Efecto de la alimentación con pienso húmedo durante la lactación sobre los rendimientos productivos en cerdas

[EN] This work aims to compare different production parameters in two groups of sows during lactation fed wet or dry feed. 400 breeding females fed with the same commercial feed and housed in two types of maternity w[ES] El presente trabajo tiene por objeto comparar diversos parámetros productivos en dos grupos de cerdas alimentadas durante la lactación con pienso húmedo o seco. Se utilizaron 400 hembras reproductoras alimentaAlcácer Montañana, FJ. (2014). Efecto de la alimentación con pienso húmedo durante la lactación sobre los rendimientos productivos en cerdas. http://hdl.handle.net/10251/47439Archivo delegad

Glucocorticoid receptor antagonism overcomes resistance to BRAF inhibition in BRAFV600E-mutated metastatic melanoma

Clinical applications of glucocorticoids (GC) in Oncology are dependent on their pro-apoptotic action to treat lymphoproliferative cancers, and to alleviate side effects induced by chemotherapy and/or radiotherapy. However, the mechanism(s) by which GC may also promote tumor progression remains unclear. GC receptor (GR) knockdown decreases the antioxidant protection of highly metastatic B16-F10 melanoma cells. We hypothesize that a GR antagonist (RU486, mifepristone) could increase the efficacy of BRAF-related therapy in BRAFV600E-mutated metastatic melanoma. In vivo formed spontaneous skin tumors were reinoculated into nude mice to expand the metastases of different human BRAFV600E melanoma cells. The GR content of melanoma cell lines was measured by [3H]-labeled ligand binding assay. Nuclear Nrf2 and its transcription activity was investigated by RT-PCR, western blotting, and by measuring Nrf2- and redox state-related enzyme activities and metabolites. GR knockdown was achieved using lentivirus, and GR overexpression by transfection with the NR3C1 plasmid. shRNA-induced selective Bcl-xL, Mcl-1, AKT1 or NF-κB/p65 depletion was used to test the efficacy of vemurafenib (VMF) and RU486 against BRAFV600E-mutated metastatic melanoma. During early progression of skin melanoma metastases, RU486 and VMF induced a drastic metastases regression. However, treatment at an advanced stage of growth demonstrated the development of resistance to RU486 and VMF. This resistance was mechanistically linked to overexpression of specific proteins of the Bcl-2 family (Bcl-xL and Mcl-1 in our experimental models). We found that melanoma resistance is decreased if AKT and NF-κB signaling pathways are blocked. Our results highlight mechanisms by which metastatic melanoma cells adapt to survive.Medicin

Glucocorticoid receptor antagonism overcomes resistance to BRAF inhibition in BRAFV600E-mutated metastatic melanoma

Clinical applications of glucocorticoids (GC) in Oncology are dependent on their pro-apoptotic action to treat lymphoproliferative cancers, and to alleviate side effects induced by chemotherapy and/or radiotherapy. However, the mechanism(s) by which GC may also promote tumor progression remains unclear. GC receptor (GR) knockdown decreases the antioxidant protection of highly metastatic B16-F10 melanoma cells. We hypothesize that a GR antagonist (RU486, mifepristone) could increase the efficacy of BRAF-related therapy in BRAFV600E-mutated metastatic melanoma. In vivo formed spontaneous skin tumors were reinoculated into nude mice to expand the metastases of different human BRAFV600E melanoma cells. The GR content of melanoma cell lines was measured by [3H]-labeled ligand binding assay. Nuclear Nrf2 and its transcription activity was investigated by RT-PCR, western blotting, and by measuring Nrf2- and redox state-related enzyme activities and metabolites. GR knockdown was achieved using lentivirus, and GR overexpression by transfection with the NR3C1 plasmid. shRNA-induced selective Bcl-xL, Mcl-1, AKT1 or NF-κB/p65 depletion was used to test the efficacy of vemurafenib (VMF) and RU486 against BRAFV600E-mutated metastatic melanoma. During early progression of skin melanoma metastases, RU486 and VMF induced a drastic metastases regression. However, treatment at an advanced stage of growth demonstrated the development of resistance to RU486 and VMF. This resistance was mechanistically linked to overexpression of specific proteins of the Bcl-2 family (Bcl-xL and Mcl-1 in our experimental models). We found that melanoma resistance is decreased if AKT and NF-κB signaling pathways are blocked. Our results highlight mechanisms by which metastatic melanoma cells adapt to survive

