Our resistance to infection is critically dependent upon the ability of pattern recognition receptors to recognise microbial invasion and induce protective immune responses. One such family of receptors are the C-type lectins, which play central roles in antifungal immunity1. These receptors activate key effector mechanisms upon recognition of conserved fungal cell wall carbohydrates. However, several other immunologically active fungal ligands have been described, including melanin2,3, whose mechanisms of recognition remain largely undefined. Here we identify a C-type lectin receptor, Melanin sensing C-type Lectin receptor (MelLec), that plays an essential role in antifungal immunity through recognition of the naphthalene-diol unit of 1,8-dihydroxynaphthalene (DHN)-melanin. MelLec recognises melanin in conidial spores of Aspergillus fumigatus, as well as other DHN-melanised fungi and is ubiquitously expressed by CD31+ endothelial cells in mice. MelLec is also expressed by a sub-population of these cells in mice that co-express EpCAM and which were detected only in the lung and liver. In mouse models, MelLec was required for protection against disseminated infection with A. fumigatus. In humans, MelLec is also expressed by myeloid cells, and we identified a single nucleotide polymorphism of this receptor that negatively affected myeloid inflammatory responses and significantly increased susceptibility of stem-cell transplant recipients to disseminated Aspergillus infections. Thus MelLec is a receptor recognising an immunologically active component commonly found on fungi and plays an essential role in protective antifungal immunity in both mice and humans.

Various cells types, including stem and progenitor cells, can exchange complex information via plasma membrane-derived vesicles, which can carry signals both in their limiting membrane and lumen. Astrocytes, traditionally regarded as mere supportive cells, play previously unrecognized functions in neuronal modulation and are capable of releasing signalling molecules of different functional significance. In the present study, we provide direct evidence that human fetal astrocytes in culture, expressing the same feature as immature and reactive astrocytes, release membrane vesicles larger than the microvesicles described up to now. We found that these large vesicles, ranging from 1-5 to 8 μm in diameter and expressing on their surface β1-integrin proteins, contain mitochondria and lipid droplets together with ATP. We documented vesicle content with fluorescent-specific dyes and with the immunocytochemistry technique we confirmed that mitochondria and lipid droplets were co-localized in the same vesicle. Scanning electron microscopy and transmission electron microscopy confirmed that astrocytes shed from surface membrane vesicles of the same size as the ones detected by fluorescence microscopy. Our results report for the first time that cultured astrocytes, activated by repetitive stimulation of ATP released from neighboring cells, shed from their surface large membrane vesicles containing mitochondria and lipid droplets.

Ordered mesoporous materials (OMMs) are interesting matrixes for nanomedicine applications such as innovative drug delivery systems. Here, we compare the behavior of the widely studied SBA-15 mesoporous silica with that of the less investigated MSE (a periodic mesoporous organosilica whose silicon atoms are alternatively connected by means of -Si-O-Si- and -Si-CH(2)-CH(2)-Si- groups) toward the adsorption (pH 7.0 and 9.6) and in vitro release (pH 7.4; T = 37 degrees C) of an antimicrobial protein (hen egg white lysozyme). Both OMMs have a hexagonal ordered mesoporous structure and texture, as confirmed by SAXS, TEM, and N(2) adsorption isotherms, but differ for the chemical composition and surface charge density, as determined by ATR-FTIR spectroscopy and potentiometric titrations, respectively. Rather than the structural and textural features, the different chemical composition of SBA-15 and MSE seems to be responsible for the different lysozyme loading and release and for the different stability toward the lixiviating action of the physiological medium (pH 7.4; T = 37 degrees C)

