BMC Medicine research article, Kivity et al. demonstrated novel work showed that the 16/6 Id antibody impaired visual memory and spatial memory by causing hippocampal injury in mice. Given differences in the cross-reactivity of each autoantibody with the nervous system, the clinical features might be different and diverse in NPLE. Identification of autoantibody targets could lead to the development of novel therapies. Investigators and clinicians should consider not only the inhibition of autoantibody synthesis but also the protection of neuronal cells in the treatment strategy for NPLE.Systemic lupus erythematosus (SLE) is a multi-system inflammatory disorder characterized by the presence of several autoantibodies, including anti-double-stranded DNA. Neuropsychiatric (NP)LE contributes to the prognosis of SLE, and is divided into 19 NPLE syndromes. Its mechanisms are mediated through autoantibodies, complement components, and cytokines. The pathophysiology and diagnosis of NPLE are diverse and complicated. Recent studies have shown that several autoantibodies cross-react with human brain tissue and cause NPLE symptoms in SLE. It is known that in mice, depression and hippocampus-related memory impairment are induced by anti-ribosomal P antibody and anti-NR2 antibody, respectively. In a http://www.biomedcentral.com/1741-7015/11/90See related Research article:  Systemic lupus erythematosus (SLE) is a multi-system inflammatory disorder characterized by the presence of autoantibodies directed against double-stranded (ds) DNA. The prevalence ranges from 20 to 150 cases per 100,000 population, and it seems to be increasing, partly because the disease is recognized more readily and partly because of longer survival [NPLE is classified by the American College of Rheumatology (ACR) into 19 neuropsychiatric syndromes . The difN-methyl-D-aspartate (NMDA) receptor subunit 2 (NR2) in SLE [Recent novel studies have identified some aspects of the pathophysiology of NPLE. Some anti-dsDNA antibodies cross-react with the ) in SLE . NMDA re) in SLE ,5.Mycobacterium tuberculosis[The 16/6 idiotype (Id) antibody, which was the focus of a recent study by Kivity et al. [erculosis. Depositerculosis, and leverculosis. These fTo understand if the 16/6 Id IgG antibody is able to induce neurological effects, Kivity et al. compared the cognitive and behavioral performance of C3H female mice that had been injected with the human 16/6 Id IgG antibody (16/6 Id mice subset) and those injected with a commercial human IgG (control mice subset) [In addition, the brain pathology of these mice was examined to establish the potential mechanism by which the 16/6 Id IgG antibody is able to exert its neurological effects. In the brain tissue, increased microglial activation was seen in the hippocampus and amygdala, but not in the neurocortex or piriform cortex. In addition, the number of astrocytes in the hippocampus was found to be increased. Taken together, these results show that in mice, the 16/6 Id antibody causes impairment of both visual and spatial memory through hippocampal injury, and might selectively cross-react with some antigens in the hippocampus.Kivity et al. also showed that, in addition to the anti-ribosomal P and anti-NR2 antibodies, another autoantibody, anti-16/6 Id antibody, can cross-react with human brain tissue and cause NPLE symptoms in SLE . Therefo2+ influx [Therapy of NPLE is currently difficult. Although corticosteroids and immunosuppressive agents, such as cyclophosphamide, are broadly effective for NPLE, the condition is occasionally refractory to these treatments. Moreover, brain-tissue-reactive autoantibodies might cause irreversible neuronal degeneration via apoptosis. For instance, anti-ribosomal P antibody targets a neuronal surface protein, causing calcium influx and apoptosis . These a+ influx . Kivity + influx . The aut+ influx .It is plausible, therefore, that the wide variety of NPLE syndromes might be caused by differences in the recognition of brain tissue by lupus autoantibodies, such as anti-ribosomal P, anti-NR2, and anti-16/6 Id antibodies. Identification and evaluation of such differences would perhaps be useful in developing therapies for NPLE.2+, or regulate the deposition of autoantibodies in NPLE. Memantine is a drug used to treat Alzheimer’s disease, which modulates intracellular Ca2+ by blocking NMDA receptors. In addition, the DWEYS pentapeptide, which the anti-NR2 antibody recognizes as an antigen, prevents the anti-NR2 antibody from being deposited in tissues and mediating neuronal excitotoxicity in mice [In the future, it is hoped that new agents will be developed to improve the prognosis for NPLE. The efficacy of such new agents should be determined through their ability to protect neuronal cells, modulate intracellular Ca in mice .As Kivity et al. have shown, several autoantibodies against brain tissue are involved in NPLE. Given the differences in the cross-reactivity of each autoantibody with the nervous system, this may explain the difference and diversity of the clinical features in NPLE. Investigators and clinicians should consider not only inhibition of autoantibody synthesis but also protection of neuronal cells when investigating treatment strategies for NPLE.N-methyl-D-aspartate; NPLE: Neuropsychiatric lupus erythematosus; PNS: Peripheral nervous system; SLE: Systemic lupus erythematosusACR: American college of rheumatology; BBB: Blood–brain barrier; CNS: Central nervous system; ds: Double-stranded; NMDA: The authors declared that they have no competing interest.TG drafted the manuscript, and YK and HY read and revised it. All authors have given final approval of the final manuscript to be published.TG is an assistant professor in the Institute of Rheumatology (IOR), Tokyo Women’s Medical University (TWMU), and is interested in the neurological complications associated with connective tissue disease. YK is an associate professor of Medicine and Rheumatology in TWMU. HY is a professor of Medicine and Rheumatology, and a director of the IOR, TWMU. All authors are board-certified members of the Japan College of Rheumatology.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1741-7015/11/91/prepub


The epithelial-to-mesenchymal transition (EMT) is a cellular dedifferentiation process that is critical to development, wound healing and metastasis. Like other cell state transitions, such as differentiation, EMT is accompanied by genome-wide epigenetic reprogramming. However, the relationship between reprogramming and functional changes in the cell is poorly understood. In an A549 non-small cell lung cancer EMT model system we observed changes in chromatin state between epithelial and mesenchymal states. Multivariate analyses were applied to paired  ChIP-seq data for 18 histone modifications/variants and expression microarray data. We observed epigenetic co-regulation of genes associated with EMT, as well as their proximal enhancers. We also observed epigenetic activation or repression of functionally distinct sets of enhancers. These genes and enhancers are regulated and bound by a small set of transcription factors, specifically AP-1, NF-κB and c-Myc. These transcription factors themselves also a show an epigenetic profile similar to the EMT-related genes. Together, these observations suggest a chromatin-mediated transcriptional feedback mechanism that establishes and maintains the phenotypic switch.


Postprandial lipemia (PPL) is likely a risk factor for cardiovascular disease but these changes have not been well described and characterized in a large cohort. We assessed acute changes in the size and concentration of total and subclasses of LDL, HDL, and VLDL particles in response to a high-fat meal. Participants (n = 1048) from the Genetics of Lipid-Lowering Drugs and Diet Network (GOLDN) Study who ingested a high-fat meal were included in this analysis. Lipids were measured at 0 hr (fasting), 3.5 hr, and 6 hr after a standardized fat meal. Particle size distributions were determined using nuclear magnetic resonance spectroscopy. Analyses were stratified by baseline triglycerides  and gender. The effect of PPL on changes in lipoprotein subclasses was assessed using repeated measures ANOVA.Postprandially, LDL-C, HDL-C, VLDL-C, and triglycerides increased regardless of baseline triglyceride status, with the largest increases in VLDL-C and TG; however, those with elevated triglycerides demonstrated larger magnitude of response. Total LDL particle number decreased over the 6-hour time interval, mostly from a decrease in the number of small LDL particles. Similarly, total VLDL particle number decreased due to reductions in medium and small VLDL particles. Large VLDL particles and chylomicrons demonstrated the largest increase in concentration. HDL particles demonstrated minimal overall changes in total particle number.We have characterized the changes in LDL and VLDL particle number, and their subclass patterns following a high-fat meal. Most studies use the measurement of fasting lipoproteins to assess cardiovascular disease (CVD) risk ,2; howevThus, we sought to describe in a large cohort, acute changes in lipoprotein  particle sizes and numbers following a high-fat meal and to determine if the lipoprotein particle changes would vary by baseline triglyceride concentrations and gender.Table Total LDL-C levels demonstrated decreases between 0 and 6 hours following the high-fat meal . Further, the magnitude of the VLDL-C levels was higher among men than women and among hyper-triglyceridemic compared to normo-triglyceridemic individuals Figure . Total tHyper-triglyceridemic participants had higher baseline concentrations of all LDL particle subclasses compared to normo-triglyceridemic participants Tables  and 3. AIn response to a high-fat meal, normo- and hyper-triglyceridemic men displayed similar patterns of change in HDL particle size concentration distributions; however, the magnitudes of these patterns differed in hyper-compared to normo-triglyceridemic participants. In men, there was no effect on large HDL particle concentrations, but significant increases in the concentration of medium HDL particles  as well as significant decreases in small HDL particle concentrations . Furthermore, total HDL particle number significantly increased among men in response to the high-fat meal (p < 0.0001), which can be attributed to the large increases in medium HDL particle size concentrations.Among both normo- and hyper-triglyceridemic individuals, women responded differently to the high-fat meal than men. Normo-triglyceridemic women demonstrated a significant decrease in total HDL particle number (p < 0.0001), while there was no overall change among hyper-triglyceridemic women in response to the meal (33 to 32 mmol/L in normo-triglyceridemic and 35 mmol/L in hyper-triglyceridemic). Additionally, these subgroups demonstrated different responses for large HDL particle concentrations where normo-triglyceridemic women displayed significant increases (8 to 9 mmol/L), and