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The trictenotomid genus Autocrates Thomson, 1860 is remarkable for its large and robust adults, but its larval morphology and bionomics have been unknown over the last 160 years. Here, we describe and illustrate in detail the eggs, and also the first and last instar larva and the pupa of Autocrates maqueti Drumont, 2006, based on specimens reared from identified adults collected in South Korea. The first instar larva is very similar to the known trictenotomid larvae of the genus Trictenotoma Gray, 1832, sharing the following features: distinctly flattened and parallel-sided, well-sclerotized head and largely unpigmented postcephalic body, lyre-shaped frontal arms, very short coronal suture, five pairs of stemmata, asymmetrical and tridentate mandibles with mola, simple and apically upturned urogomphi, and absence of longitudinal ridges on the thorax and abdomen. Differences occur in the primary chaetotaxy. The last instar larva is almost identical with Trictenotoma in general appearance, but the longitudinal tergal ridges of Autocrates are much denser and more widely distributed than those of Trictenotoma. Our preliminary assessment of features of immature stages confirms a close relationship of Trictenotomidae with the “salpingid group”, i.e. Salpingidae, Boridae, Pyrochroidae and Pythidae. A sister group relationship with Pythidae is likely. The putative synapomorphy is the subdivision of the ventral element of tergites IX, even though this condition is not visible in the first instars of Autocrates, and quite indistinct (but recognizable) in the last larval stage. The complete mitochondrial genome of A. maqueti is provided. A preliminary phylogenetic analysis of trictenotomid species is presented using two mitochondrial genes (16S and COI). The morphology of eggs and biological information on feeding and oviposition behaviors are also provided with photographs of adults and eggs.

Freshwater leeches belonging to the genus Glossiphonia Johnson, 1816 are fairly common benthic annelids in freshwater ecosystems throughout Eurasia, but the taxonomy of this group remains poorly resolved, and the species content of some local faunas is unsatisfactorily known. For example, it was thought that the only widespread species, Glossiphonia complanata (Linnaeus, 1758), dwells in Northeast Asia. Here, we revise the Northeast Asian Glossiphonia leeches with an integrative taxonomic approach. This revision is based on the largest DNA-sequence and morphological datasets collected to date. Two Glossiphonia species are recorded from the region. First, Glossiphonia koreaensissp. nov., a putative regional endemic species, was discovered in South Korea. Second, a valid name and complete description are provided for Glossiphonia mollissima Moore, 1898 (non-Grube, 1871). The latter name was wrongly applied through misidentification and, according to ICZN (Article 49), is unavailable. Hence, we propose a new name, Glossiphonia mooreisp. nov., in memory of Dr. J. Percy Moore, who discovered this leech. It is a prospective trans-Beringian species, ranging through Eastern Siberia, the Russian Far East, and Alaska (although Alaskan records are based on nonsequenced samples), showing a significant proportion of melanic individuals in samples north of the Arctic Circle. The latter pattern may reflect substrate-induced cryptic coloration (camouflage), but this hypothesis needs to be statistically checked in the future. Finally, a complete checklist of Glossiphonia species is presented.

Inflammation causes a protective immune response, which can be observed by examining the inflammatory responses of macrophages. Macrophages release various immunostimulatory factors when destroying external pathogens. We induced lipopolysaccharides (LPS) in RAW 264.7 cells, a macrophage cell line, to determine whether Helixor-M can cause immuno-suppression. Helixor-M is known to have anticancer and immune effects. However, an indicator that regulates immunity has not been clearly confirmed. To this end, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was conducted to confirm Helixor-M was not cytotoxic. Western blotting and real-time polymerase chain reaction (RT-PCR) confirmed the anti-inflammatory effects. Additionally, immunofluorescence assay confirmed the translocation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65, a representative inflammatory pathway. Helixor-M was found to be non-cytotoxic, induce the NF-κB pathway, and reduce the levels of pro-inflammatory cytokine and mitogen-activated protein kinase (MAPK). We found Helixor-M affected the PI3K/AKT/JNK pathway. Therefore, we confirmed Helixor-M acts as an anti-inflammatory agent through NF-κB, TLR4 and PI3K inhibition and that it could be an effective immunosuppressive drug.

