Hierarchical 3D Electrode Design with High Mass Loading Enabling High-Energy-Density Flexible Lithium-Ion Batteries.

Flexible lithium-ion batteries (LIBs) have attracted significant attention owing to their ever-increasing use in flexible and wearable electronic devices. However, the practical application of flexible LIBs in devices has been plagued by the challenge of simultaneously achieving high energy density and high flexibility. Herein, a hierarchical 3D electrode (H3DE) is introduced with high mass loading that can construct highly flexible LIBs with ultrahigh energy density. The H3DE features a bicontinuous structure and the active materials along with conductive agents are uniformly distributed on the 3D framework regardless of the active material type. The bicontinuous electrode/electrolyte integration enables a rapid ion/electron transport, thereby improving the redox kinetics and lowering the internal cell resistance. Moreover, the H3DE exhibits exceptional structural integrity and flexibility during repeated mechanical deformations. Benefiting from the remarkable physicochemical properties, pouch-type flexible LIBs using H3DE demonstrate stable cycling under various bending states, achieving a record-high energy density (438.6 Wh kg-1 and 20.4 mWh cm-2 ), and areal capacity (5.6 mAh cm-2 ), outperforming all previously reported flexible LIBs. This study provides a feasible solution for the preparation of high-energy-density flexible LIBs for various energy storage devices.

LAG-3xPD-L1 bispecific antibody potentiates antitumor responses of T cells through dendritic cell activation.

Several preclinical studies demonstrate that antitumor efficacy of programmed cell death-1 (PD-1)/programmed death-ligand 1 (PD-L1) blockade can be improved by combination with other checkpoint inhibitors. Lymphocyte-activation gene 3 (LAG-3) is an inhibitory checkpoint receptor involved in T cell exhaustion and tumor immune escape. Here, we describe ABL501, a bispecific antibody targeting LAG-3 and PD-L1 in modulating immune cell responses against tumors. ABL501 that efficiently inhibits both LAG-3 and PD-L1 pathways enhances the activation of effector CD4+ and CD8+ T cells with a higher degree than a combination of single anti-LAG-3 and anti-PD-L1. The augmented effector T cell responses by ABL501 resulted in mitigating regulatory-T-cell-mediated immunosuppression. Mechanistically, the simultaneous binding of ABL501 to LAG-3 and PD-L1 promotes dendritic cell (DC) activation and tumor cell conjugation with T cells that subsequently mounts effective CD8+ T cell responses. ABL501 demonstrates its potent in vivo antitumor efficacy in a humanized xenograft model and with knockin mice expressing human orthologs. The immune profiling analysis of peripheral blood reveals an increased abundance of LAG-3hiPD-1hi memory CD4+ T cell subset in relapsed cholangiocarcinoma patients after gemcitabine plus cisplatin therapy, which are more responsive to ABL501. This study supports the clinical evaluation of ABL501 as a novel cancer immunotherapeutic, and a first-in-human trial has started (NCT05101109).

Morphology, Morphogenesis, and Molecular Phylogeny of Neobakuella aenigmatica n. sp. (Ciliophora, Spirotrichea, Bakuellidae).

The morphology and morphogenesis of a new ciliate species, Neobakuella aenigmatica n. sp., which was discovered in an estuary in Korea, were investigated, using live observation, protargol impregnation, and scanning electron microscopy. This new species is characterized by a large (185-300 × 55-105 µm in vivo), elongate-ellipsoidal, flexible but not contractile body. It has ellipsoidal, yellowish cortical granules, 1.3 × 1.0 µm in size. The species has invariably 3 frontal and 2 frontoterminal cirri, about 5-10 buccal and 1-6 parabuccal cirri, 7 midventral rows, and 1 right and 2-4 left marginal rows. The outer left marginal row(s) consists of 1-7 short rows of cirri. The nuclear apparatus comprises 130 macronuclear nodules and 2 spherical micronuclei on average. The dorsal ciliature consists of 3 dorsal kineties. The leftmost left marginal row(s) likely develops from anlagen originating from both the rightmost and leftmost left marginal row(s). The molecular phylogenetic tree based on SSU rDNA suggests the nonmonophyly of the genus Neobakuella.

Phylogeny and Genetic/Morphological Variation of Strombidinopsis minima-like Species (Ciliophora: Choreotrichia).

