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1.
  • Lind, Ulrika, et al. (författare)
  • Analysis of aquaporins from the euryhaline barnacle Balanus improvisus reveals differential expression in response to changes in salinity
  • 2017
  • Ingår i: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 12:7
  • Tidskriftsartikel (refereegranskat)abstract
    • Barnacles are sessile macro-invertebrates, found along rocky shores in coastal areas worldwide. The euryhaline bay barnacle Balanus improvisus (Darwin, 1854) (= Amphibalanus improvisus) can tolerate a wide range of salinities, but the molecular mechanisms underlying the osmoregulatory capacity of this truly brackish species are not well understood. Aquaporins are pore-forming integral membrane proteins that facilitate transport of water, small solutes and ions through cellular membranes, and that have been shown to be important for osmoregulation in many organisms. The knowledge of the function of aquaporins in crustaceans is, however, limited and nothing is known about them in barnacles. We here present the repertoire of aquaporins from a thecostracan crustacean, the barnacle B. improvisus, based on genome and transcriptome sequencing. Our analyses reveal that B. improvisus contains eight genes for aquaporins. Phylogenetic analysis showed that they represented members of the classical water aquaporins (Aqp1, Aqp2), the aquaglyceroporins (Glp1, Glp2), the unorthodox aquaporin (Aqp12) and the arthropod-specific big brain aquaporin (Bib). Interestingly, we also found two big brain-like proteins (BibL1 and BibL2) constituting a new group of aquaporins not yet described in arthropods. In addition, we found that the two water-specific aquaporins were expressed as C-terminal splice variants. Heterologous expression of some of the aquaporins followed by functional characterization showed that Aqp1 transported water and Glp2 water and glycerol, agreeing with the predictions of substrate specificity based on 3D modeling and phylogeny. To investigate a possible role for the B. improvisus aquaporins in osmoregulation, mRNA expression changes in adult barnacles were analysed after long-term acclimation to different salinities. The most pronounced expression difference was seen for AQP1 with a substantial (>100-fold) decrease in the mantle tissue in low salinity (3 PSU) compared to high salinity (33 PSU). Our study provides a base for future mechanistic studies on the role of aquaporins in osmoregulation. © 2017 Lind et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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2.
  • Larsson, Dennis, et al. (författare)
  • Antagonistic effects of 24R,25-dihydroxyvitamin D3 and 25-hydroxyvitamin D3 on L-type Ca2+ channels and Na+/Ca2+ exchange in enterocytes from Atlantic cod (Gadus morhua).
  • 2002
  • Ingår i: Journal of Molecular Endocrinology. - : Bioscientifica. - 0952-5041 .- 1479-6813. ; 28:1, s. 53-68
  • Tidskriftsartikel (refereegranskat)abstract
    • There is mounting evidence that vitamin D and its metabolites play important roles in regulating plasma calcium concentrations in teleost fish as in other vertebrates. The aims of the present study were to elucidate the possible cellular target mechanisms for the rapid actions of 24R,25(OH)(2)D(3), 25(OH)D(3) and 1,25(OH)(2)D(3) in Atlantic cod enterocytes at physiological doses, and to establish the concentration and thus the physiological range of circulating 24R,25(OH)(2)D(3), 25(OH)D(3) and 1,25(OH)(2)D(3) in the Atlantic cod. The plasma concentrations of 25(OH)D(3), 1,25(OH)(2)D(3) and 24R,25(OH)(2)D(3) were 15.3 +/- 2.7nM, 125.1 +/- 12.3pM and 10.1 +/- 23.5nM respectively. Exposure of enterocytes to 10mM calcium (Ca(2+)) evoked an increase in intracellular Ca(2+) concentrations ([Ca(2+)](i)). This increase was suppressed by 24R,25(OH)(2)D(3) dose-dependently, with an EC(50) of 4.9nM and a maximal inhibition of 60%. 24R,25(OH)(2)D(3) (20nM) abolished an increase in [Ca(2+)](i) (approximately 252%) in the control enterocytes exposed to 10microM S(-)-BAYK-8644, suggesting that the hormone acts by inhibiting Ca(2+) entry through L-type voltage-gated Ca(2+) channels. Administration of 20nM 24R,25(OH)(2)D(3) to enterocytes in the absence of extracellular Ca(2+) increased [Ca(2+)](i) by approximately 20%, indicating a release of Ca(2+) from intracellular stores. Administration of 25(OH)D(3) (20nM) resulted in a biphasic change in the enterocyte [Ca(2+)](i): within 1--5s, it decreased to 87 +/- 12nM below its mean basal [Ca(2+)](i) (334 +/- 13nM), followed by a rapid recovery of [Ca(2+)](i) to a new level, 10% lower than the initial [Ca(2+)](i). The rapid decrease, the recovery rate and the final [Ca(2+)](i) were all affected dose-dependently by 25(OH)D(3), with EC(50) values of 8.5, 17.0 and 18.9nM respectively. Furthermore, the effects of 25(OH)D(3) were sensitive to sodium (Na(+)), bepridil (10microM) and nifedipine (5 microM), suggesting that 25(OH)D(3) regulates the activity of both basolateral membrane-associated Na(+)/Ca(2+) exchangers and brush border membrane-associated L-type Ca(2+) channels. Administration of 25(OH)D(3) (10nM) to enterocytes in the absence of extracellular Ca(2+) increased [Ca(2+)](i) by approximately 18%, indicating a release of Ca(2+) from intracellular stores. 