Stress-induced mutagenesis in bacteria

The evolutionary significance of stress-induced mutagenesis was evaluated by studying mutagenesis in aging colonies (MAC) of Escherichia coli natural isolates. A large fraction of isolates exhibited a strong MAC, and the high MAC variability reflected the diversity of selective pressures in ecological niches. MAC depends on starvation, oxygen, and RpoS and adenosine 3',5'-monophosphate regulons; thus it may be a by-product of genetic strategies for improving survival under stress. MAC could also be selected through beneficial mutations that it generates, as shown by computer modeling and the patterns of stress-inducible and constitutive mutagenesis. We suggest that irrespective of the causes of their emergence, stress-induced mutations participate in adaptive evolution.     

Phylogenetic shadowing of primate sequences to find functional regions of the human genome

Nonhuman primates represent the most relevant model organisms to understand the biology of Homo sapiens. The recent divergence and associated overall sequence conservation between individual members of this taxon have nonetheless largely precluded the use of primates in comparative sequence studies. We used sequence comparisons of an extensive set of Old World and New World monkeys and hominoids to identify functional regions in the human genome. Analysis of these data enabled the discovery of primate-specific gene regulatory elements and the demarcation of the exons of multiple genes. Much of the information content of the comprehensive primate sequence comparisons could be captured with a small subset of phylogenetically close primates. These results demonstrate the utility of intraprimate sequence comparisons to discover common mammalian as well as primate-specific functional elements in the human genome, which are unattainable through the evaluation of more evolutionarily distant species.     

Electron transfer-induced dynamics of oxygen molecules on the TiO2(110) surface

Diffusion of oxygen molecules on transition metal oxide surfaces plays a vital role for the understanding of catalysis and photocatalysis on these materials. By means of time-resolved scanning tunneling microscopy, we provide evidence for a charge transfer-induced diffusion mechanism for O2 molecules adsorbed on a rutile TiO2(110) surface. The O2 hopping rate depended on the number of surface donors (oxygen vacancies), which determines the density of conduction band electrons. These results may have implications for the understanding of oxidation processes on metal oxides in general.     

Proteomic profile of pre-implantational ovine embryos produced in vivo

The present study was conducted to decipher the proteome of in vivo-produced pre-implantation ovine embryos. Ten locally adapted Morana Nova ewes received hormonal treatment and were inseminated 12 hr after ovulation. Six days later, 54 embryos (morula and blastocyst developmental state) were recovered from eight ewes and pooled to obtain sufficient protein for proteomic analysis. Extracted embryo proteins were analysed by LC-MS/MS, followed by identification based on four database searches (PEAKS, Proteome Discoverer software, SearchGUI software, PepExplorer). Identified proteins were analysed for gene ontology terms, protein clusters and interactions. Genes associated with the ovine embryo proteome were screened for miRNA targets using data sets of TargetScan ( http://www.targetscan.org ) and mIRBase ( http://www.mirbase.org ) servers. There were 667 proteins identified in the ovine embryos. Biological processes of such proteins were mainly related to cellular process and regulation, and molecular functions, to binding and catalytic activity. Analysis of the embryo proteins revealed 49 enriched functional clusters, linked to energy metabolism (TCA cycle, pyruvate and glycolysis metabolism), zona pellucida (ZP), MAPK signalling pathway, tight junction, binding of sperm to ZP, translation, proteasome, cell cycle and calcium/phospholipid binding. Sixteen miRNAs were related to 25 pre-implantation ovine embryo genes, all conserved in human, bovine and ovine species. The interaction network generated by miRNet showed four key miRNAs (hsa-mir-106b-5p; hsa-mir-30-5p; hsa-mir-103a-5p and hsa-mir-106a-5p) with potential interactions with embryo-expressed genes. Functional analysis of the network indicated that miRNAs modulate genes related to cell cycle, regulation of stem cell and embryonic cell differentiation, among others. Retrieved miRNAs also modulate the expression of genes involved in cell signalling pathways, such as MAPK, Wnt, TGF-beta, p53 and Toll-like receptor. The current study describes the first major proteomic profile of 6-day-old ovine embryos produced in vivo, setting a comprehensive foundation for our understanding of embryo physiology in the ovine species.     