Six isolates of mineral-enveloped Strombidinopsis minima-like species were collected from the coastal waters across several regions in Korea. Morphological observations and molecular analyses were performed. The ribosomal DNA sequences (including small subunit ribosomal DNA, internal transcriber spacer 1-5.8S ribosomal DNA-internal transcriber spacer 2; and part of large subunit ribosomal DNA) of these six isolates were compared. Their morphological characteristics were also compared with those of S. minima populations reported. The marked genetic differences (with a similarity range of 96.85-98.48%) in SSU rDNA among these S. minima-like entities suggest the existence of multiple species. This finding is also supported by morphological variations detected in this study and reported in the literature (e.g. 15-32 collar membranelles in different populations). In addition, S. minima-like species are clustered with S. batos and S. sinicum, and therefore, our SSU rDNA results support previous results suggesting that the genus Strombidinopsis is not monophyletic in origin. Further collection of morphological and molecular data may facilitate the determination of a new genus carrying mineral-enveloped Strombidinopsis species.

A study on the non-monophyletic genera Australothrix and Holostichides based on multigene and morphological analyses with a reexamination of type materials (Protozoa: Ciliophora).

In a study of ciliate diversity, we collected two new species of the genera Australothrix and Holostichides. Based on nuclear ribosomal genes, our study shows that these two genera are genetically non-monophyletic. To clarify the issues of the non-monophyly, we reexamined type material of H. heterotypicus, H. terrae, and Birojimia soyaensis. Based on multigene and morphometric analyses, Australothrix and Holostichides are clearly non-monophyletic even in mitochondrial CO1 gene trees. The multigene analyses show a clade composed of A. lineae sp. nov., H. terrae, and B. soyaensis, suggesting that a cytopharynx with argyrophilic structures might be their synapomorphy. A list of species with this type of cytopharynx is provided for further studies to either accept or reject this hypothesis. Based on the reexamination, we discriminate the morphologically nearly identical (cryptic/sibling) species H. obliquocirratus sp. nov. from H. heterotypicus and they show significant genetic dissimilarities in the multigene trees. They showed only few morphological (non-quantitative) differences and thus distinguishing them morphologically needs careful investigation.

Morphology and Molecular Phylogeny of Two New Terrestrial Ciliates, Australocirrus rubrus n. sp. and Notohymena gangwonensis n. sp. (Ciliophora: Oxytrichidae), from South Korea.

Two new soil oxytrichids, Australocirrus rubrus n. sp. and Notohymena gangwonensis n. sp., were discovered from South Korea. Morphologically, A. rubrus shares many features with A. australis, and these two species form a single clade in a molecular tree based on nuclear small subunit ribosomal RNA (SSU rRNA) gene sequences. Australocirrus rubrus mainly differs from A. australis in the color (citrine color vs. reddish) and distribution of the cortical granules. Additionally, we confirm that the genus Australocirrus is not a monophyletic group, as A. shii is separated from the clade comprising the other Australocirrus species, being clustered instead with other taxa. Notohymena gangwonensis n. sp. mainly differs from its congeners by the following combination of features: irregularly distributed cortical granules (vs. arranged in groups associated with cirri and dorsal kineties), variable four or five (usually four) transverse cirri (vs. invariable five), and the anteriormost pretransverse cirrus V/2 on 13.2-16.1% of cell length (vs. on or above 18.9% of cell length). Currently, there are no available gene sequences for members of the genus Notohymena, thus we provide SSU rRNA gene sequences from the new species of Notohymena, as well as detailed morphological descriptions of the novel species.

Morphology and Phylogeny of the Soil Ciliate Parabistichella multilineae sp. nov. (Protozoa: Ciliophora: Hypotricha).

The morphology and molecular phylogeny of Parabistichella multilineae sp. nov., which was discovered in soil from Gagye Beach, South Korea, were investigated. The new species is characterized as follows: body size about 200 × 47 µm in vivo, slenderly lanceolate and twisted; two ellipsoidal macronuclei and usually two globular micronuclei; two types of cortical granules; four or five frontoventral rows, of which rows 1 and 2 are conspicuously developed and rows 3 and 4 are fully extended; and caudal cirri are lacking. Phylogenetic analyses based on the SSU rDNA sequences showed that the new species clusters in a soft polytomy with P. variabilis and Uroleptoides magnigranulosus.

Morphology and Molecular Phylogeny of Two New Ciliates, Holostichides heterotypicus n. sp. and Holosticha muuiensis n. sp. (Ciliophora: Urostylida).

