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Background: The ''predictD algorithm'' provides an estimate of the level and profile of risk of the onset of major depression in primary care attendees. This gives us the opportunity to develop interventions to prevent depression in a personalized way. We aim to evaluate the effectiveness, cost-effectiveness and cost-utility of a new intervention, personalized and implemented by family physicians (FPs), to prevent the onset of episodes of major depression.Methods/Design: This is a multicenter randomized controlled trial (RCT), with cluster assignment by health center and two parallel arms. Two interventions will be applied by FPs, usual care versus the new intervention predictD-CCRT. The latter has four components: a training workshop for FPs; communicating the level and profile of risk of depression; building up a tailored bio-psycho-family-social intervention by FPs to prevent depression; offering a booklet to prevent depression; and activating and empowering patients. We will recruit a systematic random sample of 3286 non-depressed adult patients (1643 in each trial arm), nested in 140 FPs and 70 health centers from 7 Spanish cities. All patients will be evaluated at baseline, 6, 12 and 18 months. The level and profile of risk of depression will be communicated to patients by the FPs in the intervention practices at baseline, 6 and 12 months. Our primary outcome will be the cumulative incidence of major depression (measured by CIDI each 6 months) over 18 months of follow-up. Secondary outcomes will be health-related quality of life (SF-12 and EuroQol), and measurements of cost-effectiveness and cost-utility. The inferences will be made at patient level. We shall undertake an intention-to-treat effectiveness analysis and will handle missing data using multiple imputations. We will perform multi-level logistic regressions and will adjust for the probability of the onset of major depression at 12 months measured at baseline as well as for unbalanced variables if appropriate. The economic evaluation will be approached from two perspectives, societal and health system.Discussion: To our knowledge, this will be the first RCT of universal primary prevention for depression in adults and the first to test a personalized intervention implemented by FPs. We discuss possible biases as well as other limitations.Trial registration: ClinicalTrials.gov identifier: NCT01151982.

The lectin Reg3b provides crucial protection to various tissues against inflammation, a potential risk factor for pancreatic ductal adenocarcinoma. Reg3b is also overexpressed in serum and pancreatic juice from patients with this cancer, but its function in this context remains to be elucidated. In this study, we investigated the role of Reg3b in tumor development in an orthotopic mouse model of pancreatic cancer. Reg3b deletion in mice drastically impaired pancreatic tumor growth, correlating with decreased angiogenesis and increased apoptosis of tumor cells. Moreover, Reg3b deficiency resulted in an alteration of the tumoral immune microenvironment, reflected by a decrease in the M2/M1 ratio of tumor-associated macrophages and an upregulation of CD3 cell infiltration. Addition of Reg3b to prestimulated RAW 264.7 or primary macrophages enhanced M2 polarization through the activation of STAT3 signaling pathway. Conditioned media from Reg3b-M2-polarized primary macrophages inhibited apoptosis and prolonged the viability of Panc02 tumor cells. Our studies reveal a novel role for Reg3b as a tumor promoter in pancreatic adenocarcinoma through the regulation of tumor stroma. Thus, inhibition of this protein may be a useful strategy in treatment of pancreatic cancer. © 2013 American Association for Cancer Research., This work was supported by FIS grants from Instituto de Salud Carlos III (PI081608 to E. Folch-Puy, PI1200519 to J. Rosello-Catafau, and PI1002888 to M. Gironella; cofunded by FEDER-European Union). CIBEREHD is funded by the Instituto de Salud Carlos III.

Recently, constitutional MLH1 epimutations have been identified in a subset of Lynch syndrome (LS) cases. The aim of this study was the identification of patients harboring constitutional MLH1 epimutations in a set of 34 patients with a clinical suspicion of LS, MLH1-methylated tumors and non-detected germline mutations in mismatch repair (MMR) genes. MLH1 promoter methylation was analyzed in lymphocyte DNA samples by MS-MLPA (Methylation-specific multiplex ligation-dependent probe amplification). Confirmation of MLH1 constitutional methylation was performed by MS-MCA (Methylation-specific melting curve analysis), bisulfite sequencing and pyrosequencing in different biological samples. Allelic expression was determined using heterozygous polymorphisms. Vertical transmission was evaluated by MS-MLPA and haplotype analyses. MS-MLPA analysis detected constitutional MLH1 methylation in 2 of the 34 individuals whose colorectal cancers showed MLH1 methylation (5.9%). These results were confirmed by bisulfite-based methods. Both epimutation carriers had developed metachronous early-onset LS tumors, with no family history of LS-associated cancers in their first-degree relatives. In one of the cases, the identified MLH1 constitutional methylation was monoallelic and results in MLH1 and EPM2AIP1 allele-specific transcriptional silencing. It was present in normal somatic tissues and absent in spermatozoa. The methylated MLH1 allele was maternally transmitted and methylation was reversed in a daughter who inherited the same allele. MLH1 methylation screening in lymphocyte DNA from patients with early-onset MLH1-methylated LS-associated tumors allows the identification of epimutation carriers. The present study adds further evidence to the emerging entity of soma-wide MLH1 epimutation and its heritability.

