy induction. Recently, a relationship between defects in autophagy and adherent/invasive Escherichia coli (AIEC) persistence has emerged. The present study aims at investigating the interplay between autophagy, NOD2 and AIEC bacteria and assessing the expression level of autophagic proteins in intestinal biopsies of pediatric patients with inflammatory bowel disease (IBD). A human epithelial colorectal adenocarcinoma (Caco2) cell line stably over-expressing NOD2 was produced (Caco2NOD2). ATG16L1, LC3 and NOD2 levels were analysed in the Caco2 cell line and Caco2NOD2 after exposure to AIEC strains, by western blot and immunofluorescence. AIEC survival inside cells and TNFα, IL-8 and IL-1βmRNA expression were analysed by gentamicin protection assay and real time PCR. ATG16L1 and LC3 expression was analyzed in the inflamed ileum and colon of 28 patients with Crohn’s disease (CD), 14 with ulcerative colitis (UC) and 23 controls by western blot. AIEC infection increased ATG16L1 and LC3 in Caco2 cells. Exposure to AIEC strains increased LC3 and ATG16L1 in Caco2 overexpressing NOD2, more than in Caco2 wild type, while a decrease of AIEC survival rate and cytokine expression was observed in the same cell line. LC3 expression was increased in the inflamed colon of CD and UC children. The NOD2-mediated autophagy induction is crucial to hold the intramucosal bacterial burden, especially towards AIEC, and to limit the resulting inflammatory response. Autophagy is active in inflamed colonic tissues of IBD pediatric patients.

Maturity index (MI), based on nematode life strategies, has been proposed in 1990 to assess the possible variations of the terrestrial and freshwater nematode assemblages induced by anthropogenic activities. It was subsequently applied also to marine ecosystems and, even if comparatively not yet very popular, it offers a good method to assess the ecological quality in relation to a wide range of anthropogenic drivers. However, few data are available on its response to physical stress, a key factor especially in the coastal areas. In this study, marine nematode genera from two study cases carried out in Maldives are used to test both MI and life strategy traits (i.e., c-p classes) for detecting the effects of physical disturbance. The results confirm that nematodes are well adapted to physical stress showing a general high rate of recovery. C-p scaling and MI did not seem to be appropriate for revealing this disturbance type probably because there are no empirical evidences on the life strategy of several genera, and a possible differential response to various disturbance types may be hypothesized.

We are developing a Lumped Element Kinetic Inductance Detector (LEKID) array able to operate in the W-band (75-110 GHz) in order to perform ground-based Cosmic Microwave Background (CMB) and mm-wave astronomical observations. The W-band is close to optimal in terms of contamination of the CMB from Galactic synchrotron, free-free, and thermal interstellar dust. In this band, the atmosphere has very good transparency, allowing interesting ground-based observations with large (>30 m) telescopes, achieving high angular resolution (, 16th International Workshop on Low Temperature Detectors, Grenoble 20-24 July 2015, Journal of Low Temperature Physics, Accepted

The goal of the Cryogenic wide-Area Light Detectors with Excellent Resolution project is the development of light detectors with large active area and noise energy resolution smaller than 20 eV RMS using phonon-mediated kinetic inductance detectors. The detectors are developed to improve the background suppression in large-mass bolometric experiments such as CUORE, via the double read-out of the light and the heat released by particles interacting in the bolometers. In this work, we present the design and the fabrication process, starting from the silicon wafer arriving to the single chip. The Al thin films (40 nm) are evaporated on high-quality, high-resistivity (> $$10\,\mathrm{k \Omega }\,$$ cm) Si(100) substrates using an electron beam evaporator in a high-vacuum chamber. Detectors are patterned in direct-write mode, using electron beam lithography , positive tone resist poly-methyl methacrylate and lift-off process. Finally the wafer is diced into 20 $$\times $$ 20 mm $$^2$$ chips and assembled in a holder OFHC copper (oxygen-free high conductivity) using PTFE supports.

