Integrative Molecular Phenotyping
INTEGRATIVE MOLECULAR
PHENOTYPING
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY

PubMed

Personalised Interventions-A Precision Approach for the Next Generation of Dietary Intervention Studies.

Thu, 10/08/2017 - 13:58
Related Articles Personalised Interventions-A Precision Approach for the Next Generation of Dietary Intervention Studies. Nutrients. 2017 Aug 09;9(8): Authors: de Roos B, Brennan L Abstract Diet is a key modifiable risk factor for non-communicable diseases. However, we currently are not benefitting from the full potential of its protective effects. This is due to a number of reasons, including high individual variability in response to certain diets. It is now well acknowledged that in order to gain the full benefit of dietary regimes it is essential to take into account individual responses. With this in mind, the present review examines the concept of precision nutrition and the performance of n-of-1 studies, and discusses the development of certain approaches that will be critical for development of the concepts. PMID: 28792454 [PubMed - in process]

Special Issue: Cancer Metabolism.

Thu, 10/08/2017 - 13:58
Related Articles Special Issue: Cancer Metabolism. Metabolites. 2017 Aug 09;7(3): Authors: Basetti M Abstract This special issue is designed to present the latest research findings and developments in the field of cancer metabolism. Cancer is a complex disease and a common term used for more than 100 diseases, whereas metabolism describes a labyrinth of complex biochemical pathways in the cell. It is essential to understand metabolism in the context of cancer for the early detection of disease biomarkers and to find proper targets for potential treatments. The articles presented in this issue cover metabolic aspects of brain tumours, breast tumours, paraganglioma, and the metabolic activity of tumour suppressor gene p53. PMID: 28792436 [PubMed]

Advances in the analysis of "less-conventional" human body fluids: an overview of the ce- and hplc-ms applications in the years 2015-2017.

Thu, 10/08/2017 - 13:58
Related Articles Advances in the analysis of "less-conventional" human body fluids: an overview of the ce- and hplc-ms applications in the years 2015-2017. Electrophoresis. 2017 Aug 09;: Authors: Venere MD, Viglio S, Cagnone M, Bardoni A, Salvini R, Iadarola P Abstract Aim of this article is to focus the attention of the reader on the application of CE/MS and LC/MS to the analysis of human body fluids not currently used for the diagnosis of disorders and, for this reason, catalogued as "less-/non-conventional" fluids, i.e. tears, nasal secretions, cerumen, bronchoalveolar lavage fluid, sputum, exhaled breath condensate, nipple aspirate, breast milk, amniotic fluid, bile, seminal plasma, liposuction aspirate fluid and synovial fluid. The pool of articles presented in this report demonstrates that, rather than being neglected, these fluids are an important resource for the evaluation of possible pathologic conditions. Thus, being a sort of mirror that reflects the normal internal characteristics and disease state of an individual, they benefit of an increasing appreciation. This review follows a previous report of this series and covers the latest developments in this field which have been published in specialist journals in the years 2015-2017. This article is protected by copyright. All rights reserved. PMID: 28792066 [PubMed - as supplied by publisher]

Ambient mass spectrometry in metabolomics.

Thu, 10/08/2017 - 13:58
Related Articles Ambient mass spectrometry in metabolomics. Analyst. 2017 Aug 09;: Authors: Clendinen CS, Monge ME, Fernández FM Abstract Since the introduction of desorption electrospray ionization (DESI) mass spectrometry (MS), ambient MS methods have seen increased use in a variety of fields from health to food science. Increasing its popularity in metabolomics, ambient MS offers limited sample preparation, rapid and direct analysis of liquids, solids, and gases, in situ and in vivo analysis, and imaging. The metabolome consists of a constantly changing collection of small (<1.5 kDa) molecules. These include endogenous molecules that are part of primary metabolism pathways, secondary metabolites with specific functions such as signaling, chemicals incorporated in the diet or resulting from environmental exposures, and metabolites associated with the microbiome. Characterization of the responsive changes of this molecule cohort is the principal goal of any metabolomics study. With adjustments to experimental parameters, metabolites with a range of chemical and physical properties can be selectively desorbed and ionized and subsequently analyzed with increased speed and sensitivity. This review covers the broad applications of a variety of ambient MS techniques in four primary fields in which metabolomics is commonly employed. PMID: 28792022 [PubMed - as supplied by publisher]

Hyphenated MS-based targeted approaches in metabolomics.

