Environmental variables drive spatial patterns of trophic diversity in mammals.

Understanding environmental drivers of species diversity has become increasingly important under climate change. Different trophic groups (predators, omnivores and herbivores) interact with their environments in fundamentally different ways and may therefore be influenced by different environmental drivers. Using random forest models, we identified drivers of terrestrial mammals' total and proportional species richness within trophic groups at a global scale. Precipitation seasonality was the most important predictor of richness for all trophic groups. Richness peaked at intermediate precipitation seasonality, indicating that moderate levels of environmental heterogeneity promote mammal richness. Gross primary production (GPP) was the most important correlate of the relative contribution of each trophic group to total species richness. The strong relationship with GPP demonstrates that basal-level resource availability influences how diversity is structured among trophic groups. Our findings suggest that environmental characteristics that influence resource temporal variability and abundance are important predictors of terrestrial mammal richness at a global scale.

Associations of suicide-related media reporting characteristics with help-seeking and suicide in Oregon and Washington.

OBJECTIVE: Specific content characteristics of suicide media reporting might differentially impact suicides in the population, but studies have not considered the overarching theme of the respective media stories and other relevant outcomes besides suicide, such as help-seeking behaviours. METHODS: We obtained 5652 media reports related to suicide from 6 print, 44 broadcast and 251 online sources in Oregon and Washington states, published between April 2019 and March 2020. We conducted a content analysis of stories regarding their overarching focus and specific content characteristics based on media recommendations for suicide reporting. We applied logistic regression analyses to assess how focus and content characteristics were associated with subsequent calls to the US National Suicide Prevention Lifeline (Lifeline) and suicides in these two states in the week after publication compared to a control time period. RESULTS: Compared to a focus on suicide death, a focus on suicidal ideation, suicide prevention, healing stories, community suicide crises/suicide clusters and homicide suicide was associated with more calls. As compared to a focus on suicide death, stories on suicide prevention and stories on community suicide crises/suicide clusters were also associated with no increase in suicides. Regarding specific content characteristics, there were associations that were largely consistent with previous work in the area, for example, an association of celebrity suicide reporting with increases in suicide. CONCLUSION: The overall focus of a media story may influence help-seeking and suicides, and several story characteristics appear to be related to both outcomes. More research is needed to investigate possible causal effects and pathways.

Association of 7 million+ tweets featuring suicide-related content with daily calls to the Suicide Prevention Lifeline and with suicides, United States, 2016-2018.

OBJECTIVE: The aim of this study was to assess associations of various content areas of Twitter posts with help-seeking from the US National Suicide Prevention Lifeline (Lifeline) and with suicides. METHODS: We retrieved 7,150,610 suicide-related tweets geolocated to the United States and posted between 1 January 2016 and 31 December 2018. Using a specially devised machine-learning approach, we categorized posts into content about prevention, suicide awareness, personal suicidal ideation without coping, personal coping and recovery, suicide cases and other. We then applied seasonal autoregressive integrated moving average analyses to assess associations of tweet categories with daily calls to the US National Suicide Prevention Lifeline (Lifeline) and suicides on the same day. We hypothesized that coping-related and prevention-related tweets are associated with greater help-seeking and potentially fewer suicides. RESULTS: The percentage of posts per category was 15.4% (standard deviation: 7.6%) for awareness, 13.8% (standard deviation: 9.4%) for prevention, 12.3% (standard deviation: 9.1%) for suicide cases, 2.4% (standard deviation: 2.1%) for suicidal ideation without coping and 0.8% (standard deviation: 1.7%) for coping posts. Tweets about prevention were positively associated with Lifeline calls (B = 1.94, SE = 0.73, p = 0.008) and negatively associated with suicides (B = -0.11, standard error = 0.05, p = 0.038). Total number of tweets were negatively associated with calls (B = -0.01, standard error = 0.0003, p = 0.007) and positively associated with suicide, (B = 6.4 × 10-5, standard error = 2.6 × 10-5, p = 0.015). CONCLUSION: This is the first large-scale study to suggest that daily volume of specific suicide-prevention-related social media content on Twitter corresponds to higher daily levels of help-seeking behaviour and lower daily number of suicide deaths. PREREGISTRATION: As Predicted, #66922, 26 May 2021.

