Cambios en la expresión de los genes involucrados en el ciclo celular y apoptosis durante el destete en la glándula mamaria de rata lactante. Papel del GSH.

RESUMEN En la glándula mamaria, al final de la lactancia, la acumulación de leche y el descenso de las hormonas lactogénicas inician una cascada de señalización que conduce a la muerte de las células secretoras por apoptosis y a la regresión de tejido mamario. Esta muerte celular se caracteriza por la liberación de citocromo c al citosol, donde activa las caspasas, responsables de los cambios observados en la apoptosis. Uno de ellos es la fragmentación del ADN internucleosomal, por activación de una endonucleasa. Además también existen cambios a nivel genético: el destete de 2 horas produce una inducción temprana de p53, mientras que otros genes como c-Jun o JNK no aumentan hasta 8 ó 24 horas de destete. La expresión y los niveles de proteína de p21cip1 y p27kip1, ambos inhibidores de las quinasas dependientes de ciclinas, que favorecen la detención del ciclo celular, también está significativamente aumentada tras el destete. Todos los cambios anteriores se reproducen en la glándula mamaria cuando disminuye la disponibilidad de GSH, bien con propargilglicina (PPG), un inhibidor de la transulfuración hepática con lo que disminuye la síntesis hepática de GSH y con ello su disponibilidad plasmática, o bien con butionina sulfoximina (BSO), inhibidor de la síntesis de GSH. Estos resultados sugieren que el flujo intertisular de GSH es un mecanismo muy importante para abastecer de L-cisteína a la glándula mamaria y resaltan la importancia de este proceso fisiológico en el mantenimiento de la lactancia. El óxido nítrico (NO) también está implicado en la apoptosis. A altas concentraciones forma derivados más reactivos que pueden inducir la apoptosis por: i) alteración mitocondrial; ii) activación de la vía de las MAP quinasas y iii) daño al ADN y acumulación de p53. En la glándula mamaria de rata lactante se expresan las tres isoformas de la enzima encargada de la síntesis de NO (NOS). No obstante, la expresión de la isoforma endotelial (eNOS) y de la inducible (iNOS) se modifican tras el destete o tras el tratamiento con BSO: los niveles de eNOS disminuyen tras un destete de 24 horas o tras el tratamiento con BSO, mientras que los de iNOS están aumentados con respecto al control y este incremento depende del tiempo de destete. Como consecuencia, la producción de NO en la glándula mamaria está aumentada durante la involución y este aumento induce la apoptosis y genera peroxinitrito, capaz de nitrar residuos de tirosina en las proteínas alterando su función. Podemos concluir que el NO juega un papel importante en la inducción de la apoptosis en la glándula mamaria. Por último, realizamos un estudio comparativo de los patrones de expresión génica en acini aislados del tejido mamario de ratas lactantes control, sometidas a un destete de 8 horas o tras el tratamiento con PPG mediante Microarray chip. De los, aproximadamente, 9000 genes comparados, sólo encontramos diferencias significativas en un 1-2% y establecimos dos patrones que se reproducían en ambas condiciones: el primero de ellos incluía genes implicados en la apoptosis celular: incrementaba la caspasa-6, la metaloproteinasa MMP-14, JunD o el antígeno CD14. El segundo patrón incluía enzimas implicados en el metabolismo glucídico y lipídico. Durante la lactancia la glándula mamaria es uno de los tejidos más activos en la captación de nutrientes; prácticamente el 50% de la glucosa captada será transformada en lípidos mediante la lipogénesis de novo. Sin embargo, en la involución se inhibe está síntesis de lípidos, por lo que los genes que codifican enzimas de esta vía disminuyen: piruvato deshidrogenasa, piruvato carboxilasa, acetil-CoA carboxilasa, ácido graso sintasa, etc. La disminución de GSH mimetiza los cambios encontrados durante el destete a nivel de la expresión génica. ____________________________________________________________________________________________________In the lactating mammary gland, weaning produces mitochondrial cytochrome c release and nuclear DNA fragmentation. This is followed by a significant decrease in milk production and the decline of lactation. Weaning for 2h produces an early induction of p53, whereas c-Jun and JNK are elevated after 24h of weaning. The expression of p21cip1 and p27kip1, both cyclin-dependent kinase inhibitors, is significantly higher in weaned rats when compared to control lactating rats. All the changes mentioned above also happen in the lactating mammary gland when PPG, an inhibitor of the liver trans-sulfuration pathway, is administered. This effect is partially reversed by N-acetylcysteine (NAC) administration. The administration of buthionine sulfoximine (BSO), an irreversible inhibitor of -glutamyl cysteine synthetase, to lactating rats produces a decrease in GSH levels and changes in protein levels and gene transcripts similar to the rats with impaired trans-sulfuration pathway. These data suggest that the intertissue flux of glutathione is an important mechanism of L-cysteine delivery to the lactating mammary gland and emphasizes the importance of this physiological event to maintain gene expression required to keep lactation. Once characterized the apoptosis in the mammary gland we used this model to study in vivo the role of nitric oxide during cell death. The three isoforms of the nitric oxide synthase (NOS) are constitutively present in the acini of the lactating mammary gland. The amount of iNOS and NO production are increased during weaning when compared to control lactating glands, but the protein levels of the eNOS isoform are diminished. Western blot analysis demonstrated that protein nitration was increased in the weaned mammary gland, althought limited to a few specific tyrosine-nitrated proteins. A decrease of GSH in the mammary tissue at the peak of lactation partially mimics these findings and emphasizes the role of NO as a potential signal that triggers involution