Thu, 10/08/2017 - 13:58
Related Articles Hyphenated MS-based targeted approaches in metabolomics. Analyst. 2017 Aug 09;: Authors: Begou O, Gika HG, Wilson ID, Theodoridis G Abstract While global metabolic profiling (untargeted metabolomics) has been the center of much interest and research activity in the past few decades, more recently targeted metabolomics approaches have begun to gain ground. These analyses are, to an extent, more hypothesis-driven, as they focus on a set of pre-defined metabolites and aim towards their determination, often to the point of absolute quantification. The continuous development of the technological platforms used in these studies facilitates the analysis of large numbers of well-characterized metabolites present in complex matrices. The present review describes recent developments in the hyphenated chromatographic methods most often applied in targeted metabolomic/lipidomic studies (LC-MS/MS, CE-MS/MS, and GC-MS/MS), highlighting applications in the life and food/plant sciences. The review also underlines practical challenges-limitations that appear in such approaches. PMID: 28792021 [PubMed - as supplied by publisher]

Evolution of newborns urinary metabolomic profiles according to age and growth.

Thu, 10/08/2017 - 13:58
Related Articles Evolution of newborns urinary metabolomic profiles according to age and growth. J Proteome Res. 2017 Aug 09;: Authors: Scalabre A, Jobard E, Demède D, Gaillard S, Pontoizeau C, Mouriquand P, Elena-Herrmann B, Mure PY Abstract Improving the management of neonatal diseases and prevention of chronic diseases in adulthood requires a better comprehension of the complex maturational processes associated with newborns development. Urine-based metabolomic studies play a promising role in the fields of pediatrics and neonatology, relying on simple and non-invasive collection procedures while integrating a variety of factors such as genotype, nutritional state, lifestyle and diseases. Here, we investigate the influence of age, weight, height and gender on the urine metabolome during the first 4 months of life. Untargeted analysis of urine was carried out by 1H-Nuclear Magnetic Resonance (NMR) spectroscopy for 90 newborns under 4 months of age, and free of metabolic, nephrologic or urologic diseases. Supervised multivariate statistical analysis of the metabolic profiles revealed metabolites significantly associated with age, weight and height respectively. The tremendous growth occurring during the neonatal period is associated with specific modifications of newborns metabolism. Conversely, gender appears to have no impact on the urine metabolome during early infancy. These results allow a deeper understanding of newborn metabolic maturation and underline potential confounding factors in newborns metabolomics studies. We emphasize the need to systematically and precisely report children age, height and weight that impact urine metabolic profiles of infants. PMID: 28791867 [PubMed - as supplied by publisher]

Metabolomics and Proteomics of Brassica napus Guard Cells in Response to Low CO2.

Thu, 10/08/2017 - 13:58
Related Articles Metabolomics and Proteomics of Brassica napus Guard Cells in Response to Low CO2. Front Mol Biosci. 2017;4:51 Authors: Geng S, Yu B, Zhu N, Dufresne C, Chen S Abstract Stomatal guard cell response to various stimuli is an important process that balances plant carbon dioxide (CO2) uptake and water transpiration. Elevated CO2 induces stomatal closure, while low CO2 promotes stomatal opening. The signaling process of elevated CO2 induced stomatal closure has been extensively studied in recent years. However, the mechanism of low CO2 induced stomatal opening is not fully understood. Here we report metabolomic and proteomic responses of Brassica napus guard cells to low CO2 using hyphenated mass spectrometry technologies. A total of 411 metabolites and 1397 proteins were quantified in a time-course study of low CO2 effects. Metabolites and proteins that exhibited significant changes are overrepresented in fatty acid metabolism, starch and sucrose metabolism, glycolysis and redox regulation. Concomitantly, multiple hormones that promote stomatal opening increased in response to low CO2. Interestingly, jasmonic acid precursors were diverted to a branch pathway of traumatic acid biosynthesis. These results indicate that the low CO2 response is mediated by a complex crosstalk between different phytohormones. PMID: 28791296 [PubMed]

Gut Microbiome Response to Sucralose and Its Potential Role in Inducing Liver Inflammation in Mice.

