Design, synthesis and biological evaluation of pyrazolo-pyrimidines and related isosteres as inhibitors of protein kinases, potential antineoplastic agents

Design, synthesis and biological evaluation of pyrazolo-pyrimidines and related isosteres as inhibitors of protein kinases, potential antineoplastic agent

Metagenomic analysis of bacterial assemblages from Sardinian soils

The soil microbiota is investigated by technical approaches that are usually grouped in two main categories i) cultivation-based and ii) cultivation independent techniques. Only 0,1 to 1% of known soil bacteria species are today “culturable”. Most of soil species are not tractable using conventional cultivation methods and therefore these must be studied by cultivation independent techniques. The taxonomic composition of the bacterial fractions of soil has been recently investigated by employing the so called “omic” approaches. Deep sequencing of 16S rRNA based libraries have demonstrated an enormous diversities of bacteria inhabiting the soils. In the first chapter of this work we applied pyrosequencing of 16S rRNA libraries to analyze richness and diversity of bacterial communities of Sardinian soils managed with different grazing intensity. Our data indicated that grazing is associated to a dramatic shift of community structure with Firmicutes being positively associated to grazing intensity and becoming the dominant phylum in intensively grazed soils. No significant shifts were noted in the relative abundance at class, order or family taxonomic levels suggesting no drastic changes within phyla. Correlation analysis of bacterial communities composition and physical-chemical properties of soils highlighted significant associations between soil pH or C/N ratio and Firmicutes and Verrucomicrobia relative abundance. In second chapter of the present work we analyzed the effects of organic and mineral fertilization on soil microbiota communities. The organic fertilization source was cattle manure and slurry, while the mineral was represented by ENTEC. The experimental setup was designed to represent a typical land management system of a nitrogen vulnerable zone of central Sadinia. Our results indicated that systems receiving manure are characterized by a bacteria community enriched for species able to degrade complex organic compounds. On the contrary, systems receiving mineral fertilization were enriched for oligotrophic species adapter to nutrients limited environments

An integrated top-down and bottom-up proteomic platform to reveal potential salivary biomarkers of the rare disorders SAPHO syndrome, Wilson’s disease and Hereditary angioedema

Wilson’s disease, SAPHO syndrome and Hereditary angioedema are three rare disorders characterized by a wide spectrum of different clinical manifestations, which involve several organs and apparatus, making the diagnosis extremely difficult. In this study, the salivary proteome and peptidome of subjects affected by these pathologies has been investigated using mass spectrometry, through an integrated top-down and bottom-up platform, and compared with groups of healthy controls, with the aim to assess whether qualitative and quantitative variations of salivary proteins and peptides could be associated to the immune derangement distinctive of each disease and in order to have suggestions on potential specific salivary biomarkers. The analysis of the salivary proteome from patients affected by Wilson’s disease allowed to characterize new oxidized proteoforms of S100A8 and S100A9 and two fragments of the polymeric immunoglobulin receptor named ASVD and AVAD. Higher levels of these proteins and peptides observed in the patients are most likely connected to the oxidative stress, the activation of the inflammatory processes, and the hepatic damage caused by the altered copper transport and its subsequent accumulation in the organism, which is at the origin of the pathology. The observed increase of the level of α-defensins 2 and 4 may give a contribution to the development of the disease by the improvement of the free copper. The proteome of patients affected by SAPHO syndrome revealed a significant decrease of cystatins, histatins, and aPRPs, which are involved in the protection against infections, suggesting a reduced ability of these subjects to contrast bacteria colonization, in particular P. acnes which is a possible trigger of this disease. In particular, the lower levels of histatins and the higher frequency of S100A12 observed in patients with respect to controls, may be connected with the dysregulation of the innate immunity and the neutrophil response typical of SAPHO syndrome. Cystatin SN abundance decrease correlated with the disease duration, suggesting its reduced production during the chronic phase of the ~ 5 ~ disease, while histatins showed positive correlation with serum levels of the C reactive protein. In saliva of Hereditary angioedema patients, the increased percentage of peptides generated by the proteolytic cleavage by metalloproteinases indicates the intense metalloproteinase activity possibly connected to the activation of inflammatory pathways. Interestingly, in consideration of the possible role of cystatin B in enhancing the production of nitric oxide, and the higher salivary levels measured in the patients, we suggest that cystatin B may give a contribution to the vasodilatation and the vasopermeability responsible for the oedema formation, which is the main feature of this pathology. In conclusion, the results obtained in these studies clearly highlighted that the salivary proteome showed some features specific of the three pathologies. Even though these results have been obtained in a small cohort of patients, due to the difficult recruitment of subjects affected by rare disorders, and need further validation by using orthogonal techniques, they strongly suggest that saliva, with easy and non-invasive collection characteristics, could be a biofluid suitable for diagnostic applications

Optimization of a Loop-Mediated Isothermal Amplification Assay for On-Site Detection of Fusarium fujikuroi in Rice Seed

Fusarium fujikuroi, causing bakanae disease, is one of the most important seedborne pathogens of rice, the detection of which is paramount for seed certification and for preventing field infections. Molecular tests—qPCR and loop-mediated isothermal amplification (LAMP)—are replacing the blotter test in seed health procedures, due to higher sensitivity, specificity, fast turnaround results delivery, on-site application and the possibility of quantifying endophytic seed infections. A LAMP test, which had been previously developed with primers designed to target the elongation factor 1-α sequence of F. fujikuroi, was validated according to the international validation standard (EPPO, PM7/98) on thirty-four rice seed lines of different levels of susceptibility to the disease, thus comparing it to the blotter test and with two different DNA extraction procedures. The use of crude extracted DNA provided more sensitive results than the DNA extracted with the commercial kit Omega E.Z.N.A® Plant DNA kit. The results showed that the endophytic infection of F. fujikuroi is essential for the development of the disease in the field and that the minimum amount of the pathogen necessary for the development of the disease corresponds to 4.17 × 104 cells/µL. This study confirms the applicability of the LAMP technique on-site on rice seeds with fast and quantitative detection of the pathogen

