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Activation of Gq protein-coupled receptors triggers the phospholipase C (PLC) pathway, which yields a pair of second messengers: diacylglycerol (DG) and inositol 1,4,5-trisphosphate (IP

We describe the current state of speciation in the species complex of the marine gastropod Patelloida saccharina, using mitochondrial COI, 16S RNA, nuclear histone 3 gene and single-nucleotide polymorphism (SNP) markers from 165 specimens collected from 37 localities across the Japanese archipelago. In total, 28 unique COI haplotypes, 16 16S haplotypes, 11 histone 3 haplotypes and 148 734 SNPs were obtained. Genetic assignments and phylogenetic trees recovered three genetically distinct lineages: P. saccharina saccharina, P. saccharina lanx and an unknown P. sp. from Shionomisaki, Wakayama Prefecture. The divergence between P. saccharina saccharina and P. saccharina lanx was estimated to have occurred around 44 000 years ago—too recent to have accumulated the morphological differences that have confounded taxonomic identification. Continuous gene exchange after an initial split is suggested based on demographic history analyses, which allowed for introgression of the P. saccharina lanx genome and the proliferation of intermediate individuals. Speciation, despite the existence of a contact zone in the Ryukyu Islands, possibly occurred due to the eventual reproductive isolation that followed the initial split with gene flow. Patelloida saccharina saccharina and P. saccharina lanx were strongly supported by our results as two separate species, and a revision of the current taxonomic descriptions based on their phylogeny, demographic history and species distribution is proposed.

PDF file - 63K, Differential PLA2G7 expression in colorectal adenocarcinomas versus normal tissues, as determined by immunohistochemical analyses reported in the literature.

PDF file - 92K, Supplementary methodology, including materials utilized, siRNA sequences, a description of immunoblot and immunohistochemical procedures, and statistical considerations.

PDF file - 122K, Protein phosphatases PP1 and PP2 do not rescue cPAF-induced de-phosphorylation of pAkt473

PDF file - 144K, PKC inhibition does not affect cPAF-induced de-phosphorylation of pAkt473

PDF file - 60K, cPAF-induced de-phosphorylation of pAKT473 does not require Gαi/ Gαo or PKA

Purpose Although the usefulness of O6-methylguanine DNA methyltransferase (MGMT) promoter methylation analysis for predicting response to chemoradiotherapy and the prognosis of patients with glioblastoma has been widely reported, there is still no consensus regarding how to define MGMT promoter methylation percentage (MGMTpm%) cutoffs by pyrosequencing method. The aim of this study was to determine the optimal cutoff value of MGMT promoter methylation status using volumetric analysis focused on the tumor volume ratio (TVR) measured by MRI. Methods This retrospective study included newly diagnosed IDH wild-type glioblastoma patients with residual tumor after surgery, followed by local radiotherapy with temozolomide. TVR was defined as the tumor volume at 6 months after the initial chemoradiotherapy administration divided by the tumor volume before the start of therapy. The mean MGMTpm% of 16 CpG islands (74–89) was analyzed using pyrosequencing. We statistically analyzed the correlation between MGMTpm%, TVR, and change in Karnofsky performance status. Results The study included 44 patients with residual tumors. Thirteen (92.9%) of 14 patients with MGMTpm% ≥ 23.9% showed 50% or more volumetric response, leading to prolonged survival, and 17 (70.8%) of 24 patients with MGMTpm% Conclusion Evaluation of MGMTpm% by pyrosequencing is important in predicting the volumetric response and prognosis of glioblastoma patients with residual tumors.

