Analysis of tryptamines in whole blood by UHPLC-MS/MS

Tryptaminer er en undergruppe av nye psykoaktive substanser (NPS) med økende forekomst i senere år. I tillegg kan en fornyet interesse for medisinsk bruk av tryptaminer føre til økt bruk i samfunnet. Dette gjør det aktuelt med metoder for analyse av tryptaminer. En Ultra-High Performance Liquid Chromatography Tandem Mass Spectrometry (UHPLCMS/MS)-metode ble utviklet for separasjon og deteksjon av 21 tryptaminer. Kromatografisk separasjon av analyttene ble utført ved omvendtfasekromatografi i bifenyl-kolonne (2,1 mm x 100 mm, 1,8 µm partikler) med kolonnetemperatur på 60 °C, mobilfase (MF)-hastighet på 0,5 mL/min, injiseringsvolum 1,5 µL og gradient eluering på 7,5 min med 10 mM ammounimformatbuffer med pH 3,1 og metanol som organisk løsemiddel. Deteksjon ble utført med elektrosprayionisering (ESI) i positiv modus og med multiple reaction monitoring (MRM)– skannmodus. Metoden ble brukt til å studere stabilitet av analyttene i fullblod og i stamløsninger i organiske løsningsmidler. For prøveopparbeidelse av tryptaminer i fullblod ble proteinutfelling (PPT) med acetonitril: metanol (85:15, v/v) benyttet. Stabilitetsprøver av tryptaminer i fullblod/organiske løsningsmidler oppbevart i romtemperatur, kjøleskap og fryser ble analysert med UHPLC-MS/MS. Gradienten førte til separasjon av 13 tryptaminer, mens parene 5-MeO-DPT og 5-MeO-DiPT, EPT og 5-MeO-EPT, MiPT og 4-AcO-DET, og 4-OH-DET og 4-OH-MiPT ikke ble kromatografisk separert. Metoden kunne ikke identifisere 4-OH-DET, 5-MeO-DiPT og 5- MeO-DPT. Stabilitetsstudien for 21 tryptaminer i renstoffløsninger viste at ved tilfeller hvor bruksløsningene blir stående i romtemperatur vil de for dette utvalget av tryptaminer allikevel kunne brukes senere, med unntak av de acetoksy (AcO)-substituerte tryptaminene. Stabilitetsstudien for 13 tryptaminer i fullblod viste at tiden mellom prøvetaking og rettstoksikologisk analyse ikke er kritisk for dette utvalget av tryptaminer, med unntak av de AcO-substituerte tryptaminene som bør analyseres uten mer ventetid enn nødvendig.Tryptamines are a category of New psychoactive substances (NPS) and its occurrence is increasing. In addition, a renewed interest in the medical use of tryptamines may cause increased use in society. Therefore, it is relevant with methods for analysis of tryptamines. An Ultra-High Performance Liquid Chromatography Tandem Mass Spectrometry (UHPLC-MS/MS) method was developed for the separation and detection of 21 tryptamines. The chromatographic separation of the analytes was performed by reverse phase chromatography in biphenyl column (2,1 mm x 100 mm, 1,8 μm particles) with a temperature of 60 °C, mobile phase flow 0,5 mL/min, injection volume 1,5 μL and gradient elution of 7,5 min with 10 mM ammounim format buffer with pH 3,1 and with methanol as organic modifier. Detection was performed with electron spray ionization (ESI) in positive mode and with multiple reaction monitoring (MRM) scan mode. The method was used for stability studies of the analytes in whole blood and in stock solutions in organic solvents. For sample preparation of tryptamines in whole blood protein precipitation (PPT) with acetonitrile: methanol (85:15, v/v) was used. Stability samples of tryptamines in whole blood/ organic solvents stored in room temperature, refrigerator and freezer was analyzed with UHPLC-MS/MS. The gradient separated 13 tryptamines, while 5-MeO-DPT and 5-MeO-DiPT, EPT and 5-MeO-EPT, MiPT and 4-AcO-DET, and 4-OH-DET and 4-OH-MiPT was not chromatographic separated. The method was not able to identify 4-OH-DET, 5-MeO-DiPT and 5-MeO-DPT. The stability study for 21 tryptamines in organic solvents, showed that in causes when the stock solutions are kept in room temperature they will still be able to be used again later for this selection of tryptamines, with the exception of the acetoxy substituted tryptamines. The stability study for 13 tryptamines in whole blood showed that the time between sampling and forensic toxicologic analysis is not critical for this selection of tryptamines, with the exception of the acetoxy substituted tryptamines that should be analyzed with no more waiting time than necessary.M-K