Two new urostylid species, Holostichides heterotypicus n. sp. and Holosticha muuiensis n. sp., were discovered in South Korea. Morphological and phylogenetic analyses were carried out to confirm that these species are new to science. Holostichides heterotypicus is mainly characterized by the following combination of features: 110-205 µm long in vivo; 5-10 frontoterminal cirri; 6-8 midventral pairs with 2-3 midventral cirral rows; cortical granules present; four bipolar dorsal kineties; and 6-9 caudal cirri. Ontogenetic features of H. heterotypicus are similar to those of H. typicus. Phylogenetic analyses revealed that H. heterotypicus was distantly separated from bakuellid genera Apobakuella, Bakuella, Metaurostylopsis, and Neobakuella. This result is supported by the following features: transverse cirri (present in the other four bakuellids vs. absent in Holostichides) and caudal cirri (absent in the other four bakuellids vs. present in Holostichides). Holosticha muuiensis n. sp. is mainly distinguished from its congeners by the following combination of features: 100-185 long in vivo; shortened undulating membrane; cortical granules lacking; contractile vacuole absent; 51-66 adoral zone of membranelles; 42-60 macronuclear nodules; and five bipolar dorsal kineties. In the phylogenetic tree, Holosticha muuiensis n. sp. clustered with a Holosticha group (containing Holosticha diademata, Holosticha foissneri, and Holosticha heterofoissneri).

Pseudonotohymena antarctica n. g., n. sp. (Ciliophora, Hypotricha), a New Species from Antarctic Soil.

A new soil ciliate, Pseudonotohymena antarctica n. g., n. sp., from King George Island, Antarctica, is described based on live observation, protargol impregnation, and its 18S rRNA gene. The new genus Pseudonotohymena is morphologically similar to the genus Notohymena Blatterer and Foissner in the following characteristics: 18 fronto-ventral-transverse cirri, a flexible body, undulating membranes, dorsomarginal kineties, and the number of cirri in the marginal rows. However, Pseudonotohymena differs from Notohymena particularly in the dorsal ciliature, that is, in possessing a nonfragmented dorsal kinety (vs. fragmented). In addition, the molecular phylogenetic relationship of the new species differs from that of Notohymena species. On the basis of the morphological features, the genetic data, and morphogenesis, we establish P. antarctica n. g., n. sp. In addition, the cyst morphology of this species is described.

Euduboscquella costata n. sp. (Dinoflagellata, Syndinea), an Intracellular Parasite of the Ciliate Schmidingerella arcuata: Morphology, Molecular Phylogeny, Life Cycle, Prevalence, and Infection Intensity.

The syndinean dinoflagellate Euduboscquella costata n. sp., an intracellular parasite of the tintinnid ciliate Schmidingerella arcuata, was discovered from Korean coastal water in November of 2013. Euduboscquella costata parasitized in about 62% of the host population, with infection intensity (= number of trophonts in a single host cell) ranging from 1 to 8. Based on morphology and nuclear 18S ribosomal RNA gene sequences, the parasite is new to science. Euduboscquella costata n. sp. had an infection cycle typical of the genus, but had morphological and developmental features that distinguished it from congeneric species. These features include: (1) episome of the trophont with 25-40 grooves converging toward the center of the shield; (2) a narrow, funnel-shaped lamina pharyngea extending from the margin of the episomal shield to the nucleus; (3) persistence of grooves during extracellular development (sporogenesis); (4) a single food vacuole during sporogenesis; (5) separation of sporocytes early in sporogenesis, regardless of type of spore formed; and (6) dinospore size (ca. 14 µm in length) and shape (bulbous episome with narrower, tapering hyposome). After sporogenesis, E. costata produced four different types of spore that showed completely identical 18S rRNA gene sequences. The gene sequence was completely identical with a previously reported population, Euduboscquella sp. ex S. arcuata, from Assawoman Bay, USA, indicating that the two populations are likely conspecific. Favella ehrenbergii, a widely recorded tintinnid known to host Euduboscquella spp., co-occurred with S. arcuata, but was not infected by E. costata in field samples or during short-term, cross-infection experiments.

The effects of vascular endothelial growth factor (VEGF) on human orbital preadipocyte.

