Temporal fluctuation of thiamine deficiency is a well-documented phenomenon in Atlantic salmon (Salmo salar) from the Baltic Sea. The deficiency causes increased levels of fry mortality (M74-syndrome). Although the cause for the deficiency in salmon remains somewhat unclear, the mortality rate can be significantly reduced by bathing alevins in thiamine solution. However, it is not known whether thiamine supplementation at the alevin stage successfully treats all other potential adverse effects associated to thiamine deficiency during the embryonic stages of development. To investigate how thiamine deficiency affects growth and survival in thiamine treated and untreated salmon during the first summer, fertilized eggs with known thiamine content were sorted into four groups: Very low, Low, Medium and High thiamine content (all having thiamine levels above critical levels where mortality is inevitable if left untreated). Half of each group was treated with thiamine according to standardized hatchery procedures and half was left untreated as control. Mortality was monitored continuously, and body length and -mass were recorded after the initial growing season (five months after hatching). The results showed that fish with very low initial levels of thiamine grew smaller even if treated with supplemental thiamine. This indicates a potential disturbance of individuals suffering from early thiamine deficiency.

Fishing has the potential to influence the life-history traits of exploited populations. However, our understanding of how fisheries can induce evolutionary genetic changes remains incomplete. The discovery of large-effect loci linked with ecologically important life-history traits, such as age at maturity in Atlantic salmon (Salmo salar), provides an opportunity to study the impacts of temporally varying fishing pressures on these traits. A 93-year archive of fish scales from wild Atlantic salmon catches from the northern Baltic Sea region allowed us to monitor variation in adaptive genetic diversity linked with age at maturity of wild Atlantic salmon populations. The dataset consisted of samples from both commercial and recreational fisheries that target salmon on their spawning migration. Using a genotyping-by-sequencing approach (GT-seq), we discovered strong within-season allele frequency changes at the vgll3 locus linked with Atlantic salmon age at maturity: fishing in the early season preferentially targeted the vgll3 variant linked with older maturation. We also found within-season temporal variation in catch proportions of different wild Atlantic salmon subpopulations. Therefore, selective pressures of harvesting may vary depending on the seasonal timing of fishing, which has the potential to cause evolutionary changes in key life-history traits and their diversity. This knowledge can be used to guide fisheries management to reduce the effects of fishing practices on salmon life-history diversity. Thus, this study provides a tangible example of using genomic approaches to infer, monitor and help mitigate human impacts on adaptively important genetic variation in nature.

Throughout their native range, adfluvial brown trout populations have declined due to habitat degradation and over exploitation. As a consequence, numerous restoration projects that include stocking of hatchery-reared trout have been conducted; however, estimates of survival in large lakes remain scarce. Here, we use telemetry to evaluate the survival of stocked trout smolts and sub-adults in a large lake in central Sweden. Of the smolts released in the main tributary, 42% entered the lake. Both life stages suffered high rates of mortality. Only 8% of smolts survived the first 10 months after entering the lake and 10% of those released as sub-adults survived 23 months post-release. In agreement with studies on anadromous trout, we conclude that stocking of hatchery-reared individuals is not an efficient method to recover population numbers due to poor performance in the wild. Similar studies from other large lakes and comparisons with wild conspecifics would increase the applicability of the results and promote further understanding of the utilization of hatchery-origin trout to rebuild wild populations and sustain fisheries.

Evolution of iteroparity is shaped by the trade-off between current and future reproduction. We studied variation in iteropar- ity among 205050 individual Atlantic salmon caught in 179 rivers spanning 14 degrees of latitude. The proportion of repeat spawners (iteroparous individuals) averaged 3.8% and ranged from 0% to 26% across rivers. Females were more often repeat spawners than males and had lower cost of reproduction in terms of lost body mass between spawning events. Proportion of repeat spawners for a given sea age at maturity, and the ratio of alternate to consecutive repeat spawners, increased with increasing population mean sea age at maturity. By combining smolt age, sea age at maturity, and age at additional spawning events, we identified 141 unique life-history types, and repeat spawners contributed 75% of that variation. Our results show that repeat spawners are important for life-history variation and suggest that the association between mean sea age and the frequency of repeat spawning is adaptive rather than a pleiotropic side effect arising from selection on sea age.