Thu, 10/08/2017 - 13:58
Related Articles Gut Microbiome Response to Sucralose and Its Potential Role in Inducing Liver Inflammation in Mice. Front Physiol. 2017;8:487 Authors: Bian X, Chi L, Gao B, Tu P, Ru H, Lu K Abstract Sucralose is the most widely used artificial sweetener, and its health effects have been highly debated over the years. In particular, previous studies have shown that sucralose consumption can alter the gut microbiota. The gut microbiome plays a key role in processes related to host health, such as food digestion and fermentation, immune cell development, and enteric nervous system regulation. Inflammation is one of the most common effects associated with gut microbiome dysbiosis, which has been linked to a series of human diseases, such as diabetes and obesity. The aim of this study was to investigate the structural and functional effects of sucralose on the gut microbiota and associated inflammation in the host. In this study, C57BL/6 male mice received sucralose in their drinking water for 6 months. The difference in gut microbiota composition and metabolites between control and sucralose-treated mice was determined using 16S rRNA gene sequencing, functional gene enrichment analysis and metabolomics. Inflammatory gene expression in tissues was analyzed by RT-PCR. Alterations in bacterial genera showed that sucralose affects the gut microbiota and its developmental dynamics. Enrichment of bacterial pro-inflammatory genes and disruption in fecal metabolites suggest that 6-month sucralose consumption at the human acceptable daily intake (ADI) may increase the risk of developing tissue inflammation by disrupting the gut microbiota, which is supported by elevated pro-inflammatory gene expression in the liver of sucralose-treated mice. Our results highlight the role of sucralose-gut microbiome interaction in regulating host health-related processes, particularly chronic inflammation. PMID: 28790923 [PubMed]

Personomics and Precision Medicine.

Thu, 10/08/2017 - 13:58
Related Articles Personomics and Precision Medicine. Trans Am Clin Climatol Assoc. 2017;128:160-168 Authors: Ziegelstein RC Abstract The importance of knowing patients as individuals has been highlighted throughout the history of medicine. However, shorter visits, electronic documentation, reliance on technology, and increasing linguistic and cultural differences between patients and physicians create more challenges to effective communication than ever before. Perhaps more concerning is the greater emphasis on aspects of care considered more precisely measurable and quantifiable, the sum of which is sometimes felt to represent the patient better than knowledge of the patient himself. While genomics, proteomics, pharmacogenomics, metabolomics, and epigenomics promise enhanced diagnostics and therapeutics, understanding the unique circumstances of the person - what may be called personomics - is at least as critical to patient care. Such an understanding can only be developed when the relevant psychological, social, cultural, behavioral, and economic factors are obtained. Personomics determines how a disease reveals itself phenotypically and the way that disease and the individual with the disease respond to treatment. PMID: 28790500 [PubMed - in process]

Cellobiose Consumption Uncouples Extracellular Glucose Sensing and Glucose Metabolism in Saccharomyces cerevisiae.

Thu, 10/08/2017 - 13:58
Related Articles Cellobiose Consumption Uncouples Extracellular Glucose Sensing and Glucose Metabolism in Saccharomyces cerevisiae. MBio. 2017 Aug 08;8(4): Authors: Chomvong K, Benjamin DI, Nomura DK, Cate JHD Abstract Glycolysis is central to energy metabolism in most organisms and is highly regulated to enable optimal growth. In the yeast Saccharomyces cerevisiae, feedback mechanisms that control flux through glycolysis span transcriptional control to metabolite levels in the cell. Using a cellobiose consumption pathway, we decoupled glucose sensing from carbon utilization, revealing new modular layers of control that induce ATP consumption to drive rapid carbon fermentation. Alterations of the beta subunit of phosphofructokinase-1 (PFK2), H(+)-plasma membrane ATPase (PMA1), and glucose sensors (SNF3 and RGT2) revealed the importance of coupling extracellular glucose sensing to manage ATP levels in the cell. Controlling the upper bound of cellular ATP levels may be a general mechanism used to regulate energy levels in cells, via a regulatory network that can be uncoupled from ATP concentrations under perceived starvation conditions.IMPORTANCE Living cells are fine-tuned through evolution to thrive in their native environments. Genome alterations to create organisms for specific biotechnological applications may result in unexpected and undesired phenotypes. We used a minimal synthetic biological system in the yeast Saccharomyces cerevisiae as a platform to reveal novel connections between carbon sensing, starvation conditions, and energy homeostasis. PMID: 28790206 [PubMed - in process]

Metabolic and RNA profiling elucidates proanthocyanidins accumulation in Aglianico grape.