BackgroundWithin the ICD and DSM review processes there is growing debate on the future classification and status of adjustment disorders, even though evidence on this clinical entity is scant, particularly outside specialised care.AimsTo estimate the prevalence of adjustment disorders in primary care; to explore whether there are differences between primary care patients with adjustment disorders and those with other mental disorders; and to describe the recognition and treatment of adjustment disorders by general practitioners (GPs).MethodParticipants were drawn from a cross-sectional survey of a representative sample of 3815 patients from 77 primary healthcare centres in Catalonia. The prevalence of current adjustment disorders and subtypes were assessed face to face using the Structured Clinical Interview for DSM-IV Axis I Disorders (SCID-I). Multilevel logistic regressions were conducted to assess differences between adjustment disorders and other mental disorders. Recognition and treatment of adjustment disorders by GPs were assessed through a review of patients' computerised clinical histories.ResultsThe prevalence of adjustment disorders was 2.94%. Patients with adjustment disorders had higher mental quality-of-life scores than patients with major depressive disorder but lower than patients without mental disorder. Self-perceived stress was also higher in adjustment disorders compared with those with anxiety disorders and those without mental disorder. Recognition of adjustment disorders by GPs was low: only 2 of the 110 cases identified using the SCID-I were detected by the GP. Among those with adjustment disorders, 37% had at least one psychotropic prescription.ConclusionsAdjustment disorder shows a distinct profile as an intermediate category between no mental disorder and affective disorders (depression and anxiety disorders).

Objectives The objectives were to determine the levels of general practitioner (GP) recognition of anxiety disorders and examine associated factors. Methods An epidemiological survey was carried out in 77 primary care centers representative of Catalonia. A total of 3815 patients were assessed. Results GPs identified 185 of the 666 individuals diagnosed as meeting the Structured Clinical Interview for DSM-IV Axis I Disorders (SCID-I) criteria for any anxiety disorder (sensitivity 0.28). Regarding specific anxiety disorders, panic disorder was registered in just three of the patients who, according to the SCID-I, did not meet the criteria for this condition .Generalized anxiety disorder was recorded by the GP in 46 cases, 4 of them being concordant with the SCID-I (sensitivity 0.03). The presence of comorbid hypertension was associated with an increased probability of recognition. Emotional problems as the patients' main complaint and additional appointments with a mental health specialist were associated with both adequate and erroneous recognition. Being female, having more frequent appointments with the GP and having higher levels of self-perceived stress were related to false positives. As disability increased, the probability of being erroneously detected decreased. Conclusion GPs recognized anxiety disorders in some sufferers but still failed with respect to differentiating between anxiety disorder subtypes and disability assessment.

Childhood adversities (CAs) have consistently been associated with mental health problems in childhood and adulthood. However, few studies have employed appropriate statistical methods that take into account overlap among CAs, and many of the ones that did so were based on insufficiently complex models. The present paper studies the prevalence of a wide variety of CAs, as well as their relationship to the onset of mental disorders in a representative sample of a Spanish population.The study is part of the ESEMeD-Spain project, a cross-sectional household survey, which included a nationally representative sample of the Spanish adult population. CAs' associations with lifetime prevalence of mental disorders were estimated using discrete-time survival analysis with person-years as the unit of analysis.Of our sample, 20.6% reported at least one CA, of whom 24% reported more than one CA. Parental death, parental mental disorder, family violence, economic adversity, physical and sexual abuse were associated with different groups of mental disorders. CAs were associated with the onset of mental disorders during several stages of life. Simulations suggest that CAs were associated with 12.6% of all disorders, 10.8% of mood disorders, 5.8% of anxiety disorders, 27% of substance disorders and 29.7% of externalising disorders.Prevalences of CAs in the Spanish population are lower than those found in other high-income countries, especially when compared to the USA. In Spain, different CAs were associated with the onset of a number of mental disorders, although these associations were not as frequent as in other countries. Although lower than in other countries, the association between CAs and mental health in Spain should be considered relevant. Specific health policies and prevention programmes are needed in order to decrease this burden.

Trabajo presentado a la Alzheimer's Association 2011 International Conference on Alzheimer's Disease (ICAD 2011) celebrada en Paris del 16 al 21 de julio de 2011.

