Assessing the Influence of Salmon Farming through Total Lipids, Fatty Acids, and Trace Elements in the Liver and Muscle of Wild Saithe Pollachius virens

Saithe Pollachius virens are attracted to uneaten salmon feed underneath cages at open-cage salmon farms in Norway. The aggregated Saithe have modified their feeding habits as they have switched from wild prey to uneaten food pellets, which could lead to physiological and biochemical changes in the Saithe. Variations in profiles of total lipids, fatty acids, and trace elements in Saithe liver and muscle were measured to evaluate the influence of fish feed from salmon farms on wild Saithe populations. Farm-aggregated Saithe had higher fat content in liver tissues than did individuals captured more than 25 km away from farms, but no clear differences were found in muscle tissues. High proportions of fatty acids of terrestrial origin, such as oleic, linoleic, and linolenic acids, in liver and muscle tissues of farm-aggregated Saithe reflected the presence of wild Saithe at farms. Accordingly, low proportions of arachidonic, eicosapentaenoic, and docosahexaenoic acids in Saithe tissues mirrored the feeding activity at farms. Variations in specific trace element signatures among fish groups also revealed the farming influence on wild Saithe. High levels of Fe, As, Se, Zn, and B in liver, but also As, B, Li, Hg, and Sr in muscle of Saithe captured away from farms indicated the absence of feeding at farms.This study was part of the project “Evaluation of actions to promote sustainable coexistence between salmon culture and coastal fisheries – ProCoEx” funded by The Norwegian Seafood Research Fund (FHF). The study was also supported by the Norwegian Research Council through the EcoCoast project

The role of the land-surface model for climate change projections over the Iberian Peninsula

The importance of land-surface processes within Regional Climate Models for accurately reproducing the present-day climate is well known. However, their role when projecting future climate is still poorly reported. Hence, this work assesses the influence of the land-surface processes, particularly the contribution of soil moisture, when projecting future changes for temperature, precipitation and wind over a complex area as the Iberian Peninsula, which, in addition, shows great sensitivity to climate change. The main signals are found for the summer season, when the results indicate a strengthening in the increases projected for both mean temperature and temperature variability as a consequence of the future intensification of the positive soil moisture-temperature feedback. The more severe warming over the inner dry Iberian Peninsula further implies an intensification of the Iberian thermal low and, thus, of the cyclonic circulation. Furthermore, the land-atmosphere coupling leads to the projection of a wider future daily temperature range, since maximum temperatures are more affected than minima, a feature absent in non-coupled simulations. Regarding variability, the areas where the land-atmosphere coupling introduces larger changes are those where the reduction in the soil moisture content is more dramatic in future simulations, i.e., the so-called transitional zones. As regards precipitation, weaker positive signals for convective precipitation and more intense negative signals for non-convective precipitation are obtained as a result of the soil moisture-atmosphere interactions. These results highlight the crucial contribution of soil moisture to climate change projections and suggest its plausible key role for future projections of extreme events

Overview of recent TJ-II stellarator results

The main results obtained in the TJ-II stellarator in the last two years are reported. The most important topics investigated have been modelling and validation of impurity transport, validation of gyrokinetic simulations, turbulence characterisation, effect of magnetic configuration on transport, fuelling with pellet injection, fast particles and liquid metal plasma facing components. As regards impurity transport research, a number of working lines exploring several recently discovered effects have been developed: the effect of tangential drifts on stellarator neoclassical transport, the impurity flux driven by electric fields tangent to magnetic surfaces and attempts of experimental validation with Doppler reflectometry of the variation of the radial electric field on the flux surface. Concerning gyrokinetic simulations, two validation activities have been performed, the comparison with measurements of zonal flow relaxation in pellet-induced fast transients and the comparison with experimental poloidal variation of fluctuations amplitude. The impact of radial electric fields on turbulence spreading in the edge and scrape-off layer has been also experimentally characterized using a 2D Langmuir probe array. Another remarkable piece of work has been the investigation of the radial propagation of small temperature perturbations using transfer entropy. Research on the physics and modelling of plasma core fuelling with pellet and tracer-encapsulated solid-pellet injection has produced also relevant results. Neutral beam injection driven Alfvénic activity and its possible control by electron cyclotron current drive has been examined as well in TJ-II. Finally, recent results on alternative plasma facing components based on liquid metals are also presentedThis work has been carried out within the framework of the EUROfusion Consortium and has received funding from the Euratom research and training programme 2014–2018 under Grant Agreement No. 633053. It has been partially funded by the Ministerio de Ciencia, Inovación y Universidades of Spain under projects ENE2013-48109-P, ENE2015-70142-P and FIS2017-88892-P. It has also received funds from the Spanish Government via mobility grant PRX17/00425. The authors thankfully acknowledge the computer resources at MareNostrum and the technical support provided by the Barcelona S.C. It has been supported as well by The Science and Technology Center in Ukraine (STCU), Project P-507F

