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Trophic interactions in the pelagic area of lakes and the opposing effects of fish feeding (top-down) and phytoplankton biomass (bottom-up) on zooplankton communities are central topics in limnology. We hypothesized that zooplankton size distributions should be a more sensitive approach to disentangle top-down and bottom-up effects than the commonly measured zooplankton biomass. We examined zooplankton size distributions from 148 samples collected during summer months in the upper and lower pelagic layers of a deep mesotrophic lake among 13 years of sampling. Top-down effects, namely fish size and biomass, and bottom-up effects, including water temperature and total phosphorus and chlorophyll a concentrations, were considered. To add robustness to our analyses, we expressed the zooplankton size distributions as size spectra based on log-binning, as continuous size spectra and by the size diversity, a measure that has been developed to mimic taxonomic diversity indices. Among numerous regressions tested, significant top-down or bottom-up effects could rarely been detected. Our results indicate that the overall zooplankton size distribution was not significantly affected by fish predation and lake productivity measured as total phosphorus or chlorophyll a concentration. However, we found negative correlations between fish biomass and the preferred zooplankton prey, including Bosmina longirostris, Daphnia cucullata and nauplii in the upper depth layer. However, due to their small body size, low biomass and therefore relative small contribution to the zooplankton size distribution, predation on preferred zooplankton species did not translate into a statistically significant modification of the entire size spectrum. Consequently, the size spectrum seems to be relatively robust against predation effects, but might reflect the lake-wide energy availability and transfer efficiency in the food web.

A major clinical challenge of ovarian cancer is the development of malignant ascites accompanied by widespread peritoneal metastasis. In ovarian clear cell carcinoma (OCCC), a challenging subtype of ovarian cancer, this problem is compounded by near-universal primary chemoresistance; patients with advanced stage OCCC thus lack effective therapies and face extremely poor survival rates. Here we show that tumor-cell-expressed serine protease inhibitor Kazal type 1 (SPINK1) is a key driver of OCCC progression and metastasis. Using cell culture models of human OCCC, we find that shRNA silencing of SPINK1 sensitizes tumor cells to anoikis and inhibits proliferation. Knockdown of SPINK1 in OCCC cells also profoundly suppresses peritoneal metastasis in mouse implantation models of human OCCC. We next identify a novel autocrine signaling axis in OCCC cells whereby tumor-cell-produced interleukin-6 (IL-6) regulates SPINK1 expression to stimulate a common protumorigenic gene expression pattern leading to anoikis resistance and proliferation of OCCC cells. We further demonstrate that this signaling pathway can be successfully interrupted with the IL-6Rα inhibitor tocilizumab, sensitizing cells to anoikis in vitro and reducing metastasis in vivo. These results suggest that clinical trials of IL-6 pathway inhibitors in OCCC may be warranted, and that SPINK1 might offer a candidate predictive biomarker in this population.

Considerable theoretical work predicts that intraspecific trait variation can have profound ecological consequences by altering species interactions. Because of their high flexibility, behavioral traits may be especially relevant in mediating how species respond to one another, thus affecting food web dynamics and ecosystem functioning. However, empirical evidence supporting this idea is limited. Here, we generated predator groups where we manipulated the composition of behavioral types within the groups to assess effects on predator growth rates, prey communities, basal resources, and ecosystem functioning in replicated outdoor ponds. Using European perch (

Vendace is commonly assumed at low risk to recruitment overfishing. This assumption has been confirmed for boreal stocks but might not apply at lower latitudes. We evaluated the risk of recruitment overfishing at the southernmost extent of vendace populations by comparing fecundity, natural mortality, and fishing mortality of two stocks differing in growth with those of a population not subject to fishing. Although in one study lake vendace grew faster owing to high lake productivity and low stock density, and had substantially higher age-specific fecundity, intense harvest before first spawning negatively affected the stock's recruitment potential by reducing spawner abundance. In contrast, in the slow-growing stock, vendace entered the fishery after first spawning, and egg production per recruit was similar to that of the population not subject to fishing. We conclude that vendace stocks characterized by fast somatic growth may be at higher risk of recruitment overfishing, which can be reduced by protecting first-time spawners.

Understanding drivers of freshwater fish assemblages is critically important for biodiversity conservation strategies, especially in rapidly developing countries, which often have environmental protections lagging behind economic development. The influences of natural and human factors in structuring fish assemblages and their relative contributions are likely to change given the increasing magnitude of human activities. To discriminate natural and human drivers of fish diversity and assemblage patterns in developing countries with rapid socio-economic development, a dataset of 908 freshwater fish species and 13 metrics including three categories of both natural (i.e., biogeographic) and human drivers (i.e., economic growth, inland fisheries) in China were analysed with machine learning algorithms (i.e., self-organizing map, random forest). Here, we found that biogeographic drivers explained 21.8% of the observed fish assemblage patterns in China and remained stronger predictors when compared to human drivers (i.e., 15.6%, respectively). Freshwater fish species richness was positively correlated to rainfall, air temperature, surface water area and inland fisheries production but negatively correlated with urbanization. In addition, the strong structuring effects of climatic variables on Chinese fish richness patterns suggested that the fish assemblages could be particularly vulnerable to climate change. Our results showed that natural biogeographic factors still dominate in driving freshwater fish assemblage patterns despite increased human disturbances on aquatic ecosystems in a rapidly developing country. These findings consequently suggested that we should consider both natural (e.g., climate) and human (e.g., urbanization, inland fisheries) factors when establishing aquatic conservation strategies and priorities for developing countries that are experiencing rapid socio-economic changes.

We present a technique for the simultaneous photostructuring of hydrogels on the μm scale with different compositions on one substrate. The existing methods (e.g. spin-coating of the hydrogel precursor and application of simple moulds or spacers) have less potential to integrate spatially separated functional hydrogel dots as needed for the fabrication of complex polydimethylsiloxane (PDMS) on-glass microfluidic chips. Here, we report a method for the fast production of precisely placed hydrogel structures with adjustable height (≤80 μm) and width (≤500 μm) on glass substrates utilizing black polyoxymethylene moulds. The integration of several chambers for different hydrogel precursor solutions in one mould allows the simultaneous photopolymerization of hydrogels with different compositions. To visualize the difference in composition of the hydrogel dots and to demonstrate the precision in structuring, various quantum dots were added to the hydrogel precursor solutions and physically entrapped during the photopolymerization. To show the high potential of our photostructuring method, a microfluidic double-chamber reactor was designed and the variability in the performance of tri-enzymatic cascade reactions with optionally spatially separated or non-separated enzymes β-galactosidase, glucose oxidase, and horseradish peroxidase was visualized.

Triggered by the adoption of the Water Framework Directive, a variety of fish-based systems were developed throughout Europe to assess the ecological status of lakes. This paper provides a comprehensive overview of all existing systems and summarizes sampling methods, fish community traits (metrics) and the relevant anthropogenic pressures assessed by them. Twenty-one European countries developed fish-based assessment systems. Three countries each developed two distinct systems to approach different ecoregions, either to use different data, or to assess different lake types leading to a total number of 24 systems. The most common approach for the setting of reference conditions, used in seventeen systems, was the utilisation of fish communities in comparably undisturbed natural lakes as reference. Eleven used expert judgment, nine historical data and eight modelled relationships. Fourteen systems combined at least two approaches. The most common fish sampling method was a standardized fishing procedure with multimesh-gillnets. Many countries applied combinations of fishing methods, e.g. non-standard gillnets, fyke nets and electrofishing. Altogether 177 metrics were used for index development and each system combined 2–13 metrics. The most common ones were total standardized catches of number and biomass, relative abundance of Perca fluviatilis, Rutilus rutilus, and Abramis brama, feeding preferences, sensitive species, and non-natives. The pressure-response-relationships for these metrics were supported with both correlations established during system development and scientific publications. However, the metrics and their combinations were highly diverse and no metric was applied universally. Our analysis reveals that most fish-based assessment systems address multiple pressures (eutrophication, hydromorphological alterations, fishery pressure and occurrence of non-natives), whilst few are pressure-specific, tackling only eutrophication or acidification. We argue that the value of fish-based systems for lakes lies in their capacity to capture the effect of many different pressures and their interactions which is lacking for most assessment systems based on other biota.

