Effect of enterococcin A 2000 on biological and synthetic phospholipid membranes

Lactic acid bacterium isolated from Bulgarian cheese and identified as Enterococcus faecium produces a small hydrophobic peptide substance (enterococcin A 2000) with broad spectrum of antimicrobial activity. The wide range of enterococcin antibacterial activity of this compound against Gram-positive, as well as against some Gram-negative bacteria, suggests a single mechanism of action. The mode of action of enterococcin A 2000 was studied in intact liver mitochondria and synthetic phospholipid liposomes used as model systems. Enterococcin A 2000 stimulated the ATPase activity in intact mitochondria. The kinetic curve of ATP hydrolysis differed from that obtained in presence of dinitrophenol (DNP) and showed a character similar to the ATP hydrolysis in the presence of classic ionophores. Enterococcin A 2000, when bound to synthetic phospholipid liposomes, permeabilized liposomes liberating the marker carboxyfluorescein (CF).     

Structural characterization of chromosome I size variants from a natural yeast strain

Many yeast strains isolated from the wild show karyotype instability during vegetative growth, with rearrangement rates of up to 10(-2) chromosomal changes per generation. Physical isolation and analysis of several chromosome I size variants of one of these strains revealed that they differed only in their subtelomeric regions, leaving the central 150 Kb unaltered. Fine mapping of these subtelomeric variable regions revealed gross alterations of two very similar loci, FLO1 and FLO9. These loci are located on the right and left arms, respectively, of chromosome I and encompass internal repetitive DNA sequences. Furthermore, some chromosome I variants lacking the FLO1 locus showed evidence of recombination at a DNA region on their right arm that is enriched in repeated sequences, including Ty LTRs. We propose that repetitive sequences in many subtelomeric regions in S. cerevisiae play a key role in karyotype hypervariability. As these regions encode several membrane-associated proteins, subtelomeric plasticity may allow rapid adaptive changes of the yeast strain to specific substrates. This pattern of semi-conservative chromosomal rearrangement may have profound implications, both in terms of evolution of wild strains and for biotechnological processes.     

Variation in ovarian morphology in four species of New World moles with a peniform clitoris

Female moles of the Old World genus Talpa display a curious suite of reproductive features that include a peniform clitoris and ovaries with a discrete interstitial gland or testis-like region (so-called 'ovotestes'). The masculinization of the female external genitalia in Talpa has accordingly been linked with secretion of androgens from the interstitial gland region of the fetal gonad. Although their ovarian morphology has received less attention, some species of New World moles also have ovaries with a pronounced interstitial gland (for example star-nosed mole, Condylura cristata), whereas females of other species do not (for example eastern mole, Scalopus aquaticus). Although it is difficult to determine the sex of both Old and New World moles, published accounts describing the external genitalia of female moles are available only for Talpa. The hypothesis that masculinization of the female external genitalia in moles is associated with the presence of an ovarian interstitial gland (OIG) was tested in the present study by using a comparative approach to determine whether these features are ever found in isolation of one another. Three genera of North American moles (Scapanus, Condylura and Neurotrichus) were studied and a peniform clitoris was found in all three species, but OIG were found in only two of three genera. The ovaries of S. latimanus and S. orarius were unremarkable, with no evidence of a discrete interstitial gland or testis-like region. Mapping these results onto recent talpid phylogenies indicates that loss of the bipolar ovarian morphology is a derived trait in Scapanus, and conclusively demonstrates that masculinization of the external genitalia in female moles can develop in the presence or absence of 'ovotestes'.     

Immunocytochemical study of alpha 1 and beta 2/3 subunits of GABAA receptors in freehand isolated vestibular Deiters' neurons

Vestibular Deiters' neurons have been isolated from bovine brain by the Hydén's freehand dissection technique and challenged with monoclonal antibodies directed toward the alpha 1 and beta 2/3 subunits of the GABAA receptors. Subsequent challenge with fluorescent secondary antibodies and confocal microscopy allowed the study of the cellular distribution of such subunits. In Deiters' neurons the beta 2/3 subunit displayed a clear presence all along the cell body profile and the initial parts of the dendrites. The alpha 1 subunit was found highly present all over the cell interior except the nuclear profiles. The strong presence inside the cells possibly masked its presence on the plasma membrane. However, in part of the cells studied a distinct presence on the plasma membrane was evident. This subunit was visualized also all along the long dendrites of these neurons. The approach we describe here, involving freehand isolated mature neurons from adult animals, may allow a better characterization of the tridimensional distribution of different types of neuronal GABAA receptors in the respect of the approach with brain slices.     

