Urea hydrolysis in oxisols: Effects of substrate concentration, temperature, pH, incubation time and storage conditions

This study evaluated the kinetics of urease hydrolysis in two different Brazilian soils (Rhodic Oxisols): a typic alumino-ferric Red Latossol and a typic distrophic Red Latosol. The trials were carried out to study the effects of air drying, soil sampling, storage conditions and temperature, pH, time of incubation, and substrate concentration on the urease activity. Results showed that best conditions for soil sample storage after air-drying for a 7-day period were at room temperature or at a temperature of 5 degrees C. For the studied conditions the best incubation time was one hour at 25-30 degrees C, without pH buffering. The ideal urea concentration suggested for the detection of the maximum velocity was 3.3 g L-1 for the typic alumino-ferric Red Latossol and 2.5 g L-1 for the typic distrophic Red Latosol.29465165

PO-078 Role of the ER stress-autophagy axis and mitochondrial metabolism reprogramming in the apoptosis induced by δ-TOCOTRIENOL in prostate cancer

Introduction Castration resistant prostate cancer (CRPC) is an aggressive tumour with still limited therapeutic outcomes. Tocotrienols (TT), vitamin E derivatives, were reported to exert anticancer activity in different tumours. The aim of this study was to assess the effects of δ-TT on human CRPC cells growth and the molecular mechanisms associated with its activity. Material and methods In human normal prostate (RWPE-1) and CRPC (PC3 and DU145) cell lines the effect of δ-TT on cell viability was evaluated by MTT assay; in PC3 and DU145 cells Trypan blue exclusion and colony formation assays were also performed. The expression of apoptosis-, ER stress- and autophagy-related proteins was analysed by Western blot and immunofluorescence assays, and the cytotoxic effect of δ-TT was also assessed using specific inhibitors of these pathways. The effect on mitochondrial metabolism was evaluated analysing the expression of the OXPHOS complexes (Western blot), the mitochondrial activity and mass (flow cytometry), the oxygen consumption (Clark-type oxygen electrode) and the ATP production (colorimetric assay). Results and discussions We demonstrated that δ-TT exerts a cytotoxic effect on PC3 and DU145 but not on RWPE-1 cells. In particular, δ-TT induces caspase 3 and PARP cleavage and cytochrome c release from mitochondria, and its cytotoxic effect is partially blocked by co-treatment with the pan-caspase inhibitor z-VAD-FMK, confirming that δ-TT exerts a pro-apoptotic effect on CRPC cells. We also observed that δ-TT significantly increases the expression of ER stress (BiP, IRE1α, PERK, pEIF2α, ATF4 and CHOP) and autophagy mediators (LC3-II and p62). Using the ER stress inhibitors salubrinal and 4-phenylbutyrate (4-PBA) and the autophagic flux inhibitors 3-methyladenine and chloroquine, we confirmed that the effect of δ-TT is mediated by both these mechanisms. In addition, treatment with salubrinal or 4-PBA impairs δ-TT-induced LC3-II expression, demonstrating that this compound triggers the ER stress-autophagy axis. Finally, we observed that δ-TT severely alters mitochondrial metabolism: the expression of the OXPHOS protein complexes, the mitochondrial activity/mass ratio, the oxygen consumption and the ATP production were significantly reduced after δ-TT treatment. Conclusion These results demonstrate that δ-TT exerts a selective pro-apoptotic effect on human CRPC cells through the activation of the ER stress-autophagy axis and the rewiring of mitochondrial metabolism

Abundant Human DNA Contamination Identified in Non-Primate Genome Databases

During routine screens of the NCBI databases using human repetitive elements we discovered an unlikely level of nucleotide identity across a broad range of phyla. To ascertain whether databases containing DNA sequences, genome assemblies and trace archive reads were contaminated with human sequences, we performed an in depth search for sequences of human origin in non-human species. Using a primate specific SINE, AluY, we screened 2,749 non-primate public databases from NCBI, Ensembl, JGI, and UCSC and have found 492 to be contaminated with human sequence. These represent species ranging from bacteria (B. cereus) to plants (Z. mays) to fish (D. rerio) with examples found from most phyla. The identification of such extensive contamination of human sequence across databases and sequence types warrants caution among the sequencing community in future sequencing efforts, such as human re-sequencing. We discuss issues this may raise as well as present data that gives insight as to how this may be occurring

