The ABRF Metabolomics Research Group 2013 Study: Investigation of Spiked Compound Differences in a Human Plasma Matrix

Metabolomics is an emerging field that involves qualitative and quantitative measurements of small molecule metabolites in a biological system. These measurements can be useful for developing biomarkers for diagnosis, prognosis, or predicting response to therapy. Currently, a wide variety of metabolomics approaches, including nontargeted and targeted profiling, are used across laboratories on a routine basis. A diverse set of analytical platforms, such as NMR, gas chromatography-mass spectrometry, Orbitrap mass spectrometry, and time-of-flight-mass spectrometry, which use various chromatographic and ionization techniques, are used for resolution, detection, identification, and quantitation of metabolites from various biological matrices. However, few attempts have been made to standardize experimental methodologies or comparative analyses across different laboratories. The Metabolomics Research Group of the Association of Biomolecular Resource Facilities organized a “round-robin” experiment type of interlaboratory study, wherein human plasma samples were spiked with different amounts of metabolite standards in 2 groups of biologic samples (A and B). The goal was a study that resembles a typical metabolomics analysis. Here, we report our efforts and discuss challenges that create bottlenecks for the field. Finally, we discuss benchmarks that could be used by laboratories to compare their methodologies.     

Seven New Sesquiterpenoids from the Fruits of Schisandra sphenanthera

Fractionation of the EtOH extract from the fruits of Schisandra sphenanthera resulted in the isolation of seven new sesquiterpenoids, 1 – 7 , in addition to the known metabolites 8 – 23 . Among them, schiscupatetralin A ( 1 ) possesses an unprecedented structure with a C? C bond between cuparenol and tetralin. The isolated new compounds were evaluated for their anti‐HSV‐1 and anti‐inflammatory activities. The results revealed that compound 4 exhibited anti‐HSV‐1 activity, while compound 6 showed a significant anti‐inflammatory activity.     

Metabolomics: building on a century of biochemistry to guide human health

Medical diagnosis and treatment efficacy will improve significantly when a more personalized system for health assessment is implemented. This system will require diagnostics that provide sufficiently detailed information about the metabolic status of individuals such that assay results will be able to guide food, drug and lifestyle choices to maintain or improve distinct aspects of health without compromising others. Achieving this goal will use the new science of metabolomics – comprehensive metabolic profiling of individuals linked to the biological understanding of human integrative metabolism. Candidate technologies to accomplish this goal are largely available, yet they have not been brought into practice for this purpose. Metabolomic technologies must be sufficiently rapid, accurate and affordable to be routinely accessible to both healthy and acutely ill individuals. The use of metabolomic data to predict the health trajectories of individuals will require bioinformatic tools and quantitative reference databases. These databases containing metabolite profiles from the population must be built, stored and indexed according to metabolic and health status. Building and annotating these databases with the knowledge to predict how a specific metabolic pattern from an individual can be adjusted with diet, drugs and lifestyle to improve health represents a logical application of the biochemistry knowledge that the life sciences have produced over the past 100 years.     

Application of analytical and preparative high-speed counter-current chromatography for the separation of Z-ligustilide from a crude extract of Angelica sinensis

Z-Ligustilide was separated and purified from the traditional Chinese medicinal plant Angelica sinensis by high-speed counter-current chromatography (HSCCC). Analytical HSCCC was first used for the systematic selection of the two-phase solvent system. Preparative HSCCC separation was performed with a two-phase solvent system composed of petroleum ether (60-90 degrees C)-ethanol-water at an optimum volume ratio of 10:17:10 (v/v). A total of 38 mg Z-ligustilide at 98.8% purity was obtained in one step from 200 mg crude extract as determined by HPLC analysis. The structure of the target compound was identified by electron impact ionisation mass spectrometry.     