El pdf del artículo es la versión post-print., Background & Aims: Liver steatosis enhances ischemia/reperfusion (I/R) injury and is considered a primary factor in graft failure after liver transplantation. Although previous reports have shown a role for qualitative steatosis (macrovesicular vs. microvesicular) in hepatic I/R injury, no studies have compared side by side the specific contribution of individual lipids accumulating in fatty liver to I/R damage. Methods: We used nutritional and genetic models of micro and macrovesicular fatty livers exhibiting specific lipid profiles to assess their susceptibility to normothermic I/R injury. Results: Unlike choline-deficient (CD) diet-fed mice, characterized by predominant liver triglycerides/free fatty acids (TG/FFA) accumulation, mice fed a cholesterol-enriched (HC) diet, which exhibited enhanced hepatic cholesterol loading in mitochondria, were highly sensitive to I/R-induced liver injury. In vivo two-photon confocal imaging revealed enhanced mitochondrial depolarization and generation of reactive oxygen species following hepatic I/R in HC-fed but not in CD-fed mice, consistent with decreased mitochondrial GSH (mGSH) observed in HC-fed mice. Moreover, ob/ob mice, characterized by increased hepatic TG, FFA, and cholesterol levels, were as sensitive to I/R-mediated liver injury as mice fed the HC diet. Livers from ob/ob mice displayed increased StAR expression and mitochondrial cholesterol accumulation, resulting in mGSH depletion. Interestingly, atorvastatin therapy or squalene synthase inhibition in vivo attenuated StAR overexpression, mitochondrial cholesterol loading, and mGSH depletion, protecting ob/ob mice from I/R-mediated liver injury. Conclusions: Cholesterol accumulation, particularly in mitochondria, sensitizes to hepatic I/R injury, and thus represents a novel target to prevent the enhanced damage of steatotic livers to I/R-mediated damage. © 2010 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved., The work was supported by grants SAF2006-06780, SAF2008-02199, SAF2008-04974 and SAF2009-11417 (Plan Nacional de I+D), PI070193 and PI09/00056 (Institut de Salud Carlos III), by CIBEREHD from the Institut Carlos III, the Fundación Mutua Madrileña and the center grant P50-AA-11999 Research Center for Liver and Pancreatic Diseases, US National Institute on Alcohol Abuse and Alcoholism, USA.

The pathogenesis and treatment of nonalcoholic steatohepatitis (NASH) are not well established. Feeding a diet deficient in both methionine and choline (MCD) is one of the most common models of NASH, which is characterized by steatosis, mitochondrial dysfunction, hepatocellular injury, oxidative stress, inflammation, and fibrosis. However, the individual contribution of the lack of methionine and choline in liver steatosis, advanced pathology and impact on mitochondrial S-adenosyl-l-methionine (SAM) and glutathione (GSH), known regulators of disease progression, has not been specifically addressed. Here, we examined the regulation of mitochondrial SAM and GSH and signs of disease in mice fed a MCD, methionine-deficient (MD), or choline-deficient (CD) diet. The MD diet reproduced most of the deleterious effects of MCD feeding, including weight loss, hepatocellular injury, oxidative stress, inflammation, and fibrosis, whereas CD feeding was mainly responsible for steatosis, characterized by triglycerides and free fatty acids accumulation. These findings were preceded by MCD- or MD-mediated SAM and GSH depletion in mitochondria due to decreased mitochondrial membrane fluidity associated with a lower phosphatidylcholine/phosphatidylethanolamine ratio. MCD and MD but not CD feeding resulted in increased ceramide levels by acid sphingomyelinase. Moreover, GSH ethyl ester or SAM therapy restored mitochondrial GSH and ameliorated hepatocellular injury in mice fed a MCD or MD diet. Thus, the depletion of SAM and GSH in mitochondria is an early event in the MCD model of NASH, which is determined by the lack of methionine. Moreover, therapy using permeable GSH prodrugs may be of relevance in NASH.