1,25(OH)(2)D(3) also affected enterocyte [Ca(2+)](i) in a biphasic manner: the rapid decrease, the recovery rate, and the mean final [Ca(2+)](i) were all affected dose-dependently, with EC(50) values of 8.3, 24.5 and 7.7nM respectively. The high EC(50) values for 1,25(OH)(2)D(3) compared with circulating concentrations of 1,25(OH)(2)D(3) (130pM) suggest that this effect is pharmacological, rather than of physiological relevance in enterocyte Ca(2+) homeostasis of the Atlantic cod. It is concluded that 24R,25(OH)(2)D(3) has a physiological role in decreasing intestinal Ca(2+) uptake via inactivation of L-type Ca(2+) channels, whereas the physiological role of 25(OH)D(3) is to increase enterocyte Ca(2+) transport via activation of Na(+)/Ca(2+) exchangers, concurrent with activation of L-type Ca(2+) channels.
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3.
  • Lind, Ulrika, et al. (författare)
  • Molecular Characterization of the alpha-Subunit of Na+/K+ ATPase from the Euryhaline Barnacle Balanus improvisus Reveals Multiple Genes and Differential Expression of Alternative Splice Variants
  • 2013
  • Ingår i: Plos One. - : Public Library of Science (PLoS). - 1932-6203. ; 8:10
  • Tidskriftsartikel (refereegranskat)abstract
    • The euryhaline bay barnacle Balanus improvisus has one of the broadest salinity tolerances of any barnacle species. It is able to complete its life cycle in salinities close to freshwater (3 PSU) up to fully marine conditions (35 PSU) and is regarded as one of few truly brackish-water species. Na+/K+ ATPase (NAK) has been shown to be important for osmoregulation when marine organisms are challenged by changing salinities, and we therefore cloned and examined the expression of different NAKs from B. improvisus. We found two main gene variants, NAK1 and NAK2, which were approximately 70% identical at the protein level. The NAK1 mRNA existed in a long and short variant with the encoded proteins differing only by 27 N-terminal amino acids. This N-terminal stretch was coded for by a separate exon, and the two variants of NAK1 mRNAs appeared to be created by alternative splicing. We furthermore showed that the two NAK1 isoforms were differentially expressed in different life stages and in various tissues of adult barnacle, i.e the long isoform was predominant in cyprids and in adult cirri. In barnacle cyprid larvae that were exposed to a combination of different salinities and pCO(2) levels, the expression of the long NAK1 mRNA increased relative to the short in low salinities. We suggest that the alternatively spliced long variant of the Nak1 protein might be of importance for osmoregulation in B. improvisus in low salinity conditions.
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4.
  • Rosengren, Malin, 1982, et al. (författare)
  • Born to be wild: effects of rearing density and environmental enrichment on stress, welfare, and smolt migration in hatchery-reared Atlantic salmon
  • 2017
  • Ingår i: Canadian Journal of Fisheries and Aquatic Sciences. - : Canadian Science Publishing. - 0706-652X .- 1205-7533. ; 74:3, s. 396-405
  • Tidskriftsartikel (refereegranskat)abstract
    • Hatchery-reared salmonids released into the wild generally have poor survivability compared with wild conspecifics. To assess potential hatchery rearing improvements, behavioral and physiological effects of reducing animal density and adding in-tank shelter were investigated. Atlantic salmon (Salmo salar) parr were placed in barren or shelter-enriched tanks at high or low density up until release as smolts. Lowered density rendered positive effects on growth and intestinal barrier function, and both lowered density and shelter decreased conspecific aggression, as inferred by fin damage. Furthermore, while the presence of shelter decreased stress hormone levels following human disturbance, it also decreased growth and smolt migration success, an effect particularly pronounced at high densities. Therefore, we suggest that this type of structural enrichment should be avoided for Atlantic salmon smolts held at high densities and conclude that a lowered animal density with or without shelter has the highest potential in producing a more resilient smolt for stocking.