Brittle stars (class Ophiuroidea) are marine invertebrates comprising approximately 2, 100 extant species, and are considered to constitute the most diverse taxon of the phylum Echinodermata. As a non-invasive method for monitoring biodiversity, we developed two new sets of PCR primers for metabarcoding environmental DNA (eDNA) from brittle stars. The new primer sets were designed to amplify 2 short regions of the mitochondrial 16S rRNA gene, comprising a conserved region (111–115 bp, 112 bp on average; named “16SOph1”) and a hyper-variable region (180–195 bp, 185 bp on average; named “16SOph2”) displaying interspecific variation. The performance of the primers was tested using eDNA obtained from two sources: a) rearing water of an 2.5 or 170 L aquarium tanks containing 15 brittle star species and b) from natural seawater collected around Misaki, the Pacific coast of central Japan, at depths ranging from shallow (2 m) to deep (> 200 m) sea. To build a reference library, we obtained 16S rRNA sequences of brittle star specimens collected from around Misaki and from similar depths in Japan, and sequences registered in International Nucleotide Sequence Database Collaboration. As a result of comparison of the obtained eDNA sequences with the reference library 37 (including cryptic species) and 26 brittle star species were detected with certain identities by 16SOph1 and 16SOph2 analyses, respectively. In shallow water, the number of species and reads other than the brittle stars detected with 16SOph1 was less than 10% of the total number. On the other hand, the number of brittle star species and reads detected with 16SOph2 was less than half of the total number, and the number of detected non-brittle star metazoan species ranged from 20 to 46 species across 6 to 8 phyla (only the reads at the “Tank” were less than 0.001%). The number of non-brittle star species and reads at 80 m was less than 10% with both of the primer sets. These findings suggest that 16SOph1 is specific to the brittle star and 16SOph2 is suitable for a variety of marine metazoans. It appears, however, that further optimization of primer sequences would still be necessary to avoid possible PCR dropouts from eDNA extracts. Moreover, a detailed elucidation of the brittle star fauna in the examined area, and the accurate identification of brittle star species in the current DNA databank is required., 汲んだ水から深海生物の種類を判別 --世界初「クモヒトデメタバーコーティング」技術を開発--. 京都大学プレスリリース.

Meningioma is the most common intracranial tumor, with generally favorable patient prognosis. However, patients with malignant meningioma typically experience recurrence, undergo multiple surgical resections, and ultimately have a poor prognosis. Thus far, effective chemotherapy for malignant meningiomas has not been established. We recently reported the efficacy of eribulin (Halaven ®) for glioblastoma with a telomerase reverse transcriptase (TERT) promoter mutation. This study investigated the anti-tumor effect of eribulin against TERT promoter mutation-harboring human malignant meningioma cell lines in vitro and in vivo. Two meningioma cell lines, IOMM-Lee and HKBMM, were used in this study. The strong inhibition of cell proliferation by eribulin via cell cycle arrest was demonstrated through viability assay and flow cytometry. Apoptotic cell death in malignant meningioma cell lines was determined through vital dye assay and immunoblotting. Moreover, a wound healing assay revealed the suppression of tumor cell migration after eribulin exposure. Intraperitoneal administration of eribulin significantly prolonged the survival of orthotopic xenograft mouse models of both malignant meningioma cell lines implanted in the subdural space (p < 0.0001). Immunohistochemistry confirmed apoptosis in brain tumor tissue treated with eribulin. Overall, these results suggest that eribulin is a potential therapeutic agent for malignant meningiomas.