Thu, 10/08/2017 - 13:58
Related Articles Metabolic and RNA profiling elucidates proanthocyanidins accumulation in Aglianico grape. Food Chem. 2017 Oct 15;233:52-59 Authors: Rinaldi A, Villano C, Lanzillo C, Tamburrino A, Jourdes M, Teissedre PL, Moio L, Frusciante L, Carputo D, Aversano R Abstract Aglianico grapes are known for their high content of proanthocyanidins (PAs), which are responsible for the astringency of wines derived from this cultivar. However, the accumulation of PAs and their genetic control during berry development remain largely unexplored. This work aimed to monitor astringency-causing PAs in Aglianico berries and correlate them with the expression of 14 key genes. Berries were collected during ripening and dissected in skins and seeds. PAs were fractionated and the content of total phenolics, flavans, anthocyanins, tannins reactive towards salivary proteins and tannin structural composition were evaluated. The results provided evidence that PAs were more abundant in seeds than in skins as expected, with differences in the structural composition between tissues, which did not varied during ripening. Expression analysis showed that Aglianico is able to accumulate polyphenols due to its ability to modulate key genes in a tissue-specific manner. PMID: 28530607 [PubMed - indexed for MEDLINE]

The anti-cancerous drug doxorubicin decreases the c-di-GMP content in Pseudomonas aeruginosa but promotes biofilm formation.

Thu, 10/08/2017 - 13:58
Related Articles The anti-cancerous drug doxorubicin decreases the c-di-GMP content in Pseudomonas aeruginosa but promotes biofilm formation. Microbiology. 2016 Oct;162(10):1797-1807 Authors: Groizeleau J, Rybtke M, Andersen JB, Berthelsen J, Liu Y, Yang L, Nielsen TE, Kaever V, Givskov M, Tolker-Nielsen T Abstract Current antibiotic treatments are insufficient in eradicating bacterial biofilms, which represent the primary cause of chronic bacterial infections. Thus, there is an urgent need for new strategies to eradicate biofilm infections. The second messenger c-di-GMP is a positive regulator of biofilm formation in many clinically relevant bacteria. It is hypothesized that drugs lowering the intracellular level of c-di-GMP will force biofilm bacteria into a more treatable planktonic lifestyle. To identify compounds capable of lowering c-di-GMP levels in Pseudomonas aeruginosa, we screened 5000 compounds for their potential c-di-GMP-lowering effect using a recently developed c-di-GMP biosensor strain. Our screen identified the anti-cancerous drug doxorubicin as a potent c-di-GMP inhibitor. In addition, the drug decreased the transcription of many biofilm-related genes. However, despite its effect on the c-di-GMP content in P. aeruginosa, doxorubicin was unable to inhibit biofilm formation or disperse established biofilms. On the contrary, the drug was found to promote P. aeruginosa biofilm formation, possibly through release of extracellular DNA from a subpopulation of killed bacteria. Our findings emphasize that lowering of the c-di-GMP content in bacteria might not be sufficient to mediate biofilm inhibition or dispersal. PMID: 27526691 [PubMed - indexed for MEDLINE]

Energy and lipid metabolism during direct and diapause development in a pierid butterfly.