Metabolomics should be deployed in the identification and characterization of gene‐edited crops

Gene‐editing techniques are currently revolutionizing biology, allowing far greater precision than previous mutagenic and transgenic approaches. They are becoming applicable to a wide range of plant species and biological processes. Gene editing can rapidly improve a range of crop traits, including disease resistance, abiotic stress tolerance, yield, nutritional quality and additional consumer traits. Unlike transgenic approaches, however, it is not facile to forensically detect gene‐editing events at the molecular level, as no foreign DNA exists in the elite line. These limitations in molecular detection approaches are likely to focus more attention on the products generated from the technology than on the process in itself. Rapid advances in sequencing and genome assembly increasingly facilitate genome sequencing as a means of characterizing new varieties generated by gene‐editing techniques. Nevertheless, subtle edits such as single base changes or small deletions may be difficult to distinguish from normal variation within a genotype. Given these emerging scenarios, downstream ‘omics’ technologies reflective of edited affects, such as metabolomics, need to be used in a more prominent manner to fully assess compositional changes in novel foodstuffs. To achieve this goal, metabolomics or ‘non‐targeted metabolite analysis’ needs to make significant advances to deliver greater representation across the metabolome. With the emergence of new edited crop varieties, we advocate: (i) concerted efforts in the advancement of ‘omics’ technologies, such as metabolomics, and (ii) an effort to redress the use of the technology in the regulatory assessment for metabolically engineered biotech crops.     

人工养殖的扬子鳄适应自然环境的能力

为了复壮扬子鳄野生种群数量,实施养殖扬子鳄放归自然是重要途径之一.通过对人工养殖扬子鳄进行野外放归的试验研究,以探明其在野外环境下活动情况和适应能力.2007年6～11月,采用无线电遥测跟踪、望远镜观察和夜间辅助灯光望远镜观察等方法,对健康的4条5.5年龄和3条2.5年龄的人工养殖扬子鳄在自然环境下活动情况进行观察;在2008和2009年的5月,将这些鳄重捕后测量它们的大小并察看生长情况.结果显示:扬子鳄的释放初期经历了过渡期和争夺领域期2个阶段,20天后处于不同的区域内,具领域特征和营建洞穴行为,能够安全越冬;逐渐恢复野生习性,自我保护的警惕性提高,逃避威胁的能力加强;小龄鳄生长明显,大龄鳄没有明显生长,这与它们所处环境的食物种类有关;鳄性成熟后,能够成功繁殖.总之,人工养殖扬子鳄能够适应相应的自然环境,能够正常生活、生存和繁衍.     

寄主植物对荔枝蒂蛀虫产卵的引诱作用

试验观察荔枝蒂蛀虫Conopomorpha sinensisBradley对寄主植物品种、器官及器官部位的产卵选择性规律。结果表明,与老梢相比,荔枝蒂蛀虫明显倾向于选择嫩梢产卵;4个荔枝品种果枝中以妃子笑上落卵数最多,在淮枝、糯米糍荔枝果实上较少,桂味荔枝上最少;不同的荔枝品种果实气味引诱雌虫产卵的能力不同,其中妃子笑引诱力最强,淮枝、糯米糍引诱力较低,桂味最低;在同一荔枝果实不同部位的落卵量,以果皮上、下两部分较大,全果明显低于果皮上部,而荔枝肉和荔枝核上没有落卵。     

Synthesis,crystal structure,optical properties and antibacterial evaluation of novel imidazo[1, 5‐a]pyridine derivatives bearing a hydrazone moiety

A series of novel imidazo[1,5‐a]pyridine‐hydrazone derivatives were synthesized and characterized by infrared spectroscopy (IR), ¹H NMR, ¹³C NMR and high resolution mass spectrometer (HRMS). Typically, the spatial structure of compound 3j was determined using X‐ray diffraction analysis. The UV–vis absorption and fluorescence spectral characteristics of the compounds in dichloromethane and acetonitrile were investigated. Absorption peaks could be observed in the wavelength range 290–450 nm. It can also be seen that they display very similar maximum emission. The group attached to hydrazone hardly influenced the maximum emission. Furthermore, all the compounds were evaluated for antibacterial activity and were found to be more effective against Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa and Shigella compared with chloramphenicol. Copyright © 2013 John Wiley & Sons, Ltd.     