Background Sympatric speciation along ecological gradients has been studied repeatedly, in particular in freshwater fishes. Rapid post-glacial ecological divergence has resulted in numerous endemic species or ecologically distinct populations in lakes of the temperate zones. Here, we focus on the Baltic cisco (Coregonus albula) complex, to study the genetic similarity among two pairs of sympatric autumn- and spring-spawning populations from post-glacial German Lakes Stechlin and Breiter Luzin. For comparison, we included a similar pair of sympatric populations from the Swedish Lake Fegen. We wanted to explore potential genetic similarities between the three sympatric cisco population pairs in the three lakes, to evaluate whether the pairs may have emerged independently in the three lakes, or whether two different species may have colonized all three lakes independently. Furthermore, we considered allopatric C. albula populations from three Polish, three Finnish, and four Swedish locations, added one Siberian population of the sister species C. sardinella and a Swedish C. maraena (whitefish) population. By genotyping nine microsatellite markers in 655 individuals from these 18 populations, we wanted to elucidate how strongly the cisco populations differ across a larger geographical area within Europe. Finally, we compared the genetic differences between the spring- and autumn-spawning populations of ciscoes in the two German lakes to infer the potentially deteriorating effect of strong anthropogenic pressure on the lakes. Results Dendrogram, Principal Coordinate Analysis and admixture analysis all indicated strong correspondence between population differentiation and geographical location for most cisco populations in Europe, including the Siberian population of C. sardinella. However, populations from some Swedish lakes deviated from this general pattern, by showing a distinct genetic structure. We found evidence for independent evolution of the three sympatric population pairs, because the populations co-occurring in the same lake were always most closely related. However, genetic differentiation was weak in the two German population pairs, but strong in the Swedish Lake Fegen, indicating that the weak differentiation in the German pairs reported earlier has eroded further. Conclusions Our results suggest that the genetic differentiation at neutral genetic markers among populations of the Baltic cisco complex has evolved (and is maintained) by random genetic drift in isolated populations. However, earlier studies on the Swedish populations combining mitochondrial DNA and microsatellite data indicate that also post-glacial immigration from separate glacial refugia has shaped the present genetic population structure. The low neutral differentiation of the German sympatric pairs in contrast to the Swedish pair suggests that recent anthropogenic effects on the lakes in Germany may put the endemic spring-spawners at risk to extinction.

Aim: We tested whether there is a strong effect of species interactions on assembly of local lake fish communities, in addition to environmental filters and dispersal. Location: Seven hundred and seventy-two European lakes and reservoirs. Time period: 1993–2012. Major taxa studied: Nineteen species of freshwater fishes. Methods: We applied a latent variable approach using Bayesian Markov chain Monte Carlo algorithms (R package “BORAL”). We compared the contributions of six environmental predictors and the spatial organization of 772 European lakes in 209 river basins on the presence/absence of the 19 most frequent fish species and on the biomass and mean mass of the six dominant species. We inspected the residual correlation matrix for positive and negative correlations between species. Results: Environmental (50%) and spatial (10%) predictors contributed to the presence/absence assembly of lake fish communities, whereas lake size and productivity contributed strongly to the biomass and mean mass structures. We found highly significant negative correlations between predator and prey fish species pairs in the presence/absence, biomass and mean mass datasets. There were more significantly positive than negative correlations between species pairs in all three datasets. In addition, unmeasured abiotic predictors might explain some of the correlations between species. Main conclusions: Strong effects of species interactions on assembly of lake fish communities are very likely. We admit that our approach is of a correlational nature and does not generate mechanistic evidence that interactions strongly shape fish community structures; however, the results fit with present knowledge about the interactions between the most frequent fish species in European lakes and they support the assumption that, in particular, the mean masses of fish species in lakes are modified by species interactions. Türkiye Bilimsel ve Teknolojik Araştirma Kurumu, Grant/Award Number: 118C250; Spanish Ministry of Science, Innovation and Universities, Grant/ Award Number: RTI2018-095363-B-I00; European Commission; Environmental Investment Centre KIK, Grant/Award Number: 8603/2014; Deutsche Forschungsgemeinschaft, Grant/Award Number: Me 1686/7-1 Türkiye Bilimsel ve Teknolojik Araştirma Kurumu, Grant/Award Number: 118C250; Spanish Ministry of Science, Innovation and Universities, Grant/ Award Number: RTI2018-095363-B-I00; European Commission; Environmental Investment Centre KIK, Grant/Award Number: 8603/2014; Deutsche Forschungsgemeinschaft, Grant/Award Number: Me 1686/7-1

Although the ray-finned fishes are named for their bony, segmented lepidotrichia (fin rays), we are only beginning to understand the morphological and functional diversity of this key vertebrate structure. Fin rays support the fin web, and their material properties help define the function of the entire fin. Many earlier studies of fin ray morphology and function have focused on isolated rays, or on rays from only one or two fins. At the same time, relatively little is known about how different preservation techniques affect the material properties of many vertebrate structures, including fin rays. Here, we use three-point bending tests to examine intra- and inter-fin variation in the flexural stiffness of fin rays from yellow perch, Perca flavescens. We sampled fin rays from individuals that were assigned to one of three preservation treatments: fresh, frozen, and preserved with formalin. The flexural stiffness of the fin rays varied within and among fins. Pelvic-fin rays were the stiffest, and pectoral fin rays the least stiff. The fin rays of the dorsal, anal, and caudal fins all had similar stiffness values, which were intermediate relative to those from the paired fins. The flexural stiffness of the fin rays was higher in rays that were at the leading edge of the fin. This variation in flexural stiffness was associated with variation in joint density and the relative length of the unsegmented proximal base of the fin rays. There was no significant difference in flexural stiffness between fresh and frozen specimens. In specimens preserved with formalin, there is a small but significant effect on stiffness in smaller fin rays.

In ectotherms, anthropogenic warming often increases energy requirements for metabolism, which can either impair growth (when resources are limiting) or lead to higher predator feeding rates and possibly stronger top–down trophic interactions. However, the relative importance of these effects in nature remains unclear because: (1) thermal adaptation or acclimation could lower metabolic costs; (2) greater prey production at warmer temperatures could compensate for higher predator feeding rates; and/or (3) temperature effects on trophic interactions via altered biological rates could be small relative to other, temperature‐unrelated human impacts on food webs. Here, we examined effects of deforestation‐associated warming on the minnow Enteromius neumayeri, occurring in both forested (cool) and deforested (warm) streams located inside or nearby an afrotropical rainforest. Combining approaches from physiological and community ecology, we quantified impacts of anthropogenic warming on the metabolism, growth and trophic interactions of this tropical ectotherm. We then compared these effects with impacts of land use unrelated to temperature. In a long‐term laboratory acclimation experiment quantifying the temperature dependence of growth and metabolism in E. neumayeri, warming increased metabolic rates and decreased growth (at a limited ration). We found no evidence of local (thermal) adaptation, with warming affecting farm and forest populations similarly. Then, using mark–recapture methods to quantify impacts of warming on performance in situ, we found similar growth rates in fish from deforested and forested streams despite their distinct thermal environments. This suggests higher prey consumption at deforested sites to compensate for greater metabolic costs, which could strengthen fish–invertebrate interactions. Finally, we developed a bioenergetics model to estimate fish–invertebrate interaction strength and quantify temperature‐related and unrelated impacts of land use on this interaction. We found that although warming increased fish consumption, it apparently increased invertebrate production even more and thus had a net‐weakening effect on estimated interaction strength. Most importantly, variation in both fish and invertebrate density not directly related to temperature had a much stronger influence on estimated interaction strength than temperature effects on predator consumption and prey growth. We conclude that ectotherms can sometimes offset the metabolic costs of warming with a small increase in consumption that hardly effects food web interactions compared to non‐metabolic impacts of anthropogenic disturbances. Future research should assess whether this is a common feature of heavily impacted ecosystems facing multiple stressors. A plain language summary is available for this article.

