Mutagenesis by hydrogen peroxide treatment of mammalian cells: a molecular analysis

Hydrogen peroxide is an oxidizing agent which can be generated intracellularly either during normal metabolism or by treatment with external agents including solar UV radiation. Simian cells (CV-1) transfected with the SV40-based shuttle vector plasmid pZ189 have been treated with H2O2 and then incubated to allow repair and replication of the plasmid. The frequency of mutations at the supF locus of the recovered plasmid increases by a factor of up to four over the spontaneous value. The nucleotide changes associated with 100 spontaneous and 100 H2O2-induced mutants have been determined directly by sequencing a 150 bp fragment that includes the entire supF tRNA coding region. Deletions were observed in ∼45% of both the spontaneous and induced mutants, whereas single or multiple base changes arose in 68 and 57% of the induced and spontaneous mutants respectively. The spectrum of induced mutations is characterized by (i) the occurrence of deletions associated with base changes (16% of all mutants analysed) and (ii) small deletions of 3 bp and less (51% of all deletion mutants sequenced). Sixty-five per cent (15 out of 23) of all small deletions (spontaneous and induced) are associated with runs of between two and five identical bases and eight of them arise at a mutational ‘hotspot' region of five cytosines between bp 172 and 176. The majority (19 out of 30) of completely sequenced deletions observed in the spontaneous spectrum contain either (i) small (2-10 bp) direct repeat sequences that lie immediately outside one deletion terminus and immediately inside the second deletion terminus or (ii) small (2-3 bp) inverted repeat sequences lying immediately inside the two deletion termini. Most deletions that we have observed are therefore likely to arise as a consequence of specific aspects of DNA structur

Polarimetric Alos/Palsar-2 data for retrieving aboveground biomass of secondary forest in the Brazilian Amazon

Secondary forests (SFs) are one of the major carbon sink in the Neotropics due to the rapid carbon assimilating in their aboveground biomass (AGB). However, the accurate contribution of the SFs to the carbon cycle is a great challenge because of the uncertainty in AGB estimates. In this context, the main objective of this work is to explore polarimetric Alos/Palsar-2 data from to model AGB in the SFs of the Central Amazon, Amazonas State. Forest inventory was conducted in 2014 with the measured of 23 field plots. Multiple linear regression analysis was performed to select the best model by corrected AIC w and validated by leave-one-out bootstrapping method. The best fitted model has six parameters and explained 65% of the aboveground biomass variability. The prediction error was calculated to be RMSEP = 8.8 ± 2.98 Mg.ha -1 (8.75%). The main polarimetric attributes in the model were those direct related to multiple scattering mechanisms as the Shannon Entropy and the volumetric mechanism of Bhattacharya decomposition; and those related to increase in double-bounce as the co-polarization ratio (VV/HH) resulted of soil-trunk interactions. Such models are intended to improve accuracy for mapping SFs AGB in often cloudy environments as in the Brazilian Amazon

Retrieving secondary forest aboveground biomass from polarimetric ALOS-2 PALSAR-2 data in the Brazilian Amazon

Secondary forests (SF) are important carbon sinks, removing CO2 from the atmosphere through the photosynthesis process and storing photosynthates in their aboveground live biomass (AGB). This process occurring at large-scales partially counteracts C emissions from land-use change, playing, hence, an important role in the global carbon cycle. The absorption rates of carbon in these forests depend on forest physiology, controlled by environmental and climatic conditions, as well as on the past land use, which is rarely considered for retrieving AGB from remotely sensed data. In this context, the main goal of this study is to evaluate the potential of polarimetric (quad-pol) ALOS-2 PALSAR-2 data for estimating AGB in a SF area. Land-use was assessed through Landsat time-series to extract the SF age, period of active land-use (PALU), and frequency of clear cuts (FC) to randomly select the SF plots. A chronosequence of 42 SF plots ranging 3-28 years (20 ha) near the Tapajós National Forest in Pará state was surveyed to quantifying AGB growth. The quad-pol data was explored by testing two regression methods, including non-linear (NL) and multiple linear regression models (MLR).We also evaluated the influence of the past land-use in the retrieving AGB through correlation analysis. The results showed that the biophysical variables were positively correlated with the volumetric scattering, meaning that SF areas presented greater volumetric scattering contribution with increasing forest age. Mean diameter, mean tree height, basal area, species density, and AGB were significant and had the highest Pearson coefficients with the Cloude decomposition (λ3), which in turn, refers to the volumetric contribution backscattering from cross-polarization (HV) (ρ = 0.57-0.66, p-value < 0.001). On the other hand, the historical use (PALU and FC) showed the highest correlation with angular decompositions, being the Touzi target phase angle the highest correlation (Φs) (ρ = 0.37 and ρ = 0.38, respectively). The combination of multiple prediction variables with MLR improved the AGB estimation by 70% comparing to the NL model (R2 adj. = 0.51; RMSE = 38.7 Mg ha-1) bias = 2.1 ± 37.9 Mg ha-1 by incorporate the angular decompositions, related to historical use, and the contribution volumetric scattering, related to forest structure, in the model. The MLR uses six variables, whose selected polarimetric attributes were strongly related with different structural parameters such as the mean forest diameter, basal area, and the mean forest tree height, and not with the AGB as was expected. The uncertainty was estimated to be 18.6% considered all methodological steps of the MLR model. This approach helped us to better understand the relationship between parameters derived from SAR data and the forest structure and its relation to the growth of the secondary forest after deforestation events

