A New Distinctive Darwin Wasp Represents the First Record of the Ophion minutus Species-group (Hymenoptera: Ichneumonidae: Ophioninae) from Japan and the Far East, with an Analysis of DNA Barcode-based Species Delimitation in Ophion.

A new Darwin wasp species, Ophion kobensis Shimizu sp. nov. (Hymenoptera: Ichneumonidae: Ophioninae), is described using the integrated morphological and molecular species delimitation approaches. Our results indicate that the new species is closely related to European O. ventricosus Gravenhorst, 1829 of the O. minutus species-group but can be distinguished using morphological characters, such as entirely black body colour with some light-yellow marks and not inclined epicnemial carina in lateral view. This record of the new species represents the first record of O. minutus species-group from Japan and the Far East. Phylogenetic analysis indicate that the O. minutus species-group is weakly recovered as monophyletic and sister to Ophion s. str. The analysis also indicated that two clades within the O. minutus species-group (O. minutus and O. ventricosus) diverged significantly. This suggests that the species-group, as well as the two included clades, could potentially be treated as separate species-groups or genera. The present study supports previous integrative taxonomic and phylogenetic studies of Ophion and represents a first fundamental step for studies focused on the challenging Japanese and Far Eastern Ophion.

Mechanisms of preferential bone formation in myeloma bone lesions by proteasome inhibitors.

Proteasome inhibitors (PIs) can preferentially restore bone in bone-defective lesions of patients with multiple myeloma (MM) who respond favorably to these drugs. Most prior in vitro studies on PIs used continuous exposure to low PI concentrations, although pharmacokinetic analysis in patients has shown that serum concentrations of PIs change in a pulsatile manner. In the present study, we explored the effects of pulsatile treatment with PIs on bone metabolism to simulate in vivo PI pharmacokinetics. Pulsatile treatment with bortezomib, carfilzomib, or ixazomib induced MM cell death but only marginally affected the viability of osteoclasts (OCs) with F-actin ring formation. Pulsatile PI treatment suppressed osteoclastogenesis in OC precursors and bone resorption by mature OCs. OCs robustly enhanced osteoblastogenesis in cocultures with OCs and MC3T3-E1 pre-osteoblastic cells, indicating OC-mediated coupling to osteoblastogenesis. Importantly, pulsatile PI treatment did not impair robust OC-mediated osteoblastogenesis. These results suggest that PIs might sufficiently reduce MM cell-derived osteoblastogenesis inhibitors to permit OC-driven bone formation coupling while suppressing OC differentiation and activity in good responders to PIs. OC-mediated coupling to osteoblastogenesis appears to be a predominant mechanism for preferential occurrence of bone regeneration at sites of osteoclastic bone destruction in good responders.

The Xanthine Oxidase Inhibitor Febuxostat Suppresses Adipogenesis and Activates Nrf2.

Xanthine oxidoreductase (XOR) is a rate-limiting enzyme in purine catabolism that acts as a novel regulator of adipogenesis. In pathological states, xanthine oxidoreductase activity increases to produce excess reactive oxygen species (ROS). The nuclear factor erythroid 2-related factor 2 (Nrf2) is a critical inducer of antioxidants, which is bound and repressed by a kelch-like ECH-associated protein 1 (Keap1) in the cytoplasm. The Keap1-Nrf2 axis appears to be a major mechanism for robust inducible antioxidant defenses. Here, we demonstrate that febuxostat, a xanthine oxidase inhibitor, alleviates the increase in adipose tissue mass in obese mouse models with a high-fat diet or ovariectomy. Febuxostat disrupts in vitro adipocytic differentiation in adipogenic media. Adipocytes appeared at day 7 in absence or presence of febuxostat were 160.8 ± 21.2 vs. 52.5 ± 12.7 (p < 0.01) in 3T3−L1 cells, and 126.0 ± 18.7 vs. 55.3 ± 13.4 (p < 0.01) in 10T1/2 cells, respectively. Adipocyte differentiation was further enhanced by the addition of hydrogen peroxide, which was also suppressed by febuxostat. Interestingly, febuxostat, but not allopurinol (another xanthine oxidase inhibitor), rapidly induced the nuclear translocation of Nrf2 and facilitated the degradation of Keap1, similar to the electrophilic Nrf2 activator omaveloxolone. These results suggest that febuxostat alleviates adipogenesis under oxidative conditions, at least in part by suppressing ROS production and Nrf2 activation. Regulation of adipocytic differentiation by febuxostat is expected to inhibit obesity due to menopause or overeating.