In fisheries science, stock density is one of the fundamental factors that affect growth and condition. However, reproductive traits like fecundity and also the size and quality of offspring (via maternal effects) can be modified by stock density. In the short-living freshwater fish species vendace ( Coregonus albula ), the impact of stock density via maternal condition on quantity and quality of eggs and larvae was often proposed as a driver of population fluctuations, but such mechanisms have only rarely been evaluated. We systematically analysed growth, female condition and reproductive traits (fecundity, egg size, hatching rate and size of hatchlings) of vendace in relation to stock density across three lakes over two years. Across populations of contrasting density, length at first maturity, female condition and relative fecundity strongly reflected the difference in stock density, while size of eggs and larvae and hatching success at artificial breeding were not correlated with stock density. Our results indicate that density-dependent female spawner traits do not alter reproductive success, but vendace maintain egg size and hatching rate at the expense of egg quantity even at low per-capita resource availability. Accordingly, the reproductive potential of vendace stocks is primarily driven by spawning stock biomass and relative fecundity. We conclude that besides spawning stock biomass and relative fecundity, the additional inclusion of spawner condition into stock assessment has little potential to improve vendace fisheries management.

It is well documented that aquatic ecosystems may be subsidized by naturally derived terrestrial carbon sources. In contrast, the intentional or unintentional subsidy of animal populations by human-derived feed resources and their ecosystem effects are poorly studied. We added ground baits of the type, amount and temporal duration commonly applied by anglers targeting cyprinid fishes to a small lake, and studied behavior, diet composition and annual growth rate of the lake fish community in response to the bait addition. Based on recordings by a high-resolution ultrasonic telemetry array, common carp (Cyprinus carpio) as a model benthivore spend more time at the sites where ground baits were added, and they significantly reduced their home range relative to the period before bait addition. Furthermore, many omnivorous cyprinid fish species were regularly caught by angling at the feeding sites, indicating active ingestion of the artificially added food items. Stable isotope analyses of 13C and 15N showed that in particular larger specimens of common carp, bream (Abramis brama), white bream (Blicca bjoerkna), tench (Tinca tinca), roach (Rutilus rutilus) and rudd (Scardinius erythrophthalmus) included substantial amounts of angler baits into their diet. There was also a significant acceleration of growth in bream and white bream after bait addition, most likely attributable to the energetic benefits from bait addition. In contrast, there was no response in annual growth rate in top predators, suggesting they did not benefit from the subsidy. The amount of carbon introduced was very low (about 1%) relative to an estimate of in-lake C fixation by autotrophs. However, if the C added by bait was compared with the coarsely estimated secondary production of benthic macroinvertebrates in the lake, the C available to benthivorous and omnivorous fishes was comparable between natural resources and angler baits. We conclude that human-derived feed resources associated with recreational fishing may constitute a substantial subsidy to omnivorous fishes in lakes, in particular if the food is provided in form of particles, which are readily accessible and found at repeatable feeding places and over sufficiently long time periods during a year. The long-term consequences for the receiving ecosystems still have to be elucidated. peerReviewed

Human land use activities around estuaries can result in high levels of eutrophication. At Elkhorn Slough estuary, a highly eutrophic California estuary, we investigated the effects of impaired water quality on two stress-tolerant estuarine species, a common fish, the staghorn sculpin, Leptocottus armatus and a foundational invertebrate, the Olympia oyster, Ostrea lurida. We caged the two indicator species at six wetlands with different levels of water quality impairment, four of which had restricted tidal flow. We also recorded water quality parameters simultaneously at all sites using YSI sondes, and sampled nutrients and chlorophyll-a monthly, building on the National Estuarine Research Reserve System-wide Monitoring Program. We found that the monitored environmental variables predicted ecological responses by the indicator species. In particular, we found that the duration and severity of hypoxia were negatively correlated with fish survival and oyster growth. Further, our results corroborate previous studies that artificial tidal restriction leads to increased hypoxia stress. We conclude that large diurnal fluctuations in dissolved oxygen and extended nighttime hypoxia can have lethal and sub-lethal effects even on stress-tolerant organisms in the estuary. While laboratory experiments have often shown such effects, it is relatively rare to demonstrate negative effects of oxygen variation with in situ experiments, which provide stakeholders with concrete evidence for impaired water quality at local wetlands. Tidally restricted sites, which experience the largest fluctuations in dissolved oxygen and longest periods of hypoxia, harbor conditions harmful to vertebrates and invertebrates in the estuary. Reversing the anthropogenically induced low oxygen levels, by restoring more natural tidal exchange and by decreasing agricultural runoff, could improve the survival and growth of important estuarine organisms.

Animals typically display among-individual differences in behavior that are consistent over time (i.e., personality). These differences are often triggered by variable individual responses to environmental stress factors experienced during life, such as competition for resources and risk of predation. While the causes underlying animal personality are considered to be an issue of prime importance, it is still unknown whether personality emerges and develops over ontogeny if the main sources of behavioral differentiation are absent. Here, we tested whether personality emerged and was strengthened during the lifetime of Eastern mosquitofish (Gambusia holbrooki), once intraspecific competition and risk of predation were completely removed and genetic and maternal differences minimized. We found that individual differences in behavior were overall repeatable over ontogeny (i.e., personality was manifested). Personality was, however, not detectable in juvenile individuals but emerged during and after their sexual maturation. The emergence of personality was triggered by the decline in behavioral plasticity of individuals over ontogeny, while differences in behavior among individuals did not vary with age. Our results suggest that animal personality might be inevitable and emerge in fish under laboratory-controlled conditions even in absence of extrinsic factors that typically lead to behavioral differentiation. The decline of behavioral plasticity over lifetime might be a relevant mechanism for the development of personality in animals. Increasing evidence suggests that animals have personality, that is, individuals consistently differ in behavior among each other (e.g., bold and shy or social and non-social individuals). Personality differences among animals should be, by definition, consistent over time and often caused by environmental challenges experienced early in life. In this study, we observed that personality differences were not present at juvenile age in social fish but emerged later in their life, despite the fact that environmental challenges (i.e., predation risk and competition for space, food, and mates) were absent. Personality differences strengthened over lifetime, resulting from declines in individual behavioral plasticity. Our results suggest that the decline in behavioral plasticity with age may represent a relevant mechanism for behavioral differentiation in animals, in agreement with evidences from the human literature on age-related loss in behavioral plasticity.

Animals adjust their behaviour over time and contexts to cope with ecological and environmental variation. However, the presence of consistent between-individual differences in behaviour (i.e. personality) suggests that individual behaviour may be less flexible than previously thought. Here, we tested whether the size of the experimental tank and the ontogenetic stage of individuals affect estimates of average and consistent individual variation in activity and risk-taking behaviours in the eastern mosquitofish, Gambusia holbrooki, a fish model widely utilized for behavioural research. We measured risk taking and activity in juvenile, subadult and adult mosquitofish in a standard open-field test, in which the size of test tank varied linearly. We found strong evidence that spatial constraints alter mosquitofish behaviour. In particular, we observed that average activity increased with tank size, while the willingness of fish to take risks was independent of tank size. Moreover, juvenile fish exhibited, on average, lower risk-taking behaviours than older individuals. We highlight that the use of differently sized tanks may result in unequal variation in the average behaviour between juvenile and older fish, with escaping abilities of juvenile fish being underestimated in small environments. Most interestingly, we observed that variation in tank size triggered changes in the individual rank order for both risk taking and activity in juvenile fish, thus altering their personality estimates. In contrast, adult fish maintained consistent individual rank orders across all tank sizes. This study supports the hypothesis that behavioural repeatability increases with age, suggesting that personality estimates on adult animals may be less vulnerable to variation in laboratory contexts than those on juvenile ones.

