Search

Weakening the Spindle Assembly Checkpoint by reduced expression of its components induces chromosome instability and aneuploidy that are hallmarks of cancer cells. The tumor suppressor p14ARF is overexpressed in response to oncogenic stimuli to stabilize p53 halting cell progression. Previously, we found that lack or reduced expression of p14ARF is involved in the maintenance of aneuploid cells in primary human cells, suggesting that it could be part of a pathway controlling their proliferation. To investigate this aspect further, p14ARF was ectopically expressed in HCT116 cells after depletion of the Spindle Assembly Checkpoint MAD2 protein that was used as a trigger for aneuploidy. p14ARF Re-expression reduced the number of aneuploid cells in MAD2 post-transcriptionally silenced cells. Also aberrant mitoses, frequently displayed in MAD2-depleted cells, were decreased when p14ARF was expressed at the same time. In addition, p14ARF ectopic expression in MAD2-depleted cells induced apoptosis associated with increased p53 protein levels. Conversely, p14ARF ectopic expression did not induce apoptosis in HCT116 p53KO cells. Collectively, our results suggest that the tumor suppressor p14ARF may have an important role in counteracting proliferation of aneuploid cells by activating p53-dependent apoptosis. J. Cell. Physiol. 231: 336–344, 2016. © 2015 Wiley Periodicals, Inc.

Understanding sandbar dynamics and variability is integral to developing a predictive capacity for nearshore flows, sediment transport, morphological change, and ultimately for determining coastline exposure to damaging storm waves. Here we report on a nearshore bathymetric data set from the U.S. Pacific Northwest (PNW) that stretches from Point Grenville, Washington to Cascade Head, Oregon, over approximately 260 km in the alongshore and includes 8 distinct littoral cells. We describe and quantify the morphological variability of sandbars on a regional scale, using 560 individual cross-shore transects, as well as attempt to explain the inter-littoral cell variability via relationships to various environmental parameters. The cross-shore extent of the bar zone extends over 1 km from the shoreline in the northern part of the study area, but only to about 600 m from the shoreline in the southern part. Maximum bar crest depths are typically 7 m below MLLW. Bar heights range from a step in the cross-shore profile to over 3 m from crest to trough. The northernmost littoral cells typically have two or more subtidal sandbars per cross-shore profile whereas the littoral cells in the southern part of our study area have only one bar. The mean depths of the bars, however, are much more consistent across littoral cells even while the upper shoreface slope significantly increases from north to south, requiring that the maximum bar distance from the shoreline decreases from north to south. Results from a limited study of the temporal variability suggest that while data collected over large spatial scales captures significant amounts of overall sandbar variability, it does not completely characterize the variability over the entirety of the net offshore migration cycle.

High sediment contamination in the coastal area of Priolo Bay, adjacent to the highly-polluted Augusta Harbour, poses serious risks for the benthic communities inhabiting the area. Nevertheless, the transfer of trace elements and consequent bioaccumulation in the biota is an overlooked issue. This study aimed to assess the transfer and bioaccumulation patterns of As, Cd, Ni and Hg to the dominant macroalgae and benthic invertebrates of Priolo Bay. Results revealed different patterns among trace elements (TEs), not driven by sediment contamination but rather by element-specific behaviour coupled with sediment physicochemical properties. Specifically, As accumulated in macroalgae but not in invertebrates, indicating bioavailability of dissolved As only, and a lack of effective trophic transfer. Ni was confined to surface sediment and transfer to biota was not highlighted. Cd and Hg showed the highest concentrations in invertebrates and bioaccumulated especially in filter feeders and carnivores, revealing the importance of suspended particulate and diet as transfer pathways. Total organic carbon (TOC), fine-grained sediments and redox potential were the most important sediment features in shaping the sediment contamination spatial patterns as well as those of TE transfer and bioaccumulation. In particular, As and Cd transfer to macroalgae, and especially Hg bioaccumulation in benthic invertebrates was controlled by sediment properties, resulting in limited transfer and accumulation in the most contaminated stations.

