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.

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.

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.

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.

Polymorphism, gigantism, and cannibalism, one stylonychid ciliate (Ciliophora, Hypotricha) to rule them all.

The morphology, ontogenesis, and molecular phylogeny of the polymorphic and cannibalistic giant forming Tetmemena polymorpha n. sp., found in a brackish water sample in South Korea, were investigated. The present species has long been misidentified as "Oxytricha bifaria." The new investigation shows that the species produces three morphologically different morphs. The small morph is bacterivorous and characterized by its small body size and slim body and it is found only in the stationary and decline phases of the culture. The large morph has a wide body, larger oral apparatus, and feeds on small eukaryotes such as yeast cells and small ciliates. It divides very quickly and produces the other two morphs and found in the exponential phase of the cultures. The giant morph is characterized by its huge body and oral apparatus. It feeds on the small morph cells of the same species and other ciliates, and occurs together with the small morph. Phylogenetic analyses based on the 18S rRNA gene sequences show that the new species is placed in a sister subclade to that containing other Tetmemena sequences. Moreover, Tetmemena indica Bharti et al., 2019 nov. stat. is raised to species level based on the differences in the cyst morphology and the dorsal ciliature to the authoritative Tetmemena pustulata population.

CLDN18.2 and 4-1BB bispecific antibody givastomig exerts antitumor activity through CLDN18.2-expressing tumor-directed T-cell activation.

BACKGROUND: Claudin18.2 (CLDN18.2) is a tight junction protein that has been identified as a clinically proven target in gastric cancer. Stimulation of 4-1BB with agonistic antibodies is also a promising strategy for immunotherapy and 4-1BB+ T cells were reported to be present within the tumor microenvironment of patients with gastric cancer. However, hepatotoxicity-mediated by 4-1BB activation was observed in clinical trials of agonistic anti-4-1BB monoclonal antibodies. METHODS: To specifically activate the 4-1BB+ T cells in tumor and avoid the on-target liver toxicity, we developed a novel CLDN18.2×4-1BB bispecific antibody (termed 'givastomig' or 'ABL111'; also known as TJ-CD4B or TJ033721) that was designed to activate 4-1BB signaling in a CLDN18.2 engagement-dependent manner. RESULTS: 4-1BB+ T cells were observed to be coexisted with CLDN18.2+ tumor cells in proximity by multiplex immunohistochemical staining of tumor tissues from patients with gastric cancer (n=60). Givastomig/ABL111 could bind to cell lines expressing various levels of CLDN18.2 with a high affinity and induce 4-1BB activation in vitro only in the context of CLDN18.2 binding. The magnitude of T-cell activation by givastomig/ABL111 treatment was closely correlated with the CLDN18.2 expression level of tumor cells from gastric cancer patient-derived xenograft model. Mechanistically, givastomig/ABL111 treatment could upregulate the expression of a panel of pro-inflammatory and interferon-γ-responsive genes in human peripheral blood mononuclear cells when co-cultured with CLDN18.2+ tumor cells. Furthermore, in humanized 4-1BB transgenic mice inoculated with human CLDN18.2-expressing tumor cells, givastomig/ABL111 induced a localized immune activation in tumor as evident by the increased ratio of CD8+/regulatory T cell, leading to the superior antitumor activity and long-lasting memory response against tumor rechallenge. Givastomig/ABL111 was well tolerated, with no systemic immune response and hepatotoxicity in monkeys. CONCLUSIONS: Givastomig/ABL111 is a novel CLDN18.2×4-1BB bispecific antibody which has the potential to treat patients with gastric cancer with a wide range of CLDN18.2 expression level through the restricted activation of 4-1BB+ T cells in tumor microenvironment to avoid the risk of liver toxicity and systemic immune response.

Diophrys pauciciliata n. sp. (Ciliophora, Euplotida), a new marine spirotrich ciliate with highly static morphometrics.

A new marine ciliate was discovered from intertidal mudflat from South Korea. Based on the observation of living cells, protargol-impregnated specimens and SEM, we describe it as new species belonging to the genus Diophrys Dujardin, 1841, namely Diophrys pauciciliata n. sp. Its nuclear small subunit ribosomal RNA gene also supports the taxonomic assignment. Diophrys pauciciliata n. sp. is one of the smallest species in the genus. Besides, the bipartite adoral zone and oligomerized dorsal bristles easily discriminate the new species from other congeners. Interestingly, D. pauciciliata n. sp. showed highly static states in all morphometrics (<8.5 % of coefficient of variation) and in particular, the number of frontal adoral membranelles and the counts of dorsal ciliature (e.g., number of dorsal kineties and numbers of dikinetids within each kinety) are invariable. Considering morphometric data of previous studies in the genus Diophrys, the narrow range of character states is unexpected because cell length is usually positively correlated with the numbers of adoral membranelles and dorsal dikinetids.

