Decorin transfection induces proteomic and phenotypic modulation in breast cancer cells 8701-BC

Decorin is a prototype member of the small leucine-rich proteoglycan family widely distributed in the extracellular matrices of many connective tissues, where it has been shown to play multiple important roles in the matrix assembly process, as well as in some cellular activities. A major interest for decorin function concerns its role in tumorigenesis, as growth-inhibitor of different neoplastic cells, and potential antimetastatic agent. The aim of our research was to investigate wide-ranged effects of transgenic decorin on breast cancer cells. To this purpose we utilized the well-characterized 8701-BC cell line, isolated from a ductal infiltrating carcinoma of the breast, and two derived decorin-transfected clones, respectively, synthesizing full decorin proteoglycan or its protein core. The responses to the ectopic decorin production were examined by studying morphological changes, cell proliferation rates, and proteome modulation. The results revealed new important antioncogenic potentialities, likely exerted by decorin through a variety of distinct biochemical pathways. Major effects included the downregulation of several potential breast cancer biomarkers, the reduction of membrane ruffling, and the increase of cell-cell adhesiveness. These results disclose original aspects related to the reversion of malignant traits of a prototype of breast cancer cells induced by decorin. They also raise additional interest for the postulated clinical application of decori

A survey of grapevine fanleaf nepovirus isolates for the presence of satellite RNA

Grapevine fanleaf virus (GFLV) isolates from different geographical origins were surveyed for natural occurrence of satellite RNA. The results of molecular hybridization assays indicated that 5 isolates out of 34 tested, support the multiplication of a satellite RNA, both in Chenopodium quinoa and grapevine. The satellite molecules appear to have a high degree of sequence homology with, and the same size of, the satellite RNA supported by GFLV-F13 strain, isolated and characterized in France

Detection of grapevine closterovirus A in infected grapevine tissue by reverse transcription-polymerase chain reaction

Reverse transcription-polymerase chain reaction (RT-PCR) was successfully applied to detection of GVA RNA in nucleic acid extracts of infected grapevines. In particular, an artificially synthesized DNA primer set designed to amplify a GVA cDNA fragment of 430 base pairs, specifically detected GVA RNA sequences in extracts from infected grapevine tissues such as leaves from in vitro-grown explants, leaves from greenhouse-grown rooted cuttings, and bark scrapings of mature canes from field-grown vines. The detection limit of GVA RNA by RT-PCR was estimated to be 200 fold higher than that obtained by molecular hybridization or ELISA

Viroids of grapevines in Italy

Rebviroide in ItalienEs wird über das Vorkommen niedermolekularer RNAs bei 48 Vitis-vinifera-Stämmen und 15 amerikanischen Vitis-Arten und Kreuzungen aus Italien, Osteuropa, Mittelmeer- und Nahost-Ländern berichtet. In den Sämlingen zweier Sorten wurden keine derartigen RNAs gefunden. Aufgrund ihres elektrophoretischen Verhaltens wurden diese RNAs vorläufig identifiziert als Grapevine yellow speckle-Viroid (GYSVd), Grapeivine-Viroid 2 (GVd2) und Hop stunt-Viroid (HSVd). Das letztere Viroid löste bei künstlich infizierten Pflanzen von Tomate cv. Rutgers und Gurke cv. Suyo Befallssymptome aus. HSVd, GYSVd und GVd2 wurden in 97, 92 bzw. 11 % der untersuchten Proben wiedergefunden. In der Regel lagen Mischinfektionen vor, wobei die Kombination von HSVd mit GYSVd überwog. Diese beiden Viroide kamen regelmäßig in Reben mit den Symptomen von Yellow speckle oder Vein banding vor. Es wurde keine eindeutige Beziehung zwischen dem Vorkommen eines der Viroide und Rebkrankheiten mit unklarer Ätiologie, wie Vein necrosis oder Fleck, gefunden

Further studies on the use of molecular probes to grapevine closterovirus A

Two cloned cDNA probes to genomic RNA of grapevine closterovirus A (GVA) were utilized successfully for the detection of viral sequences in infected herbaceous hosts (Nicotiana benthamiana) and grapevines. One of the probes (pGA112) was complementary to the central part of the viral genome and gave light false positive signals with healthy grapevine extracts, whereas the other (pGA240), which is presumably colinear with the 3' terminus, was virus-specific and hybridized only with infected sample extracts. The two probes recognized smaller than genome RNAs in electrophoresed N. benthamiana extracts and hybridized differentially with the bands, thus suggesting that these represent subgenomic RNAs. Probe pGA240 may be used for GVA detection, but the preparation of samples for hybridization needs further improvement for routine testing.Weitere Untersuchungen über die Verwendung von Molekülsonden beim Grapevine-Closterovirus AZwei klonierte cDNA-Sonden für genomische RNA des Grapevine-Closterovirus A (GVA} wurden mit Erfolg zum Nachweis von Virussequenzen in infizierten krautigen Wirtspflanzen (Nicotiana benthamiana) und Reben benützt. Die eine Sonde (pGA112} war komplementär zum zentralen Teil des Virusgenoms und lieferte schwache falsch-positive Nachweisreaktionen mit Extrakten aus gesunden Reben; die andere Sonde (pGA240}, die vermutlich kolinear mit dem 3'-Terminus ist, war dagegen virusspezifisch und hybridisierte nur mit Extrakten aus infizierten Proben. In elektrophoretisch aufgetrennten N. benthamiana-Extrakten "erkannten" die beiden Sonden RNAs von weniger als Genomgröße; sie hybridisierten differenziert mit den Banden, so daß diese subgenomische RNAs darstellen könnten. Die Sonde pGA240 kann für den GVA-Nachweis verwendet werden; für Routinetests muß die Vorbereitung der Proben für die Hybridisierung noch verbessert werden

Stable insertion and expression of the movement protein gene of Grapevine Virus A (GVA) in grape (Vitis rupestris S.)