The aim of this study was to obtain information on the presence and levels of hazardous organic pollutants in groundwater located close to solid waste landfills. Eighty-two environmental contaminants, including 16 polycyclic aromatic hydrocarbons (PAHs), 20 volatile organic compounds (VOCs), 29 polychlorinated biphenyls (PCBs), 7 dioxins (polychlorinated dibenzo-p-dioxins, PCDDs) and 10 furans (polychlorinated dibenzofurans, PCDFs) were monitored in areas characterised by different geological environments surrounding three municipal solid waste landfills (Palermo, Siculiana and Ragusa) in Sicily (Italy) in three sampling campaigns. The total concentrations of the 16 PAHs were always below the legal threshold. Overall, the Fl/Fl + Py diagnostic ratio revealed that PAHs had a petrogenic origin. VOC levels, except for two notable exceptions near Palermo landfill, were always below the legal limit. As concerns PCB levels, several samples were found positive with levels exceeding the legal limits. It is worth noting that the % PCB distribution differs from that of commercial compositions. In parallel, some samples of groundwater containing PCDDs and PCDFs exceeding the legal threshold were also found. Among the 17 congeners monitored, the most abundant were the highest molecular weight ones.

Debris-flows runout estimations are of crucial importance to delineate the potentially hazardous areas and to make reliable assessment of the level of risk of urban areas. Especially in recent years, several researches have been conducted in order to define predicitive models. But, existing runout estimation methods need input parameters that can be difficult to estimate. Recent experimental researches have also allowed the assessment of the physics of the debris flows. But, the major part of the experimental studies analyze the basic kinematic conditions which determine the phenomenon evolution. Experimental program has been recently conducted at the Hydraulic laboratory of the Department of Civil, Environmental, Aerospatial and of Materials (DICAM) – University of Palermo (Italy). The experiments, carried out in a laboratory flume appositely constructed, were planned in order to evaluate both the effect of sediment concentration and the influence of the geometrical parameters (such as the slope and the geometrical characteristics of the confluences to the main channel) on the propagation phenomenon of the debris flow and its deposition. The laboratory flume reproduces the protection channel which is under construction in Giampilieri (Messina – Italy). The experimental apparatus includes a high-precision camera allowing the estimation of hyperconcentrated flow velocity by applying the VIA technique. Attention is particularly focused on flowing of sediment-water mixture for different geometrical characteristics (slope, angle,..) of the inflow channels and/or by introducing roughness elements on the bed. The propagation phenomenon is analyzed and discussed for different concentrations of sediments

Posidonia oceanica mat is considered a long-term bioindicator of contamination. Storage and sequestration of trace elements and organic carbon (Corg) were assessed in dead P. oceanica mat and bare sediments from a highly polluted coastal marine area (Augusta Bay, central Mediterranean). Sediment elemental composition and sources of organic matter have been altered since the 1950s. Dead P. oceanica mat displayed a greater ability to bury and store trace elements and Corg than nearby bare sediments, acting as a long-term contaminant sink over the past 120 yr. Trace elements, probably associated with the mineral fraction, were stabilized and trapped despite die-off of the overlying P. oceanica meadow. Mat deposits registered historic contamination phases well, confirming their role as natural archives for recording trace element trends in marine coastal environments. This sediment typology is enriched with seagrass-derived refractory organic matter, which acts mainly as a diluent of trace elements. Bare sediments showed evidence of inwash of contaminated sediments via reworking; more rapid and irregular sediment accumulation; and, because of the high proportions of labile organic matter, a greater capacity to store trace elements. Through different processes, both sediment typologies represent a repository for chemicals and may pose a risk to the marine ecosystem as a secondary source of contaminants in the case of sediment dredging or erosion.