Spectral binning as an approach to post-acquisition processing of high resolution FIE-MS metabolome fingerprinting data.

INTRODUCTION: Flow infusion electrospray high resolution mass spectrometry (FIE-HRMS) fingerprinting produces complex, high dimensional data sets which require specialist in-silico software tools to process the data prior to analysis. OBJECTIVES: Present spectral binning as a pragmatic approach to post-acquisition procession of FIE-HRMS metabolome fingerprinting data. METHODS: A spectral binning approach was developed that included the elimination of single scan m/z events, the binning of spectra and the averaging of spectra across the infusion profile. The modal accurate m/z was then extracted for each bin. This approach was assessed using four different biological matrices and a mix of 31 known chemical standards analysed by FIE-HRMS using an Exactive Orbitrap. Bin purity and centrality metrics were developed to objectively assess the distribution and position of accurate m/z within an individual bin respectively. RESULTS: The optimal spectral binning width was found to be 0.01 amu. 80.8% of the extracted accurate m/z matched to predicted ionisation products of the chemical standards mix were found to have an error of below 3 ppm. The open-source R package binneR was developed as a user friendly implementation of the approach. This was able to process 100 data files using 4 Central Processing Units (CPU) workers in only 55 seconds with a maximum memory usage of 1.36 GB. CONCLUSION: Spectral binning is a fast and robust method for the post-acquisition processing of FIE-HRMS data. The open-source R package binneR allows users to efficiently process data from FIE-HRMS experiments with the resources available on a standard desktop computer.

FluoroMatch 2.0-making automated and comprehensive non-targeted PFAS annotation a reality.

Because of the pervasiveness, persistence, and toxicity of per- and polyfluoroalkyl substances (PFAS), there is growing concern over PFAS contamination, exposures, and health effects. The diversity of potential PFAS is astounding, with nearly 10,000 PFAS catalogued in databases to date (and growing). The ability to detect the thousands of known PFAS, and discover previously uncatalogued PFAS, is necessary to understand the scope of PFAS contamination and to identify appropriate remediation and regulatory solutions. Current non-targeted methods for PFAS analysis require manual curation and are time-consuming, prone to error, and not comprehensive. FluoroMatch Flow 2.0 is the first software to cover all steps of data processing for PFAS discovery in liquid chromatography-high-resolution tandem mass spectrometry samples. These steps include feature detection, feature blank filtering, exact mass matching to catalogued PFAS, mass defect filtering, homologous series detection, retention time pattern analysis, class-based MS/MS screening, fragment screening, and predicted MS/MS from SMILES structures. In addition, a comprehensive confidence level criterion is implemented to help users understand annotation certainty and integrate various layers of evidence to reduce overreporting. Applying the software to aqueous film forming foam analysis, we discovered over one thousand likely PFAS including previously unreported species. Furthermore, we were able to filter out 96% of features which were likely not PFAS. FluoroMatch Flow 2 increased coverage of likely PFAS by over tenfold compared to the previous release. This software will enable researchers to better characterize PFAS in the environment and in biological systems.

Developing community-based urine sampling methods to deploy biomarker technology for the assessment of dietary exposure.

OBJECTIVE: Obtaining objective, dietary exposure information from individuals is challenging because of the complexity of food consumption patterns and the limitations of self-reporting tools (e.g., FFQ and diet diaries). This hinders research efforts to associate intakes of specific foods or eating patterns with population health outcomes. DESIGN: Dietary exposure can be assessed by the measurement of food-derived chemicals in urine samples. We aimed to develop methodologies for urine collection that minimised impact on the day-to-day activities of participants but also yielded samples that were data-rich in terms of targeted biomarker measurements. SETTING: Urine collection methodologies were developed within home settings. PARTICIPANTS: Different cohorts of free-living volunteers. RESULTS: Home collection of urine samples using vacuum transfer technology was deemed highly acceptable by volunteers. Statistical analysis of both metabolome and selected dietary exposure biomarkers in spot urine collected and stored using this method showed that they were compositionally similar to urine collected using a standard method with immediate sample freezing. Even without chemical preservatives, samples can be stored under different temperature regimes without any significant impact on the overall urine composition or concentration of forty-six exemplar dietary exposure biomarkers. Importantly, the samples could be posted directly to analytical facilities, without the need for refrigerated transport and involvement of clinical professionals. CONCLUSIONS: This urine sampling methodology appears to be suitable for routine use and may provide a scalable, cost-effective means to collect urine samples and to assess diet in epidemiological studies.