Clinical mastitis in ewes; bacteriology, epidemiology and clinical features

<p>Abstract</p> <p>Background</p> <p>Clinical mastitis is an important disease in sheep. The objective of this work was to identify causal bacteria and study certain epidemiological and clinical features of clinical mastitis in ewes kept for meat and wool production.</p> <p>Methods</p> <p>The study included 509 ewes with clinical mastitis from 353 flocks located in 14 of the 19 counties in Norway. Clinical examination and collection of udder secretions were carried out by veterinarians. Pulsed-field gel electrophoresis (PFGE) was performed on 92 <it>Staphylococcus aureus </it>isolates from 64 ewes.</p> <p>Results and conclusion</p> <p><it>S. aureus </it>was recovered from 65.3% of 547 clinically affected mammary glands, coagulase-negative staphylococci from 2.9%, enterobacteria, mainly <it>Escherichia coli</it>, from 7.3%, <it>Streptococcus </it>spp. from 4.6%, <it>Mannheimia haemolytica </it>from 1.8% and various other bacteria from 4.9%, while no bacteria were cultured from 13.2% of the samples. Forty percent of the ewes with unilateral clinical <it>S. aureus </it>mastitis also had a subclinical <it>S. aureus </it>infection in the other mammary gland. Twenty-four of 28 (86%) pairs of <it>S. aureus </it>isolates obtained from clinically and subclinically affected mammary glands of the same ewe were indistinguishable by PFGE. The number of identical pairs was significantly greater than expected, based on the distribution of different <it>S. aureus </it>types within the flocks. One-third of the cases occurred during the first week after lambing, while a second peak was observed in the third week of lactation. Gangrene was present in 8.8% of the clinically affected glands; <it>S. aureus </it>was recovered from 72.9%, <it>Clostridium perfringens </it>from 6.3% and <it>E. coli </it>from 6.3% of the secretions from such glands. This study shows that <it>S. aureus </it>predominates as a cause of clinical ovine mastitis in Norway, also in very severe cases. Results also indicate that <it>S. aureus </it>is frequently spread between udder halves of infected ewes.</p

Large-scale Molecular Diet Analysis in a Generalist Marine Mammal Reveals Male Preference for Prey of Conservation Concern

Sex-specific diet information is important in the determination of predator impacts on prey populations. Unfortunately, the diet of males and females can be difficult to describe, particularly when they are marine predators. We combined two molecular techniques to describe haul-out use and prey preferences of male and female harbor seals (Phoca vitulina) from Comox and Cowichan Bay (Canada) during 2012-2013. DNA metabarcoding quantified the diet proportions comprised of prey species in harbor seal scat, and qPCR determined the sex of the individual that deposited each scat. Using 287 female and 260 male samples, we compared the monthly sex ratio with GLMs and analyzed prey consumption relative to sex, season, site, and year with PERMANOVA. The sex ratio between monthly samples differed widely in both years (range = 12%-79% males) and showed different patterns at each haul-out site. Male and female diet differed across both years and sites: Females consumed a high proportion of demersal fish species while males consumed more salmonid species. Diet composition was related to both sex and season (PERMANOVA: R 2 = 27%, p \u3c 0.001; R 2 = 24%, p \u3c 0.001, respectively) and their interaction (PERMANOVA: R 2 = 11%, p \u3c 0.001). Diet differences between males and females were consistent across site and year, suggesting fundamental foraging differences, including that males may have a larger impact on salmonids than females. Our novel combination of techniques allowed for both prey taxonomic and spatiotemporal resolution unprecedented in marine predators

DNA-Based Diet Analysis for Any Predator

Background: Prey DNA from diet samples can be used as a dietary marker; yet current methods for prey detection require a priori diet knowledge and/or are designed ad hoc, limiting their scope. I present a general approach to detect diverse prey in the feces or gut contents of predators. Methodology/Principal Findings: In the example outlined, I take advantage of the restriction site for the endonuclease Pac I which is present in 16S mtDNA of most Odontoceti mammals, but absent from most other relevant non-mammalian chordates and invertebrates. Thus in DNA extracted from feces of these mammalian predators Pac I will cleave and exclude predator DNA from a small region targeted by novel universal primers, while most prey DNA remain intact allowing prey selective PCR. The method was optimized using scat samples from captive bottlenose dolphins (Tursiops truncatus) fed a diet of 6–10 prey species from three phlya. Up to five prey from two phyla were detected in a single scat and all but one minor prey item (2% of the overall diet) were detected across all samples. The same method was applied to scat samples from free-ranging bottlenose dolphins; up to seven prey taxa were detected in a single scat and 13 prey taxa from eight teleost families were identified in total. Conclusions/Significance: Data and further examples are provided to facilitate rapid transfer of this approach to any predator. This methodology should prove useful to zoologists using DNA-based diet techniques in a wide variety of study systems