Optimization of a new lead promoting the readthrough of the nonsense mutations for CFTR rescue in human CF cells Laura Lentini, Raffaella Melfi, Sara Baldassano, Marco Tutone, Aldo Di Leonardo, Andrea Pace, Ivana Pibiri Department of Biological, Chemical and Pharmaceutical Sciences and Technologies (STEBICEF), University of Palermo Background and rationale Cystic Fibrosis patients with nonsense mutations in the CFTR gene have a more severe form of the disease. Nonsense mutations represent about 10% of the mutations that affect the CFTR gene and they are frequently associated to the classical F508 mutation (1). A potential treatment for this genetic alteration is to promote the translational readthrough of premature termination codons (PTCs) by Translational Read-Through-Inducing Drugs (TRIDs) (2-4). Hypothesis and objectives Our objective is to evaluate the functionality of the CFTR channel after treatment with a new molecule that we individuated in a precedent FFC project, and the activity of new lead molecules in cells stably expressing a nonsense-CFTR-mRNA (ns CFTR) in CF cellular model systems. We want also to study the supramolecular interactions among TRIDs, CFTR mRNA and the ribosomal A-site to identify the biological target and the mechanism of action. Essential methods QSAR, carried out on the basis of our preliminary results, will allow to achieve lead optimization and synthesize then a small library of analogs to be tested and compared to the Lead. We will mutagenize the CFTR cDNA by introducing the most diffuse nonsense mutations. Subsequently, FRT cells engineered with the vector expressing mutagenized nsCFTR, and nonsense-CF-human broncoepithelial cells will be grown in the air-liquid culture system to reproduce in vitro the epithelial organization. CFTR expression after treatments with our molecule will be evaluated by biomolecular techniques. CFTR activity will be revealed by specific CFTR-functionality assays. Finally, in vitro-in vivo (Zebrafish model) analyses of the safety profile for the set of synthesized molecules will complete the study. Preliminary results We screened the activity of several molecules synthetized by us in a precedent FFC project, identifying some molecules that showed high readthrough activity associated to the expression of the CFTR protein in ns CF immortalized cells. Expected final results and their significance We are confident that our findings will provide the validation of molecules with readthrough activity for the recovery of the CFTR function. Moreover, our pre-clinical study will assess the presence of toxic effects caused by the molecules in vivo. References 1. Sermet-Gaudelus I, Boeck KD, Casimir GJ, Vermeulen F, Leal T, Mogenet A, Roussel D, Fritsch J, Hanssens L, Hirawat S, Miller NL, Constantine S, Reha A, Ajayi T, Elfring GL, Miller LL. Ataluren (PTC124) induces cystic fibrosis transmembrane conductance regulator protein expression and activity in children with nonsense mutation cystic fibrosis., Am J RespirCrit Care Med. 2010 Nov 15;182(10):1262-72. 2. Lentini L, Melfi R, Di Leonardo A, Spinello A, Barone G, Pace A, Palumbo Piccionello A, Pibiri I. Towards a rationale for the PTC124 (Ataluren) promoted read-through of premature stop codons: a computational approach and GFP-reporter cell-based assay. Mol. Pharm. 2014 11, 653-664. 3. Pibiri I, Lentini L, Melfi R, Gallucci G, Pace A, Spinello A, Barone G, Di Leonardo A. Enhancement of premature stop codon readthrough in the CFTR gene by Ataluren (PTC124) derivatives European Journal of Medicinal Chemistry 06/2015; 101. 4. Nagel-Wolfrum K, Möller F, Penner I, Baasov T4, Wolfrum U. Targeting Nonsense Mutations in Diseases with Translational Read-Through-Inducing Drugs (TRIDs), BioDrugs. 2016 Apr;30(2):49-74.

The aim of the study was to identify which groups of women contribute to interinstitutional variation of caesarean delivery (CD) rates and which are the reasons for this variation. In this regard, 15,726 deliveries from 11 regional centers were evaluated using the 10-group classification system. Standardized indications for CD in each group were used. Spearman’s correlation coefficient was used to calculate (1) relationship between institutional CD rates and relative sizes/CD rates in each of the ten groups/centers; (2) correlation between institutional CD rates and indications for CD in each of the ten groups/centers. Overall CD rates correlated with both CD rates in spontaneous and induced labouring nulliparous women with a single cephalic pregnancy at term (P=0.005). Variation of CD rates was also dependent on relative size and CD rates in multiparous women with previous CD, single cephalic pregnancy at term (P<0.001). As for the indications, “cardiotocographic anomalies” and “failure to progress” in the group of nulliparous women in spontaneous labour and “one previous CD” in multiparous women previous CD correlated significantly with institutional CD rates (P=0.021,P=0.005, andP<0.001, resp.). These results supported the conclusion that only selected indications in specific obstetric groups accounted for interinstitutional variation of CD rates.