et al., Alcohol is recognized as a direct hepatotoxin, but the precise molecular pathways that are important for the initiation and progression of alcohol-induced tissue injury are not completely understood. The current understanding of alcohol toxicity to organs suggests that alcohol initiates injury by generation of oxidative and nonoxidative ethanol metabolites and via translocation of gut-derived endotoxin. These processes lead to cellular injury and stimulation of the inflammatory responses mediated through a variety of molecules. With continuing alcohol abuse, the injury progresses through impairment of tissue regeneration and extracellular matrix (ECM) turnover, leading to fibrogenesis and cirrhosis. Several cell types are involved in this process, the predominant being stellate cells, macrophages, and parenchymal cells. In response to alcohol, growth factors and cytokines activate many signaling cascades that regulate fibrogenesis. This mini-review brings together research focusing on the underlying mechanisms of alcohol-mediated injury in a number of organs. It highlights the various processes and molecules that are likely involved in inflammation, immune modulation, susceptibility to infection, ECM turnover and fibrogenesis in the liver, pancreas, and lung triggered by alcohol abuse., The work described in this review was supported by National Health and Medical Research Council (NH&MRC) of Australia/Department of Veteran’s Affairs (DVA) (Seth); NIH, NIAAA grant R01AA013168 (D’Souza El-Guindy); NH&MRC and the Cancer Council of New South Wales, Australia (Apte); CIBEREHD and grant PI070193 (Instituto de Salud Carlos III, Spain), and by grant P50-AA-11999 (Research Center for Liver and Pancreatic Diseases, U.S. National Institute on Alcohol Abuse and Alcoholism) (Mari); BMBF (HepatoSys) and the European Alcohol Research Foundation (ERAB) (Dooley).

"La Fiesta de las Flores se celebra anualmente en la ciudad fronteriza de El Paso, Texas, bajo el patrocinio de lulac, una asociación de latinos de base ciudadana. Como organizadora de esta actividad, lulac entreteje redes sociales dentro de la comunidad hispanohablante, lo que ayuda a construir una identidad comunitaria y una memoria compartida. El certamen de belleza resalta el atractivo y los logros de las jóvenes de la comunidad latina. Los eventos de "la fiesta" están relacionados con asuntos políticos que atañen, de manera central, a la vida de los latinos. Esteproyecto involucra a un muy amplio sector de la sociedad, en el que las jóvenes participantes del certamen de belleza son un elemento clave. No sólo serán portadoras de una corona en un juego de simulación, sino que obtendrán recursos para seguir estudiando. Su participación en el concurso interiorizará el orgullo por su color, su lengua materna y la cultura del país de origen. Todo esto echando mano de las pautas y códigos sociales aceptados y vigentes en la sociedad de la que ahora forman parte."

The role of cholesterol in Alzheimer’s disease (AD) has been linked to the generation of toxic amyloid β peptides (Aβ). Using genetic mouse models of cholesterol loading, we examined whether mitochondrial cholesterol regulates Aβ neurotoxicity and AD pathology. Isolated mitochondria from brain or cortical neurons of transgenic mice overexpressing SREBP-2 (sterol regulatory element binding protein 2) or NPC1 (Niemann-Pick type C1) knock-out mice exhibited mitochondrial cholesterol accumulation, mitochondrial glutathione (mGSH) depletion and increased susceptibility to Aβ1– 42-induced oxidative stress and release of apoptogenic proteins. Similar findings were observed in pharmacologically GSH-restricted rat brain mitochondria, while selective mGSH depletion sensitized human neuronal and glial cell lines to Aβ1–42-mediated cell death. Intracerebroventricular human Aβ delivery colocalized with mitochondria resulting in oxidative stress, neuroinflammation and neuronal damage that were enhanced in Tg-SREBP-2 mice and prevented upon mGSH recovery by GSH ethyl ester coinfusion, with a similar protection observed by intraperitoneal administration of GSH ethyl ester. Finally, APP/PS1 (amyloid precursor protein/presenilin 1) mice, a transgenic AD mouse model, exhibited mitochondrial cholesterol loading and mGSH depletion. Thus, mitochondrial cholesterol accumulation emerges as a novel pathogenic factor in AD by modulating Aβ toxicity via mGSH regulation; strategies boosting the particular pool of mGSH may be of relevance to slow down disease progression., This work was supported in part by the Plan Nacional de Investigación Científica, Desarrollo e Innovación Tecnológica Grants SAF2005-03923 and SAF2006-06780, by Research Center for Liver and Pancreatic Diseases Grant P50 AA 11999 funded by the National Institute on Alcohol Abuse and Alcoholism, and by the Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas supported by the Instituto de Salud Carlos III. A.C. is supported by the Ramon y Cajal Research Program (Ministerio de Educación y Ciencias).

