Modification of ColE1 DNA with osmium tetroxide generates positively supercoiled molecules

Covalent binding of osmium tetroxide to negatively supercoiled DNA in vitro initially induces its relaxation, accompanied by a formation of a single denaturation "bubble" per molecule. Binding of further osmium results in DNA overwinding and the appearance of positive supercoils as demonstrated by gel electrophoresis and electron microscopy.     

Differential expression of keratin 19 in normal human epithelial tissues revealed by monospecific monoclonal antibodies

Summary Three monospecific monoclonal antibodies (BA16, BA17 and A53—B/A2) recognizing different epitopes of the human keratin 19 were used to determine tissue distribution of this 40 kDa keratin polypeptide. Immunohistochemical methods revealed four different staining patterns among normal human epithelial tissues: firstly, complete negativity of the epidermis, sebaceous glands, hepatocytes and other tissues; secondly, homogeneous positivity as seen for example in the gall bladder and urinary bladder epithelium, endometrium and many other epithelia; thirdly, a mosaic of positive and negative cells among mammary gland luminal cells, prostate epithelia and some other epithelia and fourthly, a more complex heterogeneous pattern found in non-keratinizing squamous epithelia and hair follicles with generally the basal layer being the most strongly or sometimes exclusively stained. The pattern seen in non-keratinizing squamous epithelia varied considerably according to the fixation method and the antibody used as well as among different donors and in different areas of the same organ. The other three staining patterns were on the other hand nearly identical with all three antibodies on both frozen sections and sections of methacarn-fixed paraffinembedded tissues. Our results provide evidence for differential expression of the human keratin 19 at the single cell level, an observation which could be exploited in the study of epithelial differentiation and pathology.     

Developmentally regulated surface structures of teratocarcinoma stem cells studied by mutant cell lines

Monoclonal antibodies TEC-01, TEC-02, and TEC-03, which define three developmentally regulated antigens TEC-1 (SSEA-1-like), TEC-2, and TEC-3, have been used to isolate and characterize teratocarcinoma stem cell mutants with altered expression of surface glycoconjugates. Mutants lacking TEC-1 antigen have been isolated by exposing mutagenized P19S1801A1 cells to TEC-01 antibody, which was conjugated to the toxin from Ricinus communis. None of the mutants exhibits significant changes in the expression of TEC-3 antigen, but some are defective in the expression of TEC-2 antigen. Analysis of the expression of TEC-1,2,3 antigens in different lectin-resistant F9 and OTF9-63 cell lines has shown that all express TEC-1 antigen, but some lectin-resistant phenotypes exhibit reduction in the expression of TEC-2 and/or TEC-3 antigens. Mutational events in genes regulating the expression of specific glycosyltransferases or glycosidases appear to be the biochemical mechanism regulating the expression of TEC-1 and TEC-2 antigens.     

How does surrounding vegetation affect the course of succession: A five‐year container experiment

Question: How does location and time of insertion affect the course of succession in experimental containers? Location: Benešov nad Lipou, Českomoravská vrchovina (Czech-Moravian uplands), Czech Republic Methods: We designed a 5-year container experiment in which plant succession started from scratch. Soil conditions were constant and all containers were filled with homogeneous substrate containing no propagules. We placed the containers in two contrasting habitats (meadow and floodplain) under identical climatic conditions but differing in surrounding vegetations and hence seed input. New containers were installed (and hence succession started) in two subsequent years, twice in each year (spring and autumn). We assume that the individual dates would lead to differences in propagule input and weather conditions. Results: Although both year and season of succession initiation considerably affected the initial species composition, we observed a pronounced convergence within the set of containers located in each habitat. However, the similarity of containers initiated at the same time but located in different habitats decreased over the course of succession. Final composition of the meadow and floodplain containers was therefore mostly determined by permanent seed input from their nearby neighborhood. Conclusions: This study demonstrated that propagule availability is an important determinant of the course of succession, and that differential seed input leads to different pathways of succession, even when all other environmental conditions are equal.     