The City on the Hill has always willfully confused race with righteousness. Since the Puritans' arrival, dominant American political narratives have emphasized the differential moral worth of racial/ethnic populations: whites versus Native Americans, Yankees versus Irish, planters versus slaves, native-born whites versus "the refuse of Europe", and, on the West Coast, whites versus Mexicans and Asians. This Puritanical inheritance has been transformed, however, in the climate of center-right convergence in national politics and in the process has lost its overtly racist character. The white ethnic community construct arose from an extraordinarily complex historical ground, and this complexity was reflected in its multiple expressions and political uses. "Women's culture", a locution in increasing use over the 1970s, evokes both the American feminist shift in emphasis from male-female to female-female relations and what historian Alice Echols has labeled the decline from radical to cultural feminism over the course of the late 1960s to early 1970s.

Many motile microorganisms react to environmental light cues with a variety of motility responses guiding cells towards better conditions for survival and growth. The use of spatial light modulators could help to elucidate the mechanisms of photo-movements while, at the same time, providing an efficient strategy to achieve spatial and temporal control of cell concentration. Here we demonstrate that millions of bacteria, genetically modified to swim smoothly with a light controllable speed, can be arranged into complex and reconfigurable density patterns using a digital light projector. We show that a homogeneous sea of freely swimming bacteria can be made to morph between complex shapes. We model non-local effects arising from memory in light response and show how these can be mitigated by a feedback control strategy resulting in the detailed reproduction of grayscale density images., 8 pages, 6 figures, eLife 2018 (accepted)

The spindle assembly checkpoint (SAC) is a cellular surveillance mechanism that ensures the fidelity of chromosomes segregation. Reduced expression of some of its components weakens the SAC and induces chromosome instability and aneuploidy, which are both well-known hallmarks of cancer cells. Centromere protein-E (CENP-E) is a crucial component of the SAC and its function is to facilitate kinetochore microtubule attachment required to achieve and maintain chromosome alignment. The present study investigates the possible role of p14

Granulometry, mineralogy, and trace element concentrations are determined in marine sediments from thirty-six sampling sites in the littoral environment of the Gulf of Milazzo (NE Sicily). Sediment samples were collected in August 2008, along 18 seaward transects, at water depths of ‐10, ‐20 and ‐30 m, by using a Van Veen grab. Grain-size analysis shows predominance of sand (56%) and silt (35%) fractions with respect to clay (7%) and gravel (2%) fractions. Bulk mineralogical analysis documents the presence of quartz, micas, feldspars, calcite, and chlorite, which reflect erosion processes affecting the Kabilian-Calabrian Units. Concentrations of most trace elements in the deeper sediments were notably higher than shallower ones, due to the gradual increase of the fine fraction (

We present a stochastic description of a model of N mutually repelling active spheres in the presence of external fields and characterize its steady state behavior. To reproduce the effects of the experimentally observed persistence of the trajectories of the active particles we consider a Gaussian forcing having a non vanishing correlation time $\tau$, whose finiteness is a measure of the activity of the system. With these ingredients we show that it is possible to develop a statistical mechanical approach similar to the one employed in the study of equilibrium liquids and to obtain the explicit form of the many-particle distribution function by means of the multidimensional unified colored noise approximation. Such a distribution plays a role analogous to the Gibbs distribution in equilibrium statistical mechanics and provides a complete information about the microscopic state of the system. From here we develop a method to determine the one and two-particle distribution functions in the spirit of the Born-Green-Yvon (BGY) equations of equilibrium statistical mechanics. The resulting equations which contain extra-correlations induced by the activity allow to determine the stationary density profiles in the presence of external fields, the pair correlations and the pressure of active fluids. In the low density regime we obtain the effective pair potential $\phi(r)$ acting between two isolated particles separated by a distance, $r$, showing the existence an effective attraction between them induced by activity. Based on these results, in the second half of the paper we propose a mean field theory as an approach simpler than the BGY hierarchy and use it to derive a van der Waals expression of the equation of state, which can serve as the basis to understand the phase behavior of active fluids., Comment: 18 pages, 4 figures, Soft Matter, September 11, 2015

Integration of active matter in larger micro-devices can provide an embedded source of propulsion and lead to self-actuated micromachining systems that do not rely on any external power or control apparatus. Here we demonstrate that Janus colloids can self-assemble around micro-fabricated rotors in reproducible configurations with a high degree of spatial and orientational order. The final configuration maximizes the torque applied on the rotor leading to a unidirectional and steady rotating motion. We discuss how the interplay between geometry and dynamical behavior consistently leads to the self-assembly of autonomous micromotors starting from randomly distributed building blocks., 17 pages, 6 figures

Clinical data suggest that beyond-progression, the blockade of angiogenesis is associated with improved survivals in colorectal cancer. We conducted a systematic review to investigate the therapeutic effects of antiangiogenic drugs administered as later lines of treatment in patients already progressed to a previous anti-VEGF based treatment. An extensive literature search was conducted. Hazard ratios (HR) for progression (PFS) and death (OS) were extracted. An inverse-variance meta-analysis model was implemented. 6 randomized controlled trials were retrieved, including 3407 patients, treated with different antiangiogenic drugs. All of them had progressed during or after a previous line of treatment with bevacizumab. Overall, both PFS (HR=0.63, P P < 0.001) were significantly increased with the use of antiangiogenic drug. No heterogeneity was observed despite different drugs. Protracted inhibition of the VEGF pathway is associated with a significant improvement of both PFS and OS, independently from the antiangiogenic agent used.

Most human tumors are characterized by aneuploidy that is believed to be the consequence of chromosomal instability (CIN). The mechanism(s) leading to aneuploidy and the pathways that allow its tolerance are not completely understood. The Spindle Assembly Checkpoint (SAC) is a cellular surveillance mechanism working during mitosis, and alterations of genes that encode components of the SAC weakening the mitotic checkpoint, induce aneuploidy by chromosome mis-segregation. We induced aneuploidy in near-diploid tumor cells by simultaneous depletion of the SAC proteins MAD2 and BUBR1 by RNA interference in the attempt to gain further insight on the cellular responses to aneuploidy. Individual reduction of MAD2 and BUBR1 protein levels caused defective mitosis and aneuploidy, while co-depletion of MAD2 and BUBR1 caused cell cycle arrest and cell death in addition to aneuploidy. The simultaneous reduction of the two SAC proteins induced high percentage of hyperdiploid cells and p53 stabilization suggesting that hyperdiploidy could activate a p53 controlled pathway. The results indicate that p53 is required to induce cell cycle arrest and cell death when the mitotic checkpoint is strongly perturbed, thereby preventing aneuploid cell propagation.

Aurora-A is a centrosome-associated serine/threonine kinase that is overexpressed in multiple types of human tumors. Primarily, Aurora-A functions in centrosome maturation and mitotic spindle assembly. Overexpression of Aurora-A induces centrosome amplification and G 2 /M cell cycle progression. Recently, it was observed that overexpression of Aurora-A renders cells resistant to cisplatin (CDDP)-, etoposide-, and paclitaxel-induced apoptosis.Our results indicate that already in initial stages of cancer progression Aurora-A overexpression could have a major role in inducing supernumerary centrosomes and aneuploidy, as shown by immunohistochemistry on tissue sections from various stages of human colon cancer. Aneuploidy was also observed after Aurora-A ectopic overexpression in colon cancer cells with MIN phenotype. Silencing of Aurora-A by RNA interference in tumor cell lines triggered arrest of the cell cycle associated to apoptosis/ mitotic catastrophe. Finally, Aurora-A transcriptional silencing seems to confer cancer cells a greater sensitivity to chemotherapy by vincristine, indicating Aurora-A as a possible gene target in cancer therapy.

In this work the sterilization with dense carbon dioxide of poly( l -lactic acid) (PLLA) porous scaffolds intended for tissue engineering applications was investigated with the main objective of confirming the three-dimensional efficacy of the treatment and of analysing the scaffold properties after CO2 treatment. For this purpose the scaffold was contaminated with a conventional bacterium (Escherichia coli) and with spores (Streptomyces coelicolor), a species more fascinating and difficult to inactivate. Contamination was performed in such a way to soak the whole matrix with bacteria and spores. The effect of pressure and treatment time on the efficacy of the sterilization was evaluated. The E. coli was eradicated from the whole matrix after just 5 min of exposure to supercritical CO2 at 10 MPa and 40 °C. The spores required more severe conditions due to their different and very resistant structures since complete inactivation was obtained after 360 min of exposure at 30 MPa and 40 °C or at 30 MPa and 30 °C after the addition of 200 ppm of H2O2 to near critical CO2. The treatment with dense CO2 did not alter the biocompatibility and the structure of the scaffold as demonstrated by biological culture tests and calorimetric and SEM analyses. Collected data suggest that dense CO2 is a promising alternative to conventional sterilization techniques to sterilize biodegradable PLLA scaffolds.

This paper investigates the utility and the efficiency of the digital image-technique for remote monitoring of surface velocity in hyper-concentrated flow by the aid of data collected during experiments conducted in the laboratory flume. In particular the present paper attention is focused on the estimation procedure of the velocity vectors and on their sensitivity with parameters (number of images, spatial resolution of interrogation area, ) of the images processing procedure.

Aneuploidy, the unbalanced number of chromosomes in a cell, is considered a prevalent form of genetic instability and is largely acknowledged as a condition implicated in tumorigenesis. Epigenetic alterations like DNA hypomethylation have been correlated with cancer initiation/progression. Furthermore, a growing body of evidence suggests the involvement of epigenome-wide disruption as a cause of global DNA hypomethylation in aneuploidy generation. Here, we report that the DNA hypomethylating drug 5-aza-2′-deoxycytidine (DAC), affects the correct ploidy of nearly diploid HCT-116 human cells by altering the methylation pattern of the chromosomes. Specifically, we show that a DAC-induced reduction of 5-Methyl Cytosine at the pericentromeric region of chromosomes correlates with aneuploidy and mitotic defects. Our results suggest that DNA hypomethylation leads to aneuploidy by altering the DNA methylation landscape at the centromere that is necessary to ensure proper chromosomes segregation by recruiting the proteins necessary to build up a functional kinetochore.

This paper describes the application of image-based technique for mapping surface velocity of hyper-concentrated flows. The analysis is conducted with the aid of data collected in a scale laboratory flume constructed at the Hydraulic laboratory of the Department of Civil, Environmental, Aerospatial and of Mate-rials Engineering (DICAM) – University of Palermo (Italy). A fully digital images-based technique has been applied to record a large amount of high resolution images identifying simultaneously the position of points in different time instants. The sensitivity analysis of the estimated flow velocity with the acquisition conditions and the number of processed frames is performed

Premature stop codons are the result of nonsense mutations occurring within the coding sequence of a gene. These mutations lead to the synthesis of a truncated protein and are responsible for several genetic diseases. A potential pharmacological approach to treat these diseases is to promote the translational readthrough of premature stop codons by small molecules aiming to restore the full-length protein. The compound PTC124 (Ataluren) was reported to promote the readthrough of the premature UGA stop codon, although its activity was questioned. The potential interaction of PTC124 with mutated mRNA was recently suggested by molecular dynamics (MD) studies highlighting the importance of H-bonding and stacking π-π interactions. To improve the readthrough activity we changed the fluorine number and position in the PTC124 fluoroaryl moiety. The readthrough ability of these PTC124 derivatives was tested in human cells harboring reporter plasmids with premature stop codons in H2BGFP and FLuc genes as well as in cystic fibrosis (CF) IB3.1 cells with a nonsense mutation. Maintaining low toxicity, three of these molecules showed higher efficacy than PTC124 in the readthrough of the UGA premature stop codon and in recovering the expression of the CFTR protein in IB3.1 cells from cystic fibrosis patient. Molecular dynamics simulations performed with mutated CFTR mRNA fragments and active or inactive derivatives are in agreement with the suggested interaction of PTC124 with mRNA.