The biomass dynamics of cyanobacteria in an annual cycle determined by confocal laser scanning microscopy

We recently published a method for estimating cyanobacteria biomass in delta microbial mats from the Ebro river using confocal laser scanning microscopy (CLSM). The present paper uses this method for identifying different groups of cyanobacteria and for determining their biomass dynamics. Microcoleus chthonoplastes and the Lyngbya-Oscillatoria were the most important contributors to the cyanobacterial biomass throughout the study. Biomass values ranged from 1.29 to 6.55 mgC/cm2 sediment for Microcoleus chthonoplastes, and from 128 microgC/cm2 to 3.16 mgC/cm2 sediment for Lyngbya-Oscillatoria. This technique is useful for determining biomass and for studying filamentous cyanobacteria as well as ramified eukaryotic cells. Confocal serial sections through the samples can be obtained. Two-dimensional images from the samples can be used to calculate the biomass of individual cells.     

Building world city Tokyo: Globalization and conflict over urban space

Japanese policy makers have, since their contact with the colonial powers in the mid 19th century, been acutely aware of the pressures and challenges of national survival in a globalizing world. In this sense, the Japanese experience of modernity has been deeply intertwined with, and is in important ways inseparable from the ongoing processes of globalization during the last century and a half. While their main response was to foster the growth of Japanese industrial, military and diplomatic power, one consistent theme has been the development of the capital city Tokyo as emblem of Japan as a civilized nation, location of national institutions, and center of economic power. This project, however, has long been an arena of considerable conflict between city builders and the residents of central Tokyo. The most recent conflict over the control of urban space in Japan's premier world city emerged in the last few years when major developers lobbied successfully for massive increases in allowable building volumes and heights in special regeneration areas, arguing that without further deregulation Tokyo would lose its competitive position in relation to Shanghai, Hong Kong and Singapore. This paper argues that in Japan an important feature of globalization and international competitive pressures has been their use by urban actors in disputes over the control of urban space, and examines this use of globalization debates in the competition between economic space and life space in Tokyo.     

NORM workers: a challenge for internal dosimetry programmes

The mining, milling and processing of uranium and thorium bearing minerals may result in radiation doses to workers. The control of occupational exposures from these natural sources of radiation imposes a challenge to regulators and radiation protection advisers. A survey pilot programme, which included six mines in Brazil and a monazite plant, was established, consisting of the collection and analysis of concentrations of uranium, thorium and polonium in urine, faeces and air samples. Results from workers were compared to background data from their families living in the same area and from residents from the population of Rio de Janeiro. Positive exposure results were identified among the coal miners, the niobium miners and the monazite sand workers. Difficulties in the application of internal dosimetry programmes are discussed in relation to the control of NORM workers.     

Simulation of multistage extraction of antioxidants from Chilean hazelnut (<Emphasis Type="Italic">Gevuina avellana</Emphasis>) hulls

The objective of this study was to analyze the factors affecting the ethanol extraction of Chilean hazelnut (Gevuina avellana) hulls to obtain antioxidant compounds. The effects of temperature on the kinetics of polyphenolics extraction and on the antioxidant activities of the extracts were assessed. The radical-scavenging activities of the extracts were comparable with that of BHA when used at the same concentration. The optimal temperature for the antioxidant activities of the extracts was 40°C. A four-stage cross-flow extraction was carried out and a four-stage countercurrent extraction was simulated, where each stage lasted 30 min. Best results were obtained with countercurrent extraction, which produced an extract that showed 94.4% α,α-diphenyl-β-picrylhydazyl radical inhibition, compared with 86.2% obtained in the first stage of cross-flow extraction and 92.62% in a batch extraction that lasted 100 h. UV-vis and NIR spectra of extracts from cross-flow and from the simulated countercurrent extraction revealed that the composition of extracts varied along the stages and was affected by the operational strategy.