hyper-triglyceridemic women displayed significant decreases in large HDL particle concentration . For both normo- and hyper-triglyceridemic women, medium and small HDL particle concentrations demonstrated similar responses to the high-fat meal: medium HDL particle concentrations had significant increases , and small HDL particle concentrations had significant decreases . Similar to LDL size, no substantial change in HDL size was demonstrated in response to the high-fat meal despite its statistical significance (p < 0.0001).Results of the PPL VLDL particle size subclass analysis were similar in pattern for men and women, but the magnitude of change and concentration values were different, with women, on average, having lower concentrations. Total VLDL particle number decreased significantly among both normo- and hyper-triglyceridemics . Large VLDL particle concentrations increased significantly . Medium and small VLDL particle concentrations demonstrated significant decreases (p < 0.0001 for both men and women). Finally, VLDL particle size significantly increased (p < 0.0001) among normo- and hyper-triglyceridemic men and women.The present study focused on the PPL response to an acute (1-time) high-fat meal using data derived from the Genetics of Lipid-Lowering Drugs and Diet Network (GOLDN) Study. We sought to describe lipoprotein particle subclass number, size, and concentrations stratified by gender and baseline triglyceride status. Although the pattern of change observed for both men and women was qualitatively similar, the magnitude of these changes appeared to be more exaggerated in men compared to women, especially for VLDL and TG response. Similar to the response demonstrated by gender, the pattern of change was qualitatively similar for normo- and hyper-triglyceridemic individuals; however, the magnitude was more exaggerated among hyper-triglyceridemic than normo-triglyceridemic individuals.Specifically, in terms of lipoprotein particles for both normo- or hyper-triglyceridemic individuals, we observed little change in total particle number over the 3 time points  for either total LDL or HDL particle number. In contrast, for normo-triglyceridemic subjects, total VLDL particle number appeared to rise above baseline at 3.5 hours and drop considerably below baseline at 6 hours after the high-fat meal. This pattern was considerably different in hyper-triglyceridemic individuals where, in both men and women, we observed a precipitous drop relative to baseline which continued from the 3.5 to the 6 hour time point.Despite expected baseline differences between normo- and hyper-triglyceridemic individuals, there was a reduction in small LDL particle concentrations secondary to the PPL challenge. In normo-triglyceridemic individuals there was a persistent decline from baseline, while in hyper-triglyceridemic men and women small LDL particle concentration appeared to plateau.The major pattern we observed among HDL particle subclasses was a significant shift to more medium HDL particles and fewer small HDL particles with a non-significant increase in HDL-C (p > 0.05 for men and women). Decewicz et al  observedIn our study, the total number of VLDL particles decreased during the postprandial period which was due to an increase in large VLDL and chylomicrons along with more substantial decreases in medium and small VLDL subclasses. A decrease in medium VLDL and total VLDL particles was also observed among the diabetic subjects in the study by Colhoun et al . Our stuDuring the PPL response to a high-fat meal, many changes occur during lipoprotein metabolism. It has been shown that HDL becomes enriched with TG, leading to increased removal of HDL from the plasma, thereby decreasing HDL-C . Also thThis study differs from the majority of previous studies examining dietary influences on PPL that were conducted in smaller study populations, utilized different dietary interventions, and did not report changes in LDL particle subclasses ,12. For The strengths of this study are its large sample size, the use of a standardized acute fat load, and the use of NMR technology to characterize particle size subclasses. To our knowledge, it is the largest PPL study conducted to date. Our study did not measure serum apolipoprotein B and is not a CVD outcome or mechanistic study; therefore, we do not know for certain how or why these lipoprotein particle distributional shifts occur or how they might impact CVD risk. Nonetheless, among hyper-triglyceridemic individuals, the prolonged return to baseline levels may add to an already elevated atherogenic profile .Additionally, we did not obtain information on the activity level of enzymes and proteins that are known to be activated during the PPL response, such as CETP and HL . SpecifiPostprandial lipemia persists for at least 6 hours following a high-fat meal. We confirmed that men differ from women in the PPL response, and that the PPL response varies based on baseline triglyceride profile. We demonstrated an increase in IDL particles, especially in normo-triglyceridemic women, and an increase in triglycerides in both men and women. Large HDL also changed in hyper-triglyceridemic women (17% decrease was observed), which is interesting in view of the clinically observed greater risk of CVD in hyper-triglyceridemic women compared to men . OverallThe Genetics of Lipid-Lowering Drugs and Diet Network (GOLDN) study population consisted of 189 families who were recruited from 3-generational pedigrees from the NHLBI Family Heart Study Minneapolis, MN, and Salt Lake City, UT, field centers. The overarching intent of GOLDN was to identify loci contributing to phenotypic variation in response to a high-fat meal and fenofibrate therapy. The Institutional Review Boards at the University of Alabama, the University of Minnesota, the University of Utah, and Tufts University approved the study protocol.Participants had to meet the following criteria: White , male or female, ≥ 18 years of age, fasting triglycerides (TGs) < 1500 mg/dL, willingness to participate in the study and attend the scheduled clinic exams, part of a family with at least 2 members in a sibship, AST and ALT tests within normal range, and creatinine ≤ 2.0. Exclusion criteria for the fenofibrate intervention included the following: history of liver, kidney, pancreas, or gall bladder disease or malabsorption; pregnancy or women of childbearing potential not using an acceptable form of contraception; insulin use; known hypersensitivity to fenofibrate; pancreatitis within 12 months prior to enrollment; and current use of warfarin and/or nutraceuticals. Subjects using hypolipidemic drugs were asked to consult with their physicians to determine whether they could be taken off medications for 4 weeks prior to and for the duration of the study; informed consent for drug withdrawal was obtained. Only subjects who remained untreated with any prescription or over-the-counter hypolipidemic medication were eligible.Participants underwent 2 interventions: an acute fat loading dietary challenge and a short-term (3 weeks) open-label intervention trial of fenofibrate. The specific methodology of GOLDN is reported elsewhere ,24. For 2 of body surface area (2.93 MJ/m2 body surface area). The meal composition was 83% of calories from fat, 14% from carbohydrates, and 3% from protein. The meal was formulated to have a cholesterol content of 240 mg and a polyunsaturated:saturated fat ratio of 0.06. Based on these guidelines, the average individual ingested 175 mL of heavy whipping cream (39.5% fat) combined with 7.5 mL powdered, instant, non-fat, dry milk, and blended with ice. To increase palatability of the drink, 15 mL of chocolate- or strawberry-flavored syrup was also added. Participants had 15 minutes to ingest this meal and were required to fast a minimum of 8 hours prior to the meal. Immediately before ingestion, we drew blood samples on all participants , and then again at 3.5 and 6 hours after the high-fat meal.The high-fat challenge followed the protocol of Patsch et al . The calSerum and EDTA-anticoagulant tubes were collected and processed using a standardized protocol, aliquoted, and stored at -70°C until time of use. Analysis was completed on all stored samples at the end of the study and all samples for an individual were processed in the same batch to reduce measurement error. Triglyceride measurements were calculated using the glycerol-blanked enzymatic method on the Roche COBAS FARA centrifugal analyzer (Roche Diagnostics Corporation). Hyper-triglyceridemia was defined as a baseline triglyceride ≥ 150 mg/dL. HDL-C was calculated using the same procedure as TG measurement after precipitation of non-HDL cholesterol with magnesium/dextran. LDL-C measurement employed a homogeneous direct method  on a Hitachi 911 Automatic Analyzer.Proton nuclear magnetic resonance (NMR) spectroscopy was used to determine LDL, HDL, and VLDL particle size subclass concentrations . Measured amplitudes after exposure to a 400 MHz magnet and known distinct lipid methyl group NMR spectroscopic profiles were used to determine the subclass concentrations ,26-30. C2 and Fisher exact tests. To compare crude means, we used ANOVA and Student's t-test. Results are expressed as least squares mean (standard error). We examined changes in subclass particle size concentrations that occurred during the dietary fat challenge  using repeated-measures ANOVA in which the family was modeled as a random variable. All statistical calculations were performed using SAS 9.1 .All analyses were conducted separately for men and women and stratified by baseline hyper-versus normo-triglyceride levels (TG > 150 vs TG ≤ 150) due to significant interactions between gender and baseline triglyceride concentrations in analyses for LDL, HDL, and VLDL. To determine the significance of differences in percentages between male and female normo- and hyper-triglyceridemic individuals, we used the Pearson cALT: alanine transaminase; AST: aspartate transaminase; CETP: cholesteryl ester transfer protein; CVD: cardiovascular disease; GOLDN: Genetics of Lipid-Lowering Drugs and Diet Network Study; HDL-C: high-density lipoprotein cholesterol; HL: hepatic lipase; IDL-C: intermediate-density lipoprotein; LDL-C: low-density lipoprotein cholesterol; NMR: nuclear magnetic resonance (spectroscopy); PPL: post-prandial lipemia; TG: triglyceride; VLDL-C: very low-density lipoprotein cholesterolThe authors declare that they have no competing interests.MKW conducted the statistical analyses and drafted the manuscript. AO and EKK provided critical input within their respective areas of expertise during analysis, drafting, review, and revision. SPG, PNH, MYT, RJS, and JMO were critical participants in study design and provided input within their respective areas of expertise during analysis, drafting, review, and revision. DKA was the primary designer of the study and oversaw and contributed to all stages of analysis and authoring. All authors read and approved the final manuscript.


