48 research outputs found

    Metabolic profiling of zebrafish embryo development from blastula period to early larval stages

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    The zebrafish embryo is a popular model for drug screening, disease modelling and molecular genetics. In this study, samples were obtained from zebrafish at different developmental stages. The stages that were chosen were 3/4, 4/5, 24, 48, 72 and 96 hours post fertilization (hpf). Each sample included fifty embryos. The samples were analysed using gas chromatography time-of-flight mass spectrometry (GC-TOF-MS). Principle component analysis (PCA) was applied to get an overview of the data and orthogonal projection to latent structure discriminant analysis (OPLS-DA) was utilised to discriminate between the developmental stages. In this way, changes in metabolite profiles during vertebrate development could be identified. Using a GC-TOF-MS metabolomics approach it was found that nucleotides and metabolic fuel (glucose) were elevated at early stages of embryogenesis, whereas at later stages amino acids and intermediates in the Krebs cycle were abundant. This agrees with zebrafish developmental biology, as organs such as the liver and pancreas develop at later stages. Thus, metabolomics of zebrafish embryos offers a unique opportunity to investigate large scale changes in metabolic processes during important developmental stages in vertebrate development. In terms of stability of the metabolic profile and viability of the embryos, it was concluded at 72 hpf was a suitable time point for the use of zebrafish as a model system in numerous scientific applications

    Boldness Predicts Social Status in Zebrafish (Danio rerio)

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    This study explored if boldness could be used to predict social status. First, boldness was assessed by monitoring individual zebrafish behaviour in (1) an unfamiliar barren environment with no shelter (open field), (2) the same environment when a roof was introduced as a shelter, and (3) when the roof was removed and an unfamiliar object (Lego® brick) was introduced. Next, after a resting period of minimum one week, social status of the fish was determined in a dyadic contest and dominant/subordinate individuals were determined as the winner/loser of two consecutive contests. Multivariate data analyses showed that males were bolder than females and that the behaviours expressed by the fish during the boldness tests could be used to predict which fish would later become dominant and subordinate in the ensuing dyadic contest. We conclude that bold behaviour is positively correlated to dominance in zebrafish and that boldness is not solely a consequence of social dominance

    Hexabromocyclododecanes (HBCDs) in the environment and humans: A review

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    Hexabromocyclododecanes (HBCDs) are brominated aliphatic cyclic hydrocarbons used as flame retardants in thermal insulation building materials, upholstery textiles, and electronics. As a result of their widespread use and their physical and chemical properties, HBCDs are now ubiquitous contaminants in the environment and humans. This review summarizes HBCD concentrations in several environmental compartments and analyzes these data in terms of point sources versus diffuse sources, biomagnification potential, stereoisomer profiles, time trends, and global distribution. Generally, higher concentrations were measured in samples (air, sediment, and fish) collected near point sources (plants producing or processing HBCDs), while lower concentrations were recorded in samples from locations with no obvious sources of HBCDs. High concentrations were measured in top predators, such as marine mammals and birds of prey (up to 9600 and 19 200 ng/g lipid weight, respectively), suggesting a biomagnification potential for HBCDs. Relatively low HBCD concentrations were reported in the few human studies conducted to date (median values varied between 0.35 and 1.1 ng/g lipid weight). HBCD levels in biota are increasing slowly and seem to reflect the local market demand. One important observation is the shift from the high percentage of the gamma-HBCD stereoisomer in the technical products to a dominance of the alpha-HBCD stereoisomer in biological samples. A combination of factors such as variations in solubility, partitioning behavior, uptake, and, possibly, selective metabolism of individual isomers may explain the observed changes in stereoisomer patterns. Recommendations for further work include research on how HBCDs are transferred from products into the environment upon production, use, and disposal. Time trends need to be analyzed more in detail, including HBCD stereoisomers, and more data on terrestrial organisms are needed, especially for humans. Whenever possible, HBCDs should be analyzed as individual stereoisomers in order to address their fate and effects

    The 42nd Symposium Chromatographic Methods of Investigating Organic Compounds : Book of abstracts

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    The 42nd Symposium Chromatographic Methods of Investigating Organic Compounds : Book of abstracts. June 4-7, 2019, Szczyrk, Polan

    QSBMR Quantitative Structure Biomagnification Relationships : Studies Regarding Persistent Environmental Pollutants in the Baltic Sea Biota

