Fibulin-2 stabilizes tumor extracellular matrix and drives malignant progression of lung adenocarcinoma

The ECM of epithelial carcinomas undergoes structural remodeling during periods of uncontrolled growth, creating regional heterogeneity and torsional stress. How tumors maintain ECM integrity in the face of dynamic biophysical forces is still largely unclear. This study addresses these deficiencies using mouse models of human lung adenocarcinoma. Spontaneous lung tumors were marked by disorganized basement membranes, dense collagen networks, and increased tissue stiffness. Metastasis-prone lung adenocarcinoma cells secreted fibulin-2 (Fbln2), a matrix glycoprotein involved in ECM supra-molecular assembly. Fibulin-2 depletion in tumor cells decreased the intra-tumoral abundance of matrix metalloproteinases and reduced collagen cross-linking and tumor compressive properties resulting in inhibited tumor growth and metastasis. Fbln2 deposition within intra-tumoral fibrotic bands was a predictor of poor clinical outcome in patients. Collectively, these findings support a feed-forward model in which tumor cells secrete matrix-stabilizing factors required for the assembly of ECM that preferentially favors malignant progression. To our knowledge, this is the first evidence that tumor cells directly regulate the integrity of their surrounding matrix through the secretion of matrix-stabilizing factors such as fibulin-2. These findings open a new avenue of research into matrix assembly molecules as potential therapeutic targets in cancer patients.

Interactions and structural analysis of the zinc -binding motif in decorin, a small leucine rich proteoglycan in extracellular matrix

Decorin, a dermatan/chondroitin sulfate proteoglycan, is ubiquitously distributed in the extracellular matrix (ECM) of mammals. Decorin belongs to the small leucine rich proteoglycan (SLRP) family, a proteoglycan family characterized by a core protein dominated by Leucine Rich Repeat motifs. The decorin core protein appears to mediate the binding of decorin to ECM molecules, such as collagens and fibronectin. It is believed that the interactions of decorin with these ECM molecules contribute to the regulation of ECM assembly, cell adhesions, and cell proliferation. These basic biological processes play critical roles during embryonic development and wound healing and are altered in pathological conditions such as fibrosis and tumorgenesis. ^ In this dissertation, we discover that decorin core protein can bind to Zn2+ ions with high affinity. Zinc is an essential trace element in mammals. Zn2+ ions play a catalytic role in the activation of many enzymes and a structural role in the stabilization of protein conformation. By examining purified recombinant decorin and its core protein fragments for Zn2+ binding activity using Zn2+-chelating column chromatography and Zn2+-equilibrium dialysis approaches, we have located the Zn2+ binding domain to the N-terminal sequence of the decorin core protein. The decorin N-terminal domain appears to contain two Zn2+ binding sites with similar high binding affinity. The sequence of the decorin N-terminal domain does not resemble any other reported zinc-binding motifs and, therefore, represents a novel Zn 2+ binding motif. By investigating the influence of Zn2+ ions on decorin binding interactions, we found a novel Zn2+ dependent interaction with fibrinogen, the major plasma protein in blood clots. Furthermore, a recombinant peptide (MD4) consisting of a 41 amino acid sequence of mouse decorin N-terminal domain can prolong thrombin induced fibrinogen/fibrin clot formation. This suggests that in the presence of Zn2+ the decorin N-terminal domain has an anticoagulation activity. The changed Zn2+-binding activities of the truncated MD4 peptides and site-directed mutagenesis generated mutant peptides revealed that the functional MD4 peptide might contain both a structural zinc-binding site in the cysteine cluster region and a catalytic zinc site that could be created by the flanking sequences of the cysteine cluster region. A model of a loop-like structure for MD4 peptide is proposed. ^