Mutation Screening Associated with Chromosome 7 Abnormalities Using Next Generation Whole Exome Sequencing

Abstract Abstract 173 One of the most common karyotypic abnormalities identified in myelodysplastic syndromes (MDS) is monosomy 7 (del7) or deletion of the long arm of chromosome 7 (del7q). The presence of del7/del7q carries a poor prognosis in MDS, MDS/myeloproliferative neoplasms (MPN) and acute myeloid leukemia (AML); the impact of these defects appears similar. Recently, a copy-neutral type of loss of heterozygozity (LOH also referred to as a somatic UPD) has been identified on 7q. Microdeletion on 7q corresponding to the EZH2 locus led to identification of inactivating mutations in this gene, though hemizygous EZH2 mutations are only rarely found and do not fully explain del7/7q pathogenesis. We performed a comprehensive analysis of myeloid neoplasms (N=189), using next generation whole exome sequencing technology, including MDS (N=34), MDS/MPN (N=26) or MPN (N=4) and 124 with AML (both primary and secondary). Among them, LOH7, involving del7/del7q were observed in 17% of cases (N=33). To minimize false positives and focus on the most prevalent/relevant somatic events, we implemented a rational bioanalyitic filtering approach, whereby paired DNA (tumor/CD3 lymphocyte) were sequenced and results aligned using Burrows-Wheeler Aligner and variants detected using GATK pipeline (Best Practice Variant Detection from Broad Institute). We focused on searching for del7/7q linked somatic mutational events involved comparisons of mutations in the area of del7q to cases diploid for this locus. We hypothesized that there may be heterozygous mutations of 7q, which could lead to functional haploinsufficiency that is also a result of del7q (haploinsuffcient theory, heterozygous mutations). Conversely, mutations may be either unique to del7q hemizygous inactivation, or shared between 7q diploid and haploid cases. In total, we found alterations in 12 genes located on chromosome 7 (6% of all alterations found). Using filtering strategies we narrowed the focus to “tier 1” mutations to avoid false positives; 11 mutated genes were found in cases with del7/7q and 2 in UPD7q. For example, novel hemzygous (but not heterozygous mutations) of an E3 ubiquitin ligase CUL1 gene were detected only in cases with del7/7q, suggesting that the wild type allele is protective. In cases with diploid 7q, 24 heterozygous alterations were observed (10 genes shared with del7/7q). The previously described EZH2 mutations were seen in heterozygous, homozygous and hemizygous configurations, but were most common in UPD7q (100%), while only 7% of del7/7q cases were positive. Notably, 5/12 mutant genes were located in commonly deleted regions (CDRs) either 7q22, 7q34 or 7q35–36. These CDRs also contain recurrently mutated lesions, including 7q22 (CUX1:n=4; STAG3:n=2), 7q34 (a splicing factor; LUC7L2: n=3) and 7q35–36 (EZH2: n=10). When we investigated the association between haploinsufficiency and heterozygous mutations, among those on del7/7q, cases with wild type forms of corresponding genes showed decreased expression. Similarly, such mutations were occasionally present in diploid configuration; here again the wild type cases showed a decreased expression. These findings suggest that mutated genes located in CDRs can be pathogenic due to both haploinsufficiency of WT genes and heterozygous mutations. EZH2 is a good example of such a gene. We also searched accessory genetic events observed on other chromosomes along with del7/7q and UPD7. By SNP-A, there were clear differences among 3 LOH7 groups, in which del7 was more associated with accessory chromosomal defects than cases with UPD7q or del7. Similarly, mutational patterns were specific to each LOH cohort. For example, while well known frequently mutated genes, such as U2AF1, TET2 and TP53, were commonly found in all 3 LOH7 groups, some specific genes, including the CSMD family, were uniquely observed in monosomy 7, not in del7q or UPD7. Similarly, LOH7q was associated with somatic mutations in SETBP1 and RUNX1. In conclusion, we detected several candidate genes that could be associated with del7/7q and UPD7. Some mutations were heterozygous in cases with diploid 7q and correlated with CDRs on del7/7q without mutation. Certain mutations are specifically observed with del7, while others are commonly observed in all categories of LOH7, including EZH2. Moreover, some genes outside of the chromosome 7 were coincidently mutated with LOH7. Disclosures: Makishima: Scott Hamilton CARES Initiative: Research Funding. Maciejewski:NIH: Research Funding; Aplastic Anemia&MDS International Foundation: Research Funding