Novel Perspectives on p53 Function in Neural Stem Cells and Brain Tumors

Malignant glioma is the most common brain tumor in adults and is associated with a very poor prognosis. Mutations in the p53 tumor suppressor gene are frequently detected in gliomas. p53 is well-known for its ability to induce cell cycle arrest, apoptosis, senescence, or differentiation following cellular stress. That the guardian of the genome also controls stem cell self-renewal and suppresses pluripotency adds a novel level of complexity to p53. Exactly how p53 works in order to prevent malignant transformation of cells in the central nervous system remains unclear, and despite being one of the most studied proteins, there is a need to acquire further knowledge about p53 in neural stem cells. Importantly, the characterization of glioma cells with stem-like properties, also known as brain tumor stem cells, has opened up for the development of novel targeted therapies. Here, we give an overview of what is currently known about p53 in brain tumors and neural stem cells. Specifically, we review the literature regarding transformation of adult neural stem cells and, we discuss how the loss of p53 and deregulation of growth factor signaling pathways, such as increased PDGF signaling, lead to brain tumor development. Reactivation of p53 in brain tumor stem cell populations in combination with current treatments for glioma should be further explored and may become a viable future therapeutic approach

Are nonsingular black holes with super-Planckian hair ruled out by S2 star data?

We propose a novel nonsingular black-hole spacetime representing a strong deformation of the Schwarzschild solution with mass $M$ by an additional hair $\ell$, which may be hierarchically larger than the Planck scale. Our black-hole model presents a de Sitter core and $\mathcal{O}(\ell^2/r^2)$ slow-decaying corrections to the Schwarzschild solution. Our black-hole solutions are thermodynamically preferred when $0.2 \lesssim \ell/GM \lesssim \, 0.3$ and are characterized by strong deviations in the orbits of test particles from the Schwarzschild case. In particular, we find corrections to the perihelion precession angle scaling linearly with $\ell$. We test our model using the available data for the orbits of the S2 star around $\text{SgrA}^*$. These data strongly constrain the value of the hair $\ell$, casting an upper bound on it of $\sim \, 0.47 \, GM$, but do not rule out the possible existence of regular black holes with super-Planckian hair.Comment: 6 pages, 2 figures, 1 table. Revised version: Addendum and some references have been adde

Preliminary finding of a randomized, double-blind, placebo-controlled, crossover study to evaluate the safety and efficacy of 5-hydroxytryptophan on REM sleep behavior disorder in Parkinson's disease

Altered serotonergic neurotransmission may contribute to the non-motor features commonly associated with Parkinson's disease (PD) such as sleep disorders. The 5-hydroxytryptophan (5-HTP) is the intermediate metabolite of L-tryptophan in the production of serotonin and melatonin. The purpose of this study was to compare the effects of 5-HTP to placebo on REM sleep behavior disorder (RBD) status in patients with PD

Terrestrial Laser Scanner and Close Range Photogrammetry point clouds accuracy assessment for the structure deformations monitoring

In this paper we show the results of several tests carried out using methods and instrumentation typical of an architectural survey, along with a set of metrological instrumentation, on a Reinforced Concrete (RC) beam subjected to increasing loads. The goal was to assess the accuracy in the displacements estimated by a medium quality terrestrial laser scanner (TLS) Focus 3d from Faro Technologies, and the low-cost digital camera Canon PowerShot S110 used in a Close Range Photogrammetry (CRP) survey. The software used for scan-data and point clouds processing was Reconstructor JRC Software v. 3.1.0, maintained by Gexcel Ltd, while the images processing was performed with the software Photoscan from Agisoft, which implements Structure from Motion (SfM) approach. Two processing strategies were used in the point clouds comparison: mesh2mesh and modelling the beam behavior fitting the contours of the beam with second order polynomials. Comparisons between the TLS and CRP techniques and the metrological equipment used in parallel highlighted the limits and potentialities of the two geomatic techniques used. It has been shown that modeling the behavior of the beam leads to significantly better results than using the mesh2mesh comparison. For the CRP the increase in accuracy was in the order of 40%, while for the TLS of 50%

GIANO, the high resolution IR spectrograph of the TNG: geometry of the echellogram and strategies for the 2D reduction of the spectra

GIANO is the IR high resolution spectrograph of the TNG. It covers the 950-2450 nm wavelengths range in a single shot at a resolving power of R=50,000. This document describes the first fundamental steps of the data reduction, namely eliminating the curvature of the traces and the tilt of the slit images. These effects can be accurately modeled and corrected using a physical model of the instrument. We find that the curvature and tilt parameters did not vary during the whole lifetime of the instrument. In particular, they were not affected by thermal cycles or by the works performed to mount the spectrometer on its new interface. A similar ab-initio modeling is also applied to the wavelength calibration that can be accurately (0.03 pixel r.m.s.) defined using a minimum number of parameters to fit. This approach is particularly useful when using a calibration source with an irregular wavelengths coverage; e.g. for the U-Ne lamp that has only few lines in the 2000 nm - 2300 nm wavelengths range