Gene clustering of the small leucine -rich repeat gene family

The small leucine-rich repeat proteoglycans (or SLRPs) are a group of extracellular proteins (ECM) that belong to the leucine-rich repeat (LRR) superfamily of proteins. The LRR is a protein folding motif composed of 20–30 amino acids with leucines in conserved positions. LRR-containing proteins are present in a broad spectrum of organisms and possess diverse cellular functions and localization. In mammals, the SLRPs are abundant in connective tissues, such as bones, cartilage, tendons, skin, and blood vessels. We have discovered a new member of the class I small leucine rich repeat proteoglycan (SLRP) family which is distinct from the other class I SLRPs since it possesses a unique stretch of aspartate residues at its N-terminus. For this reason, we called the molecule asporin. The deduced amino acid sequence is about 50% identical (and 70% similar) to decorin and biglycan. However, asporin does not contain a serine/glycine dipeptide sequence required for the assembly of O-linked glycosaminoglycans and is probably not a proteoglycan. The tissue expression of asporin partially overlaps with the expression of decorin and biglycan. During mouse embryonic development, asporin mRNA expression was detected primarily in the skeleton and other specialized connective tissues; very little asporin message was detected in the major parenchymal organs. The mouse asporin gene structure is similar to that of biglycan and decorin with 8 exons. The asporin gene is localized to human chromosome 9q22-9g21.3 where asporin is part of a SLRP gene cluster that includes ECM2, osteoadherin, and osteoglycin. This gene cluster of four LRR-encoding genes is embedded in a 238 kilobase intron of another novel gene named Tes9orf that is expressed primarily in the testes of the adult mouse. The SLRP genes are not present in Drosophila or C. elegans , but reside in three separate gene clusters in the puffer fish, mice and humans. Targeted disruption of individual mouse SLRP genes display minor connective tissue defects such as skin fragility, tendon laxity, minor growth plate defects, and mild osteoporosis. However, double and triple knockouts of SLRP genes exacerbate these phenotypes. Both the double epiphycan/biglycan and the triple PRELP/fibromodulin/biglycan knockout mice exhibit premature osteoarthritis. ^

YKL -40, a vascular -associated serum marker for glioblastoma, correlates with chronic activation of AKT

YKL-40 is a secreted glycoprotein that has been reported to be expressed in pathologic conditions of extracellular matrix degradation and angiogenesis, such as rheumatoid arthritis, severe osteoarthritis, primary colorectal cancer, metastatic breast cancer, and recurrent ovarian cancer (Dehn, Hogdall et al. 2003). ^ We have identified YKL-40 as a serum marker for glioblastoma multiforme (GBM) using microarray analysis from samples of GBM. We compared the gene expression profile of 19 gliomas to pooled normal brain tissue using the Incyte 10,000 gene expression array. The most differentially expressed gene in this analysis was YKL-40; it was detected in GBM samples with a range of 3 to 62-fold elevation over normal brain. Western blot analysis of glioma samples for YKL-40 protein levels revealed substantial elevation in approximately 65% of GBMs, and undetectable levels in lower-grade gliomas and normal brain tissue. ELISA analysis on serum samples of glioma patients showed that YKL-40 levels were substantially elevated in many of the GBM patients. Statistical analysis indicated that in patients with glioma, serum YKL-40 levels correlate with tumor grade and potentially tumor burden in GBM. ^ Furthermore, we found that YKL-40 expression by in-situ hybridization on a brain tumor tissue array was limited to GBM's and gliosarcomas (GSA), and that YKL-40 expression was specific to the GBM component of GSA. Additional in-situ hybridization analysis, found it to be regionally associated with tumor vasculature as well as activated AKT expression in both human and mouse GBM's. Correlation of elevated YKL-40 with phospho-AKT was confirmed by Western blot analysis on a series of glioblastoma tumors, and inhibition of PI3 Kinase signaling by addition of LY294002 also decreased secretion of YKL-40 over a 7-day period in U87 glioblastoma cell tine. Lastly, YKL-40 expression was induced in response to serum starvation and altered by interaction with specific extracellular matrix (ECM) modules. In summary, we have identified the first accurate serum marker for high-grade gliomas. Furthermore, our findings indicate that YKL-40 is a highly expressed vascular-related glycoprotein in human GBM tissue and that it is affected by the AKT signaling pathway and interaction with components of brain ECM proteins. ^

Specific conductivity and hydrogen ions measured on two ice core (NEEM and NGRIP)