Dictamen sobre la constitucionalidad de la pena de prisión permanente revisable

En este Dictamen se examina si resulta conforme a la Constitución española la pena de prisión permanente revisable introducida en el Código penal español por la LO 1/2015 (en concreto, en los artículos 33.2.a, 35, 36, 76.1.e, 78 bis, 92, 140, 485.1, 605.1, 607.1.1º, 607.1.2º y 607 bis 2.1º, todos ellos en la redacción que les dio la LO 1/2015). Y se concluye que no es así, porque la regulación de dicha pena vulnera la prohibición de penas inhumanas (art. 15.1 CE), el derecho a la libertad, porque prevé una privación de la misma desproporcionada y ajena a criterios de culpabilidad (art. 17.1 CE), el mandato de determinación derivado del principio de legalidad penal (art. 25.1 CE), y el mandato de resocialización (art. 25.2 CE), por cuanto prácticamente restringe toda posibilidad de resocialización del condenado

Selective targeting of collagen IV in the cancer cell microenvironment reduces tumor burden

Goodpasture antigen-binding protein (GPBP) is an exportable1 Ser/Thr kinase that induces collagen IV expansion and has been associated with chemoresistance following epithelial-to-mesenchymal transition (EMT). Here we demonstrate that cancer EMT phenotypes secrete GPBP (mesenchymal GPBP) which displays a predominant multimeric oligomerization and directs the formation of previously unrecognized mesh collagen IV networks (mesenchymal collagen IV). Yeast twohybrid (YTH) system was used to identify a 260SHCIE264 motif critical for multimeric GPBP assembly which then facilitated design of a series of potential peptidomimetics. The compound 3-[4''-methoxy-3,2'-dimethyl-(1,1';4',1'')terphenyl-2''-yl]propionic acid, or T12, specifically targets mesenchymal GPBP and disturbs its multimerization without affecting kinase catalytic site. Importantly, T12 reduces growth and metastases of tumors populated by EMT phenotypes. Moreover, low-dose doxorubicin sensitizes epithelial cancer precursor cells to T12, thereby further reducing tumor load. Given that T12 targets the pathogenic mesenchymal GPBP, it does not bind significantly to normal tissues and therapeutic dosing was not associated with toxicity. T12 is a first-in-class drug candidate to treat cancer by selectively targeting the collagen IV of the tumor cell microenvironment.This work was supported by grants: PET 2006_0721, TRA2009_0026, IPT-010000-2010-45, IPT-2011-1527- 010000, RTC-2014-2415-1, PCB-010000-2010-031, PCB- 010000-2010-032, EQU-2014-1-0301 of the Plan Nacional de Investigación, Desarrollo e Innovación of the Spanish Government and IMGESA/06/78, IMGESA/06/79 of Conselleria d’Empresa, Universitat i Ciencia of Generalitat Valenciana to Fibrostatin, S.L. and J.S.; SAF 2001/0453, SAF 2003-09772-C03-01, SAF 2006-12520-C02-01, SAF 2009-10703 of the Plan Nacional de Investigación, Desarrollo e Innovación of the Spanish Government and PROMETEO/2009/065, PROMETEOII/2014/048 of Conselleria de Educaciò of Generalitat Valenciana to J.S. Additional funding came from ERESA, BioStratum Inc. and NephroGenex Inc. R&D programs, and personal funding from Vicente Saus and Carmen Cano to J.S.. Torres Quevedo program of the Spanish Government granted F.R., F-R-R., R.B., E.L-P and A.P-S.Medicin

Glucocorticoid receptor knockdown decreases the antioxidant protection of B16 melanoma cells: an endocrine system-related mechanism that compromises metastatic cell resistance to vascular endothelium-induced tumor cytotoxicity.

We previously reported an interorgan system in which stress-related hormones (corticosterone and noradrenaline), interleukin-6, and glutathione (GSH) coordinately regulate metastatic growth of highly aggressive B16-F10 melanoma cells. Corticosterone, at levels measured in tumor-bearing mice, also induces apoptotic cell death in metastatic cells with low GSH content. In the present study we explored the potential role of glucocorticoids in the regulation of metastatic cell death/survival during the early stages of organ invasion. Glucocorticoid receptor (GCR) knockdown decreased the expression and activity of γ-glutamylcysteine synthetase (γ-GCS), the rate-limiting step in GSH synthesis, in metastatic cells in vivo independent of the tumor location (liver, lung, or subcutaneous). The decrease in γ-GCS activity was associated with lower intracellular GSH levels. Nrf2- and p53-dependent down-regulation of γ-GCS was associated with a decrease in the activities of superoxide dismutase 1 and 2, catalase, glutathione peroxidase, and glutathione reductase, but not of the O2--generating NADPH oxidase. The GCR knockdown-induced decrease in antioxidant protection caused a drastic decrease in the survival of metastatic cells during their interaction with endothelial cells, both in vitro and in vivo; only 10% of cancer cells attached to the endothelium survived compared to 90% survival observed in the controls. This very low rate of metastatic cell survival was partially increased (up to 52%) in vivo by inoculating B16-F10 cells preloaded with GSH ester, which enters the cell and delivers free GSH. Taken together, our results indicate that glucocorticoid signaling influences the survival of metastatic cells during their interaction with the vascular endothelium

of glucocorticoid receptor knockdown on the rates of GSH synthesis and efflux in iB16 melanoma cells.