Glioblastomas (GBM) often acquire resistance against temozolomide (TMZ) after continuous treatment and recur as TMZ‐resistant GBM (TMZ‐R‐GBM). Lomustine (CCNU) and nimustine (ACNU), which were previously used as standard therapeutic agents against GBM before TMZ, have occasionally been used for the salvage therapy of TMZ‐R‐GBM; however, their efficacy has not yet been thoroughly examined. Therefore, we investigated the antitumor effects of CCNU and ACNU against TMZ‐R‐GBM. As a model of TMZ‐R‐GBM, TMZ resistant clones of human GBM cell lines (U87, U251MG, and U343MG) were established (TMZ‐R‐cells) by the culture of each GBM cells under continuous TMZ treatment, and the antitumor effects of TMZ, CCNU, or ACNU against these cells were analyzed in vitro and in vivo. As a result, although growth arrest and apoptosis were triggered in all TMZ‐R‐cells after the administration of each drug, the antitumor effects of TMZ against TMZ‐R‐cells were significantly reduced compared to those of parental cells, whereas CCNU and ACNU demonstrated efficient antitumor effects on TMZ‐R‐cells as well as parental cells. It was also demonstrated that TMZ resistance of TMZ‐R‐cells was regulated at the initiation of DNA damage response. Furthermore, survival in mice was significantly prolonged by systemic treatment with CCNU or ACNU but not TMZ after implantation of TMZ‐R‐cells. These findings suggest that CCNU or ACNU may serve as a therapeutic agent in salvage treatment against TMZ‐R‐GBM., We investigated the antitumor effects of lomustine (CCNU) and nimustine (ACNU), which were previously used as standard therapeutic agents for glioblastomas (GBM), against the model cells of human GBM cases, which gained acquired temozolomide (TMZ) resistance after continuous treatment by TMZ (TMZ‐R‐cells). We discovered that the antitumor effects of TMZ against TMZ‐R‐cells were significantly reduced compared to those of parental cells, whereas CCNU and ACNU demonstrated efficient antitumor effects on TMZ‐R‐cells as well as parental cells both in vitro and in vivo. In addition, it was also demonstrated that TMZ resistance of TMZ‐R‐cells was regulated at the level of DNA damage response initiation. These findings suggest that CCNU or ACNU may serve as a therapeutic agent in salvage treatment against GBM cases with acquired TMZ resistance.

The mammalian oviductal lumen is a specialized chamber that provides an environment that strictly regulates fertilization an early embryogenesis, the regulatory mechanisms to gametes/zygote are still largely unknown. In this report, we studied the oviductal regulation of early embryonic development usingOvgp1(a gene encoding an oviductal humoral factor, OVGP1)-knockout (KO) hamsters. The experimental results revealed the following: 1) FemaleOvgp1-KO hamsters fail to produce any litters at all; 2) In the oviducts from KO animal, fertilized eggs are sometimes identified, but their morphology shows abnormal features; 3) The number of implantations in the KO females is evidently low; 4) Even if implantations occur, the embryos develop abnormally and eventually become embryonic lethal; and 5)Ovgp1-KO females transferred to wild-type females produce KO egg-derived litters, but the reverse experiment does not. These results suggest that OVGP1-mediated physiological events are crucial for early embryonic developmentin vivo. This animal model shows that the fate of the fertilized egg is not only genetically determined, but that the surrounding oviductal microenvironment plays a pivotal role in normal embryonic development.Summary statementDeficiency an oviductal humoral factor (OVGP1) caused female infertility in the golden hamsters. The presence or absence of OVGP1 has significant physiological effects on early embryonic developmentin vivo.

The aim of the present study was to determine which individual or combined CpG sites among O6-methylguanine DNA methyltransferase CpG 74–89 in glioblastoma mainly affects the response to temozolomide resulting from CpG methylation using statistical analyses focused on the tumor volume ratio (TVR). We retrospectively examined 44 patients who had postoperative volumetrically measurable residual tumor tissue and received adjuvant temozolomide therapy for at least 6 months after initial chemoradiotherapy. TVR was defined as the tumor volume 6 months after the initial chemoradiotherapy divided by that before the start of chemoradiotherapy. Predictive values for TVR as a response to adjuvant therapy were compared among the averaged methylation percentages of individual or combined CpGs using the receiver operating characteristic curve. Our data revealed that combined CpG 78 and 79 showed a high area under the curve (AUC) and a positive likelihood ratio and that combined CpG 76–79 showed the highest AUC among all combinations. AUCs of consecutive CpG combinations tended to be higher for CpG 74–82 in exon 1 than for CpG 83–89 in intron 1. In conclusion, the methylation status at CpG sites in exon 1 was strongly associated with TVR reduction in glioblastoma.