Spot and Cumulative Urine Samples Are Suitable Replacements for 24-Hour Urine Collections for Objective Measures of Dietary Exposure in Adults Using Metabolite Biomarkers.

BACKGROUND: Measurement of multiple food intake exposure biomarkers in urine may offer an objective method for monitoring diet. The potential of spot and cumulative urine samples that have reduced burden on participants as replacements for 24-h urine collections has not been evaluated. OBJECTIVE: The aim of this study was to determine the utility of spot and cumulative urine samples for classifying the metabolic profiles of people according to dietary intake when compared with 24-h urine collections in a controlled dietary intervention study. METHODS: Nineteen healthy individuals (10 male, 9 female, aged 21-65 y, BMI 20-35 kg/m2) each consumed 4 distinctly different diets, each for 1 wk. Spot urine samples were collected ∼2 h post meals on 3 intervention days/wk. Cumulative urine samples were collected daily over 3 separate temporal periods. A 24-h urine collection was created by combining the 3 cumulative urine samples. Urine samples were analyzed with metabolite fingerprinting by both high-resolution flow infusion electrospray mass spectrometry (FIE-HRMS) and proton nuclear magnetic resonance spectroscopy (1H-NMR). Concentrations of dietary intake biomarkers were measured with liquid chromatography triple quadrupole mass spectrometry and by integration of 1H-NMR data. RESULTS: Cross-validation modeling with 1H-NMR and FIE-HRMS data demonstrated the power of spot and cumulative urine samples in predicting dietary patterns in 24-h urine collections. Particularly, there was no significant loss of information when post-dinner (PD) spot or overnight cumulative samples were substituted for 24-h urine collections (classification accuracies of 0.891 and 0.938, respectively). Quantitative analysis of urine samples also demonstrated the relation between PD spot samples and 24-h urines for dietary exposure biomarkers. CONCLUSIONS: We conclude that PD spot urine samples are suitable replacements for 24-h urine collections. Alternatively, cumulative samples collected overnight predict similarly to 24-h urine samples and have a lower collection burden for participants.

Integrated Analytical and Statistical Two-Dimensional Spectroscopy Strategy for Metabolite Identification: Application to Dietary Biomarkers.

A major purpose of exploratory metabolic profiling is for the identification of molecular species that are statistically associated with specific biological or medical outcomes; unfortunately, the structure elucidation process of unknowns is often a major bottleneck in this process. We present here new holistic strategies that combine different statistical spectroscopic and analytical techniques to improve and simplify the process of metabolite identification. We exemplify these strategies using study data collected as part of a dietary intervention to improve health and which elicits a relatively subtle suite of changes from complex molecular profiles. We identify three new dietary biomarkers related to the consumption of peas (N-methyl nicotinic acid), apples (rhamnitol), and onions (N-acetyl-S-(1Z)-propenyl-cysteine-sulfoxide) that can be used to enhance dietary assessment and assess adherence to diet. As part of the strategy, we introduce a new probabilistic statistical spectroscopy tool, RED-STORM (Resolution EnhanceD SubseT Optimization by Reference Matching), that uses 2D J-resolved 1H NMR spectra for enhanced information recovery using the Bayesian paradigm to extract a subset of spectra with similar spectral signatures to a reference. RED-STORM provided new information for subsequent experiments (e.g., 2D-NMR spectroscopy, solid-phase extraction, liquid chromatography prefaced mass spectrometry) used to ultimately identify an unknown compound. In summary, we illustrate the benefit of acquiring J-resolved experiments alongside conventional 1D 1H NMR as part of routine metabolic profiling in large data sets and show that application of complementary statistical and analytical techniques for the identification of unknown metabolites can be used to save valuable time and resources.

