Proteomic patterns of cultured breast cancer cells and epithelial mammary cells.

Breast cancer is one of the leading causes of death from cancer among women in western countries. The different types of breast cancer are grouped into invasive and noninvasive forms. Among the invasive types, ductal infiltrating carcinoma (DIC) is the most common and aggressive form. Using an in vitro model consisting of a DIC-derived cell line (8701-BC) and a nontumoral mammary epithelial cell line (HB2), we used the proteomics approach to search for homology and differences in protein expression patterns between tumoral and nontumoral phenotypes. Within an analysis window comprising 1,750 discernible spots we have currently catalogued 140 protein spots of potential interest. Fifty-eight of them were identified by gel matching with reference maps, immunodetection, or N-terminal microsequencing and classified into four functional groups. Twelve proteins were found differentially expressed in two cell lines: four were uniquely present in the neoplastic cell proteome and eight in epithelial cells. In addition, 53 proteins displayed different relative expression levels between the two cell lines, that is, 44 were more elevated in cancer cells and 9 in HB2 cells. Among proteins with greater relative abundance in cancer cells we identified glycolytic enzymes (or their isoforms), which may indicate that the known metabolic dysregulation in cancer can reflect oncogenic-related defects of glycolytic gene expression

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

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

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

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

Caveolin-1, breast cancer and ionizing radiation

Breast cancer (BC) recovery has increased in recent years thanks to efforts of Omics-based research in this field. However, despite the important results obtained, BC remains a complex multifactorial pathology that is difficult to treat appropriately. Caveolin-1 (CAV1), the basic constituent protein of specialized plasma membrane invaginations called caveolae, is emerging as a potential therapeutic biomarker in BC. This factor may modulate BC response to chemotherapy and radiation therapy. In addition, recent reports describe the key role of CAV1 during cell response to oxidative stress. The aim of the present review was to describe the biological roles of CAV1 in BC considering its contrasting dual functions as an oncogene and as a tumor suppressor. In addition, we report on how CAV1 may contribute to tumor cell response to ionizing radiation treatment. Finally, new roles of CAV1 in BC both on epithelium and stroma may be useful as prognostic indicators for patient treatment and help clinicians in the selection of the best personalized therapy

Integrated multi-omics investigations of metalloproteinases in colon cancer: Focus on MMP2 and MMP9

Colorectal cancer (CRC) develops by genetic and epigenetic alterations. However, the molecular mechanisms underlying metastatic dissemination remain unclear and could benefit from multi-omics investigations of specific protein families. Matrix metalloproteinases (MMPs) are proteolytic enzymes involved in ECM remodeling and the processing of bioactive molecules. Increased MMP expression promotes the hallmarks of tumor progression, including angiogenesis, invasion, and metastasis, and is correlated with a shortened survival. Nevertheless, the collective role and the possible coordination of MMP members in CRC are poorly investigated. Here, we performed a multi-omics analysis of MMP expression in CRC using data mining and experimental investigations. Several databases were used to deeply mine different expressions between tumor and normal tissues, the genetic and epigenetic alterations, the prognostic value as well as the interrelationships with tumor immune-infiltrating cells (TIICs). A special focus was placed on to MMP2 and MMP9: their expression was correlated with immune markers and the interaction network of co-expressed genes disclosed their implication in epithelial to mesenchymal transition (EMT) and immune response. Finally, the activity levels of MMP2 and MMP9 in a cohort of colon cancer samples, including tissues and the corresponding sera, was also investigated by zymography. Our findings suggested that MMPs could have a high potency, as they are targeted in colon cancer, and might serve as novel biomarkers, especially for their involvement in the immune response. However, further studies are needed to explore the detailed biological functions and molecular mechanisms of MMPs in CRC, also in consideration of their expression and different regulation in several tissues

Retrospective Proteomic Screening of 100 Breast Cancer Tissues

The present investigation has been conducted on one hundred tissue fragments of breast cancer, collected and immediately cryopreserved following the surgical resection. The specimens were selected from patients with invasive ductal carcinoma of the breast, the most frequent and potentially aggressive type of mammary cancer, with the objective to increase the knowledge of breast cancer molecular markers potentially useful for clinical applications. The proteomic screening; by 2D-IPG and mass spectrometry; allowed us to identify two main classes of protein clusters: proteins expressed ubiquitously at high levels in all patients; and proteins expressed sporadically among the same patients. Within the group of ubiquitous proteins, glycolytic enzymes and proteins with anti-apoptotic activity were predominant. Among the sporadic ones, proteins involved in cell motility, molecular chaperones and proteins involved in the detoxification appeared prevalent. The data of the present study indicates that the primary tumor growth is reasonably supported by concurrent events: the inhibition of apoptosis and stimulation of cellular proliferation, and the increased expression of glycolytic enzymes with multiple functions. The second phase of the evolution of the tumor can be prematurely scheduled by the occasional presence of proteins involved in cell motility and in the defenses of the oxidative stress. We suggest that this approach on large-scale 2D-IPG proteomics of breast cancer is currently a valid tool that offers the opportunity to evaluate on the same assay the presence and recurrence of individual proteins, their isoforms and short forms, to be proposed as prognostic indicators and susceptibility to metastasis in patients operated on for invasive ductal carcinoma of the breast

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