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5.
  • Rosengren, Malin, 1982, et al. (författare)
  • High risk no gain-metabolic performance of hatchery reared Atlantic salmon smolts, effects of nest emergence time, hypoxia avoidance behaviour and size
  • 2017
  • Ingår i: Physiology & Behavior. - : Elsevier BV. - 0031-9384 .- 1873-507X. ; 175, s. 104-112
  • Tidskriftsartikel (refereegranskat)abstract
    • When animals are reared for conservational releases it is paramount to avoid reducing genetic and phenotypic variation over time. This requires an understanding of how diverging behavioural and physiological traits affect performance both in captivity and after release. In Atlantic salmon, emergence time from the spawning gravel has been linked to certain behavioural and physiological characteristics and to the concept of stress coping styles. Early emerging fry has for example been shown to be bolder and more aggressive and to have higher standard metabolic rates compared to late emerging fry. The first aim was therefore to examine if emergence latency affect the behavioural stress coping response also beyond the fry and parr stage. This was done using a hypoxia avoidance test, where an active behavioural avoidance response can be related to higher risk taking. No behavioural differences were found between the two emergence fractions either at the parr or pre-smolt stage, instead smaller individuals were more prone to express an "active" hypoxia avoidance response. Further, an individual expressing a "passive" response as parr were also more prone to express this behaviour at the pre-smolt stage. While there are some previous studies showing that early emerging individuals with a bolder personality may be favored within a hatchery setting it is not known to what extent these early differences persist to affect performance after release. The second aim was therefore to compare the physiological performance at the time of release as smolts using the two subgroups; 1) early emerging fish showing active hypoxia avoidance (Early + Bold) and 2) late emerging fish showing a passive hypoxia response (Late + Shy). The Early + Bold group showed a higher red blood cell swelling, suggesting a higher adrenergic output during stress, whereas there was no difference in post-stress plasma cortisol or physiological smolt status. While there was no difference in standard metabolic rate between the groups, the Early + Bold group exhibited a lower maximum metabolic rate and aerobic scope following strenuous swimming. In captivity this may have no clear negative effects, but in the wild, a more risk prone behavioural profile linked to a lower aerobic capacity to escape from e.g. a predator attack, could clearly be disadvantageous.
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6.
  • Zeng, Jiao, et al. (författare)
  • High-resolution structure of a fish aquaporin reveals a novel extracellular fold
  • 2022
  • Ingår i: Life Science Alliance. - : Life Science Alliance, LLC. - 2575-1077. ; 5:12
  • Tidskriftsartikel (refereegranskat)abstract
    • Aquaporins are protein channels embedded in the lipid bilayer in cells from all organisms on earth that are crucial for water homeostasis. In fish, aquaporins are believed to be important for osmoregulation; however, the molecular mechanism behind this is poorly understood. Here, we present the first structural and functional characterization of a fish aquaporin; cpAQP1aa from the fresh water fish climbing perch (Anabas testudineus), a species that is of high osmoregulatory interest because of its ability to spend time in seawater and on land. These studies show that cpAQP1aa is a water-specific aquaporin with a unique fold on the extracellular side that results in a constriction region. Functional analysis combined with molecular dynamic simulations suggests that phosphorylation at two sites causes structural perturbations in this region that may have implications for channel gating from the extracellular side.
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7.
  • Abro, Rani, et al. (författare)
  • Evaluation of chitinolytic activities and membrane integrity in gut tissues of Arctic charr (Salvelinus alpinus) fed fish meal and zygomycete biomass.