Morphology and molecular phylogeny of Keronopsis koreana n. sp. (Ciliophora, Hypotricha), a new soil species from South Korea.

The morphology and molecular phylogeny of a new soil ciliate, Keronopsis koreana n. sp., which was discovered in soil from Jindo-gun, South Korea, were investigated. The new species is characterized by the following morphological features: cell more or less ellipsoid or ovoid, both ends broadly rounded; cortical granules irregularly distributed, colorless, 1 µm across; 35-54 adoral membranelles; 12-23 cirri in frontal corona; two frontal and two frontoventral cirral rows; two or three buccal cirri; 23-38 left and 22-37 right marginal cirri; three bipolar dorsal kineties; and two macronuclear nodules and three to five micronuclei. Phylogenetic analyses show that Keronopsis koreana n. sp. is placed within a clade containing K. helluo (type species of Keronopsis) and species belonging to Paraholosticha, Uroleptoides, Parabistichella, Orthoamphisiella, Lamtostyla, and Bistichella in non-dorsomarginalian hypotrichs. Also, the sequences belonging to the Keronopsidae did not cluster together. In addition, a new combination is provided: Parabistichella tasmaniensis (Blatterer and Foissner, 1988) n. comb. (original combination: Keronopsis tasmaniensis).

Morphology and Molecular Phylogeny of Two New Trachelophyllid Ciliates, Monolamellophrya terricola gen. nov., sp. nov. and Trachelophyllum parapiculatum sp. nov. (Litostomatea, Haptoria), From South Korea.

The morphology and molecular phylogeny of two new ciliates, Monolamellophrya terricola gen. nov., sp. nov. and Trachelophyllum parapiculatum sp. nov., discovered in South Korea, were investigated. The two species belong to the suborder Trachelophyllina, which is characterized by the presence of a mucilaginous layer containing lepidosomes covering the cortex. Monolamellophrya terricola gen. nov., sp. nov. is characterized by the presence of a single layer of type II lepidosomes, representing a new genus. Trachelophyllum parapiculatum sp. nov. has only type I lepidosomes covering the cortex, a generic character of the genus Trachelophyllum, and is distinguished from other congeners by a combination of morphological features, including the 15-24 µm long rod-shaped extrusomes, the 9-13 ciliary rows, the 7-11 and 17-25 dikinetids in brush rows 1 and 2, respectively, and the bipolar brush row 3. Furthermore, the 18S rRNA gene sequences of the two new species were provided. The phylogenetic analyses show that the sequence of M. terricola gen. nov., sp. nov. clusters with two other trachelophyllid sequences, and the sequence of T. parapiculatum sp. nov. is placed at the base of these three sequences with full support. Furthermore, the four trachelophyllid sequences that are available so far form a monophyletic clade.

A new "flagship" ciliate, Pseudostylonychia obliquocaudata n. gen., n. sp. (Ciliophora, Hypotricha), from South Korea.

The morphology, ontogenesis, and molecular phylogeny of a new ciliate, Pseudostylonychia obliquocaudata n. gen., n. sp., discovered in a semiterrestrial habitat in South Korea, were investigated. The new species is characterized by an obovate or triangular body with pointed posterior end. The caudal area is distinctly obliquely truncated rightwards. Pseudostylonychia n. gen. is established and characterized by the obovate or triangular body, Stylonychia-like infraciliature and macronuclear pattern, and undulating membranes in Laurentiella pattern. It is also characterized by the involvement of cirri III/2 and IV/3 in primordia formation. SEM investigations show that the resting cyst has high, spirally arranged ridges. Phylogenetic analyses based on SSU rRNA gene sequences show that the new species is placed as a sister to the clade containing sequences of Stylonychia mytilus complex, Stylonychia (Metastylonychia), and Coniculostomum. Furthermore, Stylonychia koreana Kumar et al., 2016 nov. stat. is raised to species level based on the differences in the cyst morphology between the two subspecies of S. ammermanni. Also, Metastylonychia Kumar and Foissner, 2017 nov. stat. is raised to genus rank based on the new molecular data and the morphological and ontogenetic differences to Coniculostomum and Stylonychia.