Evolution of phenotypic plasticity requires genotype-environment interaction. The discovery of two large-effect loci in the vgll3 and six6 genomic regions associated with the number of years the Atlantic salmon spend feeding at sea before maturation (sea age), provides a unique opportunity to study evolutionary potential of phenotypic plasticity. Using data on 1246 Atlantic salmon caught in the River Surna in Norway, we show that variation in mean sea age among years (smolt cohorts 2013-2018) is influenced by genotype frequencies as well as interaction effects between genotype and year. Genotype-year interactions suggest that genotypes may differ in their response to environmental variation across years, implying genetic variation in phenotypic plasticity. Our results also imply that plasticity in sea age will evolve as an indirect response to selection on mean sea age due to a shared genetic basis. Furthermore, we demonstrate differences between years in the additive and dominance functional genetic effects of vgll3 and six6 on sea age, suggesting that evolutionary responses will vary across environments. Considering the importance of age at maturity for survival and reproduction, genotype-environment interactions likely play an important role in local adaptation and population demography in Atlantic salmon.

Anadromous brown trout ( Salmo trutta ) in the northeastern Atlantic Ocean, and coastal cutthroat trout ( Oncorhynchus clarkii clarkii ) in the northeastern Pacific Ocean, are analogs in many ways. Both species display similar patterns of complex life histories and behavioral flexibility, are iteroparous and facultatively anadromous and occupy nearshore coastal marine habitats where numerous populations often mix. These characteristics create specific challenges for management and conservation that have been complicated by inadequate scientific attention in some areas. Both species are declining across their native range, and their ecology make them particularly vulnerable to habitat destruction, fishing, and climate change. Here, we review the available literature to compare the biology, ocean ecology, and management practices of these two species. We highlight ecological similarities and differences between the species and identify current knowledge gaps suggesting future research needs and management actions. Using a comparative approach, the review aims to promote and facilitate knowledge exchange between anadromous trout research communities from opposite sides of the globe to improve management and conservation of these species and stimulate the production of management plans specific to anadromous trout.

Pharmaceutical pollution represents a rapidly growing threat to ecosystems worldwide. Drugs are now commonly detected in the tissues of wildlife and have the potential to alter the natural expression of behavior, though relatively little is known about how pharmaceuticals impact predator-prey interactions. We conducted parallel laboratory experiments using larval odonates (dragonfly and damselfly nymphs) to investigate the effects of exposure to two pharmaceuticals, cetirizine and citalopram, and their mixture on the outcomes of predator-prey interactions. We found that exposure to both compounds elevated dragonfly activity and impacted their predation success and efficiency in complex ways. While exposure to citalopram reduced predation efficiency, exposure to cetirizine showed varied effects, with predation success being enhanced in some contexts but impaired in others. Our findings underscore the importance of evaluating pharmaceutical effects under multiple contexts and indicate that these compounds can affect predator-prey outcomes at sublethal concentrations.

Recent and exciting in vivo studies show that supplementation with the polyamine spermidine (Spd) is cardioprotective and prolongs lifespan in both mice and humans. The mechanisms behind Spd-induced cardioprotection are supposed to involve Spd-evoked stimulation of autophagy, mitophagy and mitochondrial respiration and improved the mechano-elastical function of cardiomyocytes. Although cellular uptake of Spd was not characterized, these results suggest that Spd is imported by the cardiomyocytes and acts intracellularly. In the light of these new and thrilling data, we discuss in the present review cellular polyamine import with a special focus on mechanisms that may be relevant for Spd uptake by electrically excitable cells such as cardiomyocytes.