The present study analyzes the structure and dynamics of the forest landscape in a peri-urban area (Rome, Italy) during the city’s expansion from 1949 to 2008 using landscape metrics and change detection analysis of digital maps of the area (1500 km2). While urban settlements increased continuously from 6.5 to 27.5 % of the study area, woodlands changed less clearly, with a moderate increase (from 11.2 to 11.9 % of the total area) and a higher fragmentation as a consequence of Rome’s expansion. The structure of forest landscape changed along the urban-to-rural gradient with patch size increasing with the distance to the inner city in 1949 and substantial landscape homogeneity in 2008. The indicators proposed in this study inform dedicated measures for conserving forest and maintaining landscape diversity. Measures adopted in Rome’s forestation plan to counteract woodland fragmentation were analyzed and discussed. Based on the complex landscape dynamics found for Rome, an integrated multiscale planning approach targeting forest conservation is considered a key contribution to urban sustainability.

Anisakiasis is a fish-borne zoonosis caused by Anisakis spp. larvae. One challenging issue in the diagnosis of anisakiasis is the molecular detection of the etiological agent even at very low quantity, such as in gastric or intestinal biopsy and granulomas. Aims of this study were: 1) to identify three new cases of invasive anisakiasis, by a species-specific Real-time PCR probe assay; 2) to detect immune response of the patients against the pathogen. Parasite DNA was extracted from parasites removed in the three patients. The identification of larvae removed at gastric and intestinal level from two patients was first obtained by sequence analysis of mtDNA cox2 and EF1 α-1 of nDNA genes. This was not possible in the third patient, because of the very low DNA quantity obtained from a single one histological section of a surgically removed granuloma. Real-time PCR species-specific hydrolysis probe system, based on mtDNA cox2 gene, was performed on parasites tissue of the three cases. IgE, IgG4 and IgG immune response against antigens A. pegreffii by Immunoblotting assay was also studied. According to the mtDNA cox2 and the EF1 α − 1 nDNA sequence analysis, the larvae from stomach and intestine of two patients were assigned to A. pegreffii. The Real-time PCR primers/probe system, showed a fluorescent signal at 510 nm for A. pegreffii, in all the three cases. In Immunoblotting assay, patient CC1 showed IgE, IgG4 reactivity against Ani s 13-like and Ani s 7-like; patient CC2 revealed only IgG reactivity against Ani s 13-like and Ani s 7-like; while, the third patient showed IgE and IgG reactivity against Ani s 13-like, Ani s 7-like and Ani s 1-like. The Real-time PCR assay, a more sensitive method than direct DNA sequencing for the accurate and rapid identification of etiological agent of human anisakiasis, was successfully assessed for the first time. The study also highlights the importance to use both molecular and immunological tools in the diagnosis of human anisakiasis, in order to increase our knowledge about the pathological findings and immune response related to the infection by zoonotic species of the genus Anisakis.

Narrow Xi-pi+- and Xi-bar+pi+- resonances produced by quasi-real photons have been searched for by the COMPASS experiment at CERN. The study was stimulated by the recent observation of an exotic baryonic state decaying into Xi-pi-, at a mass of 1862 MeV, interpreted as a pentaquark. While the ordinary hyperon states Xi(1530)^0 and Xi-bar(1530)^0 are clearly seen, no exotic baryon is observed in the data taken in 2002 and 2003., Comment: 10 pages, 5 figures

Studies conducted in the prepubertal period showed that biomarkers of oxidative stress decreased with increasing age in normocholesterolemic children (NC), and, conversely, they are persistently high in hypercholesterolemic children (HC). Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is the most important cellular source of reactive oxygen species. No data have been reported concerning the behavior of age-related oxidative stress generated by NOX2, the catalytic subunit of NADPH oxidase, in children. We performed a cross-sectional study comparing oxidized low-density lipoprotein (ox-LDL), as a marker of oxidative stress, and NOX2 activity, as assessed by blood levels of soluble NOX2–derived peptide (sNOX2-dp), in a population of 250 children, including 125 NC and 125 HC. HC had higher sNOX2-dp (25.8 ± 16.1 pg/ml) and ox-LDL (23.6 ± 14.4 U/l) levels as compared with NC (15.4 ± 10.1 and 11.6 ± 6.4 pg/ml, respectively; P < 0.001). Multiple linear regression analysis showed that ox-LDL and total cholesterol were the only independent predictive variables associated with sNOX2-dp. In NC, ox-LDL and sNOX2-dp significantly decreased from the first to the second quintile of age. In HC, ox-LDL and sNOX2-dp levels did not show significant differences among quintiles of age. This study suggests that NOX2 contributes to generating ox-LDL during the early phase of life in NC and to the persistent high oxidative stress in the prepubertal period of HC.