Thu, 10/08/2017 - 13:58
Related Articles Energy and lipid metabolism during direct and diapause development in a pierid butterfly. J Exp Biol. 2016 Oct 01;219(Pt 19):3049-3060 Authors: Lehmann P, Pruisscher P, Posledovich D, Carlsson M, Käkelä R, Tang P, Nylin S, Wheat CW, Wiklund C, Gotthard K Abstract Diapause is a fundamental component of the life cycle in the majority of insects living in environments characterized by strong seasonality. The present study addresses poorly understood associations and trade-offs between endogenous diapause duration, thermal sensitivity of development, energetic cost of development and cold tolerance. Diapause intensity, metabolic rate trajectories and lipid profiles of directly developing and diapausing animals were studied using pupae and adults of Pieris napi butterflies from a population in which endogenous diapause has been well studied. Endogenous diapause was terminated after 3 months and termination required chilling. Metabolic and post-diapause development rates increased with diapause duration, while the metabolic cost of post-diapause development decreased, indicating that once diapause is terminated, development proceeds at a low rate even at low temperature. Diapausing pupae had larger lipid stores than the directly developing pupae, and lipids constituted the primary energy source during diapause. However, during diapause, lipid stores did not decrease. Thus, despite lipid catabolism meeting the low energy costs of the diapausing pupae, primary lipid store utilization did not occur until the onset of growth and metamorphosis in spring. In line with this finding, diapausing pupae contained low amounts of mitochondria-derived cardiolipins, which suggests a low capacity for fatty acid β-oxidation. While ontogenic development had a large effect on lipid and fatty acid profiles, only small changes in these were seen during diapause. The data therefore indicate that the diapause lipidomic phenotype is developed early, when pupae are still at high temperature, and retained until post-diapause development. PMID: 27445351 [PubMed - indexed for MEDLINE]

Conventional and accelerated-solvent extractions of green tea (camellia sinensis) for metabolomics-based chemometrics.

Wed, 09/08/2017 - 13:31
Conventional and accelerated-solvent extractions of green tea (camellia sinensis) for metabolomics-based chemometrics. J Pharm Biomed Anal. 2017 Jul 29;145:604-610 Authors: Kellogg JJ, Wallace ED, Graf TN, Oberlies NH, Cech NB Abstract Metabolomics has emerged as an important analytical technique for multiple applications. The value of information obtained from metabolomics analysis depends on the degree to which the entire metabolome is present and the reliability of sample treatment to ensure reproducibility across the study. The purpose of this study was to compare methods of preparing complex botanical extract samples prior to metabolomics profiling. Two extraction methodologies, accelerated solvent extraction and a conventional solvent maceration, were compared using commercial green tea [Camellia sinensis (L.) Kuntze (Theaceae)] products as a test case. The accelerated solvent protocol was first evaluated to ascertain critical factors influencing extraction using a D-optimal experimental design study. The accelerated solvent and conventional extraction methods yielded similar metabolite profiles for the green tea samples studied. The accelerated solvent extraction yielded higher total amounts of extracted catechins, was more reproducible, and required less active bench time to prepare the samples. This study demonstrates the effectiveness of accelerated solvent as an efficient methodology for metabolomics studies. PMID: 28787673 [PubMed - as supplied by publisher]

MetExtract II: A software suite for stable isotope assisted untargeted metabolomics.

Wed, 09/08/2017 - 13:31
MetExtract II: A software suite for stable isotope assisted untargeted metabolomics. Anal Chem. 2017 Aug 08;: Authors: Bueschl C, Kluger B, Neumann NKN, Doppler M, Maschietto V, Thallinger GG, Meng-Reiterer J, Krska R, Schuhmacher R Abstract Stable isotope labeling (SIL) techniques have the potential to enhance different aspects of LC-HRMS based, untargeted metabolomics methods including metabolite detection, annotation of unknown metabolites and comparative quantification. In this work, we present MetExtract II, a software toolbox for the detection of biologically derived compounds. It exploits SIL-specific isotope patterns and elution profiles in LC-HRMS(/MS) data. The toolbox consists of three complementary modules (M1-M3): M1 (AllExtract) uses mixtures of uniformly, highly isotope-enriched and native biological samples for the selective detection of the entire accessible metabolome. M2 (TracExtract) is particularly suited to probe the metabolism of endogenous or exogenous secondary metabolites and facilitates the untargeted screening of tracer-derivatives from concurrently metabolized native and uniformly labeled tracer substances. With M3 (FragExtract), MS/MS fragments of corresponding native and uniformly labeled ions are evaluated and automatically assigned with putative sum formulas. Generated results can be graphically illustrated and exported as a comprehensive data matrix that contains all detected pairs of native and labeled metabolite ions that can be used for database queries, metabolome-wide internal standardization and statistical analysis. The software, associated documentation and sample datasets are freely available for non-commercial use at http://metabolomics-ifa.boku.ac.at/metextractII. PMID: 28787149 [PubMed - as supplied by publisher]

Overcoming Sample Matrix Effect in Quantitative Blood Metabolomics Using Chemical Isotope Labeling Liquid Chromatography Mass Spectrometry.