Polyamines are small aliphatic cationic molecules synthesized via a highly regulated pathway and involved in general molecular and cellular phenomena. Both mammalian cells and microorganisms synthesize polyamines, and both sources may contribute to the presence of polyamines in the circulation. The dominant location for microorganisms within the body is the gut. Accordingly, the gut microbiota probably synthesizes most of the polyamines in the circulation in addition to those produced by the mammalian host cells. Polyamines are mandatory for cellular growth and proliferation. Established evidence suggests that the polyamine spermidine prolongs lifespan and improves cardiovascular health in animal models and humans through both local mechanisms, involving improved cardiomyocyte function, and systemic mechanisms, including increased NO bioavailability and reduced systemic inflammation. Higher levels of polyamines have been detected in non-dilated aorta of patients affected by bicuspid aortic valve congenital malformation, an aortopathy associated with an increased risk for thoracic ascending aorta aneurysm. In this review, we discuss metabolism of polyamines and their potential effects on vascular smooth muscle and endothelial cell function in vascular pathology of the thoracic ascending aorta associated with bicuspid or tricuspid aortic valve.

This paper reviews the life history of brown trout and factors influencing decisions to migrate. Decisions that maximize fitness appear dependent on size at age. In partly anadromous populations, individuals that attain maturity at the parr stage typically become freshwater resident. For individual fish, the life history is not genetically fixed and can be modified by the previous growth history and energetic state in early life. This phenotypic plasticity may be influenced by epigenetic modifications of the genome. Thus, factors influencing survival and growth determine life-history decisions. These are intra- and interspecific competition, feeding and shelter opportunities in freshwater and salt water, temperature in alternative habitats and flow conditions in running water. Male trout exhibit alternative mating strategies and can spawn as a subordinate sneaker or a dominant competitor. Females do not exhibit alternative mating behaviour. The relationship between growth, size and reproductive success differs between sexes in that females exhibit a higher tendency to migrate than males. Southern populations are sensitive to global warming. In addition, fisheries, aquaculture with increased spreading of salmon lice, introduction of new species, weirs and river regulation, poor water quality and coastal developments all threaten trout populations. The paper summarizes life-history data from six populations across Europe and ends by presenting new research questions and directions for future research.

Fish display a remarkable diversity in juvenile migration strategies and behavior. Intra-species variation in migration can be considerable, and understanding the driving force of such variation is important for effective management and conservation of migratory fish. In facultative migratory species, such as many salmonid fish, energetic status is known to affect migration strategy and behavior. However, we currently lack a full understanding of how energetic status affects juvenile development and migration over different environmental contexts. In this study, we examined the effect of energetic status on juvenile migration initiation and migratory behaviors in 1 and 2 year old brown trout (Salmo trutta). By manipulating feeding regimes, we created a large variation in trout energetic status (using condition factor as a proxy). We then studied behavioral changes in migration in both a controlled environment (large-scale migration pools) as well as a natural river system using both passive integrative transponder tags (PIT-tags) as well as acoustic telemetry tags. In the laboratory setting, 1 year old trout with higher energetic status were more likely to initiate migration and migrated faster. For 2 year old trout, energetic status did not affect the initiation of migration (the large majority migrated), but high energetic status fish migrated faster. In a small-scale natural creek system, few age one fish migrated (11%); however, these few migrators were within the upper range of energetic status. In 2 year old trout, a high percentage became migrants (79%), and those with higher energetic status migrated at a faster speed. In a large-scale river system, successful downstream seaward migration for 2 year olds was low (9%) and independent of energetic status. Our findings provide valuable data for fisheries management because we show that age at release and energetic status prior to release can impact migration initiation and behaviors. Our findings also indicate that migration measured in the laboratory may over estimate migration in the wild, especially for younger, age one fish. More broadly, this work advances our understanding of this complex life history stage and the mechanisms involved in the initiation, behavior, and survival of migrating brown trout.

