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

The challenges of developing and optimising an assay to measure 25-hydroxyvitamin D in saliva.

Mon, 29/07/2019 - 14:01
Related Articles The challenges of developing and optimising an assay to measure 25-hydroxyvitamin D in saliva. J Steroid Biochem Mol Biol. 2019 Jul 25;:105437 Authors: Clarke MW, Black LJ, Hart PH, Jones AP, Palmer DJ, Siafarikas A, Lucas RM, Gorman S Abstract Accurately detecting vitamin D deficiency (defined as concentration in blood of 25-hydroxyvitamin D (25(OH)D), <20 ng/mL) is important for both clinicians and researchers. Drawing blood may be difficult in some populations, such as infants and children. We thus explored the development of a method to measure 25(OH)D concentrations in saliva, using a liquid chromatography tandem mass spectrometry (LC-MS/MS) assay. Using 25(OH)D3 standards spiked into synthetic saliva, we generated a standard curve with high correlation (r = 0.999, Pearson's); the intra-assay and inter-assay variation were ≤3.2% and ≤13.2% (CV%), respectively. Passive collection of saliva via drooling into glass or polypropylene tubes yielded higher levels of 25(OH)D3 than chewing on a synthetic swab. Chewing gum for at least 4 minutes reduced saliva levels of 25(OH)D3. Differences in the levels of 25(OH)D3 in saliva between the passive drooling and stimulated swab-chewing methods were normalised by adjusting for measured levels of vitamin D binding protein in saliva. Freezing samples immediately, or after 24 h of refrigeration did not affect 25(OH)D3 levels. When saliva levels of 25(OH)D3 were averaged from samples collected daily for three consecutive days, for which an additional centrifugation step was performed after samples were defrosted (to remove mucin), there was a positive (but non-significant) correlation between 25(OH)D3 levels in saliva and serum (r = 0.57, p = 0.24, Pearson's) with significant correlations (r ≥ 0.88, p < 0.05) observed after further adjusting for saliva flow rate. The time of day of the collection made little difference to 25(OH)D3 levels measured in saliva. In conclusion, we have developed an LC-MS/MS assay that accurately measures saliva 25(OH)D3 levels, which correlated with serum levels. However, for a measurement that correlates with serum 25(OH)D it may be necessary to average results from saliva collected on three consecutive days, and adjust for differences in saliva flow rate. This would increase costs, and combined with the processing requirements for samples, could limit the applicability of this assay to large cohort and field studies. PMID: 31352025 [PubMed - as supplied by publisher]

The Future Of Omics For Clinical Practice.

Mon, 29/07/2019 - 14:01
Related Articles The Future Of Omics For Clinical Practice. Ann Allergy Asthma Immunol. 2019 Jul 25;: Authors: Long A, Bunning B, Borro M, Sampath V, Nadeau KC Abstract Dr. Kari Nadeau: Dr. Nadeau receives grant support from NIAID, Food Allergy Research & Education (FARE), End Allergies Together, Allergenis, Ukko; personal Fees from Regeneron, AstraZeneca, ImmuneWorks, Cour; sponsored research from Novartis, Sanofi, Astellas, Nestle; Sponsored research for clinical trials from Genentech, Aimmune Therapeutics, DBV Technologies, AnaptysBio, Stallergenes-Greer, Regeneron, and Adare Pharmaceuticals; Data and Safety Monitoring Board member at Novartis and NHLBI; co-founded Before Brands, Alladapt Immunotherapeutics, and ForTra; grant awardee for NIAID, NHLBI, NIEHS, and EPA; Director for FARE and World Allergy Organization (WAO) Center of Excellence. All other authors declare no conflict of interest. PMID: 31351978 [PubMed - as supplied by publisher]

metabolomics; +44 new citations

Sun, 28/07/2019 - 13:54
44 new pubmed citations were retrieved for your search. Click on the search hyperlink below to display the complete search results: metabolomics These pubmed results were generated on 2019/07/28PubMed comprises more than millions of citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.

metabolomics; +29 new citations

Fri, 26/07/2019 - 13:35
29 new pubmed citations were retrieved for your search. Click on the search hyperlink below to display the complete search results: metabolomics These pubmed results were generated on 2019/07/26PubMed comprises more than millions of citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.

metabolomics; +46 new citations

Thu, 25/07/2019 - 13:26
46 new pubmed citations were retrieved for your search. Click on the search hyperlink below to display the complete search results: metabolomics These pubmed results were generated on 2019/07/25PubMed comprises more than millions of citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.