PURPOSE: To investigate the presence of the Vascular Endothelial Growth Factor (VEGF) and its receptor (VEGFR) in human orbital preadipocytes, and to evaluate the effect of VEGF on human orbital preadipocyte differentiation and adipogenesis in vitro. RESULTS: Four isoforms of VEGF (VEGF121, 155, 189, and 206), VEGFR-1, VEGF-2, and neuropilin-1 were expressed in human orbital preadipocytes. Treatment with 100 ng/ml VEGF induced higher expressions of C/EBPα and LPL than the non-treated control (p = 0.03 and p = 0.01) or treatment with 50ng/ml (p = 0.04 for both). At both concentrations VEGF enhanced the accumulation of intra-cytoplasmic lipid versus the control, and treatment with 100 ng/ml VEGF induced more lipid accumulation than treatment with 50 ng/ml VEGF (p = 0.03). CONCLUSIONS: VEGF and VEGFR were observed in human orbital preadipocytes, and exogenous VEGF enhanced adipogenesis in these cells. These results suggest that VEGF plays a role as an autocrine or paracrine growth factor during human orbital preadipocyte differentiation.

Morphology and Molecular Phylogeny of Pseudocyrtohymena koreana n. g., n. sp. and Antarctic Neokeronopsis asiatica Foissner et al., 2010 (Ciliophora, Sporadotrichida), with a Brief Discussion of the Cyrtohymena Undulating Membranes Pattern.

We discovered a new brackish water oxytrichid Pseudocyrtohymena koreana n. g., n. sp. in South Korea and investigated the new species on the basis of morphology, ontogenesis, and 18S rRNA gene sequences. The new genus has the 18 frontal-ventral-transverse cirri of typical oxytrichids with flexible body, cortical granules, Cyrtohymena undulating membranes (UM), and one left and one right marginal cirral row. Ontogenesis of the new species indicated that dorsal kinety anlage 3 stretches within the parental row without any fragmentations (Urosomoida pattern) and exclusively forms all caudal cirri. The new genus is morphologically similar to Cyrtohymena Foissner, 1989, but has the following distinctive features: (i) caudal cirri absent in dorsal kineties 1 and 2 (vs. present in Cyrtohymena); and (ii) dorsal kinety 3 nonfragmented (vs. fragmented in Cyrtohymena). Further, we collected an additional species Neokeronopsis asiatica Foissner et al. 2010, from King George Island, Antarctica, and the species shares the morphology of UM with Cyrtohymena. Herein, we describe the previously unidentified characteristics of N. asiatica (i.e., cortical granules, body flexibility, contractile vacuole, and 18S rRNA gene sequence). In addition, we obtained two 18S rRNA gene sequences from Cyrtohymena muscorum and Parasterkiella thompsoni to expand samples for phylogenetic analysis. Our 18S rRNA gene tree supports the hypothesis that the Cyrtohymena UM pattern might have evolved several times in hypotrichs (e.g., Neokeronopsidae, Oxytrichinae, and Stylonychinae).

Morphology and Molecular Phylogeny of Two New Brackish Water Ciliates of Bakuella (Ciliophora: Urostylida: Bakuellidae) from South Korea.

Two new species of Bakuella, B. (B.) incheonensis n. sp. and B. (Pseudobakuella) litoralis n. sp., were discovered in brackish water near Aamdo Shore Park, Incheon, South Korea. We conducted a morphological study based on live observations and protargol-impregnated specimens, and a molecular analysis using nuclear SSU rRNA gene sequences. Bakuella (B.) incheonensis is diagnosed by: body size 70-105 × 20-40 µm in vivo, 21-25 adoral membranelles, three or four frontoterminal cirri, midventral complex composed of 7-10 midventral pairs with one or two rows and terminating at about 62% of body length, 20-28 left and 25-32 right marginal cirri, 58-87 macronuclear nodules, and yellowish cortical granules. Bakuella (Pseudobakuella) litoralis is diagnosed by: body size 90-125 × 30-40 µm in vivo, 25-33 adoral membranelles, 3-5 buccal cirri, midventral complex composed 10-15 midventral pairs with one or two rows and terminated at 70% of body length, one or two pretransverse cirri, 3-6 transverse cirri, 26-39 left and 29-47 right marginal cirri, 49-84 macronuclear nodules, and two types of cortical granules. Molecular phylogeny using SSU rRNA gene sequences shows a nonmonophyletic relationship among Bakuella species and emphasizes the need for further morphogenetic studies of this genus and other related hypotrichs.