Planktivorous and benthivorous fish have been documented to influence the density and size structure of their prey communities in lakes. We hypothesized that piscivorous fish modify their prey fish communities in the same way and sought to find evidence for such predation effects from a comparison across 356 lakes located in nine European ecoregions. We categorized individual fish as being piscivore, nonpiscivore, or prey of piscivores, depending on species and individual size. We calculated piscivore, nonpiscivore, and piscivore prey densities, respectively, and fit linear abundance size spectra (SS) on lake-specific piscivore, nonpiscivore, and piscivore-prey size distributions. Multiple linear regressions were calculated to quantify the effect of piscivore density and SS slopes on nonpiscivore and piscivore-prey densities and SS slopes by accounting for potentially confounding factors arising from lake morphometry, productivity, and local air temperature. Piscivore density correlated positively with piscivore-prey density but was uncorrelated with density of nonpiscivores. Across a subset of 76 lakes for which SS slopes of piscivores were statistically significant, SS slopes of piscivores were uncorrelated with SS slopes of either nonpiscivores or piscivore prey. However, densities of piscivores, nonpiscivores, or piscivore prey were a significant negative predictor of SS slopes of the respective groups. Our analyses suggest that direct predation effects by piscivorous fish on density and size structure of prey fish communities are weak in European lakes, likely caused by low predator–prey size ratios and the resulting size refuges for prey fish. In contrast, competition may substantially contribute to between-lake variability in fish density and size.

This study tested for links among behaviour, state and life-history variables as predicted by the pace-of-life hypothesis in adult pike Esox lucius. First, a standardized open-field behavioural assay was developed to assess individual behaviour of wild-captured adult E. lucius. Behaviour within the standardized assay predicted swimming behaviour in the lake, providing an ecological validation of the assay. There was no relationship between standardized behaviour and any of the life-history and state variables, including metabolism, body condition, juvenile growth rate and adult growth rate in contrast to predictions from the pace-of-life hypothesis. This study demonstrates that it is possible to assess ecologically relevant behavioural variation in a large-bodied top predator using a standard open-field assay, but it is noteworthy that this standardized behaviour is not systematically related to standard metabolism or growth.

Compartmentalized microfluidic devices with immobilized catalysts are a valuable tool for overcoming the incompatibility challenge in (bio) catalytic cascade reactions and high-throughput screening of multiple reaction parameters. To achieve flow control in microfluidics, stimuli-responsive hydrogel microvalves were previously introduced. However, an application of this valve concept for the control of multistep reactions was not yet shown. To fill this gap, we show the integration of thermoresponsive poly(N-isopropylacrylamide) (PNiPAAm) microvalves (diameter: 500 and 600 &micro, m) into PDMS-on-glass microfluidic devices for the control of parallelized enzyme-catalyzed cascade reactions. As a proof-of-principle, the biocatalysts glucose oxidase (GOx), horseradish peroxidase (HRP) and myoglobin (Myo) were immobilized in photopatterned hydrogel dot arrays (diameter of the dots: 350 &micro, m, amount of enzymes: 0.13&ndash, 2.3 &micro, g) within three compartments of the device. Switching of the microvalves was achieved within 4 to 6 s and thereby the fluid pathway of the enzyme substrate solution (5 mmol/L) in the device was determined. Consequently, either the enzyme cascade reaction GOx-HRP or GOx-Myo was performed and continuously quantified by ultraviolet-visible (UV-Vis) spectroscopy. The functionality of the microvalves was shown in four hourly switching cycles and visualized by the path-dependent substrate conversion.

We propose an improved micro reactor design for a scalable microfluidic device, in which enzymes are immobilized in a hydrogel matrix. Furthermore, fluid flow is controlled by means of hydrogel-based micro-valves. In this work, computational flow simulations will be compared to experimental results to highlight new design ideas and to improve wetting and concentration distribution through the entire chamber volume, even for high aspect ratios. Additionally, modelling concepts will be introduced to efficiently describe multi-domain problems like enzyme reactions. With the help of a computer-aided design process which is capable to simulate hydrogel-based microfluidic systems it is possible to better understand, predict and visualize the behavior of micro-reactors and support the development of highly integrated hydrogel-based microfluidic circuits.

[Departement_IRSTEA]Eaux [TR1_IRSTEA]QUASARE [ADD1_IRSTEA]Systèmes aquatiques soumis à des pressions multiples; International audience; Body size, coupled with abundance and taxonomy, may help to understand the mechanisms shaping community structure. Since the body size of fish is closely related to their trophic niche, size diversity (based on individual body size) of fish communities may capture intraspecific variations in fish trophic niches that are not detected by species diversity. Thus, the relationship between size diversity and species diversity may help to integrate variation at both intraspecific and interspecific levels. We studied the relationship between species diversity and size diversity as a measure of the degree of overlap in size among species and thereby the potential overlap in niches in a community. We hypothesized that the relationship between size diversity and species would be different across the European continent due to different levels of size overlap in fish communities. The data were derived from samplings of fish communities using standardised benthic gill nets in 363 lakes. At the continental scale, size diversity increased with species diversity; at the ecoregion scale, the slope of the relation changed across the continent, with the greatest mismatch occurring in northern Europe where communities comprised only one or a few species, but each of which exhibited a great range in size. There was an increase in slope towards the south with significant relations for four out of six ecoregions. The steeper size diversity-species diversity slope at lower latitudes is attributable to a lower overlap in fish size and thus likely to finer niche separation. Our results also suggest that size diversity is not a strong surrogate for species diversity in European lake fish communities. Thus, particularly in fish communities composed of few species, measuring size diversity may help to detect potential functional variation which may be neglected by measuring species diversity alone.

Invasion of non-native species is considered a major threat to global biodiversity. Here we present a comprehensive overview of the occurrence, richness and biomass contribution of non-native fish species in 1943 standing water bodies from 14 countries of the Western Palearctic, based on standardised fish catches by multi-mesh gillnetting. We expected strong geographical gradients to emerge in the occurrence of non-natives. We further hypothesised that the contribution by non-natives to the local fish community biomass was correlated with local richness and the trophic level of native and non-native species. Non-native fish species occurred in 304 of 1943 water bodies (16%). If the average number of occupied water bodies per country was weighted by number of water bodies per country, the grand mean occurrence of non-natives in Western Palearctic water bodies was 10%. Exotic (non-native to the Palearctic) and translocated (non-native only to parts of the Palearctic) species were found in 164 (8.4%) or 235 (12.1%) of the water bodies, respectively. The occurrence and local richness of non-native fish species increased with temperature, precipitation and lake area and were substantially higher in reservoirs than in natural lakes. High local biomass contributions of non-native species were strongly correlated with low richness of native species and high richness of non-native species, whereas the trophic level of the fish species had only a weak effect. Single non-native species rarely dominated community biomass, but high biomass contributions and thus strong community and ecosystem impacts can be expected if several non-native species accumulate in a water body. Fil: Trochine, Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; Argentina. Universidad Nacional del Comahue. Centro Regional Universitario Bariloche. Laboratorio de Limnología; Argentina Fil: Brucet, Sandra. University of Vic; España. Catalan Institution for Research and Advanced Studies; España. Institució Catalana de Recerca i Estudis Avancats; España Fil: Argillier, Christine. Aix En Provence; Francia Fil: Arranz, Ignasi. University of Vic; España Fil: Beklioglu, Meryem. Middle East Technical University; Turquía Fil: Benejam, Lluís. University of Vic; España Fil: Ferreira, Teresa. Universidade de Lisboa; Portugal Fil: Hesthagen, Trygve. Norwegian Institute for Nature Reasearch; Noruega Fil: Holmgren, Kerstin. Swedish University of Agricultural Sciences; Suecia Fil: Jeppesen, Erik. Sino-danish Centre For Education And Research; China. University Aarhus; Dinamarca Fil: Kelly, Fiona. Inland Fisheries Ireland; Irlanda Fil: Krause, Teet. Estonian University of Life Sciences; Estonia Fil: Rask, Martti. Natural Resources Institute Finland; Finlandia Fil: Volta, Pietro. Consiglio Nazionale delle Ricerche; Italia Fil: Winfield, Ian J.. Lancaster Environment Centre; Reino Unido Fil: Mehner, Thomas. Leibniz - Institute of Freshwater Ecology and Inland Fisheries; Alemania

