β-lapachone induces growth inhibition and apoptosis in bladder cancer cells by modulation of BCL-2 family and activation of caspases

Aim: To study in vitro the molecular mechanism of apoptosis caused by b-lapachone, a quinone obtained from the bark of the lapacho tree (Tabebuia avellanedae). Materials and Methods: The study was carried out on human bladder carcinoma T24 cell line. Determination of cell viability was done using trypan blue exclusion method, apoptosis quantitative estimation — by DAPI staining and agarose gel electrophoresis for DNA fragmentation. Flow cytometry analysis, RT-PCR and Western blot analysis, colorimetric assay of caspase activity were applied as well. Results: It was found that in micromolar range of concentrations b-lapachone inhibited the viability of T24 cells by inducing apoptosis, which could be proved by formation of apoptotic bodies and DNA fragmentation. Treatment of T24 cells with b-lapachone resulted in a down-regulation of Bcl-2 expression and up-regulation of Bax expression. b-lapachone-induced apoptosis was also associated with activation of caspase-3 and caspase-9, inhibition of IAP expression, and degradation of poly (ADP-ribose) polymerase, phospholipase C-g1 and b-catenin proteins. At the same time Fas and FasL levels were inhibited upon treatment with b-lapachone in a concentration-dependent manner. Conclusion: b-lapachone-induced apoptosis in T24 cells is mediated, at least in part, by the mitochondrial-signaling pathway.Цель: изучить механизмы апоптоза клеток карциномы мочевого пузыря человека Т24 при действии β-лапакона, хинона из коры дерева Tabebuia avellanedae. Материалы и методы: для определения жизнеспособности клеток использовали окраску трипановым синим; окрашивание DAPI и электрофоретический анализ фрагментации ДНК в агарозном геле, метод проточной цитометрии (для количественной оценки апоптоза); полимеразную цепную реакцию в режиме реального времени (РВ-ПЦР) и Вестерн блот-анализ (для оценки уровня экспрессии генов и белков), а также колориметрический анализ активности каспаз. Результаты: выявлено, что в микромолярных концентрациях β-лапакон понижает жизне- способность клеток линии Т24 путем активации апоптоза, что подтверждается формированием апоптотических тел и фрагментацией ДНК. Результаты РВ-ПЦР и иммуноблоттинга указывают на то, что обработка клеток β-лапаконом приводит к снижению экспрессии Bcl-2 и к активации Bax. Апоптоз, индуцированный β-лапаконом, также сопровож- дается активацией каспаз -3­­ и -9, ингибированием экспрессии семейства IAP, а также деградацией поли-(ADP-рибозо) полимеразы, фосфатазы C-γ1 и β-катенина. Тем не менее, уровень экспрессии Fas и FasL снижался при увеличении концентрации β-лапакона. Выводы: апоптоз, индуцированный при действии β-лапакона в клетках Т24, может быть час- тично опосредован митохондриальным сигнальным каскадом

Golgi Outpost Synthesis Impaired by Toxic Polyglutamine Proteins Contributes to Dendritic Pathology in Neurons

Dendrite aberration is a common feature of neurodegenerative diseases caused by protein toxicity, but the underlying mechanisms remain largely elusive. Here, we show that nuclear polyglutamine (polyQ) toxicity resulted in defective terminal dendrite elongation accompanied by a loss of Golgi outposts (GOPs) and a decreased supply of plasma membrane (PM) in Drosophila class IV dendritic arborization (da) (C4 da) neurons. mRNA sequencing revealed that genes downregulated by polyQ proteins included many secretory pathway-related genes, including COPII genes regulating GOP synthesis. Transcription factor enrichment analysis identified CREB3L1/CrebA, which regulates COPII gene expression. CrebA overexpression in C4 da neurons restores the dysregulation of COPII genes, GOP synthesis, and PM supply. Chromatin immunoprecipitation (ChIP)-PCR revealed that CrebA expression is regulated by CREB-binding protein (CBP), which is sequestered by polyQ proteins. Furthermore, co-overexpression of CrebA and Rac1 synergistically restores the polyQ-induced dendrite pathology. Collectively, our results suggest that GOPs impaired by polyQ proteins contribute to dendrite pathology through the CBP-CrebA-COPII pathway. ? 2017 The Author(s)113Ysciescopu

Dalitz plot analysis of D_s+ and D+ decay to pi+pi-pi+ using the K-matrix formalism

FOCUS results from Dalitz plot analysis of D_s+ and D+ to pi+pi-pi+ are presented. The K-matrix formalism is applied to charm decays for the first time to fully exploit the already existing knowledge coming from the light-meson spectroscopy experiments. In particular all the measured dynamics of the S-wave pipi scattering, characterized by broad/overlapping resonances and large non-resonant background, can be properly included. This paper studies the extent to which the K-matrix approach is able to reproduce the observed Dalitz plot and thus help us to understand the underlying dynamics. The results are discussed, along with their possible implications on the controversial nature of the sigma meson.Comment: To be submitted to Phys.Lett.B A misprint corrected in formula