Revision of Strombidium paracalkinsi (Ciliophora: Oligotrichea: Oligotrichia), with Comparison of Strombidiids Bearing Thigmotactic Membranelles.

This study analyses the morphological characters of Strombidium paracalkinsi, a species bearing thigmotactic membranelles, collected from Jangmok Bay at the southern coast of Korea. The small subunit ribosomal RNA gene of S. paracalkinsi was sequenced for the first time. By a combination of characters, the thigmotactic membranelles of S. paracalkinsi differ from the thigmotactic membranelles of other strombidiids. They are (1) positioned on dorsal side, (2) contiguous with three membranelles of the adoral zone, but separated by nonciliated portions of the polykinetids, and (3) composed of two rows of kinetids. Except for the thigmotactic membranelles, the morphology of S. paracalkinsi conforms to the diagnosis of the genus Strombidium. Our phylogenetic tree confirms the nonmonophyly of the genus Strombidium. The thigmotactic membranelles are a promising feature for a future split of the genus Strombidium, but require further studies, especially on Strombidium calkinsi Fauré-Fremiet 1932.

Morphology and molecular phylogeny of two hypotrichous ciliates (Ciliophora, Spirotrichea) from South Korea, including Hemiurosomoida koreana n. sp.

The living morphology, infraciliature, and molecular phylogeny of a new soil ciliate, Hemiurosomoida koreana n. sp., discovered in a sample collected from a mountain in the northeast of South Korea, were investigated. The new species possesses the characteristics of the genus Hemiurosomoida, i.e., a reduced number of frontal-ventral-transverse cirri, three dorsal kineties of which kineties 1 and 2 each bears a caudal cirrus, and a single dorsomarginal kinety. It is distinguishable from congeners and other similar species by at least one distinct qualitative or quantitative character including the body size, the presence and arrangement of cortical granules, the number of adoral membranelles, marginal cirri, and dorsal dikinetids, or by the arrangement of transverse cirri. Phylogenetic analyses based on 18S rRNA gene sequences also support the assignment of the new species to the non-monophyletic genus Hemiurosomoida. In addition, the living morphology, infraciliature, and the 18S rRNA gene sequence of a Korean population of Nothoholosticha flava were studied.

Morphology, morphogenesis, and molecular phylogeny of Aspidisca koreana n. sp. (Ciliophora, Euplotida) from South Korea.

The morphology, morphogenesis, and molecular phylogeny of a new ciliate, Aspidisca koreana n. sp., discovered in the eastern coast of South Korea, were investigated. The morphological description is based on the observation of living cells, 4'-6-diamidino-2-phenylindole (DAPI) and silver-stained specimens (e.g., protargol, silver nitrate), and scanning electron micrographs. The new species is characterized by having a small body size (17-25 × 15-18 µm in vivo), a distinct peristomial spur on the posterior portion of left margin, seven frontoventral cirri in "polystyla-arrangement", and the arrangement of the anterior portion of adoral zone of membranelles, i.e., anteriormost membranelle is distinctly separated from the other three membranelles. The morphogenesis follows the typical pattern of this genus. Phylogenetic analyses, using the 18S rDNA sequence, also support the establishment of a new species.

Redescription and molecular phylogeny of Trochilia sigmoides Dujardin, 1841 (Ciliophora, Cyrtophoria) collected from South Korea.

During a survey of Korean marine ciliates, Trochilia sigmoides, the type species of the genus Trochilia, was collected and examined using in vivo observation and protargol impregnation. Moreover, scanning electron microscopy and 18S rRNA gene sequencing have been applied for the first time to study this species. Morphologically, T. sigmoides is characterized by the small body size, the oval body outline, and the spiral dorsal ridges. The Korean population of T. sigmoides shows only minute differences to other populations reported in the literature, mainly in body size and the number of dorsal ridges. Phylogenetic analyses based on 18S rRNA gene sequences show that T. sigmoides and T. petrani are placed together with two members of the family Kyaroikeidae, causing the family Dysteriidae to be non-monophyletic. The present new data increase the knowledge about the morphology and phylogeny of the genus Trochilia and would assist in understanding the phylogenetic relationship between the free-living Dysteriidae and the parasitic Kyaroikeidae.

F-RDW: Redirected Walking With Forecasting Future Position.