In a large sample (n = 399) of healthy adults aged from 18 to 85 years, we analyzed age-dependent differences in resting-state (RS) (task-independent) functional connectivity (FC) of a set of brain regions derived from a previous fMRI study. In that study, these regions had shown an age-related loss of activation specificity in visual-attention  or sensorimotor  tasks. In addition to these dedifferentiated regions, the FC analysis of the present study included “task-general” regions associated with both attention and sensorimotor systems  as defined via meta-analytical co-activation mapping. Within this network, we observed both selective increases and decreases in RS-FC with age. In line with regional activation changes reported previously, we found diminished anti-correlated FC for inter-system connections . Our analysis also revealed reduced FC between system-specific and task-general regions, which might reflect age-related deficits in top-down control possibly leading to dedifferentiation of task-specific brain activity. Together, our results underpin the notion that RS-FC changes concur with regional activity changes in the healthy aging brain, presumably contributing jointly to age-related behavioral changes.Healthy aging is accompanied by structural and functional changes in the brain, among which a loss of neural specificity  is one of the most consistent findings. Little is known, however, about changes in interregional integration underlying a dedifferentiation across  Effective information processing depends on the integrity of communication between the different nodes of functional brain systems. Through normal aging, substantial changes within and between brain networks occur  study, we investigated functional activation patterns in the sensorimotor and the visual attention systems in a large sample of healthy subjects between 20 and 70 years of age , FC between regions of the visual-attention or sensorimotor systems  that show a dedifferentiated recruitment pattern with increasing age. In this context it should be noted that temporally correlated brain activity in spatially distinct regions  may not only arise from direct interaction . In the second step, age-related changes in task-independent FC for this set of seed regions were analyzed in the same sample. This approach of examining the RS-FC including task-general regions in a large sample of healthy subjects should provide insights into age-related changes in the functional coupling between brain regions involved in sensorimotor and visual attention processing.In the present study, we investigated changes of RS-FC between functional seeds that showed an age-related dedifferentiated recruitment pattern and task-general regions that are functionally related to them. The functional seeds were defined based on results from a previous fMRI study  to assess the co-activation pattern of each seed  and converted to Z-scores for visualization  and the MACM-derived task-general seed regions conjointly represented the combined set of our seed regions, comprising sensorimotor area OP4 and attention-related dPMC and area 7A, as fMRI-based seed regions, as well as SMAr and AIC as task-general regions Figure .An RS-FC network analysis was implemented to analyze the task-independent FC between these seed regions and its change with age for all possible connections between the combined set of seed regions .Mean = 41.8, SD = 16.8, Median = 41, IQR = 29). All participants  were without any record of neurological or psychiatric disorders and gave their written informed consent to participate in the study. The data were contributed by four sites . The EPI images were first corrected for head movement by affine registration using a two-pass procedure. For normalization the mean EPI images were segmented into gray matter, white matter and cerebrospinal fluid using the “unified segmentation” approach  contrast . Subsequently, the same Fisher-Z transformed correlation coefficients for each connection were rank-correlated with age to test for age-related changes in inter-regional coupling. The results of this correlation analysis were regarded significant if they passed a threshold of p < 0.05, corrected for multiple comparisons using the false discovery rate. Finally, we analyzed the connectivity in the 100 youngest and 100 oldest participants for all connections showing significant age-related changes, to corroborate the correlational findings. Moreover, this analysis permits a more detailed analysis of each age-related change of RS-FC to test, e.g., if an age-related FC increase represents a significant change from negative to positive FC or from negative to absent FC. The former pattern would imply an inversion of the RS communication , while the latter would imply a loss of communication between brain regions.FC analyses may be influenced by several confounds such as physiological processes, e.g., fluctuations related to cardiac and respiratory cycles and in particular motion-related effects  SMAr with bilateral OP4 and bilateral AIC; (ii) left AIC with bilateral OP4; (iii) right AIC with bilateral OP4 and right area 7A; (iv) bilateral dPMC each with ipsilateral area 7A; (v) right DPMC with left area 7A; (vi) interhemispheric connections between bilateral regions AIC, dPMC, OP4, and area 7A  was found for the following connections: (i) SMAr with bilateral area 7A and right dPMC; (ii) bilateral OP4 to bilateral area 7A and dPMC; (iii) right AIC with bilateral dPMC and left AIC with left dPMC , some connections showed corresponding FC, whereas other connections showed no FC congruency . Positive RS-FC was predominantly observed between brain regions associated with similar functional systems Figure , whereasIn turn, we found that negative FC was predominantly found between regions from different domains. These results underline an intrinsic, functionally driven organization of the human brain and are supported by studies showing that regions with apparently opposing functionality are negatively correlated in their RS-FC . Nevertheless, we also found a few connections showing an increase in FC. The healthy aging brain is thus not only subject to functional decline but rather responds selectively to presumable structural or biochemical neuronal changes in the elderly. Again, this finding is particularly important in that it underlines that connectivity is not reduced per se, which may potentially be deemed to reflect systematic confounds. Rather, we found decreased (closer to zero), increased, or unchanged connections in our large sample.First of all, it is noteworthy that not all connections showed age-related changes, arguing against a general and unspecific age-related decline in the task-independent functional coupling between brain regions. Instead, it appears that changes across the lifespan occur selectively. Connections that showed age-related changes, however, mostly featured reductions of functional correlations, as both positive and negative RS-FC moved closer to zero with age. In other words, the task-independent functional correlation  within and between the present networks appears to be diminished in older adults  may indicate more task specific impairments for older adults. In line with this, the interaction of the visual-attention and sensorimotor systems seems to be deteriorated in older adults , as found in our former study  adults often recruit more frontal regions to successfully perform a visual attention task , functional MRI, and neurochemical measurements, may extend the understanding of the causalities on FC changes in the aging brain. Second, since the analyzed functional networks reflect only parts of the visual-attention or sensorimotor system that showed an age-related loss of neural specificity, inferences from our results on age-related RS-FC changes between entire functional networks have to be regarded with caution. Nevertheless, our study indicates that healthy aging is associated with task-independent connectivity changes within and across task-specific brain network nodes. Furthermore, the current sample was derived by four different sites. Although all participants were screened for psychiatric and neurological disorders, the threshold for sub-clinical cognitive impairments may vary between them. Hence, the sample may contain some patients with sub-clinical symptoms. Finally, in the current sample we were unable to relate individual FC parameters to performance. Further studies should hence combine performance measurements with neuroimaging during task performance and task-free states in the same participants. This would permit investigating more direct relations of different age-related neural changes, their interdependencies, and their association with performance (see, e.g., Andrews-Hanna et al., The present study corroborates the notion that RS activity reflects the functional organization of the human brain (De Luca et al., The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Personality and self-rated health have been linked previously to cognitive outcome in late life. However, these associations have not been shown among the oldest old. This study examined relationships between personality, self-rated health, and cognitive function in a selected sample of Ashkenazi Jewish centenarians  aged 95 to 106 who lived independently in the community. Personality was measured using the Personality Outlook Profile Scale (POPS), a brief measure that was validated in this population. Self-rated health was assessed by participants’ subjective rating of their present health, and Mini-Mental Status Examination was used to determine cognitive function. Results showed positive associations of the Positive Attitude Towards Life domain of the POPS and self-rated health with participants’ current cognitive function. These associations remained significant even after adjusting for the effects of participants'age, gender, marital status, education, and history of medical illnesses. Further exploratory analysis using structural equations modeling showed significant associations among the three variables, but demonstrated a borderline significant level of mediating effect of personality on the relationship between self-rated health and cognition. These results reemphasized the independent roles of personality and self-rated health on centenarians’ cognitive outcomes. Future studies will further elucidate the impact of personality and self-rated health on cognitive outcomes in the oldest old. Centenarians have been suggested to be an example of successful aging due to their delay or escape of major illnesses . Many ceAmong the contributors to successful aging and longevity, personality has been particularly associated with favorable health outcomes, longevity/mortality, and cognitive function in late life -10. WhilThe Five Factor Model proposes that five universal traits that explain personality variability including neuroticism/emotional stability, extraversion, conscientiousness, openness to experience, and agreeableness . SeveralAnother determinant that affects cognitive function in late life is self-rated health . In thisThe current study investigated the role of personality and self-rated health in cognitive function through assessing cross-sectional relationships among personality, self-rated health, and cognitive function in a group of Ashkenazi Jewish centenarians who lived independently in the community with less than a part-time home/nursing care. This group was thought to be a model of successfully aging. Personality was assessed by the Personality Outlook Profile Scale (POPS) , a briefDemographic, personality, behavioral, and clinical characteristics of the sample (N = 68) are presented in Table The results of the multiple regression analyses Table  showed tWe conducted further exploratory analyses to test a mediation model in which PATL mediates the relationship between self-rated health and MMSE. This model was tested to clarify the relationships among these variables. Personality was thought to partially mediate the relationship between self-rated health and cognition as maladaptive personality may lead to distorted perceptions about one's health status . Prior tThe present study investigated the cross-sectional relationshipsof centenarians’ personality andhealth-rated health to cognitive function in a genetically and environmentally homogenous group of Ashkenazi Jewish centenarians. We demonstrated small but significant associationsof the POPS-PATL and self-rated healthwith centenarians’ cognitive function. These associations remained significant even after adjusting for the effects of age, gender, marital status, education, and history of medical illnesses, suggesting that these were small yet meaningful effects. Further exploratory analyses to test a mediation model showed a borderline significant indirect effect of PATL on the relationship between self-rated health and MMSE.Findings of the present study are generally consistent with previous studies which support the relationships of personality and self-rated health to cognitive function in the general older adult populations , 31-36. In summary, the purpose of this study was to examine cross-sectional associations of personality and self-rated health to cognitive functioningin Ashkenazi Jewish centenarians who were living independently in the community. This study adds to a growing body of knowledge which suggests that personality as well as self-rated health may play important roles in cognitive function in advanced age. Although personality was also associated with self-rated health, the mechanisms which explain these relationships among personality, self-rated health, and cognition, which may also have therapeutic implications for prevention and/or treatment for cognitive decline, remain unclear. Thus, further investigation of long-term effects of personality and self-rated health on cognitive functioning is warranted.241 Ashkenazi Jewish centenarians  were recruited to the study. Based on participants’ living arrangements and levels of homecare, 166 participants residing in a skilled-nursing home or living in the community with full-time homecare were excluded from the study. 75 participants living independently in the community or in senior housing without full-time homecare and/or skilled medical/nursing care were included in the study. These participants were selected since they may represent a model for successfully aging. Of those 75, 7 participants were further excluded from the analyses due to their missing responses. Thus, 68 participants (ages 95-106y) were included in the analyses of this study. Details of the study's sampling and recruitment have been described previously , 48. AshDetailed study procedures have been described previously , 48. A sThe structured 98-item questionnaire, developed by the Longevity Genes Project at Albert Einstein College of Medicine, measures participants’ personal demographics, personality characteristics, health/medical history, self-rated health, and health-related behaviors , 50. ParThis is a brief self- and informant-report measure of personality which represents personality phenotypes which may contribute to successful aging. POPS consists of two domains and six items: Positive Attitude Towards Life  and Emotional Expression (EE: expressing emotions openly and not bottling up emotions). The Centenarian Questionnaire served as the basis for the development of this measure. Items were developed using a rational content-based approach and derived from extant longevity research and theoretical concepts. They reflected centenarians’ lifetime personality and related behaviors. Two robust domains consisting of six items, PATL and EE, were selected from original eleven items. A composite score of each domain was creased based on a mean z-score of respective items for each domain. Higher scores reflect more favorable personality characteristics. In our previous study, these domains showed adequate internal consistency (α =.65 for PATL and α =.63 for EE), and this measure was cross-validated with the NEO-Five Factor Inventory and Life Orientation-Test in self- and informant-report groups .This is a brief measure of cognitive status in adults and older adults. The MMSE is a 30-item measure which tests an individual's orientation, attention, calculation, recall, language, and motor skills at a given point in time. It is used to screen for cognitive impairment, estimate the severity of cognitive impairment, and follow the course of cognitive changes over time. A cutoff score of 24 has been recommended for the detection of dementia . For cenAnalyses were completed with the statistical software, SPSS Statistics (Version 17.0). Multiple regression analyses were used to evaluate the relationships of the POPS to self-rated health and MMSE scores as well as the relationship between self-rated health and MMSE scores. The hierarchical regression analyses were conducted to adjust for the effects of demographic variables including age, gender, marital status, and years of education and history of medical illnesses on MMSE scores. In an exploratory analysis using Stata Version 12.1, structural equations modeling was used to determine the mediating role of PATL on the relationship between self-rated health and cognitive function.