Coral reefs are important hot spots of biodiversity, but despite the problems related to their conservation, studies on meiobenthic biodiversity are scarce. This paper provides a preliminary overview of the meiofauna and nematodes inhabiting coral sediments in the outer reef and lagoon habitats of the Maldives, and investigates their relationships with micro-habitat type. The abundance and community structure of the meiofauna and nematodes reveal significant differences between stations. Coral fragments and rubble from the outer reefs seem to act as a trap for the finest sediment, which might contribute to creating a heterogeneous micro-habitat suitable for both epifaunal (Epsilonematidae and Draconematidae genera) and sediment-dwelling (Tricoma, Richtersia, Ptycholaimellus and Molgolaimus) taxa. On the other side, the lagoon’s sediments are mainly colonized by dwelling taxa, probably due to the low-flow regime that enhances the deposition of organic matter and pelite fractions. A high level of diversity was recorded in both the habitats studied. Among the factors that probably determine the high diversity in the lagoon is the presence of small biogenic structures. These are characterized by a high degree of angularity, which might add more complexity to the habitat. The nematodes reveal an overlap in the taxonomic composition between the Maldives and other geographically distant areas, possibly supporting the existence of iso-communities that are typical of the coral degradation zone. Nematode trophic composition reflects differences in availability of food resources; epigrowth and non-selective deposit feeders are dominant in the lagoon, while selective deposit and epistrate feeders are dominant in the outer reefs.

Bacteria, meio- and macrofauna were investigated at different depths in a coastal area of the Central Adriatic Sea, yielding information about the composition and abundance of the benthic community. In particular, 14 nematode genera were recorded for the first time in the Upper Adriatic Sea. All communities resulted as being significantly different between inshore and offshore stations, especially when the season interaction was considered. Sediment grain size seemed to be the main natural variable, along with trophic resources, affecting the distribution and composition of these benthic components, whilst there was no clear evidence of competition for food sources and predatory pressure between the communities. Meiofauna appeared the most useful community for detecting disturbances and river influences. In particular, the lowest copepod abundance in the shallow waters seemed to be related to a greater anthropogenic disturbance inshore, whilst meiofaunal abundance and diversity together with the nematode maturity index suggest the influence of the Foglia and Metauro rivers and the small stream Arzilla.

We have analyzed data of the DISTO experiment on the exclusive pp to K^+ Lambda p process at T_p = 2.85$ GeV to search for a K^-pp (= X) nuclear bound state to be formed in the pp to K^+ + X reaction. The deviation spectra of the K^+ missing-mass MM (K^+) and Lambda p invariant-mass M(Lambda-p) with selection of large-angle proton emission revealed a structure with M_X = 2265 +- 2 MeV/c2 and Gamma_X = 118 +- 8$ MeV., Comment: 6 pages, 4 figues, Proceedings of EXA08

The bleaching event of 1998 caused widespread mortality on coral reefs in the Maldives. Nearly 10 years after the coral mass mortality, the state of Maldivian reefs was evaluated paying specific attention to three ecological stages, linked to the 3D structure of the coral community: young (high living hard coral cover), mature (a balance between living coral cover and loose sediment), and regressive (high amount of rubble and sand). The relative importance of three biogeomorphological descriptors (living hard corals; rubble and sand; coralline algae on rock) in the reef flat (2–7 m depth) and in the slope (7–18 m) of three reefscapes related to wave exposure was assessed. The role of wave energy in shaping ecological stages is different in the reef flat (early stages are found in low energy conditions) and in the slope (early stages are associated with high energy sites).