Selective degradation of abnormal proteins in mammalian tissue culture cells.

Journal ArticleThe degradation rates of several missense mutants of hypoxanthine-guanine phosphoribosyltransferase (EC 2.4.2.8) in mouse L cells are compared to those of the wild-type enzyme. Although the rates of total protein breakdown in the mutant cell lines are identical to that of the parental L cell line, defective molecules of hypoxanthine-guanine phosphoribosyltransferase present in the mutant cell lines are degraded much faster than the wild-type enzyme. The level of defective phosphoribosyltransferase molecules present in the mutant cell lines is inversely proportional to the breakdown rate. This observation indicates that the major factor determining the concentrations of the defective phosphoribosyltransferases is their specific degradation rate. These results strongly support the hypothesis that abnormal proteins are selectively degraded in mammalian cells

Fundamental cellular processes do not require vertebrate-specific sequences within the TATA-binding protein.

Journal ArticleThe 180-amino acid core of the TATA-binding protein (TBPcore) is conserved from Archae bacteria to man. Vertebrate TBPs contain, in addition, a large and highly conserved N-terminal region that is not found in other phyla. We have generated a line of mice in which the tbp allele is replaced with a version, tbp(Delta N), which lacks 111 of 135 N-terminal amino acid residues. Most tbp(Delta N/Delta N) fetuses die in midgestation. To test whether a disruption of general cellular processes contributed to this fetal loss, primary fibroblast cultures were established from +/+, Delta N/+, and Delta N/Delta N fetuses. The cultures exhibited no genotype-dependent differences in proliferation or in expression of the proliferative markers dihydrofolate reductase (DHFR) mRNA (S phase-specific) and cdc25B mRNA (G(2)-specific). The mutation had no effect on transcription initiation site fidelity by either RNA polymerase II (pol II) or pol III. Moreover, the mutation did not cause differences in levels of U6 RNA, a pol III-dependent component of the splicing machinery, in mRNA splicing efficiency, in expression of housekeeping genes from either TATA-containing or TATA-less promoters, or in global gene expression. Our results indicated that general eukaryotic cell functions are unaffected by deletion of these vertebrate-specific sequences from TBP. Thus, all activities of this polypeptide domain must either be compensated for by redundant activities or be restricted to situations that are not represented by primary fibroblasts

Production of selenoprotein P (Sepp1) by hepatocytes is central to selenium homeostasis

pre-printBackground: Sepp1 transports selenium, but its complete role in selenium homeostasis is not known. Results: Deletion of Sepp1 in hepatocytes increases liver selenium at the expense of other tissues and decreases whole-bodyselenium by increasing excretion. Conclusion: Sepp1 production by hepatocytes retains selenium in the organism and distributes it from the liver to peripheral tissues. Significance: Sepp1 is central to selenium homeostasis

Illegitimate cre-dependent chromosome rearrangements in transgenic mouse spermatids.

Journal ArticleThe bacteriophage P1 Cre/loxP system has become a powerful tool for in vivo manipulation of the genomes of transgenic mice. Although in vitro studies have shown that Cre can catalyze recombination between cryptic "pseudo-loxP" sites in mammalian genomes, to date there have been no reports of loxP-site infidelity in transgenic animals. We produced lines of transgenic mice that use the mouse Protamine 1 (Prm1) gene promoter to express Cre recombinase in postmeiotic spermatids. All male founders and all Cre-bearing male descendents of female founders were sterile; females were unaffected. Sperm counts, sperm motility, and sperm morphology were normal, as was the mating behavior of the transgenic males and the production of two-celled embryos after mating. Mice that expressed similar levels of a derivative transgene that carries an inactive Cre exhibited normal male fertility. Analyses of embryos from matings between sterile Cre-expressing males and wild-type females indicated that Cre-catalyzed chromosome rearrangements in the spermatids that lead to abortive pregnancies with 100% penetrance. Similar Cre-mediated, but loxP-independent, genomic alterations may also occur in somatic tissues that express Cre, but, because of the greater difficulty of assessing deleterious effects of somatic mutations, these may go undetected. This study indicates that, following the use of the Cre/loxP site-specific recombination systems in vivo, it is prudent to eliminate or inactivate the Cre recombinase gene as rapidly as possible

De-excitation spectroscopy of strongly interacting Rydberg gases

We present experimental results on the controlled de-excitation of Rydberg states in a cold gas of Rb atoms. The effect of the van der Waals interactions between the Rydberg atoms is clearly seen in the de-excitation spectrum and dynamics. Our observations are confirmed by numerical simulations. In particular, for off-resonant (facilitated) excitation we find that the de-excitation spectrum reflects the spatial arrangement of the atoms in the quasi one-dimensional geometry of our experiment. We discuss future applications of this technique and implications for detection and controlled dissipation schemes.Comment: 6 pages, 5 figure

A Spatial Multi-criteria Decision Support System for Stress Recovery-Oriented Forest Management

AbstractA solution to cope with chaotic urban settlements and frenetic everyday life is refuging in nature as a way to reduce stress. In general—in recent years—it has been scientifically demonstrated how natural areas are an important environment for psycho-physiological health. As a consequence, it is important to plan dedicated spaces for stress recovery in order to increase the well-being of people. With respect to forests, there is a growing interest in understanding the marketing and tourist potential of forest-therapy activities and policies. This paper develops a decision support system (DSS) for decision makers, based on geographic information system to define the suitability of forest areas to improve psychological and physiological human well-being. Innovative technologies such as electroencephalography (EEG) and virtual reality (VR) are applied to test human status. The DSS combines four sets of indicators in a multi-attribute decision analysis and identifies the areas with the largest stress-recovery potential. Two multi-attribute model—one in summer and one in winter—are elaborated to obtain a dynamic evaluation of suitability. Results show significant differences among forest type, forest management, altitude range, and season in terms of stand suitability. EEG and VR seem to be promising technologies in this research area. Strengths and weaknesses of the approach, as well as potential future improvement and implications for territorial marketing, are suggested

Neuroscience application for the analysis of cultural ecosystem services related to stress relief in forest

The paper presents an integrated methodology to assess psychological and physiological responses of people when exposed to forests, with the main objective of assessing the suitability of different stands for stress recovery on the basis of tree species and density. From the methodological viewpoint, the study applies both a Restoration Outcome Scale (ROS) questionnaire and a neuroscientific technique grounded on electro-encephalographic (EEG) measurement. Results show different outcomes for conifers and broadleaves as well as a statistical significance of density in the evaluation of an individual’s emotional state. A forest with a high density of conifers and low density of broadleaves seems to be the proper combination for stress recovery. The differences among psychological stated preferences and EEG trends highlights potential conflict among “needs” and “wants” of people in the topic of stress relief. Potential applications of the research for health care and territorial marketing operations are suggested