Background aims Tracking administered natural killer (NK) cells in vivo is critical for developing an effective NK cell-based immunotherapy against human hepatocellular carcinoma (HCC). Here the authors established a new molecular imaging using ex vivo-activated NK cells and investigated real-time biodistribution of administered NK cells during HCC progression. Methods Ex vivo-expanded NK cells from healthy donors were labeled with a near-infrared lipophilic cytoplasmic dye, and their proliferation, surface receptor expression and cytotoxicity activity were evaluated. Human HCC HepG2 cells were implanted into the livers of NOD.Cg-Prkdcscid IL2rgtm1Wjl/SzJ (NSG) mice. The authors administered 1,1’-dioctadecyltetramethyl indotricarbocyanine iodide (DiR)-labeled NK cells intravenously to non-tumor-bearing and intrahepatic HCC tumor-bearing NSG mice. Fluorescent imaging was performed using a fluorescence-labeled organism bioimaging instrument. Single cell suspensions from the resected organs were analyzed using flow cytometry. Results The fluorescent DiR dye was nontoxic and did not affect the proliferation or surface receptor expression levels of the NK cells, even at high doses. The administered DiR-labeled NK cells immediately migrated to the lungs of the non-tumor-bearing NSG mice, with increased NK cell signals evident in the liver and spleen after 4 h. NK cells migrated to the intrahepatic tumor-bearing livers of both early- and late-stage HCC mice within 1 h of injection. In early-stage intrahepatic tumor-bearing mice, the fluorescence signal increased in the liver until 48 h post-injection and decreased 7 days after NK injection. In late-stage HCC, the NK cell fluorescence signal was the highest in the liver for 7 days after NK injection and persisted for 14 days. The purity of long-term persistent CD45+CD56+CD3− NK cells was highest in early- and late-stage HepG2-bearing liver compared with normal liver 2 weeks after NK injection, whereas highest purity was still observed in the lungs of non-tumor-bearing mice. In addition, Ki-67 expression was detected in migrated human NK cells in the liver and lung up to 72 h after administration. With HepG2 tumor progression, NK cells reduced the expression of NKp30 and NKG2D. Conclusions Administered NK cells were successfully tracked in vivo by labeling the NK cells with near-infrared DiR dye. Highly expanded, activated NK cells migrated rapidly to the tumor-bearing liver, where they persisted for 14 days after administration, with high purity of CD45+CD56+CD3− NK cells. Liver biodistribution and persistence of administered NK cells showed significantly different accumulation patterns during HCC progression.

Currently, the validity of many nominal bivalve species of the genus Corbicula endemic to the Russian Far East and South Korea needs a critical reassessment. In this study, we clarify the taxonomic status of Corbicula species of this area based on a combination of molecular genetic, conchological and anatomical data. According to our results, four Corbicula lineages, corresponding to the nominal species Corbicula japonica, Corbicula elatior, Corbicula leana and Corbicula fluminea, can be delineated in samples collected in the Primorye and Khabarovsk regions of Russia and South Korea. Two species endemic to the Russian Far East (i.e. Corbicula finitima and Corbicula lindholmi) are considered here as junior synonyms of the species C. japonica, which is widely distributed in estuarine habitats around the Japanese Archipelago, Sakhalin Island, southern Kurile Islands, Primorye and Khabarovsk regions, Korean Peninsula and China. Three nominal species described from the Lower Amur basin (Corbicula amurensis, Corbicula nevelskoyi and Corbicula sirotskii) appeared to be synonyms of C. elatior, whose range covers the Korean Peninsula, Primorye and Khabarovsk regions and, perhaps, China. We delineated several colour morphs of C. fluminea and C. japonica. The distinctness between these colour morphs can be attributed to both heritable and environmental factors.