The utility of 5-fluoro-5-deoxyribose (FDR) as an efficient bioconjugation agent for radiolabelling of the RGD peptides c(RGDfK) and c(RGDfC) is demonstrated. The bioconjugation is significantly superior to that achieved with 2-fluoro-2-deoxyglucose (FDG) and benefits from the location of the fluorine at C-5, and that ribose is a 5-membered ring sugar rather than a 6-membered ring. Both features favour ring opening to the aldehydic form of the sugar to promote smooth oxime ligation with aminooxy ether functionalised peptides. [(18)F]FDR was prepared in this study by synthesis from fluoride-18 using an automated synthesis protocol adapting that used routinely for [(18)F]FDG. c(RGDfK) was functionalised with an aminooxyacetyl group (Aoa) via its lysine terminus, while c(RGDfC) was functionalised with an aminooxyhexylmaleimide (Ahm) through a cysteine-maleimide conjugation. Bioconjugation of [(18)F]FDR to c(RGDfC)-Ahm proved to be more efficient than c(RGDfK)-Aoa (92% versus 65%). The unlabelled ((19)F) bioconjugates c(RGDfK)-Aoa-FDR and c(RGDfC)-Ahm-FDR were prepared and their in vitro affinity to purified integrin αvβ3 was determined. c(RGDfK)-Aoa-FDR showed the greater affinity. Purified "hot" bioconjugates c(RGDfK)-Aoa-[(18)F]FDR and c(RGDfC)-Ahm-[(18)F]FDR were assayed by incubation with MCF7, LNCaP and PC3 cell lines. In both cases the conjugated RGD peptides showed selectivity for PC3 cells, which express αvβ3 integrin, with the c(RGDfK)-Aoa-[(18)F]FDR demonstrating better binding, consistent with its higher in vitro affinity. The study demonstrates that [(18)F]FDR is an efficient bioconjugation ligand for RGD bioactive peptides.

TEM images of human lysozyme loaded on SBA-15 mesoporous silica were obtained through the immunogold staining (IGS) method. IGS is based on the specific interaction between proteins and colloidal gold-conjugated antibodies. Clear evidence that protein molecules are adsorbed both on the external and on the inner pore surface is presented.

From 1957 to 1980 in Sardinia, 182 cases of ALS with a mean annual incidence of 0.51 per 100,000 inhabitants and a prevalence rate of 3.65 per 100,000 inhabitants (prevalence day 21.10.1971) were observed. The disease was found to be more common in males, in subjects aged 50 to 70 years and in farmers and shepherds. Incidence in various areas of the island was found to be different. The common form was more frequent, had earlier onset and greater median survival rate.

Abstract. The water resources and hydrologic extremes in Mediterranean basins are heavily influenced by climate variability. Modeling these watersheds is difficult due to the complex nature of the hydrologic response as well as the sparseness of hydrometeorological observations. In this work, we present a strategy to calibrate a distributed hydrologic model, known as TIN-based Real-time Integrated Basin Simulator (tRIBS), in the Rio Mannu basin (RMB), a medium-sized watershed (472.5 km 2 ) located in an agricultural area in Sardinia, Italy. In the RMB, precipitation, streamflow and meteorological data were collected within different historical periods and at diverse temporal resolutions. We designed two statistical tools for downscaling precipitation and potential evapotranspiration data to create the hourly, high-resolution forcing for the hydrologic model from daily records. Despite the presence of several sources of uncertainty in the observations and model parameterization, the use of the disaggregated forcing led to good calibration and validation performances for the tRIBS model, when daily discharge observations were available. The methodology proposed here can be also used to disaggregate outputs of climate models and conduct high-resolution hydrologic simulations with the goal of quantifying the impacts of climate change on water resources and the frequency of hydrologic extremes within medium-sized basins.

In recent years, evidence is growing on the role played by gestational factors in shaping brain development and on the influence of intrauterine experiences on later development of neurodegenerative diseases including Parkinson's (PD) and Alzheimer's disease (AD). The nine months of intrauterine development and the first three years of postnatal life are appearing to be extremely critical for making connections among neurons and among neuronal and glial cells that will shape a lifetime of experience. Here, the multiple epigenetic factors acting during gestation - including maternal diet, malnutrition, stress, hypertension, maternal diabetes, fetal hypoxia, prematurity, low birth weight, prenatal infection, intrauterine growth restriction, drugs administered to the mother or to the baby - are reported, and their ability to modulate brain development, resulting in interindividual variability in the total neuronal and glial burden at birth is discussed. Data from recent literature suggest that prevention of neurodegeneration should be identified as the one method to halt the diffusion of neurodegenerative diseases. The "two hits" hypothesis, first introduced for PD and successfully applied to AD and other neurodegenerative human pathologies, should focus our attention on a peculiar period of our life: the intrauterine and perinatal periods. The first hit to our nervous system occurs early in life, determining a PD or AD imprinting to our brain that will condition our resistance or, alternatively, our susceptibility to develop a neurodegenerative disease later in life. In conclusion, how early life events contribute to late-life development of adult neurodegenerative diseases, including PD and AD, is emerging as a new fascinating research focus. This assumption implies that research on prevention of neurodegenerative diseases should center on events taking place early in life, during gestation and in the perinatal periods, thus presenting a new challenge to perinatologists: the prevention of neurodegenerative human diseases.