metabolomics; +22 new citations

Wed, 24/07/2019 - 16:21
22 new pubmed citations were retrieved for your search. Click on the search hyperlink below to display the complete search results: metabolomics These pubmed results were generated on 2019/07/24PubMed comprises more than millions of citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.

metabolomics; +18 new citations

Tue, 23/07/2019 - 16:09
18 new pubmed citations were retrieved for your search. Click on the search hyperlink below to display the complete search results: metabolomics These pubmed results were generated on 2019/07/23PubMed comprises more than millions of citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.

metabolomics; +18 new citations

Tue, 23/07/2019 - 13:08
18 new pubmed citations were retrieved for your search. Click on the search hyperlink below to display the complete search results: metabolomics These pubmed results were generated on 2019/07/23PubMed comprises more than millions of citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.

metabolomics; +16 new citations

Mon, 22/07/2019 - 15:56
16 new pubmed citations were retrieved for your search. Click on the search hyperlink below to display the complete search results: metabolomics These pubmed results were generated on 2019/07/22PubMed comprises more than millions of citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.

metabolomics; +16 new citations

Mon, 22/07/2019 - 12:53
16 new pubmed citations were retrieved for your search. Click on the search hyperlink below to display the complete search results: metabolomics These pubmed results were generated on 2019/07/22PubMed comprises more than millions of citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.

Serum metabolomics profiling and potential biomarkers of myopia using LC-QTOF/MS.

Sun, 21/07/2019 - 18:44
Serum metabolomics profiling and potential biomarkers of myopia using LC-QTOF/MS. Exp Eye Res. 2019 Jul 17;:107737 Authors: Dai L, Yang W, Qin X, Li Y, Cao H, Zhou C, Wang Y Abstract Myopia is the most common form of refractive eye disease, and the prevalence is increasing rapidly worldwide. However, the key metabolic alterations in individuals with high myopia are not understood clearly, and serum biomarkers remain to be determined. The objectives of this study were to identify serum biomarkers and investigate the metabolic alterations of myopia. The serum metabolomics profiling was investigated on 30 high myopia cases and 30 controls (without myopia) using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF/MS), and an independent additional cohort including 20 cases and 19 controls were investigated to validate potential metabolite candidates for biomarkers. According to the metabolic differences, the myopia patients and controls could be divided into different clusters and nine metabolites were found to be closely correlated with myopia. In the cohort of validation, eight metabolites were confirmed. Metabolic pathway analyses of these metabolites of high myopia involved abnormal phospholipid, diacylglycerol, amino acid, and vitamin metabolism, which were closely correlated with oxidative stress and inflammation. Multiple logistic regression analyses showed that γ-glutamyltyrosine and 12-oxo-20-trihydroxy-leukotriene B4 were potential biomarkers of myopia with a combined high sensitivity (97%), specificity (90%), and area under the curve value (0.983). These findings may contribute to an understanding of the pathophysiological changes and pathogenesis of myopia, and provide novel insight into the early prevention and control of high myopia. PMID: 31325450 [PubMed - as supplied by publisher]

Identification of coronary heart disease biomarkers with different severities of coronary stenosis in human urine using non-targeted metabolomics based on UPLC-Q-TOF/MS.

Sun, 21/07/2019 - 18:44
Identification of coronary heart disease biomarkers with different severities of coronary stenosis in human urine using non-targeted metabolomics based on UPLC-Q-TOF/MS. Clin Chim Acta. 2019 Jul 17;: Authors: Huang M, Zhao H, Gao S, Liu Y, Liu Y, Zhang T, Cai X, Li Z, Li L, Li Y, Yu C Abstract BACKGROUND: Coronary heart disease (CHD) is the leading cause of death worldwide, and its pathogenesis has attracted much attention. Metabolomics serves as an important tool for diagnosing diseases and exploring their pathogenesis in recent years. In this study, CHD patients were studied by comparing them with normal subjects to elucidate biomarkers that are linearly correlated with the severity of coronary stenosis. METHODS: High-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was used to analyze the urine metabolites of CHD patients and normal subjects. A total of 131 subjects included 23 patients who presented with 50-69% coronary stenosis, 22 with 70-89% stenosis, 29 with 90-99% stenosis, 24 with 100% stenosis, and 29 normal subjects. RESULTS: A total of 15 potential biomarkers associated with CHD were identified, and among them 5 biomarkers were linearly correlated with the severity of coronary stenosis in CHD patients. The metabolic pathways involved were amino acid metabolism, fatty acid metabolism, energy metabolism, and nucleotide metabolism. CONCLUSION: This study identified the biomarkers and metabolic pathways that may be involved in the occurrence and development of CHD, laying a theoretical foundation for better diagnosis and treatment of CHD in the future. PMID: 31325445 [PubMed - as supplied by publisher]

Metabolomics Analysis of Nutrient Metabolism in β-Cells.