Undersøkelse av utfordringer knyttet til bruk av flomsedimenter som prøvemedium i geokjemisk kartlegging - En studie av flomsedimenter i Gudbrandsdalen

Denne oppgaven tar for seg problemstillinger knyttet til bruk av flomsedimenter som prøvemedium, med utgangspunkt i Gudbrandsdalen som lokalitet. Flomsedimenter er antatt å reflektere geokjemien i et område da disse er dannet ved prosesser som forvitring, erosjon, transport og avsetting av bergarter og mineraler i nedslagsfeltet. Det eksisterer likevel en pågående diskusjon om flomsedimentene er representative da sammensetningen kan påvirkes av ulike fysiske og kjemiske prosesser. Oppgaven er skrevet i samarbeid med Norges geologiske undersøkelse. Det ble samlet inn prøver ved totalt ti prøvepunkter av flomsedimenter ved ulike dybder i forskjellige profiler. Dypeste prøve ble tatt ved omtrent 80 cm dybde. Det ble også samlet inn prøver av elvesedimenter og filterprøver av partikulært elvemateriale, hvor dette lot seg gjøre. Prøvene ble tørket ved romtemperatur, i tørkeskap eller ved frysetørking avhengig av fuktinnhold. Sedimentene ble dekomponert med 9 mL 50 % v/v HNO3 i UltraCLAVE, mens filterprøvene ble dekomponert med 3 mL 50 % v/v HNO3. Alle prøvene ble analysert med HR-ICP-MS. Det ble også bestemt glødetap på et utvalg topprøver langs Gudbrandsdalslågen, samt nedover i én profil. Sammensetningen til flomsedimentene og tilhørende elvesedimenter ble sammenlignet og det ble funnet en relativt lik sammensetning av hovedelementene; Fe, Al, Mg, Ca, Si, Na, K og Mn. På bakgrunn av disse resultatene kan det konkluderes med at flomsedimenter og elvesedimenter er et resultat av de samme sedimentkildene, og at flomsedimenter dermed er like representative for geokjemien som elvesedimenter. Flomsedimentprofilene viste litt ulike trender mellom de forskjellige prøvepunktene, noe som indikerer bidrag fra ulike aktive sedimentkilder og forurensningskilder langs elva. Lagene nedover i en profil viste en forholdvis lik sammensetning mellom hovedelementene. Dette er mest sannsynlig et resultat av små endringer i bidrag fra sedimentkilder ved ulike flomepisoder. Det ble ved flere prøvepunkt observert en nedgang i konsentrasjon av ulike sporelementer som Zn, Cu, Cr, Ni, Cd, Pb og As oppover i profilene. Dette var noe uventet da en økning i konsentrasjon ville vært forventet som følge av antropogene bidrag. Ved spesielt ett prøvepunkt ble det funnet antydninger til vertikal migrasjon av elementene Cd og Ni. Basert på dette kan det stilles spørsmål ved om de dyptliggende sedimentlagene kan benyttes til å bestemme naturlig bakgrunnsnivå ved det gitte prøvepunktet. Resultatene antyder også at valg av prøvepunkt vil være avgjørende for oppnådd resultat