Improving childhood obesity treatment using new technologies: the ETIOBE System

Childhood obesity is an increasing public health problem in western culture. Sedentary lifestyles and an “obesogenic environment” are the main influences on children leading to an increase in obesity. The objective of this paper is to describe an e-health platform for the treatment and prevention of childhood obesity called ETIOBE. This e-health platform is an e-therapy system for the treatment of obesity, aimed at improving treatment adherence and promoting the mechanisms of self-control in patients, to obtain weight loss maintenance and to prevent relapse by establishing healthy lifestyle habits. ETIOBE is composed of three different applications, the Clinician Support System (CSS), the Home Support System (HSS) and the Mobile Support System (MSS). The use of new Information and Communication (ICT) technologies can help clinicians to improve the effectiveness of weight loss treatments, especially in the case of children, and to achieve designated treatment goal

Calpains, the proteases of two faces controlling the epithelial homeostasis in mammary gland

Calpain-1 and calpain-2 are calcium-dependent Cys-proteases ubiquitously expressed in mammalian tissues with a processive, rather than degradative activity. They are crucial for physiological mammary gland homeostasis as well as for breast cancer progression. A growing number of evidences indicate that their pleiotropic functions depend on the cell type, tissue and biological context where they are expressed or dysregulated. This review considers these standpoints to cover the paradoxical role of calpain-1 and -2 in the mammary tissue either, under the physiological conditions of the postlactational mammary gland regression or the pathological context of breast cancer. The role of both calpains will be examined and discussed in both conditions, followed by a brief snapshot on the present and future challenges for calpains, the two-gateway proteases towards tissue homeostasis or tumor development

Vitamin A Status Modulates Epithelial Mesenchymal Transition in the Lung: The Role of Furin

Vitamin A deficiency (VAD) induced TGF-β hyperactivation and reduced expression of cell adhesion proteins in the lung, suggesting that the disruption of retinoic acid (RA) signaling leads to epithelial-mesenchymal transition (EMT). To elucidate the role of lung vitamin A status in EMT, several EMT markers and the expression of the proprotein convertase furin, which activates TGF-β, were analyzed in two experimental models. Our in vivo model included control rats, VAD rats, and both control rats and VAD rats, treated with RA. For the in vitro studies, human bronchoalveolar epithelial cells treated with RA were used. Our data show that EMT and furin are induced in VAD rats. Furthermore, furin expression continues to increase much more markedly after treatment of VAD rats with RA. In control rats and cell lines, an acute RA treatment induced a significant increase in furin expression, concomitant with changes in EMT markers. A ChIP assay demonstrated that RA directly regulates furin transcription. These results emphasize the importance of maintaining vitamin A levels within the physiological range since both levels below and above this range can cause adverse effects that, paradoxically, could be similar. The role of furin in EMT is discussed

PDA self-register system for eating disorders: a study on acceptability and satisfaction