A stratigraphy-based chronology for the North Greenland Eemian Ice Drilling (NEEM) ice core has been derived by transferring the annual layer counted Greenland Ice Core Chronology 2005 (GICC05) and its model extension (GICC05modelext) from the NGRIP core to the NEEM core using 787 match points of mainly volcanic origin identified in the electrical conductivity measurement (ECM) and dielectrical profiling (DEP) records. Tephra horizons found in both the NEEM and NGRIP ice cores are used to test the matching based on ECM and DEP and provide five additional horizons used for the timescale transfer. A thinning function reflecting the accumulated strain along the core has been determined using a Dansgaard-Johnsen flow model and an isotope-dependent accumulation rate parameterization. Flow parameters are determined from Monte Carlo analysis constrained by the observed depth-age horizons. In order to construct a chronology for the gas phase, the ice age-gas age difference (Delta age) has been reconstructed using a coupled firn densification-heat diffusion model. Temperature and accumulation inputs to the Delta age model, initially derived from the water isotope proxies, have been adjusted to optimize the fit to timing constraints from d15N of nitrogen and high-resolution methane data during the abrupt onset of Greenland interstadials. The ice and gas chronologies and the corresponding thinning function represent the first chronology for the NEEM core, named GICC05modelext-NEEM-1. Based on both the flow and firn modelling results, the accumulation history for the NEEM site has been reconstructed. Together, the timescale and accumulation reconstruction provide the necessary basis for further analysis of the records from NEEM.

Density and stable oxygen isotope profiles of four snow pits from Berkner Island, Filchner-Ronne Ice Shelf, Antarctica

The ice cap on Berkner Island is grounded on bedrock within the Filchner-Ronne Ice Shelf and is, therefore, expected to be a well-suited place to retrieve long-term ice-core records reflecting the environmental situation of the Weddell Sea region. Shallow firn cores were drilled to 11 m at the two main summits of Berkner Island and analysed in high depth resolution for electrical d.c. conductivity (ECM), stable isotopes, chloride, sulphate, nitrate and methane-sulphonate (MSA). From the annual layering of dD and non-sea-salt (nss) sulphate, a mean annual snow accumulation of 26.6 cm water at the north dome and 17.4 cm water at the south dome are obtained. As a result of ineffective wind scouring indicated by a relatively low near-surface snow density, regular annual cycles are found for all species at least in the upper 4-5 m. Post depositional changes are responsible for a substantial decrease of the seasonal dD and nitrate amplitude as well as for considerable migration of the MSA signal operating below a depth of 3-4 m. The mean chemical and isotopic firn properties at the south dome correspond to the situation on the Filchner-Ronne Ice shelf at a comparable distance to the coast, whereas the north dome is found to be more influenced by maritime air masses. Persistent high sea-salt levels in winter snow at Berkner Island heavily obscure the determination of nss sulphate probably due to sulphate fractionation in the Antartic sea-salt aerosols. Estimated time-scales predict ages at 400 m depth to be ca. 2000 years for the north and ca. 3000 years for the south dome. Pleistocene ice is expected in the bottom 200 and 300 m, respectively.

Conductivity and oxygen isotope measured on ice core FRI09C90_13 (B13) from the Filchner-Ronne ice shelf, Antarctica

The Filchner-Ronne ice shelf, which drains most of the marine-based portions of the West Antarctic ice sheet, is the largest ice shelf on Earth by volume. The origin and properties of the ice that constitutes this shelf are poorly understood, because a strong reflecting interface within the ice and the diffuse nature of the ice?ocean interface make seismic and radio echo sounding data difficult to interpret. Ice in the upper part of the shelf is of meteoric origin, but it has been proposed that a basal layer of saline ice accumulates from below. Here we present the results of an analysis of the physical and chemical characteristics of an ice core drilled almost to the bottom of the Ronne ice shelf. We observe a change in ice properties at about 150 m depth, which we ascribe to a change from meteoric ice to basal marine ice. The basal ice is very different from sea ice formed at the ocean surface and we propose a formation mechanism in which ice platelets in the water column accrete to the bottom of the ice shelf.

Continuous density log of icecore BER11C95_25

A 181 m long ice core was drilled at 79° 36' 51'' S, 45° 43' 28'' W, near the summit of Berkner Island, Antarctica 886 m a.s.l.). Berkner Island is located within the Filchner and Ronne Ice Shelves, and the ice near the summit shows little lateral flow. The density of the ice core was measured every 3 mm along ist length, using attenuation of a gamma-ray beam, which gave an absolute accuracy of 2%. As expected, there is a general density increase with depth, the maximum densities of > 900 kg/m**3 being reached just above 100 m depth. Comparison with electrical conductivity method (ECM) shows density variations with the same wavelength as annual signals, which can be seen in the ECM log (higher acidity during summer). In the shallowest part of the core, the density of winter layers is higher than that of summer layers, a relationship which is reversed at greater depth. We assume that the densification rates for the two types of firn are different. Similar density phenomena were observed on ice cores from Greenland, showing that such phenomena are not a local effect.