Distribution and relative abundance of peccaries in the Argentine Chaco: associations with human factors

In the Argentine Chaco, the three species of peccaries (Artiodactyla) are likely affected by habitat destruction and hunting, yet basic information on peccary distribution and status in this region is poorly known. This study identifies human factors associated with relative abundance and distribution of each species. Estimation of relative abundance was based on interviews with local hunters, and variables potentially related to distribution and relative abundance of peccaries were estimated for 153 circular sample sites of 10 km diameter in the Argentine Chaco. Peccaries were found in sites of high forest cover, low human density, far from towns and with low density of roads. After accounting for forest cover and other variables, number of settlements was identified as the main variable negatively associated with relative abundance of the three species, which may be a result of local hunting. Density of roads was also negatively associated with presence of chacoan peccaries. Collared peccary seems to be the least susceptible to human perturbations. It was the most widely distributed and found in a wider range of conditions than the other species. Chacoan peccary was relatively rare. Because colonization and development programs are increasing in the region, areas still uninhabited should be protected, construction of roads controlled, and hunting managed.     

Selenium supplementation can protect cultured rat cardiomyocytes from hypoxia/reoxygenation damage

The possibility of enhancing glutathione peroxidase (GPx) activity and cytosolic total antioxidant activity (TAA) in normoxia and hypoxia/reoxygenation (H/R) by the supplementation of different concentrations of sodium selenite (SS) or selenomethionine (SM) was investigated in cultured rat cardiomyocytes. To assess the entity of oxidative stress due to H/R, levels of conjugated dienes containing lipids were determined. In normoxia, GPx activity and TAA increased in parallel with the increase in SS and SM supplementation. H/R did not influence GPx activity but lowered TAA; both SS and SM supplementations were effective in increasing GPx activity, the most effective concentration being 1 microM. At this SS and SM concentration, TAA returned to a normoxic value. Conjugated diene production, increased by H/R, was reduced by SS and SM supplementation, the 1 microM concentration appearing to be the most effective one. According to these data Se supplementation represents another possibility to counteract oxidative damage in the myocardium.     

A rapid aflatoxin B1 ELISA: development and validation with reduced matrix effects for peanuts, corn, pistachio, and Soybeans

Among the competitive ELISAs for aflatoxins that have been described, few have been adequately validated for reduced matrix effects. Using an aflatoxin B(1) (AFB(1))-specific polyclonal antibody (produced from AFB(1)-oxime conjugated to bovine serum albumin (BSA)) and AFB(1)- and AFB(2)-enzyme conjugates, four direct competitive ELISAs based on 96-microwell plates (two standard assays and two rapid assays) were developed, paying special attention to producing a robust assay relatively free of interferences for a range of agricultural products. The antibody was AFB(1)-specific, detecting only AFB(1) in a mixture of four aflatoxins (AFB(1), AFB(2), AFG(1), and AFG(2)), but showed significant cross-reaction with AFG(1) (57-61%) when an individual compound was tested. Standard assays (long assays) exhibited higher sensitivities than rapid assays (short assays) with IC(50) values of 12 +/- 1.5 and 9 +/- 1.5 microg/kg in sample (with 1 in 5 dilution of sample extract) for AFB(1) and AFB(2)-enzyme conjugates, respectively. These assays have narrower detection ranges (7.1-55.5 microg/kg in sample) and required dilution of sample extracts to overcome solvent and matrix interferences, making these assays less ideal as analytical methods. Rapid assays exhibited IC(50) values of 21.6 +/- 2.7 and 12 microg/kg in sample for AFB(1)- and AFB(2)-enzyme conjugates, respectively. These assays have ideally broader detection ranges (4.2-99.9 microg/kg in sample) and showed no methanol effects up to 80% with significantly reduced matrix interferences as a result of the shorter incubation times and increasing the amounts of enzyme conjugate used. Therefore, the rapid assays were formatted to perform without a need for extract dilution. The rapid assays can be completed within 15 min, potentially suitable for receival bays where quick decision-making to segregate low and high contamination is critical. Further validation using the rapid assay with AFB(1)-enzyme conjugate indicated relatively good recoveries of AFB(1) spiked in corn, peanuts, pistachio, and soybeans, which were free from significant matrix effects. It can be concluded that this rapid assay would be suitable for monitoring aflatoxin AFB(1) at current legal maximum residue limits of 10 microg/kg in food such as corn, peanuts, pistachio, and soybeans.     

