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Cdc13 is a single stranded telomere binding protein that specifically localizes to the telomere ends of budding yeasts and is essential for cell viability. It caps the ends of chromosomes thus preventing...Full Text Available

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Yeast Sir2 deacetylase is a component of the silent information regulator (SIR) complex encompassing Sir2/Sir3/Sir4. Sir2 is recruited to telomeres through Rap1, and this complex spreads into subtelomeric...Full Text Available

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Transcriptional silencing in Saccharomyces cerevisiae occurs at several genetic loci, including the ribosomal DNA (rDNA). Silencing at telomeres (telomere position effect [TPE])...Full Text Available

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DNA replication programs have been studied extensively in yeast and animal systems, where they have been shown to correlate with gene expression and certain epigenetic modifications. Despite the conservation...Full Text Available

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We have investigated the structures formed by oligonucleotides composed of two or four repeats of the telomeric sequences from Oxytricha and Tetrahymena. The Oxytricha four-repeat molecule (d(T4G4)4 = Oxy-4) forms structures with increased electrophoretic mobility in nondenaturing gels containing Na+, K+, or Cs+, but not in gels containing Li+ or no added salt. Formation of the folded structure results in protection of a set of dG's from methylation by dimethyl sulfate. Efficient UV-induced cross-links are observed in Oxy-4 and the related sequence from Tetrahymena (d(T2G4)4 = Tet-4), and join thymidine residues in different repeats. Models proposed to account for these data involve G-quartets, hydrogen-bonded structures formed from four guanosine residues in a square-planar array. We propose that the G-quartet structure must be dealt with in vivo by the telomere replication machinery.

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Lethal alleles of the Drosophila k43 gene result in small or missing imaginal discs, greatly reduced mitotic index, and fragmented and abnormally condensed chromosomes. A female-sterile...Full Text Available

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The goal of the present work was to determine the impact of N3-methyladenine (3-mA), an important lesion generated by many environmental agents and anticancer drugs, on in vivo DNA replication and in vitro RNA transcription. Due to 3-mA chemical instability, the stable isostere 3-methyl-3-deazaadenine (3-m-c(3)A) was site specifically positioned into an oligodeoxynucleotide. The oligomer was, then incorporated into a vector system that is rapidly converted to ssDNA inside yeast cells and requires DNA replication opposite the lesion for plasmid clonal selection. For control purposes, an adenine or a stable apurinic/apyrimidinic (AP)-lesion was placed at the same site. The presence of each lesion in the oligonucleotide was confirmed by MALDI-TOF analysis. Plasmids were then transfected into yeast cells. While the AP-site dramatically reduced plasmid replication in all strains, the ...

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The processes regulating telomere function have major impacts on fundamental issues in human cancer biology. First, active telomere maintenance is almost always required for full oncogenic transformation...Full Text Available

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BackgroundLeukocyte telomere length (LTL) is an emerging marker of biological age. Chronic inflammatory activity is commonly proposed as a promoter of ...Full Text Available

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Myriad genetic and epigenetic alterations are required to drive normal cells toward malignant transformation. These somatic events commandeer many signaling pathways that cooperate to endow aspiring...Full Text Available

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ABSTRACT. Chlorarachniophytes are enigmatic marine unicellular algae that acquired photosynthesis by secondary endosymbiosis. Chlorarachniophytes are unusual in that the nucleus of the engulfed algal cell (a green alga) persists in a miniaturized form, termed a nucleomorph. The nucleomorph genome of the model chlorarachniophyte, Bigelowiella natans CCMP621, is 373 kilobase pairs (kbp) in size, the smallest nuclear genome characterized to date. The B. natans nucleomorph genome is composed of three chromosomes, each with canonical eukaryotic telomeres and sub telomeric ribosomal DNA (rDNA) operons transcribed away from the chromosome end. Here we present the complete rDNA operon and telomeric region from the nucleomorph genome of Lotharella oceanica CCMP622, a newly characterized chlorarachn...

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Heterochromatin constitutes a significant portion of the genome in higher eukaryotes; approximately 30% in Drosophila and human. Heterochromatin contains a high repeat DNA content and a low density of protein-encoding genes. In contrast, euchromatin is composed mostly of unique sequences and contains the majority of single-copy genes. Genetic and cytological studies demonstrated that heterochromatin exhibits regulatory roles in chromosome organization, centromere function and telomere protection. As an epigenetically regulated structure, heterochromatin formation is not defined by any DNA sequence consensus. Heterochromatin is characterized by its association with nucleosomes containing methylated-lysine 9 of histone H3 (H3K9me), heterochromatin protein 1 (HP1) that binds H3K9me, and Su(var)3-9, which methylates H3K9 and binds HP1. Heterochromatin formation and functions are influenced by HP1, Su(var)3-9, and the RNA interference (RNAi) pathway. My thesis project ...

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A mesophilic yeast, Candida utilis, and a psychrophilic yeast, Leucosporidium stokesii, were subjected to freeze-thaw cycling over the range 25 to -60 C. Viability...Full Text Available

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AIM: To analyze the antiviral mechanism of Epigallocatechin gallate (EGCG) against hepatitis B virus (HBV) replication.METHODS: In this research, the HBV-replicating cell line HepG2.117 was used...Full Text Available

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We explored transcriptional responses of the fission yeast Schizosaccharomyces pombe to various environmental stresses. DNA microarrays were used to characterize changes in expression...Full Text Available

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Physical activity is associated with reduced risk of several age-related diseases as well as with increased longevity in both rodents and humans. Though these associations are well established, evidence...Full Text Available

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The COOH-terminus of telomerase reverse transcriptase (hTERT) has been shown to participatein the nuclear translocation of TERT. Here, we constructed plasmids expressing the COOH-terminal M(r) 27,000 polypeptide of hTERT (hTERTC27) withthe telomerase RNA-binding domains and the reverse transcriptase domains deleted. We showed that ectopic overexpression of this polypeptide caused a defect in telomere maintenance in hTERT-positive HeLa cells, which led to senescence-like growth arrest and apoptosis. The hTERTC27 appears to work by inducing telomere dysfunction, exemplified by significantly increased anaphase chromosome end-to-end fusion events in transfected cells. Significantly, it had no effect on the cellular telomerase enzymatic activity or telomere length. The in vivo effect was further demonstrated as HeLa cells stably expressing hTERTC27 have significantly lower growth rate and reduced tumorigenicity in nude mice ...

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de Vito, Peter C. (Princeton University, Princeton, N.J.), and Jacques Dreyfuss. Metabolic regulation of adenosine triphosphate sulfurylase in yeast. J. Bacteriol. 88:1341–1348....Full Text Available

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The electrokinetic patterns of four bacterial species (Bacillus subtilis, Bacillus megaterium, Pseudomonas aeruginosa, and Agrobacterium radiobacter), two yeasts (Saccharomyces cerevisiae and Candida...Full Text Available

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BackgroundHistone post-translational modifications are critical for gene expression and cell viability. A broad spectrum of histone lysine residues have been identified in yeast...Full Text Available

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... e Biologia Molecular (SBBq), Sao Paulo, SP (Brazil) 217 p. APPLIED LIFE

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The lesion bypass pathway, which is regulated by monoubiquitination of proliferating cell nuclear antigen (PCNA), is essential for resolving replication stalling due to DNA lesions. This process is...Full Text Available

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Human cytomegalovirus (HCMV) in clinical material cannot replicate efficiently in vitro until it has adapted by mutation. Consequently, wild-type HCMV differ fundamentally from the passaged strains...Full Text Available

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BackgroundReplication capacity (RC) of specific HIV isolates is occasionally blamed for unexpected treatment responses. However, the role of viral RC in response to antiretroviral...Full Text Available

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Chromosomal common fragile sites (CFSs) are genetically unstable regions of the genome that are induced by conditions that impair DNA replication. In this report, we show that treatment with the DNA...Full Text Available

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CDC7 is an essential gene required for DNA replication in Saccharomyces cerevisiae. Cdc7p homologs have recently been identified in vertebrates, but their role in DNA...Full Text Available

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The genomic termini of RNA viruses contain essential cis-acting signals for such diverse functions as packaging, genome translation, mRNA transcription, and RNA replication, and thus...Full Text Available

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... i3a. ... gives performance measurements for various benchmarks, and compares the performance of Harp with the standard implementation of NFS in ...

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Escherichia coli rnh mutants lacking RNase H activity are capable of recA+-dependent DNA replication in the absence of concomitant protein synthesis (stable DNA replication). In rnh dnaA::Tn10 and rnh...Full Text Available

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Physiological functions are carried out by differentiated cells, with finite lifespans, which age and need to be replaced. In young individuals, tissue functions are sustained at optimal levels because cellular dysfunction and cell loss are balanced by the emergence of newly differentiated cells as stem cells and their partially differentiated descendants replicate. However, with the passage of time the mitotic rates of these cells diminish. Eventually, replications occur too infrequently to offset the loss. It is at this point that the tissue begins to show structural changes and declining function which, as they become pervasive, are identified as ageing. In this paper the theory is set forth that: (1) Diminishing mitotic activity in older tissues results from limited stem cell replicative capacity. (2) All stem cells, regardless of tissue, exhibit similar replicative patterns over time, progressing ...