The cover image is based on the Case Report A case of leukaemia cutis in a dog with T‐cell chronic lymphocytic leukaemia by Hyeona Bae et al., https://doi.org/10.1002/vms3.749. [Image: see text]

Canine natural killer (NK) cells are large, granular lymphocytes that are neither B lymphocytes nor T lymphocytes. However, it has been reported that canine NK cells share some of the phenotypic characteristics of T lymphocytes, such as CD3 and CD5. Studies are needed to assess the safety of canine NK cells for immunotherapy, especially because the safety of using allogeneic NK cells as an immunotherapy for dogs has yet to be shown. In this study, the safety of cultured canine NK cells was assessed using a xenogeneic mouse model of graft-versus-host disease (GVHD). Mice were injected with either canine peripheral blood mononuclear cells (PBMCs) or cultured NK cells for 2 or 3 weeks. Data were then collected on changes in mice body weights, disease severity scores, and survival rates. Histopathological and immunohistochemical evaluations were also performed. All mice injected with canine PBMCs died within 45 days after injection. Severe clinical signs were caused by GVHD. The histopathological and immunohistochemical evaluations showed that mice injected with canine PBMCs had multiple lesions, including necrosis in their lungs, livers, kidneys, and stomachs, and the injected cells were present around the lesions. By contrast, no mice injected with cultured NK cells without removing the CD3+TCR– cells exhibited any clinical abnormalities. Moreover, they all survived the 90-day experimental period without exhibiting any histopathological changes. Accordingly, the results of this study suggest that canine NK cells do not cause significant side effects such as GVHD and allogeneic NK cells can safely be used for cancer immunotherapy in dogs.

The leaf beetle genus Suinzona, consisting of over 20 species, is mainly known for species from Southwest China, and its closely related genus Potaninia, with only two species, is widely distributed in South China and upper Southeast Asia. Despite recent morphological studies, the status of these taxa has long been controversial. Here, we investigated the taxonomic status and phylogenetic relationships of the genera Suinzona and Potaninia within Chrysomelinae using molecular, morphological and ecological data. Molecular phylogenetic analysis supported that they should be regarded as distinct genera, which is consistent with morphological evidence, e.g., well-developed/reduced hind wings. Based on combined evidence from examination of larval and adult morphology, host plants and mitochondrial genomes, we demonstrate that P. cyrtonoides should be placed in the genus Suinzona and that specimens from South Korea represent a new species. Suinzona borowieci sp. nov., occurring in narrow strips of habitat, shows high levels of genetic divergence and distantly related host plants between populations. The population differentiation seems to be correlated with its non-functional wings causing reduced dispersal ability and genetic isolation. Several populations have declined dramatically over the last few decades due to loss of habitat and thus are in need of protection as conservation units.

Aphyocypris chinensis Gunther, 1868 is a small freshwater fish of the family Xenocyprididae (Cypriniformes). In this study, we determined its complete mitochondrial genome and phylogenetic position in Cypriniformes. The complete mitochondrial genome is 16,608 bp in size, containing 13 protein-coding genes, two RNAs, 22 tRNAs, and a control region. It has the typical vertebrate mitochondrial gene arrangement. Our mitogenomic phylogeny revealed that A. chinensis belongs to Xenocyprididae, rather than Danionidae. This mitogenome information could play an essential role in resolving the conflict over its current taxonomic status in Cypriniformes.

Apigenin, an aromatic compound, exhibits antioxidant, anti‑inflammatory and anti‑viral effects. The present study aimed to investigate the effects of apigenin on cell proliferation and apoptosis of human melanoma cells A375P and A375SM. Therefore, melanoma cells were treated with apigenin to determine its anti‑proliferative and survival effects, using wound healing and MTT assays. The results revealed that melanoma cell viability was decreased in a dose‑dependent manner. Furthermore, chromatin condensation, indicating apoptosis, was significantly increased in a dose‑dependent manner, as demonstrated by DAPI staining. In addition, increased apoptosis rate following treatment with apigenin was confirmed by Annexin V‑propidium iodide staining. The changes in the expression levels of apoptosis‑related proteins in A375P and A375SM melanoma cells were subsequently detected using western blot analysis. The results demonstrated that the protein expression levels of Bcl‑2 were decreased, whereas those of Bax, cleaved poly ADP‑ribose polymerase, cleaved caspase‑9 and p53 were upregulated in a dose‑dependent manner in apigenin‑treated cells compared with those noted in untreated cells. In addition, in apigenin‑treated A375P cells, phosphorylated (p)‑p38 was upregulated and p‑extracellular signal‑regulated kinase (ERK), p‑c‑Jun N‑terminal kinase (JNK) and p‑protein kinase B (Akt) were downregulated. However, in A375SM cells, apigenin treatment increased p‑ERK and p‑JNK and decreased p‑p38 and p‑Akt protein expression levels. Subsequently, the inhibitory effect of apigenin on tumor growth was investigated in vivo. Tumor volume was significantly reduced in the 25 and 50 mg/kg apigenin‑treated groups compared with the control group. Additionally, a TUNEL assay was performed to detect apoptotic cells. Immunohistochemical staining also revealed elevated p‑ERK expression in the apigenin‑treated group compared with the control group. Overall, the findings of the present study indicated that apigenin attenuated the growth of A375SM melanoma cells by inducing apoptosis via regulating the Akt and mitogen‑activated protein kinase signaling pathways.