Diversity and association of phenotypic and metabolomic traits in the close model grasses Brachypodium distachyon, B. stacei and B. hybridum.

Background and Aims: Morphological traits in combination with metabolite fingerprinting were used to investigate inter- and intraspecies diversity within the model annual grasses Brachypodium distachyon, Brachypodium stacei and Brachypodium hybridum . Methods: Phenotypic variation of 15 morphological characters and 2219 nominal mass ( m / z ) signals generated using flow infusion electrospray ionization-mass spectrometry (FIE-MS) were evaluated in individuals from a total of 174 wild populations and six inbred lines, and 12 lines, of the three species, respectively. Basic statistics and multivariate principal component analysis and discriminant analysis were used to differentiate inter- and intraspecific variability of the two types of variable, and their association was assayed with the rcorr function. Key Results: Basic statistics and analysis of variance detected eight phenotypic characters [(stomata) leaf guard cell length, pollen grain length, (plant) height, second leaf width, inflorescence length, number of spikelets per inflorescence, lemma length, awn length] and 434 tentatively annotated metabolite signals that significantly discriminated the three species. Three phenotypic traits (pollen grain length, spikelet length, number of flowers per inflorescence) might be genetically fixed. The three species showed different metabolomic profiles. Discriminant analysis significantly discriminated the three taxa with both morphometric and metabolome traits and the intraspecific phenotypic diversity within B. distachyon and B. stacei . The populations of B. hybridum were considerably less differentiated. Conclusions: Highly explanatory metabolite signals together with morphological characters revealed concordant patterns of differentiation of the three taxa. Intraspecific phenotypic diversity was observed between northern and southern Iberian populations of B. distachyon and between eastern Mediterranean/south-western Asian and western Mediterranean populations of B. stacei . Significant association was found for pollen grain length and lemma length and ten and six metabolomic signals, respectively. These results would guide the selection of new germplasm lines of the three model grasses in ongoing genome-wide association studies.

Efficacy of the TuttleNumbNow Intraosseous Method for Pulpal Anesthesia in the Mandibular First Molar: A Prospective, Randomized, Crossover Study.

INTRODUCTION: Previous studies on intraosseous (IO) anesthesia as a primary injection have shown high success rates. The TuttleNumbNow (TNN; Orem, UT) is a new primary IO injection technique that has not been scientifically evaluated. Therefore, the purpose of this prospective randomized, crossover study was to evaluate the anesthetic efficacy of the TNN IO technique using the Septoject Evolution needle (Septodont, Saint-Maur-des-Fosses, France) compared with buccal infiltration for pulpal anesthesia in mandibular first molars. METHODS: One hundred four healthy subjects were randomly assigned to 2 treatment groups separated by at least 2 weeks. One set of injections consisted of buccal infiltration of the mandibular first molar using 1.8 mL 4% articaine with 1:100,000 epinephrine followed by a mock TNN injection distal to the mandibular first molar. The other set of injections was a mock buccal infiltration of the mandibular first molar followed by a TNN injection of 1.8 mL 4% articaine with 1:100,000 epinephrine distal to the mandibular first molar. Statistical analyses were performed. RESULTS: For the mandibular first molar, which had a 42% anesthetic success rate (highest 80 reading) with buccal infiltration compared with 49% with the TNN, no statistically significant difference in success was observed (P = .2115). CONCLUSIONS: The TNN technique has been advocated as an IO injection. However, the inability to deliver anesthetic solution to the cancellous bone resulted in an anesthetic success rate of 49%. The success was statistically similar to a buccal infiltration (42%) and would not provide adequate pulpal anesthesia as a primary injection.

"Breaking the Silence" Suicide Prevention Media Campaign in Oregon, April 7-14, 2019.

