CHK1 expression in gastric cancer is modulated by p53 and RB1/E2F1: Implications in chemo/radiotherapy response

Radiation has a limited but relevant role in the adjuvant therapy of gastric cancer (GC) patients. Since Chk1 plays a critical function in cellular response to genotoxic agents, we aimed to analyze the role of Chk1 in GC as a biomarker for radiotherapy resistance. We analyzed Chk1 expression in AGS and MKN45 human GC cell lines by RT-QPCR and WB and in a small cohort of human patient’s samples. We demonstrated that Chk1 overexpression specifically increases resistance to radiation in GC cells. Accordingly, abrogation of Chk1 activity with UCN-01 and its expression with shChk1 increased sensitivity to bleomycin and radiation. Furthermore, when we assessed Chk1 expression in human samples, we found a correlation between nuclear Chk1 accumulation and a decrease in progression free survival. Moreover, using a luciferase assay we found that Chk1’s expression is controlled by p53 and RB/E2F1 at the transcriptional level. Additionally, we present preliminary data suggesting a posttranscriptional regulation mechanism, involving miR-195 and miR-503, which are inversely correlated with expression of Chk1 in radioresistant cells. In conclusion, Chk1/microRNA axis is involved in resistance to radiation in GC, and suggests Chk1 as a potential tool for optimal stratification of patients susceptible to receive adjuvant radiotherapy after surgeryThis work was supported by Instituto de Salud Carlos III–Fondo de Investigación Sanitaria (PS09/1988 to ISP; PI11-00949, pI014-1495 and Feder Funds to RP); Comunidad Autónoma de Madrid-Universidad Autónoma de Madrid (CCG10-UAM/BIO-5871 to ISP); Fundación Leticia Castillejo Castillo and Ministerio de Ciencia e Innovación (SAF2012-30862 to RSP), Spain

Paper-based chromatic toxicity bioassay by analysis of bacterial ferricyanide reduction

Water quality assessment requires a continuous and strict analysis of samples to guarantee compliance with established standards. Nowadays, the increasing number of pollutants and their synergistic effects lead to the development general toxicity bioassays capable to analyse water pollution as a whole. Current general toxicity methods, e.g. Microtox®, rely on long operation protocols, the use of complex and expensive instrumentation and sample pre-treatment, which should be transported to the laboratory for analysis. These requirements delay sample analysis and hence, the response to avoid an environmental catastrophe. In an attempt to solve it, a fast (15 min) and low-cost toxicity bioassay based on the chromatic changes associated to bacterial ferricyanide reduction is here presented. E. coli cells (used as model bacteria) were stably trapped on low-cost paper matrices (cellulose-based paper discs, PDs) and remained viable for long times (1 month at -20 °C). Apart from bacterial carrier, paper matrices also acted as a fluidic element, allowing fluid management without the need of external pumps. Bioassay evaluation was performed using copper as model toxic agent. Chromatic changes associated to bacterial ferricyanide reduction were determined by three different transduction methods, i.e. (i) optical reflectometry (as reference method), (ii) image analysis and (iii) visual inspection. In all cases, bioassay results (in terms of half maximal effective concentrations, EC50) were in agreement with already reported data, confirming the good performance of the bioassay. The validation of the bioassay was performed by analysis of real samples from natural sources, which were analysed and compared with a reference method (i.e. Microtox). Obtained results showed agreement for about 70% of toxic samples and 80% of non-toxic samples, which may validate the use of this simple and quick protocol in the determination of general toxicity. The minimum instrumentation requirements and the simplicity of the bioassay open the possibility of in-situ water toxicity assessment with a fast and low-cost protocolPostprint (author's final draft

Continuously-variable survival exponent for random walks with movable partial reflectors

We study a one-dimensional lattice random walk with an absorbing boundary at the origin and a movable partial reflector. On encountering the reflector, at site x, the walker is reflected (with probability r) to x-1 and the reflector is simultaneously pushed to x+1. Iteration of the transition matrix, and asymptotic analysis of the probability generating function show that the critical exponent delta governing the survival probability varies continuously between 1/2 and 1 as r varies between 0 and 1. Our study suggests a mechanism for nonuniversal kinetic critical behavior, observed in models with an infinite number of absorbing configurations.Comment: 5 pages, 3 figure

Mapping of the human visual cortex using image-guided transcranial magnetic stimulation