Transformation with the movement protein gene of Grapevine virus A, both in sense and antisense orientation, was done in Vitis rupestris S. somatic embryos through LBA 4404 Agrobacterium tumefaciens co-cultures, and plantlets were regenerated. Molecular assays of regenerated plantlets, after 4 years of micropropagation cycles, verified stable insertion and expression of the foreign genes in both orientations. Plants expressing the sense form of the viral gene, exhibited morphological and physiological anomalies, such as slow growth, suppression of buds and flowering and tendril development.

Photoluminescence properties of C60 films deposited on silicon substrate

Photoluminescence (PL) spectra of C-60 films deposited on Si substrates have been measured from 10 to 300 K and as a function of laser excitation intensity. Recombination of self-trapped excitons and their phonon replicas, as well as X-trap-related emissions, are the main features of the PL spectra. The influence of the deposition parameters, namely deposition rate and substrate temperature, on the luminescence efficiency of the C-60 films have been investigated. Low substrate temperature produces a lowering of the PL efficiency, whereas an increase of the deposition rate causes an increase of the X-trap emission

Serological detection of Grapevine rupestris stem pitting-associated virus (GRSPa V) by a polyclonal antiserum to recombinant virus coat protein

The coat protein gene of Grapevine rupestris stem pitting-associated virus (GRSPaV) was amplified with primers based on the completely sequenced Californian GRSPaV isolate, The protein expressed in Escherichia coli was used to raise an antiserum in rabbit. This antiserum was successfully used to detect virus coat protein in infected grapevine extracts either spotted on polyvinyl difluoride membranes (dot immunobinding) or blotted on membranes after gel separation (Western blot). The antiserum titre was 1:5,000 in Western blot. GRSPaV was detected in leaf petioles and cortical scrapings from dormant canes during the whole vegetative season. Several accessions of Vitis rupestris, currently used as presumptive virus-free indicators of Rupestris stem pitting, were found to be infected by this virus. While the application of the antiserum in ELISA was ineffective, the availability of similarly simple and effective serological tools, such as dot immunobinding, may allow a wide survey for GRSPaV

NASA's Solar System Exploration Research Virtual Institute: Merging Science and Exploration

NASA's Solar System Exploration Research Virtual Institute (SSERVI) represents a close collaboration between science, technology and exploration, and was created to enable a deeper understanding of the Moon and other airless bodies. SSERVI is supported jointly by NASA's Science Mission Directorate and Human Exploration and Operations Mission Directorate. The institute currently focuses on the scientific aspects of exploration as they pertain to the Moon, Near Earth Asteroids (NEAs) and the moons of Mars, but the institute goals may expand, depending on NASA's needs, in the future. The 9 initial teams, selected in late 2013 and funded from 2014-2019, have expertise across the broad spectrum of lunar, NEA, and Martian moon sciences. Their research includes various aspects of the surface, interior, exosphere, near-space environments, and dynamics of these bodies. NASA anticipates a small number of additional teams to be selected within the next two years, with a Cooperative Agreement Notice (CAN) likely to be released in 2016. Calls for proposals are issued every 2-3 years to allow overlap between generations of institute teams, but the intent for each team is to provide a stable base of funding for a five year period. SSERVI's mission includes acting as a bridge between several groups, joining together researchers from: 1) scientific and exploration communities, 2) multiple disciplines across a wide range of planetary sciences, and 3) domestic and international communities and partnerships. The SSERVI central office is located at NASA Ames Research Center in Mountain View, CA. The administrative staff at the central office forms the organizational hub for the domestic and international teams and enables the virtual collaborative environment. Interactions with geographically dispersed teams across the U.S., and global partners, occur easily and frequently in a collaborative virtual environment. This poster will provide an overview of the 9 current US teams and international partners, as well as information about outreach efforts and future opportunities to participate in SSERVI

Worldwide diffusion of Fig latent virus 1 in fig accessions and its detection by serological and molecular tools

A virus with filamentous particles ca. 700 nm long, denoted Fig latent virus 1 (FLV-1) is widespread in Apulian (southern Italy) fig orchards, in trees showing or not mosaic symptoms and in symptomless seedlings. The virus was transmitted by sap inoculation to a very restricted range of herbaceous hosts without inducing apparent symptoms and was transmitted through fig seeds to a very high percentage (80 to 100 %). It was successfully purified from root tissues of infected figs. A virus-specific antiserum raised in rabbits, proved useful for its detection in fig leaf dips by immunosorbent electron microscopy (ISEM), Western Blot, dot immuno-binding (DIBA), ELISA. The viral genome structure resembles that of members of the genus Trichovirus in the family Flexiviridae. Keywords: fig latent virus, Trichovirus, serology, ISEM, Western blot, DIBA, ELIS