Summary Phage shock protein A (PspA) belongs to the highy conserved PspA/IM30 family and is a key component of the stress inducible Psp system in Escherichia coli. One of its central roles is the regulatory interac- tion with the transcriptional activator of this system, the σ 54 enhancer-binding protein PspF, a member of the AAA+ protein family. The PspA/F regulatory system has been intensively studied and serves as a paradigm for AAA+ enzyme regulation by trans-acting factors. However, the molecular mechanism of how exactly PspA controls the activity of PspF and hence σ 54 -dependent expression of the psp genes is still unclear. To approach this question, we identified the minimal PspF-interacting domain of PspA, solved its structure, determined its affinity to PspF and the dis- sociation kinetics, identified residues that are poten- tially important for PspF regulation and analyzed effects of their mutation on PspF in vivo and in vitro. Our data indicate that several characteristics of AAA+ regulation in the PspA·F complex resemble those of the AAA+ unfoldase ClpB, with both proteins being regulated by a structurally highly conserved coiled- coil domain. The convergent evolution of both regu- latory domains points to a general mechanism to control AAA+ activity for divergent physiologic tasks via coiled-coil domains.

Turbidity, caused by suspended particles in the water column, induces light scattering and shifts in the wavelengths of light. These changes may impair the ability of fish to use physical cues and hence may modify social interactions. We experimentally investigated the social interactions of guppies, Poecilia reticulata, in clear and turbid water. Fish were significantly less active, formed smaller shoals and were found to be more often alone in turbid than in clear water. A Markov chain analysis revealed significant differences in the social dynamics when comparing clear and turbid water conditions. The probability of leaving a particular nearest neighbour and the probability of choosing some neighbour after swimming around alone differed between the treatments. Our results indicate that turbidity has a number of different effects on the social interactions of the guppy, and we discuss their potential costs and benefits and wider implications.

Background Limited effectiveness of therapeutic agents targeting epidermal growth factor receptor (EGFR) in clinical trials using unselected ovarian cancer patients has prompted efforts to more effectively stratify patients who might best benefit from these therapies. A series of studies that have evaluated immunohistochemical (IHC) staining of EGFR in ovarian cancer biopsies has produced unclear results as to the utility of this measure as a prognostic biomarker. Here, we used one of the largest, single institution cohorts to date to determine possible associations of EGFR expression with patient outcome. Methods We performed IHC staining of EGFR in tissue microarrays including nearly 500 patient tumor samples. Staining was classified by subcellular localization (membranous, cytoplasmic) or by automated image analysis algorithms. We also performed a literature review to place these results in the context of previous studies. Results No significant associations were found between EGFR subcellular localization or expression and histology, stage, grade, or outcome. These results were broadly consistent with the consensus of the reviewed literature. Conclusions These results suggest that IHC staining for EGFR may not be a useful prognostic biomarker for ovarian cancer patients. Future studies should pursue other staining methods or analysis in combination with other pathway mediators.

Body size of consumer species is a fundamental trait that influences the trophic ecology of individuals and their contribution to the functioning of freshwater ecosystems. However, the relationship between body size and trophic ecology can be highly variable both within and between closely-related and similarly-sized species. In this study we compared the intra- and interspecific relationship between body size and trophic position for North American Yellow Perch Perca flavescens and European Perch Perca fluviatilis, which share similarities in morphology, life history traits and trophic requirements. We used stable isotope ratios (δ15N and δ13C) to characterize differences in size-dependency of trophic position and to trace consumer foraging history of Yellow Perch in lakes in the Northwestern United States and European Perch in lakes in Germany. The trophic position and stable isotope ratios of Yellow Perch and European Perch steadily increased with total body length, but European Perch were consistently feeding at higher trophic positions than Yellow Perch at a given length. European Perch occupied considerably higher trophic positions (mean trophic position = 3.9) than Yellow Perch (mean trophic position = 2.8). Large European Perch were increasingly piscivorous, whereas large Yellow Perch were more opportunistic and omnivorous predators of invertebrate prey. Overall, the trophic position among individual Yellow Perch varied more strongly than in European Perch. We conclude that both species similarly increase in trophic position with size, but the specific size-dependency of both trophic position and resource use varies with taxonomy and local ecological conditions. Thus, body size as a sole measure of trophic position should be considered cautiously when generalizing across populations and species.

We compared the potential of different lake typologies to discriminate fish communities in least disturbed sites. The typologies tested were based on morphometric and geographical descriptors. The best discrimination was achieved by distinguishing three lake types according to depth and mixes regime: polymictic lakes, stratified lakes with less than 30 m of maximum depth and deep, stratified lakes with maximum depths above 30 m. We conclude that the proposed typology is appropriate for a system to assess the ecological status of German lakes with the fish fauna according to the Water Framework Directive and might well be transferable to other European assessment systems. The fish communities in all lake types were similar and dominated by few fish species. Perch and roach were the most abundant ones, followed by ruffe, bream, rudd and pike. The fish communities in least disturbed sites might be used as reference conditions in future fish-based assessment systems.

Many ecosystems are linked to their adjacent ecosystems by movements of organisms. For instance, aquatic and terrestrial ecosystems are linked via emerging aquatic insects that serve as prey for terrestrial consumers. However, the role of these organisms in returning recycled carbon to the ecosystem from which it originated is not well known. This is due to the fact that values of carbon isotope signatures from terrestrial leaves and aquatic resources are usually similar and hence results of isotope mixing models need to be considered with caution. We overcame this problem by adding isotopically distinct terrestrial particulate organic carbon (tPOC) as a tracer to the experimental sides of two lakes that were divided in two equal halves with plastic curtains. We focused on aquatic insect larvae (Chironomidae) that fed on maize Zea mays leaves experimentally added to the lakes, and subsequently became prey for terrestrial predators (spiders) after emergence. The carbon isotope values of Chironomidae and spiders were significantly elevated in the lake treatment sides as compared to reference sides, whereas the values of all autochthonous resources were not affected by maize additions. Estimates from stable isotope mixing models indicated a low but demonstrable contribution of maize leaves to the diet of Chironomidae. Overlap between the isotope values of alder leaves, the major natural tPOC source, and autochthonous resources prevented a reliable quantification of allochthony of Chironomidae. However, we qualitatively demonstrated the flow of terrestrial particulate organic carbon to lakes, as leaf fall, and back to terrestrial surroundings via emerging insects. This ‘boomerang’ carbon flux between land and lakes blurs the distinction between autochthonous and allochthonous carbon sources.

// Christine Mehner 1 , Alexandra Hockla 1 , Erin Miller 1 , Sophia Ran 2 , Derek C. Radisky 1,* and Evette S. Radisky 1,* 1 Department of Cancer Biology, Mayo Clinic, Jacksonville, FL 2 Department of Medical Microbiology, Immunology and Cell Biology, Southern Illinois University School of Medicine, Springfield, IL Correspondence: Evette S. Radisky, email: // Derek C. Radisky, email: // Keywords : triple negative breast cancer, basal-like breast cancer, matrix metalloproteinases, invasion, metastasis, mouse orthotopic tumor models, bioluminescence imaging Received : March 3, 2014 Accepted : April 30, 2014 Published : May 1, 2014 Abstract Matrix metalloproteinases (MMPs) have been implicated in diverse roles in breast cancer development and progression. While many of the different MMPs expressed in breast cancer are produced by stromal cells MMP-9 is produced mainly by the tumor cells themselves. To date, the functional role of tumor cell-produced MMP-9 has remained unclear. Here, we show that human breast cancer cell-produced MMP-9 is specifically required for invasion in cell culture and for pulmonary metastasis in a mouse orthotopic model of basal-like breast cancer. We also find that tumor cell-produced MMP-9 promotes tumor vascularization with only modest impact on primary tumor growth, and that silencing of MMP-9 expression in tumor cells leads to an altered transcriptional program consistent with reversion to a less malignant phenotype. MMP-9 is most highly expressed in human basal-like and triple negative tumors, where our data suggest that it contributes to metastatic progression. Our results suggest that MMP9 may offer a target for anti-metastatic therapies for basal-like triple negative breast cancers, a poor prognosis subtype with few available molecularly targeted therapeutic options.