A transparent, optically observable microfluidic device for microwave-induced chemical reactions using 24.125 GHz ISM band incorporating was developed. The microfluidic channels can pass through gaps in the post-wall waveguide. The post-wall waveguide allows microwave irradiation to be applied to designed area of the microfluidic channel. In this study, Polydimethylsiloxane (PDMS), commonly used in microfluidic devices, was used as the microwave waveguide material. A glass plate sputtered with indium tin oxide was used to shield microwave leakage to the top and bottom. 4 W of microwave input power was used to heat ethylene glycol, which is used as a solvent in chemical synthesis, in the channels of the fabricated device, and a temperature rise to 100 °C was observed in 70 s. We believe that the use of PDMS as a waveguide material will facilitate observation during microwave irradiation using optics and combination with other microreactors.

Superoxide dismutase 1 (Sod1) plays pivotal roles in antioxidation via accelerating the conversion of superoxide anion radicals into hydrogen peroxide, thus inhibiting the subsequent radical chain reactions. While Sod1 deficient cells inevitably undergo death in culture conditions, Sod1-knockout (KO) mice show relatively mild phenotypes and live approximately two years. We hypothesized that the presence of abundant levels of ascorbic acid (AsA), which is naturally produced in mice, contributes to the elimination of reactive oxygen species (ROS) in Sod1-KO mice. To verify this hypothesis, we employed mice with a genetic ablation of aldehyde reductase (Akr1a), an enzyme that is involved in the biosynthesis of AsA, and established double knockout (DKO) mice that lack both Sod1 and Akr1a. Supplementation of AsA (1.5 mg/ml in drinking water) was required for the DKO mice to breed, and, upon terminating the AsA supplementation, they died within approximately two weeks regardless of age or gender. We explored the etiology of the death from pathophysiological standpoints in principal organs of the mice. Marked changes were observed in the lungs in the form of macroscopic damage after the AsA withdrawal. Histological and immunological analyses of the lungs indicated oxidative damage of tissue and activated immune responses. Thus, preferential oxidative injury that occurred in pulmonary tissues appeared to be primary cause of the death in the mice. These collective results suggest that the pivotal function of AsA in coping with ROS in vivo, is largely in pulmonary tissues that are exposed to a hyperoxygenic microenvironment.

Next-generation sequencing-based comprehensive genomic profiling test (CGPT) enables clinicians and patients to access promising molecularly targeted therapeutic agents. Approximately 10% of patients who undergo CGPT receive an appropriate agent. However, its coverage of glioma patients is seldom reported. The aim of this study was to reveal the comprehensive results of CGPT in glioma patients in our institution, especially the clinical actionability. We analyzed the genomic aberrations, tumor mutation burden scores, and microsatellite instability status. The Molecular Tumor Board (MTB) individually recommended a therapeutic agent and suggested further confirmation of germline mutations after considering the results. The results of 65/104 patients with glioma who underwent CGPTs were reviewed by MTB. Among them, 12 (18.5%) could access at least one therapeutic agent, and 5 (7.7%) were suspected of germline mutations. A total of 49 patients with IDH-wildtype glioblastoma showed frequent genomic aberrations in the following genes: TERT promoter (67%), CDKN2A (57%), CDKN2B (51%), MTAP (41%), TP53 (35%), EGFR (31%), PTEN (31%), NF1 (18%), BRAF (12%), PDGFRA (12%), CDK4 (10%), and PIK3CA (10%). Since glioma patients currently have very limited standard treatment options and a high recurrence rate, CGPT might be a facilitative tool for glioma patients in terms of clinical actionability and diagnostic value.