Analyse av tryptaminer i fullblod med UHPLC-MS/MS

Tryptaminer er en undergruppe av nye psykoaktive substanser (NPS) med økende forekomst i senere år. I tillegg kan en fornyet interesse for medisinsk bruk av tryptaminer føre til økt bruk i samfunnet. Dette gjør det aktuelt med metoder for analyse av tryptaminer. En Ultra-High Performance Liquid Chromatography Tandem Mass Spectrometry (UHPLCMS/MS)-metode ble utviklet for separasjon og deteksjon av 21 tryptaminer. Kromatografisk separasjon av analyttene ble utført ved omvendtfasekromatografi i bifenyl-kolonne (2,1 mm x 100 mm, 1,8 µm partikler) med kolonnetemperatur på 60 °C, mobilfase (MF)-hastighet på 0,5 mL/min, injiseringsvolum 1,5 µL og gradient eluering på 7,5 min med 10 mM ammounimformatbuffer med pH 3,1 og metanol som organisk løsemiddel. Deteksjon ble utført med elektrosprayionisering (ESI) i positiv modus og med multiple reaction monitoring (MRM)– skannmodus. Metoden ble brukt til å studere stabilitet av analyttene i fullblod og i stamløsninger i organiske løsningsmidler. For prøveopparbeidelse av tryptaminer i fullblod ble proteinutfelling (PPT) med acetonitril: metanol (85:15, v/v) benyttet. Stabilitetsprøver av tryptaminer i fullblod/organiske løsningsmidler oppbevart i romtemperatur, kjøleskap og fryser ble analysert med UHPLC-MS/MS. Gradienten førte til separasjon av 13 tryptaminer, mens parene 5-MeO-DPT og 5-MeO-DiPT, EPT og 5-MeO-EPT, MiPT og 4-AcO-DET, og 4-OH-DET og 4-OH-MiPT ikke ble kromatografisk separert. Metoden kunne ikke identifisere 4-OH-DET, 5-MeO-DiPT og 5- MeO-DPT. Stabilitetsstudien for 21 tryptaminer i renstoffløsninger viste at ved tilfeller hvor bruksløsningene blir stående i romtemperatur vil de for dette utvalget av tryptaminer allikevel kunne brukes senere, med unntak av de acetoksy (AcO)-substituerte tryptaminene. Stabilitetsstudien for 13 tryptaminer i fullblod viste at tiden mellom prøvetaking og rettstoksikologisk analyse ikke er kritisk for dette utvalget av tryptaminer, med unntak av de AcO-substituerte tryptaminene som bør analyseres uten mer ventetid enn nødvendig

Livssyklusanalyser og bærekraftige omstillinger

The social and political barriers to sustainability are often presumed to boil down to misalignments of power and motivation. If only those who have the power to initiate sustainability transitions could be motivated to do so – or conversely; those who have the motivation could be empowered to do so – then sustainability would presumably follow. This presumption is problematic. Not only because it skirts the difficulties in operationalizing and evaluating sustainability. But also because it neglects the social and political processes through which some behaviors and technologies come to be coded as ‘sustainable’ and others as ‘unsustainable’. Over the past few decades, life cycle assessments (LCA) have come to take center-stage in these processes. The upside of this trend has been a shift in emphasis, away from face-sustain-ability1 and direct emissions; towards supply chains and displacement effects. The downsides have included all the predictable pitfalls of governance by numbers: operational definitions with poor construct validity; black boxes that are hard for non-experts to challenge; quarrels over conflicting evidence, etc. Over the same period, an extensive literature has accrued, dealing with the validity, limitations, and appropriate use of LCA. The impacts particular LCAs have had on particular sustainability discourses and decision-making processes, by contrast, have largely been neglected as a topic of research. This thesis seeks to bridge this hole in the extant literature, by examining the impacts LCA has had on Norwegian sustainability discourse and policy-making; by synthesizing insights from social constructivism (i.e. how taken-for-granted realities congeal through social practices), science and technology studies (e.g. black-boxing, epistemic authority, and boundary work), and the ethics and sociology of quantifications (e.g. mathiness, metric fixation, and data inertia). The thesis begins by assessing the validity of LCA through a critical appraisal of the extant literature on climate metrics and life cycle assessments. It then goes on to describe the role of LCA in sustainability discourses through two case studies: the 2021 debate about a managed decline of petroleum production; and the 2007 - 2019 debate about a managed transition from plastic to paper bags. Finally, it addresses the role of LCA in policymaking through a combination of document analysis and unstructured interviews with representatives of local authorities in the Norwegian county of Rogaland. This examination provides a basis for arguing that: (1) the boundaries between ‘sustainable’ and ‘unsustainable’ activities cannot in general be drawn objectively – but they nonetheless have to be drawn somehow; (2) LCA can serve this purpose and may facilitate rational decision making; (3) but tends – as a general rule – to be short on validity and susceptible to reification, co-option, and technocratic decision making