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).

Factors associated with the effectiveness of part-time patching for intermittent exotropia in children.

PURPOSE: To evaluate the factors associated with the efficacy of low-dose part-time patching in children with intermittent exotropia (IXT). METHODS: In this prospective observational study, we enrolled 186 patients diagnosed with IXT. Outcome measures included office based control scales, magnitude of exo-deviation, and stereoacuity at near and distance after daily patching for 2 h. We analyzed the clinical data and demographic factors association with improvement of IXT. RESULTS: The study was completed by 152 subjects of total enrolled patients on a consecutive basis followed up for 1 year. Decrease in the magnitude of exo-deviation, improvement of control, and or gain of stereoacuity were observed in 31.6% patients of the recruited subjects after part-time patching. Multivariate analyses showed that prognostic factors determining improvement to part-time patching included convergence insufficiency (CI) type IXT (p = 0.016), poor distance stereopsis (p = 0.044), and large exotropic deviation at distance (p = 0.025). CONCLUSIONS: CI-type exotropia, large distance magnitude of exo-deviation, or poor distance stereopsis appear to be associated with a better response to part-time patching. Therefore low dose part-time patching may be a useful non-surgical treatment alternative to delay surgery in these cases.

Trophic Enrichment Factors of Carbon and Nitrogen Isotopic Ratios (Δ13C and Δ15N) in Four Marine Ciliates.

Understanding the magnitude and causes of isotopic fractionation between organisms and their dietary resources is crucial for gaining knowledge on stable isotope ecology. However, little is known regarding the diet-tissue fractionation values of marine ciliates, which play a critical role in the reconstruction of microbial food webs. In the present study, we conducted experiments on two benthic (Pseudokeronopsis pararubra and Protocruzia labiata) and two pelagic (Strombidium sulcatum and Uronemella filificum) marine ciliates, where they were fed with isotopically constant foods (Chaetoceros calcitrans and Isochrysis galbana) under laboratory culture conditions to determine their carbon and nitrogen isotopic fractionation values (Δ13C and Δ15N). The stable isotope values (δ13C and δ15N) of ciliates for all experiments rapidly increased after the initial feeding, with half-lives ranging from 6.1 to 23.0h for δ13C and from 3.1 to 24.9h for δ15N. The Δ13C and Δ15N for all ciliates represented significantly positive enrichments, with overall mean fractionations of 0.6±0.2 and 1.2±0.4, respectively. Irrespective of the dietary type, both Δ13C and Δ15N were very similar for the same ciliate species. These results suggest that Δ13C and Δ15N for marine ciliates are similar to those found in common marine organisms with very little food-dependent variation. Overall, quantifying the specific isotopic fractionation of marine ciliates is expected to provide fundamental information on the trophic transfer of carbon, nitrogen, and energy flow through the microbial pathway in marine ecosystems.

Tunicothrix halophila n. sp., a Secondarily Oligomerized Parabirojimid Hypotrich (Ciliophora, Spirotrichea) From Hypersaline Costal Water in Korea.

Tunicothrix halophila n. sp. was discovered in a hypersaline marine sample from Jeju Island, Korea. It is characterized by the highly reduced number of dorsal bristles. In addition, the main character of the genus Tunicothrix (e.g., alveolar layer) is absent/indistinct. To figure out its identity and phylogenetic relationship, we examined the new species based on modern morphological methods and molecular phylogenetic analyses. Since the parabirojimids are of basal position to core hypotrichs and a smaller data set could show incorrect phylogenetic relationships among the hypotrichs, we used a huge data set composed of 1,460 DNA sequences to infer the phylogenetic tree. The reduction of dorsal bristles is very likely a secondarily evolved character in hypotrichs, resulting in the independent phenotypic adaptation in the hypersaline ecosystems as shown in other hypersaline hypotrichs. Furthermore, the so-called right marginal row 1 in other congeners is found to produce a pretransverse and transverse cirrus and thus we recommend using the term frontoventral row. Based on our data, we can justify Tunicothrix halophila n. sp. as a new species; however, despite the phenotypic distinctiveness, we refrain to establish a new genus because of the missing data and the non-monophyly of Tunicothrix.

Novel anti-4-1BB×PD-L1 bispecific antibody augments anti-tumor immunity through tumor-directed T-cell activation and checkpoint blockade.