<p>(A–C) Melanoma cells were isolated from the liver or lungs 7 days after inoculation and from subcutaneous tumors 14 days after inoculation for culture. Glutathione efflux corresponded to GSH because GSSG was, in all conditions, 1–2% of the total glutathione found in the extracellular space (not shown). To prevent degradation of the GSH accumulated in the extracellular space, γ-GT was blocked by adding 10 µM acivicin to the culture medium 2 h before measuring efflux. Enzyme activities were measured 22 h after seeding. Results obtained in iB16 cells transfected with lentiviral vector not harboring any gene (negative control) were not different from control values (not shown). Data are mean values ± S.D. (n = 9–10 in all cases). *p<0.05,**p<0.01<i>versus</i> iB16 controls. +p<0.05, ++p<0.01 <i>versus</i> melanoma cells isolated from liver metastases. (D) γ-GCS-HS and γ-GCS-LS expression was determined in cells cultured for 24h (previously isolated from <i>in vivo</i> tumors). Data, expressed as a fold change, show mean values ± S.D. from 5 to 6 different experiments. *p<0.01 <i>versus</i> iB16 cells.</p

Glucocorticoid receptor knockdown and GSH content in B16 melanoma cell subsets; and plasma corticosterone, ACTH, and IL-6 levels during melanoma growth <i>in vivo</i>.

<p>(A) GCR levels were measured by Western blot in control metastatic iB16 melanoma cells isolated from the liver and their equivalents stably expressing GCR-shRNA. Similar blots were run for B16-F10 and B16-F10-shGCR growing <i>in vitro</i>. Each lane in the blots corresponds to an individual representative animal in the indicated group. The relative density of each band was normalized against the internal standard (β-actin) on each blot (n = 4–5 in all cases) and expressed as relative changes in arbitrary densitometry units. Results obtained in cells transfected with lentiviral vector not harboring any gene (negative control) were not different from control values (not shown). *p<0.01 <i>versus</i> iB16 cells. <i>In vivo</i> experiments show data obtained after 7 days of inoculation. <i>In vitro</i> experiments show results obtained in cells cultured for 72h. (B–D)Blood was collected from the tail vein during a 24-h period starting 7 days after tumor inoculation, and peak plasma levels of corticosterone and ACTH (6 h and 12 h, circadian time, respectively) measured. Melanoma cells were isolated before GSH determination. Tumor volume and GSH levels were measured 8 days after inoculation. Data are mean values ± S.D. of 7–8 different animals. *p<0.05, **p<0.01 <i>versus</i> controls.</p

Effect of AS101 and anti-p53 antisense oligonucleotides on γ-GCS activity and expression and on GSH levels in metastatic melanoma cell subsets.

<p>Measurements and treatments were performed in isolated metastatic cells as indicated in the legend to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096466#pone-0096466-g005" target="_blank">Fig. 5</a>. Control experiments on p53 and Nrf2 levels were similar to those obtained in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096466#pone-0096466-g005" target="_blank">Fig. 5</a> A (not shown). Results obtained in iB16 cells transfected with p53 sense or scrambled oligonucleotides were not significantly different from those obtained in controls or cells incubated with AS101 alone (not shown). Data are mean values ± S.D. (n = 4–5 in all cases). *p<0.01 <i>versus</i> controls.</p

ROS, Nrf2 and GSH levels, and γ-GCS activity in iB16 and iB16-shGCR cells isolated from metastatic foci.

<p>Melanoma cells were isolated from the liver 7 days after inoculation, cultured, and transfected with anti-Nrf2-siRNA. H₂O₂ and O₂⁻generation, γ-GCS activity, and GSH levels were measured 48 h after seeding. Nrf2 levels (Western blotting) were measured 24 h after seeding. AU, arbitrary units. Data are mean values ± S.D. (n = 6–7 in all cases). *p<0.05,**p<0.01 <i>versus</i> iB16 controls. Results obtained in cells transfected with control Nrf2 sense or scrambled oligonucleotides were not significantly different from those obtained in cells cultured in the absence of anti-Nrf2-siRNA (not shown).</p