Background: Breast cancer often metastasizes into bone and leads to osteolytic lesions. The underlying mechanisms, however, are complex and not fully understood. Syndecan-1 is a proteoglycan that has various functions relevant for tumor progression including cell–cell communication and cell–matrix interactions. Moreover, its two glycosaminoglycan-binding sites suggest that it may interfere with glycoproteins such as osteoprotegerin, a potent inhibitor of osteoclastogenesis. Thus, we hypothesize that tumor-derived syndecan-1 alters osteoclast biology by modulating osteoprotegerin. Methods: Syndecan-1 expression was down-regulated via siRNA and the cell fate of the breast cancer cell lines MCF-7, T-47D, and MDA-MB-231 was investigated. Furthermore, we determined the regulation of syndecan-1 by dexamethasone, a commonly used antiemetic in breast cancer therapy. Additionally, we analyzed the genesis and activity of osteoclasts in indirect co-culture experiments using supernatants from MCF-7 cells with deficient and sufficient levels of syndecan-1. Results: Dexamethasone time- and dose-dependently increased syndecan-1 expression up to 4-fold but did not alter cell behavior. Syndecan-1 up-regulation did not affect the survival or migration of breast cancer cells. Depletion of syndecan-1 using siRNA led to decreased vitality of progesterone receptor-positive cell lines. In MCF-7 cells osteoprotegerin production was up-regulated 2.5-fold after syndecan-1 knock-down. The culture of osteoclast precursors with the supernatant of MCF-7 cells with reduced syndecan-1 levels suppressed osteoclast formation and activity by 21% and 23%, respectively. Adding neutralizing antibodies to osteoprotegerin to the breast cancer supernatants reversed osteoclastogenesis. Conclusion: Thus, we identified tumor-derived syndecan-1 as a novel positive regulator of osteoclastogenesis and new player in the tumor-bone dialog.

An important challenge in community ecology is identifying the functional characteristics capable of predicting the nature and strength of predator effects on food webs. We developed an individual-based model, based on a shallow lake model system, to evaluate the total, consumptive, and non-consumptive indirect effect that predators have on basal resources when the predators differ in their foraging types (active adaptive foraging or sedentary foraging). Overall, both predator types caused similar total indirect effects on lower trophic levels. However, the nature net effects of predators diverged between predator foraging types. Active predators caused larger non-consumptive effects, relative to the total indirect effect, irrespective of predation pressure levels. On the other hand, sedentary predators caused larger nonconsumptive effects for lower predation pressure levels, but consumptive effects became more important as predation pressure increased. Our simulations showed that the reliance on a particular mechanism driving consumer‐resource interactions is altered by predator foraging behavior and highlight the importance of both prey and predator foraging behaviors to predict the causes and consequences of cascading effects observed in food webs.

Basal subtype breast cancers have a particularly poor prognosis, with high invasiveness and resistance to most targeted therapies. TGFβ and MYC drive central features of basal breast cancer: TGFβ is an autocrine and paracrine signaling factor that drives cell invasion and metastasis, and MYC is a central regulator of cellular proliferation that is upregulated in many cancer types. We show here that genetic or pharmacologic inhibition of MYC in MCF10A basal breast cells results in increased sensitivity to TGFβ-stimulated invasion and metastasis and also show that this signaling loop is dependent on activation of SRC. Analysis of human breast cancer datasets and additional experiments with breast cancer cell lines further suggest the relevance of this signaling loop in basal, but not luminal, breast cancers. Our results imply precaution should be taken when utilizing therapeutic inhibitors of MYC with basal breast cancer patients as this could lead to increased metastasis; however, simultaneous pharmacologic inhibition of SRC and MYC for these patients could facilitate the antiproliferative effects of MYC inhibition while blocking the consequent promotion of metastasis. Cancer Res; 76(12); 3520–30. ©2016 AACR.

Tat systems transport folded proteins across energized membranes of bacteria, archaea, and plant plastids. In Escherichia coli, TatBC complexes recognize the transported proteins, and TatA complexes are recruited to facilitate transport. We achieved an abstraction of TatA from membranes without use of detergents and observed a co-purification of PspA, a membrane-stress response protein. The N-terminal transmembrane domain of TatA was required for the interaction. Electron microscopy displayed TatA complexes in direct contact with PspA. PspB and PspC were important for the TatA-PspA contact. The activator protein PspF was not involved in the PspA-TatA interaction, demonstrating that basal levels of PspA already interact with TatA. Elevated TatA levels caused membrane stress that induced a strictly PspBC- and PspF-dependent up-regulation of PspA. TatA complexes were found to destabilize membranes under these conditions. At native TatA levels, PspA deficiency clearly affected anaerobic TMAO respiratory growth, suggesting that energetic costs for transport of large Tat substrates such as TMAO reductase can become growth limiting in the absence of PspA. The physiological role of PspA recruitment to TatA may therefore be the control of membrane stress at active translocons.

Summary 1. Diel vertical migrations (DVM) are typical for many cold-water fish species such as Pacific salmons (Oncorhynchus spp.) and coregonids (Coregonus spp.) inhabiting deep lakes. A comprehensive recent overview of DVM in freshwater fish has not been available, however. 2. The main proximate trigger of DVM in freshwater fish is the diel change in light intensity, with declining illumination at dusk triggering the ascent and the increase at dawn triggering the descent. Additional proximate cues are hydrostatic pressure and water temperature, which may guide fish into particular water layers at night. 3. Ultimate causes of DVM encompass bioenergetics efficiency, feeding opportunities and predator avoidance. None of these factors alone can explain the DVM in all cases. Multi-factorial hypotheses, such as the ‘antipredation window’ combined with the thermal niche hypothesis, are more likely to explain DVM. It is suggested that planktivorous fish move within a layer sufficiently well illuminated to capture zooplankton, but too dark for predators to feed upon the migrating fish. In complete darkness, fish seek layers with a temperature that optimises bioenergetics efficiency. The strength of each factor may differ from lake to lake, and hence system-specific individual analyses are needed. 4. Mechanistic details that are still poorly explored are the costs of buoyancy regulation and migration, the critical light thresholds for feeding of planktivorous and piscivorous fish, and predator assessment by (and size-dependent predation risk of) the prey fish. 5. A comprehensive understanding of the adaptive value of DVM can be attained only if the behaviour of individual fish within migrating populations is explicitly taken into account. Size, condition and reproductive value differ between individuals, suggesting that migrating populations should split into migrants and non-migrants for whom the balance between mortality risk and growth rate can differ. There is increasing evidence for this type of partial DVM within populations. 6. Whereas patterns of DVM are well documented, the evolution of DVM is still only poorly understood. Because experimental approaches at realistic natural scales remain difficult, a combination of comprehensive data sets with modelling is likely to resolve the relative importance of different proximate and ultimate causes behind DVM in fish.

