Conventional morphology-based identification is commonly used for routine assessment of freshwater ecosystems. However, cost and time efficient techniques such as high-throughput sequencing (HTS) based approaches may resolve the constraints encountered in conducting morphology-based surveys. Here, we characterized stream macroinvertebrate species diversity and community composition via metabarcoding and morphological analysis from environmental samples collected from the Shigenobu River Basin in Ehime Prefecture, Japan. We compared diversity metrics and assessed both approaches' ability to evaluate the relationship between macroinvertebrate community and environmental variables. In total, we morphologically identified 45 taxa (3 families, six subfamilies, 31 genera, and five species) from 8,276 collected individuals from ten study sites. We detected 44 species by metabarcoding, with 35 species collapsed into 11 groups matching the morphologically identified taxa. A significant positive correlation between logged depth (number of HTS reads) and abundance of morphological taxa was observed, which implied that quantitative data can be used for subsequent analyses. Relatively higher estimates of alpha diversity were calculated from the metabarcoding data in comparison to morphology-based data. However, beta diversity estimates between metabarcoding and morphology data based on both incidence and abundance-based matrices were correlated proving that community differences between sampling sites were preserved in the molecular data.Also, both models were significant, but metabarcoding data (93%) explained a relatively higher percentage of variation in the relationship between community composition and the environmental variables than morphological data (91%). Overall, we present both the feasibility and limitations of HTS-driven estimations of taxonomic richness, community composition, and diversity metrics, and that metabarcoding was proven comparable and more sensitive against morphology-based analysis for stream macroinvertebrate biodiversity assessment and environmental monitoring.

Zwicknia Murányi, gen. n. is erected for the Capnia bifrons species group sensu Zhiltzova, 2001 with the description of three new species based on morphology, mating call, and the mitochondrial DNA marker cytochrome c oxidase I: Z. acuta Murányi & Orci, sp. n., Z. kovacsi Murányi & Gamboa, sp. n. and Z. rupprechti Murányi, Orci & Gamboa, sp. n.. Zwicknia bifrons (Newman, 1838) comb. n. is selected as the type species and redescribed. The other three species placed into Zwicknia, gen. n., Z. sevanica (Zhiltzova, 1964) comb. n., Z. tuberculata (Zhiltzova, 1964) comb. n., and Z. turkestanica (Kimmins, 1950) comb. n. are redescribed based only on morphological characters. Comparative morphological studies and newly discovered characters of the genitalia has allowed for the first time a synopsis of the adults of the West Palaearctic and Nearctic genera of Capniidae. Arsapnia Banks, 1897 (type species: A. decepta Banks, 1897 comb. rev.) is removed from synonymy with Capnia Pictet, 1841 with new combinations, Arsapnia arapahoe (Nelson & Kondratieff, 1988) comb. n., A. coyote (Nelson & Baumann, 1987) comb. n., A. pileata (Jewett, 1966) comb. n., A. sequoia (Nelson & Baumann, 1987) comb. n., A. teresa (Claassen, 1924) comb. n., A. tumida (Claassen, 1924) comb. n., and A. utahensis (Gaufin & Jewett, 1962) comb. n. A new sensu stricto diagnosis of Capnia is proposed with comments on the taxa retained in Capnia sensu lato.