Hydropower development has negatively influenced Atlantic salmon (Salmo salar) populations. Compensatory hatchery‐rearing programmes exist, but released fish suffer from high mortality that may be related to the lack of experience from natural environments in hatchery‐reared smolts and their large body size and high energetic state. Here, we used acoustic telemetry to test how body size, energetic state, and the environmental conditions of the river affect migration in hatchery‐reared smolts. The study was conducted in three consecutive years between 2011 and 2013 in the lower part of the River Umealven, Sweden. For individual fish, there was no effect of body size but the energetic state of the fish had a negative effect on sea entry. The most important factor affecting sea entry rate was the water discharge in the old river bed that differed among years. Smolts were more likely to enter the sea in years when the discharge was high or when the discharge increased substantially shortly after release. Hatchery‐reared fish had higher migration speed at a slower flowing section compared with a faster flowing section, which was likely a result of large hesitation to enter the rapid section. The increase in water discharge led to an increase in fish migration speed disproportional to the increase in water velocity. Our results highlight the importance of water discharge for the smolts during smolt migration, and we argue that concern should be given to migrating fish when managing regulated rivers.

Tumors are phenotypically heterogeneous and include subpopulations of cancer cells with stemlike properties. The natural product salinomycin, a K+-selective ionophore, was recently found to exert selectivity against such cancer stem cells. This selective effect is thought to be due to inhibition of the Wnt signaling pathway, but the mechanistic basis remains unclear. Here, we develop a functionally competent fluorescent conjugate of salinomycin to investigate the molecular mechanism of this compound. By subcellular imaging, we demonstrate a rapid cellular uptake of the conjugate and accumulation in the endoplasmic reticulum (ER). This localization is connected to induction of Ca2+ release from the ER into the cytosol. Depletion of Ca2+ from the ER induces the unfolded protein response as shown by global mRNA analysis and Western blot analysis of proteins in the pathway. In particular, salinomycin-induced ER Ca2+ depletion up-regulates C/EBP homologous protein (CHOP), which inhibits Wnt signaling by down-regulating β-catenin. The increased cytosolic Ca2+ also activates protein kinase C, which has been shown to inhibit Wnt signaling. These results reveal that salinomycin acts in the ER membrane of breast cancer cells to cause enhanced Ca2+ release into the cytosol, presumably by mediating a counter-flux of K+ ions. The clarified mechanistic picture highlights the importance of ion fluxes in the ER as an entry to inducing phenotypic effects and should facilitate rational development of cancer treatments.

Polyamines are cationic molecules synthesized via a highly regulated pathway, obtained from the diet or produced by the gut microbiota. They are involved in general molecular and cellular phenomena that play a role also in vascular disease. Bicuspid aortic valve (BAV) is a congenital malformation associated to a greater risk of thoracic ascending aorta (TAA) aneurysm, whose pathogenesis is not yet well understood. We focused on differential analysis of key members of polyamine pathway and on polyamine concentration in non-dilated TAA samples from patients with either stenotic tricuspid aortic valve (TAV) or BAV (diameter ≤ 45 mm), vs. normal aortas from organ donors, with the aim of revealing a potential involvement of polyamines in early aortopathy. Changes of gene expression in TAA samples were evaluated by RT-PCR. Changes of ornithine decarboxylase 1 (ODC1), a key enzyme in polyamine formation, and cationic amino acid transporter 1 (SLC7A1/CAT-1) expression were analyzed also by Western blot. ODC1 subcellular localization was assessed by immunohistochemistry. Polyamine concentration in TAA samples was evaluated by HPLC. BAV TAA samples showed an increased concentration of putrescine and spermidine vs. TAV and donor samples, together with a decreased mRNA level of polyamine anabolic enzymes and of the putative polyamine transporter SLC7A1/CAT-1. The catabolic enzyme spermidine/spermine N1-acetyltransferase 1 showed a significant mRNA increase in TAV samples only, together with a decreased concentration of spermine. The decreased expression of SLC7A1/CAT-1 and ODC1 mRNAs in BAV corresponded to increased or unchanged expression of the respective proteins. ODC was located mainly in smooth muscle cell (SMC) nucleus in TAV and donor samples, while it was present also in SMC cytoplasm in BAV samples, suggesting its activation. In conclusion, BAV, but not TAV non-dilated samples show increased polyamine concentration, accompanied by the activation of a regulatory negative feedback mechanism.