Physical and chemical analysis of firn cores from Ronne Ice Shelf, Antarctica

This data on the distribution of the accumulation rate and 18O content of near-surface layers in the eastern part of the Ronne Ice Shelf, Antarctica, were derived from an analysis of 16 firn cores. The firn cores were drilled along the traverse route of the Filchner-V-Campaign in 1995. The traverse followed an ice flowline of the Foundation Ice Stream and reached the margin of the inland ice, an area which has not yet been investigated. On the ice shelf the accumulation rates decrease with distance from the coast. Ascending to the inland ice the accumulation rates again reach almost coastal values. This regional distribution is in agreement with the temperature gradient along the traverse. The 18O content of the near-surface layers is closely related to the 10 m firn temperature. They strongly decrease from the grounding line towards the inland ice. At the southernmost site at 1100 m a.s.l., the mean d18O value of the firn decreases to -40?. Ice with that isotopic signature was found in cores from the central part of the Ronne Ice Shelf just above the marine ice layer, indicating that it originates from this area. All ice deposited as snow further south was melted beneath the ice shelf after passing the grounding-line area. The time series of accumulation rate and 18O content reveal no climatic trend during the last 30-50 years.

1000 year ice-core records from Berkner Island, Antarctica

Two medium-depth ice cores were retrieved from Berkner Island by a joint project between the Alfred-Wegener-Institut and the British Antarctic Survey in the 1994/95 field season. A 151m deep core from the northern dome (Reinwarthhöhe) of Berkner Island spans 700 years, while a 181m deep core from the southern dome (Thyssenhöhe) spans approximately 1200 years. Both cores display clear seasonal cycles in electrical conductivity measurements, allowing dating by annual-layer counting and the calculation of accumulation profiles. Stable-isotope measurements (both d18O and dD), together with the accumulation data, allow us to estimate changes in climate for most of the past millennium: the data show multi-decadal variability around a generally stable long-term mean. In addition, a full suite of major chemistry measurements is available to define the history of aerosol deposition at these sites: again, there is little evidence that the chemistry of the sites has changed over the past six centuries. Finally, we suggest that the southern dome, with an ice thickness of 950 m, is an ideal site from which to gain a climate history of the late stages of the last glacial and the deglaciation for comparison with the records from the deep Antarctic ice cores, and with other intermediate-depth cores such asTaylor Dome and Siple Dome.

Diseño electrónico de la tarjeta de interfaz y control para un equipo de seguimiento en elevación en un subsistema de contramedidas electrónicas

Como apoyo al Subsistema ECM tradicional de los sistemas de Guerra Electrónica, surge el Equipo de Seguimiento en Elevación cuyo fin es la búsqueda de la amenaza en elevación. Ante la necesidad de un mayor número de líneas entre los bloques internos de este Equipo de Seguimiento, se hace necesario el diseño de una Tarjeta de Interfaz y Control. La tarjeta se ocupará de realizar el control de datos y flujo de señales de módulos RF que componen el Equipo, haciendo de interfaz entre las Tarjetas Procesadoras y Digitalizadoras, así como entre la Tarjeta de Proceso y el Bloque de Recepción. Una vez que se haya realizado el análisis de la necesidad de controlar estas señales, estudiado las especificaciones y los requisitos funcionales del Sistema, se procederá a implementar el diseño hardware y desarrollo firmware de la Tarjeta de Interfaz y Control de un Equipo de Seguimiento en Elevación para un Subsistema ECM, que constituye el objetivo de este PFC. ABSTRACT. In order to find a threat on elevation, and as a support on the basic ECM subsystems of E-war systems, the Elevation Tracking Equipment appears. To the need of a bigger number of lines between inside blocks of this Tracking Equipment, the design of an Interface and Control Board is needed. This board will deal with the data control and with the flow of RF modules signals that set up the Equipment, and will also work as an interface between the Processing and Digitalizing Board, as well as between the Process Board and the Reception Block. Once the signal control necessity analysis has been made, and once the specifications and functional System requirements have been studied, the hardware design and the firmware development will be ran as a target of PFC.