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The role of intermediate products of dioxygen reduction in cytotoxic effects ascribed to oxygen molecules was studied in vivo using various yeast mutants with changed response to oxygen stress. It has been documented that superoxide radical exerts its deleterious effects on yeast cells directly and the role of other oxygen species derived from it is hardly detectable. Yeast Saccharomyces cerevisiae, however, cannot be considered as a typical eukaryotic organism due to its inability of synthesizing polyunsaturated fatty acids (pufa). These fatty acids are known as main target molecules during oxidative stress and their peroxidation leads to cytotoxic effects. As fatty acid content could be easily manipulated in yeast, this organism was used to evaluate the contribution of pufa peroxidation process to the cytotoxic effects of oxygen. Results obtained show, that yeast cells containing ...

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1. A high degree of homology in the positions of tyrosine residues in glyceraldehyde 3-phosphate dehydrogenase from lobster and pig muscle, and from yeast, prompted an examination of the reactivity...Full Text Available

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A compliant cantilevered three-dimensional micromold is provided. The compliant cantilevered micromold is suitable for use in the replication of cantilevered microparts and greatly simplifies the replication of such cantilevered parts. The compliant cantilevered micromold may be used to fabricate microparts using casting or electroforming techniques. When the compliant micromold is used to fabricate electroformed cantilevered parts, the micromold will also comprise an electrically conducting base formed by a porous metal substrate that is embedded within the compliant cantilevered micromold. Methods for fabricating the compliant cantilevered micromold as well as methods of replicating cantilevered microparts using the compliant cantilevered micromold are also provided.

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While autophagy has been shown to act as an anti-viral defense, the Picornaviridae avoid and, in many cases, subvert this pathway to promote their own replication. Evidence indicates...Full Text Available

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We report that the fission yeast nucleoporin Nup124p is required for the nuclear import of both, retrotransposon Tf1-Gag as well as the retroviral HIV-1 Vpr. Failure to import Tf1-Gag into the nucleus...Full Text Available

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The yeast phosphatidylinositol transfer protein (Sec14p) is required for biogenesis of Golgi-derived transport vesicles and cell viability, and this essential Sec14p requirement is abrogated by inactivation...Full Text Available

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Fission yeast S. pombe is assumed to be a good model for cloning of human DNA repair genes, because human gene is normally expressed in S. pombe and has a very similar protein sequence to yeast protein. We have tried to elucidate the DNA repair mechanisms of S. pombe as a model system for those of mammals. (J.P.N.)

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Histone-beta-galactosidase protein fusions were used to identify the domain of yeast histone 2B, which targets this protein to the nucleus. Amino acids 28 to 33 in H2B were required for nuclear localization...Full Text Available

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Suicide substrate β, γ-bidentate Rh(III)ATP (RhATP) was used to map the metal ion-binding site in yeast phosphoglycerate kinase (PGK). Cleavage of the RhATP-inactivated enzyme with...Full Text Available

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Transcription for a hybrid SV40 promoter-beta globin coding sequence recombinant initiates from both early-early (EE) and late-early (LE) SV40 start sites (EES and LES) in the absence of DNA replication....Full Text Available

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We report the existence of a sixth replication arrest site, TerF, that is located within the coding sequences of the rcsC gene, a negative regulator of capsule biosynthesis. The TerF site is oriented...Full Text Available

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Over the decades, basic research in life sciences has profited greatly from the study of the small unicellular fungal species Saccharomyces cerevisiae. This yeast turned out to be key for the identification and understanding of molecular mechanisms that underlay the basic functions of all eukaryotic cells. These include, but are not limited to, the regulatory mechanisms behind cellular reproduction (cell cycle control), cellular morphogenesis (cell polarity, cytoskeleton and membrane trafficking) and the management of cellular information (chromosome biology, transcription and translation). Rapid access to genomic information of many yeast species, combined with bioinformatics analyses, provide information on the evolutionary history of yeasts and the molecular ancestry of their constituen...

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Abstract Human mesenchymal stem cells (hMSCs) may be used for therapeutic applications. Culture conditions such as the serum source may impact on cell quality and the onset of replicative senescence. We have examined the effect of culturing hMSCs in autologous serum (AS) versus fetal bovine serum (FBS) on factors involved in in vitro replicative senescence. hMSCs from four donors were cultured in 10% FBS or 10% AS until they reached senescence. Cells were harvested at early passage and near senescence to study factors known to be involved in cellular senescence. The number of population doublings till senescence was similar for cells cultured in FBS, but varied greatly for hMSCs cultured in AS. FBS cells accumulated in S phase of cell cycle. This could not be explained by increased express...

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Tropomyosin (Tm) is a conserved dimeric coiled-coil protein, which forms polymers that curl around actin filaments in order to regulate actomyosin function. Acetylation of the Tm N-terminal...Full Text Available

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The exocyst is an evolutionarily conserved octameric complex involved in polarized exocytosis from yeast to humans. The Sec3 subunit of the exocyst acts as a spatial landmark for exocytosis through...Full Text Available

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Eukaryotic cells can massively transport their own cytoplasmic contents into a lytic compartment, the vacuole/lysosome, for recycling through a conserved system called autophagy. The key process in...Full Text Available

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Mutant strains of the yeast Saccharomyces cerevisiae which lack functional Cu,Zn superoxide dismutase (SOD-1) do not grow aerobically unless supplemented with methionine. The molecular basis of this...Full Text Available

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To gain insight into the process of mitochondrial transmission in yeast, we directly labeled mitochondrial proteins and mitochondrial DNA (mtDNA) and observed their fate after the fusion of two cells....Full Text Available

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SummaryThe general transcription factor TFIID is a large multi-subunit complex required for the transcription of most protein-encoding genes by RNA polymerase II. Taking advantage...Full Text Available

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Baker's yeast (Saccharomyces cerevisiae) has been genetically engineered to ferment the pentose sugar xylose present in lignocellulose biomass. One of the reactions controlling the...Full Text Available

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Calorie restriction (CR) induces a metabolic shift towards mitochondrial respiration; however, molecular mechanisms underlying CR remain unclear. Recent studies suggest that CR-induced mitochondrial...Full Text Available

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A fuzzy-decision-making procedure is applied to find the optimal feed policy of a fed-batch fermentation process for fuel ethanol production using a genetically engineered Saccharomyces yeast 1400 (pLNH33). The policy consisted of feed flow rate, feed concentration, and fermentation time. The recombinant yeast 1400 (pLNH33) can utilize glucose and xylose simultaneously to produce ethanol. However, the parent yeast utilizes glucose only. A partially selective model is used to describe the kinetic behavior of the process. In this study, this partially selective fermentation process is formulated as a general multiple-objective optimal control problem. By using an assigned membership function for each of the objectives, the general multiple-objective optimization problem can be converted into a maximizing decision problem. In order to obtain a global solution, a hybrid method of differential evolution is introduced to solve ...

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Spinocerebellar ataxia 10 (SCA10) is an autosomal dominant disease caused by large-scale expansions of the (ATTCT)_n repeat within an intron of the human ATXN10 gene. In contrast...Full Text Available

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The programming of cellular networks to achieve new biological functions depends on the development of genetic tools that link the presence of a molecular signal to gene-regulatory activity. Recently,...Full Text Available

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A high-throughput assay for enzyme activity has been developed that is reaction independent. In this assay, a small-molecule yeast three-hybrid system is used to link enzyme catalysis to transcription...Full Text Available

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BackgroundThe production of high yields of recombinant proteins is an enduring bottleneck in the post-genomic sciences that has yet to be addressed in a truly rational manner. Typically...Full Text Available

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In the yeast Saccharomyces cerevisiae, a and α mating-type information is stored in transcriptionally silenced cassettes called HML and HMR....Full Text Available

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Cultured fish and human cells have been used as bioassay systems for the evaluation of the toxicity of aquatic pollutants. Numerous assays using bacteria and yeast have also been used for such purposes. The authors report the toxicity of aquatic pollutants (Cd, Hg, and Ni), using cell culture systems and the yeast Saccharomyces cervisiae test. Cd, Hg, and Ni were chosen as model compounds of pollutants because the related toxicity is now fairly well established.

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Cytosine methylation in DNA is a major epigenetic signal, and plays a central role in propagating chromatin status during cell division. However the mechanistic links between DNA methylation and histone methylation are poorly understood. A multi-domain protein UHRF1 (ubiquitin-like, containing PHD and RING finger domains 1) is required for DNA CpG maintenance methylation at replication forks, and mouse UHRF1-null cells show enhanced susceptibility to DNA replication arrest and DNA damaging agents. Recent data demonstrated that the SET and RING associated (SRA) domain of UHRF1 binds hemimethylated CpG and flips 5-methylcytosine out of the DNA helix, whereas its tandom tudor domain and PHD domain bind the tail of histone H3 in a highly methylation sensitive manner. We hypothesize that UHRF1 brings the two components (histones and DNA) carrying appropriate markers (on the tails of H3 and hemimethylated CpG sites) ready to be assembled into a ...