Improvements in the techniques for metabolomics analyses and growing interest in metabolomic approaches are resulting in the generation of increasing numbers of metabolomic profiles. Platforms are required for profile management, as a function of experimental design, and for metabolite identification, to facilitate the mining of the corresponding data. Various databases have been created, including organism-specific knowledgebases and analytical technique-specific spectral databases. However, there is currently no platform meeting the requirements for both profile management and metabolite identification for nuclear magnetic resonance (NMR) experiments.1H-NMR metabolomic profiles, is designed (i) to provide a knowledgebase of curated plant profiles and metabolites obtained by NMR, together with the corresponding experimental and analytical metadata, (ii) for queries and visualization of the data, (iii) to discriminate between profiles with spectrum visualization tools and statistical analysis, (iv) to facilitate compound identification. It contains lists of plant metabolites and unknown compounds, with information about experimental conditions, the factors studied and metabolite concentrations for several plant species, compiled from more than one thousand annotated NMR profiles for various organs or tissues.MeRy-B, the first platform for plant http://www.cbib.u-bordeaux2.fr/MERYB/index.php.MeRy-B manages all the data generated by NMR-based plant metabolomics experiments, from description of the biological source to identification of the metabolites and determinations of their concentrations. It is the first database allowing the display and overlay of NMR metabolomic profiles selected through queries on data or metadata. MeRy-B is available from  The set of low-molecular weight  molecules of an organism, organ or tissue is referred to as the metabolome . Metabol1H-NMR) [The chemical diversity and complexity of the plant metabolome constitutes a real challenge, even for a given species, because the diversity of metabolites and their concentration ranges remains huge. It is therefore impossible to profile all metabolite families  simultaneously through a single extraction and with only one analytical technique. Most metabolomics projects therefore use several analytical strategies in parallel ,20. Seve1H-NMR) ,22.1H-NMR and GC-MS have been applied to polar extracts for the study of primary metabolites. 1H-NMR technology has been widely used as a high-throughput technique for non targeted fingerprinting with little or no sample preparation [1H-NMR and is ideal for the detection of volatile metabolites, but high-boiling point metabolites require two-step derivatization [paration ,24. It hparation , despiteparation . GC-MS itization .1H-NMR is that the profiles obtained are often comparable, even between different instruments or different field magnitudes [The relative quantification of a hundred hydrophilic metabolites can be achieved, but comparisons of sets of GC-MS metabolomics profiles obtained in different laboratories remain difficult. For the study of secondary metabolites, LC-MS analysis is generally the method of choice. Extracts are injected directly, without derivatization. LC-MS is generally used for metabolomic profiling  with relgnitudes ,32, prov1H-NMR, generate large amounts of raw, processed and analyzed data, which must be well managed if they are to generate useful knowledge. Various web-based software platforms are available for managing and making use of metabolomics data. These software platforms include metabolite spectral databases, such as the Golm Metabolome Database (GMD) and the Human Metabolome DataBase (HMDB). The GMD [http://www.metabolome.jp/software/knapsack-database or Phenolexplorer [http://www.phenol-explorer.eu dedicated to the polyphenol content of food. MetaboAnalyst [1H NMR spectral signatures (after data reduction with bucketing) for urine samples for example. MetaboAnalyst does not handle NMR spectra but only processed data (peak list or buckets list) in tabular csv files. Each of these applications is useful, but none constitutes a complete tool for managing, analyzing and sharing plant NMR metabolomics data.Metabolomics facilities, including those using  The GMD  provides The GMD ,34 is an The GMD  are also The GMD  and Setu The GMD , two web The GMD , provideexplorer , a biblioAnalyst  is an oni) a spectral database combined with ii) a knowledgebase for plants, iii) an easy-to-use metabolomic spectral visualization tool and iv) a metabolomic data analysis tool. Taking these requirements into account, we have developed a plant metabolomics platform (with public or private access) for the storage, management, visualization, analysis, annotation and query of NMR fingerprints or quantitative profiles and quantified metabolite. This platform has been named MeRy-B, for Metabolomics Repository Bordeaux. MeRy-B facilitates profile discrimination through the visualization of spectral data by either modular spectrum overlay  or multivariate statistical analysis. It can also construct a knowledgebase of plant metabolites determined by NMR, including metabolite concentration data when available, with minimal information about experimental conditions in the context of scientific publications, and can be used for the re-analysis of combined experiments. Furthermore, MeRy-B provides tools for the identification of metabolites by comparisons of spectra for plant extracts with spectra available in the MeRy-B knowledgebase.Given the types of metabolomics resources available (listed in ), and thData storage and database building tools are required for the storage and analysis of present and future metabolomics data. MeRy-B therefore takes into account the recommendations of initiatives concerning the extent and types of metadata (information associated with the data or data about the data) to be stored for each metabolomics experiment: MiAMET ,43, StanThe architecture of MeRy-B Figure  is basedAdditions to the database are made principally through web interfaces, with various forms. These data input forms are accessible to registered users. Other metadata are uploaded, stored in files and made available for consultation. For example, all protocols are collected in PDF format files, as such files are already available as part of the quality assurance approach operating in most laboratories: standard operating procedures (SOPs) are available and users therefore waste little time uploading these data into the MeRy-B database.The database is structured according to the steps in a metabolomics experiment and therefore consists of four principal components: "Experimental design" Figure  "Analytihttp://www.ncbi.nlm.nih.gov/Taxonomy/, Plant Ontology Consortium http://www.plantontology.org/ and Environment Ontology http://environmentontology.org/. A Project is defined as an entity comprising a set of experiments carried out on one species by a laboratory, at a particular geographic site. Within a given Project, each Experiment is carried out within a particular set of environmental conditions, such as 'control' or 'stress'. A protocol file in PDF format is uploaded for each step in the experiment: growth, harvest and storage of the biological samples. Five types of biological factor potentially contributing to definition of the experimental design are defined: organ or tissue, genotype, genetic background, developmental stage and environmental conditions.The Experimental Design component describes the biological source and protocols for plant growth, sample harvest, extract preparation and storage Figure . The expi.e. 1H-NMR).MeRy-B also manages metadata concerning the analytical part of the experiments. The preparation of analytical samples , parameters of analytical instruments and spectrum processing metadata are described in PDF protocols Figure . The PDFAnalytical Profiles to assist the user with the input of repetitive analytical metadata. An Analytical Profile consists of an instrument description, an extraction method description and the various types of protocol: extraction, analytical and processing.MeRy-B can generate Spectral data component describes spectrum format and processed data  must be converted into JCAMP-DX format . Spectra may be uploaded as data that have already been preprocessed by commercial software . Alternatively, MeRy-B provides custom-designed signal processing methods for 1r NMR data. These methods include noise suppression, baseline correction . Each spectrum, whether modeled or not, is linked to an Experimental Design and an Analytical Profile.The a Figure . MeRy-B ransform ), deconvransform ) and theCompounds component provides information about the identification of a given compound and its quantification, when available . A specific nomenclature is used to allocate identifiers to the unknown compounds, to link these unknown signatures in the various spectra of the database. For example, an interesting singlet peak was detected on a spectrum at 1.9 ppm. This unknown compound is thus named unkS1.90: with S for singlet and 1.90 for the chemical shift expressed in ppm in agreement with the recommendations of MSI [The e Figure . Each spcompound/), or as s of MSI . A putatAdministration component  if access via the public user account is set up by the project's owner.The user responsible for creating a project automatically becomes its "owner". The owner of a project can provide temporary or permanent access rights  to other users on his or her project. By default, a 1H NMR spectrum visualization (the self-signed certificate is available on the"About MeRy-B" page). The backend statistical computing and visualization operations are carried out with functions from the R packages and Perl scripts. Data storage, treatment and querying have been developed with Perl, XML and web services technologies, such as SOAP.MeRy-B is a PostgreSQL relational database accessible through a web interface developed in the PHP language. The web interface is rendered dynamic by the use of JavaScript and AJAX technologies. The application is maintained on a Linux server. A Java applet has been developed for MeRy-B fulfills two needs. First, each registered user, as a project owner, creates projects and deposits his or her own data and associated metadata into the application for storage, consultation, visualization and analysis. At this point, there is no curation team deciding whether or not an upload should be allowed. However, the administrator is alerted when a project is rendered public and he verifies this new inclusion of data. Second, all users are allowed to search the MeRy-B knowledgebase constructed from the information provided by all previous project owners (public data), for the re-analysis and comparison of data sets and to facilitate compound identification. The utility of MeRy-B for each of these cases is detailed below. A user manual illustrated with screenshots is available from the MeRy-B website for a more detailed description.Data capture module in the MeRy-B database: (1) experimental design, (2) analytical metadata, (3) spectral data, and (4) compounds (lists and/or quantifications). Three main steps were used 1) creation of the users account and project, 2) population of the database with the user's data, and 3) analysis and visualization of the user's data. The aim of the tomato study was to characterize differences between the metabolic profiles of two interdependent tissues, seeds and flesh, from the same fruits, during fruit development, by means of a metabolomics approach. Before the creation of the MeRy-B project, it was necessary to define an informative