Purinergic P2Y2 and P2X4 Receptors Are Involved in the Epithelial-Mesenchymal Transition and Metastatic Potential of Gastric Cancer Derived Cell Lines.

Gastric cancer (GC) is a major health concern worldwide, presenting a complex pathophysiology that has hindered many therapeutic efforts so far. In this context, purinergic signaling emerges as a promising pathway for intervention due to its known role in cancer cell proliferation and migration. In this work, we explored in more detail the role of purinergic signaling in GC with several experimental approaches. First, we measured extracellular ATP concentrations on GC-derived cell lines (AGS, MKN-45, and MKN-74), finding higher levels of extracellular ATP than those obtained for the non-tumoral gastric cell line GES-1. Next, we established the P2Y2 and P2X4 receptors (P2Y2R and P2X4R) expression profile on these cells and evaluated their role on cell proliferation and migration after applying overexpression and knockdown strategies. In general, a P2Y2R overexpression and P2X4R downregulation pattern were observed on GC cell lines, and when these patterns were modified, concomitant changes in cell viability were observed. These modifications on gene expression also modified transepithelial electrical resistance (TEER), showing that higher P2Y2R levels decreased TEER, and high P2X4R expression had the opposite effect, suggesting that P2Y2R and P2X4R activation could promote and suppress epithelial-mesenchymal transition (EMT), respectively. These effects were confirmed after treating AGS cells with UTP, a P2Y2R-agonist that modified the expression patterns towards mesenchymal markers. To further characterize the effects of P2Y2R activation on EMT, we used cDNA microarrays and observed that UTP induced important transcriptional changes on several cell processes like cell proliferation induction, apoptosis inhibition, cell differentiation induction, and cell adhesion reduction. These results suggest that purinergic signaling plays a complex role in GC pathophysiology, and changes in purinergic balance can trigger tumorigenesis in non-tumoral gastric cells.

Differential Effects of Purinergic Signaling in Gastric Cancer-Derived Cells Through P2Y and P2X Receptors.

Gastric cancer (GC) is the one of the most prevalent cancers and one of the leading causes of cancer-induced deaths. Previously, we found that the expression of purinergic P2Y2 receptor (P2Y2R) is increased in GC samples as compared to adjacent healthy mucosa taken from GC-diagnosed patients. In this work, we studied in detail purinergic signaling in the gastric adenocarcinoma-derived cell lines: AGS, MKN-45, and MKN-74, and compared them to a nontumoral epithelial cell line: GES-1. In GC-derived cells, we detected the expression of several purinergic receptors, and found important differences as compared to GES-1 cells. Functional studies revealed a strong contribution of P2Y2Rs in intracellular calcium increases, elicited by adenosine-triphosphate (ATP), uridine-triphosphate (UTP), and the P2Y2R agonist MRS2768. Responses were preserved in the absence of extracellular calcium and inhibited by P2Y2R antagonists. In GES-1 cells, ATP and UTP induced similar responses and the combination of P2X and P2Y receptor antagonists was able to block them. Proliferation studies showed that ATP regulates AGS and MKN-74 cells in a biphasic manner, increasing cell proliferation at 10-100 µM, but inhibiting at 300 µM ATP. On the other hand, 1-300 µM UTP, a P2Y2R agonist, increased concentration-dependent cell proliferation. The effects of UTP and ATP were prevented by both wide-range and specific purinergic antagonists. In contrast, in GES-1 cells ATP only decreased cell proliferation in a concentration-dependent manner, and UTP had no effect. Notably, the isolated application of purinergic antagonists was sufficient to change the basal proliferation of AGS cells, indicating that nucleotides released by the cells can act as paracrine/autocrine signals. Finally, in tumor-derived biopsies, we found an increase of P2Y2R and a decrease in P2X4R expression; however, we found high variability between seven different biopsies and their respective adjacent healthy gastric mucosa. Even so, we found a correlation between the expression levels of P2Y2R and P2X4R and survival rates of GC patients. Taken together, these results demonstrate the involvement of different purinergic receptors and signaling in GC, and the pattern of expression changes in tumoral cells, and this change likely directs ATP and nucleotide signaling from antiproliferative effects in healthy tissues to proliferative effects in cancer.

Nitrogen physiology of contrasting genotypes of Chenopodium quinoa Willd. (Amaranthaceae).

Quinoa has been highlighted as a promising crop to sustain food security. The selection of physiological traits that allow identification genotypes with high Nitrogen use efficiency (NUE) is a key factor to increase Quinoa cultivation. In order to unveil the underpinning mechanisms for N-stress tolerance in Quinoa, three genotypes with similar phenology, but different NUE were developed under high (HN) or low (LN) nitrogen conditions. N metabolism processes and photosynthetic performance were studied after anthesis and in correlation with productivity to identify principal traits related to NUE. We found that protein content, net photosynthesis and leaf dry-mass were determinant attributes for yield at both HN and LN conditions. Contrastingly, the enhancement of N related metabolites ([Formula: see text], proline, betacyanins) and processes related with re-assimilation of [Formula: see text], including an increment of glutamine synthetase activity and up-regulation of CqAMT1,1 transporter expression in leaves, were negatively correlated with grain yield at both N conditions. Biochemical aspects of photosynthesis and root biomass were traits exclusively associated with grain yield at LN. The impact of N supply on seed quality is discussed. These results provide new insights towards the understanding the N metabolism of Quinoa.