Online citizen sciences reveal natural enemies and new occurrence data of Meteorusstellatus Fujie, Shimizu & Maeto, 2021 (Hymenoptera, Braconidae, Euphorinae).

Background: Citizen science is a research approach that involves collaboration between professional scientists and non-professional volunteers. The utilisation of recent online citizen-science platforms (e.g. social networking services) has greatly revolutionised the accessibility of biodiversity data by providing opportunities for connecting professional and citizen scientists worldwide. Meteorusstellatus Fujie, Shimizu & Maeto, 2021 (Hymenoptera, Braconidae, Euphorinae) has been recorded from the Oriental Islands of Japan and known to be a gregarious endoparasitoid of two macro-sized sphingid moths of Macroglossum, Ma.passalus (Drury) and Ma.pyrrhosticta Butler. It constructs characteristic star-shaped communal cocoons, suspended by a long cable. Although M.stellatus has been reported only from the Oriental Islands of Japan, the authors recognise its occurrence and ecological data from Taiwan and the Palaearctic Island of Japan through posts on online citizen-science groups about Taiwanese Insects on Facebook and an article on a Japanese citizen-scientist's website. New information: Through collaboration between professional and citizen scientists via social media (Facebook groups) and websites, the following new biodiversity and ecological data associated with M.stellatus are provided: Meteorusstellatus is recorded for the first time from Taiwan and the Palaearctic Region (Yakushima Is., Japan).Cechetraminor (Butler, 1875), Hippotioncelerio (Linnaeus, 1758) and Macroglossumsitiene (Walker, 1856) (Lepidoptera, Sphingidae) are recorded for the first time as hosts of M.stellatus and two of which (C.minor and H.celerio) represent the first genus-level host records for M.stellatus.Mesochorus sp. (Hymenoptera, Ichneumonidae), indeterminate species of Pteromalidae and Trichogrammatidae (Hymenoptera), are recognised as hyperparasitoid wasps of M.stellatus.Parapolybiavaria (Fabricius, 1787) (Hymenoptera, Vespidae) is reported as a predator of pendulous communal cocoons of M.stellatus. The nature of suspended large-sized communal cocoons of M.stellatus and the importance and limitations of digital occurrence data and online citizen science are briefly discussed.

TGF-ß-activated kinase-1 inhibitor LL-Z1640-2 reduces joint inflammation and bone destruction in mouse models of rheumatoid arthritis by inhibiting NLRP3 inflammasome, TACE, TNF-α and RANKL expression.

OBJECTIVES: Aberrant NLRP3 inflammasome activation has been demonstrated in rheumatoid arthritis (RA), which may contribute to debilitating inflammation and bone destruction. Here, we explored the efficacy of the potent TGF-ß-activated kinase-1 (TAK1) inhibitor LL-Z1640-2 (LLZ) on joint inflammation and bone destruction in collagen-induced arthritis (CIA). METHODS: LL-Z1640-2 was administered every other day in CIA mice. Clinical and histological evaluation was performed. Priming and activation of NLRP3 inflammasome and osteoclastogenic activity were assessed. RESULTS: NLRP3 inflammasome formation was observed in synovial macrophages and osteoclasts (OCs) in CIA mice. TACE and RANKL were also overexpressed in synovial macrophages and fibroblasts, respectively, in the CIA joints. Treatment with LLZ mitigated all the above changes. As a result, LLZ markedly suppressed synovial hypertrophy and pannus formation to alleviate pain and inflammation in CIA mice. LLZ could block the priming and activation of NLRP3 inflammasome in RAW264.7 macrophage cell line, primary bone marrow macrophages and OCs upon treatment with LPS followed by ATP, thereby suppressing their IL-1ß production. LLZ also suppressed LPS-induced production of TACE and TNF-α in bone marrow macrophages and abolished IL-1ß-induced production of MMP-3, IL-6 and RANKL in synovial fibroblasts. In addition, LLZ directly inhibits RANKL-mediated OC formation and activation. CONCLUSION: TAK1 inhibition with LLZ may become a novel treatment strategy to effectively alleviate inflammasome-mediated inflammation and RANKL-induced osteoclastic bone destruction in joints alongside its potent suppression of TNF-α and IL-6 production and proteinase-mediated pathological processes in RA.

Surgical Orthodontic Treatment of Skeletal Mandibular Protrusion with Multiple Impacted Ankylosed Teeth Treated with Alveolar Corticotomy.