title and to decide which factors should be taken into account for subsequent data visualization and analysis. Two factors, tissue (Seed vs Flesh) and developmental stage, were clearly identified and guided the coding of the biological samples and the organization of the data in the database. Two experiments were created: Tomato-Seed and Tomato-Flesh.Data uploading and consultation are illustrated here, as a use case, with the data and metadata of a published study on tomato . Four maGrowth, Harvest and Storage as pdf files through the Protocols menu: PG- Tomato - Metabolomics - 2006, PH- Tomato - Metabolomics - 2006 and PS-Tomato-UMR619-1. The 'Environmental Condition', 'Study Type' and 'Tissue/Organ' were selected from drop-down lists: Normal, Growth chamber study and Seed or Fruit. Several controlled vocabularies were also required, such as Culture Localization, Genotype Lycopersicum esculentum var 'Ailsa Craig'. These requests were sent to the MeRy-B administrator who created and added this new controlled vocabulary. The five Developmental stages were then created by the user for each experiment: from FF.01 fruit size 30% (8 days post anthesis or DPA) to FR.04 fruit ripening complete (45 DPA) and the genotype was chosen (Ailsa Craig). The Analytical Metadata component was then created and documented with a description of the NMR spectrometer (in Instrument Menu), NMR sample preparation (conditions of sample preparation by resuspension or reconstitution in solvent (in the Methods menu)), the protocols used for extraction/preparation of the samples (PE-Tomato - Metabolomics -2006), NMR acquisition (PA- Tomato - Metabolomics -2006) and NMR processing (PP- Tomato - Metabolomics -2006). The next step was the creation of Analytical Profiles. Sample coding was described in the extraction protocol: e.g. Sx.y.z indicates Seed sample at x days post anthesis, y indicates the pool or biological replicate number and z, the technological replicate. During the transformation of NMR spectra from Bruker format to JCAMP-DX format, the spectra were renamed with the above code. They were then imported into MeRy-B through the Spectral Data module.Once the user's account had been created by the MeRy-B administrator, an accession number was allocated: T06002 . The project was created by uploading the three protocols describing Data consultation menu, the overlay module was particularly useful for checking the quality of spectra and the View module for checking the consistency of biological replicates. In addition, as spectra are colored according to criteria chosen by the user, such as by experiment, developmental stage or sample code, visual inspection and identification of the spectral areas specific to a tissue . These tools included standardization of bucket intensities followed by principal component analysis (PCA) or analysis of variance (ANOVA) Figures  and 2d, This MeRy-B output for the T06002 tomato project was consistent with the findings of the previous study , which hCompound, and then Add compound. The list of identified and/or quantified metabolites established was downloaded via 'Download the quantifiable compounds list' and opened with spreadsheet software on a PC (e.g. MS Excel) for completion with the quantification data from each NMR spectrum. This file was then uploaded into MeRy-B. The quantitative data can be visualized for the entire T06002 project through the menu Data consultation, Projects, Compounds , the experimental data and related metadata can be consulted through the Data consultation module and its various interfaces, providing either a global view or a detailed view. The complete experimental design, by project, is available through the Project Details function, which provides an overview on a single web page  are shared with the metabolomics community. Thus, MeRy-B can be used as a knowledgebase. Three helpful tools allow the sorting, visualization and export of the data already stored into the database: the search Spectral Data and search Compound under the tab labeled Search spectral data" tool can be used to visualize a MeRy-B spectrum in a matrix of interest  from a species of interest or a related species. A multicriterion search of metadata results in direct display of the corresponding spectra. For example, 190 spectra of tomato (Lycopersicon esculentum) pericarp obtained on a 500 MHz Bruker Avance at pH 6 in D2O solvent were available for public consultation on March 2011. In addition, users can obtain the peak list for each spectrum, the corresponding identified or unidentified compounds and their concentrations. The graphical view of each spectrum is interactive, making it possible to zoom in and focus on a region of the spectrum, to overlay the spectrum and to observe detected peaks. Figures containing NMR spectra in publications are often very small and not interactive. This tool is of particular interest for "beginners" with no experience with a particular tissue or plant matrix. In addition, there are often few published data dealing with the composition of the plant tissue, organ or biofluid and literature searches are time-consuming. MeRy-B currently compiles data for hundred metabolites in four species and eight tissues or organs, together with the corresponding metadata.The "Search compound" tool enables users to carry out searches of previously detected compounds stored in the MeRy-B knowledgebase. Three types of search may be carried out: (i) a compound search , (ii) a chemical shift search for 1H-NMR data  after the selection of the 1H NMR technique and (iii) advanced searches corresponding to a combination of both these types of search. For example, a new user observes a singlet at 9.08 ppm in tomato at pH 6. He or she then tries to identify this compound by looking for identified compounds described in the MeRy-B knowledgebase as a singlet close to 9.08 ppm ± 0.2. The search returns one compound: trigonelline, with an external link to the KEGG compound card. The user can then check whether the other three chemical shifts of trigonelline were also detected on his/her NMR spectrum. In addition, another link provides all the information available about each compound in MeRy-B via a "MeRy-B card" (MBC) . This card also highlights quantitative differences between species, tissues, organs or experiments for a given compound. One or several "MeRy-B cards" are returned for each chemical shift and/or compound search. Comparisons must take into account the possible use of different quantification units. Units are always provided on MeRy-B cards to prevent inappropriate comparisons.The ") Figure . ChemicaQuery Builder is a useful tool for queries and for the export of -omics data. We may need to add to the statistical treatments currently included in MeRy-B, nonlinear unsupervised multivariate methods, such as those based on neural networks, or supervised methods, such as the partial least square (PLS) method, included in tools such as Multi Experiment Viewer http://www.tm4.org/mev/ or MetaboAnalyst [http://www.cbib.u-bordeaux2.fr/x2dbi/. An example of the use of this module is provided in the Additional file Finally, oAnalyst , or otheoAnalyst , we deveA number of other databases worldwide are conceptually related to that presented here. However, MeRy-B has several advantages for plant metabolomics and for data management and analysis. MeRy-B is a single tool meeting the needs of the research community in this domain: one or several spectral databases, a knowledgebase for plants with an experimental design description, compound quantification files (when available) and search tools, several tools for spectrum visualization and statistics and one or several metabolite identification tools. These needs were previously met by using a series of databases and applications. Furthermore, MeRy-B was designed to improve the reporting of metabolomics research, based on MIBBI requirements: the MSI. Specialized ontological terms are used where applicable, for experimental design and analytical metadata for NMR, for example. Furthermore, MeRy-B can be used in three main ways: consultation within a project, consultation between projects and consultation of all the data present in the knowledgebase. When compared to human metabolite-oriented HMDB, MeRy-B is metabolomic profiles-oriented and dedicated to plants. When compared to the MetaboAnalyst web tool that handles processed data (peak lists or bucket lists), MeRy-B handles NMR spectra from visualization to statistical analysis using the corresponding metadata.Data consultation menu, with the Spectra Overlay module. Spectra are displayed in color according to the criteria chosen by the user, facilitating the visual inspection and identification of spectral regions varying as a function of the level of a given factor. This ready-to-use tool is much more powerful than the 'dual function' proposed by the manufacturers of NMR software, which is based exclusively on sample code. To our knowledge, this is the only spectrum visualization tool with this overlay feature available.One key feature of MeRy-B is the In publications, NMR metabolomic profiles are generally reduced to one or two representative spectra. These spectra are not interactive and their resolution is often too low for the reader to extract all the information they contain. In this context, MeRy-B is of particular interest for newcomers with no experience with a particular tissue or plant matrix, because it provides access to detailed experimental and analytical protocols, together with the composition of the corresponding plant sample. Such composition data are scarce in publications and their provision by MeRy-B is therefore of great potential utility. As in the HMDB database, the precise tissue or organ distribution of a compound within a plant, together with its quantification, constitute crucial information for MeRy-B users. Indeed, the level of quantification varies as a function of the tissue, organ or species of interest, and users can compare the amounts of a given compound between situations for the identification of potential biomarkers.13C NMR. The objective is to gather datasets generated by different analytical techniques, making it possible to benefit from their complementarity, as shown by recent publications [In the near future, we plan to make it possible to import and export experiment description data with the emerging ISA-tab format , which wications ,57. We aMeRy-B is a web-based application and database for the management and analysis of NMR plant metabolomics profiles, filling the gap in centralized information in this area. This platform manages all the data produced by a metabolomics experiment, from biological source description to compound identification. It also helps the user to analyze and to understand the data, by providing a number of visualization tools, for the visualization of NMR data by spectra overlay or multivariate statistical analyses, for example. By creating integrated visualizations, MeRy-B can provide biological insight. Furthermore, it provides information about metabolite quantification, making it possible to make comparisons between developmental stages, tissues, or environmental conditions. In March 2011, 20 users had a MeRy-B account, and 12 projects, 962 spectra and 100 compounds were available for public consultation in MeRy-B . All these data, cleverly exploited with MeRy-B tools, provide a useful knowledgebase for the sharing of plant NMR profiles and information relating to metabolites. This knowledgebase facilitates the identification of metabolites through comparisons between the spectra obtained for plant extracts