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10Conventional morphology-based identification is commonly used for routine assessment of freshwater 11 ecosystems. However, cost and time efficient techniques such as high-throughput sequencing (HTS) based 12 approaches may resolve the constraints encountered in conducting morphology-based surveys. Here, we 13 characterized stream macroinvertebrate species diversity and community composition via metabarcoding and 14 morphological analysis from environmental samples collected from the Shigenobu River Basin in Ehime 15 Prefecture, Japan. We compared diversity metrics and assessed both approaches' ability to evaluate the 16 relationship between macroinvertebrate community and environmental variables. In total, we morphologically 17 identified 45 taxa (3 families, six subfamilies, 31 genera, and five species) from 8,276 collected individuals 18 from ten study sites. We detected 44 species by metabarcoding, with 35 species collapsed into 11 groups 19 matching the morphologically identified taxa. A significant positive correlation between logged depth (number 20 of HTS reads) and abundance of morphological taxa was observed, which implied that quantitative data can 21 be used for subsequent analyses. Relatively higher estimates of alpha diversity were calculated from the 22 metabarcoding data in comparison to morphology-based data. However, beta diversity estimates between 23 metabarcoding and morphology data based on both incidence and abundance-based matrices were 24 correlated proving that community differences between sampling sites were preserved in the molecular data. 25 Also, both models were significant, but metabarcoding data (93%) explained a relatively higher percentage of 26 variation in the relationship between community composition and the environmental variables than 27 morphological data (91%). Overall, we present both the feasibility and limitations of HTS-driven estimations 28 of taxonomic richness, community composition, and diversity metrics, and that metabarcoding was proven 29 comparable and more sensitive against morphology-based analysis for stream macroinvertebrate biodiversity 30 assessment and environmental monitoring. 31 32 community composition 33 42 al., 2016). 43 Conventional morphological analysis is most commonly used in routine monitoring programs evaluating 44 environmental quality changes. However, this is not only time consuming but has serious issues with accuracy, 45 and consistency in the level of taxonomic identification that highly depends on taxonomic expertise (Hajibabaei 46 et al., 2011). Specifically, small organisms such as the larval stages of stream macroinvertebrates frequently 47 used for river biomonitoring are often difficult or impossible to identify at finer taxonomic resolution (e.g., 48 species level) (Sweeny et al., 2011). A promising alternative approach is DNA metabarcoding -a combination 49 of amplicon-based high-throughput sequencing (HTS) analysis and DNA taxonomy (Hebert et al., 2003). High-50 throughput amplicon sequencing can process large number ...

Non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) are emerging worldwide epidemics, projected to become the leading cause of liver transplants. The strongest genetic risk factor for NAFLD/NASH susceptibility and progression is a single-nucleotide polymorphism (SNP) in the patatin-like phospholipase domain-containing 3 gene (PNPLA3), rs738409, encoding the missense mutation I148M. This aminoacidic substitution interferes with the normal remodeling of lipid droplets in hepatocytes. It is also thought to play a key role in promoting liver fibrosis by inhibiting the release of retinol from hepatic stellate cells. Reducing PNPLA3 levels in individuals homozygous for 148M may be an effective treatment for the entire spectrum of NAFLD, based on gene dosage analysis in the human population, as well as the protective effect of another naturally occurring SNP (rs2294918) in PNPLA3 which, when co-inherited, reduces PNPLA3 mRNA levels to 50% and counteracts disease risk. By screening a clinical compound library targeting specific signaling pathways active in primary human hepatocytes, we identified momelotinib, a drug evaluated in clinical trials to treat myelofibrosis, as a potent down-regulator of PNPLA3 expression, across all genotypes. We found that momelotinib treatment yielded >80% reduction in PNPLA3 mRNA in human primary hepatocytes and stellate cells, as well as in vivo via acute and chronic treatment of WT mice. Using a human multilineage 3D spheroid model of NASH homozygous for the PNPLA3 mutant protein, we additionally show that it decreases PNPLA3 mRNA as well as intracellular lipid content. Furthermore, we show that the effects on PNPLA3 coincide with changes in chromatin accessibility within regulatory regions of the PNPLA3 locus, consistent with inhibition occurring at the level of transcription. In addition to its primary reported targets, the JAK kinases, momelotinib inhibits several non-JAK kinases, including ACVR1. Using a combination of targeted siRNA knockdowns and signaling pathway perturbations, we show that momelotinib reduces the expression of the PNPLA3 gene largely through the inhibition of BMP signaling rather than the JAK/STAT pathway. Overall, our work identified momelotinib as a potential NASH therapeutic and uncovered previously unrecognized connections between signaling pathways and PNPLA3. These pathways may be exploited by drug modalities to “tune down” the level of gene expression, and therefore offer a potential therapeutic benefit to a high at-risk subset of NAFLD/NASH patients.