食果鸟白头鹎与接骨草的分布

作为种子的传播者 ,以果实为生的动物在所食植物的繁殖周期中起着关键作用。尽管动植物相互作用的研究自达尔文开始就引起了进化生物学家和生态学家的关注 ,但和传粉者与植物的相互作用的研究相比 ,种子传播者与植物相互作用的研究在 2 0世纪 70年代后才真正开始[1] 。植物学家     

Application of a sensitive near‐field microwave microprobe to the nondestructive characterization of microbial rhodopsin

We study the opto‐electrical properties of Natronomonas pharaonis sensory rhodopsin II (NpSRII) by using a near‐field microwave microprobe (NFMM) under external light illumination. To investigate the possibility of application of NFMM to biological macromolecules, we used time dependent properties of NPSRII before/after light activation which has three distinct states – ground‐state, M‐state, and O‐state. The diagnostic ability of NFMM is demonstrated by measuring the microwave reflection coefficient (S₁₁) spectrum of NpSRII under steady‐state illumination in the wavelength range of 350–650 nm. Moreover, we present microwave reflection coefficient S₁₁ spectra in the same wavelength range for two fast‐photocycling rhodopsins: green light‐absorbing proteorhodopsin (GPR) and Gloeobacter rhodopsin (GR). In addition the frequency sweep shift can be detected completely even for tiny amounts of sample (～10^–3 OD of rhodopsin). Based on these results NFMM shows both very high sensitivity for detecting conformational changes and produces a good time‐resolved spectrum. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Metabolomic evaluation of rat liver and testis to characterize the toxicity of triazole fungicides

The effects of two triazole fungicides, myclobutanil and triadimefon, on endogenous rat metabolite profiles in blood serum, liver, and testis was assessed using proton nuclear magnetic resonance (¹H-NMR) spectroscopy. Adult male Sprague-Dawley rats were dosed daily by gavage for 14 days with myclobutanil or triadimefon, at two dose levels for each triazole. Following exposure, serum, liver, and testis were collected and processed for NMR analysis. Principal components analysis (PCA) and partial least squares discriminant analysis (PLS-DA) of the resulting spectra were used to determine changes in metabolite profiles as a result of exposure. Using this approach, responses common to both triazoles were identified, as well as responses indicative of differences in the toxicity of these two compounds. Although changes were observed in serum metabolites following exposure, none were robust enough to be considered a biomarker of exposure/effect. A number of metabolic changes were, however, observed in the liver with both triazoles, particularly in metabolites related to the methionine cycle. The testes of myclobutanil-exposed animals displayed altered levels of creatine and creatinine, consistent with testicular toxicity. Overall, the results of this study support the possible application of a metabolomics approach to assessing the toxicity of triazole fungicides and identifying biomarkers of exposure and/or effect.RNM supported by EPA/NCSU Cooperative Training Agreement #CT826512010 with the Department of Environmental and Molecular Toxicology, North Caroline State University, Raleigh.The research described in this article has been reviewed by the U.S. Environmental Protection Agency, and approved for publication. Approval does not signify that the contents necessarily reflect the views and the policies of the Agency, nor does mention of trade names or commercial products constitute endorsement or recommendation for use.     

Seed Dispersal by a Frugivore Pycnonotus sinensis and the Distribution of Sambucus chinensis

Frugivorous birds play an important role in the distribution of those plants on whose fruits they feed and affect species composition of plant communities. The fruiting phenology of Sambucus chinensis Lindl. in Wuhan area ranges from July to November, while abundant fruits are available in July and August. Although other seven species of frugivores were found in the study area, only Pycnonotus sinensis (Gmelin), a resident bird endemic to China, was observed foraging in the fruits of this plant for the field. Pycnonotus sinensis shifts from eating fruits of Broussonetia papyrifera (L.) Vent. to those of S. chinensis from July to August. Identification of seeds in a total of 658 feces disseminated by P. sinensis which were collected in the field showed that most of the seeds came from S. chinensis and B. papyrifera. Seeds in those feces collected at the end of August were all of S. chinensis. Field investigation of the associations of S. chinensis shows that its frequent companion species are B. papyrifera, Cinnamomum camphora, Ligustrum lucidum and Rubus biflorus, whose fruits are also preferred by this frugivore in different seasons. The distribution of Pycnonotus sinensis is limited to the south of Qinling Mountain in China. The distribution of S. chinensis is basically consistent with that of Pycnonotus sinensis. The result indicates that seed dispersal of S. chinensis depends on the foraging of this frugivore. Likewise, the fruits of S. chinensis are the primary food for this frugivore in autumn. As the result suggests, conservationists and restoration biologists should highlight the study of plant-disperser mutualism in the future work.