Recapturing fish escapes from coastal farms in the western Mediterranean Sea: Insights for potential contingency plans

Escape incidents of farmed fish involve economic losses to fish farms, interactions with local fisheries and environmental impacts to coastal ecosystems. More attention should clearly be paid to preventive measures. It is also essential to develop and establish contingency plans in case of escapes, to mitigate potentially negative socioeconomic and environmental impacts. Three mark-and-recapture experiments simulating escape incidents of sea bass (N = 1000 ind.), sea bream (N = 1000 ind.) and meagre (N = 1000 ind.) were carried out at three coastal fish farms located along the Mediterranean Coast of Spain. First, targeted experimental fishing trials in collaboration with artisanal netters were attempted at each location as potential fast-response contingency plans for recapturing escapees. Targeted fishing was successful on meagre (N = 38 ind., CPUE: 2.2 ind 10 m−2 h−1) and sea bream (N = 8 ind., CPUE: 1.3 ind 10 m−2 h−1), while no sea bass were recaptured. Secondly, recaptures reported from local fishermen (professional and recreational) during the study period were also considered. Altogether, total recapture rates were similar among the three species (sea bass: 5.4%; sea bream: 7.1%; meagre: 8.7%), although the spatial and temporal observations of recaptures varied among species. Recreational fishermen were the only contributors, recapturing 54 tagged sea bass (angling: 85%; spear-fishing: 15%) in mainly shallow coastal waters and about three km distance from the fish-farm during the weeks after release. A total of 71 escaped sea bream were recaptured by both recreational and artisanal fishermen, contributing similarly (recapture rates: 47.9% and 40.8% of total, respectively). Most sea bream recaptures were during the first nine days after release (86% of total recaptures), mainly near the farm facilities (95%), mainly by experimental and artisanal netting (38 and 47 individuals respectively) again near the facility. In parallel, an underwater visual census was carried out at coastal locations and Natura 2000 sites in each study area, to assess the presence of escapees in marine habitats of special interest. Neither escaped sea bass nor escaped meagre were observed during underwater surveys, and only three tagged sea bream were found together at artificial reefs. Given that recaptures of tagged fish differed among fish species and fishing techniques following simulated escape incidents at W-Mediterranean coastal facilities, diverse potential contingency plans are here discussed.This research was part of ESCAFEP project, cofunded by the European Fisheries Fund (EFF) and the Biodiversity Foundation, in collaboration with the Spanish Ministry of Agriculture, Food and the Environment