BACKGROUND: Stimulation of 4-1BB with agonistic antibodies is a promising strategy for improving the therapeutic efficacy of immune checkpoint inhibitors (ICIs) or for overcoming resistance to ICIs. However, dose-dependent hepatotoxicity was observed in clinical trials with monoclonal anti-4-1BB agonistic antibodies due to the activation of 4-1BB signaling in liver resident Kupffer cells. METHODS: To avoid this on-target liver toxicity, we developed a novel bispecific antibody (4-1BB×PD-L1 bispecific antibody, termed "ABL503") uniquely designed to activate 4-1BB signaling only in the context of PD-L1, while also blocking PD-1/PD-L1 signaling. RESULTS: Functional evaluation using effector cells expressing both 4-1BB and PD-1 revealed superior biological activity of ABL503 compared with the combination of each monoclonal antibody. ABL503 also augmented T-cell activation in in vitro assays and further enhanced the anti-PD-L1-mediated reinvigoration of tumor-infiltrating CD8+ T cells from patients with cancer. Furthermore, in humanized PD-L1/4-1BB transgenic mice challenged with huPD-L1-expressing tumor cells, ABL503 induced superior anti-tumor activity and maintained an anti-tumor response against tumor rechallenge. ABL503 was well tolerated, with normal liver function in monkeys. CONCLUSION: The novel anti-4-1BB×PD-L1 bispecific antibody may exert a strong anti-tumor therapeutic efficacy with a low risk of liver toxicity through the restriction of 4-1BB stimulation in tumors.

Diagnostic yield of targeted next-generation sequencing in infantile nystagmus syndrome.

Background: Infantile nystagmus syndrome (INS) is a genetically heterogeneous disorder. Identifying genetic causes of INS would help clinicians to facilitate clinical diagnosis and provide appropriate treatment. The aim of this study was to determine the diagnostic utility of targeted next-generation sequencing (NGS) for INS.Materials and methods: We recruited 37 patients who were referred to the Neuro-ophthalmology clinics for evaluations of INS. NGS was performed using a targeted panel that included 98 candidate genes associated with INS. We identified pathogenic variants according to guidelines of the American College of Medical Genetics and Genomics. We also calculated the sensitivity and specificity of each clinical sign to assess the diagnostic yield of our gene panel.Results: After variant filtering, annotation, and interpretation, the potential pathogenic variants were detected in 13 of the 37 patients, achieving a molecular diagnostic rate of 35%. The identified genes were PAX6 (n = 4), FRMD7 (n = 4), GPR143 (n = 2), CACNA1F (n = 1), CNGA3 (n = 1) and GUCY2D (n = 1). In approximately 30% (n = 4) of the patients, the initial clinical diagnosis was revised after a molecular diagnosis was performed. The presence of a family history had the highest predictive power for a molecular diagnosis (sensitivity = 61.5%, specificity = 91.7%), and the sensitivity increased when the family history was considered together with one of two clinical signs such as pendular nystagmus waveforms or anterior segment dysgenesis.Conclusions: Our study shows that targeted NGS can be useful to determine a molecular diagnosis for patients with INS. Targeted NGS also helps to confirm a clinical diagnosis in atypical phenotypes or unresolved cases.

Molecular phylogeny of a new gonostomatid ciliate revealing a discrepancy between interphasic and cell divisional patterns (Ciliophora, Hypotricha).

The morphology, ontogenesis, and molecular phylogeny of a new ciliate, Gonostomum koreanum n. sp., discovered in a terrestrial moss sample from South Korea, were investigated. Morphologically, it is characterized by a gonostomatid oral apparatus, two macronuclear nodules, six frontoventral rows, the two rightmost of which (frontoventral rows V and VI) extend posteriorly to near pretransverse and transverse cirri, and three dorsal kineties each with a single caudal cirrus posteriorly. The new species is easily confused with members of the genus Metagonostomum because of the long frontoterminal cirral row but differs mainly in the de novo (vs. intrakinetal) origin of anlage VI, a character found only in Gonostomum and Paragonostomum. To solve the discrepancy between the interphasic and ontogenetic patterns, we additionally performed morphological and multigene analyses on three gonostomatid species, namely Gonostomum koreanum n. sp. and its morphologically (M. gonostomoida) and ontogenetically (G. kuehnelti) most similar species. The multigene analyses show that the new species is closely related to G. kuehnelti and the core gonostomatids consists of five groups based on the origin of the frontoventral rows.