Assessment of different pre-treatment methods for the removal of limonene in citrus waste and their effect on methane potential and methane production rate

The objective of this study was to assess the limonene removal efficiency of three pre-treatment methods when applied to citrus waste and to evaluate their effects on the biochemical methane potential (BMP) and the methane production rate (MPR) using batch anaerobic testsPostprint (published version

Preclinical efficacy of [V 4 Q 5 ]dDAVP, a second generation vasopressin analog, on metastatic spread and tumor-associated angiogenesis in colorectal cancer

Purpose Control of metastatic spread of colorectal cancer (CRC) remains as a major therapeutic challenge. [V 4 Q 5 ]dDAVP is a vasopressin peptide analog with previously reported anticancer activity against carcinoma tumors. By acting as a selective agonist of arginine vasopressin type 2 membrane receptor (AVPR2) present in endothelial and tumor cells, [V 4 Q 5 ]dDAVP is able to impair tumor aggressiveness and distant spread. Our aim was to evaluate the potential therapeutic benefits of [V 4 Q 5 ]dDAVP on highly aggressive CRC disease using experimental models with translational relevance. Materials and Methods Murine CT-26 and human Colo-205 AVPR2-expressing CRC cell lines were used to test the preclinical efficacy of [V 4 Q 5 ]dDAVP, both in vitro and in vivo. Results In syngeneic mice surgically implanted with CT-26 cells in the spleen, sustained intravenous treatment with [V 4 Q 5 ]dDAVP (0.3 jg/kg) dramatically impaired metastatic progression to liver without overt signs of toxicity, and also reduced experimental lung colonization. The compound inhibited in vivo angiogenesis driven by Colo-205 cells in athymic mice, as well as in vitro endothelial cell migration and capillary tube formation. [V 4 Q 5 ]dDAVP exerted AVPR2-dependent cytostatic activity in vitro (IC50 1.08 |jM) and addition to 5-fluorouracil resulted in synergistic antiproliferative effects both in CT-26 and Colo-205 cells. Conclusion The present preclinical study establishes for the first time the efficacy of [V 4 Q 5 ]dDAVP on CRC. These encouraging results suggest that the novel second generation vasopressin analog could be used for the management of aggressive CRC as an adjuvant agent during surgery or to complement standard chemotherapy, limiting tumor angiogenesis and metastasis and thus protecting the patient from CRC recurrence.Fil: Garona, Juan. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sobol, Natasha Tatiana. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pifano, Marina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Segatori, Valeria Inés. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gomez, Daniel Eduardo. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ripoll, Giselle Vanina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Alonso, Daniel Fernando. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

La medición del tiempo de trabajo reproductivo a través de la Encuesta de Empleo del Tiempo y la Encuesta Nacional de Salud: un análisis comparativo

Comunicación presentada en el XI Congreso Español de Sociología, Madrid, 10-12 julio 2013.La medición de la dedicación al trabajo no remunerado permite visibilizar la relevancia de este tipo de actividades y su aportación al bienestar económico y social. Con la finalidad de contribuir al conocimiento sobre la medición y cuantificación de las actividades no remuneradas, la presente propuesta consiste en el análisis comparativo de las formas de medición del tiempo de trabajo no remunerado entre la Encuesta de Empleo del Tiempo 2002-2003 del INE y la Encuesta Nacional de Salud 2006 del INE-Ministerio de Sanidad y Consumo, concretamente, del tiempo dedicado a las tareas domésticas, al cuidado de personas adultas miembros del hogar y al cuidado de menores, atendiendo a las diferencias de género. Se trata de dos encuestas de representatividad nacional. La EET 2002-2003 incluye a una muestra de 46.774 individuos de 10 y más años y la ENS2006 a 29.478 personas de 16 y más años. Para nuestro análisis se utilizó la población de 18 y más años. El cuanto al análisis estadístico empleado, se han calculado las medias del tiempo diario dedicado a cada tipo de trabajo en ambas encuestas y, para la comparación de las mediciones, se han realizado contrastes estadísticos de la diferencia de medias para muestras independientes, con un intervalo de confianza al 95%. Los resultados del análisis muestra las divergencias en la estimación del tiempo diario dedicado al trabajo reproductivo entre las dos fuentes estadísticas utilizadas, sobre todo en el cuidado informal. El origen de estas diferencias se halla tanto en las distintas definiciones de las personas cuidadoras y de las personas que precisan de cuidados como, principalmente, en los diferentes instrumentos utilizados para la recogida de la información