Self-monitoring techniques, such as the use of dietary registers, are considered to be central to cognitive-behavioral treatment of Eating Disorders (ED). This information allows the clinician to identify the triggers of the behaviors associated to ED as purges and/or binges, and the associated thoughts and emotions, helping to carry out a more accurate assessment. Traditionally these registers are made with paper and pencil mode, where the patient has to register every eating and the emotions/thoughts associated; but this system has some problems, as low portability, low adherence or methodological difficulties. The use of PDA for self-registers can help to solve these problems. The aim of this study is to study the levels of acceptability and satisfaction with PDAs self-register system specifically designed for assessment and treatment of ED. Samples of 30 subjects diagnosed with ED are receiving a PDA with software specifically designed for recording type and amount of food, emotions before and after eating and other behaviors. The participants are completing self-register daily during a week, and afterwards answer an acceptance and satisfaction questionnaire. This work is in progress at the moment. It is expected that the PDA system will show high levels of acceptance and satisfaction

Cationic amino acid transport across the blood-brain barrier is mediated exclusively by system y+

Cationic amino acid (CAA) transport is brought about by two families of proteins that are found in various tissues: Cat (CAA transporter), referred to as system y+, and Bat [broad-scope amino acid (AA) transporter], which comprises systems b0,+, B0,+, and y+L. CAA traverse the blood-brain barrier (BBB), but experiments done in vivo have only been able to examine the BBB from the luminal (blood-facing) side. In the present study, plasma membranes isolated from bovine brain microvessels were used to identify and characterize the CAA transporter(s) on both sides of the BBB. From these studies, it was concluded that system y+ was the only transporter present, with a prevalence of activity on the abluminal membrane. System y+ was voltage dependent and had a Km of 470 ± 106 μM (SE) for lysine, a Ki of 34 μM for arginine, and a Ki of 290 μM for ornithine. In the presence of Na+, system y+ was inhibited by several essential neutral AAs. The Ki values were 3–10 times the plasma concentrations, suggesting that system y+ was not as important a point of access for these AAs as system L1. Several small nonessential AAs (serine, glutamine, alanine,and glycine) inhibited system y+ with Ki values similar to their plasma concentrations, suggesting that system y+ may account for the permeability of the BBB to these AAs. System y+ may be important in the provision of arginine for NO synthesis. Real-time PCR and Western blotting techniques established the presence of the three known nitric oxide synthases in cerebral endothelial cells: NOS-1 (neuronal), NOS-2 (inducible), and NOS-3 (endothelial). These results confirm that system y+ is the only CAA transporter in the BBB and suggest that NO can be produced in brain endothelial cells

Allocated zones for aquaculture : a guide for the establishment of coastal zones dedicated to aquaculture in the Mediterranean and the Black Sea

This publication is based on the findings of the project Developing site selection and carrying capacity guidelines for Mediterranean aquaculture within aquaculture appropriate areas (SHoCMed), implemented by the Scientific Advisory Committee on Aquaculture (CAQ) of the General Fisheries Commission for the Mediterranean (GFCM) of the Food and Agriculture Organization of the United Nations (FAO) with the support of the European Union (Directorate-General for Maritime Affairs and Fisheries of the European Commission). Concluded in 2016, SHoCMed aimed at developing criteria, standards and guidelines for GFCM contracting parties and cooperating non-contracting parties to improve site selection criteria and set capacity benchmarks and references for aquaculture activities in the Mediterranean.peer-reviewe

Electronic PDA dietary and physical activity registers in a weight loss treatment program for children: A description of the ETIOBE Personal Digital Assistant System

[EN] Childhood obesity is a significant health problem in western societies. Self-monitoring techniques, such as the use of dietary and physical activity registers, are considered to be central to cognitive-behavioral weight control programs. Traditionally, these conventional diaries have been created using pen and paper, however, this technique has several limitations. The objective of this paper is to describe an electronic Personal Digital Assistant (PDA) system for recording food and physical activity for the treatment of childhood obesity. The authors review the benefits and limitations of such electronic diaries.This study was funded in part by Ministerio de Educación y Ciencia Spain, Projects Consolider-C (SEJ2006-14301/PSIC), CIBER of Physiopathology of Obesity and Nutrition, an initiative of ISCIII and Excellence Research Program PROMETEO (Generalitat Valenciana. Conselleria de Educación, 2008-157).Baños, RM.; Cebolla, A.; Zaragozá, I.; Botella, C.; Alcañiz Raya, ML. (2009). Electronic PDA dietary and physical activity registers in a weight loss treatment program for children: A description of the ETIOBE Personal Digital Assistant System. JOURNAL OF CYBERTHERAPY & REHABILITATION. 2(3):235-241. http://hdl.handle.net/10251/1504482352412