Altered glucose homeostasis and hepatic function in obese mice deficient for both kinin receptor genes

The Kallikrein-Kinin System (KKS) has been implicated in several aspects of metabolism, including the regulation of glucose homeostasis and adiposity. Kinins and des-Arg-kinins are the major effectors of this system and promote their effects by binding to two different receptors, the kinin B2 and B1 receptors, respectively. To understand the influence of the KKS on the pathophysiology of obesity and type 2 diabetes (T2DM), we generated an animal model deficient for both kinin receptor genes and leptin (obB1B2KO). Six-month-old obB1B2KO mice showed increased blood glucose levels. Isolated islets of the transgenic animals were more responsive to glucose stimulation releasing greater amounts of insulin, mainly in 3-month-old mice, which was corroborated by elevated serum C-peptide concentrations. Furthermore, they presented hepatomegaly, pronounced steatosis, and increased levels of circulating transaminases. This mouse also demonstrated exacerbated gluconeogenesis during the pyruvate challenge test. The hepatic abnormalities were accompanied by changes in the gene expression of factors linked to glucose and lipid metabolisms in the liver. Thus, we conclude that kinin receptors are important for modulation of insulin secretion and for the preservation of normal glucose levels and hepatic functions in obese mice, suggesting a protective role of the KKS regarding complications associated with obesity and T2DM

Kinase Inhibitor Profile For Human Nek1, Nek6, And Nek7 And Analysis Of The Structural Basis For Inhibitor Specificity

Human Neks are a conserved protein kinase family related to cell cycle progression and cell division and are considered potential drug targets for the treatment of cancer and other pathologies. We screened the activation loop mutant kinases hNek1 and hNek2, wild-type hNek7, and five hNek6 variants in different activation/phosphorylation statesand compared them against 85 compounds using thermal shift denaturation. We identified three compounds with significant Tm shifts: JNK Inhibitor II for hNek1(Ä262-1258)-(T162A), Isogranulatimide for hNek6(S206A), and GSK-3 Inhibitor XIII for hNek7wt. Each one of these compounds was also validated by reducing the kinases activity by at least 25%. The binding sites for these compounds were identified by in silico docking at the ATP-binding site of the respective hNeks. Potential inhibitors were first screened by thermal shift assays, had their efficiency tested by a kinase assay, and were finally analyzed by molecular docking. 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Search for charginos in e+e- interactions at sqrt(s) = 189 GeV

An update of the searches for charginos and gravitinos is presented, based on a data sample corresponding to the 158 pb^{-1} recorded by the DELPHI detector in 1998, at a centre-of-mass energy of 189 GeV. No evidence for a signal was found. The lower mass limits are 4-5 GeV/c^2 higher than those obtained at a centre-of-mass energy of 183 GeV. The (\mu,M_2) MSSM domain excluded by combining the chargino searches with neutralino searches at the Z resonance implies a limit on the mass of the lightest neutralino which, for a heavy sneutrino, is constrained to be above 31.0 GeV/c^2 for tan(beta) \geq 1.Comment: 22 pages, 8 figure