Identification of a DNA nonhomologous end-joining complex in bacteria

In eukaryotic cells, double-strand breaks (DSBs) in DNA are generally repaired by the pathway of homologous recombination or by DNA nonhomologous end joining (NHEJ). Both pathways have been highly conserved throughout eukaryotic evolution, but no equivalent NHEJ system has been identified in prokaryotes. The NHEJ pathway requires a DNA end-binding component called Ku. We have identified bacterial Ku homologs and show that these proteins retain the biochemical characteristics of the eukaryotic Ku heterodimer. Furthermore, we show that bacterial Ku specifically recruits DNA ligase to DNA ends and stimulates DNA ligation. Loss of these proteins leads to hypersensitivity to ionizing radiation in Bacillus subtilis. These data provide evidence that many bacteria possess a DNA DSB repair apparatus that shares many features with the NHEJ system of eukarya and suggest that this DNA repair pathway arose before the prokaryotic and eukaryotic lineages diverged.     

Regulation of cytokine receptors by Golgi N-glycan processing and endocytosis

The Golgi enzyme beta1,6 N-acetylglucosaminyltransferase V (Mgat5) is up-regulated in carcinomas and promotes the substitution of N-glycan with poly N-acetyllactosamine, the preferred ligand for galectin-3 (Gal-3). Here, we report that expression of Mgat5 sensitized mouse cells to multiple cytokines. Gal-3 cross-linked Mgat5-modified N-glycans on epidermal growth factor and transforming growth factor-beta receptors at the cell surface and delayed their removal by constitutive endocytosis. Mgat5 expression in mammary carcinoma was rate limiting for cytokine signaling and consequently for epithelial-mesenchymal transition, cell motility, and tumor metastasis. Mgat5 also promoted cytokine-mediated leukocyte signaling, phagocytosis, and extravasation in vivo. Thus, conditional regulation of N-glycan processing drives synchronous modification of cytokine receptors, which balances their surface retention against loss via endocytosis.     

Soil microbial communities resistant to changes in plant functional group composition

The soil community is an often ignored part of research which links plant biodiversity and ecosystem functioning despite their influence on numerous functions such as decomposition and nutrient cycling. Few consistent patterns have been detected that link plant and soil community composition. We used a removal experiment in a northern Canadian grassland to examine the effects of plant functional group identity on soil microbial community structure and function. Plant functional groups (graminoids, legumes and forbs) were removed independently from plots for five growing seasons (2003-2007) and in the fifth year effects on the soil microbial community were examined using substrate-induced respiration (SIR - a measure of metabolic diversity) and phospholipid fatty acid analysis (PLFA - a measure of microbial community composition). Removal treatments were also crossed with both a fertilizer treatment and a fungicide treatment to determine if effects of functional group identity on the soil community were context dependent. Plant functional group identity had almost no effect on the soil microbial community as measured by either SIR or PLFA. Likewise, soil properties including total carbon, pH, moisture and nutrients showed a limited response to plant removals in the fifth year after removals. We found a direct effect of fertilizer on the soil community, with fertilized plots having decreased metabolic diversity, with a decreased ability to metabolize amino acids and a phenolic acid, but there was no direct soil microbial response to fungicide. We show that in this northern Canadian grassland the soil microbial community is relatively insensitive to changes in plant functional group composition, and suggest that in northern ecosystems, where plant material is only slowly incorporated into the soil, five growing seasons may be insufficient to detect the impact of a changing plant community on the soil microbes.     

Uncommon associations in target resistance among French populations of Myzus persicae from oilseed rape crops

Within the framework of a molecular exploration of target resistance in populations of Myzus persicae on oilseed rapes in France, (1) the S431F mutation (coding gene ace2), although previously reckoned to be rare, revealed to be frequent, (2) M918L (phenotypically characterised) and L932F (both on para) were found for the first time in M. persicae, and (3) a linkage was revealed between M918L and S431F. While until recently populations developing on French oilseed rapes were dominated by genotypes possessing pyrethroid target resistance and esterase overproduction, to date a different type of dominating genotype, equipped with carbamate and pyrethroid target resistance, seems to be invading such fields.     

Ion-pair reverse phase liquid chromatographic determination of sodium acifluorfen in feed

Ion-pair reverse phase liquid chromatography (LC) and UV detection at 280 nm have been used to determine sodium acifluorfen (sodium-5-[2-chloro-4-(trifluoromethyl)-phenoxy]-2-nitrobenzoate), an experimental diphenyl ether herbicide, in dog feed. Sodium-5-(2,4-dichlorophenoxy)-2-nitrobenzoate is used as the internal standard. The feed is homogenized in 0.01N HCl, followed by ethyl acetate extraction, and centrifugation. The organic layer is removed and evaporated and the residue is reconstituted in methanol and filtered before LC analysis (mobile phase methanol-water (58 + 42), 0.005M in tetrabutylammonium phosphate and 0.045M in (NH4)2HPO4, at pH 7.4). The ion-pair technique offers a high degree of control over the retention characteristics of the herbicide and internal standard. The use of the internal standard permits precise and accurate quantitation and substantially reduces analysis time compared with the external standard method.     

Isozyme Banding Pattern and Estimation of Genetic Diversity among Guinea Grass Germplasm

Isozyme banding pattern was studied in Guinea grass (Panicum maximum Jacq.), a widely cultivated grass having good fodder value. Similarity among 63 accessions collected from diverse sources was worked out using five enzyme systems (SOD, GOT, ACP, Esterase and Peroxidase) following horizontal starch gel electrophoresis. Biochemical markers such as isozymes are useful supplements in identifying the genetic variation present in any crop. A total of 35 clear and unambiguous bands were used for analysis of which 8 bands were monomorphic. Polymorphism exhibited by 27 bands from all five enzyme systems indicate presence of considerable diversity in this species. The dendrogram generated after UPGMA and SAHN cluster analysis using Jaccard genetic distance showed that 63 accessions from diverse geographical locations could be grouped in three main clusters, of which two could further be divided into sub-clusters. Although the clusters comprised members from different locations, most of the accessions from similar geographical locations tended to cluster in same group.     

Determination of a chemical basis of air-pollution stress in wood of mature white pine trees in the susceptive forest ecosystems

This paper describes a pilot investigation dealing with changes in chemical composition of wood in pollution-stressed white pine trees, taken from two forested ecosystems of Southern Ontario, Canada. The implications of this research lie in a much needed search for a potential bloindicator of a pre-decline, incipient-stress induced (and/or contributed by) the air-pollutants.     

δ34S variations in vegetation and soil exposed to intense biogenic sulphide emissions near Paige mountain,N.W.T., Canada

Sulphur isotope abundances demonstrate that natural emissions of biogenic H₂S and its oxidation products from springs near Paige Mountain, N.W.T., Canada, can be incorporated into surrounding soil and vegetation. δ³⁴S values as low as ? 33‰ for soil and vegetation are the result of dominant uptake of biogenic atmospheric compounds. In contrast, vegetation on a gypsum outcrop remote from the springs, has δ³⁴S values as high as + 26‰ indicating nearly exclusive derivation of S from the soil. Near the springs, soil with vegetative cover is less depleted in ³⁴S than soil lacking cover. Lower needles on a Black Spruce were found to be less depleted in ³⁴S than the upper needles. These observations suggest that upper foliage exerts a canopy effect on lower foliage and in turn, interception by vegetation reduces the flux of atmospheric S compounds to the soil. Clearly, natural emissions of S compounds can interfere in studies of long range transport of industrial emissions; S isotope analyses might identify such interferences and reduce the chance of misinterpretation.