  • 2014
  • Ingår i: Comparative Biochemistry and Physiology - Part B: Biochemistry & Molecular Biology. - : Elsevier BV. - 1096-4959 .- 1879-1107. ; 175, s. 1-8
  • Tidskriftsartikel (refereegranskat)abstract
    • Chitinolytic activity, nutrient uptake and intestinal barrier functions were investigated in gut tissues of Arctic charr (Salvelinus alpinus) fed iso-nitrogenous diets based on fish meal, with or without inclusion of zygomycetes (Rhizopus oryzae). We found that gut tissue of Arctic charr had significant chitinase activity, of both endo- and exo-chitinase iso-forms. Moreover, the distribution pattern along the GI tract of Arctic charr differed between endo-chitinase and exo-chitinase. The endo-chitinase activity in stomach tissue and in the distal intestine was several hundred-fold higher than the exo-chitinase activity in stomach tissue. The greatest exo-chitinase activity was found in the distal intestine. The zygomycete-based diet resulted in higher chitinolytic activity in gut tissue compared to the fish meal-based diet. Disturbed intestinal integrity and increased uptake rate of the amino acid lysine were observed in the distal, but not proximal, intestine of fish fed the zygomycete-based feed.
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8.
  • Albertsson, Eva, 1979, et al. (författare)
  • Marin fiskodling på den svenska västkusten: Biologiska förutsättningar : Rapport nummer 1 från Vattenbrukscentrum Väst, Göteborgs universitet
  • 2013
  • Rapport (övrigt vetenskapligt/konstnärligt)abstract
    • I den av regeringen beställda utredningen ”Det växande Vattenbrukslandet” föreslås att ”Göteborgs universitet (GU) tillförs resurser för att fortsätta att utveckla vattenbruksrelaterad forskning när det gäller musslor, ostron och marina fiskarter”. Därför tog under 2011 företrädare inom den odlingsrelaterade fiskforskningen vid Gö- teborgs universitet initiativet till denna studie om förutsättningarna för marin fiskodling. Studien har syftet att utreda de odlingsbiologiska förutsättningarna för etablering av vattenbruk av marina fiskarter på den svenska västkusten. Målet är att den ska kunna användas som kunskapsunderlag för en framtida handlingsplan för fiskodling på västkusten, och främja kunskapsutbytet mellan den fiskodlingsrelaterade forskningen, beslutsfat- tande organ och privata entreprenörer för att utveckla konkurrenskraftiga innovationer inom vattenbruket. En central fråga för sammanhanget är vilka fiskarter det finns potential för att odla på den svenska västkusten. Vattenbruket måste vara ekonomiskt bärkraftigt och en urvalsmodell baserad på svenska marina fiskarters kg- pris i förhållande till en minsta tänkbara produktionskostnad användes därför i denna studie. Vidare bedöm- des arterna utifrån kunskapsläge och odlingspotential. Arterna hälleflundra, tunga, piggvar, havskatt (vanlig och fläckig) samt lyrtorsk och torsk bedömdes att vara ”kandidatarter” för framtida odling i Sverige. För dessa arter gjordes omfattande studier för att vidare utreda behov och möjligheter. Det gjordes även en analys av de abiotiska faktorerna (vattentemperatur, syrgas- och salthalt) på den svenska västkusten. I en sista del av studien kombinerades kunskapen om kandidatarternas biologiska förutsättningar med västkustens abiotiska faktorer. De abiotiska förhållandena på den svenska västkusten är inte odlingsbiologiskt optimala för traditionell nät- kasseodling för någon av de framtagna kandidatarterna, då för höga sommartemperaturer skapar problem för kallvattensarterna medan varmvattensarterna hämmas av de låga vintertemperaturerna. En av de huvudsak- liga slutsatserna i denna rapport är därför att året-runt-bedriven fiskodling i öppna system, såsom i kustnära nätkassar, knappast är praktiskt genomförbart på den svenska västkusten för de arter som i denna studie i övrigt ansågs ha hög odlingspotential. För etablering av framgångsrika odlingssystem behövs speciella tekniska lösningar, som möjliggör reglering av abiotiska faktorer såsom syrgashalter och temperatur. För att Sverige skall kunna bli ett växande vattenbruksland krävs samordnade insatser mellan forskare från flera discipliner, privata aktörer och andra intressenter. Dessutom krävs satsningar på forskning och utredning av de mera tekniskt avancerade odlingssystem som den svenska västkusten kommer behöva. Med tanke på miljöbelastningsproblematiken kring öppna havsbaserade system för fiskodling samt de abiotiska faktorerna längs västkusten som inte är optimala för någon av kandidatarterna i utredningen, så är slutsatsen att man bör undersöka, alternativa odlingssystem till traditionella öppna nätkassar för fiskodling, där man kan kontrollera faktorer som temperatur, salthalt och syrenivåer och minimera utsläppen. Nästa steg som föreslås är att göra en teknisk/ekonomisk utredning kring hållbara havs- eller landbaserade system för marin fiskodling. Utred- ningen bör innehålla fall-studier för en eller två fiskarter och ett kräftdjur och inkludera en myndighetspröv- ning för de framtagna koncepten. Ytterligare en förutsättning för att vattenbruket skall kunna utvecklas är satsningar på nödvändig infrastruktur, som ett marint kläckeri för forskning och utvecklingsarbete.