[Background]: Alcohol-induced liver injury is associated with decreased S-adenosyl-l-methionine (SAM)/S-adenosyl-l-homocysteine (SAH) ratio and mitochondrial glutathione (mGSH) depletion, which has been shown to sensitize hepatocytes to tumor necrosis factor (TNF)., [Aims]: As the effect of alcohol on mitochondrial SAM (mSAM) has been poorly characterized, our aim was to examine the status and transport of mSAM in relation to that of mGSH during alcohol intake., [Methods]: Sprague–Dawley rats were pair fed Lieber–DeCarli diets containing alcohol for 1 to 4 weeks and liver fractionated into cytosol and mitochondria to examine the mSAM transport and its sensitivity to membrane dynamics., [Results]: We found that cytosol SAM was depleted from the first week of alcohol feeding, with mSAM levels paralleling these changes. Cytosol SAH, however, increased during the first 3 weeks of alcohol intake, whereas its mitochondrial levels remained unchanged. mGSH depletion occurred by 3 to 4 weeks of alcohol intake due to cholesterol-mediated impaired transport from the cytosol. In contrast to this outcome, the transport of SAM into hepatic mitochondria was unaffected by alcohol intake and resistant to cholesterol-mediated perturbations in membrane dynamics; furthermore cytosolic SAH accumulation in primary hepatocytes by SAH hydrolase inhibition reproduced the mSAM depletion by alcohol due to the competition of SAH with SAM for mitochondrial transport. However, alcohol feeding did not potentiate the sensitivity to inhibition by SAH accumulation., [Conclusions]: Alcohol-induced mSAM depletion precedes that of mGSH and occurs independently of alcohol-mediated perturbations in membrane dynamics, disproving an inherent defect in the mSAM transport by alcohol. These findings suggest that the early mSAM depletion may contribute to the alterations of mitochondrial membrane dynamics and the subsequent mGSH down-regulation induced by alcohol feeding., The work presented was supported in part by the Research Center for Liver and Pancreatic Diseases, P50 AA 11999 funded by the U.S. National Institute on Alcohol Abuse and Alcoholism; Plan Nacional de I + D grants SAF (2005-03923, 2005-03943, 2006-06780) and CIBERehd supported by Instituto de Salud Carlos III, Spain. Anna Colell is a Ramón y Cajal investigator.

et al., The aim of this study was to demonstrate the existence of alterations in glutathione and cholesterol homeostasis in brain mitochondria from alcoholic rats. Glutathione concentration decreased, whereas oxidized glutathione and cholesterol contents increased in these organelles, suggesting the ethanol-induced generation of reactive oxygen species, and the impairment of mitochondrial uptake of glutathione, possibly due to the increase in cholesterol deposition. The release of apoptogenic proteins was increased after stimulating mitochondria from the brain of alcoholic rats with atractyloside. As a conclusion, chronic alcohol consumption might sensitize brain mitochondria to apoptotic stimuli, and promote the subsequent release of apoptotic proteins., This work was supported in part by Funds from Ministerio de Ciencia e Innovación (SAF 2007- 66801), Plan Nacional de Investigación Científica, Desarrollo e Innovación Tecnológica (I+D+I), Ingenio 2010 initiative, RTA (Red de Transtornos Adictivos) and Consolider programms from Instituto de Salud Carlos III, Coopernicus-Santander Programm of the Universidad CEU- Cardenal Herrera, Dirección General de Drogodependencias (Generalitat Valenciana) and FEPAD (Fundación para el Estudio, Prevención y Ayuda a la Drogadicción).

To estimate the comorbidity of mental disorders with chronic physical conditions and to assess their independent and combined effects on health-related quality of life (HRQOL). Face-to-face cross-sectional survey of adult attendants to public primary care (PC) centres from Catalonia (Spain). A total of 3,815 out of 5,402 selected patients provided data for this study. We report frequency of chronic physical conditions among participants with mental disorders and the contribution of each mental disorder and chronic physical condition to HRQOL. Chronic pain is the most frequent condition among those with mental disorders (74.54%). The effect of chronic physical conditions on HRQOL is rather minor when compared to the effect of mental disorders (especially mood disorders). However, chronic pain plays an important role in HRQOL loss. Mood disorders and chronic pain negatively affect HRQOL of PC patients. Especial efforts should be made to detect and treat mental disorders and chronic pain at this level.

Currently in Spain, 4 depot antipsychotics are available: flufenazine, pipotiazine, zuclopentixol and risperidone. The objectives of the present study are: a) to evaluate the efficacy of depot vs. oral forms of typical and atypical antipsychotics available in Spain for treating patients with schizophrenia; b) to compare the efficacy of different depot antipsychotics; c) to evaluate cost-effectiveness of typical and atypical depot and oral antipsychotics.Systematic review of the literature between January 1980 and March 2007. Pharmaceutical companies of depot preparations were contacted aiming to include unpublished material.A total of 15 studies were included (13 journal manuscripts and 2 posters provided by the industry). Concordance between evaluators was moderate-high. The quality of selected studies was moderate-low. There were no differences in the efficacy between depot and oral risperidone. Efficacy of depot risperidone was higher than oral olanzapine (there were no differences regarding tolerability) and higher and better tolerated than oral zuclopentixol. The evidence was controversial when comparing the efficacy of depot and oral flufenazine. There were no differences when comparing the efficacy between depot flufenazine and oral pimozide. Depot zuclopentixol was more efficient than the oral preparation for treating patients with schizophrenia and violent behaviour. Finally, there were no differences regarding the efficacy and tolerability between depot pipotiazine and depot flufenazine and between depot clopentixol and depot flufenazine.There is few high-quality scientific evidence comparing depot and oral antipsychotics or different depot antipsychotics available in Spain. Selected evidence does not allow to conclude that depot antipsychotics are more effective and better tolerated than oral ones.