Wed, 09/08/2017 - 13:31
Overcoming Sample Matrix Effect in Quantitative Blood Metabolomics Using Chemical Isotope Labeling Liquid Chromatography Mass Spectrometry. Anal Chem. 2017 Aug 08;: Authors: Chen D, Han W, Su X, Li L, Li L Abstract Blood is widely used for discovery metabolomics to search for disease biomarkers. However, blood sample matrix can have a profound effect on metabolome analysis, which can impose an undesirable restriction on the type of blood collection tubes that can be used for blood metabolomics. We investigated the effect of blood sample matrix on metabolome analysis using a high-coverage and quantitative metabolome profiling technique based on differential chemical isotope labeling (CIL) LC-MS. We used (12)C-/(13C)-dansylation LC-MS to perform relative quantification of the amine/phenol submetabolomes of four types of samples (i.e., serum, EDTA-plasma, heparin-plasma and citrate-plasma) collected from healthy individuals and compare their metabolomic results. From the analysis of 80 plasma and serum samples in experimental triplicate, we detected a total of 3651 metabolites with an average of 1818 metabolites per run (n=240). The number of metabolites detected and the precision and accuracy of relative quantification were found to be independent of the sample type. Within each sample type, the metabolome data set could reveal biological variation (e.g., sex separation). Although the relative concentrations of some individual metabolites might be different in the four types of samples, for sex separation, all 66 significant metabolites with larger fold-changes (FC≥2 and p<0.05) found in at least one sample type could be found in the other types of samples with similar or somewhat reduced, but still significant fold-changes. Our results indicate that CIL LC-MS could overcome the sample matrix effect, thereby greatly broadening the scope of blood metabolomics ‒ any blood samples properly collected in routine clinical settings, including those in biobanks originally used for other purposes, can potentially be used for discovery metabolomics. PMID: 28787119 [PubMed - as supplied by publisher]

The steroid metabolome in women with premenstrual dysphoric disorder during GnRH agonist-induced ovarian suppression: effects of estradiol and progesterone addback.

Wed, 09/08/2017 - 13:31
The steroid metabolome in women with premenstrual dysphoric disorder during GnRH agonist-induced ovarian suppression: effects of estradiol and progesterone addback. Transl Psychiatry. 2017 Aug 08;7(8):e1193 Authors: Nguyen TV, Reuter JM, Gaikwad NW, Rotroff DM, Kucera HR, Motsinger-Reif A, Smith CP, Nieman LK, Rubinow DR, Kaddurah-Daouk R, Schmidt PJ Abstract Clinical evidence suggests that symptoms in premenstrual dysphoric disorder (PMDD) reflect abnormal responsivity to ovarian steroids. This differential steroid sensitivity could be underpinned by abnormal processing of the steroid signal. We used a pharmacometabolomics approach in women with prospectively confirmed PMDD (n=15) and controls without menstrual cycle-related affective symptoms (n=15). All were medication-free with normal menstrual cycle lengths. Notably, women with PMDD were required to show hormone sensitivity in an ovarian suppression protocol. Ovarian suppression was induced for 6 months with gonadotropin-releasing hormone (GnRH)-agonist (Lupron); after 3 months all were randomized to 4 weeks of estradiol (E2) or progesterone (P4). After a 2-week washout, a crossover was performed. Liquid chromatography/tandem mass spectrometry measured 49 steroid metabolites in serum. Values were excluded if >40% were below the limit of detectability (n=21). Analyses were performed with Wilcoxon rank-sum tests using false-discovery rate (q<0.2) for multiple comparisons. PMDD and controls had similar basal levels of metabolites during Lupron and P4-derived neurosteroids during Lupron or E2/P4 conditions. Both groups had significant increases in several steroid metabolites compared with the Lupron alone condition after treatment with E2 (that is, estrone-SO4 (q=0.039 and q=0.002, respectively) and estradiol-3-SO4 (q=0.166 and q=0.001, respectively)) and after treatment with P4 (that is, allopregnanolone (q=0.001 for both PMDD and controls), pregnanediol (q=0.077 and q=0.030, respectively) and cortexone (q=0.118 and q=0.157, respectively). Only sulfated steroid metabolites showed significant diagnosis-related differences. During Lupron plus E2 treatment, women with PMDD had a significantly attenuated increase in E2-3-sulfate (q=0.035) compared with control women, and during Lupron plus P4 treatment a decrease in DHEA-sulfate (q=0.07) compared with an increase in controls. Significant effects of E2 addback compared with Lupron were observed in women with PMDD who had significant decreases in DHEA-sulfate (q=0.065) and pregnenolone sulfate (q=0.076), whereas controls had nonsignificant increases (however, these differences did not meet statistical significance for a between diagnosis effect). Alterations of sulfotransferase activity could contribute to the differential steroid sensitivity in PMDD. Importantly, no differences in the formation of P4-derived neurosteroids were observed in this otherwise highly selected sample of women studied under controlled hormone exposures. PMID: 28786978 [PubMed - in process]