Biotransformation of an organochlorine insecticide,endosulfan, by Anabaena species

This study assesses the role of the blue-green algal species present in the soil in the dissipation of endosulfan and its metabolites in the soil environment. Two Anabaena species, Anabaena sp. PCC 7120 and Anabaena flos-aquae, were used in this study. Anabaena sp. PCC 7120 produced three principal biotransformation compounds, chiefly endosulfan diol (endodiol), and minor amounts of endosulfan hydroxyether and endosulfan lactone. Trace amounts of endosulfan sulfate were detected. In comparison, the biotransformation of endosulfan by Anabaena flos-aquae yielded mainly endodiol with minor amounts of endosulfan sulfate. An unknown compound was produced up to 70% from endosulfan spiked in the medium inoculated by A. flos-aquae after 8 days of incubation. Therefore, the endosulfan fate was dependent on the species. Within 1 day of incubation, two Anabaena species produced low amounts of beta-endosulfan after application of alpha-endosulfan. These results suggest the presence of isomerase in the Anabaena species. Further studies using a fermentor to control the medium pH at 7.2 to minimize chemical hydrolysis of endosulfan revealed a major production of endodiol with minor amounts of endosulfan sulfate and the unknown compound. These results showed that the production of the unknown compound might be dependent on the alkaline pH in the medium and that the production of endodiol by A. flos-aquae might be biologically controlled. This study showed that two algal species could contribute in the detoxification pathways of endosulfan in the soil environment.     

Two microsatellite markers flanking a dominant gene for resistance to soybean cust nematode race 3

The purpose of this work was to identifymicrosatellite markers linked to a gene forresistance to Heterodera glycinesIchinohe (Soybean Cyst Nematode – SCN) insoybean cultivar Hartwig. ABC₁F₂ mapping population derivedfrom a cross between Hartwig (resistant)and the Brazilian soybean line Y23(susceptible) was used. About 200microsatellite or simple sequence repeat(SSR) primer pairs were tested in a bulkedsegregant analysis (BSA). Those thatshowed clear polymorphisms were amplifiedin the BC₁F₂ population, whichhad been previously inoculated andevaluated for resistance/susceptibility toSCN Race 3. Three SSR markers linked toSCN resistance were detected in thepopulation. Two of them, Satt 038 and Satt163, flanking a dominant resistant gene(d/a = –0.90), explained 37% of thephenotypic variance. This gene was mappedat the edge of molecular linkage group G. Broad and narrow sense heritabilities wereestimated to be 50.54% and 37.73%,respectively. A selection efficiency of91.18% was obtained with the simultaneoususe of the two markers. The identified SSRmarkers will be useful tools for assistingthe selection of homozygous genotypes andfor expediting the introgression of the SCNresistance locus from cv. Hartwig tosoybean elite cultivars.     

Fertility restoration and nor suppression caused by Aegilops mutica chromosomes in alloplasmic hybrids and lines

By crossing Aegilops mutica with Triticum dicoccum as a bridge species and backcrossing with common wheat as a recurrent pollen parent, male sterile alloplasmic line(s) were produced. In progeny of the crosses, a self fertile plant with 42 chromosomes was selected and named R 20. From this plant several lines that possessed Rf (fertility restoring) genes and/or powdery mildew resistant genes were obtained. Apparently, the system of sterility-fertility of pollen can be applied for hybrid wheat production. In addition, the disease resistance may be used in breeding. The male fertile lines possessed one or more Ae. mutica sat-chromosome(s), which show the ability to suppress the nucleolar organizing regions of chromosomes 1B and 6B of common wheat. The relation between the sat-chromosomes and male fertility restoration is not yet clear. This revised version was published online in July 2006 with corrections to the Cover Date.