Metastases suppressor NME2 associates with telomere ends and telomerase and reduces telomerase activity within cells

Analysis of chromatin-immunoprecipitation followed by sequencing (ChIP-seq) usually disregards sequence reads that do not map within binding positions (peaks). Using an unbiased approach, we analysed all reads, both that mapped and ones that were not included as part of peaks. ChIP-seq experiments were performed in human lung adenocarcinoma and fibrosarcoma cells for the metastasis suppressor non-metastatic 2 (NME2). Surprisingly, we identified sequence reads that uniquely represented human telomere ends in both cases. In vivo presence of NME2 at telomere ends was validated using independent methods and as further evidence we found intranuclear association of NME2 and the telomere repeat binding factor 2. Most remarkably, results demonstrate that NME2 associates with telomerase and reduces telomerase activity in vitro and in vivo, and sustained NME2 expression resulted in reduced telomere length in aggressive human cancer cells. Anti-metastatic function of NME2 has been demonstrated in human cancers, however, mechanisms are poorly understood. Together, findings reported here suggest a novel role for NME2 as a telomere binding protein that can alter telomerase function and telomere length. This presents an opportunity to investigate telomere-related interactions in metastasis suppression.     

Kinetics of medium-chain-length polyhydroxyalkanoate production by a novel isolate of Pseudomonas putida LS46

Six bacteria that synthesize medium-chain-length polyhydroxyalkanoates (mcl-PHAs) were isolated from sewage sludge and hog barn wash and identified as strains of Pseudomonas and Comamonas by 16S rDNA gene sequencing. One isolate, Pseudomonas putida LS46, showed good PHA production (22% of cell dry mass) in glucose medium, and it was selected for further studies. While it is closely related to other P.?putida strains (F1, KT2440, BIRD-1, GB-1, S16, and W619), P.?putida LS46 was genetically distinct from these other strains on the basis of nucleotide sequence analysis of the cpn60 gene hypervariable region. PHA production was detected as early as 12?h in both nitrogen-limited and nitrogen-excess conditions. The increase in PHA production after 48?h was higher in nitrogen-limited cultures than in nitrogen-excess cultures. Pseudomonas?putida LS46 produced mcl-PHAs when cultured with glucose, glycerol, or C(6)-C(14) saturated fatty acids as carbon sources, and mcl-PHAs accounted for 56% of the cell dry mass when cells were batch cultured in medium containing 20?mmol/L octanoate. Although 3-hydroxydecanoate was the major mcl-PHA monomer (58.1-68.8?mol%) in P.?putida LS46 cultured in glucose medium, 3-hydroxyoctanoate was the major monomer produced in octanoate medium (88?mol%).     

Oligonucleotide-stabilized fluorescent silver nanoclusters for turn-on detection of melamine

Fluorescence nanoclusters have been used for the determination of melamine for the first time. The method is based on the fluorescence turn-on of oligonucleotide-stabilized silver nanoclusters (DNA-Ag NCs) by melamine. The enhancement factors (I-I(0))/I(0) increase linearly with melamine concentrations over the range 5.0×10(-8)-7.0×10(-6) M (R(2)=0.998). The detection limit is 1.0×10(-8) M, which is approximately 2000 times lower than the US Food and Drug Administration estimated melamine safety limit of 20.0 μM. Furthermore, the milk samples spiked with melamine are analyzed with excellent recoveries.     

Benzimidazole bearing oxadiazole and triazolo-thiadiazoles nucleus: Design and synthesis as anticancer agents

Two new series of benzimidazole bearing oxadiazole[1-(1H-benzo[d]imidazol-2-yl)-3-(5-substituted-1,3,4-oxadiazol-2-yl)propan-1-ones (4a-l)] and triazolo-thiadiazoles[1-(1H-benzo[d]imidazol-2-yl)-3-(6-(substituted)-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazol-3-yl)propan-1-one (7a-e)] have been synthesized successfully from4-(1H-benzo[d]imidazol-2-yl)-4-oxobutanehydrazide (3) with an aim to produce promising anticancer agents. In vitro anticancer activities of synthesized compounds were screened at the National Cancer Institute (NCI), USA, according to their applied protocol against full NCI 60 human cell lines panel; results showed good to remarkable anticancer activity. Among them, compound (4j, NCS: 761980) exhibited significant growth inhibition and further screened at 10-fold dilutions of five different concentrations (0.01, 0.1, 1, 10 and 100μM) with GI(50) values ranging from 0.49 to 48.0μM and found superior for the non-small cell lung cancer cell lines like HOP-92 (GI(50) 0.49, TGI 19.9,LC(50) >100 and Log(10)GI(50) -6.30, Log(10)TGI -4.70, Log(10)LC(50) >-4.00).     