Exercise-Induced Alterations in Skeletal Muscle, Heart, Liver, and Serum Metabolome Identified by Non-Targeted Metabolomics Analysis.

Wed, 09/08/2017 - 13:31
Exercise-Induced Alterations in Skeletal Muscle, Heart, Liver, and Serum Metabolome Identified by Non-Targeted Metabolomics Analysis. Metabolites. 2017 Aug 08;7(3): Authors: Starnes JW, Parry TL, O'Neal SK, Bain JR, Muehlbauer MJ, Honcoop A, Ilaiwy A, Christopher PM, Patterson C, Willis MS Abstract BACKGROUND: The metabolic and physiologic responses to exercise are increasingly interesting, given that regular physical activity enhances antioxidant capacity, improves cardiac function, and protects against type 2 diabetes. The metabolic interactions between tissues and the heart illustrate a critical cross-talk we know little about. METHODS: To better understand the metabolic changes induced by exercise, we investigated skeletal muscle (plantaris, soleus), liver, serum, and heart from exercise trained (or sedentary control) animals in an established rat model of exercise-induced aerobic training via non-targeted GC-MS metabolomics. RESULTS: Exercise-induced alterations in metabolites varied across tissues, with the soleus and serum affected the least. The alterations in the plantaris muscle and liver were most alike, with two metabolites increased in each (citric acid/isocitric acid and linoleic acid). Exercise training additionally altered nine other metabolites in the plantaris (C13 hydrocarbon, inosine/adenosine, fructose-6-phosphate, glucose-6-phosphate, 2-aminoadipic acid, heptadecanoic acid, stearic acid, alpha-tocopherol, and oleic acid). In the serum, we identified significantly decreased alpha-tocopherol levels, paralleling the increases identified in plantaris muscle. Eleven unique metabolites were increased in the heart, which were not affected in the other compartments (malic acid, serine, aspartic acid, myoinositol, glutamine, gluconic acid-6-phosphate, glutamic acid, pyrophosphate, campesterol, phosphoric acid, creatinine). These findings complement prior studies using targeted metabolomics approaches to determine the metabolic changes in exercise-trained human skeletal muscle. Specifically, exercise trained vastus lateralus biopsies had significantly increased linoleic acid, oleic acid, and stearic acid compared to the inactive groups, which were significantly increased in plantaris muscle in the present study. CONCLUSIONS: While increases in alpha-tocopherol have not been identified in muscle after exercise to our knowledge, the benefits of vitamin E (alpha-tocopherol) supplementation in attenuating exercise-induced muscle damage has been studied extensively. Skeletal muscle, liver, and the heart have primarily different metabolic changes, with few similar alterations and rare complementary alterations (alpha-tocopherol), which may illustrate the complexity of understanding exercise at the organismal level. PMID: 28786928 [PubMed]

Evaluation of metabolism of azo dyes and their effects on Staphylococcus aureus metabolome.