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Replication of plus-strand RNA viruses depends on host factors that are recruited into viral replicase complexes. Previous studies showed that eukaryotic translation elongation factor (eEF1A) is one...Full Text Available

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Virulent strains of Newcastle disease virus ([NDV] also known as avian paramyxovirus type 1) can be discriminated from low-virulence strains by the presence of multiple basic amino acid residues at...Full Text Available

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BackgroundAzithromycin (AZM) is a macrolide antibiotic that displays an excellent safety profile even in children and pregnant women and has been shown to have anti-malarial activity...Full Text Available

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Human chromosomal fragile sites are specific loci that are especially susceptible to DNA breakage following conditions of partial replication stress. They often are found in genes involved in tumorigenesis...Full Text Available

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We previously reported that global deletion of insulin receptor substrate protein 1 (Irs1) extends lifespan and increases resistance to several age-related pathologies in female mice....Full Text Available

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The HTLV-1 Tax protein both activates viral replication and is involved in HTLV-1-mediated transformation of T lymphocytes. The transforming properties of Tax include altering the expression of select...Full Text Available

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The identification of host cell factors for virus replication holds great promise for the development of new anti-viral therapies. Recently, high-throughput screening methods have emerged as...Full Text Available

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SummaryDnaA initiates chromosomal replication in E. coli at a well regulated time in the cell-cycle. To determine how the spatial distribution of DnaA is related...Full Text Available

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Generation of replicative defective viruses is frequently observed during viral infections. We now report that encapsidation and reverse transcription of spliced viral RNA is an additional mechanism...Full Text Available

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We recently reported that human immunodeficiency virus type 1 (HIV-1) unintegrated linear DNA displays a discontinuity in its plus strand, precisely defined by a second copy of the polypurine tract...Full Text Available

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Six patient assessment systems that have explicit decision rules for replicating team judgments on level of care patient placement were selected for analysis. The six were selected because of their...Full Text Available

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The principle aim of the program was to produce a novel, non-leaching antimicrobial surface for commercial development and future use in the liquid food packaging industry. Antimicrobial surfaces which exist presently have been produced to combat the growth of prokaryotic organisms and usually function as slow release systems. A system which could inhibit eukaryotic growth without contaminating the surrounding 'environment' with the inhibitor was considered of great commercial importance. The remit of this study was concerned with creating a surface which could control the growth of eukaryotic organisms found in fruit juice with particular interest in the yeast, Saccharomyces cerevisiae. Putative antimicrobial surfaces were created by the chemical modification of the test substrate polymers; nylon and ethylvinyl alcohol (EVOH). Surfaces were chemically modified by the covalent coupling of antimicrobial agents known to be active against the ...

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Analysis of the pattern of the chromosomal localization of quantitative trait loci (QTLs) is necessary for comprehensively understanding their functions. The chromosomal localization of QTLs controlling milk production traits has been studied in cattle chromosomes. The distribution of QTLs between chromosomes has proved to be binomial. Their distribution along each chromosome was, in general, uniform, except for the QTLs controlling the somatic cell score (SCS), which tended towards telomeric location. However, there are chromosomes either enriched with or particularly poor in QTLs. The QTL distribution patters are the most similar for the milk yield (M) and milk protein yield (P) and for milk fat yield (F) and milk fat content (%F). The pattern of the SCS QTLs stands out among those of ot...

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In response to a heat shock, the yeast Saccharomyces cerevisiae undergoes a large increase in its resistance to heat and, by the induction of its recombinational DNA repair capacity, a corresponding increase in resistance to radiation. Yeast which lack mitochondrial DNA, mitochondria-controlled protein synthetic apparatus, aerobic respiration, and electron transport (rho/sup 0/ strain) were used to assess the role of O/sub 2/, mitochondria, and oxidative processes controlled by mitochondria in the induction of these resistances. We have found that rho/sup 0/ yeast grown and heat shocked in either the presence or absence of O/sub 2/ are capable of developing both radiation and heat resistance. We conclude that neither the stress signal nor its cellular consequences of induced heat and radiation resistance are directly dependent on O/sub 2/, mitochondrial DNA, or mitochondria-controlled protein synthetic or oxidative ...

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Replication-defective vectors derived from reticuloendotheliosis virus were used to transduce exogenous genes into early somatic stem cells of the chicken embryo. One of these vectors transduced and expressed the chicken growth hormone coding sequence. The helper cell line, C3, was used to generate stocks of vector containing about 10/sup 4/ transducing units per ml. Injection of 5- to 20-..mu..l volumes of vector directly beneath the blastoderm of unincubated chicken embryos led to infection of somatic stem cells. Infected embryos and adults contained unrearranged integrated proviral DNAs. Embryos expressed the transduced chicken growth hormone gene and contained high levels of serum growth hormone. Blood, brain, muscle, testis, and semen contained from individuals injected as embryos contained vector DNA. Replication-defective vectors of the reticuloendotheliosis virus transduced exogenous genes into chicken embryonic stem cells in vivo.

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Pectin rich cold stored spoiled fruits, vegetables and cold soils were screened and different pectinolytic isolates were obtained by enrichment culturing and ruthenium red plate assay. Among the primary isolates 10-15% were yeast isolates. Six isolates with higher zones of pectin hydrolysis were selected and tested for polygalacturonase (PGU) production at room temperature (25 degrees C) and at 5 degrees C. One isolate identified as Saccharomyces sp. with highest polygalacturonase activity at 5 degrees C was used for enzyme production using raw fruit pectins as substrates. The isolate was identified by preliminary cultural, morphological and sugar fermentation tests. PGU production was high in raw pectin substrates like orange peel (21 U/ml), apple peel (20 U/ml ), mango peel (19 U/ml), ...

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A 65-yr-old female developed cough, fever and dyspnoea following repeated exposure to a home ultrasonic humidifier. High-resolution computed tomography showed ground-glass opacity in both lung fields. Arterial blood gas analysis gave an oxygen tension of 8.38 kPa (63 Torr). Pulmonary function testing revealed restrictive ventilatory impairment with a reduction in the diffusing capacity. The diagnosis of extrinsic allergic alveolitis (EAA) was confirmed by radiographic findings, pathological evidence of alveolitis and reproductive development by a provocation test to the humidifier water. The yeast Debaryomyces Hansenii was the only microorganism cultured from the water of the humidifier. The double diffusion precipitating test and lymphocyte proliferative response was positive for an extract of D. Hansenii, providing evidence to incriminate this fungus. This is the first described case of EAA caused by D. Hansenii. PMID:12449192

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Experiments were designed to elucidate whether or not the irradiated bone marrow cells receive any stimulation for the self-replication and differentiation from normal 'stromal' cell colonies in the bone marrow cell culture in vitro. When irradiated or unirradiated bone marrow cells were overlaid on the normal adherent cell colonies, the proliferation of haemopoietic stem cells was supported, the degree of the stimulation depending on the starting cellular concentration. There was, however, no significant changes in the concentration of either CFUs or CFUc regardless of the dose of irradiation on the bone marrow cells overlaid. This was a great contrast to the dose-dependent decrease of CFUs or CFUc within the culture in which both the stem cells and stromal cells were simultaneously irradiated. These results suggest that the balance of self-replication and differentiation of the haemopoietic stem cells is affected only when ...

UK PubMed Central (United Kingdom)

Ribosomal protein L1 from Saccharomyces cerevisiae binds 5S rRNA and can be released from intact 60S ribosomal subunits as an L1-5S ribonucleoprotein (RNP) particle. To understand the nature of the...Full Text Available

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In order to study the intragenic profiles of active transcription, we determined the relative levels of active RNA polymerase II present at the 3′- and 5′-ends of 261 yeast genes by...Full Text Available

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A refined map for the linear arrangement of histones along DNA in nucleosomal core particles has been determined by DNA-protein crosslinking. On one strand of 145-bp core DNA, histones are aligned in...Full Text Available

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Chapter 5, describes some of the most important molecular methods used in the study of chromosome structure and function. The methods discussed include fragmentation of DNA, cloning, flow cytometry and chromosome sorting, is situ hybridization, polymerase chain reaction (PCR), and yeast artificial chromosomes (YACs). 18 refs., 3 figs., 1 tab.

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Procedures are described for the use of synthetic oligonucleotides for Southern blot experiments and gene bank screening, and the effect of various mismatches on the efficiency of hybridization is demonstrated. The following topics are discussed: sensitivity vs. specificity, hybridization of a 12-mer to the lambda endolysin gene; hybridization of oligonucleotide probes to the E. coli lac operator; hybridization of synthetic probes to the CYC1 gene of yeast; and cloning eucaryotic genes. (HLW)

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Acyl-CoA:diacylglycerol acyltransferase (EC 2.3.1.20) is a membrane protein present mainly in the endoplasmic reticulum. It catalyzes the final and committed step in the biosynthesis of triacylglycerol,...Full Text Available

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Current investigations into the biological solubilization of coal with microorganisms focus on the production of solubilizing activity in fungi. Test organisms for this work include a species of the yeast Candida previously isolated from a lignite outcrop, and P. chrysosporium, a filamentous higher fungus which has played a major role in lignin biodegradation research. The studies described are primarily exploratory in nature, and are fundamental to the design of more sophisticated inquiries into the physiology of fungal coal solubilization.