AIM: To present a case of skeletal mandibular protrusion with multiple impacted ankylosed teeth, which were treated with conventional orthognathic surgery and alveolar corticotomy, respectively. BACKGROUND: Tooth ankylosis is characterized by the fusion of a root surface with the surrounding alveolar bone. Various treatment modalities for tooth ankylosis have been developed and are used commonly in the clinic. Corticotomy is defined as the application of intentional surgical injury to the cortical bone to mobilize a tooth with the adjacent bone and soft tissues. The corticotomy technique has been improved in recent years to avoid possible risks such as periodontal damage, tooth devitalization, and osseous necrosis due to an inadequate blood supply. CASE DESCRIPTION: A female patient aged 16 years and 3 months was diagnosed with anterior crossbite and the impaction and ankylosis of multiple canines. After the confirmation of ankylosis, alveolar corticotomy was performed on the maxillary left and mandibular right canines. After 6 months of traction (patient age, 19 years and 7 months), both canines had extruded successfully. After the completion of preoperative treatment, bilateral intraoral vertical ramus osteotomy was performed to correct the anterior crossbite involving a skeletal mandibular protrusion. The amounts of mandibular setback on the right and left sides were approximately 7 and 5 mm, respectively. The total treatment period was 55 months. Acceptable occlusion with a balanced profile was maintained over a 5-year retention period, indicating the long-term stability and success of the treatment. CONCLUSION: Our results indicate that alveolar corticotomy should be considered to facilitate the treatment of multiple impacted ankylosed teeth. CLINICAL SIGNIFICANCE: This report proposes an efficacy of alveolar corticotomy for extrusion of impacted ankylosed teeth.

TAK1 is a pivotal therapeutic target for tumor progression and bone destruction in myeloma.

Along with the tumor progression, the bone marrow microenvironment is skewed in multiple myeloma (MM), which underlies the unique pathophysiology of MM and confers aggressiveness and drug resistance in MM cells. TGF-ß-activated kinase-1 (TAK1) mediates a wide range of intracellular signaling pathways. We demonstrate here that TAK1 is constitutively overexpressed and phosphorylated in MM cells, and that TAK1 inhibition suppresses the activation of NF-κB, p38MAPK, ERK and STAT3 to decrease the expression of critical mediators for MM growth and survival, including PIM2, MYC, Mcl-1, IRF4, and Sp1, along with a substantial reduction in the angiogenic factor VEGF in MM cells. Intriguingly, TAK1 phosphorylation was also induced along with upregulation of vascular cell adhesion molecule-1 (VCAM-1) in bone marrow stromal cells (BMSCs) in cocultures with MM cells, which facilitated MM cell-BMSC adhesion while inducing IL-6 production and receptor activator of nuclear factor κ-Β ligand (RANKL) expression by BMSCs. TAK1 inhibition effectively impaired MM cell adhesion to BMSCs to disrupt the support of MM cell growth and survival by BMSCs. Furthermore, TAK1 inhibition suppressed osteoclastogenesis enhanced by RANKL in cocultures of bone marrow cells with MM cells, and restored osteoblastic differentiation suppressed by MM cells or inhibitory factors for osteoblastogenesis overproduced in MM. Finally, treatment with the TAK1 inhibitor LLZ1640-2 markedly suppressed MM tumor growth and prevented bone destruction and loss in mouse MM models. Therefore, TAK1 inhibition may be a promising therapeutic option targeting not only MM cells but also the skewed bone marrow microenvironment in MM.

Integrative taxonomy and analysis of species richness patterns of nocturnal Darwin wasps of the genus Enicospilus Stephens (Hymenoptera, Ichneumonidae, Ophioninae) in Japan.