Background: Between April 7 and 14, 2019, the "Breaking the Silence" media engagement campaign was launched in Oregon. Aims: We aimed to assess the consistency of media content related to the campaign with media guidelines and the quantitative footprint on Twitter (now X) over time. Method: Media items related to the campaign were analyzed regarding focus and consistency with media guidelines for suicide reporting and compared with other suicide-related reports published in the same time frame, as well as with reporting in Washington, the control region. Tweets related to the campaign were retrieved to assess the social media footprint. Results: There were n = 104 media items in the campaign month, mainly in the campaign week. Items typically used a narrative featuring suicide advocacy or policy/prevention programs. As compared to other items with a similar focus, they scored better on several protective characteristics listed in media recommendations. Stories of coping with adversity, however, were scarce. The social media footprint on Twitter was small. Limitations: Inability to make causal claims about campaign impact. Conclusion: Media items from the Breaking the Silence campaign appeared mainly consistent with media guidelines, but some aspects, such as stories of recovery, were under-represented.

Metabolic changes in response to varying whole-grain wheat and rye intake.

Epidemiological studies have shown associations between whole-grain intake and lowered disease risk. A sufficient level of whole-grain intake to reach the health benefits has not been established, and there is limited knowledge about the impact of whole-grain intake on metabolite levels. In this clinical intervention study, we aimed to identify plasma and urine metabolites associated with two different intake levels of whole-grain wheat and rye and to correlate them with clinical plasma biomarkers. Healthy volunteers (N = 68) were divided into two groups receiving either whole-grain wheat or whole-grain rye in two four-week interventions with 48 and 96 g/d of whole grains consumed. The metabolomics of the plasma samples was performed with UPLC-QTOF-MS. Plasma alkylresorcinols were quantified with GC-MS and plasma and urinary mammalian lignans with HPLC-ECD. The high-dose intervention impacted the metabolite profile, including microbial metabolites, more in the rye-enriched diet compared with wheat. Among the increased metabolites were alkylresorcinol glucuronides, sinapyl alcohol, and pipecolic acid betaine, while the decreased metabolites included acylcarnitines and ether lipids. Plasma alkylresorcinols, urinary enterolactone, and total mammalian lignans reflected the study diets in a dose-dependent manner. Several key metabolites linked with whole-grain consumption and gut microbial metabolism increased in a linear manner between the two interventions. The results reveal that an increase in whole-grain intake, particularly rye, is strongly reflected in the metabolite profile, is correlated with clinical variables, and suggests that a diet rich in whole grains promotes the growth and/or metabolism of microbes producing potentially beneficial microbial metabolites.

Deciphering systemic wound responses of the pumpkin extrafascicular phloem by metabolomics and stable isotope-coded protein labeling.

In cucurbits, phloem latex exudes from cut sieve tubes of the extrafascicular phloem (EFP), serving in defense against herbivores. We analyzed inducible defense mechanisms in the EFP of pumpkin (Cucurbita maxima) after leaf damage. As an early systemic response, wounding elicited transient accumulation of jasmonates and a decrease in exudation probably due to partial sieve tube occlusion by callose. The energy status of the EFP was enhanced as indicated by increased levels of ATP, phosphate, and intermediates of the citric acid cycle. Gas chromatography coupled to mass spectrometry also revealed that sucrose transport, gluconeogenesis/glycolysis, and amino acid metabolism were up-regulated after wounding. Combining ProteoMiner technology for the enrichment of low-abundance proteins with stable isotope-coded protein labeling, we identified 51 wound-regulated phloem proteins. Two Sucrose-Nonfermenting1-related protein kinases and a 32-kD 14-3-3 protein are candidate central regulators of stress metabolism in the EFP. Other proteins, such as the Silverleaf Whitefly-Induced Protein1, Mitogen Activated Protein Kinase6, and Heat Shock Protein81, have known defensive functions. Isotope-coded protein labeling and western-blot analyses indicated that Cyclophilin18 is a reliable marker for stress responses of the EFP. As a hint toward the induction of redox signaling, we have observed delayed oxidation-triggered polymerization of the major Phloem Protein1 (PP1) and PP2, which correlated with a decline in carbonylation of PP2. In sum, wounding triggered transient sieve tube occlusion, enhanced energy metabolism, and accumulation of defense-related proteins in the pumpkin EFP. The systemic wound response was mediated by jasmonate and redox signaling.