and those present in the MeRy-B knowledgebase.Project name: MeRy-Bhttp://www.cbib.u-bordeaux2.fr/MERYB/home/home.phpProject home page: Browser requirement: the application is optimized for Firefox. However, it also works satisfactorily with Microsoft Internet Explorer version 7 and Safari.The user's web browser should support JAVA, to make it possible to benefit fully from MeRy-B.Users can create an account by submitting a form on the MeRy-B website. The user may populate the database him or herself, or assistance can be provided (see link on the website). MeRy-B is free to all academic users for data submission and their visualization and analysis.2O: deuterium oxide; DSS: 4,4-dimethyl-4-silapentane-1-sulfonic acid sodium salt; JCAMP-DX: the Joint Committee on Atomic and Molecular Physical data - Data Exchange format; KEGG: Kyoto Encyclopedia of Genes and Genomes. KEGG COMPOUND Database: http://www.genome.jp/kegg/compound/; MS: mass spectrometry; NMR: nuclear magnetic resonance; ppm: parts per million; SOAP: Simple Object Access Protocol; XML: Extensible Markup Language; TSP: (trimethylsilyl)propionic-2,2,3,3-d4 acid sodium salt;DADD and AM initiated the project. HFD, DJ and LG designed the DB. LG and DJ designed the web interface and implemented the DB and associated tools and developed the source code of the web application. CD actively populated the DB, tested the application and tools and provided feedback. HFD, LG, CD prepared the manuscript. DJ and SB participated in the drafting of the manuscript and its figures. CD provided studies for use cases. AM, CD and MN contributed to the critical reading of the manuscript. AM, DR, ADD and MN served as project advisors. All authors have read and approved the final submitted version.One example of use of Query Builder module in MeRy-B. This workflow tutorial with step-by-step and with screenshots illustrates how to reach the objective of extracting the list of the metabolites identified in the 1H-NMR spectra of project T06002: name, chemical shifts, groups and multiplicity.Click here for file


Repeated drug exposure can lead to an approach-bias, i.e. the relatively automatically triggered tendencies to approach rather that avoid drug-related stimuli. Our main aim was to study this approach-bias in heavy cannabis users with the newly developed cannabis Approach Avoidance Task (cannabis-AAT) and to investigate the predictive relationship between an approach-bias for cannabis-related materials and levels of cannabis use, craving, and the course of cannabis use.Cross-sectional assessment and six-month follow-up in 32 heavy cannabis users and 39 non-using controls.Approach and avoidance action-tendencies towards cannabis and neutral images were assessed with the cannabis AAT. During the AAT, participants pulled or pushed a joystick in response to image orientation. To generate additional sense of approach or avoidance, pulling the joystick increased picture size while pushing decreased it. Craving was measured pre- and post-test with the multi-factorial Marijuana Craving Questionnaire (MCQ). Cannabis use frequencies and levels of dependence were measured at baseline and after a six-month follow-up.Heavy cannabis users demonstrated an approach-bias for cannabis images, as compared to controls. The approach-bias predicted changes in cannabis use at six-month follow-up. The pre-test MCQ emotionality and expectancy factor were associated negatively with the approach-bias. No effects were found on levels of cannabis dependence.Heavy cannabis users with a strong approach-bias for cannabis are more likely to increase their cannabis use. This approach-bias could be used as a predictor of the course of cannabis use to identify individuals at risk from increasing cannabis use. Cannabis is the most commonly used illegal drug in most countries and treatment demands have increased strongly over the last decades –3. GrowiInability to control drug use is considered a core aspect of drug dependence –8. DespiCognitive biases for drug-related cues are thought to be behavioural manifestations of a drug-orientated motivational system. An abundance of studies, focusing mainly on alcohol and tobacco use, highlight roles of attentional, approach and evaluative biases in the development and maintenance of addictive behaviours  approach responses to cannabis cues in heavy cannabis users compared to non-users . The aimThirty-two heavy cannabis users and 41 controls aged 18–25 were recruited through advertisements on the internet and in cannabis outlets (coffee-shops). Groups were matched for age, gender and estimated intelligence ,34 Tabl. Heavy cCannabis use and related problems were assessed with the Cannabis Use Disorder Identification Test (CUDIT ) and a sAn adapted AAT ,29,30 waTest-sessions took place during the late afternoon and at the beginning of the evening. All participants were asked to refrain from alcohol and drug use 24 hours prior to testing. Each session started and ended with filling out the MCQ. After completing other questionnaires, administering cannabis use history and the diagnostic interview, participants performed the AAT. Six months later participants were contacted for a telephone interview on present drug use and related problems.t-tests were used to test if bias scores deviated significantly from zero within each group. Pearson's correlations and sequential multiple regression analyses were used to investigate associations between AAT bias scores, craving, measures of cannabis use and related problems at baseline and 6-month follow-up and tobacco smoking. To control for general biases in approach and avoid action-tendencies, partial correlations and regression analysis were conducted with neutral bias score as covariate.To correct for outliers, RTs below 200 ms, above 2000 ms and more than 3 standard deviations (SD) above and below the mean were removed for each participant. Error trials were removed. The bias score was calculated by subtracting median approach RT from median avoid RT for each image category. The subtraction resulted in a bias score for cannabis images and neutral images for each participant. As in previous AAT publications, median RTs were used because they are less sensitive to outliers than means ,29,30,44t71 = 1.30, P = 0.20), gender , IQ  and alcohol use , but there were more tobacco smokers among heavy cannabis users . In heavy cannabis users, post-test craving was higher for the MCQ purposefulness factor  (see One control was discarded as outlier because bias scores were above 6 SD from the mean (group comparison was still significant including this participant). Due to technical error, data from another control was lost. The remaining 71 participants' scores were 95% correct (range = 78–100), with no differences between groups. The groups did not differ in age (001) see .2 = 20.3, d.f. = 20, P = 0.44). Average DSM-IV criteria-count  and CUDIT-scores  decreased in heavy cannabis users (see A 97% follow-up rate was achieved after 6 months (two non-responders among the heavy cannabis users). Within heavy cannabis users, Little's Missing Completely At Random (MCAR ) test wisers see . Cannabisers see .P > 0.12). There was no main effect of image type . The interaction between image type and group was significant . In line with our hypothesis, heavy cannabis users had larger cannabis bias scores than controls , whereas neutral bias scores did not differ between groups . t31 = 2.64, P = 0.013, d = 0.47). This cannabis bias score reflected an approach-bias: approach RTs for cannabis  were faster than for neutral images , whereas avoid RTs for cannabis  and neutral images  did not differ . None of the other bias scores deviated from zero (P > 0.28).Differences in AAT bias scores were first analysed with a mixed ANOVA with group as between-subject factor and image type as within-subject factor with two levels . Homogeneity of variance assumption was not violated  and expectancy factors , but not the compulsivity  and purposefulness factors . The approach-bias correlated positively with weekly cannabis use at 6-month follow-up: higher approach-bias was related to higher levels of cannabis use . The approach-bias did not correlate with post-test craving, baseline cannabis use, baseline and 6-month follow-up measures of cannabis use-related problems and tobacco smoking.In contrast to our hypothesis, the approach-bias correlated negatively with craving in heavy cannabis users: higher approach-bias was related to lower pre-test scores on the MCQ emotionality  was a significant predictor in the first step . The approach-bias explained an additional 13% of the variance in cannabis use 6 months later . Participants with a stronger bias to approach cannabis used more cannabis at 6-month follow-up. Besides the approach-bias, only baseline weekly cannabis use was a significant predictor in the final model. In contrast to our hypothesis, no predictive relations were found between the approach-bias and changes in measures of cannabis-related problems and dependence  after 6 months.To assess the predictive relationship between approach-bias and changes in weekly cannabis use after 6 months (in grams) in heavy users, hierarchical multiple regression was performed. To control for other variables, baseline weekly cannabis use (in grams), CUDIT score, DSM-IV criteria-count, craving factors (pre–post-test average) and neutral bias score were entered first, after which the approach-bias was entered. Preliminary analyses indicated no violation of the assumption of normality, linearity, multicollinearity and homoscedasticity . The total variance explained by the final model was 76% , as measured with the AAT ,29,30. IThe approach-bias found here in heavy cannabis users supports the idea that an approach-bias for substance-related stimuli is a common phenomenon in cannabis users as well as in alcohol users and tobacco smokers ,12,15–17earlier stages of addiction. The approach-bias may predict who will use more, but not who will progress to problematic drug use. This appears to disagree with the incentive sensitization theory of addiction [In contrast to our hypothesis, no associations were found between approach-bias and changes in measures of cannabis dependence. This could be due to methodological issues. Inherent to the sample, cannabis-related problems were relatively low and a 6-month follow-up might have been too short to detect changes in measures of dependence. An alternative explanation is that cognitive biases such as the approach-bias mainly play a role in the course of drug use in the ddiction ,49, and ddiction ,51. To tAlso in contrast to our hypothesis, the approach-bias was associated negatively with pre-test levels of craving for relief from negative affect and craving for anticipation of positive outcome. Post-test craving was not associated with the approach-bias. Compulsive craving predicted cannabis use after 6 months: higher craving was related to increased use. However, the effect disappeared when the approach-bias entered the regression model. Most theories predict a bidirectional positive association between approach-biases and craving . A recenFinally, some limitations must be taken into account. First, there were more tobacco smokers among heavy cannabis users compared to controls, and almost all cannabis users smoked cannabis cigarettes combined with tobacco (most common use-form in the Netherlands ). TobaccIn conclusion, heavy cannabis-smoking young adults automatically activate approach action tendencies in response to cannabis-related stimuli (approach-bias), and the extent to which they do so predicts further escalation of their use.


