Maintenance of EGFR and EGFRvIII expressions in an in vivo and in vitro model of human glioblastoma multiforme

Glioblastoma multiforme (GBM) is the most common, and most aggressive primary brain tumor among adults. A vast majority of the tumors express high levels of the epidermal growth factor receptor (EGFR) as a consequence of gene amplification. Furthermore, gene amplification is often associated with mutation of EGFR, and the constitutive activated deletion variant EGFRvIII is the most common EGFR mutation found in GBM. Activated EGFR signaling, through overexpression and/or mutation, is involved in increased tumorigenic potential. As such, EGFR is an attractive target for GBM therapy. However, clinical studies with EGFR inhibitors have shown inconsistent results, and as such, further knowledge regarding the role of EGFR and EGFRvIII in GBM is needed. For this, an appropriate in vivo/in vitro tumor model is required. Here, we report the establishment of an experimental GBM model in which the expressions of EGFR and EGFRvIII are maintained both in xenograft tumors growing subcutaneously on mice and in cell cultures established in stem cell conditions. With this model it will be possible to further study the role of EGFR and EGFRvIII, and response to targeted therapy, in GBM.     

Plasmid segregation: spatial awareness at the molecular level

In bacteria, low-copy number plasmids ensure their stable inheritance by partition loci (par), which actively distribute plasmid replicates to each side of the cell division plane. Using time-lapse fluorescence microscopic tracking of segregating plasmid molecules, a new study provides novel insight into the workings of the par system from Escherichia coli plasmid R1. Despite its relative simplicity, the plasmid partition spindle shares characteristics with the mitotic machinery of eukaryotic cells.     

Spatial and temporal variability of the benthic assemblages associated to the holdfasts of the kelp <Emphasis Type="Italic">Macrocystis pyrifera</Emphasis> in the Straits of Magellan,Chile

Macrocystis pyrifera (L.) C. Agardh is a characteristic macroalga in the Magellan region covering almost 30% of the shallow coastal waters. The focus of this study was to analyse the spatial and seasonal patterns in macrofauna communities associated to the holdfasts of Macrocystis pyrifera at two study sites in the Straits of Magellan, South Chile. In total, 114 species from 10 major taxa were isolated from the holdfasts. MDS clearly separated the holdfast fauna collected in different seasons, with autumn and winter collections being richer in terms of species richness and abundance as compared to the spring and summer situation. MDS also clearly separated the holdfast associated faunas of the two study sites, Bahía Laredo and Fuerte Bulnes. The community structure and species composition of the associated macro-invertebrates and vertebrates appeared rather heterogeneous, probably due to the extremely heterogeneous environmental conditions along the entire coastline of the Subantarctic Magellan region.     

Upregulation of gamma-catenin compensates for the loss of beta-catenin in adult cardiomyocytes

Recent progresses in signal transduction have revealed that beta-catenin signaling controls embryonic development, tumorigenesis, cell shape, and polarity. The role of this pathway in myocyte shape regulation during cardiac hypertrophy and failure is, however, not clearly defined. Since homozygous knockout of beta-catenin is embryonically lethal, we have deleted beta-catenin genes specifically in the heart of adult mice by crossing loxP-flanked beta-catenin mice with transgenic mice expressing tamoxifen-activated MerCreMer protein (MCM) driven by the alpha-myosin heavy chain promoter. Administration of tamoxifen to homozygous loxP-flanked beta-catenin mice positive for MCM induces the deletion of beta-catenin only in cardiomyocytes. Immunolabeling with beta-catenin antibody demonstrates that 90% of cardiomyocytes completely lose their beta-catenin expression but maintain normal rod-shaped morphology. The intercalated disk of cardiomyocytes lacking beta-catenin is morphologically unremarkable with normal distribution of vinculin, N-cadherin, desmoplakin, ZO-1, connexin43, and alpha-, gamma-, and p120 catenins. The expression level of these proteins, except that of gamma-catenin, is also similar in tamoxifen-treated and control mice with both homozygous loxP-flanked beta-catenin genes and the MCM transgene. Western blot analyses reveal that gamma-catenin increases in the heart of beta-catenin knockout mice compared with controls. Confocal microscopy also demonstrates that gamma-catenin has significantly increased in the intercalated disk of cardiomyocytes lacking beta-catenin. Echocardiographic data indicate that the knockout mice maintain normal ventricular geometry and cardiac function. The results suggest that upregulation of gamma-catenin can compensate for the loss of beta-catenin in cardiomyocytes to maintain normal cardiac structure and function.     

Decreased Expression of Cilia Genes in Pancreatic Islets as a Risk Factor for Type 2 Diabetes in Mice and Humans

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Bryozoans and microbial communities of cool-temperate to subtropical latitudes—paleoecological implications

The environmental distribution of encrusting bryozoans settling on disarticulated and living bivalve shells has been recorded from five stations in Japan and New Zealand. Some insight into the observed distribution patterns emerges from information on the interaction of bryozoans with microbial mats. Advancing existing classifications, we have subdivided the encrusting bryozoan morphotypes into seven different growth types that largely reflect the biological potentials of bryozoans in competition for space on substrate surfaces. The frequency distribution of these types (s-/c-/m-/z-laminae, runners, spots, bryostromatolites) reveals the influence of microbial mats as a control factor of bryozoan substrate coverage. Microbial mats in turn are correlated with latitudinal gradients in Japan and New Zealand from cool-temperate to subtropical and tropical waters. Unlike erect bryozoans, laminar ones are probably underrated as facies fossils. Accordingly, laminar bryozoan growth types are reconsidered as a tool for paleoecological interpretation of marine hard substrate communities.     