The aminopropyltransferase spermidine synthase (SpdS) is a promising drug target in cancer and in protozoan diseases including malaria. Plasmodium falciparum SpdS (PfSpdS) transfers the aminopropyl group of decarboxylated S-adenosylmethionine (dcAdoMet) to putrescine or to spermidine to form spermidine or spermine, respectively. In an effort to understand why efficient inhibitors of PfSpdS have been elusive, the present study uses enzyme activity assays and isothermal titration calorimetry with verified or predicted inhibitors of PfSpdS to analyze the relationship between binding affinity as assessed by KD and inhibitory activity as assessed by IC50. The results show that some predicted inhibitors bind to the enzyme with high affinity but are poor inhibitors. Binding studies with PfSpdS substrates and products strongly support an ordered sequential mechanism in which the aminopropyl donor (dcAdoMet) site must be occupied before the aminopropyl acceptor (putrescine) site can be occupied. Analysis of the results also shows that the ordered sequential mechanism adequately accounts for the complex relationship between IC50 and KD and may explain the limited success of previous efforts at structure-based inhibitor design for PfSpdS. Based on PfSpdS active-site occupancy, we suggest a classification of ligands that can help to predict the KD-IC50 relations in future design of new inhibitors. The present findings may be relevant for other drug targets that follow an ordered sequential mechanism.

Cancer stem cells (CSCs) have been invoked in resistance, recurrence and metastasis of cancer. Consequently, curative cancer treatments may be contingent on CSC selective approaches. Of particular interest in this respect is the ionophore salinomycin, a natural product shown to be 100-fold more active against CSCs than clinically used paclitaxel. We have previously reported that synthetic salinomycin derivatives display increased activity against breast cancer cell lines. Herein we specifically investigate the CSC selectivity of the most active member in each class of C20-O-acylated analogs as well as a C1-methyl ester analog incapable of charge-neutral metal ion transport.

Migration is an important life-history event in a wide range of taxa, yet many migrations are influenced by anthropogenic change. Although migration dynamics are extensively studied, the potential effects of environmental contaminants on migratory physiology are poorly understood. In this study we show that an anxiolytic drug in water can promote downward migratory behaviour of Atlantic salmon (Salmo salar) in both laboratory setting and in a natural river tributary. Exposing salmon smolt to a dilute concentration of a GABAA receptor agonist (oxazepam) increased migration intensity compared with untreated smolt. These results implicate that salmon migration may be affected by human-induced changes in water chemical properties, such as acidification and pharmaceutical residues in wastewater effluent, via alterations in the GABAA receptor function.

