Mifepristone increases mRNA translation rate, triggers the unfolded protein response, increases autophagic flux, and kills ovarian cancer cells in combination with proteasome or lysosome inhibitors

The synthetic steroid mifepristone blocks the growth of ovarian cancer cells, yet the mechanism driving such effect is not entirely understood. Unbiased genomic and proteomic screenings using ovarian cancer cell lines of different genetic backgrounds and sensitivities to platinum led to the identification of two key genes upregulated by mifepristone and involved in the unfolded protein response (UPR): the master chaperone of the endoplasmic reticulum (ER), glucose regulated protein (GRP) of 78 kDa, and the CCAAT/enhancer binding protein homologous transcription factor (CHOP). GRP78 and CHOP were upregulated by mifepristone in ovarian cancer cells regardless of p53 status and platinum sensitivity. Further studies revealed that the three UPR-associated pathways, PERK, IRE1α, and ATF6, were activated by mifepristone. Also, the synthetic steroid acutely increased mRNA translation rate, which, if prevented, abrogated the splicing of XBP1 mRNA, a non-translatable readout of IRE1α activation. Moreover, mifepristone increased LC3-II levels due to increased autophagic flux. When the autophagic–lysosomal pathway was inhibited with chloroquine, mifepristone was lethal to the cells. Lastly, doses of proteasome inhibitors that are inadequate to block the activity of the proteasomes, caused cell death when combined with mifepristone; this phenotype was accompanied by accumulation of poly-ubiquitinated proteins denoting proteasome inhibition. The stimulation by mifepristone of ER stress and autophagic flux offers a therapeutic opportunity for utilizing this compound to sensitize ovarian cancer cells to proteasome or lysosome inhibitors.Fil: Zhang, Lei. University Of South Dakota; Estados UnidosFil: Hapon, María Belén. University Of South Dakota; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Goyeneche, Alicia A.. University Of South Dakota; Estados Unidos. McGill University; CanadáFil: Srinivasan, Rekha. University Of South Dakota; Estados UnidosFil: Gamarra Luques, Carlos Diego. University Of South Dakota; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Callegari, Eduardo A.. University Of South Dakota; Estados UnidosFil: Drappeau, Donis D.. University Of South Dakota; Estados UnidosFil: Terpstra, Erin J.. University Of South Dakota; Estados UnidosFil: Pan, Bo. University Of South Dakota; Estados UnidosFil: Knapp, Jennifer R.. University of Kansas; Estados UnidosFil: Chien, Jeremy. University of Kansas; Estados UnidosFil: Wang, Xuejun. University Of South Dakota; Estados UnidosFil: Eyster, Kathleen M.. University Of South Dakota; Estados UnidosFil: Telleria, Carlos Marcelo. University Of South Dakota; Estados Unidos. McGill University; Canadá. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Can luteal regression be reversed?

The corpus luteum is an endocrine gland whose limited lifespan is hormonally programmed. This debate article summarizes findings of our research group that challenge the principle that the end of function of the corpus luteum or luteal regression, once triggered, cannot be reversed. Overturning luteal regression by pharmacological manipulations may be of critical significance in designing strategies to improve fertility efficacy

Coarse-grained entanglement classification through orthogonal arrays

Classification of entanglement in multipartite quantum systems is an open problem solved so far only for bipartite systems and for systems composed of three and four qubits. We propose here a coarse-grained classification of entanglement in systems consisting of $N$ subsystems with an arbitrary number of internal levels each, based on properties of orthogonal arrays with $N$ columns. In particular, we investigate in detail a subset of highly entangled pure states which contains all states defining maximum distance separable codes. To illustrate the methods presented, we analyze systems of four and five qubits, as well as heterogeneous tripartite systems consisting of two qubits and one qutrit or one qubit and two qutrits.Comment: 38 pages, 1 figur

Mifepristone prevents repopulation of ovarian cancer cells escaping cisplatin-paclitaxel therapy

<p>Abstract</p> <p>Background</p> <p>Advanced ovarian cancer is treated with cytoreductive surgery and combination platinum- and taxane-based chemotherapy. Although most patients have acute clinical response to this strategy, the disease ultimately recurs. In this work we questioned whether the synthetic steroid mifepristone, which as monotherapy inhibits the growth of ovarian cancer cells, is capable of preventing repopulation of ovarian cancer cells if given after a round of lethal cisplatin-paclitaxel combination treatment.</p> <p>Methods</p> <p>We established an <it>in vitro</it> approach wherein ovarian cancer cells with various sensitivities to cisplatin or paclitaxel were exposed to a round of lethal doses of cisplatin for 1 h plus paclitaxel for 3 h. Thereafter, cells were maintained in media with or without mifepristone, and short- and long-term cytotoxicity was assessed.</p> <p>Results</p> <p>Four days after treatment the lethality of cisplatin-paclitaxel was evidenced by reduced number of cells, increased hypodiploid DNA content, morphological features of apoptosis, DNA fragmentation, and cleavage of caspase-3, and of its downstream substrate PARP. Short-term presence of mifepristone either enhanced or did not modify such acute lethality. Seven days after receiving cisplatin-paclitaxel, cultures showed signs of relapse with escaping colonies that repopulated the plate in a time-dependent manner. Conversely, cultures exposed to cisplatin-paclitaxel followed by mifepristone not only did not display signs of repopulation following initial chemotherapy, but they also had their clonogenic capacity drastically reduced when compared to cells repopulating after cisplatin-paclitaxel.</p> <p>Conclusions</p> <p>Cytostatic concentrations of mifepristone after exposure to lethal doses of cisplatin and paclitaxel in combination blocks repopulation of remnant cells surviving and escaping the cytotoxic drugs.</p

The celiac ganglion modulates LH-induced inhibition of androstenedione release in late pregnant rat ovaries