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BackgroundHIV infection causes a qualitative and quantitative loss of CD4⁺ T cell immunity. The institution of anti-retroviral therapy (ART) restores CD4⁺ T...Full Text Available

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In late mitosis and G1, Mcm2-7 are assembled onto replication origins to license them for initiation in the upcoming S phase. After initiation, Mcm2-7 provide helicase activity to unwind DNA at the...Full Text Available

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Differential host-pathogen interactions direct viral replication in infected cells. In HIV-1 infected cells, nuclear export of viral RNA transcripts into cellular cytoplasm is governed by interaction...Full Text Available

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Following infection with the protozoan parasite Leishmania major, C57BL/6 mice develop a small lesion that heals spontaneously. Resistance to infection is associated with the development...Full Text Available

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Ethidium bromide (EB) and ultraviolet light (UV) in combination are known to produce a synergistic induction of 'petite' mutants in yeast. Two other agents were combined with EB, 3-Carbethoxypsoralene (3 CPs) activated by 365 nm light or #gamma# rays. EB in combination with 3 CPs also resulted in an enhanced production of 'petite' mutants. After the photoaddition of 3 CPs in exponential phase cells, recovery of the 'petite' mutation during dark liquid holding was inhibited by the presence of EB producing an enhanced number of 'petite' mutants. The behavior of mitochondrial antibiotic resistance markers after individual and combined treatments with EB and 3 CPs indicates a random loss of markers after EB and a preferential loss of a certain region for the 3 CPs photoaddition. The combination of the two agents leads to an additivity of total drug marker losses rather than a synergistic loss. The combination of EB with #gamma# rays produced no enhancement in 'petite' ...

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Papers in this book illustrate the utility of mineral biotechnology with respect to biobeneficiation, bioleaching, bioremediation and biomineralization. Papers of particular interest to the coal industry include: depression of pyrite flotation by yeast and bacteris (S.K. Kawatra and T.C. Eisele); desulfurization of coal by microbial flotation in a semicontinuous system (T. Nagaoka and others); biochemical removal of HAP precursors from coal - INEEL slurry column testing (K.S. Noah and G.J. Olson); microorganisms, biotechnology and acid rock drainage - emphasis on passive-biological control and treatment methods (N. Kuyucak); and utility of bioreagents in mineral processing (P. Somasundaran and others).

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The biodegradation of ring-"1"4C- and methyl-"1"4C-labeled paraquat by the soil yeast Lipomyces starkeyi was studied in vitro. It was found that the degradation of paraquat (acting as a sole source of culture nitrogen) resulted in the accumulation in the extracellular medium of radiolabeled acetic acid. The culture also evolved radiolabeled CO_2. The results suggest that the degradation of paraquat by L. starkeyi is associated with the integrity of the cell wall and that disruption or removal of the wall results in a complete loss of degradative capability. A mechanism for the degradation of paraquat by this organism is postulated.

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Glabridin, an active constituent of Glycyrrhiza glabra roots, was found to be active against both yeast and filamentous fungi. Glabridin also showed resistance modifying activity against drug resistant mutants of Candida albicans at a minimum inhibitory concentration of 31.25-250 g/mL. Although the compound was reported earlier to be active against Candida albicans, but this is the first report of its activity against drug resistant mutants. Copyright Copyright 2009 John Wiley & Sons, Ltd.

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Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck (Piperaceae) is a medicinal plant traditionally used in Brazil to treat infectious diseases. The extracts obtained from the leaves of P. regnellii were investigated for their antifungal activities against the yeasts Candida albicans, C. krusei, C. parapsilosis, and C. tropicalis. The EtOAc extract presented gnificant activity against Candida albicans with MIC at 125 {mu}g mL{sup -1}, and a moderate activity against both C. krusei and C. parapsilosis with MIC at 500 {mu}g mL{sup -1}. Candida tropicalis was not inhibited by this extract at concentrations as high as 1000 {mu}g mL{sup -1}. Based on these findings, the EtOAc extract was fractionated by silica gel column chromatography into nine fractions. The hexane and CHCl{sub 3} fractions showed varied levels of antifungal activity against all test yeasts. Further column chromatography separation of the hexane fraction afforded the pure ...

International Nuclear Information System (INIS)

The paper is concerned with the simulation of practical testing situations which are too difficult and/or expensive to replicate in a laboratory environment. Numerical experiments are described which simulate the differential eddy current probe response to the build-up and chemical flushing of magnetite in the crevice gap of a PWR steam generator unit. The simulation results agree well with the only experimental data available to the authors and lead to the conclusion that conventional differential eddy current probes should be capable of characterizing crevice gap conditions with respect to the presence of magnetite. (author).

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Purpose: The purpose of this study was to evaluate the efficacy and toxicity of replication-competent adenovirus-mediated double suicide gene therapy in an adjuvant setting with external beam radiation therapy (EBRT) in an experimental prostate cancer model in preparation for a Phase I clinical study in humans. Methods: For efficacy studies, i.m. DU145 and intraprostatic LNCaP C4-2 tumors were established in immune-deficient mice. Tumors were injected with the lytic, replication-competent Ad5-CD/TKrep adenovirus containing a cytosine deaminase (CD)/herpes simplex virus thymidine kinase (HSV-1 TK) fusion gene. Two days later, mice were administered 1 week of 5-fluorocytosine + ganciclovir (GCV) prodrug therapy and fractionated doses of EBRT (trimodal therapy). Tumor control rate of trimodal therapy was compared to that of EBRT alone. For toxicology studies, immune-competent male mice received a single intraprostatic injection (10"1"0 vp) of the ...

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Glioblastoma multiforme is the most aggressive form of human brain tumor, which has no effective cure. Previously, we have demonstrated that overexpression of the C-terminal fragment of the human telomerase reverse transcriptase (hTERTC27) inhibits the growth and tumorigenicity of human cervical cancer HeLa cells. In this study, the therapeutic effect and molecular mechanisms of hTERTC27-mediated cancer gene therapy were further explored in vivo in established human glioblastoma xenografts in nude mice. We showed that intratumoral injection of adeno-associated virus carrying hTERTC27 (rAAV-hTERTC27) is highly effective in reducing the growth of the subcutaneously transplanted glioblastoma tumors. Histological analyses showed that rAAV-hTERTC27 treatment leads to profound necrosis, apoptosis, infiltration of polymorphonuclear neutrophils and reduced microvessel density in the tumor samples. To study the molecular mechanism of rAAV-hTERTC27-mediated antitumor effects, we analyzed the ...

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Wound dressings impregnated with silver have a role to play in aiding to reduce both the dressing and wound microbial bioburden. It is therefore imperative that antimicrobial wound dressings have efficacy on a broad range of clinical significant microorganisms. Accordingly, this study aimed to determine the antimicrobial efficacy of a silver alginate dressing against 115 wound isolates that had been isolated routinely from patients at West Virginia University Hospital. Standardised corrected zones of inhibition (CZOIs) were performed on all clinical isolates. It was found that the silver alginate dressing was able to inhibit the growth of all microorganisms tested. In particular, the silver alginate dressing inhibited the growth of Candida albicans and yeasts with CZOI of 3-115 mm. All met...

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Mono- and disubstituted novel derivatives of the heptaene nystatin analog 28,29-didehydronystatin A1 (S44HP, 1) were obtained by chemical modification of the exocyclic C-16 carboxyl and/or an amino group of mycosamine moiety. The strategy of preparation of mono- and double-modified polyene macrolides was based on the use of intermediate hydrophobic N-Fmoc (9-fluorenylmethoxycarbonyl) derivatives that facilitated the procedures of isolation and purification of new compounds. The antifungal activity of the new derivatives was first tested in vitro against yeasts and filamentous fungi, allowing the selection of the most active compounds that were subsequently tested for acute toxicity in mice. 2-(N,N-dimethylamino)ethylamide of 1 (2) and 2-(N,N-dimethylamino)ethylamide of N-fructopyranosyl-28...

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Background: The environment preferred by Acanthamoeba trophozoites and the mechanism by which the amebae enters the cornea are not yet fully understood. A better understanding of the pathogenesis of this disease may help with prevention and treatment. Purpose: To define the preferred environments for Acanthamoeba survival and proliferation in vitro by examining the effect of tonicity, nutrient concentration, and free chlorine content on Acanthamoeba. Materials and methods: Human corneal isolates of Acanthamoeba castellanii and Acanthamoeba polyphaga trophozoites were cultured at 22^oC (room temperature) in PYG (peptone-yeast extract-glucose) medium. The effect of tonicity on amebae was determined by incubating trophozoites in sodium chloride solutions in concentrations ranging from 0% to 1...

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Statistically based experimental designs were applied to screen and optimize the bioleaching of spent hydrocracking catalyst by Penicillium simplicissimum. Eleven factors were examined for their significance on bioleaching using a Plackett-Burman factorial design. Four significant variables (pulp density, sucrose, NaNO"3, and yeast extract concentrations) were selected for the optimization studies. The combined effect of these variables on metal bioleaching was studied using a central composite design (CCD). Second-order polynomials were established to identify the relationship between the recovery percent of the metals and the four significant variables. The optimal values of the variables for maximum metals bioleaching were as follows: pulp density (4.0%, w/v), sucrose (90g/L), NaNO"3 (2...

