Revertants of Adenovirus Type 12-Transformed Hamster Cell Line T637 as Tools in the Analysis of Integration Patterns

Spontaneously arising morphological revertants of the adenovirus type 12 (Ad12)-transformed hamster cell line T637 had been previously isolated, and it had been demonstrated that in these revertants varying amounts of the integrated Ad12 genome were eliminated from the host genome. In this report, the patterns of persistence of the viral genome in the revertants were analyzed in detail. In some of the revertant cell lines, F10, TR3, and TR7, all copies of Ad12 DNA integrated in line T637 were lost. In lines TR1, -2, -4 to -6, -8 to -10, and -13 to -16, only the right-hand portion of one Ad12 genome was preserved; it consisted of the intact right segment of Ad12 DNA and was integrated at the same site as in line T637. In revertant lines G12, TR11, and TR12, one Ad12 DNA and varying parts of a second viral DNA molecule persisted in the host genome. These patterns of persistence of Ad12 DNA molecules in different revertants supported a model for a mode of integration of Ad12 DNA in T637 hamster cells in which multiple (20 to 22) copies of the entire Ad12 DNA were serially arranged, separated from each other by stretches of cellular DNA. The occurrence of such revertants demonstrated that foreign DNA sequences could not only be acquired but could also be lost from eucaryotic genomes. There was very little, if any, expression of Ad12-specific DNA sequences in the revertant lines TR7 and TR12. Moreover, Ad12 DNA sequences which were found to be undermethylated in line T637 were completely methylated in the revertant cell lines G12, TR11, TR12, and TR2. These findings were consistent with the absence of T antigen from the revertant lines reported earlier. Hence it was conceivable that the expression of integrated viral DNA sequences was somehow dependent on their positions in the cellular genome. In cell line TR637, the early segments of Ad12 DNA were expressed and undermethylated; conversely, in the revertant lines G12, TR11, TR12, and TR2, the same segments appeared to be expressed to a limited extent and were strongly methylated.     

Microphotometric measurement of initial maximum reaction rates in quantitative enzyme histochemistryin situ

Summary Final reaction product formation was recorded microphotometrically for succinate dehydrogenase in cross-sectioned muscle fibres at initial rate conditions and during prolonged incubations. Incubations with gel films and aqueous reaction medium both showed a decline of reaction rates. Maximum reaction rates could only be determined at initial rate conditions during the first minute of the incubation. Reaction rates recorded in different areas of the same tissue section were found to change with time to different degrees. From these results it was concluded that quantitative histochemical measurements of enzyme reactionsin situ can only be valid if measured under initial maximum velocity conditions.     

Les otolithes des Téléostéens du Plio-Pleistocène belge

The study of approximately twenty three thousand otoliths from Plio-Pleistocene sites in the harbour region of Antwerp, as well as a critical revision of already published material allowed us to identify a teleostean fauna with fourty seven species (including two subspecies and seven species in open nomenclature). Two of these, Ophidion springeri and Uranoscopus septentrionalis are new to science. The fauna is typical for coastal waters slightly warmer than those of the actual North Sea and caracterized by the predominance of Gadidae. Biostratigraphically this fauna is well individualized with respect to preceeding ones (only seven species in common with the Miocene fauna) and those following twenty-three species in common with the extant fauna). The following biostratigraphical subdivision has been recognized in the Upper Miocene and the Plio-Pleistocene of the Antwerp region: (1) an association with «genus? Macrouridarum labiatus, Trisopterus sculptus, Gadiculus benedeni and Trisopterus luscus spectabilis, in the Sands of Deurne (Upper Miocene); (2) an association with Gadiculus benedeni and Merlangius pseudaeglephinus, in the Formation of Kattendijk (Pliocene); (3) an association with Gadiculus verticalis and Merlangius pseudaeglefinus in the Formation of Lillo (Plio-Pleistocene); (4) an association with Gadus morhua and Merlangius pseudaeglefinus in the so called «Icenian deposits not present in Belgium but found in the Netherlands.     