Diel vertical migration (DVM) in fish has been studied extensively on the population level, but the individual variability of migration behavior has been widely neglected. However, the individual size of fish modifies the ratio between mortality risk and foraging gain. Therefore, if the rhythmic habitat shifts are considered being adaptive, the average time spent in the nighttime shallow habitat should differ between small and large fish, suggesting that the individual migration trajectories might be size dependent. We studied size-related differences in the vertical migration speed and the onset of DVM in coregonid fishes (Coregonus spp.) in a deep lake. Stationary vertical hydroacoustics was conducted once a month over a 7-month period to track individual fish during the migration periods at dusk and dawn. Quantile regressions revealed that vertical migration speed was higher for smaller than for larger fish. The comparison of detection times of fish migrating upward after sunset revealed an earlier detection of small fish compared with the medium and large fish, thus suggesting that smaller fish ascended earlier and faster at dusk. The reverse trend at dawn was less clearly expressed. Bioenergetics calculations suggest that small fish may profit from a faster and earlier ascent by increasing their daily ration by about 8% relative to fish that would migrate together with the main population. Therefore, plasticity of migration speeds may result in individual competitive advantages and heterogeneous mortality-feeding trade-offs within populations. This variability in individual migration trajectories has to be considered in future studies addressing ultimate causes of DVM. [Behav Ecol]

Metabolism constitutes a fundamental property of all organisms. Metabolic rate is commonly described to scale as a power function of body size and exponentially with temperature, thereby treating the effects of body size and temperature independently. Mounting evidence shows that the scaling of metabolic rate with body mass itself depends on temperature. Acrossspecies analyses in fishes suggest that the mass-scaling exponent decreases with increasing temperature. However, whether this relationship holds at the within-species level has rarely been tested. Here, we re-analyse data on the metabolic rates of four freshwater fish species, two coregonids and two cyprinids, that cover wide ranges of body masses and their naturally experienced temperatures. We show that the standard metabolic rate of the coregonids is best fit when accounting for a linear temperature dependence of the scaling of metabolic rate with body mass, whereas a constant mass-scaling exponent is supported in case of the cyprinids. Our study shows that phenotypic responses to temperature can result in temperaturedependent scaling relationships at the species level and that these responses differ between taxa. Together with previous findings, these results indicate that evolutionarily adaptive and phenotypically plastic responses to temperature affect the scaling of metabolic rate with body mass in fishes. Metabolism constitutes a fundamental property of all animals and plants as it supplies an organism with energy and materials from its environment. The most important factors influencing the metabolic rate are body mass and temperature. In poikilotherms whose body temperature varies along with that of the environment, metabolism links the ambient temperature to intrinsic physiological processes. This interaction between environment and individual metabolism affects ecological and evolutionary processes at all levels of organization. It has been argued that in aerobic eukaryotes body mass and temperature, through the effects on metabolism, explain most of the varia

The aim of our study was to elucidate the role of wavelength and irradiance in blue light retinal damage. We investigated the impact of blue light emitted from light-emitting diode (LED) modules with peaks at either 411nm (half bandwidth 17nm) or 470nm (half bandwidth 25nm) at defined irradiances of 0.6, 1.5 and 4.5W/m(2) for 411nm and 4.5W/m(2) for 470nm on retinal neuronal (R28) cells in vitro. We observed a reduction in metabolic activity and transmembrane potential of mitochondria when cells were irradiated at 411nm at higher irradiances. Furthermore, production of mitochondrial superoxide radicals increased significantly when cells were irradiated with 411nm light at 4.5W/m(2) . In addition, such irradiation caused an activation of the antioxidative glutathion system. Using vital staining, flow cytometry and western blotting, we were able to show that apoptosis only took place when cells were exposed to 411nm blue light at higher irradiances; necrosis was not observed. Enhanced caspase-3 cleavage product levels confirmed that this effect was dependent on light irradiance. Significant alterations of the above-mentioned parameters were not observed when cells were irradiated with 471nm light despite a high irradiance of 4.5W/m(2) , indicating that the cytotoxic effect of blue light is highly dependent on wavelength. The observed phenomena in R28 cells at 411nm (4.5W/m(2) ) point to an apoptosis pathway elicited by direct mitochondrial damage and increased oxidative stress. Thus, light of 411nm should act via impairment of mitochondrial function by compromising the metabolic situation of these retinal neuronal cells.

Recent studies on diel vertical migration (DVM) of two coregonid species ( Coregonus spp.) in Lake Stechlin (Germany) have suggested that the nocturnal distribution of fish is linked with metabolic benefits. We used a spatially explicit bioenergetics-based foraging model to test whether energetic constraints contribute to explain DVM of both species. The newly parameterized model was compared with independent data from Lake Stechlin that covered a 9 month period. Predicted growth rates matched observed growth rates of each fish species reasonably well. The simulation of different migration scenarios showed that even slight changes in night-time depths modified growth rates, primarily owing to temperature-dependent respiration. Fish that performed DVM grew faster than nonmigratory fish that occupied deep hypolimnetic water over a diel cycle. However, the most energy-efficient strategy simulated had fish remaining within the metalimnion. Here, energetic benefits (foraging) clearly outweighed higher energetic costs (respiration) in the warmer waters. Although DVM can be energetically beneficial, it is not the most efficient strategy performed by coregonids in Lake Stechlin. We suggest that multiple factors, rather than bioenergetics efficiency alone, are the evolutionary basis for DVM of many freshwater fish species.

Summary 1. Diel vertical migrations (DVM) are frequently observed in pelagic fish and zooplankton populations. In addition to predator avoidance and foraging opportunity, bioenergetics optimisation has been hypothesised to cause the selective advantage of migrating fish. However, experimental confirmation of growth advantages of fish held at naturally observed cyclic temperatures, and food densities are surprisingly rare. 2. We compared growth rates, growth efficiencies and energy budgets of vendace (Coregonus albula, Salmoniformes) fed daily rations of 10% body mass and held at low (4.5 °C), high (8 °C) and cyclic (switch between 4.5 and 8 °C) temperatures over a period of 6 weeks. Biochemical body composition was evaluated by bioimpedance analyses and direct determination of water, lipid and protein content in a subset of fish. 3. Growth rate and food conversion efficiency were similar in fish held at high and cyclic temperatures, but higher than those found at low temperatures. Body condition of fish at cyclic temperatures was maximised, but these fish also had the highest water content. The proportion of lipids and proteins was slightly depleted in fish held at cyclic and low temperatures relative to the high temperature treatment. Accordingly, growth and biochemical composition of fish responded specifically to cyclic temperatures and were not simply intermediate between those found at low and high temperatures. 4. We could not confirm a bioenergetics advantage of fish switching between high and low temperatures. However, there were no substantial extra metabolic costs through exposure to cyclic temperatures, and hence, fish performing DVM may benefit from predation avoidance without compromising their metabolic balance and hence growth rates. These results suggest that the evolution of DVM is a multi-faceted process with no single ultimate explanation.

Summary 1. Field studies on diel vertical migration (DVM) usually report uniform behaviour with population-wide ascents and descents during crepuscular periods. This contrasts partial seasonal migrations of many animal populations, where individuals choose either the resident or the migrant strategy depending on population density, feeding opportunity and predation risk in the resident and migrant habitats. 2. We tested whether DVM of freshwater zooplanktivorous fish (Coregonus spp.) resembles partial migrations. Twenty-eight hydroacoustic surveys were performed in the deep Lake Stechlin (Germany) between 2000 and 2010, with samplings encompassing all months between March and December. Zooplankton samples were simultaneously taken in epilimnetic and hypolimnetic layers. Fish obtained from depth-stratified samplings by a midwater trawl were used to test for individual differences between residents and migrants. 3. We show for the first time that DVMs of freshwater fish resemble patterns of partial migrations often found in seasonal environments. Across all samplings, 7–33% of fish did not ascend at dusk, but exhibited the resident strategy. The proportion or residents increased at low zooplankton feeding rates in the daytime habitat and during months when the temperature difference between daytime and night-time habitats was minor. 4. Slightly larger size and higher caloric density of migrants over residents in one of the coexisting Coregonus species suggested that individual differences contributed to the migration strategy performed. However, these results were based on one sample only, and extrapolation to the entire data set is not possible. 5. Our results are indirect evidence that the balance between migrants and residents may primarily depend on the trade-off between feeding gains and metabolic and predation costs of migration. However, the results also suggest that the global fitness consequences for the resident and migrant strategies may not be identical, rendering the importance of individual traits in the ‘decision to migrate’ likely.