Comunicacion presentada en la Alzheimer's Association International Conference on Alzheimer's Disease, celebrada del 26 al 31 de julio de 2008 en Chicago (Estados Unidos)

[Background & Aims]: Nuclear factor κB (NF-κB) is the master regulator of tumor necrosis factor (TNF) susceptibility. Although mitochondrial glutathione (mGSH) depletion was shown to sensitize hepatocytes to TNF despite NF-κB activation, the mechanisms involved, particularly the role of Bax oligomerization and mitochondrial outer membrane (MOM) permeabilization, 2 critical steps in cell death, remained unexplored., [Methods]: TNF signaling at the premitochondrial and mitochondrial levels was analyzed in primary mouse hepatocytes with or without mGSH depletion., [Results]: Unexpectedly, we observed that TNF activates caspase-8 independently of NF-κB inactivation, causing Bid cleavage and mitochondrial Bax oligomerization. However, their predicted consequences on MOM permeabilization, cytochrome c release, caspase-3 activation, and hepatocellular death occurred only on mGSH depletion. These events were preceded by stimulated mitochondrial reactive oxygen species that predominantly oxidized cardiolipin, changes not observed in acidic sphingomyelinase (ASMase)−/− hepatocytes. Oxidized cardiolipin potentiated oligomerized Bax-induced MOM-like liposome permeabilization by restructuring the lipid bilayer, without effect on membrane Bax insertion or oligomerization. ASMase−/− mice with mGSH depletion by cholesterol loading were resistant to TNF-induced liver injury in vivo., [Conclusions]: Thus, MOM-localized oligomeric Bax is not sufficient for TNF-induced MOM permeabilization and cell death requiring mGSH-controlled ASMase-mediated mitochondrial membrane remodeling by oxidized cardiolipin generation., Supported in part by the Research Center for Liver and Pancreatic Diseases (P50 AA 11999) funded by the US National Institute on Alcohol Abuse and Alcoholism; Plan Nacional de ID grants SAF (2005-03923, 2005-03943, 2006-06780) and BFU2005-06095; CIBERehd (Centro de Investigación Biomédica en Red en el Área temática de Enfermedades Hepáticas y Digestivas) and FIS (04/1039, 06/0395, and 07/0193) grants supported by the Instituto de Salud Carlos III, Spain. M.M. is an IDIBAPS (Institut d’Investigacions Biomèdiques August Pi i Sunyer) Investigator. A.C. and A.M. are Ramon y Cajal Investigators. O.T. is a recipient of a predoctoral fellowship from the Basque Government.

TThe pathogenesis of alcohol-induced liver disease (ALD) is still poorly understood. One of the clues to its progression relates to the alcohol-mediated susceptibility of hepatocytes to cell death by reactive oxygen species (ROS) and inflammatory cytokines. Tumor necrosis factor alpha (TNF) has been considered a key ALD mediator with acidic sphingomyelinase (ASMase)-mediated ceramide generation playing a critical role. TNF receptor 1 and 2 knock-out mice or ASMase(-/-) mice exhibit resistance to alcohol-mediated fatty liver and cell death. Furthermore, alcohol feeding has been shown to sensitize hepatocytes to TNF due to the limitation of mitochondrial glutathione (mGSH) through impaired import of GSH from the cytosol due to altered membrane order parameter caused by mitochondrial cholesterol increase. Selective pharmacological depletion of mGSH sensitizes hepatocytes to TNF-mediated cell death, which reproduces the observations found with alcohol feeding. TNF signaling analyses in hepatocytes with or without mGSH depletion indicate that mGSH prevents cardiolipin peroxidation (CLOOH) formation by TNF-induced ROS via ASMase and that CLOOH cooperates with oligomerized Bax to cause mitochondrial outer membrane permeabilization through destabilization of the lipid bilayer via increased bilayer-to-inverted hexagonal phase transitions. Thus, activation of ASMase and cholesterol-mediated mGSH depletion both collaborate to promote alcohol-induced TNF-mediated hepatocellular damage, suggesting novel therapeutic opportunities in ALD., The work presented was supported in part by the Research Center for Liver and Pancreatic Diseases, P50 AA 11999; and grant 1R21 AA014135-01, funded by the US National Institute on Alcohol Abuse and Alcoholism; Plan Nacional de I + D grants SAF (2005-03923; 2005-03943; 2006-06780); and CIBERehd supported by Instituto de Salud Carlos III, Spain.