Members of Clibanarius virescens show considerable intraspecific colour variation, including colouration of the second/third pereopods (green/white) and the dactyls of the second/third pereopods (with or without dark bands/patches). However, factors inducing these colour variations have not yet been elucidated. Here, we investigated the occurrence of colour variation in this species with particular emphasis on change of colouration associated with growth stage and region in specimens from tropical/subtropical to warm temperate areas in the Indo–West Pacific, including evidence from molecular phylogeny based on the cytochrome c oxidase subunit I (COI). We have, then, clarified that the colouration on the pereopod dactyls gradually changed from solid colour (yellow/white) to having dark-coloured area(s) or transverse band(s) as a result of the growth stage. The frequency of occurrence of the solid colour dactyls was higher than those of other colour types in tropical regions. Our results also indicated that the white ambulatory leg type was the colouration type that was frequently seen in juvenile stages. However, significant genetic differences were not detected between each colouration determined by molecular analysis of samples from 14 localities in the Indo-West Pacific region; in contrast, two genetically differentiated regional populations (North Australia; Phuket, Thailand; and Lombok, Indonesia) were detected. The present study, therefore, emphasizes the necessity for further study on the colour variation of marine animals focusing on growth stages and regional differences, with molecular data to facilitate the research on adaptation and/or speciation, especially in geographically widely distributed species.

Background Secondary pneumothorax after chemotherapy for a malignant pulmonary tumor is a complication from a large cavity causing refractory pneumothorax. Case presentation A 61-year-old man was referred due to prolonged air leakage from a pulmonary cavity that developed after treatment for pulmonary metastases from renal cell carcinoma. As air leakage continued after thoracic drainage and endobronchial occlusion, we planned thoracoscopy-assisted surgery. Intraoperatively, a large cavity opening to the pulmonary cavity was found in the left upper lobe. As it was difficult to repair the fistula using staplers or direct sutures because the pleura around the cavity was thick and hard, we attempted to plombage the cavity with a pericardial fat pad. After the operation, air leakage immediately disappeared and no recurrence of the pneumothorax was found. Conclusion This novel method can be useful to seal a large bronchopleural fistula that causes refractory pneumothorax.

The patellogastropod limpet genus Nipponacmea is widely distributed in Japan and adjacent East Asia. Species identification within Nipponacmea is challenging due to the high variation in shell morphology. In this study, we examined the taxonomy of this genus represented by nine nominal species from 43 localities (including type localities). Results of the molecular phylogenetic analysis revealed that: (1) N. gloriosa, the sole species in this genus inhabiting the subtidal zone, represents the most basal independent branch; (2) the remaining species are divided into two large clades with lower- and higher-apex shell profiles; and (3) the high-apex morphology was derived from the low-apex type. The terminal clades defined using the molecular data were consistent with nine morphospecies and had 100% bootstrap values, strongly supporting the conventional taxonomy of Nipponacmea. Although morphological similarities do not always reflect phylogeny, the set of morphological characters used in the current taxonomy were proven to be adequate for diagnosis. In conclusion, this study provided solid evidence to uphold the monophyly of known species of Nipponacmea in Japan and demonstrated the usefulness of morphological characters for species diagnosis.

Diacylglycerol (DG) is unique in lipid metabolism because it serves not only as an intermediate product for triglyceride synthesis, but also as a signaling molecule that activates proteins containing DG-responsive elements, such as protein kinase C. Consequently, DG acts as a hub between energy metabolism and intracellular signaling. Of DG metabolizing pathways, DG kinase (DGK) phosphorylates DG to produce phosphatidic acid, which also serves as a second messenger. Several lines of evidence suggest that DGK is deeply involved in metabolic diseases such as obesity and insulin resistance. Of DGK isozymes, DGKε is simplest in terms of structure, but it is characterized by substrate specificity toward arachidonoyl-DG. Recently, we have reported that DGKε deficiency promotes adipose tissue remodeling in mice during the course of high fat diet (HFD) feeding regimen including obesity, insulin resistance, and beige adipogenesis. DGKε ablation engenders altered expression of other lipid metabolizing enzymes, including adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL), and diacylglycerol acyltransferase (DGAT). Subcellular localization of DGKε in the endoplasmic reticulum suggests involvement of this isozyme in lipid energy homeostasis. This review presents current findings of DGKε in lipid-orchestrated pathophysiology, especially unique phenotypes of DGKε-knockout mice in the early and late stages of obesogenic conditions.