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Direct plating, selective enrichment, and cold enrichment followed by secondary selective enrichment procedures were compared for detecting and enumerating Listeria monocytogenes in chopped cabbage stored at 5 degrees C for up to 64 days. Addition of Fe3+ to solid media enhanced detection of the organism. Cold enrichment (5 degrees C) in nutrient broth and brain heart infusion broth followed by secondary enrichment (48 h, 30 degrees C) in Trypticase soy-yeast extract-antibiotic broth and thiocyanate-nalidixic acid broth and plating on selective agar media (Doyle and Schoeni selective enrichment agar [minus acriflavin hydrochloride, supplemented with 5 micrograms of Fe3+/ml] and McBride Listeria agar) resulted in the detection of highest populations. PMID:3111369

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The potential of the yeast Candida tropicalis and Candida guillermondii was evaluated and an isolated partnership of microorganisms of waters of the Medellin River, conformed by two bacteria and one leavening, to degrade the content of organic matter present in wine produced by the factory of Licores and Alcoholes de Antioquia (FLA) in aerobic process with biomass production. For each one of the microorganisms in study this capacity of removal in units of chemical demand of oxygen was quantified (CDO); in addition, parameters were analyzed such as yield of the biomass in relation to the removed CDO and to total reducing sugars (TRS) consumed, time of fermentation and speed of growth different dilutions from wine. Also the possible inhibition was analyzed that the present phenolic compounds in this wine can cause in the biological process of degradation.

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In this work, silver-montmorillonite (Ag-MMT) antimicrobial nanoparticles have been obtained by allowing silver ions from nitrate solutions to replace the Na^+ of natural montmorillonite and then to be reduced by a thermal treatment. Ag-MMT were used as active antimicrobial compounds to improve the shelf life of fresh fruit salad. In order to assess their influence on product shelf life, sensorial and microbiological quality has been monitored during the storage. The microbiological quality was determined by monitoring the principal spoilage microorganisms (mesophilic and psychrotrophic bacteria, coliforms, lactic acid bacteria, yeasts and molds). Additionally, the evolution of sensorial quality was assessed by monitoring color, odor, firmness and product overall quality. The Ag-MMT nanopa...

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A number of aerobic species capable of decolorizing some of the dyes in a textile mill effluent were isolated. One of the isolates was able to decolorize Terasil black dye under aerobic conditions in the presence of an exogenous carbon source after 5 days. Glucose or starch (%1 ea) are essential for decolorization but the process proceeds faster in the presence of 0.5% yeast extract. Results of the BOD5 show that the untreated effluent samples have a low BOD value, whereas treated samples show an initial increase in BOD up to 15 days followed by a decrease after 20 days. FT-IR and GC-MS data also reveal that the initial components in the untreated effluent disappear after 20 days of treatment, confirming biodegradation of the dye. Phytotoxicity tests on the untreated effluent samples using...

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This review describes how a mismatch between the knowledge produced by scientists and the evidence demanded by regulators has emerged, and how society has struggled to find definitive answers to questions of safety, for an important flame retardant chemical in current use ? Deca-BDE. This has involved two key disciplines: analytical chemistry and toxicology. Within the chemistry, a lack of standardized methodologies among scientists has resulted in a persistent yet largely undeclared failure to replicate results within the discipline. Within the toxicology, the quest for innovative, curiosity-driven research by university scientists in preference to using validated standard protocols, designed to promote consistency within the risk assessment process, has prompted questions about the credi...

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Sleep symptoms are a prominent feature of mental health disorders like PTSD and depression. However, it is unknown whether sleep symptoms mediate the relationship between combat stress and these disorders. We examined the mediating role of sleep symptoms on the relationship between combat stress and PTSD; and the relationship between combat stress and depression using data from 576 Army veterans of the Iraq War surveyed in 2004. Correlational analyses revealed that when insomnia was included in the model, the correlation between combat stressors and other depression symptoms decreased by 65%; and when nightmares were included in the model, the correlation between combat stressors and other PTSD symptoms decreased by 69%. We replicated these analyses using individual items assessing PTSD an...

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This research proposes that the concept of emotional attachment, and specifically the independent constructs of psychological ownership and affective reaction, can help explain many of the endowment effect findings documented in the literature. We define these constructs and then test them across a set of nine studies in which we both replicate previous and generate new endowment effect findings, and then show that psychological ownership and affective reaction can mediate the effects. In doing so, we offer direct empirical support for the idea of emotional attachment as a driver of loss aversion while also providing practitioners and future endowment effect researchers with new insights about the psychological processes that underlie the endowment effect.

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Highly ordered arrays of Ni nanoholes and Fe{sub 20}Ni{sub 80} antidots have been prepared, respectively, by replica/antireplica processing and sputtering techniques using nanoporous alumina membranes as templates. Geometrical characteristics as nanohole/antidot diameter, interpore distance and the overall hexagonal symmetry of arrays are controlled through the original templates. Experimental data on their hysteresis and magnetic domain structure have been taken by vibrating sample magnetometry and magnetic force microscopy, respectively. An analysis of the magnetization process, resulting magnetic anisotropy and magnetic domain structure is summarized considering the influence of those geometry aspects. In particular, the hexagonal symmetry and the density of nanohole/antidots determine the overall magnetic behavior, which is of interest in future high-density magnetic storage systems.

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Seed priming technique has been known to improve germination and seedling emergence under different environment stresses. The objective of this study was to evaluate the effectiveness of seed priming in improving the seed germination and seedling vigor of Agropyron elongatum and Bromus inermis in response to five concentrations of the allelopathic extract of Thymus kotschyanus (0, 5%, 25%, 50% and 75%) under laboratory conditions. Pretreatments included gibberellic acid (250, 500 and 1000 ppm), potassium nitrate (0.1%, 0.2% and 0.3%) and salicylic acid (100, 200 and 300 mg/L). This experiment was carried out as a factorial experiment based on a completely randomized design with four replications. The results showed that seed germination, germination speed and seedling vigor were significan...

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We investigated the impacts of forest thinning, prescribed fire, and contour ripping on community level physiological profiles (CLPP) of the soil microbial population in postmining forest rehabilitation. We hypothesized that these management practices would affect CLPP via an influence on the quality and quantity of soil organic matter. The study site was an area of Jarrah (Eucalyptus marginata Donn ex Sm.) forest rehabilitation that had been mined for bauxite 12?years previously. Three replicate plots (20???20?m) were established in nontreated forest and in forest thinned from 3,000?8,000 stems ha?1 to 600?800 stems ha?1 in April (autumn) of 2003, followed either by a prescribed fire in September (spring) of 2003 or left nonburned. Soil samples were collected in August 2004 from two soil ...

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Aims We tested cardiovascular and visceromotor reflex (VMR) responses to urinary bladder distension (UBD) in urethane anesthetized rats to see if it can replicate the response pattern and the inhibition of bladder nociceptive transmission by analgesics seen in isoflurane anesthetized animals. Methods Female Sprague-Dawley rats under 3% isoflurane anesthesia were acutely instrumented with jugular venous, carotid arterial, and bladder cannulas for drug administration, blood pressure (BP) measurement, and bladder distension, respectively. Needle electrodes were placed directly into the abdominal musculature to measure myoelectrical activity subsequent to phasic UBD (30 sec in 3 min intervals). A cardiovascular response (pressor) and a VMR response (a contraction of abdominal and hind limb mus...

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Nitrogen use efficiency (NUE) of tef, a major staple crop in Ethiopia, is very low, either caused by untimely use of nitrogen (N) fertilizers or lack of other essential nutrients like sulphur (S). The average grain yield of this crop is low, averaging ?1 in farmer?s fields of the semi-arid conditions. Therefore, the present study was conducted to see the effect of the timing of combined N and S fertilization on the yield, yield components, and N and S concentration in the plant parts of the crop. A factorial combination of three rates of N (0, 70, and 105?kg?ha?1) with four rates of S (0, 16, 32, and 48?kg?ha?1) was applied in randomized complete blocks in three replications. The experiment was carried out in the 2004 and 2005 cropping seasons in the Cambisols of the semi-arid area of Ethi...

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The objectives of the project were to make significant improvements in the production of stall regulated blades in the areas of (a) the tip box, its housing, its mechanism and small GRP parts; (b) mould technology; (c) resins and glues and (d) root tooling. Although wood composite had been identified as a competitive technology for blades, compared with GRP blades, production volumes had been lower; reasons are given. The way in which the four areas identified for investigation were tackled are discussed. The study showed that the mould cycle time can be reduced to two days for a stall regulated blade and the blade quality can be improved by using the composite tip box and new resins. The time required for replication of moulds can be reduced by 40%.

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Banana bunchy top virus (BBTV), family Nanaviridae, genus Babuvirus, is a single stranded DNA virus (ssDNA) that causes banana bunchy top disease (BBTD) in banana plants. It is the most common and most destructive of all viruses in these plants and is widespread throughout the Asia-Pacific region. In this study we isolated, cloned and sequenced a BBTV sample from Hainan Island, China. The results from sequencing and bioinformatics analysis indicate this isolate represents a satellite DNA component with 12 DNA sequences motifs. We also predicted the physical and chemical properties, structure, signal peptide, phosphorylation, secondary structure, tertiary structure and functional domains of its encoding protein, and compare them with the corresponding quantities in the replication initiatio...