Salmonella enterica subspecies enterica serovar Enteritidis . Since lipopolysaccharide (LPS) is a major virulence factor and the LPS O-polysaccharide (O-PS) is the immunodominant antigen in serological diagnostic tests, Salmonella mutants lacking O-PS (rough mutants) represent an interesting approach for developing new vaccines and diagnostic tests to differentiate infected and vaccinated animals (DIVA tests). Here, antigenic preparations  from two Salmonella Enteritidis rough mutants, carrying either intact (SEΔwaaL) or deep-defective  LPS-Core, were used in combination with EDA. Biotinylated bacterins, in particular SEΔwaaL bacterin, decorated with EDAvidin (EDA and streptavidin fusion protein) improved the protection conferred by hot-saline or bacterins alone and prevented significantly the virulent infection at least to the levels of live attenuated rough mutants. These findings demonstrate the adjuvant effect of EDAvidin when administered with biotinylated bacterins from Salmonella Enteritidis lacking O-PS and the usefulness of BEDA-SEΔwaaL as non-live vaccine in the mouse model.The Extradomain A from fibronectin (EDA) has an immunomodulatory role as fusion protein with viral and tumor antigens, but its effect when administered with bacteria has not been assessed. Here, we investigated the adjuvant effect of EDA in mice immunizations against  E. coli and tobacco chloroplasts, maintaining its proinflammatory properties  with TLR-3 [E. coli) or MEDA (in transformed plant chloroplast), initially as a simple physical mix with HS extracts (EDA) or bacterins (EDA and MEDA) from Salmonella rough mutants.Once verified that the protection obtained with HS or bacterins from both rough mutants was below that of the B-SEwt, the potential of EDA as immunopotentiator in non-live bacterial vaccines was assessed. Adjuvants such as Freund’s complete or aluminium hydroxide do not appear to improve the immune response against almonella,47, in calmonella,48. Alteth TLR-3 , CpG seqth TLR-3  and bactth TLR-3 ,50. Sincth TLR-3 -8,39, miwaaL or B-SEΔgal). No significant adjuvant effects were observed with EDA or MEDA treatments when B-SEΔwaaL was administered alone , but both EDA or MEDA significantly enhanced protection when using B-SEΔgal. This suggests that the size and/or composition of LPS-Core may have affected the affinity for TLR-4 and/or may have regulated the intracellular fate of the antigen in dendritic cells, as demonstrated for LPS O-PS antigen [gal).The effect of EDA in the efficacy of immunizations depended on the type of antigenic preparation (found in bacterins and not HS extracts) and the mutant from which the bacterins were prepared , the protection conferred by each individual bacterin including SEΔwaaL increased significantly in the presence vs. absence of EDAvidin. Altogether, these findings demonstrate that EDA in the form of EDAvidin-biotin complexes improves the efficacy of non-live vaccines. Like in previous work [waaL and BEDA-SEΔgal), even though the immune response must have been in both cases sufficiently enhanced to confer significant protection.In search of strategies that would help to enhance the binding of EDA to the antigen, the novel recombinant EDA fused to streptavidin molecule (EDAvidin) allowed a significant binding to biotinylated bacterins. Most likely, biotinylated bacterins decorated with EDAvidin enhanced the targeting of LPS defective bacterins to TLR4 expressing cells, modulating the entry of the antigen and/or its intracellular fate and/or the persistence in dendritic cells  to favouous work , increasSalmonella vaccine efficacy use a lethal challenge model. Here, we have used a sub-lethal dose challenge model [waaL performed better than SEΔgal, like in previous lethal challenge reports with similar mutants [gal, and also allowed both the selection of bacterins and not HS from both mutants as Salmonella antigen candidates and the detection of enhanced protection with EDAvidin bound to biotinylated B-SEΔwaaL.Most studies in mice designed to assess ge model  to prese mutants . At the Salmonella bacterins (as biotinylated bacterins) in the mouse model. The association between EDAvidin and B-SEΔwaaL bacterin may be considered safe and effective for use as a non-live vaccine, conferring a high protection against virulent infection. Employing this BEDA immunization strategy with O-PS deficient mutants may also help to distinguish  between vaccinated animals and asymptomatically infected carriers, reservoirs of zoonotic infections. Moreover, the use of non-live vaccines avoids the presence of genetically modified microorganisms in farm animals and their subsequent release to environment or food-chain, having an added value for consumers and veterinary use.Although additional work should be done in different natural hosts to determine the true innocuousness and efficacy of BEDA preparations, it is clear that EDA (as EDAvidin) improves the efficacy of rough The use of EDAvidin is patent pending.Salmonella mutants. BSR, VG, PMM and XDA carried out the microbiological characterization of the mutants, the obtaining and characterization of the Salmonella antigens, the immune response and protection assays in mice, and the statistical analysis. IL and DDA participated in the draft of the manuscript. BA, JJL and MJG conceived the study, participated in its design, coordination, and interpretation of the results, and wrote the manuscript. All authors read and approved the final manuscript.LA, VZ, CM and IF carried out the obtaining and characterization of EDA, MEDA, EDAvidin and biotynilated bacterins. BG carried out the construction and genetic characterization of the waaL in BALB/c mice.Virulence of SEΔgal and SEΔClick here for fileSalmonella Enteritidis parental and mutant strains to cationic peptides and non-immune human serum. (A) Minimal Bactericidal Concentration (MBC) to Polymyxin B; and (B) Susceptibility to conventional human serum, with respect to heat-inactivated serum. Results are expressed as the mean and SD (n = 6) of Polymyxin B concentration (μg/mL) at which bacteria were not recovered (A); and log10 CFU of viable bacteria in fresh serum per million of viable bacteria surviving in heat-inactivated serum (B). Statistical differences (P < 0.01) were found by Fisher’s PLSD test between SE-wt and each rough mutant.Susceptibility of Click here for filewaaL and B-SEwt labeled with carboxyfluorescein succinimidyl ester (CFSE). Unlabeled SE-wt bacterins (B-SEwt) were used as negative control.Flow cytometry of bacterins B-SEΔgal, B-SEΔClick here for fileSalmonella Enteritidis 3934 (SE-wt) strain in mice.Dose–response assay with Click here for file


Reduced kidney function is independently associated with low high-density lipoprotein cholesterol (HDL-C) levels in patients with end-stage renal disease (ESRD), those on hemodialysis, and those with stage 3–5 chronic kidney disease (CKD). However, epidemiological data investigating the relationship between HDL-C levels and kidney function in the general population with roughly normal kidney function are limited, and the results are also inconsistent. The aim of this study was to evaluate the relationship between HDL-C levels and the estimated glomerular filtration rate (eGFR) in a community-based population in China.This was a community-based cross-sectional survey. In total, 4925 participants  were recruited during routine health status examinations. A questionnaire was used to ascertain age, smoking status, and the history of hypertension and diabetes mellitus for each participant. We measured the body mass index, waist circumference, systolic and diastolic blood pressure, and fasting glucose, total cholesterol, triglyceride, HDL-C, low-density lipoprotein cholesterol, uric acid, and serum creatinine level of each participant. eGFR was evaluated using the Chinese modified Modification of Diet in Renal Disease equation.The HDL-C level was higher in the first quartile (lowest quartile) of eGFR than in the fourth quartile (the highest quartile). Additionally, HDL-C levels decreased as eGFR decreased. Pearson’s correlation analysis revealed that HDL-C levels were associated with eGFR (r=0.16). After adjustment for some confounders, HDL-C was independently associated with all quartiles of eGFR in the participants.HDL-C was independently associated with kidney function in a community-dwelling general population. The association between low HDL-C levels and a decreased eGFR gradually strengthened as eGFR declined. Reduced kidney function is an independent risk factor for cardiovascular disease (CVD) and cardiac events . MeanwhiAs an important cardiovascular risk factor, the cardioprotective effects of HDL-C have been confirmed by clinical and experimental research. However, it is unclear whether HDL-C has independent protective effects on kidney function. Epidemiological data investigating the relationship between HDL-C and kidney function in the general population with roughly normal kidney function are limited, and the results are also inconsistent ,9. RecenThe study protocol was approved by the Ethics Committee of the Chinese People’s Liberation Army General Hospital . Each participant provided written informed consent to be included in the study.This community-based cross-sectional survey was designed to establish the relationship between HDL-C and eGFR through routine health status examinations conducted in three communities in Beijing . The study population has been described previously . The comInformation about the age, smoking status, the history of hypertension and diabetes mellitus, and medication use of the participants was obtained using standardized self-report questionnaires. This questionnaire was administered using a face-to-face counseling method. The investigation was completed by physicians in the Department of Geriatric Cardiology of the People’s Liberation Army General Hospital, who were trained by the research team. 2). WHR was calculated as waist circumference (WC) divided by hip circumference. The blood pressure of the participants was measured in the sitting position. The blood pressure measurement was performed using a calibrated desktop sphygmomanometer  after the participants had been in the sitting position for ≥5 min, consistent with current recommendations  independently of the presence of diabetes [2 was 93). The results revealed that low HDL-C levels were independently associated with reduced eGFR in the general population. Some previous studies revealed a relationship between HDL-C and kidney function. However, these studies were overwhelmingly conducted in patients with ESRD or those receiving hemodialysis. Recently, a study reported that 176 patients with CKD , and this independent association was not apparent in the other models . This result implied that other confounders such as age, blood pressure, and lipid parameters may affect kidney function in the general population with roughly normal eGFR values. There are several limitations in the present study. First, because of the cross-sectional design and its inherent limitations, the present study cannot determine causal relationships between the associations. Accordingly, our observations need confirmation in longitudinal and interventional studies. Second, we use creatinine-based eGFR as an index of kidney function. Creatinine is a byproduct of muscle mass, and it is less accurate for assessing kidney function in older adults, who often have reduced muscle mass . There wIn conclusion, HDL-C is independently associated with eGFR after adjustment for multiple covariates in the general population. Moreover, this correlation should be enhanced gradually as eGFR declines. Further, large and well-conducted studies are urgently required to provide more definitive evidence of this correlation.