Diacylglycerol kinase (DGK) is a lipid kinase that converts diacylglycerol (DG) to phosphatidic acid (PA). Since both DG and PA serve as intracellular second messenger molecules, DGK plays a pivotal role in balancing these two signaling pathways. Of the DGK family, DGKη is classified as a type II DGK. Reportedly, DGKη is expressed ubiquitously through mammalian tissues and cells. Previous studies using cDNA transfection methods reported cytoplasmic localization of DGKη in cultured human cells. However, subcellular localization of native protein is still unknown. Recently, we established a human DGKη-specific monoclonal antibody, DhMab-4. In this study, we examined subcellular localization of native protein of DGKη using DhMab-4 by immunocytochemistry in human cultured cells.

Background Brain metastases (BM) to the cerebellum account for approximately 20% of all of BM. From the results of JCOG0504 clinical trial, observation with salvage SRS (stereotactic radiosurgery) has been established as a standard therapy for a patient after complete resection of BM. However, we often experience early local recurrence after resection of cerebellar BM, therefore, we retrospectively analyzed them to consider the appropriate timing of SRS. Methods We extracted surgical cases with cerebellar BM from January 2015 to March 2022 and analyzed the patient characteristics, treatment content, postoperative intracranial progression-free survival (IC-PFS) and postoperative overall survival (OS). Results A total of 57 cases were enrolled (male: female = 43.9%: 56.1%) in this study. The median age was 61.9 years (51.9-68.2). The common primary lesions were NSCLC (31.6%), colorectal cancer (19.3%), and breast cancer (17.5%). The IC-PFS was 4.5 months (2.6-5.7) and the OS was 11.6 months (9.1-21.8), but the local recurrent rate within 3 months was 43.4%. The rate of patients with leptomeningeal metastases (LMM) at the first recurrence was 26.3%, significantly higher than 7.7% in JCOG0504 (p=0.000224). In the postoperative observation group (no SRS within a month), the OS was 25.3 months (11.6-NA), which was longer than the 15.6 months (11.4-20.8) of JCOG0504. RT was performed in 18 patients at the recurrence (SRS: 15, WBRT (whole-brain radiotherapy): 9). SRS was the only factor that prolonged the OS (25.2 vs. 6.1 months (p=0.0212)). Furthermore, in the patients who underwent radiotherapy within 1 month after surgery, postoperative SRS (10.2 vs. 3.1 months, p=0.00437), and postoperative SRS to the resection cavity (11.0 vs. 3.7mo, p=0.0012) significantly prolonged the OS. Conclusion Since cerebellar BM has high recurrence and frequent LMM, early SRS for a resected cavity after surgery might lead to more favorable outcomes in cerebellar BM patients.

The patellogastropod limpet genus

Background: Several studies have reported that gross total resection contributes to improved prognosis in patients with butterfly glioblastoma (bGBM). However, it sometimes damages the corpus callosum and cingulate gyrus, leading to severe neurological complications. Case Description: We report two cases of bGBM that was safely and maximally resected using brief and exact awake mapping after general anesthesia. Two patients had butterfly tumors in both the frontal lobes and the genu of the corpus callosum. Tumor resection was first performed on the nondominant side under general anesthesia to shorten the resection time and maintain patient concentration during awake surgery. After that, awake surgery was performed for the lesions in the dominant frontal lobe and genu of the corpus callosum. Tumor resection was performed through minimal cortical incisions in both frontal lobes. Postoperative magnetic resonance imaging showed gross total resection, and the patients had no chronic neurological sequelae, such as akinetic mutism and abulia. Conclusion: bGBM could be safely and maximally resected by a combination of asleep and brief awake resection, which enabled patients to maintain their attention to the task without fatigue, somnolence, or decreased attention. The bilateral approach from a small corticotomy can avoid extensive damage to the cingulate gyrus.