Purpose We investigated whether adoptive cell therapy with ex vivo–activated natural killer (NK) cells enhances the therapeutic efficacy of local tumor radiation therapy (RT) using a human triple-negative breast cancer xenograft model. Methods and Materials NK cells from healthy donors were expanded ex vivo. MDA-MB-231/Luc-GFP cells were subcutaneously implanted into the thighs of NSG mice. The animals were divided into 4 experimental groups: control, RT, NK, and RT + NK. On day 17 after tumor implantation, tumors from the RT groups were irradiated. The ex vivo–expanded NK cells were intravenously administered twice, on days 17 and 19. Primary and secondary tumors were evaluated using long-term bioluminescence imaging, and histopathology was performed on resected tumor tissue specimens. Results The luciferase signals of the primary tumors in the RT + NK group were significantly lower than those of comparably sized primary tumors in the RT group. The long-term migration and infiltration of NK cells into the primary tumor sites were significantly higher in RT + NK than in NK mice. Moreover, lymphatic metastasis to the axillary lymph nodes and liver and lung metastases were highly suppressed in the RT + NK group, as demonstrated by BLI and p53 immunohistochemistry. The long-term survival of the RT + NK group was significantly higher than that of the RT or NK groups. Conclusions Reduction in tumor burden by combining RT and systemic NK cell therapy improved the suppression of primary tumor growth, with efficient NK cell migration and penetration into the primary tumor site. Administered NK cells were maintained in the primary tissue for a significantly longer time in RT + NK group compared with NK group. Both lymphatic spread and distant metastasis to the lungs and liver were effectively suppressed by the combined therapy.

Myricetin, a flavonoid found in fruits and vegetables, is known to have antioxidant and anticancer effects. However, the anticancer effects of myricetin on SK‑BR‑3 human breast cancer cells have not been elucidated. In the present study, the anticancer effects of myricetin were confirmed in human breast cancer SK‑BR‑3 cells. As the concentration of myricetin increased, the cell viability decreased. DAPI (4',6‑diamidino‑2‑phenylindole) and Annexin V/PI staining also revealed a significant increase in apoptotic bodies and apoptosis. Western blot analysis was performed to confirm the myricetin‑induced expression of apoptosis‑related proteins. The levels of cleaved PARP and Bax proteins were increased, and that of Bcl‑2 was decreased. The levels of proteins in the mitogen‑activated protein kinase (MAPK) pathway were examined to confirm the mechanism of myricetin‑induced apoptosis, and it was found that the expression levels of phosphorylated c‑Jun N‑terminal kinase (p‑JNK) and phosphorylated mitogen‑activated protein kinases (p‑p38) were increased, whereas that of phosphorylated extracellular‑regulated kinase (p‑ERK) was decreased. It was also demonstrated that myricetin induced autophagy by promoting autophagy‑related proteins such as microtubule‑associated protein 1A/1B‑light chain 3 (LC 3) and beclin 1. In addition, 3‑methyladenine (3‑MA) was used to evaluate the association between cell viability and autophagy in cells treated with myricetin. The results showed that simultaneous treatment with 3‑MA and myricetin promoted the apoptosis of breast cancer cells. Furthermore, treatment with a JNK inhibitor reduced cell viability, promoted Bax expression, and reduced the expression of p‑JNK, Bcl‑2, and LC 3‑II/I. These results suggest that myricetin induces apoptosis via the MAPK pathway and regulates JNK‑mediated autophagy in SK‑BR‑3 cells. In conclusion, myricetin shows potential as a natural anticancer agent in SK‑BR‑3 cells.