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    I have studied persistent environmental pollutants in herring (Clupea harengus), in adult guillemot (Uria aalge) and in guillemot eggs from the Baltic Sea. The studied contaminants were organochlorines (OCs); dichlorodiphenyltrichloroethanes (DDTs), polychlorinated biphenyls (PCBs), hexachlorobenzene (HCB), hexachlorocyclohexanes (HCHs), and brominated flame retardants (BFRs); polybrominated diphenylethers (PBDEs) and hexabromocyclododecane (HBCD). The highest concentration in both species was shown by p,p′DDE with a concentration in guillemot egg (geometric mean (GM) with 95% confidence interval) of 18200 (17000 – 19600) ng/g lipid weight. The BFR with the highest concentration in guillemot egg was HBCD with a GM concentration of 140 (120 – 160) ng/g lw. To extract additional and essential information from the data, not possible to obtain using only univariate or bivariate statistics, I used multivariate data analysis techniques; principal components analysis (PCA), partial least squares regression (PLS), soft independent modelling of class analogy (SIMCA), and PLS discriminant analysis (PLS-DA). I found e.g.; that there are significant negative correlations between egg weight and the concentrations of HCB and p,p'DDE; that concentrations of OCs and BFRs in the organisms co-varied so that concentrations of OCs can be used to calculate concentrations of BFRs; and, that several contaminants (e.g. HBCD) had higher concentration in guillemot egg than in guillemot muscle, that several (e.g. BDE47) showed no concentration difference between muscle and egg and that one contaminant (BDE154) showed higher concentration in the guillemot muscles than in egg. In this thesis I developed a new method, “randomly sampled ratios” (RSR), to calculate biomagnification factors (BMFs) i.e. the ratio between the concentration of a contaminant in an organism and the concentration of the same contaminant in its food. With this new method BMFs are denoted with an estimate of variation. Contaminants that biomagnify are e.g., p,p′DDE, CB118, HCB, βHCH and all of the BFRs. Those that do not biomagnify are e.g., p,p′DDT, αHCH and CB101. Lastly, to investigate which of the contaminants descriptors (physical-chemical/other properties and characteristics) that correlates to the biomagnification of the contaminants, I modeled the contaminants’ respective BMFRSR versus ~100 descriptors and showed that ~20 descriptors in combination were important, either favoring or counteracting biomagnification between herring and guillemot

    Significant changes in metabolic profiles after intervention with selenium and coenzyme q10 in an elderly population

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    Selenium and coenzyme Q10 (SeQ10) are important for normal cellular function. Low selenium intake leads to increased cardiovascular mortality. Intervention with these substances with healthy elderly persons over a period of four years in a double-blind, randomised placebo-controlled prospective study showed reduced cardiovascular mortality, increased cardiac function, and a lower level of NT-proBNP. Therefore, we wanted to evaluate changes in biochemical pathways as a result of the intervention with SeQ10 using metabolic profiling. From a population of 443 healthy elderly individuals that were given 200 µg selenium and 200 mg coenzyme Q10, or placebo daily for four years, we selected nine males on active intervention and nine males on placebo for metabolic profiling in the main study. To confirm the results, two validation studies (study 1 n = 60 males, study 2 n = 37 males) were conducted. Principal component analyses were used on clinical and demographic data to select representative sets of samples for analysis and to divide the samples into batches for analysis. Gas chromatography time-of-flight mass spectrometry-based metabolomics was applied. The metabolite data were evaluated using univariate and multivariate approaches, mainly T-tests and orthogonal projections to latent structures (OPLS) analyses. Out of 95 identified metabolites, 19 were significantly decreased due to the intervention after 18 months of intervention. Significant changes could be seen in the pentose phosphate, the mevalonate, the beta-oxidation and the xanthine oxidase pathways. The intervention also resulted in changes in the urea cycle, and increases in the levels of the precursors to neurotransmitters of the brain. This adds information to previous published results reporting decreased oxidative stress and inflammation. This is the first-time metabolic profiling has been applied to elucidate the mechanisms behind an intervention with SeQ10. The study is small and should be regarded as hypothesis-generating; however, the results are interesting and, therefore, further research in the area is needed. This study was registered at Clinicaltrials.gov, with the identifier NCT01443780.The analysis costs were partially supported by grants from Pharma Nord Aps, Denmark, the County Council of Östergötland, Linköping University. The funding organisations had no role in the design, management, analysis, or interpretation of the data, nor in the preparation, review or approval of the manuscript. No economic compensation was distributed.publishedVersio
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