Non-Standard Neutrino Propagation and Pion Decay

Motivated by the findings of the OPERA experiment, we discuss the hypothesis that neutrino propagation does not obey Einstein special relativity. Under a minimal set of modifications of the standard model Lagrangian, we consider the implications of non standard neutrino propagation on the description of neutrino interactions and, specifically, on the pion decay processes. We show that all the different dispersion relations which have been proposed so far to explain OPERA results, imply huge departures from the standard expectations. The decay channel $\pi^+ \to e^+ \nu_{\rm e}$ becomes significantly larger than in the standard scenario, and may even dominate over $\pi^+ \to \mu^+ \nu_{\rm \mu}$. Moreover, the spectral distribution of neutrinos produced in the decay processes and the probability that a pion decays in flight in neutrinos show large deviations from the standard results.Comment: 17 pages, 10 figures, version accepted in JHE

Tor1/Sch9-Regulated Carbon Source Substitution Is as Effective as Calorie Restriction in Life Span Extension

The effect of calorie restriction (CR) on life span extension, demonstrated in organisms ranging from yeast to mice, may involve the down-regulation of pathways, including Tor, Akt, and Ras. Here, we present data suggesting that yeast Tor1 and Sch9 (a homolog of the mammalian kinases Akt and S6K) is a central component of a network that controls a common set of genes implicated in a metabolic switch from the TCA cycle and respiration to glycolysis and glycerol biosynthesis. During chronological survival, mutants lacking SCH9 depleted extracellular ethanol and reduced stored lipids, but synthesized and released glycerol. Deletion of the glycerol biosynthesis genes GPD1, GPD2, or RHR2, among the most up-regulated in long-lived sch9Δ, tor1Δ, and ras2Δ mutants, was sufficient to reverse chronological life span extension in sch9Δ mutants, suggesting that glycerol production, in addition to the regulation of stress resistance systems, optimizes life span extension. Glycerol, unlike glucose or ethanol, did not adversely affect the life span extension induced by calorie restriction or starvation, suggesting that carbon source substitution may represent an alternative to calorie restriction as a strategy to delay aging

Discovery of VHE Gamma Radiation from IC443 with the MAGIC Telescope

We report the detection of a new source of very high energy (VHE, E_gamma >= 100GeV) gamma-ray emission located close to the Galactic Plane, MAGIC J0616+225, which is spatially coincident with SNR IC443. The observations were carried out with the MAGIC telescope in the periods December 2005 - January 2006 and December 2006 - January 2007. Here we present results from this source, leading to a VHE gamma-ray signal with a statistical significance of 5.7 sigma in the 2006/7 data and a measured differential gamma-ray flux consistent with a power law, described as dN_gamma/(dA dt dE) = (1.0 +/- 0.2)*10^(-11)(E/0.4 TeV)^(-3.1 +/- 0.3) cm^(-2)s^(-1)TeV^(-1). We briefly discuss the observational technique used and the procedure implemented for the data analysis. The results are put in the perspective of the multiwavelength emission and the molecular environment found in the region of IC443.Comment: Accepted by ApJ Letter

Effects of bromopride on the healing of left colon anastomoses of rats

Objetivo: Avaliar os efeitos da bromoprida sobre a formação de aderências e a cicatrização de anastomoses de cólon esquerdo de ratos. Métodos: Foram incluídos 40 ratos, divididos em dois grupos contendo 20 animais, para administração de bromoprida (grupo de estudo- E) ou solução fisiológica (grupo controle- C). Cada grupo foi dividido em subgrupos contendo 10 animais cada, para eutanásia no terceiro (E3 e C3) ou no sétimo dia (E7 e C7) de pós-operatório. Os ratos foram submetidos à secção do cólon esquerdo e anastomose término-terminal. No dia da relaparotomia, foi avaliada a quantidade total de aderências e removido um segmento colônico contendo a anastomose para análise histopatológica, da força de ruptura e da concentração de hidroxiprolina. Resultados: Não houve diferença entre os grupos em relação à evolução clínica. Dois animais do grupo de estudo apresentaram deiscência de anastomose bloqueada. Os animais que receberam bromoprida apresentaram número de aderências intracavitárias e aderências à anastomose semelhantes ao grupo controle. As anastomoses dos animais do grupo E3 apresentaram menor resistência de ruptura do que as do grupo C3 (p=0,04). Este efeito não ocorreu no sétimo dia de pós-operatório (p=0,37). Não houve diferença significativa entre os grupos em relação à histopatologia ou concentração de hidroxiprolina das anastomoses. Conclusão: O uso da bromoprida está associado à diminuição da resistência tênsil de anastomoses do cólon esquerdo de ratos no terceiro dia de pós-operatório.Objective: To evaluate the effects of bromopride on the formation of adhesions and anastomotic healing in the left colon of rats. Methods: We divided 40 rats into two groups of 20 animals, administration of bromopride (study group-E) or saline (control group- C). Each group was divided into subgroups containing 10 animals each for euthanasia in the third (C3 and E3) or the seventh (E7 and C7) postoperative days. The rats were submitted to section of the left colon and end-to-end anastomosis. On the day of reoperation, we evaluated the total amount of adhesions and removed a colonic segment containing the anastomosis for histopathological analysis, assessment of rupture strength and hydroxyproline concentration. Results: There was no difference between groups in relation to clinical outcome. Two animals in the study group had blocked anastomotic leakage. The animals that received bromopride had the number of intracavitary adhesions and adhesions to the anastomosis similar to the control group. The anastomoses from the group E3 animals showed lower resistance to rupture the one from the C3 group (p = 0.04). This effect did not occur on the seventh postoperative day (p = 0.37). There was no significant difference between groups in relation to histopathology and hydroxyproline concentration in the anastomoses. Conclusion: The use of bromopride was associated with decreased tensile strength of left colon anastomosis in rats in the third postoperative day