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Abstract Recent reports showed many patients with chronic fatigue syndrome (CFS) harbor a retrovirus, xenotropic murine leukemia-related virus (XMRV), in blood; other studies could not replicate this finding. A useful next step would be to examine cerebrospinal fluid, because in some patients CFS is thought to be a brain disorder. Finding a microbe in the central nervous system would have greater significance than in blood because of the integrity of the blood-brain barrier. We examined cerebrospinal fluid from 43 CFS patients using polymerase chain reaction techniques, but did not find XMRV or multiple other common viruses, suggesting that exploration of other causes or pathogenetic mechanisms is warranted. Ann Neurol 2011;

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Ethanol has excellent fuel properties, such as high octane, high heat of vaporization and low photochemical reactivity in the atmosphere. It is less volatile than gasoline and there is lower smog formation from evaporative emissions of pure ethanol compared to gasoline. As such, ethanol has emerged as an important alternative energy source that is sustainable, efficient, cost effective, convenient and safe. In 2006, global production of ethanol reached 13.5 billion gallons, up from 12.1 billion gallons in 2005. However, in light of the current debate of food versus fuel, the industry must shift to non-food feedstocks. This paper described an emerging technology to cost-effectively produce ethanol from sweet sorghum stalks, the most promising alternative feedstock to corn, via solid state fermentation (SSF). Experiments of advanced solid state fermentation (ASSF) for ethanol production from sweet sorghum by Saccharomyces cerevisiae were conducted in laboratory and pilot scales studies. ...

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Reclamation of copper mine tailings in a cost effective, successful, and sustainable manner is an ongoing area of evaluation in the arid southwest. A study was initiated in September, 1996 near Hayden, Arizona to evaluate the use of municipal biosolids for reclaiming acidic copper mine tailings (pH of 2.5 to 4.0). The main objectives of the study were to (1) define an appropriate level of biosolids application for optimum plant growth, and (2) evaluate the effects of green waste and lime amendments. The experiment was a randomized complete block design with four biosolid rates of 20, 70, 100 and 135 dry tons/acre, three amendment treatments (none, green waste, and green waste plus lime); with three replications. Non-replicated controls (no treatment, green waste only and lime only) were included for comparison. Shortly after biosolids incorporation to a depth of 10--12 inches, composite soil samples (0--12 inches) of each plot were taken. ...

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The ability of residual biomass from the thermotolerant ethanol-producing yeast strain Kluyveromyces marxianus IMB3 to function as a biosorbent for uranium has been examined. It was found that the biomass had an observed maximum biosorption capacity of 120 mg U/g dry weight of biomass. The calculated value for the biosorption maximum, obtained by fitting the data to the Langmuir model was found to be 130 mg U/g dry weight biomass. Maximum biosorption capacities were examined at a number of temperatures and both the observed and calculated values obtained for those capacities increased with increasing temperature. Decreasing the pH of the biosorbate solution resulted in a decrease in uptake capacity. When biosorption reactions were carried out using sea-water as the diluent it was found that the maximum biosorption capacity of the biomass increased significantly. Using transmission electron microscopy, uranium crystals were shown to be concentrated on the outer ...

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Transcriptional regulation of the galactose-metabolizing genes in Saccharomyces cerevisiae depends on three core proteins: Gal4p, the transcriptional activator that binds to upstream activating DNA sequences (UASGAL); Gal80p, a repressor that binds to the carboxyl terminus of Gal4p and inhibits transcription; and Gal3p, a cytoplasmic transducer that, upon binding galactose and adenosine 5'-triphosphate, relieves Gal80p repression. The current model of induction relies on Gal3p sequestering Gal80p in the cytoplasm. However, the rapid induction of this system implies that there is a missing factor. Our structure of Gal80p in complex with a peptide from the carboxyl-terminal activation domain of Gal4p reveals the existence of a dinucleotide that mediates the interaction between the two. Biochemical and in vivo experiments suggests that nicotinamide adenine dinucleotide phosphate (NADP) plays a key role in the initial induction event.

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All organisms, from bacteria and yeast to humans, respond to physical and chemical stressors by increasing the synthesis of a small group of cellular stress proteins.'' The authors have developed a simple in vitro system for quickly screening environmentally relevant stressors to detect stress-induced proteins that are good candidates for biomarkers. Polyacrylamide gel electrophoresis was used to detect stressor-induced, concentration-dependent changes in cellular stress protein levels in two fish cell culture systems, whereas simultaneous in vitro neutral red uptake cytotoxicity assays measured the stressors effect on cellular physiology. There was a direct concentration-dependent relationship between sublethal cytotoxic effects and the increases in stress protein levels. Increases of 50 to 200% were detected in stress proteins from desert topminnow, Poeciliopsis lucida, hepatoma-derived cell cultures exposed to cadmium or copper. Three proteins ...

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The authors describe a cloning strategy for the construction of a human genomic library in yeast artificial chromosomes (YACs) based on complete digestion of high-molecular-weight DNA with the infrequently cutting restriction enzyme MluI. Cloning of MluI fragments in the vector pYAC-RC and one subsequent size fractionation by preparative pulsed-field gel electrophoresis yielded a library with average insert sizes of 600 kb. Ninety-seven percent of the colonies were recombinant. An additional size fractionation of MluI fragments prior to ligation had no significant influence on the size of the YACs. The library currently consists of 5000 clones, which is the equivalent of one human genome. Nineteen percent of the YACs were larger than 1.2 Mb. Since smaller MluI fragments are lost during sizing, they also performed cloning without size fractionation. Only 20% of the colonies were recombinant, probably due to unligated vector fragments that were present during the ...

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The potential of the dried yeast, wild-type Schizosaccharomyces pombe, to remove Ni(II) ion was investigated in batch mode under varying experimental conditions including pH, temperature, initial metal ion concentration and biosorbent dose. Optimum pH for biosorption was determined as 5.0. The highest equilibrium uptake of Ni(II) on S. pombe, q e, was obtained at 25??C as 33.8?mg?g?1. It decreased with increasing temperature within a range of 25?50??C denoting an exothermic behaviour. Increasing initial Ni(II) concentration up to 400?mg?L?1 also elevated equilibrium uptake. No more adsorption took place beyond 400?mg?L?1. Equilibrium data fitted better to Langmuir model rather than Freundlich model. Sips, Redlich?Peterson, and Kahn isotherm equations modelled the investigated system with a...

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The large girth and immense size of the baobab has caused many to speculate about its age. Unfortunately reliable age estimates cannot be determined from growth rates as the girth varies in response to different moisture regimes. In a similar way, ages cannot be determined from ring-width measurements or X-ray densitometry as the absorbent nature of the soft fibrous wood and distortion upon drying prevent the application of these techniques. The Southern Hemisphere bomb radiocarbon curve was used to demonstrate that the rings of a recently-fallen baobab (Adansonia digitata L.) from Kruger National Park appear to be annual. The detrended C isotope values of finely-ground wholewood from another baobab specimen were found to be highly associated with January precipitation (r = 0.72; p < 0.01). This study demonstrates that high resolution information about past climates may be obtained by analysing the C isotope values from baobab samples even if distortion of ring-widths has ...

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In this study, the complete sequence of the mitochondrial DNA (mtDNA) of Atelura formicaria (Hexapoda: Zygentoma) is described. The molecule is 15.205 bp in length and it is the third complete mt genome sequenced from the Zygentoma. The genome organization conforms with the putative ancestral insect gene arrangement. All protein coding genes use standard initiation codons (methionine and isoleucine). The exception is nad4 that starts with GTG, a codon used for this purpose in other insect species. A peculiar strand skew bias is observed, given that the PCGs encoded on the J-strand contain more thymines than adenines and more cytosines than guanines. This trend in nucleotide composition has been observed also in the "firebrat" Thermobia domestica (Zygentoma, Lepismatidae), but differs from that of the majority of hexapod species, including Tricholepidion gertschi (Zygentoma, Lepidotrichidae), where adenines and cytosines outnumber thymines and guanines. The presence of structural ...

International Nuclear Information System (INIS)

The economic attractiveness and negative environmental impact of nitrogen (N) fertilization in pastures depend on the N use efficiency in the soil-plant system. However, the recovery of urea-"1"5N by Panicum maximum cv. Tanzania pastures, one of the most widely used forage species in intensified pastoral systems, is still unknown. This experiment was conducted in a randomized complete block design with four treatments (0, 40, 80 and 120 kg ha-1 of N-urea) and three replications, to determine the recovery of "1"5N urea by Tanzania grass. Forage production, total N content and N yield were not affected by fertilization (p > 0.05), reflecting the high losses of applied N under the experimental conditions. The recovery of "1"5N urea (% of applied N) in forage and roots was not affected by fertilization levels (p > 0.05), but decreased exponentially in the soil and soil-plant system (p < 0.05) with increasing urea doses. The amount of "1"5N (kg ha"-"1) in ...

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This report briefly describes the studies on the mechanism of in vivo DNA repairing by the author in Research Reactor Institute, Kyoto Univ. for the past 30 years. First, the ability of UV radiation to induce transformation was investigated with viral DNA. The formation of thymine-thymine dimer was found harmful to organisms and such dimers were removable by UV-radiation at a low frequency. The mutability was determined in three different E.coli strains with mutator gene, mutT, mutS or mutL. The ability to excise 8-oxoguanin developed in primer DNA was deficient in mutT and miss-pairing left after DNA replication could not be recovered in mutL and mutS strains. Further, DNA repairing mechanism was investigated in other microorganisms; single-strand cleavage caused by exposure to BNCB radiation (boron-neutron-captured beam) could not be repaired in E. coli. Whereas for Deinococcus radiodurans, of which survival rate was not decreased by #gamma#-ray radiation at 5 ...

