DNA damage and health effects in juvenile haddock exposed to sediment or produced water associated PAHs

Tilstandsundersøkelsene i Nordsjøen har det siste tiår vist gentoksiske effekter (DNA-addukter) i fisk samlet inn i områder med offshore olje- og gassvirksomhet. Kilden og identiteten til de gentoksiske forbindelsene har til nå ikke blitt identifisert. Målsetningen for prosjektet har vært å studere dannelsen av DNA-skade i hyse som utsettes for ulike petrogene eller pyrogene polyaromatiske hydrokarboner (PAH). Målet har vært å identifisere kildene til forurensningen som medfører DNA-addukter observert i hyse fanget rundt oljefeltene i Nordsjøen. Dannelsen av DNA-skade over tid i hyse under kronisk eksponering for PAH og andre oljehydrokarboner fra følgende kilder er blitt studert: Ekstrakter av produsert vann (Statfjord A); destillasjonsfraksjoner av råolje fra Gullfaks (representere oljebasert borevæske); pyrogene PAH. Denne rapporten presenterer resultatene av studiene på DNA addukter i hyse og et utvalg av andre biologiske effektparametre.The Condition Monitoring in the North Sea the last ten years have documented genotoxic effects (DNA adduct) in fish collected in areas with extensive offshore oil and gas activity. However, the source and identity of genotoxic compound has not been identified. The objective of this project has been to study the formation of DNA damage in haddock exposed to petrogenic or pyrogenic polyaromatic hydrocarbons (PAHs) from different sources: Extracts of produced water (Statfjord A); distillation fractions of crude oil from Gullfaks (representing oil based drilling mud); pyrogenic PAH This report presents all the results from the study on DNA adducts and a selection of other biological effect parameters

Etablering av metode for anrikning og deteksjon av sirkulerende tumorceller i perifert blod

Master's thesis in Biological chemistryKreftceller som sirkulerer med perifert blod kalles sirkulerende tumorceller (STC). Under de rette forholdene kan disse gi opphav til nye svulster i andre organer (spredning). Kreft i bukspyttkjertelen er den fjerde største kreftrelaterte dødsårsaken i Norge, noe som i hovedsak skyldes at den er vanskelig å oppdage samt ineffektiv kjemoterapibehandling. Forskning tyder på at påvisning av STC kan gi klinisk relevant informasjon ved en rekke kreftformer. Hensikten med denne masteroppgaven var derfor å etablere en metode for anrikning og deteksjon av STC fra perifert blod, da tilknyttet bukspyttkjertelkreft. Oppgaven inngår i et større forskningsprosjekt ved Stavanger Universitetssykehus i regi av forskningsgruppen for kreft og medisinsk fysikk ved Avdeling for blod og kreftsykdommer. Blodprøven tas på EDTA-rør og anrikningsmetoden er bruk av tetthetsgradientmediet LymphoprepTM. Disse ble kvalitetsvurdert og det ble funnet lite tap av mononukleære celler og celleaggregat. Holdbarheten til EDTA-blod ble fastsatt til maksimalt 48 timers henstand før anrikning av STC bør utføres. Deteksjonsmetoden er RT rt-qPCR hvor epitelspesifikke mRNA benyttes som surrogatbiomarkører for STC. De potensielle biomarkørene som ble etablert og undersøkt var CEACAM5, CEACAM6, CK8, CK19, EPCAM, MMP11 og PRSS1. Basert på undersøkelser av tre cellelinjer fra svulster i bukspyttkjertel og normalt blod, viste biomarkørene CEACAM5, CK8, CK19 og EPCAM beste potensial for STC deteksjon

Identification of an albumin-like protein in plasma of Atlantic cod (Gadus morhua) and its biomarker potential for PAH contamination

Increased research efforts are currently focusing on Atlantic cod (Gadus morhua) and its significance for monitoring the contaminant situation in marine environments. Polycyclic aromatic hydrocarbons (PAHs) are well known toxic and carcinogenic compounds, thus continuous monitoring is required to ensure ecosystem sustainability and human food safety. A sensitive biomarker of PAH exposure in humans is the detection of PAH metabolites bound to albumin in blood. The potential of a similar PAH-albumin biomarker in Atlantic cod was therefore investigated by a desktop bioinformatic study followed by liquid chromatography mass spectrometry/mass spectrometry analysis of plasma from 16 fish. For the first time, an albumin-like protein in plasma of Atlantic cod is described, and the biomarker potential based on PAH-albumin adduct detection is discussed. Due to the detected low abundance of the albumin-like protein, it was found unlikely to be applicable as a new biomarker tool for evaluation of PAH exposure