The predominantly tropical ophionine genus Enicospilus Stephens, 1835 is one of the largest genera of Darwin wasps (Hymenoptera, Ichneumonidae), with more than 700 extant species worldwide that are usually crepuscular or nocturnal and are parasitoids of Lepidoptera larvae. In the present study, the Japanese species of Enicospilus are revised using an integrative approach (combined morphology and DNA barcoding). On the basis of 3,110 specimens, 47 Enicospilus species are recognised in Japan, eight of which are new species (E. acutus Shimizu, sp. nov., E. kunigamiensis Shimizu, sp. nov., E. limnophilus Shimizu, sp. nov., E. matsumurai Shimizu, sp. nov., E. pseudopuncticulatus Shimizu, sp. nov., E. sharkeyi Shimizu, sp. nov., E. takakuwai Shimizu, sp. nov., and E. unctus Shimizu, sp. nov.), seven are new records from Japan (E. jilinensis Tang, 1990, E. laqueatus (Enderlein, 1921), E. multidens Chiu, 1954, stat. rev., E. puncticulatus Tang, 1990, E. stenophleps Cushman, 1937, E. vestigator (Smith, 1858), and E. zeugos Chiu, 1954, stat. rev.), 32 had already been recorded in Japan; three (E. biharensis Townes, Townes & Gupta, 1961, E. flavicaput (Morley, 1912), and E. merdarius (Gravenhorst, 1829)) have been erroneously recorded from Japan based on misidentifications, and four names that were previously on the Japanese list are deleted through synonymy. The following taxonomic changes are proposed: E. vacuus Gauld & Mitchell, 1981, syn. nov. (= E. formosensis (Uchida, 1928)); E. multidens stat. rev.; E. striatus Cameron, 1899, syn. nov. = E. lineolatus (Roman, 1913), syn. nov. = E. uniformis Chiu, 1954, syn. nov. = E. flatus Chiu, 1954, syn. nov. = E. gussakovskii Viktorov, 1957, syn. nov. = E. striolatus Townes, Townes & Gupta, 1961, syn. nov. = E. unicornis Rao & Nikam, 1969, syn. nov. = E. unicornis Rao & Nikam, 1970, syn. nov. (= E. pungens (Smith, 1874)); E. iracundus Chiu, 1954, syn. nov. (= E. sakaguchii (Matsumura & Uchida, 1926)); E. sigmatoides Chiu, 1954, syn. nov. (= E. shikokuensis (Uchida, 1928)); E. yamanakai (Uchida, 1930), syn. nov. (= E. shinkanus (Uchida, 1928)); E. ranunculus Chiu, 1954, syn. nov. (= E. yezoensis (Uchida, 1928)); and E. zeugos stat. rev. = E. henrytownesi Chao & Tang, 1991, syn. nov. In addition, the following new regional and country records are also provided: E. flavocephalus (Kirby, 1900), E. puncticulatus, and E. vestigator from the Eastern Palaearctic region, E. laqueatus from the Eastern Palaearctic and Oceanic regions, and E. maruyamanus (Uchida, 1928) from the Oriental region; E. abdominalis (Szépligeti, 1906) from Nepal, E. flavocephalus from Laos, E. formosensis from Laos and Malaysia, E. insinuator (Smith, 1860) from Taiwan, E. maruyamanus from India and Philippines, E. nigronotatus Cameron, 1903, E. riukiuensis (Matsumura & Uchida, 1926), and E. sakaguchii from Indonesia, E. pungens from 14 countries (Australia, Bhutan, Brunei, Indonesia, Laos, Malaysia, Nepal, New Caledonia, Papua New Guinea, Philippines, Solomon Islands, Sri Lanka, Tajikistan, and Taiwan), and E. yezoensis from South Korea. An identification key to all Japanese species of Enicospilus is proposed. Although 47 species are recognised in the present study, approximately 55 species could potentially be found in Japan based on ACE and Chao 1 estimators. The latitudinal diversity gradient of Enicospilus species richness is also tested in the Japanese archipelago based on the constructed robust taxonomic framework and extensive samples. Enicospilus species richness significantly increases towards the south, contrary to the 'anomalous' pattern of some other ichneumonid subfamilies.

The Roles of ROS Generation in RANKL-Induced Osteoclastogenesis: Suppressive Effects of Febuxostat.

Receptor activator of NF-κB ligand (RANKL), a critical mediator of osteoclastogenesis, is upregulated in multiple myeloma (MM). The xanthine oxidase inhibitor febuxostat, clinically used for prevention of tumor lysis syndrome, has been demonstrated to effectively inhibit not only the generation of uric acid but also the formation of reactive oxygen species (ROS). ROS has been demonstrated to mediate RANKL-mediated osteoclastogenesis. In the present study, we therefore explored the role of cancer-treatment-induced ROS in RANKL-mediated osteoclastogenesis and the suppressive effects of febuxostat on ROS generation and osteoclastogenesis. RANKL dose-dependently induced ROS production in RAW264.7 preosteoclastic cells; however, febuxostat inhibited the RANKL-induced ROS production and osteoclast (OC) formation. Interestingly, doxorubicin (Dox) further enhanced RANKL-induced osteoclastogenesis through upregulation of ROS production, which was mostly abolished by addition of febuxostat. Febuxostat also inhibited osteoclastogenesis enhanced in cocultures of bone marrow cells with MM cells. Importantly, febuxostat rather suppressed MM cell viability and did not compromise Dox's anti-MM activity. In addition, febuxostat was able to alleviate pathological osteoclastic activity and bone loss in ovariectomized mice. Collectively, these results suggest that excessive ROS production by aberrant RANKL overexpression and/or anticancer treatment disadvantageously impacts bone, and that febuxostat can prevent the ROS-mediated osteoclastic bone damage.