Aphyocypris chinensis Günther, 1868 is a small freshwater fish of the family Xenocyprididae (Cypriniformes). In this study, we determined its complete mitochondrial genome and phylogenetic position in Cypriniformes. The complete mitochondrial genome is 16,608 bp in size, containing 13 protein-coding genes, two RNAs, 22 tRNAs, and a control region. It has the typical vertebrate mitochondrial gene arrangement. Our mitogenomic phylogeny revealed that A. chinensis belongs to Xenocyprididae, rather than Danionidae. This mitogenome information could play an essential role in resolving the conflict over its current taxonomic status in Cypriniformes.

In multiple myeloma (MM), the impaired function of several types of immune cells favors the tumor's escape from immune surveillance and, therefore, its growth and survival. Tremendous improvements have been made in the treatment of MM over the past decade but cellular immunotherapy using dendritic cells, natural killer cells, and genetically engineered T-cells represent a new therapeutic era. The application of these treatments is growing rapidly, based on their capacity to eradicate MM. In this review, we summarize recent progress in cellular immunotherapy for MM and its future prospects.

This study aimed to identify potential biomarkers of canine pyometra and their correlations with clinical parameters. First, 90 dogs with pyometra and 26 healthy female dogs were compared. Then, paired samples (before and after ovariohysterectomy) from 22 dogs with pyometra and 9 healthy controls from the initial cohort were compared. Concentrations of acute inflammatory proteins, C-reactive protein (CRP) and serum amyloid A (SAA), and cell-free DNA (cfDNA), were significantly higher in dogs with pyometra than in clinically healthy dogs. Cell-free DNA was the most sensitive biomarker for systemic inflammation, based on the receiver operating characteristic curve analysis (area under the curve = 0.959). In addition, cfDNA and CRP were significantly associated with inflammation and organ injury-related clinical parameters. Following the surgical removal of the inflamed uterus, interleukin-6 (IL-6), high-mobility group box 1 (HMGB1), and procalcitonin (PCT) significantly decreased, whereas changes in CRP, SAA, and cfDNA were not significant. These findings indicate that cfDNA, CRP, and SAA are potential clinical biomarkers of systemic inflammation in dogs with pyometra and PCT, IL-6, and HMGB1 are potential biomarkers of clinical recovery.Cette étude visait à identifier les biomarqueurs potentiels du pyomètre canin et leurs corrélations avec les paramètres cliniques. Tout d’abord, 90 chiens avec pyomètre et 26 chiennes en bonne santé ont été comparés. Ensuite, des échantillons appariés (avant et après ovariohystérectomie) de 22 chiens avec pyomètre et neuf témoins sains de la cohorte initiale, ont été comparés.Les concentrations des protéines inflammatoires aiguës, protéine C réactive (CRP) et amyloïde sérique A (SAA), et d’ADN acellulaire (cfDNA), étaient significativement plus élevées chez les chiens atteints de pyomètre que chez les chiens cliniquement sains. L’ADN acellulaire était le biomarqueur le plus sensible pour l’inflammation systémique, sur la base de l’analyse de la courbe caractéristique de fonctionnement du récepteur (aire sous la courbe = 0,959). De plus, le cfDNA et la CRP étaient significativement associés à l’inflammation et aux paramètres cliniques liés aux lésions aux organes.Après l’ablation chirurgicale de l’utérus enflammé, l’interleukine-6 (IL-6), la protéine HMGB1 (« high-mobility groupe box 1 ») et la procalcitonine (PCT) ont significativement diminué, alors que les changements de CRP, SAA et cfDNA n’étaient pas significatifs. Ces résultats indiquent que cfDNA, CRP et SAA sont des biomarqueurs cliniques potentiels de l’inflammation systémique chez les chiens avec pyomètre et PCT, IL-6 et HMGB1 sont des biomarqueurs potentiels de récupération clinique.(Traduit par Docteur Serge Messier).