We describe a protocol using transcranial magnetic stimulation (TMS) to systematically map the visual sensations induced by focal and non-invasive stimulation of the human occipital cortex. TMS is applied with a figure of eight coil to 28 positions arranged in a 232-cm grid over the occipital area. A digitizing tablet connected to a PC computer running customized software, and audio and video recording are used for detailed and accurate data collection and analysis of evoked phosphenes. A frameless image-guided neuronavigational device is used to describe the position of the actual sites of the stimulation coils relative to the cortical surface. Our results show that TMS is able to elicit phosphenes in almost all sighted subjects and in a proportion of blind subjects. Evoked phosphenes are topographically organized. Despite minor inter-individual variations, the mapping results are reproducible and show good congruence among different subjects. This procedure has potential to improve our understanding of physiologic organization and plastic changes in the human visual system and to establish the degree of remaining functional visual cortex in blind subjects. Such a non-invasive method is critical for selection of suitable subjects for a cortical visual prosthesis.This research has been carried out with financial support from the Commission of the European Communities, specific RTD programme ‘Quality of Life and Management of Living Resources’, QLK6-CT-2001-00279 and by the Ministerio de Ciencia y Tecnologıa(MAT2000-1049)Fondo de Investigaciones de la Seguridad Social (FISS 01-0674)National Institute of Mental Health (MH60734, MH57980)National Eye Institute (EYEY12091)Harvard-Thorndike General Clinical Research Center at Beth Israel Deaconess Medical Center (NCRR MO1 RR01032).Medicin

An in-depth view of the microscopic dynamics of Ising spin glasses at fixed temperature

Using the dedicated computer Janus, we follow the nonequilibrium dynamics of the Ising spin glass in three dimensions for eleven orders of magnitude. The use of integral estimators for the coherence and correlation lengths allows us to study dynamic heterogeneities and the presence of a replicon mode and to obtain safe bounds on the Edwards-Anderson order parameter below the critical temperature. We obtain good agreement with experimental determinations of the temperature-dependent decay exponents for the thermoremanent magnetization. This magnitude is observed to scale with the much harder to measure coherence length, a potentially useful result for experimentalists. The exponents for energy relaxation display a linear dependence on temperature and reasonable extrapolations to the critical point. We conclude examining the time growth of the coherence length, with a comparison of critical and activated dynamics.Comment: 38 pages, 26 figure

Matching microscopic and macroscopic responses in glasses

We first reproduce on the Janus and Janus II computers a milestone experiment that measures the spin-glass coherence length through the lowering of free-energy barriers induced by the Zeeman effect. Secondly we determine the scaling behavior that allows a quantitative analysis of a new experiment reported in the companion Letter [S. Guchhait and R. Orbach, Phys. Rev. Lett. 118, 157203 (2017)]. The value of the coherence length estimated through the analysis of microscopic correlation functions turns out to be quantitatively consistent with its measurement through macroscopic response functions. Further, non-linear susceptibilities, recently measured in glass-forming liquids, scale as powers of the same microscopic length.Comment: 6 pages, 4 figure

Uso do cobre no controle da verminose gastrintestinal em ovinos.

No RS, a prevalência de resistência anti-helmíntica nos rebanhos ovinos representa um grave problema para o controle eficiente das helmintoses gastrintestinais. A resistência anti-helmíntica em nematódeos de ovinos se constitui no principal problema sanitário com que se defronta a indústria ovinícola no Brasil'. Esta situação não é diferente em outros países onde a resistência dos parasitos à maioria dos grupos químicos é uma realidade. Devido a estes fatores, vários estudos vem sendo realizados visando o desenvolvimento de métodos alternativos que contribuam para reduzir o consumo do uso de anti-helmínticos no controle da gastroenterite verminótica, como o uso de fungos nematófagose o emprego de minerais, cobre, como exemplo. Assim, pesquisadores da Nova Zelândia, relataram que a administração oral de partículas de óxido de cobre (COWP) para ovinos proporcionava uma redução significativa no número de parasitos recuperados na necropsia, com uma redução de 96% para Haemonchus. contortus.e 56% para Ostertagia circumcincta, respectivamente, além disso, aumento da mortalidade e redução da fecundidade de fêmeas de H. contortus também tem sido relatado, bem como, beneficios no combate de parasitos abomasals '. Este estudo teve o objetivo de verificar o tempo de proteção do cobre contra reinfecções por helmintos gastrintestinais de ovinos em pastejo para possível incorporação como ferramenta alternativa de controle