BACKGROUND: Although the control of ovarian production of steroid hormones is mainly of endocrine nature, there is increasing evidence that the nervous system also influences ovarian steroidogenic output. The purpose of this work was to study whether the celiac ganglion modulates, via the superior ovarian nerve, the anti-steroidogenic effect of LH in the rat ovary. Using mid- and late-pregnant rats, we set up to study: 1) the influence of the noradrenergic stimulation of the celiac ganglion on the ovarian production of the luteotropic hormone androstenedione; 2) the modulatory effect of noradrenaline at the celiac ganglion on the anti-steroidogenic effect of LH in the ovary; and 3) the involvement of catecholaminergic neurotransmitters released in the ovary upon the combination of noradrenergic stimulation of the celiac ganglion and LH treatment of the ovary. METHODS: The ex vivo celiac ganglion-superior ovarian nerve-ovary integrated system was used. This model allows studying in vitro how direct neural connections from the celiac ganglion regulate ovarian steroidogenic output. The system was incubated in buffer solution with the ganglion and the ovary located in different compartments and linked by the superior ovarian nerve. Three experiments were designed with the addition of: 1) noradrenaline in the ganglion compartment; 2) LH in the ovarian compartment; and 3) noradrenaline and LH in the ganglion and ovarian compartments, respectively. Rats of 15, 19, 20 and 21 days of pregnancy were used, and, as an end point, the concentration of the luteotropic hormone androstenedione was measured in the ovarian compartment by RIA at various times of incubation. For some of the experimental paradigms the concentration of various catecholamines (dihydroxyphenylalanine, dopamine, noradrenaline and adrenaline) was also measured in the ovarian compartment by HPLC. RESULTS: The most relevant result concerning the action of noradrenaline in the celiac ganglion was found on day 21 of pregnancy resulting in the inhibition of androstenedione release from the ovarian compartment. In addition on day 15 of pregnancy, LH placed in the ovarian compartment led to an inhibition of the release of androstenedione, and this inhibitory effect was further reinforced by the joint action of noradrenaline in the celiac ganglion and LH in the ovary. The levels of catecholamines in the ovarian compartment showed differences among the experiments; of significance, the joint treatment of noradrenaline in the celiac ganglion and LH in the ovary resulted in a remarkable increase in the ovarian levels of noradrenaline and adrenaline when compared to the effect achieved by either one of the compounds added alone. CONCLUSION: Our results demonstrate that the noradrenergic stimulation of the celiac ganglion reinforces the LH-induced inhibition of androstenedione production by the ovary of late pregnant rats, and that this effect is associated with marked changes in the release of catecholamines in the ovary

Proteína cruda en Paspalum notatum, según diferentes niveles de fertilización nitrogenada [Resumen de Poster].

El campo natural es un ecosistema biodiverso que a través de su producción de forraje ofrece la principal fuente nutricional para la ganadería uruguaya. Debido a esto, disponer de especies nativas caracterizadas por su adaptación a diferentes ambientes, es fundamental para diseñar estrategias de reintroducción en ambientes degradados por sobrepastoreo o expansión del área agrícola. La especie Paspalum notatum (pasto horqueta) es una de las principales gramíneas perennes estivales presentes en las pasturas naturales de Uruguay. En el marco del desarrollo de una línea de investigación tendiente a mejorar los recursos genéticos de P. notatum en el Uruguay se realizó en 2006 una colecta en 97 ubicaciones representativas de todo el país, resultando en la caracterización genética y fenotípica, siendo la variedad INIA Sepé resultado de este trabajo de mejoramiento desarrollado en INIA Tacuarembó, Uruguay. En este marco el objetivo principal de este trabajo es la evaluación de la calidad nutricional de esta variedad en función de la variación del nivel de fertilización nitrogenada. Tradicionalmente, los parámetros de calidad de pasturas son aquellos que corresponden a su composición química: contenido de proteína cruda, materia grasa, cenizas, fibra detergente ácido y neutro (FDA y FDN) y la digestibilidad de la materia orgánica. De estos parámetros de calidad, los importantes desde el punto de la producción animal son el contenido de proteína cruda y la digestibilidad. El experimento fue sembrado en suelo de brunosoles éutricos en la estación experimental de INIA La Estanzuela, se utilizó un diseño experimental de bloques completos al azar con tres repeticiones donde se evaluaron cuatro niveles de nitrógeno: 0, 100, 300 y 500 kg de N.ha-1 , con fertilización en 4 etapas de 25 %. Durante su ciclo de desarrollo se efectuaron 4 cortes, resultando en 12 muestras por duplicado para evaluación de calidad. El contenido de nitrógeno se evaluó mediante el método de referencia Kjeldahl según A.O.A.C. 2001.11. Este tipo de ensayos resulta laborioso para el trabajo con un alto número de muestras. Por lo tanto, un segundo objetivo de este trabajo es estimar el desempeño de dos métodos secundarios que se encuentran a disposición en los laboratorios del Campus INIA- UdelaR, especialmente un equipo de espectroscopía de infrarrojo cercano (NIR, DA 7250, Perten) y un analizador elemental (CHN 628, LECO). Los resultados muestran que el óptimo de materia seca se corresponde a una cantidad de N de entre 300-500 kg de N.ha-1 . De los resultados de correlación de los métodos secundarios, el análisis de proteína cruda permitió obtener coeficientes de regresión lineal de 0.96 y 0.93, y una diferencia en el error de estimación del 0.7% y 1% en el caso del análisis por analizador elementar y NIR, respectivamente. Estos resultados primarios apuntan a la validación del análisis por analizador elemental en este tipo de muestras. Asimismo, el método de NIR es un buen método de screening, cuya calibración puede ser ajustada para responder específicamente a esta variedad