Wed, 09/08/2017 - 13:31
Related Articles Evaluation of metabolism of azo dyes and their effects on Staphylococcus aureus metabolome. J Ind Microbiol Biotechnol. 2017 Aug 07;: Authors: Sun J, Jin J, Beger RD, Cerniglia CE, Chen H Abstract Dyes containing one or more azo linkages are widely applied in cosmetics, tattooing, food and drinks, pharmaceuticals, printing inks, plastics, leather, as well as paper industries. Previously we reported that bacteria living on human skin have the ability to reduce some azo dyes to aromatic amines, which raises potential safety concerns regarding human dermal exposure to azo dyes such as those in tattoo ink and cosmetic colorant formulations. To comprehensively investigate azo dye-induced toxicity by skin bacteria activation, it is very critical to understand the mechanism of metabolism of the azo dyes at the systems biology level. In this study, an LC/MS-based metabolomics approach was employed to globally investigate metabolism of azo dyes by Staphylococcus aureus as well as their effects on the metabolome of the bacterium. Growth of S. aureus in the presence of Sudan III or Orange II was not affected during the incubation period. Metabolomics results showed that Sudan III was metabolized to 4-(phenyldiazenyl) aniline (48%), 1-[(4-aminophenyl) diazenyl]-2-naphthol (4%) and eicosenoic acid Sudan III (0.9%). These findings indicated that the azo bond close to naphthalene group of Sudan III was preferentially cleaved compared with the other azo bond. The metabolite from Orange II was identified as 4-aminobenzene sulfonic acid (35%). A much higher amount of Orange II (~90×) was detected in the cell pellets from the active viable cells compared with those from boiled cells incubated with the same concentration of Orange II. This finding suggests that Orange II was primarily transported into the S. aureus cells for metabolism, instead of the theory that the azo dye metabolism occurs extracellularly. In addition, the metabolomics results showed that Sudan III affected energy pathways of the S. aureus cells, while Orange II had less noticeable effects on the cells. In summary, this study provided novel information regarding azo dye metabolism by the skin bacterium, the effects of azo dyes on the bacterial cells and the important role on the toxicity and/or inactivation of these compounds due to microbial metabolism. PMID: 28786013 [PubMed - as supplied by publisher]

Stress-Related Mitogen-Activated Protein Kinases Stimulate the Accumulation of Small Molecules and Proteins in Arabidopsis thaliana Root Exudates.

Wed, 09/08/2017 - 13:31
Related Articles Stress-Related Mitogen-Activated Protein Kinases Stimulate the Accumulation of Small Molecules and Proteins in Arabidopsis thaliana Root Exudates. Front Plant Sci. 2017;8:1292 Authors: Strehmel N, Hoehenwarter W, Mönchgesang S, Majovsky P, Krüger S, Scheel D, Lee J Abstract A delicate balance in cellular signaling is required for plants to respond to microorganisms or to changes in their environment. Mitogen-activated protein kinase (MAPK) cascades are one of the signaling modules that mediate transduction of extracellular microbial signals into appropriate cellular responses. Here, we employ a transgenic system that simulates activation of two pathogen/stress-responsive MAPKs to study release of metabolites and proteins into root exudates. The premise is based on our previous proteomics study that suggests upregulation of secretory processes in this transgenic system. An advantage of this experimental set-up is the direct focus on MAPK-regulated processes without the confounding complications of other signaling pathways activated by exposure to microbes or microbial molecules. Using non-targeted metabolomics and proteomics studies, we show that MAPK activation can indeed drive the appearance of dipeptides, defense-related metabolites and proteins in root apoplastic fluid. However, the relative levels of other compounds in the exudates were decreased. This points to a bidirectional control of metabolite and protein release into the apoplast. The putative roles for some of the identified apoplastic metabolites and proteins are discussed with respect to possible antimicrobial/defense or allelopathic properties. Overall, our findings demonstrate that sustained activation of MAPKs alters the composition of apoplastic root metabolites and proteins, presumably to influence the plant-microbe interactions in the rhizosphere. The reported metabolomics and proteomics data are available via Metabolights (Identifier: MTBLS441) and ProteomeXchange (Identifier: PXD006328), respectively. PMID: 28785276 [PubMed]

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