Casein production during differentiation of mammary epithelial cells in collagen gel culture

Mouse mammary epithelial cells cultivated on floating collagen gels secrete, as judged by immunoblotting, the full array of caseins found in mouse milk. The secreted caseins are all phosphorylated and have estimated minimum molecular weights (MWs) of 45, 40, 27, and 23 kD in SDS-PAGE. Intracellular caseins of epithelia from collagen gel cultivation or from lactating mammary glands are a combination of mature caseins identical with the secreted molecules and novel caseins whose apparent size in SDS-PAGE is different from the secreted molecules. The novel caseins were shown to be non-phosphorylated species apparently insufficiently mature for secretion. Our data indicate that, with regard to casein expression, cultivation of mouse mammary epithelia on collagen gels essentially duplicates their behavior in the lactating mouse mammary glands.     

Biofilm formation behaviour of marine filamentous cyanobacterial strains in controlled hydrodynamic conditions

Marine biofouling has severe economic impacts and cyanobacteria play a significant role as early surface colonizers. Despite this fact, cyanobacterial biofilm formation studies in controlled hydrodynamic conditions are scarce. In this work, computational fluid dynamics was used to determine the shear rate field on coupons that were placed inside the wells of agitated 12-well microtiter plates. Biofilm formation by three different cyanobacterial strains was assessed at two different shear rates (4 and 40 s⁻¹) which can be found in natural ecosystems and using different surfaces (glass and perspex). Biofilm formation was higher under low shear conditions, and differences obtained between surfaces were not always statistically significant. The hydrodynamic effect was more noticeable during the biofilm maturation phase rather than during initial cell adhesion and optical coherence tomography showed that different shear rates can affect biofilm architecture. This study is particularly relevant given the cosmopolitan distribution of these cyanobacterial strains and the biofouling potential of these organisms.     

Tectonostratigraphy of Middle and Upper Palaeozoic black shales from the Porto-Tomar-Ferreira do Alentejo shear zone (W Portugal): new perspectives on the Iberian Massif

Middle/late Devonian and early Carboniferous metasedimentary sequences in the northernmost region (Porto-Espinho-Tomar) of the Ossa-Morena Zone (Portuguese Iberian Variscan Massif) contain black shales of very low to low-grade metamorphism. These metasedimentary rocks form a discrete NNW-SSE structure within a major shear zone (Porto-Tomar-Ferreira do Alentejo) and remain subparallel to the observed regional major structures (folding, thrusts or overthrusts). These black shales are overhanged and then imbricated in an upper Proterozoic metamorphic substratum. A multi-disciplinary study of these metasedimentary rocks from the Espinho-Tomar region has tectonostratigraphy, palynology, organic petrology and clay mineralogy combined methods. This approach provides new insights into the tectonic evolution and geological framework of Palaeozoic basement of the Iberian Variscides. Palaeoenvironmental and tectonostratigraphic implications on the Iberian geodynamic framework are discussed.     

Rice Gene <Emphasis Type="Italic">OsDSR-1</Emphasis> Promotes Lateral Root Development in <Emphasis Type="Italic">Arabidopsis</Emphasis> Under High-Potassium Conditions

Rice gene Oryza sativa Drought Stress Response-1 (OsDSR-1) was one of the genes identified to be responsive to drought stress in the panicle of rice at booting and heading stages by both microarray and quantitative real-time PCR analyses. OsDSR-1 encodes a putative calcium-binding protein, and its overexpression in Arabidopsis rendered transgenic plants to produce much shorter lateral roots (LRs) than wild-type (WT) plants in the medium supplemented with abscisic acid (ABA), suggesting that OsDSR-1 may act as a positive regulator during the process of ABA inhibition of LR development. No significant difference was observed in the total LR length between WT and transgenic plants in the media with the increase of only osmotic stress caused by NaCl, LiCl, and mannitol, while transgenic Arabidopsis seedlings appeared to produce larger root systems with longer total LR lengths under high-potassium conditions than WT seedlings. Further analysis showed that external Ca²⁺ was required for the production of larger root systems, indicating that the promotion by OsDSR-1 of the LR development of transgenic Arabidopsis seemed to occur in a Ca²⁺-dependent manner under high-potassium conditions. We propose that OsDSR-1 may function as a calcium sensor of the signal transduction pathway controlling the LR development under high-potassium conditions.     

Denitrification and bacterial community structure in the cascade of six reservoirs on a tropical river in Brazil

Hydrobiologia - Denitrification activity was measured by the acetylene blockage technique in six reservoirs of the Tietê River, southeastern Brazil, in March 1999 and July 1999. The total...     

Early release of eggs and embryos in a brooding ancient asexual ostracod: brood selection or a gambling strategy to increase fecundity?

Asexual lineages lack the means to purge their genomes of (deleterious) mutations through recombination. Evolutionary theory thus predicts that such lineages will be prone to early extinction. In brooding animals, brood selection might provide a mechanism to counter the accumulation of mutations. Of the three putative ancient asexual animal groups, only the darwinulid ostracods are brooders. Here, we test the incidence of egg and juvenile abortion in a darwinulid species, Penthesilenula brasiliensis, under two temperature treatments. Part of the offspring is released without brooding (close to 30% in one treatment). The majority of these aborted eggs hatches and develops. As it is unlikely that females are such bad judges of offspring quality, either the surviving animals will present deficiencies later on in development (brood selection) or early egg release can be a (bet-hedging) strategy to increase fecundity in favourable conditions.     

Disease-causing point-mutations in metal-binding domains of Wilson disease protein decrease stability and increase structural dynamics

After cellular uptake, Copper (Cu) ions are transferred from the chaperone Atox1 to the Wilson disease protein (ATP7B) for incorporation into Cu-dependent enzymes in the secretory pathway. Human ATP7B is a large multi-domain membrane-spanning protein which, in contrast to homologues in other organisms, has six similar cytoplasmic metal-binding domains (MBDs). The reason for multiple MBDs is proposed to be indirect modulation of enzymatic activity and it is thus intriguing that point mutations in MBDs can promote Wilson disease. We here investigated, in vitro and in silico, the biophysical consequences of clinically-observed Wilson disease mutations, G85V in MBD1 and G591D in MBD6, incorporated in domain 4. Because G85 and G591 correspond to a conserved Gly found in all MBDs, we introduced the mutations in the well-characterized MBD4. We found the mutations to dramatically reduce the MBD4 thermal stability, shifting the midpoint temperature of unfolding by more than 20 °C. In contrast to wild type MBD4 and MBD4D, MBD4V adopted a misfolded structure with a large β-sheet content at high temperatures. Molecular dynamic simulations demonstrated that the mutations increased backbone fluctuations that extended throughout the domain. Our findings imply that reduced stability and enhanced dynamics of MBD1 or MBD6 is the origin of ATP7B dysfunction in Wilson disease patients with the G85V or G591D mutation.