Regulatory barriers to dental anesthesia services for special populations.

AIMS: Sedation and general anesthesia are necessities for the treatment of many individuals within special populations such as those with physical and intellectual disabilities, fear/anxiety, or individuals requiring extensive procedures. This study aims to discover regulatory factors that may be contributing to the limited access to anesthesia services provided by dentist anesthesiologists. METHODS AND RESULTS: The study included an online survey completed by self-reported dentist anesthesiologists with 2 or more years of formal anesthesia training. The survey was distributed at the April 2019 American Society of Dentist Anesthesiologists national meeting in Chicago. Participants responded to questions regarding the effect of specific state regulations on decisions to practice in a particular state and how such regulations influenced patient safety and barriers to care. Rules and regulatory restrictions on the mobility of dentist anesthesiologist equipment/supplies and additional state narcotic transportation regulations were deemed statistically significant in failing to improve safety. Requiring airway and sedation training for a facility's provider and staff were not barriers to care. Rules and regulations were not a factor to establishing clinical practice in one state over another state. CONCLUSION: Individuals and organizations responsible for influencing the regulatory environment of anesthesia services should improve regulations to facilitate the mobility of dentist anesthesiologists.

Optimising peace through a Universal Global Peace Treaty to constrain the risk of war from a militarised artificial superintelligence.

This article argues that an artificial superintelligence (ASI) emerging in a world where war is still normalised constitutes a catastrophic existential risk, either because the ASI might be employed by a nation-state to war for global domination, i.e., ASI-enabled warfare, or because the ASI wars on behalf of itself to establish global domination, i.e., ASI-directed warfare. Presently, few states declare war or even war on each other, in part due to the 1945 UN Charter, which states Member States should "refrain in their international relations from the threat or use of force", while allowing for UN Security Council-endorsed military measures and self-defense. As UN Member States no longer declare war on each other, instead, only 'international armed conflicts' occur. However, costly interstate conflicts, both hot and cold and tantamount to wars, still take place. Further, a New Cold War between AI superpowers looms. An ASI-directed/enabled future conflict could trigger total war, including nuclear conflict, and is therefore high risk. Via conforming instrumentalism, an international relations theory, we advocate risk reduction by optimising peace through a Universal Global Peace Treaty (UGPT), contributing towards the ending of existing wars and prevention of future wars, as well as a Cyberweapons and Artificial Intelligence Convention. This strategy could influence state actors, including those developing ASIs, or an agential ASI, particularly if it values conforming instrumentalism and peace. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00146-021-01382-y.

Beating the heat: ecology of desert bobcats.

BACKGROUND: Relative to temperate regions, little is known about bobcats (Lynx rufus) in the Sonoran Desert portion of their range, in part due to the difficulty of sampling an elusive carnivore in harsh desert environments. Here, we quantify habitat selection and evaluate diet of bobcats at Kofa National Wildlife Refuge, Arizona, USA, using multiple sampling techniques including GPS telemetry, camera traps, and DNA metabarcoding. RESULTS: Home ranges during the hot season were smaller than during the cool season. Camera trapping failed to yield a high enough detection rate to identify habitat occupancy trends but third-order resource selection from GPS-collar data showed a preference for higher elevations and rugged terrain at lower elevations. Diet composition consisted of a diverse range of available small prey items, including a higher frequency of avian prey than previously observed in bobcats. CONCLUSIONS: Desert bobcats in our study maintained smaller home ranges and primarily consumed smaller prey than their more northern relatives. This study illustrates the benefit of employing multiple, complementary sampling methods to understand the ecology of elusive species.

Prospective Study on PDL Anesthesia as an Aide to Decrease Palatal Infiltration Pain.