Continuing molecular and morphological investigations of a limpet recently reported as introduced to Sicily in the Mediterranean Sea have revealed it to be an undescribed species of Lottia Gray, 1833, a genus that is native to Sri Lanka and vicinity in the northern Indian Ocean. The genetic and morphological features of the limpets compared from Sicily and Sri Lanka correspond so closely that we do not hesitate to describe it collectively as Lottia iani n. sp., with type locality of Tangalle on the coast of Sri Lanka. Much is still unknown about its native range, when and how it arrived in Sicily, whether it is capable of impacting native species, and whether its introduction might be more widespread in the Mediterranean than just the particular eastern Sicilian localities where it has been observed. It is plausible that the introduction of L. iani to Sicily could have resulted from recent expansions to the Suez Canal, which is known to have led to other species introductions in recent years. Other possibilities include an introduction related to shipping or mariculture activities. However it managed to arrive, this limpet species has clearly been able to establish a self-recruiting local population at particular Sicilian localities with rocky volcanic shores.

Patellogastropoda is an ecologically widely diversified group living on various substrata. Juveniles of Lottia tenuisculpta live on rocks and also on the shell of trochid snails, from which host th...

Diacylglycerol kinase (DGK) is an enzyme that converts diacylglycerol (DG) to phosphatidic acid (PA). As both DG and PA serve as lipidic second messengers, DGK plays a pivotal role in controlling the balance of two signaling pathways mediated by DG and PA in cellular functions. DGKζ, one member of the mammalian DGK family, is reported to contain a nuclear localization signal, which suggests its functional role in the nucleus. Previously, morphological studies using tagged expression vectors and immunostaining of rat tissues or cells have revealed that DGKζ localizes mainly to the nucleus. However, a limited number of studies reported the detailed localization of native protein of DGKζ in human tissues and cells. In this study, we developed a novel anti-human DGKζ monoclonal antibody, DzMab-1, which is very advantageous in immunocytochemistry of human cultured cells.

Continuous long-term monitoring is important for detecting ecological changes and understanding their causes, including anthropogenic impacts such as climate changes and eutrophication. Nonetheless, such long-term population studies have been rare, especially for sea urchins, which can affect community dynamics owing to their extensive herbivory and large population fluctuations. Here we present a long-term (from 1963 to 2014) dynamics of sea urchins in a fixed quadrat on a lower intertidal rocky flat in Hatakejima Island, southern Japan. We also conducted a complementary survey over the entire island approximately every five years from 1975 to 2013, and a 41-year assay for developmental abnormality of the sea urchin Heliocidaris crassispina using water adjacent to the island. The abundance of three commonest species in this area (H. crassispina, Echinostrephus molaris, and Echinometra spp.) and the richness of urchin species showed large variations, with high numbers in the 1960s and 1970s, followed by an abrupt decline in the late 1970s or early 1980s, and a gradual recovery subsequently. The species abundance and richness in the entire island survey showed good correlations with those in the quadrat census. Statistical analyses indicated that increasing water temperature and red tides were the major factors influencing the dynamics of abundance and species richness. Our studies reveal that anthropogenic environmental changes influence the long-term dynamics in abundance and richness of sea urchins.

True limpets of the genus Nacella (Patellogastropoda: Nacellidae) include at least 11 nominal species distributed in different provinces of the Southern Ocean. Here, we present new molecular analyses and a comprehensive morphological revision of Nacella confirming the validity of all the currently recognized species, but with important amendments to the published distributions of several. We also show that specimens collected along two fjords in the Beagle Channel, southern South America, constitute a new taxon: Nacella yaghana sp. nov. The phylogenetic position of this new taxon in our molecular tree reveals that it represents an ancestral South American lineage of Nacella, markedly separated from the previously recognized and evolutionary recent Magellanic radiation. In addition, this new Nacella species was clearly distinguished from the rest of the species by morphological comparisons, including shell characteristics, radular-tooth shape and configuration, as well as by the coloration pattern of the mantle tentacles. This study provides new evidence about the evolutionary history of this important Southern Ocean, marine, near-shore benthic group and the role of the Antarctic Circumpolar Current in the biogeography of the genus.