This special issue of Evolutionary Ecology provides ten papers that have been presented at a conference on Evolutionary Ecology of Fishes in 2009. In addition to briefly summarizing the main content of the papers which is related to adaptive radiations, processes of ecological divergence, and fisheries-induced evolution, we review and synthesize in short the recent advance in studies on evolutionary ecology of fishes. We conclude that fishes are excellent model systems to study evolutionary ecology of animals, and suggest three promising new research avenues; (1) the contribution of behavioural processes to evolution, in particular the consideration of animal personalities and predator–prey interactions, (2) metabolic physiology and parasite-host interactions as new niche dimensions to be considered for adaptive diversification, and (3) the opportunities for mechanistic understanding of adaptation and speciation emerging from new genetic tools.

Bernd Wefelmeyer, born in 1940, is a composer, arranger and conductor. He studied composition at the Hanns Eisler College of Music in Berlin with Ruth Zechlin, Wolfram Heicking and Rudolf Wagner-Regeny and worked as a jazz pianist (with Wolf Biermann, Manfred Krug and others). From 1966 to 1972, he was a sound engineer for Berlin Radio. After being dismissed from his radio position, he worked as a freelance composer, conductor and arranger—mainly for stage, film and chorus music, in addition to works for other media. Starting in 1991, Wefelmeyer worked as a professor, first at the Hanns Eisler College of Music and later also at the College of Film in Potsdam-Babelsberg.

Mehner T, Busch S, Helland IP, Emmrich M, Freyhof J. Temperature-related nocturnal vertical segregation of coexisting coregonids. Ecology of Freshwater Fish 2010: 19: 408–419. © 2010 John Wiley & Sons A/S Abstract – Habitat choice of fish may be influenced by many different ecological factors, e.g., predation risk, feeding opportunity, or temperature and oxygen availability. However, because most of the fish prey and their predators rely on vision for feeding, the predator avoidance and feeding opportunity hypotheses may fail to predict distribution of fish at complete darkness. Here, we accumulated patterns of nocturnal vertical distribution of two coexisting coregonid populations in Lake Stechlin from 13 samplings over 4 years, conducted by hydroacoustics and simultaneous midwater trawling. We calculated population depths, dispersion, illumination strengths and vertical temperature gradients for all sampling dates. Illumination strengths at fish population depths were far below the critical levels for feeding by vision, suggesting that predator avoidance or feeding opportunity did not trigger the depth distribution at night. In contrast, nocturnal population depths and dispersion of vendace Coregonus albula were significantly associated with the seasonally changing vertical temperature gradient in Lake Stechlin, whereas night-time distribution of the coexisting Fontane cisco Coregonus fontanae was almost unaffected by temperature. Vendace occurred just below the thermocline in isothermal water layers of about 6.5–9 °C during stratification of Lake Stechlin, whereas Fontane cisco preferred 4–6 °C cold layers. These experienced temperatures roughly correspond to species-specific optimum metabolic temperatures determined in earlier experiments. We assume, therefore, that the temperature-related vertical segregation during non-feeding hours at darkness is linked with metabolic benefits, thus suggesting that bioenergetics efficiency contributes to ultimate causes of diel vertical migrations at least in vendace.

Patients diagnosed with ovarian cancer face an average 5 year survival rate of 46%; low survival is driven by early and ongoing intraperitoneal dissemination and metastasis of the tumor. Among the different histotypes of ovarian cancer, ovarian clear cell carcinoma (OCCC) has a particularly poor prognosis when diagnosed in late stage, as these tumors tend to be chemoresistant, leaving no effective therapeutic options. Intraperitoneal metastasis requires cells to become anoikis resistant, where apoptosis normally induced by loss of attachment is suppressed. We seek to better understand the mechanisms involved in anoikis resistance in OCCC, which in the future could lead to new therapeutic strategies for these patients. We have found that Serine Protease Inhibitor Kazal type 1 (SPINK1), an endogenous inhibitor of trypsin like serine proteases, is also an important regulator of anoikis resistance in some ovarian cancers. In this study, we aimed to dissect the role of SPINK1 specifically in OCCC, defining its contribution to anoikis resistance using cultured OCCC cell lines and elucidating its mechanism of action. We compared anoikis resistance of cells with endogenous SPINK1 expression versus cells in which SPINK1 was silenced using lentiviral shRNA constructs, when culturing cells on ultra-low attachment plates to mimic cell detachment from the extracellular matrix. We found that knockdown of SPINK1 reduced survival and stimulated apoptotic pathways in OCCC cells grown under detached conditions, implicating SPINK1 in anoikis resistance of OCCC cells. As a secreted protease inhibitor, SPINK1 may be expected to confer anoikis resistance by inhibiting an extracellular serine protease involved in triggering anoikis. Because many proteases represent potential targets of SPINK1, we have designed a strategy using Activity Based Protein Profiling (ABPP) to discover the targets of SPINK1 from among this large pool of candidates. In pilot studies, we have successfully used a novel activity-based probe to covalently label the active sites of serine proteases secreted from OCCC cells. In ongoing efforts, we are optimizing methods to identify labeled proteases with high sensitivity using tandem mass spectrometry. We will then identify those proteases regulated by SPINK1 through comparison of labeled proteomes from OCCC cells with and without SPINK1 treatment,, and candidate proteases will be further analyzed for their role in triggering anoikis. The identification of SPINK1 regulated proteases that are responsible for mediating anoikis in our models will give insight into important mechanisms whereby tumor cells acquire resistance to anoikis in OCCC. In-depth understanding of anoikis resistance, a critical component of ovarian cancer progression and metastasis, may lead to novel biomarkers of disease progression as well as important therapeutic targets. Citation Format: Christine Mehner, Mathew A. Coban, Alexandra Hockla, Derek C. Radisky, Evette S. Radisky. Serine protease inhibitor Kazal type 1 (SPINK1) drives anoikis resistance in ovarian clear cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 442.

Navigation-induced physical forces have been suggested to modify the structure of riverine fish assemblages by impeding especially the recruitment of littoral bound species. To investigate the effect of vessel frequency on fish, we compared the composition and seasonal succession of young-of-the-year (YOY) fish assemblages in three similarly degraded river reaches differing in average vessel passages (2, 6 and 41 per day). Fish were caught by electrofishing biweekly between May and September. Multivariate tests were used to analyse differences between YOY-fish assemblages and hurdle regression models applied to determine abiotic factors predicting fish occurrence and abundance. Roach (Rutilus rutilus) and perch (Perca fluviatilis) densities were compared. Roach larvae remain in the littoral zone while perch larvae shift to the pelagic zone immediately after hatch. YOY-fish assemblage structure substantially changed along the traffic intensity gradient. In the high traffic intensity reach, species number and total fish density were markedly reduced compared to the other reaches. Roach densities were lowest in the high traffic intensity reach whereas perch densities did not decline along the gradient. Hurdle regressions confirmed a stronger effect of commercial navigation traffic intensity on roach than on perch. The total zooplankton biomass was highest in the high traffic intensity reach. Our results provide empirical evidence that intensive commercial navigation impoverishes fish assemblages in width-restricted waterways. They underlined that in particular those species that have their first nursery habitats in shoreline areas were more affected by intensive commercial navigation than species whose larvae live predominantly pelagic. The results indicate that the negative effect of intensive navigation on riverine fish results primarily from the navigation-induced hydraulic disturbances along the banks. Therefore, mitigation of navigation-induced hydraulic forces is required to prevent degradation of fish communities in waterways. Copyright © 2010 John Wiley & Sons, Ltd.