Sun, 21/07/2019 - 18:44
Metabolomics Analysis of Nutrient Metabolism in β-Cells. J Mol Biol. 2019 Jul 17;: Authors: Spégel P, Mulder H Abstract The islets of Langerhans harbor multiple endocrine cell-types that continuously respond to circulating nutrient levels in order to adjust their secretion of catabolic and anabolic hormones. Stimulus-secretion coupling in these cells is largely of metabolic nature, i.e. metabolism of nutrient fuels yields signals that trigger and amplify secretion of hormones. Hence, metabolism in this micro-organ is in a major way in control of whole-body metabolism. Therefore, insights into islet metabolism is critical to understand how secretion of insulin is regulated and why it is perturbed in type 2 diabetes. Metabolomics aims at characterizing a wide spectrum of metabolites in cells, tissues and body fluids. For this reason, this technique is well-suited to supply information on stimulus-secretion coupling. Here, we summarize metabolomics studies in islets and β-cells, highlight important discoveries that would have been difficult to make without this technology but also raise awareness of challenges and bottlenecks that curtail its use in metabolic research. PMID: 31325441 [PubMed - as supplied by publisher]

Metabolite variation in three edible Italian Allium cepa L. by NMR-based metabolomics: a comparative study in fresh and stored bulbs.

Sun, 21/07/2019 - 18:44
Related Articles Metabolite variation in three edible Italian Allium cepa L. by NMR-based metabolomics: a comparative study in fresh and stored bulbs. Metabolomics. 2019 Jul 19;15(8):105 Authors: Saviano G, Paris D, Melck D, Fantasma F, Motta A, Iorizzi M Abstract INTRODUCTION: In fruits and vegetables, comparative analysis of metabolic plant profiles has a high potential for quality control of active components. Onion (Allium cepa L.) is used fresh or stored as food, spice, and in traditional medicine. Its metabolic content, often with nutraceutical value, makes its level an important factor in agronomic production. OBJECTIVE: To describe for the first time the metabolome of "San Pietro" white onion (WP), and compare its chemical profile with the red onion var. Tropea (RT) and the yellow onion var. Montoro (CM). Furthermore, we also aim to obtain a multivariate model based on NMR fingerprints to discriminate the three Italian A. cepa L. cultivars. METHODS: For the chemical fingerprinting we used NMR-based metabolomics. We investigated the aqueous and chloroform extracts of fresh onion at harvesting time, and after 9-month storage. Principal component analysis (PCA), Partial least squares discriminant analysis (PLS-DA) and Orthogonal partial least squares (OPLS-DA) were used to build reliable models. RESULTS: We obtained a clear discrimination of A. cepa L. varieties for the fresh and stored batches. The statistical model highlighted higher levels of fructo-oligosaccharides (FOS) in the fresh WP; RT showed a high content of glucose, citrate and amino acids, while CM had many sulfur components. In the stored samples (CMS, RTS), carbohydrates and sulfur components decreased, while in WPS the free monosaccharides concentration increased. Linoleic acid was overexpressed in the apolar extracts of CMF and WPF cultivars. CONCLUSION: Metabolomics allows a reliable differentiation among onion varieties, and highlights the potential of fingerprinting for food authentication purposes. PMID: 31325058 [PubMed - in process]

Human milk and infant formula differentially alters the microbiota composition and functional gene relative abundance in the small and large intestines in weanling rats.