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We investigated the Raman coherence characteristics of a solid hydrogen film deposited on a sapphire substrate held at 5.3 K. Using Raman coherence prepared with two single-frequency pulsed lasers, we generated the multiorder coherent Raman sidebands in solid hydrogen film. The highorder Raman sidebands were obtained under strong pumping intensities (? 230 MW/cm2). The generated anti-Stokes(AS)-Raman sidebands extend from the ultraviolet (292 nm for the AS5 band) to the visible (565 nm for the AS1 band) region. The multiorder Raman sideband generation is found to be due to the parametric coupling of pump and coupling lasers. The frequency conversion efficiency from pumping beams to the first AS-Raman sideband shows a maximum (14 %) at a pumping intensity of 360 MW/cm2. From an experiment that makes the multimode probe beam beat with the prepared Raman coherence, we found that the prepared Raman coherence replicates the probe beam to its Raman sidebands.

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Endoreduplication represents a cell cycle variant during which multiple rounds of DNA replication occur without subsequent chromosome separation and cytokinesis, resulting into a cellular increase of the DNA content. Although the DNA ploidy level of cells is controlled by external stimuli such as light, currently limited knowledge is available on how environmental signals regulate the endoreduplication cycle at the molecular level. Previously, we have demonstrated that the conversion from the mitotic cell cycle into an endoreduplication cycle is mediated by the atypical E2F transcription factor DEL1 that operates as a repressor of endocycle onset. Here, we identified DEL1 as a transcriptional target of the classical E2Fb and E2Fc transcription factors that antagonistically control DEL1 transcript levels through competition for a single E2F cis-acting binding site. Correspondingly with the reported opposite effect of light on the E2Fb and E2Fc protein levels, the ...

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Abstract Sam68 (Src-associated protein in mitosis 68-kDa) is a multifunctional protein, known to govern cellular signal transduction, transcription, RNA metabolism, proliferation, apoptosis, and HIV-1 replication. Although intrinsic mechanisms that modulate Sam68 function are beginning to emerge, the regulatory events contributing to its expression remain elusive. We previously reported that heat shock protein-22 (Hsp22) antagonizes Sam68 function in rev-response element (RRE)-mediated gene expression. We now demonstrate that Sam68 levels correlate inversely with Hsp22 in a variety of cells, including U87, Jurkat, 293T, and U-937. In U87 glioblastoma cells, which contained high levels of Hsp22 than other cell lines tested, Hsp22 knockdown dramatically increased both Sam68 mRNA and protein,...

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The SCILS program investigates and demonstrates concepts of operation for dispersed theater forces C3. The dispersal concept would increase the number of potential targets for nuclear strike, make these targets smaller, more highly mobile and harder to find, and introduce functional redundancy. The SCILS system is a microcomputer-based approach to preserving and enhancing the effectiveness of C3 operation in a dispersed configuration. In this program, the system was implemented for testing for the 9th Infantry Division in coordination with the Army's Distributed Command and Control System. Technical highlights of the system include a distributed, replicated data base, a video map/graphics-oriented Commander's Decision Display System, a packet switched Multi-Media Network that operates with deployed field communication systems, and a Voice/Data Conferencing System that supports remote coordination/briefing in a dispersed configuration. The program ...

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Acetylcholine and other neurotransmitters play unique trophic roles in brain development. Accordingly, drugs and environmental toxicants that promote or interfere with neurotransmitter function evoke neurodevelopmental abnormalities by disrupting the timing or intensity of neurotrophic actions. The current review discusses three exposure scenarios involving acetylcholine systems: nicotine from maternal smoking during pregnancy, exposure to environmental tobacco smoke (ETS), and exposure to the organophosphate insecticide, chlorpyrifos (CPF). All three have long-term, adverse effects on specific processes involved in brain cell replication and differentiation, synaptic development and function, and ultimately behavioral performance. Many of these effects can be traced to the sequence of cellular events surrounding the trophic role of acetylcholine acting on its specific cellular receptors and associated signaling cascades. However, for chlorpyrifos, additional ...

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The present work's objective is to test sampling methodologies applied on the bioremediation processes in situ, involving the establishment of sample replicates, specific laboratory procedures and its results, inedited on the referred technique. Agricultural fertilizers were used, NPK and OSMOCOTE, as biostimulants on mangroves substrates affected by petroleum activities. The tested methodology used on the experiment was based on the monitoring of this technology in aquarium with water from Sao Paulo's river and sediment contaminated by oil, realized in three steps (first pre-test, second pre-test and third pre-test) that happened between the months of August and November of 2007. The physical-chemical parameters were measured with portable devices carefully calibrated and the oil analyzed with gas chromatography. The saturated hydrocarbons (n-alkenes) had an increase on the concentrations for some aquariums and these found results do not permit ...

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In the third millennium, when the main words are globalization, world net of computers, genetic code and efficiency, deep transformations have been incorporated to human behavior. Among such transformations it may be pointed out, matching with the objectives of this study, the modem consumer's profile. The consumers have quickly become more and more conscious and demanding regarding to the quality of the available products in the market. In that sense, the consumer's concern about the natural and healthy appearance, as well as the preservation of the nutritious content of the foods, have been a constant sign of alert for producers and suppliers. Besides, the timeless for domestic preparation of foods have imposed the use of minimally processed as an incontestable reality. A barrier for the production of minimally processed foods is imposed by the degree of perishability of certain products, such as the cassava-parsley. This vegetable is a source of energy, calcium, phosphorus and ...

International Nuclear Information System (INIS)

Chorismate mutase, a branch-point enzyme in the aromatic amino acid pathway of Saccharomyces cerevisiae, and also a mutant chorismate mutase with a single amino acid substitution in the C-terminal part of the protein have been purified approximately 20-fold and 64-fold from overproducing strains, respectively. The wild-type enzyme is activated by tryptophan and subject to feedback inhibition by tyrosine, whereas the mutant enzyme does not respond to activation by tryptophan nor inhibition by tyrosine. Both enzymes are dimers consisting of two identical subunits of M_r 30,000, each one capable of binding one substrate and one activator molecule. Each subunit of the wild-type enzyme also binds one inhibitor molecule, whereas the mutant enzyme lost this ability. The enzyme reaction was observed by "1H NMR and shows a direct and irreversible conversion of chorismate to prephenate without the accumulation of any enzyme-free intermediates. The kinetic data of the wild-type chorismate mutase ...

International Nuclear Information System (INIS)

The changes in quality of irradiated spices, i.e., nutmeg (Myristica fragrans), black pepper and white pepper (Piper nigrum) stored in different indigenous packaging materials were evaluated. Packaging materials used were tin cans; tin can lined with polypropylene film, tin can lined with wrapping paper, woven polypropylene bag and woven polypropylene bag lined with polypropylene film. Packaging in woven polypropylene bag could cause significant changes in a_w-values and moisture contents of the spices. In the tin can, no significant change was found during storage. By applying a polypropylene film liner inside the plastic bag, the changes could be retarded. The other parameters observed, i.e. total plate count, total mould and yeast count and organoleptic scores of the samples were not significantly affected by the packaging method. A dose of 5 kGy could reduce the microbial load of the spices by about 2-3 log cycles. A significant decrease in a_w-value and ...

Science.gov (United States)

Indoleamine 2,3-dioxygenase (IDO) and tryptophan 2,3-dioxygenase (TDO) are tryptophan-degrading enzymes. Mammalian IDO expression is induced by cytokines and has antimicrobial and immunomodulatory effects. A major role of mammalian TDO is to supply nicotinamide adenine dinucleotide (NAD(+)). In fungi, the IDO homologue is thought to be expressed constitutively and supply NAD(+), as TDO is absent from their genomes. Here, we reveal the distribution of IDO genes among fungal species and characterize their enzymatic activity. The yeast, Saccharomyces cerevisiae has only one IDO gene, whereas the koji-mold, Aspergillus oryzae has two genes, IDO? and IDO?. The A. oryzae IDO? showed more similar enzymatic properties to those of S. cerevisiae IDO than IDO?, suggesting that the A. oryzae IDO? is a functional homologue of the S. cerevisiae IDO. From the IDO? gene, two isoforms, IDO? and IDO?(+) could be generated by alternative splicing. The latter contained a 17 amino ...

Energy Technology Data Exchange (ETDEWEB)

Numerous studies have investigated degradation of individual endocrine disrupting compounds (EDCs) in lab or natural waters. However, natural variations in water matrices and mixtures of EDCs in the environment may confound analysis of the treatment efficiency. Because chemical based analytical methods cannot represent the combined or synergistic activities between water quality parameters and/or the EDC mixtures at environmentally relevant concentrations ({mu}g L{sup -1}-ng L{sup -1}), bioanalytical assessments of residual estrogenic activity in treated water were used to evaluate the performance of the UV based advanced oxidation process for estrogenic contaminants in water. Four EDCs including estradiol (E{sub 2}), ethinyl estradiol (EE{sub 2}), bisphenol-A (BPA) and nonylphenol (NP) were spiked individually or as a mixture at {mu}g L{sup -1}-ng L{sup -1} in laboratory or natural river water. The removal rates of estrogenic activity were quantitatively evaluated by in vitro ...