The impacts of providing family care on caregivers' daily life

El objetivo del artículo es analizar la distribución del tiempo de las personas cuidadoras en las distintas esferas de la vida (la esfera laboral, la privada o personal y la doméstica). El estudio se basa en el análisis de los datos de la Encuesta de Empleo del Tiempo 2002-2003 del Instituto Nacional de Estadística (INE). Los resultados de la investigación muestran los impactos negativos del cuidado informal sobre los distintos ámbitos de la vida en términos de reducción de la vida privada o personal, de la participación en el ámbito laboral y de mayor carga del trabajo total. Asimismo, también ponen de manifiesto la heterogeneidad de los efectos del cuidado informal en la vida cotidiana de las personas cuidadoras según el sexo, la edad y el nivel de ingresos.The aim of this paper is to analyse the use of time of caregivers in their life spheres (the labour sphere, personal life and housework). The study is based on the analysis of data from the Spanish Time Budget Survey 2002-2003 carried out by the National Statistics Institute. The results show informal care negative impacts in terms of personal time reduction, decrease participation in the labour market and increased overall time devoted to unpaid and paid work. Also, the results show the different impacts of informal care on caregivers’ daily life according to gender, age and household income level.El presente trabajo forma parte del proyecto de investigación «Diversidad de género, clase social y etnia en la atención a la dependencia. Las personas cuidadoras y dependientes» (Expediente: MD07/00109), financiado por el Instituto de Salud Carlos III, Ministerio de Sanidad y Consumo, y dirigido por Daniel La Parra Casado

Use of 13Ca chemical-shifts in protein structure determination

A physics-based method aimed at determining protein structures by using NOE-derived distances together with observed and computed 13C chemical shifts is proposed. The approach makes use of 13Cα chemical shifts, computed at the density functional level of theory, to obtain torsional constraints for all backbone and side-chain torsional angles without making a priori use of the occupancy of any region of the Ramachandran map by the amino acid residues. The torsional constraints are not fixed but are changed dynamically in each step of the procedure, following an iterative self-consistent approach intended to identify a set of conformations for which the computed 13Cα chemical shifts match the experimental ones. A test is carried out on a 76-amino acid, all-α-helical protein; namely, the Bacillus subtilis acyl carrier protein. It is shown that, starting from randomly generated conformations, the final protein models are more accurate than an existing NMR-derived structure model of this protein, in terms of both the agreement between predicted and observed 13Cα chemical shifts and some stereochemical quality indicators, and of similar accuracy as one of the protein models solved at a high level of resolution. The results provide evidence that this methodology can be used not only for structure determination but also for additional protein structure refinement of NMR-derived models deposited in the Protein Data Bank.Fil: Vila, Jorge Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Matemática Aplicada de San Luis "Prof. Ezio Marchi". Universidad Nacional de San Luis. Facultad de Ciencias Físico, Matemáticas y Naturales. Instituto de Matemática Aplicada de San Luis "Prof. Ezio Marchi"; Argentina. Cornell University; Estados UnidosFil: Ripoll, Daniel R.. Cornell Theory Center; Estados UnidosFil: Scheraga, Harold A.. Cornell University; Estados Unido

Biomedical Applications of Tissue Clearing and Three-Dimensional Imaging in Health and Disease.