D-Aspartate oxidase and D-amino acid oxidase are localised in the peroxisomes of terrestrial gastropods

D-Aspartate oxidase and D-amino acid oxidase were found in high activity in the tissues of representative species of terrestrial gastropods. Analytical subcellular fractionation demonstrated that both of these oxidases co-localised with the peroxisome markers, acyl-CoA oxidase and catalase, in the digestive gland homogenate. Electron microscopy of peak peroxisome fractions showed particles of uniform size with generally well preserved variably electron-dense matrices bounded by an apparently single limiting membrane. Many of the particles exhibited a core region of enhanced electron density. Catalase cytochemistry of peak fractions confirmed the peroxisome identity of the organelles. Peroxisome-enriched subcellular fractions were used to investigate the properties of gastropod D-aspartate oxidase and D-amino acid oxidase activities. The substrate and inhibitor specificities of the two activities demonstrated that two distinct enzymes were present analogous to, but not identical to, the equivalent mammalian peroxisomal enzymes.     

Microsatellite Marker Linked to a Leaf Rust Resistance Gene from Triticum monococcum L Transferred to Bread Wheat

Triticum monococcum L, a diploid wheat species closely related to the A genome of cultivated wheats, is highly resistant to leaf rust. A synthetic amphiploid, T. monococcum — T. durum was crossed with T. aestivum cv WL711, highly susceptible to leaf rust. Leaf rust resistant derivatives were selected among backcross generations with the recurrent parent WL711 and cytologically analysed. Chromosome number of the leaf rust resistant BC₁F₃ progenies varied from 39 to 44. Six leaf rust resistant and susceptible bulks from different BC₁F₃ progenies were analysed using 29 wheat microsatellite(WMS) markers already mapped on A genome of bread wheat and found polymorphic among parents. One T. monococcum specific allele of WMS gwm136 locus was found to be closely linked to the leaf rust resistance gene in all the resistant bulks. Differential chromosome number, frequency of univalents and multivalents, however, indicated that the critical T. monococcum chromosome might be present in addition to the A genome chromosomes of wheat, substituted either for the B or D genome chromosome of wheat or translocated to chromosome 1A of wheat in one or the other bulks. The association of the T. monococcum specific allele of WMS gwm136 locus to leaf rust resistance was further confirmed from bulked segregant analysis in BC₂F₁ generation.     

Pro-oxidant, anti-oxidant and cleavage activities on DNA of curcumin and its derivatives demethoxycurcumin and bisdemethoxycurcumin.

Curcumin, a naturally occurring phytochemical responsible for the colour of turmeric shows a wide range of pharmacological properties including antioxidant, anti-inflammatory and anti-cancer effects. We have earlier shown that curcumin in the presence of Cu(II) causes strand cleavage in DNA through generation of reactive oxygen species, particularly the hydroxyl radical. Thus, curcumin shows both antioxidant as well as pro-oxidant effects. In order to understand the chemical basis of various biological properties of curcumin, we have studied the structure-activity relationship between curcumin and its two naturally occurring derivatives namely demethoxycurcumin (dmC) and bisdemethoxycurcumin (bdmC). Curcumin was found to be the most effective in the DNA cleavage reaction and a reducer of Cu(II) followed by dmC and bdmC. The rate of formation of hydroxyl radicals by the three curcuminoids also showed a similar pattern. The relative antioxidant activity was examined by studying the effect of these curcuminoids on cleavage of plasmid DNA by Fe(II)-EDTA system (hydroxyl radicals) and the generation of singlet oxygen by riboflavin. The results indicate that curcumin is considerably more active both as an antioxidant as well as an oxidative DNA cleaving agent. The DNA cleavage activity is the consequence of binding of Cu(II) to various sites on the curcumin molecule. Based on the present results, we propose three binding sites for Cu(II). Two of the sites are provided by the phenolic and methoxy groups on the two benzene rings and the third site is due to the presence of 1,3-diketone system between the rings. Furthermore, both the antioxidant as well as pro-oxidant effects of curcuminoids are determined by the same structural moieties.     

Fibre Optic SPR Sensor Using Functionalized CNTs for the Detection of SMX: Comparison with Enzymatic Approach

Plasmonics - A novel approach for the detection of sulfamethaxazole (SMX) based on fibre optic surface plasmon resonance (SPR) technique and functionalized carbon nanotubes (CNTs) is proposed for 0...     

Shortening of sleep length and delayed mid-sleep on free days are the characteristic features of predominantly morning active population of Indian teenagers

Sleep and Biological Rhythms - We studied patterns in the distribution of chronotype and sleep–wake behavior in a randomly chosen sample population of Indian teenagers consisting 965...