Nitrogen Supply Affects Photosynthesis and Photoprotective Attributes During Drought-Induced Senescence in Quinoa.

Drought during senescence has become more common in Mediterranean climates in recent years. Chenopodium quinoa Willd has been identified as tolerant to poor soil conditions and drought. Previous observations have found that sufficient nitrogen (N) supply mitigates yield losses under terminal drought conditions. However, there is no understanding of the mechanisms behind this effect. We hypothesized that N up-regulates both photosynthetic and photoprotective elements during drought-induced senescence, alleviating the negative impact of drought on yield. The role of N supply and terminal drought on photoprotection was tested using three Chilean quinoa genotypes from different climatic zones: Faro, UdeC9, and BO78. Plants were grown under high nitrogen (HN) or low nitrogen (LN) conditions and subjected to terminal drought at the onset of senescence. Photosynthetic and photochemical and non-photochemical processes were evaluated at both the onset of drought and after 15 days of drought conditions. N supplementation modified most of the physiological parameters related to photochemical dissipation of energy, photosynthesis, and yield in quinoa. In contrast, water restriction did not affect photosynthesis in quinoa, and its effect on yield was dependent on the genotype. A significant interaction N × G was observed in photosynthesis, relative water content, protein content, Fv/Fm, and chlorophylls. In general, Faro was able to maintain higher levels of these attributes under LN conditions than UdeC9 and BO78. In addition, the interacting effects of N × W regulated the level of most pigments in quinoa as well as the photoprotective induction of non-photochemical quenching (NPQ) during senescence. During terminal drought at LN conditions, Faro presented a larger NPQ induction under drought conditions than UdeC9 and BO78, which was supported by a larger zeaxanthin content and de-epoxidation state of the xanthophyll pool. Interestingly, BO78 did not induce NPQ in response to drought-induced senescence but instead enhanced the content of betacyanins. This response needs to be researched in future works. Finally, we observed that LN supply reduced the correlationship between the de-epoxidation state of the xanthophyll cycle and NPQ. This could be an indication that N supply not only compromised the capacity for photosynthetic performance in quinoa plants, but also affected the plasticity of thermal dissipation, restricting further changes during drought-induced senescence.

The impact of different agroecological conditions on the nutritional composition of quinoa seeds.

Quinoa cultivation has been expanded around the world in the last decade and is considered an exceptional crop with the potential of contributing to food security worldwide. The exceptional nutritional value of quinoa seeds relies on their high protein content, their amino acid profile that includes a good balance of essential amino acids, the mineral composition and the presence of antioxidants and other important nutrients such as fiber or vitamins. Although several studies have pointed to the influence of different environmental stresses in certain nutritional components little attention has been paid to the effect of the agroecological context on the nutritional properties of the seeds what may strongly impact on the consumer food's quality. Thus, aiming to evaluate the effect of the agroecological conditions on the nutritional profile of quinoa seeds we analyzed three quinoa cultivars (Salcedo-INIA, Titicaca and Regalona) at different locations (Spain, Peru and Chile). The results revealed that several nutritional parameters such as the amino acid profile, the protein content, the mineral composition and the phytate amount in the seeds depend on the location and cultivar while other parameters such as saponin or fiber were more stable across locations. Our results support the notion that nutritional characteristics of seeds may be determined by seed's origin and further analysis are needed to define the exact mechanisms that control the changes in the seeds nutritional properties.

Factores de riesgo en los trastornos de conducta alimentaria en alumnos de las Facultades de Medicina Humana y Obstetricia y Puericultura de la Universidad Católica de Santa María - Arequipa 2008 / Risk factors in eating disorders among students of the Faculty of Human Medicine and Midwifery at the Catholic University of Santa Maria - Arequipa 2008

Introducción: El presente estudio pretende determinar los trastornos de conducta alimentaria en los alumnos de las facultades mencionadas y establecer la relación entre los factores de riesgo y estas patologías. Método: Se aplicó 164 cuestionarios en el que se recolectan las características generales de la población y el Eating Disorders Inventory (EDI), de los cuales 4 corresponden a la facultad de Obstetricia y Puericultura y 100 a la facultad de de Medicina Humana. Resultados: Se encontró que la mayor parte de los participantes en el estudio fueron de sexo femenino con una edad promedio de 18 años de edad, mientras que para el sexo masculino fue de 19 años. El 35 por ciento de estudiantes a los que se les aplicó el cuestionario se encuentran en riesgo de padecer trastornos de conducta alimentaria, mientras que un 65 por ciento de ellos no pueden ser catalogados como sujetos de riesgo. Conclusiones: Dentro de los factores de riesgo, la obsesión por la delgadez es el mayor con un 25.86 por ciento, cabe resaltar que este es el síntoma principal para diagnosticar los casos de anorexia y bulímia.