Interleukin-15 (IL-15) is a pleiotropic cytokine that plays pivotal roles in innate and adaptive immunity. It is also a promising cytokine for treating cancer. Despite growing interest in its use as an immunotherapeutic, its safety and immunological effects in dogs have not been reported. In this study, healthy dogs were given recombinant canine IL-15 (rcIL-15) intravenously at a daily dose of 20 μg/kg for 8 days and monitored for 32 days to determine the safety and immunological effects of rcIL-15. The repeated administration of rcIL-15 was well tolerated, did not cause any serious side effects, and promoted the selective proliferation and activation of canine anti-cancer effector cells, including CD3+CD8+ cytotoxic T lymphocytes, CD3+CD5dimCD21–, and non-B/non-T NK cell populations, without stimulating Treg lymphocytes. The rcIL-15 injections also stimulated the expression of molecules and transcription factors associated with the activation and effector functions of NK cells, including CD16, NKG2D, NKp30, NKp44, NKp46, perforin, granzyme B, Ly49, T-bet, and Eomes. These results suggest that rcIL-15 might be a valuable therapeutic adjuvant to improve immunity against cancer in dogs.

We determined the complete mitochondrial genome sequence of Squalidus chankanensis tsuchigae (Cypriniformes: Cyprinidae). It is 16,603 bp long, with 13 protein-coding genes, 22 transfer RNAs, 2 ribosomal RNA genes, and 1 control region, which is 927 bp long, located between tRNApro and tRNAphe. The overall base composition is as follows: 29.98% A, 16.86% G, 25.44% T, and 27.72% C, with a slight AT bias. These results provide necessary data for phylogenetic studies on Squalidus species.

Light-emitting diodes (LED) based displays are highlighted because of their interesting features compared with others. Especially, the LED display with active matrix (AM) has additional advantages such as low power consumption, elimination of current cross talk, and so on. However, AM- driving LED displays are fabricated on active substrates with transistors so that there are many problems like high-cost, difficulty in enhancing yield, and repair problem. As an alternative, an AM-driving LED display on a passive substrate has been developed in this study. One pixel is composed of RGB mini-LEDs and a control IC, fabricated using the CMOS technology. It can decrease the cost of a substrate compared with other AM LED displays, and keep constant the complexity of substrate regardless of the variations of the pitch of mini-LEDs. To mount mini-LEDs and control ICs at the same time needs the technical innovations because they have different surface finishes. Moreover, there is no solder resist layer on a substrate because of the fine-pitch of mini-LEDs and control ICs in a single pixel. The Solder-on-Pad (SoP), fluxing underfill, and Laser-Assisted Bonding (LAB) technologies are developed to implement digital signage modules, successfully.

In a large-scale IoT environment such as a future smart city, various sensors and autonomous cars/drones/robots collect and process a wide variety of data. However existing monolithic or silo style operation of smart city makes hard to solve the problem when an unseen complex event occurs. In this paper, we suggest a structure and method of distributed intelligence processing which mimics the function and network structure of the brain in order to efficiently control the complex situations. Our approach utilizes local and rapid processing in parallel as well as global and exploratory processing through cooperation and competition between local intelligence. It will give us an opportunity to realize the autonomous problem solving on a large scale hyper-connected network infrastructure, by finding an emergence structure of distributed intelligence that mimics human consciousness process.

Oxidative stress plays a vital role in neurodegenerative diseases. Cornus officinalis (CC) has a wide range of pharmacological activities (e.g., antioxidant, neuroprotective, and anti-inflammatory). The present study was undertaken to elucidate the neuroprotective mechanism of CC and fermented CC (FCC) on stress and H2O2-induced oxidative stress damage in rats and SH-SY5Y cells. A dose of 100 mg/kg CC or FCC was orally administered to rats 1 h prior to immobilization 2 h per day for 14 days. CC, especially FCC administration decreased immobility time in forced swim test (FST), effectively alleviated the oxidative stress, and remarkably decreased corticosterone, &beta, endorphin and increased serotonin levels, respectively. In cells, CC and FCC significantly inhibited reactive oxygen species (ROS) generation, lactate dehydrogenase (LDH) release and significantly increased the genes expression of antioxidant and neuronal markers, such as superoxide dismutase (SOD), catalase (CAT), and brain-derived neurotrophic factor (BDNF). Moreover, the pro-apoptotic factor Bax and anti-apoptotic factor Bcl-2 (Bax/Bcl-2) ratio was regulated by CC and FCC pretreatment. Both in rats and cells, CC and FCC downregulated mitogen-activated protein kinase (MAPK) phosphorylation. Taken together, these results demonstrated that CC and particularly FCC ameliorated oxidative stress and may be used on the neuroprotection.