Biological effects of polycyclic aromatic hydrocarbons (PAH) and their first metabolic products in in vivo exposed Atlantic cod ( Gadus morhua )

International audienceThe monitoring of the presence of polycyclic aromatic hydrocarbons (PAH) in the aquatic environment is a worldwide activity since some of these compounds are well-established carcinogens and mutagens. Contaminants in this class are in fact regarded as priority hazardous substances for environmental pollution (Water Framework Directive 2000/60/EC). In this study, Atlantic cod (Gadus morhua) was selected to assess in vivo effects of two PAH and their first metabolic products, namely, the corresponding trans-dihydrodiols, using biological markers. Fish were exposed for 1 wk to a single PAH (naphthalene or chrysene) and its synthetic metabolites ((1R,2R)-1,2-dihydronaphthalene-1,2-diol and (1R,2R)-1,2-dihydrochrysene-1,2-diol) by intraperitoneal injection in a continuous seawater flow system. After exposure, PAH metabolism including PAH metabolites in bile and ethoxyresorufin O-deethylase (EROD) activity, oxidative stress glutathione S-transferases (GST) and catalase (CAT) activities, and genotoxicity such as DNA adducts were evaluated, as well as general health conditions including condition index (CI), hepatosomatic index (HSI), and gonadosomatic index (GSI). PAH metabolite values were low and not significantly different when measured with the fixed-wavelength fluorescence screening method, while the gas chromatography-mass spectroscopy (GC-MS) method showed an apparent dose response in fish exposed to naphthalene. DNA adduct levels ≥0.16 × 10(-8) relative adduct level (RAL) were detected. It should be noted that 0.16 × 10(-8) RAL is considered the maximal acceptable background level for this species. The other biomarkers activities of catalase, GST, and EROD did not display a particular compound- or dose-related response. The GSI values were significantly lower in some chrysene- and in both naphthalene- and naphthalene diol-exposed groups compared to control

Biological effects of polycyclic aromatic hydrocarbons (PAH) and their first metabolic products in in vivo exposed Atlantic cod ( Gadus morhua

International audienceThe monitoring of the presence of polycyclic aromatic hydrocarbons (PAH) in the aquatic environment is a worldwide activity since some of these compounds are well-established carcinogens and mutagens. Contaminants in this class are in fact regarded as priority hazardous substances for environmental pollution (Water Framework Directive 2000/60/EC). In this study, Atlantic cod (Gadus morhua) was selected to assess in vivo effects of two PAH and their first metabolic products, namely, the corresponding trans-dihydrodiols, using biological markers. Fish were exposed for 1 wk to a single PAH (naphthalene or chrysene) and its synthetic metabolites ((1R,2R)-1,2-dihydronaphthalene-1,2-diol and (1R,2R)-1,2-dihydrochrysene-1,2-diol) by intraperitoneal injection in a continuous seawater flow system. After exposure, PAH metabolism including PAH metabolites in bile and ethoxyresorufin O-deethylase (EROD) activity, oxidative stress glutathione S-transferases (GST) and catalase (CAT) activities, and genotoxicity such as DNA adducts were evaluated, as well as general health conditions including condition index (CI), hepatosomatic index (HSI), and gonadosomatic index (GSI). PAH metabolite values were low and not significantly different when measured with the fixed-wavelength fluorescence screening method, while the gas chromatography-mass spectroscopy (GC-MS) method showed an apparent dose response in fish exposed to naphthalene. DNA adduct levels ≥0.16 × 10(-8) relative adduct level (RAL) were detected. It should be noted that 0.16 × 10(-8) RAL is considered the maximal acceptable background level for this species. The other biomarkers activities of catalase, GST, and EROD did not display a particular compound- or dose-related response. The GSI values were significantly lower in some chrysene- and in both naphthalene- and naphthalene diol-exposed groups compared to control