OBJECTIVE: The purpose of this prospective randomized study was to assess using a periodontal ligament (PDL) injection as an aide to decrease palatal infiltration pain. METHODS: A total of 133 subjects randomly received a PDL injection and alternative palatal infiltration or a mock PDL injection and conventional palatal infiltration at 2 separate appointments. PDL injection was given in the mid-palatal sulcus of the maxillary first molar. Mock PDL injection consisted of only needle insertion. All subjects then received a palatal infiltration administered into the blanched gingival tissue 3 mm (alternative palatal infiltration) or 7 mm (conventional palatal infiltration) from the gingival collar. Subjects recorded needle insertion and solution deposition pain using a Heft-Parker visual analog scale (VAS). RESULTS: The combined PDL injection and alternative palatal infiltration had significantly decreased mean VAS ratings for needle insertion and solution deposition pain (P < .0001). Incidence of moderate/severe pain for needle insertion and solution deposition was reduced from 65% to 1% and from 65% to 2%, respectively. CONCLUSION: Providing PDL anesthesia into the mid-palatal sulcus of the maxillary first molar and then administering an alternative palatal infiltration into the blanched collar around the PDL molar site led to significant reductions in needle insertion and solution deposition pain compared with a mock PDL and conventional palatal infiltration.

Assessing Adherence to Healthy Dietary Habits Through the Urinary Food Metabolome: Results From a European Two-Center Study.

Background: Diet is one of the most important modifiable lifestyle factors in human health and in chronic disease prevention. Thus, accurate dietary assessment is essential for reliably evaluating adherence to healthy habits. Objectives: The aim of this study was to identify urinary metabolites that could serve as robust biomarkers of diet quality, as assessed through the Alternative Healthy Eating Index (AHEI-2010). Design: We set up two-center samples of 160 healthy volunteers, aged between 25 and 50, living as a couple or family, with repeated urine sampling and dietary assessment at baseline, and 6 and 12 months over a year. Urine samples were subjected to large-scale metabolomics analysis for comprehensive quantitative characterization of the food-related metabolome. Then, lasso regularized regression analysis and limma univariate analysis were applied to identify those metabolites associated with the AHEI-2010, and to investigate the reproducibility of these associations over time. Results: Several polyphenol microbial metabolites were found to be positively associated with the AHEI-2010 score; urinary enterolactone glucuronide showed a reproducible association at the three study time points [false discovery rate (FDR): 0.016, 0.014, 0.016]. Furthermore, other associations were found between the AHEI-2010 and various metabolites related to the intake of coffee, red meat and fish, whereas other polyphenol phase II metabolites were associated with higher AHEI-2010 scores at one of the three time points investigated (FDR < 0.05 or ß ≠ 0). Conclusion: We have demonstrated that urinary metabolites, and particularly microbiota-derived metabolites, could serve as reliable indicators of adherence to healthy dietary habits. Clinical Trail Registration: www.ClinicalTrials.gov, Identifier: NCT03169088.

The metabolic transition during disease following infection of Arabidopsis thaliana by Pseudomonas syringae pv. tomato.

The outcome of bacterial infection in plants is determined by the ability of the pathogen to successfully occupy the apoplastic space and deliver a constellation of effectors that collectively suppress basal and effector-triggered immune responses. In this study, we examined the metabolic changes associated with establishment of disease using analytical techniques that interrogated a range of chemistries. We demonstrated clear differences in the metabolome of Arabidopsis thaliana leaves infected with virulent Pseudomonas syringae within 8 h of infection. In addition to confirmation of changes in phenolic and indolic compounds, we identified rapid alterations in the abundance of amino acids and other nitrogenous compounds, specific classes of glucosinolates, disaccharides, and molecules that influence the prevalence of reactive oxygen species. Our data illustrate that, superimposed on defence suppression, pathogens reconfigure host metabolism to provide the sustenance required to support exponentially growing populations of apoplastically localized bacteria. We performed a detailed baseline study reporting the metabolic dynamics associated with bacterial infection. Moreover, we have integrated these data with the results of transcriptome profiling to distinguish metabolomic pathways that are transcriptionally activated from those that are post-transcriptionally regulated.