Livssyklusanalyser og bærekraftige omstillinger

The social and political barriers to sustainability are often presumed to boil down to misalignments of power and motivation. If only those who have the power to initiate sustainability transitions could be motivated to do so – or conversely; those who have the motivation could be empowered to do so – then sustainability would presumably follow. This presumption is problematic. Not only because it skirts the difficulties in operationalizing and evaluating sustainability. But also because it neglects the social and political processes through which some behaviors and technologies come to be coded as ‘sustainable’ and others as ‘unsustainable’. Over the past few decades, life cycle assessments (LCA) have come to take center-stage in these processes. The upside of this trend has been a shift in emphasis, away from face-sustain-ability1 and direct emissions; towards supply chains and displacement effects. The downsides have included all the predictable pitfalls of governance by numbers: operational definitions with poor construct validity; black boxes that are hard for non-experts to challenge; quarrels over conflicting evidence, etc. Over the same period, an extensive literature has accrued, dealing with the validity, limitations, and appropriate use of LCA. The impacts particular LCAs have had on particular sustainability discourses and decision-making processes, by contrast, have largely been neglected as a topic of research. This thesis seeks to bridge this hole in the extant literature, by examining the impacts LCA has had on Norwegian sustainability discourse and policy-making; by synthesizing insights from social constructivism (i.e. how taken-for-granted realities congeal through social practices), science and technology studies (e.g. black-boxing, epistemic authority, and boundary work), and the ethics and sociology of quantifications (e.g. mathiness, metric fixation, and data inertia). The thesis begins by assessing the validity of LCA through a critical appraisal of the extant literature on climate metrics and life cycle assessments. It then goes on to describe the role of LCA in sustainability discourses through two case studies: the 2021 debate about a managed decline of petroleum production; and the 2007 - 2019 debate about a managed transition from plastic to paper bags. Finally, it addresses the role of LCA in policymaking through a combination of document analysis and unstructured interviews with representatives of local authorities in the Norwegian county of Rogaland. This examination provides a basis for arguing that: (1) the boundaries between ‘sustainable’ and ‘unsustainable’ activities cannot in general be drawn objectively – but they nonetheless have to be drawn somehow; (2) LCA can serve this purpose and may facilitate rational decision making; (3) but tends – as a general rule – to be short on validity and susceptible to reification, co-option, and technocratic decision making

Opprettholdelse av aktivitetsidentitet på lang sikt etter hjerneslag: en systematisk litteraturgjennomgang

Bakgrunn: 12 000 rammes årlig av hjerneslag i Norge og det kan føre til sensoriske, motoriske og kognitive utfall. Opplevde funksjonsendringer kan påvirke deltakelse i aktiviteter, som vil virke inn på hvordan man ser på seg selv som aktivt menneske. Aktivitetsidentitet er et ergoterapeutisk perspektiv som tar for seg hvordan mennesker danner et bilde av hvem man er og ønsker å være, basert på aktivitetene de deltar i. Hensikt: Formålet med denne bacheloroppgaven er å se på hvordan voksne etter hjerneslag kan opprettholde sin aktivitetsidentitet på lang sikt. Dette skal diskuteres opp mot ergoterapeutisk teori, vitenskapelig forskning og annen kunnskap. Metode: Med utgangspunkt i scoping review ble det gjort et systematisk litteratursøk. Der ble det innhentet kvalitativ forskning fra Medline, PubMed og AMED. Det ble inkludert fem forskningsartikler, hvor dataen ble analysert og relevante funn ble trukket frem. Resultat: Gjennomgående i artiklene kommer det frem hvordan identiteten og selvopplevelsen til voksne blir påvirket etter et hjerneslag. Flere av artiklene tar for seg prosessen etter et hjerneslag som kompleks og varierende for den enkelte. Strategier og støtte fra andre rundt seg, for å håndtere den nye livssituasjonen, blir også trukket frem. Konklusjon: For voksne etter hjerneslag er endringene i livssituasjonen omfattende, noe som påvirker selvtillit, selvopplevelse, mestring og motivasjon. Ergoterapeuter kan bidra med å støtte voksne i å finne nye aktiviteter og å utvikle individuelle rehabiliteringsplaner, som kan bidra til å støtte individer i å bygge en ny hverdag. Kontinuerlig deltakelse i meningsfulle aktiviteter vil være nødvendig for å kunne opprettholde aktivitetsidentiteten etter hjerneslag for voksne på lang sikt. Nøkkelord: hjerneslag, aktivitetsidentitet, voksne, hverdag, senfase

In what ways are designers' and operators' reasonable-world assumptions not reasonable assumptions?

Although the technical reliability of complex process plant generally continues to improve, such systems remain vulnerable to unmet assumptions on the part of designers, operators and maintenance staff. The purpose of this study was to characterize the nature of these assumptions, particularly those that were implied in people's activity rather than those that were explicitly invoked. Inferences about the assumptions were made from a set of accident reports. This characterization was then used as the basis of a tool that could help people examine how either their own or other peoples’ assumptions could imperil a system. Such a tool both structures the process of testing assumptions and helps synthesize the knowledge provided by historical failures. The tool was evaluated with a variety of organizations and used in a case study, where it contributed to the process of risk analysis in a variety of ways