Whole Exome Sequencing to Predict Response to Hypomethylating Agents in MDS

Abstract Abstract 1698 Hypomethylating agents decitabine and azacitidine are standard treatments for myelodysplastic syndromes (MDS). In their use, one hopes to rectify cytopenias and prolong survival by retarding further disease progression. However, individual treatment responses vary from complete remission (CR) to complete refractoriness. In general, at least 4 cycles of therapy are administered prior to assessing response. Thus, patients may have prolonged exposure to ineffective therapy, suffering toxicities without clinical benefit, while alternative and potentially more effective treatments are delayed. Currently, there are no reliable phenotypic or mutational markers for predicting response to hypomethylating agents. Once whole exome sequencing (WES) became available for more routine analysis, we theorized that somatic mutational patterns may help identify patients who would most benefit from these drugs, thereby maximizing response rate by rational patient selection. To pursue this hypothesis, we screened a cohort of 168 patients with MDS who received either azacitidine or decitabine for the presence of somatic mutations. Only those who received sufficient therapy, i.e., completed at least 4 cycles, were selected for outcome analysis. Targeted Sanger sequencing, including a panel of up to 19 genes frequently affected by somatic mutations was performed. For a representative subset of 26 patients (this subset is expanding) of whom there were 15 responders and 11 non-responders, mutational analysis was performed by WES to select target genes for further analysis. WES utilizes paired DNA (tumor vs. CD3+ lymphocytes) to produce raw sequence reads aligned using Burrows-Wheeler Aligner (BWA). Variants are detected using the Broad Institute's Best Practice Variant Detection GATK toolkit. Median age was 68 years (range, 55–85), 50% were female, and MDS subtypes were as follows: RA/RCUD/RARS 13%, RCMD 16%, RAEB-1/2 20%, MDS/MPN & CMML-1/2 31%, and sAML 20%. Response was assessed using IWG 2006 criteria at 4 and 7 months after therapy initiation. Overall response was 48%; rate of CR (including marrow/cytogenetic CR) was 28%, any HI 20%, SD 22%, and no response 29%. The cohort was then dichotomized into “responders” and “non-responders,” with responders classified as those achieving CR or any HI. Baseline patient characteristics were similar between both groups, including average age at treatment initiation, disease subtypes, proportion of abnormal/complex karyotypes, and presence of common cytogenetic aberrations. Overall, the most frequently mutated genes include TET2/IDH1/IDH2, SRSF2, ASXL1, SF3B1, RUNX1, EZH2/EED/SUZ12, SETBP1, CBL, and PPIAF2. The highest rate of refractoriness was noted in mutants of TET2/IDH1/IDH2 (67%), SF3B1 (67%), U2AF1/2 (67%). We also identified several genes whose mutants were few but associated exclusively with refractory disease (100%), including KIT, ZRSR2, PRPF8, LUC7L2. We next applied a recursive partitioning algorithm to construct a decision tree for identifying the most pivotal mutations associated with response: we found mutant CBL and PPFIA2 to be strongly associated with response, whereas mutant U2AF1/2, SF3B1 and PRPF8 were strongly associated with refractoriness. Our final approach was to dichotomize the cohort by the presence/absence of each mutation/group of mutations, and response within mutant vs. wild type cases was compared. Among refractory cases, TET2/IDH1/IDH2 (26%) and SF3B1 (17%) were most frequently mutated; among responders, mutations in RUNX1 (19% vs. 4%]), CBL (14% vs. 0%), SRSF2 (23% vs. 9%), and SETBP1 (18% vs. 4%) were most frequent. When multiple genes were combined in “either-or” fashion, mutation in TET2, SF3B1, PRPF8, or LUCL71 was significantly associated with refractoriness (52%, p=.0287), whereas mutations of RUNX1, CBL, SRSF2, SETBP1, or PPFIA2 mutation was significantly associated with response (86%, p=.0001). Mutational patterns appear to predict response to standard hypomethylating agents. Identification of the most predictive genes could guide development of molecular maker-based selection of patients for hypomethylating agent therapy, but will require ongoing analysis and additional prospective testing for validation. Disclosures: Advani: Genzyme: Honoraria, Research Funding; Immunomedics: Research Funding. Maciejewski:NIH: Research Funding; Aplastic Anemia&MDS International Foundation: Research Funding