More insight in the association between reach, dose and fidelity of intervention components and effects is needed. In the current study, we aimed to evaluate reach, dose and fidelity of an individually tailored lifestyle intervention in people with Familial Hypercholesterolemia (FH) and the association between intervention dose and changes in LDL-Cholesterol (LDL-C), and multiple lifestyle behaviours at 12-months follow-up.PRO-FIT*advice) and counselling  using Motivational Interviewing (MI). According to a process evaluation plan, intervention reach, dose delivered and received, and MI fidelity were assessed using the recruitment database, website/counselling logs and the Motivational Interviewing Treatment Integrity (MITI 3.1.1.) code. Regression analyses were conducted to explore differences between participant and non-participant characteristics, and the association between intervention dose and change in LDL-C, and multiple lifestyle behaviours.Participants (n = 181) randomly allocated to the intervention group received the PRO-FIT intervention consisting of computer-tailored lifestyle advice  representative proportion of the intended intervention group was reached during the recruitment phase; participants did not differ from non-participants (n = 623) on age, gender and LDL-C levels. Of the participants, 95% received a PRO-FIT*advice, since it can be relative easily implemented with a high dose delivered. However, only less than half of the intervention group received the complete intervention-package as intended. Strategies to let participants optimally engage in using web-based computer-tailored interventions like PRO-FIT*advice are needed. Further, more emphasis should be put on more extensive MI training and monitoring/supervision.Implementation of the PRO-FIT intervention in practice appears feasible, particularly NTR1899 at ww.trialregister.nl. In public health research, much emphasis is put on the evaluation of interventions in randomised controlled trials (RCTs). Conducting a process evaluation is indispensable, since it helps to explore if the intervention was adopted and implemented as planned, and how and why the intervention worked or not -4. PubliPRO-FIT*advice), and II) counselling  using Motivational Interviewing (MI).In 2009, we started the PRO-FIT project  through an Individually Tailored lifestyle intervention) . The purIn the past years, both computer-tailored lifestyle advice and MI-guided counselling have been tested in RCTs for effects on changes in separate health behaviours. Print-delivered as well as on-line computer-tailored health advice has been shown to be efficacious in changing behaviours, even though effect sizes mostly are small -12. AdvaPRO-FIT*advice, B) face-to-face counselling, C) telephone booster calls, and D) the complete intervention-package, was associated with change in lifestyle behaviour and LDL-C levels    and on L) Figure .st 2007 to April 15th 2009, no longer than 2 years before the start of the project. Access to internet, sufficient fluency in Dutch, residency < 150 km radius from Amsterdam, age 18–70 and LDL-C > 75th percentile were eligibility criteria. People were invited by postal mail and telephoned in case of no response. When people decided to participate, the study procedure was explained by telephone. After randomised allocation to the intervention group, participants were encouraged to visit a weblink referring to the project website, on which they could log on to a personal PRO-FIT*advice account. This account gave access to six tailored advice modules on smoking, physical activity, saturated fat intake, fruit intake, vegetables intake and compliance to statin therapy. Each module required the completion of a screening questionnaire. Subsequently, on-screen personalised feedback was tailored to personal performance level (current lifestyle behaviour), awareness of one’s own performance, as well as personal motivation to change, outcome expectations, attitude and self-efficacy. Personalised feedback to compliance to statin therapy was tailored to knowledge and personal beliefs about (the effect of) statin therapy, potential side effects of the prescribed drug and current compliance. After finishing a module, participants were encouraged to make action plans to change behaviour (except for the advice module on compliance to statin therapy). Thereafter, in a face-to-face session, the participant and the personal coach together further established the level of the participant’s knowledge/awareness about FH and cardiovascular risk factors, according to the assessment(s) and advice(s) within the participant’s personal PRO-FIT*advice account. Ambivalence and barriers related to the recommended behaviour changes were explored in a face-to-face session based on MI techniques. Further, the participant was encouraged to plan five additional counsellor-initiated booster telephone calls, according to their need for additional counselling, intended to support the participant’s behavioural changes and to provide further brief MI to encourage the planned behavioural changes. The two personal coaches had lifestyle coaching and nursing/ teaching backgrounds and had received an additional 3-day MI workshop, incorporating both introductive lessons and practical training sessions with professional actors. A schematic overview of the intervention can be found in Figure Participants of the PRO-FIT trial were recruited from the national cascade screening program of the Foundation for the Identification of Persons with Inherited Hypercholesterolemia (StOEH). Within this program, the StOEH actively approaches first and second degree relatives of index patients  about their potential risk by mail. A genetic field worker telephones and, if the family member agrees to participate, makes an appointment for testing at home. If the results of DNA analysis are positive, first and second degree relatives are approached and offered testing, and so on. No further counseling is given within the screening program. Within tThe RE-AIM evaluation framework conceptualised the evaluation of the translatability of an intervention and included reach, efficacy, adoption, implementation and maintenance . Linnan Guided by Saunders and colleagues, a process evaluation plan was developed in order to monitor and document the implementation of an intervention . In thisIn order to assess the number of people with FH that took part in the project, as well as how representative the participants in the intervention group were for the study population and non-participants , the StOEH client database, as well as the PRO-FIT client database were consulted. Differences between participants and non-participants in main characteristics  were explored.PRO-FIT*advice log on account, a face-to-face counselling session and subsequent telephone booster calls (dose delivered), was assessed by logs that were kept by the coaches and stored in the project database. We aimed at a 100% delivery of the intervention components and delivery of one to five telephone booster calls. The way participants used the PRO-FIT*advice log on account (dose received) was assessed by exploring participants’ log on behaviour (% of participants that logged on), as well as participants’ actions on the PRO-FIT* advice account  by means of log on rates and website use data.The number of participants who had received a http://www.motivationalinterviewing.org), and were trained in coding fidelity using the MITI 3.1.1. For this assessment, a random sample of 20 audio taped counselling sessions  was drawn. A verbatim transcript .The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2458/12/348/prepub


Some patients with depression do not respond to first and second line conventional antidepressants and are therefore characterised as suffering from treatment refractory depression (TRD). On-going psychosocial stress and dysfunction of the hypothalamic-pituitary-adrenal axis are both associated with an attenuated clinical response to antidepressants. Preclinical data shows that co-administration of corticosteroids leads to a reduction in the ability of selective serotonin reuptake inhibitors to increase forebrain 5-hydroxytryptamine, while co-administration of antiglucocorticoids has the opposite effect. A Cochrane review suggests that antiglucocorticoid augmentation of antidepressants may be effective in treating TRD and includes a pilot study of the cortisol synthesis inhibitor, metyrapone. The Antiglucocorticoid augmentation of anti-Depressants in Depression (The ADD Study) is a multicentre randomised placebo controlled trial of metyrapone augmentation of serotonergic antidepressants in a large population of patients with TRD in the UK National Health Service.Patients with moderate to severe treatment refractory Major Depression aged 18 to 65 will be randomised to metyrapone 500 mg twice daily or placebo for three weeks, in addition to on-going conventional serotonergic antidepressants. The primary outcome will be improvement in Montgomery-Åsberg Depression Rating Scale score five weeks after randomisation . Secondary outcomes will include the degree of persistence of treatment effect for up to 6 months, improvements in quality of life and also safety and tolerability of metyrapone. The ADD Study will also include a range of sub-studies investigating the potential mechanism of action of metyrapone.Strengths of the ADD study include broad inclusion criteria meaning that the sample will be representative of patients with TRD treated within the UK National Health Service, longer follow up, which to our knowledge is longer than any previous study of antiglucocorticoid treatments in depression, and the range of mechanistic investigations being carried out. The data set acquired will be a rich resource for a range of research questions relating to both refractory depression and the use of antiglucocorticoid treatments.ISRCTN45338259; EudraCT Number: 2009-015165-31.Current Controlled Trials:  Depression is a common disorder, affecting some 10% of the population . It can 1A autoreceptors and postsynaptic 5-HT receptor efficacy ; NICE: National Institute for Health and Clinical Excellence; NHS: National Health Service; OLM: Object-Location Memory; RCT: Randomised controlled trial; SCID: Structured Clinical Interview for DSM; SSRI: Selective serotonin reuptake inhibitor; STAI: State Trait Anxiety Inventory; SWM: Spatial working memory; TRD: Treatment refractory depression; TSES: Toronto Side Effects Scale; VTL: Verbal Learning Test; YMRS: Young Mania Rating ScaleThe authors declare that they have no competing interests.The study was designed by INF, RHMW, SW, PG, IMA, EMcC NS. Funding was obtained by INF, RHMW, SW, IMA, AOH, EMcC, HCRG, PMH, TH, JW, FHW, AJL, INS. The manuscript was drafted by ES and RHMW. All authors reviewed, revised and approved the final version of the manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-244X/13/205/prepub