A new synonym for Enicospilus grammospilus (Enderlein, 1921) (Hymenoptera: Ichneumonidae: Ophioninae).

The megadiverse genus Enicospilus Stephens, 1835 (Hymenoptera: Ichneumonidae: Ophioninae) comprises over 700 valid species worldwide (e.g. Gauld Mitchell, 1981; Broad Shaw, 2016; Shimizu, 2017). Species of this genus are koinobiont endoparasitoids of moderately large sized lepidopteran larva (e.g. Gauld, 1985b; Broad Shaw, 2016). Members of this genus usually have "ophionoid facies", characterized by an orange-brown body, extremely large ocelli, long antennae, etc. (Gauld Huddleston, 1976). Ophionoid facies wasps including Enicospilus are attracted to light and considered to be crepuscular or nocturnal (e.g. Short et al., 2006; Shimizu Maeto, 2016; Shimizu Lima, 2018). These wasps usually exhibit a very limited number of diagnostic characters and as a consequence the delimitation and recognition of species is very difficult, and causes many taxonomic confusions including synonyms, homonyms and misidentifications.

A new species and a key to world species of the flavipes species-group of the genus Cotesia Cameron, 1891 (Hymenoptera: Braconidae: Microgastrinae) from Japan.

A new species of microgastrine parasitoid wasp (Hymenoptera: Braconidae), Cotesia testacea Fujie, Shimizu Fernandez-Triana sp. nov., is described from Japan and Korea. It belongs to the flavipes species-group, which now comprises seven described species, most of them economically important as biocontrol agents of cereal and sugarcane stem borer pests worldwide. The new species, currently known from marsh habitats in the Eastern Palaearctic, is morphologically similar and probably related to the Western Palaearctic Cotesia ferruginea. A key to world species of the flavipes species-group is also provided.

Recognition of the genus Habrocampulum Gauld, 1976 (Hymenoptera: Ichneumonidae: Anomaloninae) from Japan, with a new combination and a key to the species.

Recognition of the genus Habrocampulum Gauld, 1976 (Hymenoptera:Ichneumonidae: Anomaloninae) from Japan, with a new combination and a key to the species.

Revision of the Taiwanese species of the genus Leptophion Cameron, 1901 (Hymenoptera: Ichneumonidae: Ophioninae), with a discussion of their phenology and distribution.

We revise the Taiwanese species of the ophionine genus Leptophion Cameron, 1901. As a result, three species, including a new species, are recognized. Two of them were identified as L. maculipennis (Cameron, 1905) and L. radiatus (Uchida, 1956), both of which had been previously recorded in Taiwan; we redescribe them based on Taiwanese specimens. We describe a single new species as L. giganteus Shimizu & Watanabe, sp. nov. The phenology and distribution of the species are briefly discussed. A key to the Taiwanese species of Leptophion and additional couplets to the key proposed by Gauld & Mitchell (1981) are also provided.

Discovery of the genus Leptophion Cameron, 1901, from Japan and the Palaearctic region, with description of two new species (Hymenoptera: Ichneumonidae: Ophioninae).

The genus Leptophion Cameron, 1901, which belongs to the subfamily Ophioninae Shuckard, 1840, is recorded from Japan and the Palaearctic region for the first time based on two new species, L. parvus sp. nov. from Amamioshima Island and L. septentrionis sp. nov. from Yakushima and Amamioshima Island. A key to the Japanese and Taiwanese species is also provided.

Application of chaotic dynamics in a recurrent neural network to control: hardware implementation into a novel autonomous roving robot.

Originating from a viewpoint that complex/chaotic dynamics would play an important role in biological system including brains, chaotic dynamics introduced in a recurrent neural network was applied to control. The results of computer experiment was successfully implemented into a novel autonomous roving robot, which can only catch rough target information with uncertainty by a few sensors. It was employed to solve practical two-dimensional mazes using adaptive neural dynamics generated by the recurrent neural network in which four prototype simple motions are embedded. Adaptive switching of a system parameter in the neural network results in stationary motion or chaotic motion depending on dynamical situations. The results of hardware implementation and practical experiment using it show that, in given two-dimensional mazes, the robot can successfully avoid obstacles and reach the target. Therefore, we believe that chaotic dynamics has novel potential capability in controlling, and could be utilized to practical engineering application.