Suicide is a public health problem and it is increasing in Spain. The objective of this study is to analyze the prevalence and risk factors of suicide related outcomes (ideation, plan and attempt) using data from the ESEMeD-Spain project.This is a face-to-face household survey carried out in a probability representative sample of the adult general population of Spain. 5,473 subjects were interviewed using the Composite International Diagnostic Interview (CIDI 3.0), developed by the World Mental Health Survey Initiative.Lifetime prevalence of suicide ideation and attempts was 4.4% and 1.5%, respectively. Risk of suicide related outcomes was significantly higher among women (odds ratio [OR] = 2.3-2.7), younger cohorts (OR = 21.3-86), and lower education levels (OR = 5.3-6.4). Having a mental disorder was associated to an increased risk in all diagnostic categories, but especially in major depressive episode (OR = 5.3-6.8). Risk of suicide attempt was higher during the first year since the onset of ideation (OR = 30.2), decreasing thereafter.The prevalence of suicide related outcomes is low when compared with other countries. Results identified groups with higher risk (women, young, subjects with a mental disorder, psychiatric comorbidity and recent suicidal ideation) in which suicide prevention could show benefits.

P0307 MELATONIN-INDUCED APOPTOSIS OF HepG2 CELLS IS ENHANCED BY AUTOPHAGY SUPPRESSION R. Ordonez, A. Fernandez, L. Martinez, S. Nunez, S. CarbajoPescador, A. Baulies, N. Prieto-Dominguez, C. Garcia-Ruiz, J.C. Fernandez-Checa, J.L. Mauriz, J. Gonzalez-Gallego. Institute of Biomedicine (IBIOMED), University of Leon (Spain), Leon, Centro de Investigacion Biomedica en Red: Enfermedades hepaticas y Digestivas (CIBERehd). Instituto de Salud Carlos III, Madrid, Department of Cell Death and Proliferation, Instituto Investigaciones Biomedicas de Barcelona, CSIC, Barcelona, and Liver Unit-Hospital Clinic-IDIBAPS, Barcelona, Spain E-mail: rordf@unileon.es

Comunicacion presentada en la Alzheimer's Association 10th International Conference on Alzheimer's Disease and Related Disorders, celebrada del 15 al 20 de julio de 2006 en Madrid (Espana)

The etiology of progression from steatosis to steatohepatitis (SH) remains unknown. Using nutritional and genetic models of hepatic steatosis, we show that free cholesterol (FC) loading, but not free fatty acids or triglycerides, sensitizes to TNF- and Fas-induced SH. FC distribution in endoplasmic reticulum (ER) and plasma membrane did not cause ER stress or alter TNF signaling. Rather, mitochondrial FC loading accounted for the hepatocellular sensitivity to TNF due to mitochondrial glutathione (mGSH) depletion. Selective mGSH depletion in primary hepatocytes recapitulated the susceptibility to TNF and Fas seen in FC-loaded hepatocytes; its repletion rescued FC-loaded livers from TNF-mediated SH. Moreover, hepatocytes from mice lacking NPC1, a late endosomal cholesterol trafficking protein, or from obese ob/ob mice, exhibited mitochondrial FC accumulation, mGSH depletion, and susceptibility to TNF. Thus, we propose a critical role for mitochondrial FC loading in precipitating SH, by sensitizing hepatocytes to TNF and Fas through mGSH depletion. © 2006 Elsevier Inc. All rights reserved., The work presented was supported in part by the Research Center for Liver and Pancreatic Diseases, P50 AA 11999; and grant 1R21 AA014135-01, funded by the US National Institute on Alcohol Abuse and Alcoholism; Plan Nacional de I+D grants SAF (2002-3564, 2003-04974, 2005-03923, 2005-03943, and 2006-0678); and Red Temática de Investigación Cooperativa G03/015, Red de Centros C03/02 PI 05/1285 and FIS (04/1039, 03/0426, 02/3057 and 02/0339) grants supported by Instituto de Salud Carlos III, Spain