Search for Λc+→pK+π−\Lambda_c^+ \to p K^+ \pi^- and Ds+→K+K+π−D_s^+ \to K^+ K^+ \pi^- Using Genetic Programming Event Selection

We apply a genetic programming technique to search for the double Cabibbo suppressed decays $\Lambda_c^+ \to p K^+ \pi^-$ and $D_s^+ \to K^+ K^+ \pi^-$. We normalize these decays to their Cabibbo favored partners and find $\text{BR}($\Lambda_c^+ \to p K^+ \pi^-$)/\text{BR}($\Lambda_c^+ \to p K^- \pi^+$) = (0.05 \pm 0.26 \pm 0.02)$ and $\text{BR}($D_s^+ \to K^+ K^+ \pi^-$)/\text{BR}($D_s^+ \to K^+ K^- \pi^+$) = (0.52\pm 0.17\pm 0.11)$ where the first errors are statistical and the second are systematic. Expressed as 90% confidence levels (CL), we find $< 0.46$ and $< 0.78$ respectively. This is the first successful use of genetic programming in a high energy physics data analysis.Comment: 10 page

Measurement of the D+ and Ds+ decays into K+K-K+

We present the first clear observation of the doubly Cabibbo suppressed decay D+ --> K-K+K+ and the first observation of the singly Cabibbo suppressed decay Ds+ --> K-K+K+. These signals have been obtained by analyzing the high statistics sample of photoproduced charm particles of the FOCUS(E831) experiment at Fermilab. We measure the following relative branching ratios: Gamma(D+ --> K-K+K+)/Gamma(D+ --> K-pi+pi+) = (9.49 +/- 2.17(statistical) +/- 0.22(systematic))x10^-4 and Gamma(Ds+ --> K-K+K+)/Gamma(Ds+ --> K-K+pi+) = (8.95 +/- 2.12(statistical) +2.24(syst.) -2.31(syst.))x10^-3.Comment: 10 pages, 8 figure

A Non-parametric Approach to the D+ to K*0bar mu+ nu Form Factors

Using a large sample of D+ -> K- pi+ mu+ nu decays collected by the FOCUS photoproduction experiment at Fermilab, we present the first measurements of the helicity basis form factors free from the assumption of spectroscopic pole dominance. We also present the first information on the form factor that controls the s-wave interference discussed in a previous paper by the FOCUS collaboration. We find reasonable agreement with the usual assumption of spectroscopic pole dominance and measured form factor ratios.Comment: 14 pages, 5 figures, and 2 tables. We updated the previous version by changing some words, removing one plot, and adding two tables. These changes are mostly stylisti

Measurements of Ξc+\Xi_c^{+} Branching Ratios

Using data collected by the fixed target Fermilab experiment FOCUS, we measure the branching ratios of the Cabibbo favored decays $\Xi_c^+ \to \Sigma^+K^-\pi^+$, $\Xi_c^+ \to \Sigma^+ \bar{K}^{*}(892)^0$, and $\Xi_c^+ \to \Lambda^0K^-\pi^+\pi^+$ relative to $\Xi_c^+ \to \Xi^-\pi^+\pi^+$ to be $0.91\pm0.11\pm0.04$, $0.78\pm0.16\pm0.06$, and $0.28\pm0.06\pm0.06$, respectively. We report the first observation of the Cabibbo suppressed decay $\Xi_c^+ \to \Sigma^+K^+K^-$ and we measure the branching ratio relative to $\Xi_c^+ \to \Sigma^+K^-\pi^+$ to be $0.16\pm0.06\pm0.01$. We also set 90% confidence level upper limits for $\Xi_c^+ \to \Sigma^+ \phi$ and $\Xi_c^+ \to \Xi^*(1690)^0(\Sigma^+ K^-) K^+$ relative to $\Xi_c^+ \to \Sigma^+K^-\pi^+$ to be 0.12 and 0.05, respectively. We find an indication of the decays $\Xi_c^+ \to \Omega^-K^{+}\pi^+$ and $\Xi_c^+ \to \Sigma^{*}(1385)^+ \bar{K}^0$ and set 90% confidence level upper limits for the branching ratios with respect to $\Xi_c^+ \to \Xi^-\pi^+\pi^+$ to be 0.12 and 1.72, respectively. Finally, we determine the 90% C.L. upper limit for the resonant contribution $\Xi_c^+ \to \Xi^{*}(1530)^0 \pi^+$ relative to $\Xi_c^+ \to \Xi^-\pi^+\pi^+$ to be 0.10.Comment: 14 pages, 8 figure

Measurement of the branching ratio of the decay D^0 -> \pi^-\mu^+\nu relative to D^0 -> K^-\mu^+\nu

We present a new measurement of the branching ratio of the Cabibbo suppressed decay D^0\to \pi^-\mu^+\nu relative to the Cabibbo favored decay D^0\to K^-\mu^+\nu and an improved measurement of the ratio |\frac{f_+^{\pi}(0)}{f_+^{K}(0)}|. Our results are 0.074 \pm 0.008 \pm 0.007 for the branching ratio and 0.85 \pm 0.04 \pm 0.04 \pm 0.01 for the form factor ratio, respectively.Comment: 13pages, 3 figure