Energy Technology Data Exchange (ETDEWEB)

Recent central processing of domestic wastewater in Malaysia has initiated investigations into the disposal/utilization of the sewage sludge produced. We had conducted an experiment to investigate the feasibility of using dewatered sewage sludge and coconut coir as a peat substitute in a potting medium for chrysanthemum. The experiment involved 9 treatments with sewage sludge (SS) and coconut coir (CC) mixed in different ratios (v/v) to replace peat in the standard potting medium of 3:2:1 (soil: peat: sand).The potting medium contained the following treatments, T1: peat + recommended rates of Agroblend (Ag), a slow release fertilizer, and Grofas (Gf), a foliar fertilizer (commonly used medium and fertilization), T2: [1SS:1CC] + Ag, T3: [1SS:1CC] + half recommended rates of Agroblend and Grofas (1/2Ag + 1/2Gf), T4: [2SS:1CC] + Ag, T5: [2SS:1CC] + 1/2Ag + 1/2Gf, T6: [3SS:1CC] + Ag, T7: [3SS:1CC] + 1/2Ag + 1/2Gf, T8: [4SS:1CC] + Ag, and T9: [4SS:1CC] + 1/2Ag + 1/2Gf; laid-out in a ...

Energy Technology Data Exchange (ETDEWEB)

The overall purpose of this project is to evaluate the biological and economic feasibility of restoring high-quality forests on mined land, and to measure carbon sequestration and wood production benefits that would be achieved from forest restoration procedures. In this quarterly report, we present a preliminary comparison of the carbon sequestration potential of forests growing on 14 mined sites in a seven-state region in the Midwestern and Eastern Coalfields. Carbon contents of these forests were compared to adjacent forests on non-mined land. The study was installed as a 3 x 3 factorial in a random complete block design with three replications at each location. The treatments include three forest types (white pine, hybrid poplar, mixed hardwood) and three silvicultural regimes (competition control, competition control plus tillage, competition control plus tillage plus fertilization). Each individual treatment plot is 0.5 acres. Each block of nine plots ...

International Nuclear Information System (INIS)

PAH biomineralization measurements of yardwaste compost samples indicated heterogeneous distribution of active microorganisms and substantial sequestration of the non-polar substrate in the compost matrix. - The purpose of the study was to evaluate the potential of phenanthrene and fluoranthene biodegradation in yardwaste compost materials. These polynuclear aromatic hydrocarbons were chosen for this work because they are relatively readily biodegradable and ubiquitous in the environment. Compost samples were incubated in biometers with "1"4C-labeled phenanthrene and the evolution of "1"4CO_2 was assessed as a measure of mineralization. The "1"4CO_2 evolution varied widely among replicate biometers, possibly as the result of (1) uneven and patchy colonization of phenanthrene-degrading microorganisms on compost particles, and (2) non-uniform dispersion of the labeled substrate spike into the yardwaste microenvironment. Mineralization of phenanthrene reached about ...

International Nuclear Information System (INIS)

The influence of serum, IGF-1 and TGF-#beta# on the differentiation of preadipocytes was examined in primary cultures of porcine adipose tissue cells. In serum-supplemented or serum-free, IGF-1 (1 and 10 nM) had no effect on total cell number. However, IGF-1 (10nM) increased adipocyte number only in serum-supplemented (1% pig serum) cultures, whereas TGF-#beta# (15 pm) reduced the adipocyte number in the presence and absence of IGF-1. Replication of preadipocytes was analyzed with a ["3H] thymidine assay. Preadipocyte proliferation (cpm in adipocyte fraction) was increased by IGF-1 (10nM) only in cultures containing pig serum. TGF-#beta# had no effect on preadipocyte proliferation specifically, but slightly increased total ["3H] thymidine incorporation in cultures with serum. Glycerol phosphate dehydrogenase (GPDH) specific activity was decreased by adding TGF-#beta# to serum-free cultures but TGF-#beta# had little effect in serum-supplemented cultures. Cellular ...

Energy Technology Data Exchange (ETDEWEB)

A transplant experiment was conducted to determine whether standard metabolic rate (SMR) of a freshwater shrimp (Palaeomonetes paludosus) would be affected by exposure to trace element-enriched coal combustion waste (coal ash). Shrimp were transplanted into replicate cages in a coal ash-polluted site and a reference site for 8 months. The coal ash-polluted site was characterized by elevated sediment concentrations of As, Cd, Cr, Cu, Ni, Pb and Se compared to sediments in the reference site. After 8 months in the study sites, shrimp in the polluted site appeared to have accumulated As, Cd and Se from the habitat, but there were on differences in survival between the study sites. However, mean SMR of shrimp (measured as O{sub 2} consumption at rest) held in the polluted site was 51% higher than mean SMR of shrimp held in the reference site. The elevation in SMR indicates that the energetic costs of maintenance are greater for shrimp chronically exposed to the ...

International Nuclear Information System (INIS)

Sodium is one of the most essential elements needed for metabolic processes amongst human beings. It is consumed in the form of sodium chloride but it is also present in edible plant leaves. Sodium is mostly analyzed by flame photometric method, a destructive and time consuming technique. Sodium has been determined in some green leave vegetables samples-palak, radish, khatta palak (ambat chuka), chaulai leaves, chauli bean covers and its seeds by instrumental neutron activation analysis. The method involves irradiation of samples with thermal neutrons from "2"4"1Am-Be source and counting "2"4Na activity (half life 15 hr) from the reaction "2"3Na(n,#gamma#) "2"4Na. Activity due to 1.37 MeV photopeak was counted with a NaI(Tl) crystal coupled to gamma ray spectrometer. Green leaves of the vegetables were thoroughly washed, dried at constant temperature and powdered. Bowen's Kale powder was used as standard for measuring sodium abundances. About 2g each of samples and the standard were ...

Energy Technology Data Exchange (ETDEWEB)

Attention-deficit hyperactivity disorder (ADHD) has been shown to be familial and heritable, in previous studies. As with most psychiatric disorders, examination of pedigrees has not revealed a consistent Mendelian mode of transmission. The response of ADHD patients to medications that inhibit the dopamine transporter, including methylphenidate, amphetamine, pemoline, and bupropion, led us to consider the dopamine transporter as a primary candidate gene for ADHD. To avoid effects of population stratification and to avoid the problem of classification of relatives with other psychiatric disorders as affected or unaffected, we used the haplotype-based haplotype relative risk (HHRR) method to test for association between a VNTR polymorphism at the dopamine transporter locus (DAT1) and DSM-III-R-diagnosed ADHD (N = 49) and undifferentiated attention-deficit disorder (UADD) (N = 8) in trios composed of father, mother, and affected offspring. HHRR analysis revealed significant association ...

Energy Technology Data Exchange (ETDEWEB)

The Tennessee Valley Authority (TVA), the Electric Power Research Institute (EPRI), and the Department of Energy-National Energy Technologies Laboratory (DOE-NETL) are evaluating and demonstrating integration of terrestrial carbon sequestration techniques at a coal-fired electric power plant through the use of Flue Gas Desulfurization (FGD) system gypsum as a soil amendment and mulch, and coal fly ash pond process water for periodic irrigation. From January to March 2002, the Project Team initiated the construction of a 40 ha Carbon Capture and Water Emissions Treatment System (CCWESTRS) near TVA's Paradise Fossil Plant on marginally reclaimed surface coal mine lands in Kentucky. The CCWESTRS is growing commercial grade trees and cover crops and is expected to sequester 1.5-2.0 MT/ha carbon per year over a 20-year period. The concept could be used to meet a portion of the timber industry's needs while simultaneously sequestering carbon in lands which would otherwise ...

Energy Technology Data Exchange (ETDEWEB)

This report forms part of an independent review of the specifications, execution and results of Task 6 of the Aespoe Task Force on Modelling of Groundwater Flow and Transport of Solutes, which is seeking to provide a bridge between site characterization (SC) and performance assessment (PA) approaches to solute transport in fractured rock. The present report is concerned solely with Task 6C, which relates to the construction and parametrisation of a block-scale hydrostructural model of the TRUE Block Scale region of the Aespoe Hard Rock laboratory. The task objectives, specifications and outcome are summarised and reviewed. Also, consideration is given to how the hydrostructural model might affect the outcomes of Task 6D and 6E. The main conclusions of this review are summarised below: The Task 6C hydrostructural model is a more comprehensive approach to quantitatively describing a volume of fractured rock than has been achieved hitherto. The idea of including solute retention ...

Energy Technology Data Exchange (ETDEWEB)

The Integrated Status and Effectiveness Monitoring Program (ISEMP - BPA project No.2003-0017) has been created as a cost effective means of developing protocols and new technologies, novel indicators, sample designs, analytical, data management and communication tools and skills, and restoration experiments that support the development of a region-wide Research, Monitoring and Evaluation (RME) program to assess the status of anadromous salmonid populations, their tributary habitat and restoration and management actions. The most straightforward approach to developing a regional-scale monitoring and evaluation program would be to increase standardization among status and trend monitoring programs. However, the diversity of species and their habitat, as well as the overwhelming uncertainty surrounding indicators, metrics, and data interpretation methods, requires the testing of multiple approaches. Thus, the approach ISEMP has adopted is to develop a broad template that may differ in the ...