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9.
  • Alves, R. N., et al. (författare)
  • Structural and functional maturation of skin during metamorphosis in the Atlantic halibut (Hippoglossus hippoglossus)
  • 2018
  • Ingår i: Cell and Tissue Research. - : Springer Science and Business Media LLC. - 0302-766X .- 1432-0878. ; 372:3, s. 469-492
  • Tidskriftsartikel (refereegranskat)abstract
    • To establish if the developmental changes in the primary barrier and osmoregulatory capacity of Atlantic halibut skin are modified during metamorphosis, histological, histochemical, gene expression and electrophysiological measurements were made. The morphology of the ocular and abocular skin started to diverge during the metamorphic climax and ocular skin appeared thicker and more stratified. Neutral mucins were the main glycoproteins produced by the goblet cells in skin during metamorphosis. Moreover, the number of goblet cells producing neutral mucins increased during metamorphosis and asymmetry in their abundance was observed between ocular and abocular skin. The increase in goblet cell number and their asymmetric abundance in skin was concomitant with the period that thyroid hormones (THs) increase and suggests that they may be under the control of these hormones. Several mucin transcripts were identified in metamorphosing halibut transcriptomes and Muc18 and Muc5AC were characteristic of the body skin. Na+, K+-ATPase positive (NKA) cells were observed in skin of all metamorphic stages but their number significantly decreased with the onset of metamorphosis. No asymmetry was observed between ocular and abocular skin in NKA cells. The morphological changes observed were linked to modified skin barrier function as revealed by modifications in its electrophysiological properties. However, the maturation of the skin functional characteristics preceded structural maturation and occurred at stage 8 prior to the metamorphic climax. Treatment of Atlantic halibut with the THs disrupter methimazole (MMI) affected the number of goblet cells producing neutral mucins and the NKA cells. The present study reveals that the asymmetric development of the skin in Atlantic halibut is TH sensitive and is associated with metamorphosis and that this barrier's functional properties mature earlier and are independent of metamorphosis.
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10.
  • Andersson, Marica, et al. (författare)
  • Low Holding Densities Increase Stress Response and Aggression in Zebrafish
  • 2022
  • Ingår i: Biology. - : MDPI AG. - 2079-7737. ; 11:5
  • Tidskriftsartikel (refereegranskat)abstract
    • Simple Summary Zebrafish are used as experimental animals in labs all around the world. To ensure that the health of zebrafish is maintained at the highest level, it is important to know the optimal housing conditions of the animals, including the housing density. Guidelines for housing densities of zebrafish can then be spread and followed globally, making it possible to compare research data from different facilities. To investigate the optimal housing densities of zebrafish and to better understand how holding density affects zebrafish behaviour and physiology, we evaluated the welfare of zebrafish housed at different densities for nine weeks. We observed that fish housed at the lowest holding density of 1 fish/L stood out from the rest of the experimental fish, showing higher levels of aggression, secreting more of the stress hormone cortisol in the water, and spending more time in the top zone of the tank, possibly reflecting the fact that fish in this density were hiding more behind the floating plants. Our data indicate that zebrafish should not be kept at 1 fish/L, or lower, to ensure good welfare of the animals. With laboratory zebrafish (Danio rerio) being an established and popular research model, there is a need for universal, research-based husbandry guidelines for this species, since guidelines can help promote good welfare through providing appropriate care. Despite the widespread use of zebrafish in research, it remains unclear how holding densities affect their welfare. Previous studies have mainly evaluated the effects of holding densities on a single parameter, such as growth, reproductive output, or social interactions, rather than looking at multiple welfare parameters simultaneously. Here we investigated how chronic (nine weeks) exposure to five different holding densities (1, 4, 8, 12, and 16 fish/L) affected multiple welfare indicators. We found that fish in the 1 fish/L density treatment had higher free water cortisol concentrations per fish, increased vertical distribution, and displayed aggressive behaviour more frequently than fish held at higher densities. On the other hand, density treatments had no effect on anxiety behaviour, whole-brain neurotransmitter levels, egg volume, or the proportion of fertilised eggs. Our results demonstrate that zebrafish can be held at densities between 4 and 16 fish/L without compromising their welfare. However, housing zebrafish in the density of 1 fish/L increased their stress level and aggressive behaviour.
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