Temperate trees and shrubs as global invaders: the relationship between invasiveness and native distribution depends on biological traits

Many woody plants have been recently recognized as major invasive species with serious impacts on species diversity and functioning of invaded ecosystems. We evaluated the global invasion success of temperate trees and shrubs with native ranges in central Europe and explored the role of their native distribution and that of biological traits in determining whether they have become invasive in other parts of the world. Of the 94 species forming the source-area species pool, 27 % are known to be invasive in at least one region of the world. Generalized linear models on phylogenetically non-informed analyses revealed that tall woody plants flowering early in the season, and occupying many grid cells in the native range are significantly more likely to become successful invaders than species not possessing these traits. However, other traits can partly reduce the disadvantages resulting from low regional frequencies in the native range and consequent lower probability of them being introduced elsewhere. Species that do not depend for reproduction on another individual, those pollinated by wind and dispersed by animals are likely to become invasive even if they do not have extensive native distributions. However, of these traits only the regional frequency in the native range remained significant when phylogenetic relationships were taken into account. This indicates that the effect of the regional frequency is generic, valid across all woody species, and fine-tuned by advantageous biological traits inherited from common ancestors, shared by groups of phylogenetically-related species. Traits that only appeared significant in the phylogenetically non-informed analyses should be considered as specific for shrubs and trees of central Europe. Mode of reproduction was only significant in the phylogenetically-informed analysis, indicating that reproduction exclusively by seed favours invasiveness of woody species. From the management perspective, however, the predictive value of some traits is not diminished by them being phylogenetically constrained since we are not interested whether the behaviour of particular species is a result of evolutionary history but how we can treat specific cases of invasion.     

Prey-Specific Growth Responses of Freshwater Flagellate Communities Induced by Morphologically Distinct Bacteria from the Genus Limnohabitans

Because their large growth potential is counterbalanced with grazing by heterotrophic nanoflagellates (HNF), bacteria of the genus Limnohabitans, which are common in many freshwater habitats, represent a valuable model for examining bacterial carbon flow to the grazer food chain. We conducted experiments with natural HNF communities taken from two distinct habitats, the meso-eutrophic Římov Reservoir and the oligo-mesotrophic Lake Cep (South Bohemia). HNF communities from each habitat at distinct seasonal phases, a late April algal bloom and a late May clear water phase, were each fed 3 Limnohabitans strains of differing cell sizes. Water samples were prefiltered (5 μm) to release natural HNF communities from zooplankton control and then amended with the Limnohabitans strains L. planktonicus II-D5 (medium sized, rod shaped), Limnohabitans sp. strain T6-5 (thin, long, curved rod), and Limnohabitans sp. strain 2KL-3 (large solenoid). Using temporal sampling and prey treatment, we determined HNF growth parameters such as doubling time, growth efficiency, and length of lag phase prior starting to exponential growth. All three Limnohabitans strains supported HNF growth but in significant prey-, site-, and season-dependent fashions. For instance, addition of the moderately large T6-5 strain yielded very rapid HNF growth with a short lag phase. In contrast, the curved morphology and larger cell size of strain 2KL-3 made this prey somewhat protected against grazing by smaller HNF, resulting in slower HNF growth and longer lag phases. These trends were particularly pronounced during the late May clear-water phase, which was dominated by smaller HNF cells. This may indicate a longer “adaptation time” for the flagellate communities toward the large prey size offered.     

Chlorophyll-phosphorus relationship in acidified lakes of the High Tatra Mountains (Slovakia)

Concentrations of total phosphorus (TP) and chlorophyll a (CHLA) were measured in 28 lakes in the High Tatra Mountains (Slovakia) from 1983 to 1990. The relationship between log CHLA and log TP in the Tatra lakes is similar to relationships developed for lakes in other regions, but variation is higher. A part of this variation is caused by acidification of the lakes. In the lakes with pH between 4.9 and 6.3 the CHLA concentrations are often extremely low while TP concentrations decreased, but not as drastically.