Three-dimensional (3D) optical imaging techniques can expand our knowledge about physiological and pathological processes that cannot be fully understood with 2D approaches. Standard diagnostic tests frequently are not sufficient to unequivocally determine the presence of a pathological condition. Whole-organ optical imaging requires tissue transparency, which can be achieved by using tissue clearing procedures enabling deeper image acquisition and therefore making possible the analysis of large-scale biological tissue samples. Here, we review currently available clearing agents, methods, and their application in imaging of physiological or pathological conditions in different animal and human organs. We also compare different optical tissue clearing methods discussing their advantages and disadvantages and review the use of different 3D imaging techniques for the visualization and image acquisition of cleared tissues. The use of optical tissue clearing resources for large-scale biological tissues 3D imaging paves the way for future applications in translational and clinical research.This work was supported by Ministerio de Ciencia, Innovacion y Universidades, ISCIII-FIS grants PI18/00462 co-financed by ERDF, European Union (FEDER) Funds from the European Commission, European Union, ‘‘A way of making Europe’’; the CNIC is supported by theMinisterio de Ciencia, Innovacion y Universidades y the Pro CNIC Foundation, Severo Ochoa Center of Excellence (SEV-2015-0505), CIBER de Salud Mental (CIBERSAM), and COST-action CA16124. J.R. acknowledges funding from EU H2020 FET Open project SENSITIVE, ID 801347, and Ministerio de Ciencia, Innovacion y Universidades Grant FIS2016-77892-RS

Commentary: Arginine vasopressin receptor 1a is a therapeutic target for castration-resistant prostate cancer

Prostate cancer patients managed with androgen-deprivation therapy usually recur after a few years and the disease gradually becomes castration-resistant prostate cancer (CRPC). The role of tumor cell plasticity, including processes such as transdifferentiation and epithelial-mesenchymal transition, is pivotal in the development of androgen receptor (AR)-indifferent tumor variants (1). Cell plasticity may allow CRPC progression and metastasis by favoring reactivation of AR signaling as a result of different mechanisms of transcriptome reprogramming. Interestingly, dissection of such mechanisms can lead to the identification of novel vulnerabilities of aggressive tumor cells that can be targeted therapeutically. The recent work by Zhao et al. (2) identified the vasopressin receptor 1a (AVPR1a) as a critical effector in CRPC expressing the AR coactivator VAV3 and the constitutively active AR variant AR-V7. They demonstrated that ectopic expression of AVPR1a is capable of conferring castration resistance and agonist treatment with the receptor ligand, the natural hormone arginine vasopressin, activates ERK and CREB, signaling molecules known to promote prostate cancer progression. Interestingly, depletion of AVPR1a or inhibition by the selective AVPR1a antagonist relcovaptan resulted in decreased CRPC cell proliferation and reduced bone metastatic growth in vivo. We completely agree with the authors in the sense that AVPR1a can be a potential target for CRPC therapy. We believe that clinical trials with relcovaptan are warranted, particularly in patients with bone-metastatic disease for which therapeutic options are limited. However, we want to point out that these results could be revealing other untapped antitumor properties of the vasopressin system-related drugs against prostate cancer cells. Our team has reported that the vasopressin analog desmopressin, a selective agonist for the vasopressin receptor 2 (AVPR2), significantly reduced tumor cell growth and migration in AR-negative CRPC (3). In vitro exposure to desmopressin also induced a dramatic decrease of the neuroendocrine markers chromogranin and neuron-specific enolase in aggressive CRPC cells (3). In prostate cancer, neuroendocrine transdifferentiation is known to be related with transition toward AR-indifference and metastatic phenotype. Besides, recent studies in orthotopic and heterotopic models of CRPC in athymic nude mice demonstrated an enhanced efficacy of docetaxel in combination with desmopressin (4, 5). Agonist activation of AVPR2 present in various human cancer cell lines has been associated with triggering of antiproliferative signaling pathways involving canonical adenylate cyclase/cAMP/PKA axis activation. cAMP blocks the proliferation of many cell types, both normal and transformed, through multiple downstream effectors. It is known that increased cAMP levels inhibit the Raf/MAPK/ERK signaling pathway in a PKA-dependent manner, but this is not the only mitogenic pathway impaired by cAMP (6). Although cytostatic effect after AVPR2 stimulation is robust, the underlying mechanism is intriguing since AVPR2-mediated cell signaling may eventually lead to phosphorylation of hundreds of PKA substrates granting a complex and seemingly contradictory framework of signaling pathways as evidenced in immortalized epithelial cells after stimulation with desmopressin (7). In this sense, high cAMP levels and PKA activation may eventually lead to phosphorylation and activation of CREB. However, many factors contribute to final cell response. It is known that cAMP can either stimulate or inhibit tumor cell apoptosis, and PKA is able to mediate cAMP-promoted proapoptotic responses depending on the cell type and the cell context (8). In addition, it has been reported a crosstalk of cAMP/PKA that can inhibit RhoA-mediated signaling, thus affecting the aggressive behavior of CRPC cells (9). Taken together, it seems that CREB activation is not prominent after AVPR2 selective agonist action on cancer cells, being favored certain antiproliferative signals. In breast cancer cells expressing AVPR1a and AVPR2, natural vasopressin can activate both receptors but its affinity is higher for AVPR1a and the number of functional AVPR2 tends to be relatively low. Thus, vasopressin elicits AVPR1a-dependent proliferative signals mediated by ERK activation that clearly predominate over AVPR2-dependent antiproliferative signals (10). On the contrary, when AVPR1a is blocked by selective antagonists such as relcovaptan or tumor cells are exposed to specific AVPR2 agonists such as desmopressin, significant antiproliferative effects are achievable in hormone-resistant breast cancer cell lines (10, 11). Desmopressin also contributed to reduce aggressiveness of mammary tumors during chemotherapy in an immunocompetent mouse model (12). We have conducted a Phase 2 dose-escalation clinical trial of desmopressin as a perioperative adjuvant in patients with breast cancer (NCT01606072). Desmopressin appeared safe when administered in two slow infusions before and after surgery, and a rapid postoperative drop in circulating tumor cells was detected after treatment (13). In addition, we documented a reduced intraoperative bleeding associated to the well-known hemostatic effects of the compound (13). From a wider perspective, the stimulating article by Zhao et al. (2) ratifies the relevance of the vasopressin system for searching novel therapeutic targets in hormone-resistant cancer and particularly in CRPC (Figure 1; see also Supplementary Table 1 for preclinical data summary). In this context, repurposing of already-used drugs with a non-oncology primary purpose stands as an interesting strategy to offer effective therapeutic options to cancer patients, allowing faster development and reducing safety concerns (14). The selective AVPR1a antagonist relcovaptan is a small-molecule inhibitor that has been safely and effectively used in clinical trials for Raynaud syndrome, dysmenorrhea and preterm labor. The specific AVPR2 agonist desmopressin is a synthetic peptide compound that has been employed for decades as an antidiuretic in the treatment of diabetic insipidus and enuresis, and as a hemostatic agent for the management of bleeding disorders, with a history of good tolerability and clinical effectiveness. Both compounds constitute promising therapeutic approaches for CRPC that deserve clinical testing either alone or in combination, as well as concurrently with standard chemotherapy regimens.Fil: Ripoll, Giselle Vanina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pifano, Marina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Garona, Juan. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Alonso, Daniel Fernando. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Effects of the synthetic vasopressin analog desmopressin in a mouse model of colon cancer