Sperm Chromatin-Induced Ectopic Polar Body Extrusion in Mouse Eggs after ICSI and Delayed Egg Activation

Meiotic chromosomes in an oocyte are not only a maternal genome carrier but also provide a positional signal to induce cortical polarization and define asymmetric meiotic division of the oocyte, resulting in polar body extrusion and haploidization of the maternal genome. The meiotic chromosomes play dual function in determination of meiosis: 1) organizing a bipolar spindle formation and 2) inducing cortical polarization and assembly of a distinct cortical cytoskeleton structure in the overlying cortex for polar body extrusion. At fertilization, a sperm brings exogenous paternal chromatin into the egg, which induces ectopic cortical polarization at the sperm entry site and leads to a cone formation, known as fertilization cone. Here we show that the sperm chromatin-induced fertilization cone formation is an abortive polar body extrusion due to lack of spindle induction by the sperm chromatin during fertilization. If experimentally manipulating the fertilization process to allow sperm chromatin to induce both cortical polarization and spindle formation, the fertilization cone can be converted into polar body extrusion. This suggests that sperm chromatin is also able to induce polar body extrusion, like its maternal counterpart. The usually observed cone formation instead of ectopic polar body extrusion induced by sperm chromatin during fertilization is due to special sperm chromatin compaction which restrains it from rapid spindle induction and therefore provides a protective mechanism to prevent a possible paternal genome loss during ectopic polar body extrusion

Attainment of clinical performance targets and improvement in clinical outcomes and resource use in hemodialysis care: a prospective cohort study

BACKGROUND: Clinical performance targets are intended to improve patient outcomes in chronic disease through quality improvement, but evidence of an association between multiple target attainment and patient outcomes in routine clinical practice is often lacking. METHODS: In a national prospective cohort study (ESRD Quality, or EQUAL), we examined whether attainment of multiple targets in 668 incident hemodialysis patients from 74 U.S. not-for-profit dialysis clinics was associated with better outcomes. We measured whether the following accepted clinical performance targets were met at 6 months after study enrollment: albumin (≥4.0 g/dl), hemoglobin (≥11 g/dl), calcium-phosphate product (<55 mg(2)/dl(2)), dialysis dose (Kt/V≥1.2), and vascular access type (fistula). Outcomes included mortality, hospital admissions, hospital days, and hospital costs. RESULTS: Attainment of each of the five targets was associated individually with better outcomes; e.g., patients who attained the albumin target had decreased mortality [relative hazard (RH) = 0.55, 95% confidence interval (CI), 0.41–0.75], hospital admissions [incidence rate ratio (IRR) = 0.67, 95% CI, 0.62–0.73], hospital days (IRR = 0.61, 95% CI, 0.58–0.63), and hospital costs (average annual cost reduction = $3,282, P = 0.002), relative to those who did not. Increasing numbers of targets attained were also associated, in a graded fashion, with decreased mortality (P = 0.030), fewer hospital admissions and days (P < 0.001 for both), and lower costs (P = 0.029); these trends remained statistically significant for all outcomes after adjustment (P < 0.001), except cost, which was marginally significant (P = 0.052). CONCLUSION: Attainment of more clinical performance targets, regardless of which targets, was strongly associated with decreased mortality, hospital admissions, and resource use in hemodialysis patients