Observations nouvelles sur les otolithes des Téléostéens du Calcaire Grossier (Eocène du bassin de Paris)

An important new collection of otoliths from theCalcaire Grossier of the Paris Basin, as well as a critical review of already published material is studied and reveals the presence of 56 species. Five of these are new: Muraenesox spatulus, Genus Synodontidarum intermedius, Genus Ophidiidarum spinosus, Chanda bohlkei and Mene sekharani. The assemblage is indicative of tropical to subtropical marine and neritic environment with solid bottom. Biogeographically, it shows affinities with the recent Indo-pacific fauna. The association of the sites at Fercourt and Château-Rouge is clearly different from the one found at the sites of Condé-en-Brie and Damery, as it is less littoral than the second association mentioned.     

Molecular Co-occupancy Identifies Transcription Factor Binding Cooperativity In Vivo

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Dark diversity in Amazonian stream fish communities: What factors determine species absence along environmental gradients?

1. Species distribution models often fail to predict observed patterns of species diversity, and this is because some species within a regional pool that are tolerant of conditions at a given location may nevertheless be absent from the local community. These missing species have been termed “dark diversity”. In the present study, we investigated which factors explain dark diversity among fish assemblages in Amazonian streams.
2. We sampled 71 streams in areas with different types of land use within two river basins and estimated dark diversity from patterns of species co-occurrence, using Beals’ index, along environmental gradients. From this procedure, taxa are designated as dark diversity components when they are absent from a given stream, but often co-occur with the local species at other streams, indicating similar ecological requirements. We used generalised linear models both to determine whether environmental or landscape variables, connectivity, instream environmental heterogeneity or some combination of these factors explained dark diversity of fishes, and to evaluate whether ecomorphology is associated with the extent to which a species contributes to dark diversity and which specific traits contribute the most to explaining variation in dark diversity.
3. Mean local diversity exceeded observed dark diversity. The magnitude of dark diversity was directly associated with the proportion of secondary forest in the immediate catchment and with the index of proximity to anthropogenic impact. Species that have high affinity for environments with higher current velocity, low swimming ability and that capture food mainly on the surface contributed more to dark diversity, which suggests that swimming ability, habitat preference and aspects related to diet are key predictors of the probability that a given species will be present at locations with suitable habitat.
4. Our findings reinforce the idea that dark diversity results from interactions between species traits and environmental factors, including anthropogenic impacts. Understanding the interplay among environmental factors and species traits that contribute to dark diversity provides targets for improved ecosystem restoration and sustainability of native species assemblages.

     

Development,characterization, and application of a 2‐Compartment system to investigate the impact of pH inhomogeneities in large‐scale CHO‐based processes

Large‐scale bioreactors for the production of monoclonal antibodies reach volumes of up to 25 000 L. With increasing bioreactor size, mixing is however affected negatively, resulting in the formation of gradients throughout the reactor. These gradients can adversely affect process performance at large scale. Since mammalian cells are sensitive to changes in pH, this study investigated the effects of pH gradients on process performance. A 2‐Compartment System was established for this purpose to expose only a fraction of the cell population to pH excursions and thereby mimicking a large‐scale bioreactor. Cells were exposed to repeated pH amplitudes of 0.4 units (pH 7.3), which resulted in decreased viable cell counts, as well as the inhibition of the lactate metabolic shift. These effects were furthermore accompanied by increased absolute lactate levels. Continuous assessment of molecular attributes of the expressed target protein revealed that subunit assembly or N‐glycosylation patterns were only slightly influenced by the pH excursions. The exposure of more cells to the same pH amplitudes further impaired process performance, indicating this is an important factor, which influences the impact of pH inhomogeneity. This knowledge can aid in the design of pH control strategies to minimize the effects of pH inhomogeneity in large‐scale bioreactors.