Experimental and clinical data indicated that perioperative administration of the hemostatic peptide desmopressin (DDAVP) can inhibit progression of residual metastatic cells. The compound seems to act by inducing an agonist effect on specific V2 vasopressin membrane receptors present in both tumor cells and endothelial cells. Here we explored the antitumor effects of DDAVP in cultured colon carcinoma cells and in a syngeneic Balb/c mouse model. Both human Colo-205 and mouse CT-26 colon carcinoma cell lines expressed the V2 receptor, as revealed by immunofluorescence. DDAVP (at doses ranging from 100 ng/ml to 1 μg/ml) exerted a modest but significant antiproliferative effect on cultured CT26 and Colo-205 cells. In vivo, DDAVP (2 intravenous doses of 2 μg/kg) reduced accumulation of ascites and formation of intestinal tumor nodules in mice intraperitoneally inoculated with CT-26 cells. Perioperative administration of DDAVP significantly inhibited tumor progression in animals surgically implanted in the spleen with CT-26 cells, and caused some reduction in liver metastasis. Although DDAVP and 5- fluorouracil demonstrated additive cytostatic effects in vitro, no antitumor effects were observed in this study in mice receiving a single cycle of chemotherapy (25 mg/kg) in combination with the peptide. Our data suggest that DDAVP may be potentially used to minimize spread or survival of residual malignant cells during surgical procedures for colon and other gastrointestinal tumors.Fil: Ripoll, Giselle Vanina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Garona, Juan. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Hermo, Guillermo Adrian. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gomez, Daniel Eduardo. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Alonso, Daniel Fernando. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin