HaRas activates the NADPH oxidase complex in human neuroblastoma cells via extracellular signal-regulated kinase 1/2 pathway

In this study we have investigated the effects of the small GTP-binding-protein Ras on the redox signalling of the human neuroblastoma cell line, SK-N-BE stably transfected with HaRas(Val12). The levels of reactive oxygen species (ROS) and superoxide anions were significantly higher in HaRas(Val12) expressing (SK-HaRas) cells than in control cells. The treatment of cells with 4-(2-aminoethyl) benzenesulfonylfluoride, a specific inhibitor of the membrane superoxide generating system NADPH oxidase, suppressed the rise in ROS and significantly reduced superoxide levels produced by SK-HaRas cells. Moreover, HaRas(Val12) induced the translocation of the cytosolic components of the NADPH oxidase complex p67(phox) and Rac to the plasma membrane. These effects depended on the mitogen-activated protein kinase kinase/extracellular signal-regulated kinase (MEK/ERK1/2) pathway, as the specific MEK inhibitor, PD98059, prevented HaRas-mediated increase in ROS and superoxide anions. In contrast, the specific phosphoinositide 3-kinase (PI3K) inhibitors LY294002 and wortmannin were unable to reverse the effects of HaRas(Val12). Moreover, cholinergic stimulation of neuroblastoma cells by carbachol, which activated endogenous Ras/ERK1/2, induced a significant increase in ROS levels and elicited membrane translocation of p67(phox) and Rac. ROS generation induced by carbachol required the activation of ERK1/2 and PI3K. Hence, these data indicate that HaRas-induced ERK1/2 signalling selectively activates NADPH oxidase system in neuroblastoma cells.     

PanDelos: a dictionary-based method for pan-genome content discovery

Background

Pan-genome approaches afford the discovery of homology relations in a set of genomes, by determining how some gene families are distributed among a given set of genomes. The retrieval of a complete gene distribution among a class of genomes is an NP-hard problem because computational costs increase with the number of analyzed genomes, in fact, all-against-all gene comparisons are required to completely solve the problem. In presence of phylogenetically distant genomes, due to the variability introduced in gene duplication and transmission, the task of recognizing homologous genes becomes even more difficult. A challenge on this field is that of designing fast and adaptive similarity measures in order to find a suitable pan-genome structure of homology relations.

Results

We present PanDelos, a stand alone tool for the discovery of pan-genome contents among phylogenetic distant genomes. The methodology is based on information theory and network analysis. It is parameter-free because thresholds are automatically deduced from the context. PanDelos avoids sequence alignment by introducing a measure based on k-mer multiplicity. The k-mer length is defined according to general arguments rather than empirical considerations. Homology candidate relations are integrated into a global network and groups of homologous genes are extracted by applying a community detection algorithm.

Conclusions

PanDelos outperforms existing approaches, Roary and EDGAR, in terms of running times and quality content discovery. Tests were run on collections of real genomes, previously used in analogous studies, and in synthetic benchmarks that represent fully trusted golden truth. The software is available at https://github.com/GiugnoLab/PanDelos.

     

Constitutive αVβ3 integrin-mediated adhesion of human lymphoid B cells to vitronectin substrate

Adherence to cells and matrices participates in lymphocyte migration and tissue localization and contributes to the regulation of growth and differentiation of the lymphoid cells. The adherence is mainly mediated by three families of cell-surface proteins: integrins, immunoglobulin (Ig)-related molecules, and selectins. Integrins recognize Ig-related molecules such as ICAMs as well as fibronectin (FN), vitronectin (VN), and other matrix proteins. In this study, the in vitro adhesive properties of two Epstein—Barr virus-carrying B lymphoblastoid cell lines, IB-4 and NAD-20, were compared. IB-4 cells grow as a monolayer in contrast to NAD-20 cells, which grow as cell clusters. IB-4 cells were found to adhere to the tissue culture vessel through a component of the fetal bovine serum. By using blocking monoclonal antibodies to cell-surface molecules and serum proteins, IB-4 cells were found to use αVβ3 integrin (CD51/CD61) and serum VN as the adhesive molecules. αVβ3 integrin also mediated adhesion of IB-4 cells to human serum VN and to purified VN and FN. This constitutive adherence was not enhanced by phorbol ester treatment and was inhibited by RGD-containing peptides, in contrast to the homotypic adhesion of NAD-20 cells, which was mediated by β2 integrin CD11a/CD18 and its ligand ICAM-1 (CD54). Since VN is a component of both lymphoid tissue matrix and plasma, adhesion to this protein may affect functions and activities of B lymphocytes.     

Effects of homogenization pressure on the survival and growth of some food spoilage and pathogenic micro-organisms

The effectiveness of pressures ranging from 150 to 2000 bar, applied by means of a continuous homogenizer, on the cell viability of spoilage or pathogenic micro-organisms, was evaluated. Moreover, the effects of microstructural modifications of food systems associated with the treatments on cell viability and on its subsequent evolution were investigated. Modulation of the homogenization pressure allowed strong instantaneous reductions of the initial numbers of Listeria monocytogenes, Yersinia enterocolitica, Bacillus subtilis and the yeast Yarrowia lipolytica. The combined effects of the initial pressure treatment and space reduction, resulting from the microstructural modifications of food systems, increased the safety and the shelf-life both of water in oil and oil in water emulsions.     

VARIATIONS OF PSP TOXIN PROFILES DURING DIFFERENT GROWTH PHASES IN GYMNODINIUM CATENATUM (DINOPHYCEAE) STRAINS ISOLATED FROM THREE LOCATIONS IN THE GULF OF CALIFORNIA,MEXICO1

In vitro experiments were performed with Gymnodinium catenatum Graham strains isolated from three locations in the Gulf of California to determine the variability in toxicity and toxin profiles. Strains were cultivated in GSe at 20°C±1°C, 150 μmol photons·m^?2·s^?1 (12:12 light:dark cycle), and harvested during different growth phases. Growth rates were higher than in previous studies, varying between 0.70 and 0.82 day^?1. The highest cell yields were reached at 16 and 19 days, with maximum densities between 1090 and 3393 cells·mL^?1. Bahía de La Paz (BAPAZ) and Bahía de Mazatlán (BAMAZ) were the most toxic (101 pg STXeq·cell^?1), whereas strains from Bahía Concepción (BACO) were the least toxic (13 pg STXeq·cell^?1). A strain isolated from cyst germination was one of the least toxic strains. No significant changes in toxin content with culture age were observed (0.2 and 0.6 pmol paralytic shellfish poisoning·cell^?1). All strains contained neosaxitoxin (NEOSTX), decarbamoyl‐saxitoxin (dcSTX), decarbamoyl‐gonyautoxin‐2,‐3, (dcGTX2‐3), N‐sulfo‐carbamoylsaxitoxin (B1), N‐sulfo‐carbamoylneosaxitoxin (B2), and N‐sulfo‐carbamoylgonyautoxin‐2,‐3 (C1‐2). Bahía Concepción strains had the highest content of C1; BAPAZ and BAMAZ strains had a higher percentage of NEOSTX. Differences in toxin composition with culture age were observed only in BAMAZ and BAPAZ strains. Cultures with a higher percentage of long chains had more NEOSTX, while those with a higher proportion of short chains had a lower content of NEOSTX. Gulf of California strains are characterized by a high proportion of NEOSTX, and seem to have evolved particular physiological responses to their environment that are reflected in the toxin profile, suggesting different populations.     

Reversible carnivory in Oxytricha bifaria: a peculiar ecological adaptation

The adaptive significance of the giant form of Oxytricha bifaria has been studied. It proved to be a carnivorous phase of a bacterivorous ciliate, rather than a cannibalistic one. By such a dramatic reversal in the feeding behavior O. bifaria, a primary consumer, can reversibly become a secondary one.     

Course of the degeneration of the primary visual system in albino quails with glaucoma

In the quail, a sex-linked albino mutation is associated with buphthalmic glaucoma. This progressive disorder is detectable 3 to 6 months after hatching. Its development leads to a degeneration of retinal projection according to a relatively orderly sequence progressing from the tegmentum to the tectum and from the pretectum to the thalamus. We suggest that the degeneration of visual axons might be produced by mechanical compression resulting from an increase in intraocular pressure.     

Purification to apparent homogeneity of the mating pheromone of mat-1 homozygous Euplotes raikovi

Mating type-specific mating pheromones autonomously released into the environment control cell-cell union in conjugation of the marine ciliate Euplotes raikovi. The mating pheromone, termed euplomone r-1, of cells of the homozygous mating type determined by the mat-1/mat-1 genotype was purified by means of a three-stage purification procedure which provides a yield of 79%. Starting from 10 liters of supernatant, 3.3 mg of euplomone r-1 were regularly recovered. Euplomone r-1 was identified with a protein showing a molecular weight of 12,000, an isoelectric point of 3.7, and unusually high contents of cysteine (15%) and tyrosine (5.8%). It appeared active at a concentration of 3.4 +/- 0.5 X 10(-12) M. Carbohydrate and a small colored compound, not yet identified, occurred in association with partially purified euplomone r-1 samples, whereas they were not detected in samples of the final product of purification. An acidic shift was apparently capable of destroying the carbohydrate/euplomone r-1 association, suggesting that it does not involve covalent bonds.