Increased vascular smooth muscle cell (VSMC) proliferation is a factor in atherosclerosis and injury‐induced arterial (re) stenosis. Inhibition of polyamine synthesis by α‐difluoro‐methylornithine (DFMO), an irreversible inhibitor of ornithine decarboxylase, attenuates VSMC proliferation with high sensitivity and specificity. However, cells can escape polyamine synthesis blockade by importing polyamines from the environment. To address this issue, polyamine transport inhibitors (PTIs) have been developed. We investigated the effects of the novel trimer44NMe (PTI‐1) alone and in combination with DFMO on VSMC polyamine uptake, proliferation and phenotype regulation. PTI‐1 efficiently inhibited polyamine uptake in primary mouse aortic and human coronary VSMCs in the absence as well as in the presence of DFMO. Interestingly, culture with DFMO for 2 days substantially (>95%) reduced putrescine (Put) and spermidine (Spd) contents without any effect on proliferation. Culture with PTI‐1 alone had no effect on either polyamine levels or proliferation rate, but the combination of both treatments reduced Put and Spd levels below the detection limit and inhibited proliferation. Treatment with DFMO for a longer time period (4 days) reduced Put and Spd below their detection limits and reduced proliferation, showing that only a small pool of polyamines is needed to sustain VSMC proliferation. Inhibited proliferation by polyamine depletion was associated with maintained expression of contractile smooth marker genes. In cultured intact mouse aorta, PTI‐1 potentiated the DFMO‐induced inhibition of cell proliferation. The combination of endogenous polyamine synthesis inhibition with uptake blockade is thus a viable approach for targeting unwanted vascular cell proliferation in vivo, including vascular restenosis. J. Cell. Physiol. 231: 1334–1342, 2016. © 2015 Wiley Periodicals, Inc.

The enzymes of the polyamine-biosynthesis pathway have been proposed to be promising drug targets in the treatment of malaria. Spermidine synthase (SpdS; putrescine aminopropyltransferase) catalyzes the transfer of the aminopropyl moiety from decarboxylatedS-adenosylmethionine to putrescine, leading to the formation of spermidine and 5′-methylthioadenosine (MTA). In this work, X-ray crystallography was used to examine ligand complexes of SpdS from the malaria parasitePlasmodium falciparum(PfSpdS). Five crystal structures were determined ofPfSpdS in complex with MTA and the substrate putrescine, with MTA and spermidine, which was obtained as a result of the enzymatic reaction taking place within the crystals, with dcAdoMet and the inhibitor 4-methylaniline, with MTA and 4-aminomethylaniline, and with a compound predicted in earlierin silicoscreening to bind to the active site of the enzyme, benzimidazol-(2-yl)pentan-1-amine (BIPA). In contrast to the other inhibitors tested, the complex with BIPA was obtained without any ligand bound to the dcAdoMet-binding site of the enzyme. The complexes with the aniline compounds and BIPA revealed a new mode of ligand binding toPfSpdS. The observed binding mode of the ligands, and the interplay between the two substrate-binding sites and the flexible gatekeeper loop, can be used in the design of new approaches in the search for new inhibitors of SpdS.

The polyamines are important for a variety of cellular functions, including cell growth. Their intracellular concentrations are controlled by a complex network of regulatory mechanisms, in which antizyme (Az) has a key role. Az reduces the cellular polyamine content by down-regulating both the enzyme catalysing polyamine biosynthesis, ornithine decarboxylase (ODC), and the uptake of polyamines. The activity of Az is repressed by the binding of a protein, named Az inhibitor (AzI), which is an enzymatically inactive homologue of ODC. Two forms of AzI have been described: AzI1, which is ubiquitous, and AzI2 which is expressed in brain and testis. In the present study, we have investigated the role of AzI1 in polyamine homeostasis and cell proliferation in breast cancer cells. The results obtained showed that the cellular content of AzI increased transiently after induction of cell proliferation by diluting cells in fresh medium. Inhibition of polyamine biosynthesis induced an even larger increase in the cellular AzI content, which remained significantly elevated during the 7-day experimental period. However, this increase was not a consequence of changes in cell cycle progression, as demonstrated by flow cytometry. Instead, the increase appeared to correlate with the cellular depletion of polyamines. Moreover, induced overexpression of AzI resulted in an increased cell proliferation with a concomitant increase in ODC activity and putrescine content. During mitosis, AzI1 was localised in a pattern that resembled that of the two centrosomes, confirming earlier observations. Taken together, the results indicate that AzI fulfils an essential regulatory function in polyamine homeostasis and cell proliferation.