Sun, 21/07/2019 - 18:44
Related Articles Human milk and infant formula differentially alters the microbiota composition and functional gene relative abundance in the small and large intestines in weanling rats. Eur J Nutr. 2019 Jul 19;: Authors: Liu Z, Subbaraj A, Fraser K, Jia H, Chen W, Day L, Roy NC, Young W Abstract PURPOSE: Human breast milk is the optimal source of nutrients for growing infants. However, many circumstances can arise which preclude breast milk feeding, leading to the use of infant formula, including during the weaning period. Many diet-related effects are modulated by the gut microbiome. Therefore, we investigated the effect of human milk (HM) or infant formula (IF) on the gut microbiota in weanling rats. METHODS: The gut microbiota of weanling male Sprague-Dawley rats fed HM or IF for 28 days was analysed by shotgun metagenome sequencing. Caecal contents were analysed by liquid chromatography-mass spectrometry metabolomics. RESULTS: Numerous genera within the Proteobacteria phylum were relatively more abundant in the ileum, caecum, and colon of rats fed HM, including ileal Escherichia (HM = 9.6% ± 4.3 SEM; IF = 0.9% ± 0.3 SEM; P = 0.03). Other taxa that differed between HM- and IF-fed rats included Prevotella and Ruminococcus. Overall, more differences were observed in the ileum than the caecum and colon between rats fed HM and IF. For the rats fed IF, in the ileum, the relative abundance of Bifidobacterium was higher (HM = 1.7% ± 0.7 SEM; IF = 5.0% ± 1.5 SEM; P = 0.04) with gene functions related to carbohydrate and amino acid metabolism also decreased. In the caecum, metabolic features such as bile acids were elevated while amino sugars were also decreased. CONCLUSION: Our results show that HM and IF composition differences are reflected in the gut microbiome composition and function in both the small and large intestines. PMID: 31325042 [PubMed - as supplied by publisher]

Acute and short-term administrations of delta-9-tetrahydrocannabinol modulate major gut metabolomic regulatory pathways in C57BL/6 mice.

Sun, 21/07/2019 - 18:44
Related Articles Acute and short-term administrations of delta-9-tetrahydrocannabinol modulate major gut metabolomic regulatory pathways in C57BL/6 mice. Sci Rep. 2019 Jul 19;9(1):10520 Authors: Oza M, Becker W, Gummadidala PM, Dias T, Omebeyinje MH, Chen L, Mitra C, Jesmin R, Chakraborty P, Sajish M, Hofseth LJ, Banerjee K, Wang Q, Moeller PDR, Nagarkatti M, Nagarkatti P, Chanda A Abstract Delta-9-tetrahydrocannabinol (THC) is the primary psychoactive compound in Cannabis, which is studied extensively for its medicinal value. A central gap in the science is the underlying mechanisms surrounding THC's therapeutic effects and the role of gut metabolite profiles. Using a mass-spectrometry based metabolomics, we show here that intraperitoneal injection of THC in C57BL/6 mice modulates metabolic profiles that have previously been identified as integral to health. Specifically, we investigated the effects of acute (single THC injection denoted here as '1X') and short -term (five THC injections on alternate days denoted as '5X') THC administration on fecal and intestinal tissue metabolite profiles. Results are consistent with the hypothesis that THC administration alters host metabolism by targeting two prominent lipid metabolism pathways: glycerophospholipid metabolism and fatty acid biosynthesis. PMID: 31324830 [PubMed - in process]

Boosting NAD+ with a small molecule that activates NAMPT.

Sun, 21/07/2019 - 18:44
Related Articles Boosting NAD+ with a small molecule that activates NAMPT. Nat Commun. 2019 Jul 19;10(1):3241 Authors: Gardell SJ, Hopf M, Khan A, Dispagna M, Hampton Sessions E, Falter R, Kapoor N, Brooks J, Culver J, Petucci C, Ma CT, Cohen SE, Tanaka J, Burgos ES, Hirschi JS, Smith SR, Sergienko E, Pinkerton AB Abstract Pharmacological strategies that boost intracellular NAD+ are highly coveted for their therapeutic potential. One approach is activation of nicotinamide phosphoribosyltransferase (NAMPT) to increase production of nicotinamide mononucleotide (NMN), the predominant NAD+ precursor in mammalian cells. A high-throughput screen for NAMPT activators and hit-to-lead campaign yielded SBI-797812, a compound that is structurally similar to active-site directed NAMPT inhibitors and blocks binding of these inhibitors to NAMPT. SBI-797812 shifts the NAMPT reaction equilibrium towards NMN formation, increases NAMPT affinity for ATP, stabilizes phosphorylated NAMPT at His247, promotes consumption of the pyrophosphate by-product, and blunts feedback inhibition by NAD+. These effects of SBI-797812 turn NAMPT into a "super catalyst" that more efficiently generates NMN. Treatment of cultured cells with SBI-797812 increases intracellular NMN and NAD+. Dosing of mice with SBI-797812 elevates liver NAD+. Small molecule NAMPT activators such as SBI-797812 are a pioneering approach to raise intracellular NAD+ and realize its associated salutary effects. PMID: 31324777 [PubMed - in process]

l-Proline protects mice challenged by Klebsiella pneumoniae bacteremia.