The potential of mesophilic anaerobic digestion for the treatment of fats of different origin through co-digestion with the organic fraction of municipal solid wastes (OFMSW) has been evaluated. Co-digestion process was conducted in a pilot plant working in semi-continuous regime in the mesophilic range (37 °C) and the hydraulic retention time (HRT) was 17 days. During the start-up period the digester was fed with increasing quantities of a simulated OFMSW (diluted dry pet food). When the designed organic loading was reached, co-digestion process was initiated. The fat used consisted of waste from the food industry (animal fat), its composition was very similar to that of the simulated OFMSW in relation to the long-chain fatty acid (LCFA) profile. Fat content in the feedstock was gradually increased up to 28%, and hence the organic loading, with stable operation of the digester. Animal fat was suddenly substituted by vegetable fat maintaining the organic loading. No accumulation of LCFA or volatile fatty acids (VFA) was detected in either case. After a short adaptation period, total fat removal throughout the experiment was over 88%, whereas biogas and methane yields were very similar to those of simulated OFMSW. In conclusion, anaerobic co-digestion of OFMSW and fat wastes appears to be a suitable technology to treat such wastes, obtaining a renewable source of energy from biogas.

2 ) tests. ACox regression model was used to evaluate the Relapse Free Survival (RFS) and the Colon Cancer specific Survival. Results: MSI status could be determined by genotyping in 350 tumor samples. 48 patients (14%) had MSI high (MSI-H) tumors and 302 (86%) had microsatellite stable (MSS) tumor. BRAF status could be determined in 380 tumor samples. 58 cases (15%) were BRAF mutated tumors and 322 (85%) were BRAF wild type tumors. Tumor-stroma ratio was analyzed in 407 cases. 176 tumors (43%) had more than 50% intra-tumor stroma and were classified as stroma-high. MSI-H tumors were significantly located in the right colon (X 2 p-value = 1.03e-5), were poorly differentiated (X 2 p-value = 0.003) and had more than 12 lymph nodes resected (X 2 p-value = 0.037). MSI-H tumors were associated with BRAF mutation (X 2 p-value = 0.02) and stroma-low (X 2 p-value = 0.0005). When a molecular prognostic risk classification was performed, patients with MSI-H and stroma-low suggested a low risk of relapse comparing to MSI-H/stroma-high, MSS/stroma-low and MSS/ stroma-high (Hazard Ratio = 3.15; 95% CI, 0.76 - 13.1, p-value = 0.0602). Conclusion: MSI-H tumors are typically poorly differentiated, located in the right colon and have BRAF mutation as well as low intra-tumor stroma. Our results suggest that low-stroma could partly explain prognosis in patients with MSI-H stage II tumors.

3629 Background: Different clinico-pathological factors are used to define a group of patients with high-risk stage II colon cancer (CC) that may benefit of adjuvant treatment. Moreover, several molecular markers, such as microsatellite instability (MSI) and BRAF, have been widely investigated as prognostic factors. Recently a high stroma component (EMT+ subtypes) has also been associated with poor outcome. The aim of the study is to analyze the prognostic value of MSI, BRAF and tumor-stroma ratio in a prospective series of stage II CC patients. Methods: FFPE tissue from 432 stage II CC patients operated at Hospital Universitari de Bellvitge (1996 - 2006) were included in the study. MSI status was assessed by the analysis of 5 mononucleotide repeat markers (BAT-25, BAT-26, NR-21, NR-24 and MONO-27). BRAF V600E mutation was analyzed by single strand conformation polymorphism technique. Tumor-stroma ratio was analyzed by immunohistochemistry. Associations between molecular factors and clinical features were assessed by Chi-Squared (X2) tests. A Cox regression model was used to evaluate the Relapse Free (RFS) and the Colon Cancer specific Survival. Results: MSI status could be determined in 350 patients. 48 (14%) had MSI high (MSI-H) tumor. BRAF status could be assessed in 380 cases. 58 (15%) cases were BRAF mutated. Tumor-stroma ratio was analyzed in 407 tumor samples. 176 (43%) tumors had more than 50% intra-tumor stroma and were classified as stroma-high. MSI-H tumors were significantly located in right colon (X2 p-value = 1.03e-5), were poorly differentiated (X2 p-value = 0.003) and had more than 12 lymph nodes resected (X2 p-value = 0.037). MSI-H tumors were associated with BRAF mutation (X2 p-value = 0.02) and stroma-low (X2 p-value = 0.0005). When a molecular prognostic risk classification was performed, patients with MSI-H /stroma-low suggested a low risk of relapse comparing to MSI-H/stroma-high, microsatellite stable (MSS)/stroma-low and MSS/stroma-high patients (HR = 3.15; 95 CI, 0.76 – 13.1, p-value = 0.0602). Conclusions: MSI-H tumors have BRAF mutation as well as low intra-tumor stroma. Our results suggest that tumor-stroma ratio can explain differential prognosis in MSI-H stage II tumors.