Dimerization of endogenous MT1-MMP is a regulatory step in the activation of the 72-kDa gelatinase MMP-2 on fibroblasts and fibrosarcoma cells

The secreted gelatinase matrix metalloprotease-2 (MMP-2) and the membrane-anchored matrix metalloprotease MT1-MMP (MMP-14), are central players in pericellular proteolysis in extracellular matrix degradation. In addition to possessing a direct collagenolytic and gelatinolytic activity, these enzymes take part in a cascade pathway in which MT1-MMP activates the MMP-2 proenzyme. This reaction occurs in an interplay with the matrix metalloprotease inhibitor, TIMP-2, and the proposed mechanism involves two molecules of MT1-MMP in complex with one TIMP-2 molecule. We provide positive evidence that proMMP-2 activation is governed by dimerization of MT1-MMP on the surface of fibroblasts and fibrosarcoma cells. Even in the absence of transfection and overexpression, dimerization of MT1-MMP markedly stimulated the formation of active MMP-2 products. The effect demonstrated here was brought about by a monoclonal antibody that binds specifically to MT1-MMP as shown by immunofluorescence experiments. The antibody has no effect on the catalytic activity. The effect on proMMP-2 activation involves MT1-MMP dimerization because it requires the divalent monoclonal antibody, with no effect obtained with monovalent Fab fragments. Since only a negligible level of proMMP-2 activation was obtained with MT1-MMP-expressing cells in the absence of dimerization, our results identify the dimerization event as a critical level of proteolytic cascade regulation.     

Carbonate ooids of the Mesoarchaean Pongola Supergroup,South Africa

Ooids from the Mesoarchaean Chobeni Formation, Pongola Supergroup, KwaZulu‐Natal, South Africa are older than any ooids reported to date. They are made of dolomite and ankerite and show concentric, radial‐concentric, micritic, and radial fabrics. Radial ooids are interpreted to have originated from high‐Mg‐calcite and probably formed by microbial activity in a low‐energy regime, while concentric ooids had an aragonite precursor and formed biotically under agitated/high‐energy environmental conditions. Micritic ooids formed via the recrystallization of concentric ooids. Ooids and other allochems, such as intraclasts and peloids, contain carbonaceous matter. The close association of carbonaceous matter within ooid cortices with metabolically important elements, such as N, S and P, as identified by nano‐scale secondary ion mass spectrometry analysis, allows us to propose a biologically induced origin for some ooids. By analogy with modern examples, a variety of microbial communities probably played a role in carbonate precipitation and ooid formation. Shale‐normalized rare earth element (REE) distribution patterns of ooids and other allochems show positive La_SN, Gd_SN and Y_SN anomalies, superchondritic Y/Ho ratios and depleted light rare earth elements (LREEs) relative to the heavy rare earth elements (HREEs), which resemble those of seawater. These anomalies are less pronounced than expected for an open marine setting, which is interpreted as evidence for deposition in restricted shallow marine environments. Non‐seawater REE patterns in recrystallized matrix and pore‐ and vein‐filling carbonate likely reflect redistribution of rare earth elements during post‐depositional alteration and/or reflect differences in the elemental and REE compositions of diagenetic fluids.     

Meiofauna response to iceberg disturbance on the Antarctic continental shelf at Kapp Norvegia (Weddell Sea)

The impact of iceberg scouring on meiofauna communities, especially nematodes, was studied on the Kapp Norvegia shelf in the Weddell Sea, Antarctica. Three stations with different stages of recolonisation following scour were selected on the basis of seafloor video images, sediment characteristics and faunal occurrences. These stations comprised a fresh scour, an older scour, and an undisturbed control site where a sponge spicule mat covered the sediment with dense epifauna. Meiofaunal abundance and taxonomic diversity of meiofauna groups were significantly reduced in the fresh scour. The highest abundance and diversity were found in the older scour as compared with the undisturbed site. The abundance and diversity of nematodes also decreased due to scouring. The abundance in the older scour recovered to the level of the undisturbed site whereas the diversity remained low. Scouring also changed the nematode community structure, with the suborders Desmoscolecina and Leptolaimina as the most sensitive groups. In addition, scouring resulted in the decrease of selective deposit feeders and the Maturity Index. The low diversity and the change in nematode generic composition in the older scour compared with the undisturbed site, despite the complete recovery in terms of abundance, suggest that the deep continental shelf nematode community in this area is sensitive to iceberg disturbance.     

A seafloor crater in the German Bight and its effects on the benthos

 In 1963 a deep crater was formed about 65 m below sea level in the western part of the German Bight, due to a gas eruption caused by drilling carried out from the platform ’Mr. Louie’. The study area is situated in a sandy to muddy bottom area inhabited by an Amphiura filiformis association (sensu Salzwedel et al. 1985). The crater, sometimes called ’Figge-Maar’, functions as a sediment trap, concentrating particles and organisms from the water column, thus leading to extreme sedimentation rates of about 50 cm, on average, per year. Crater stations, compared with stations situated in the vicinity, show enrichments of juveniles. Echinoderms, especially the subsurface-dwelling heart urchin Echinocardium cordatum and ophiuroids are responsive to enrichment. Other species that are typical of the Amphiura filiformis association are shown to be unable to cope with the special conditions in the crater. Received: 12 June 1998 / Accepted:16 September 1998