Sun, 21/07/2019 - 18:44
Related Articles l-Proline protects mice challenged by Klebsiella pneumoniae bacteremia. J Microbiol Immunol Infect. 2019 Jul 05;: Authors: Chen X, Qin S, Zhao X, Zhou S Abstract OBJECTIVE: K. pneumoniae, a common pathogen that frequently causes bacteremia in clinic, is unresponsive to most of known antibiotics, thus cumulatively exacerbating empirical therapy failures. Effective strategies to control Klebsiella pneumoniae bacteremia are in high demand. One possibility is to mobilize host defense mechanisms against bacterial pathogens. METHODS: We employed GC/MS-based metabolomics to identify the changes of metabolism in mice challenged by K. pneumoniae (ATCC 43816) bacteremia. RESULTS: Compared with the mice that compromised from K. pneumoniae bacteremia, mice that survived from infection displayed the varied metabolomic profile. The differential analysis of metabolome showed that Ethanedioic acid, d-Glucose, l-Glutamine, Myo-inositol, and l-Proline were more likely associated with the host surviving a K. pneumoniae bacteremia. Further pathway enrichment analysis proposed that arginine and proline metabolism involved in outcome of K. pneumoniae bacteremia. The follow-up data showed that exogenous l-Proline but not d-Proline could decline the loads of Klebsiella pneumonia in infected blood and tissues (lung, liver and spleen) and increase the mouse survival. CONCLUSION: Our study provides an exercisable strategy of identifying metabolic biomarkers from surviving host and highlights the possibility of utilizing the metabolic biomarker as a therapy for K. pneumoniae bacteremia. PMID: 31324551 [PubMed - as supplied by publisher]

Biomarkers in neonatal hypoxic-ischemic encephalopathy-Review of the literature to date and future directions for research.

Sun, 21/07/2019 - 18:44
Related Articles Biomarkers in neonatal hypoxic-ischemic encephalopathy-Review of the literature to date and future directions for research. Handb Clin Neurol. 2019;162:281-293 Authors: Murray DM Abstract The widespread introduction of therapeutic hypothermia as a standard of care in hypoxic-ischemic encephalopathy (HIE) has brought increasing pressure on clinicians to make an early and accurate assessment of the degree of hypoxic injury (HI) that has occurred and the severity of the encephalopathy that will ensue. No single blood-based marker is currently robust enough to detect significant HI or predict outcome. However, research in the field has been active in the last 10 years and we know that HIE is associated with predictable alterations in the expression of a number of inflammatory proteins, neuron-specific proteins, metabolite pathways, and microRNA. These alterations evolve quickly over the first hours and days of life. Predictive power varies depending on the timing of measurement of the biomarker, the sample type, and the case mix of the cohort examined. Combining clinical data with biochemical measurements is currently the most likely path toward improved detection and prediction of outcome in neonatal HIE. PMID: 31324315 [PubMed - in process]

A highly sensitive ultra-high performance liquid chromatography/tandem mass spectrometry method with in-source fragmentation for rapid quantification of raspberry ketone.

Sun, 21/07/2019 - 18:44
Related Articles A highly sensitive ultra-high performance liquid chromatography/tandem mass spectrometry method with in-source fragmentation for rapid quantification of raspberry ketone. J Food Drug Anal. 2019 Jul;27(3):778-785 Authors: Yuan B, Zhao D, Du R, Kshatriya D, Bello NT, Simon JE, Wu Q Abstract Raspberry ketone (RK) is the characteristic aromatic compound in raspberry (Rubus idaeus L.) with wide applications as food additive and anti-obesity agent. However, quantification of RK has presented difficulties in MS detection and reliable LC-MS method for RK analysis in literature is in limit to date. In order to facilitate quality control of raspberry derived products and RK metabolomics study, this study aimed to develop a validated and sensitive UHPLC-MS/MS method. Strong in-source fragmentation was noted and the fragmental ion of 107 m/z produced was selected as the precursor ion for MRM detection, and as such the electrospray ionization performance was optimized by fractional factorial design to accommodate such ion-source dissociation behavior as well as its moderate volatility. A pathway involving the formation of quinone-like structure with strong conjugation was proposed to explain the intense in-source fragmentation. The MRM transition was optimized with product ion of 77 m/z selected as the quantifier ion. The method featured low limit of quantification of ∼2 ng/mL and allowed for rapid detection of RK in fresh raspberries following direct sample preparation. RK contents were found to be higher from locally grown and harvested farm sources compared to commercial products shipped into the state, and higher in those at late-stage compared with early-stage maturity. No correlations in RK content between organic and non-organic labels were noted. PMID: 31324293 [PubMed - in process]

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