In order to serve better VR experiences to users, existing predictive methods of Redirected Walking (RDW) exploit future information to reduce the number of reset occurrences. However, such methods often impose a precondition during deployment, either in the virtual environment's layout or the user's walking direction, which constrains its universal applications. To tackle this challenge, we propose a mechanism F-RDW that is twofold: (1) forecasts the future information of a user in the virtual space without any assumptions by using the conventional method, and (2) fuse this information while maneuvering existing RDW methods. The backbone of the first step is an LSTM-based model that ingests the user's spatial and eye-tracking data to predict the user's future position in the virtual space, and the following step feeds those predicted values into existing RDW methods (such as MPCRed, S2C, TAPF, and ARC) while respecting their internal mechanism in applicable ways. The results of our simulation test and user study demonstrate the significance of future information when using RDW in small physical spaces or complex environments. We prove that the proposed mechanism significantly reduces the number of resets and increases the traveled distance between resets, hence augmenting the redirection performance of all RDW methods explored in this work. Our project and dataset are available at https://github.com/YonseiCGnA-VR/F-RDW.

Anti-4-1BB×PD-L1 Bispecific Antibody Reinvigorates Tumor-Specific Exhausted CD8+ T Cells and Enhances the Efficacy of Anti-PD-1 Blockade.

PURPOSE: To overcome the limited efficacy of immune checkpoint blockade, there is a need to find novel cancer immunotherapeutic strategies for the optimal treatment of cancer. The novel anti-4-1BB×PD-L1 bispecific antibody-ABL503 (also known as TJ-L14B)-was designed to simultaneously target PD-L1 and 4-1BB, and demonstrated strong antitumor T-cell responses without considerable toxicity. Here, we investigated how the combination of ABL503 and anti-PD-1 blockade affected the reinvigoration of exhausted tumor-infiltrating CD8+ T cells (CD8+ TILs) and anti-tumor efficacy. EXPERIMENTAL DESIGN: Single cell suspensions of hepatocellular carcinoma and ovarian cancer from treatment-naive patients were used for immunophenotyping of CD8+ TILs and in vitro functional assays. Humanized hPD-1/hPD-L1/h4-1BB triple knock-in mice were used to evaluate the effects of ABL503 and anti-PD-1 blockade in vivo. RESULTS: We observed that ABL503 successfully restored the functions of 4-1BB+ exhausted CD8+ TILs, which were enriched for tumor-specific T cells but unresponsive to anti-PD-1 blockade. Importantly, compared to anti-PD-1 blockade alone, the combination of ABL503 and anti-PD-1 blockade further enhanced the functional restoration of human CD8+ TILs in vitro. Consistently, the combination of ABL503 with anti-PD-1 in vivo significantly alleviated tumor growth, and induced enhanced infiltration and activation of CD8+ TILs. CONCLUSIONS: ABL503-a PD-L1 and 4-1BB dual-targeting bispecific antibody-elicits pronounced additive tumor growth inhibition, with increased infiltration and functionality of exhausted CD8+ T cells, which in turn enhances the anti-cancer effects of anti-PD-1 blockade. These promising findings suggest that ABL503 (TJ-L14B) in combination with PD-1 inhibitors will likely further enhance therapeutic benefit in clinical trials.

EUKARYOME: the rRNA gene reference database for identification of all eukaryotes.

Molecular identification of micro- and macroorganisms based on nuclear markers has revolutionized our understanding of their taxonomy, phylogeny and ecology. Today, research on the diversity of eukaryotes in global ecosystems heavily relies on nuclear ribosomal RNA (rRNA) markers. Here, we present the research community-curated reference database EUKARYOME for nuclear ribosomal 18S rRNA, internal transcribed spacer (ITS) and 28S rRNA markers for all eukaryotes, including metazoans (animals), protists, fungi and plants. It is particularly useful for the identification of arbuscular mycorrhizal fungi as it bridges the four commonly used molecular markers-ITS1, ITS2, 18S V4-V5 and 28S D1-D2 subregions. The key benefits of this database over other annotated reference sequence databases are that it is not restricted to certain taxonomic groups and it includes all rRNA markers. EUKARYOME also offers a number of reference long-read sequences that are derived from (meta)genomic and (meta)barcoding-a unique feature that can be used for taxonomic identification and chimera control of third-generation, long-read, high-throughput sequencing data. Taxonomic assignments of rRNA genes in the database are verified based on phylogenetic approaches. The reference datasets are available in multiple formats from the project homepage, http://www.eukaryome.org.