To investigate the effect of alteplase, either combined with stent-retriever thrombectomy or a direct aspiration first pass technique (ADAPT), in patients with large-vessel occlusion stroke. This was a retrospective post hoc analysis of data from The Direct Mechanical Thrombectomy in Acute LVO Stroke (SKIP) study. Patients were divided into two groups according to the first-line thrombectomy technique: stent-retriever and ADAPT. Each group was further divided into two subgroups, namely MT and MT + alteplase. The procedural outcomes, such as first pass effect (FPE) ratio and number of passes, were evaluated. The clinical outcomes included mRS score at 3 months. A total of 180 patients were included (116 in the stent-retriever group and 64 in the ADAPT group). No interaction was detected between the first-line technique and alteplase administration. In the stent-retriever group, after adjusting for factors associated with FPE, the adjusted odds ratio (95% confidence interval) of FPE of the MT + alteplase subgroup versus the MT subgroup was 3.57 (1.5–8.48) and in the ADAPT group it was 1.35 (0.37–4.91). With alteplase, the number of passes decreased with adjusted odds ratios of 0.59 (0.37–0.93) in the stent-retriever group but not in the ADAPT group. In both first-line technique groups, clinical outcomes did not differ between subgroups. In the SKIP study, alteplase administration was associated with increased FPE when combined with stent-retriever thrombectomy, but not with ADAPT. We found no differences in the clinical outcomes.

Diacylglycerol kinase (DGK) constitutes a family of enzymes that phosphorylate diacylglycerol to phosphatidic acid (PA). These lipids serve as second messengers, thereby activating distinct downstream cascades and different cellular responses. Therefore, DG-to-PA conversion activity induces a phase transition of signaling pathways. One member of the family, DGKζ, is involved closely with stress responses. Morphological data showing that DGKζ localizes predominantly to the nucleus and that it shuttles between the nucleus and the cytoplasm implicate DGKζ in the regulation of transcription factors during stress responses. Tumor suppressor p53 and NF-κB are major stress-responsive transcription factors. They exert opposing effects on cellular pathophysiology. Herein, we summarize DGKζ catalytic activity-dependent and -independent regulatory mechanisms of p53 and NF-κB transactivation activities, including p53 degradation and NF-κB nuclear translocation. We also discuss how each component of DGKζ-interacting protein complex modulates the specificity and selectivity of target gene expression.

Objectives Selection criteria for palliative limited surgery in patients with non-small cell lung cancer (NSCLC) can vary by institution or surgeon. We retrospectively reviewed outcomes of poor-risk patients who underwent palliative segmentectomy (PS), using the National Clinical Database Risk Calculator (RC). Methods We retrospectively analyzed medical records of patients with NSCLC tumors ≥ 20 mm and consolidation/tumor ratios ≥ 0.5 on computed tomography, who underwent PS from January 2009 to March 2016. Median follow-up time was 47 months (range 2–102 months). Results We enrolled 67 patients (median age: 73.0 years), of whom 54 received thoracoscopic surgery and 28 received medial lymph-node dissection. The RC’s mean predictive probability rate for perioperative mortality or severe complications was 7.1%. Of the 67 patients, 24 patients (43.0%) suffered post-surgical complications, including 2 (3%) who died in hospital; 17 eventually suffered NSCLC recurrences and/or metastases, 11 eventually died from NSCLC, and 17 died from other diseases. Five-year overall survival (OS) was 59.4%. When the patients were divided into high-risk (HR) and low-risk (LR) groups based on the RC, 5-year OS was significantly less in the HR group (43.9%) than in the LR group (82.2%; P Conclusion The RC, which was developed primarily to determine perioperative risk, can predict long-term prognosis for compromised patients who undergo PS.

We report the diversity, distribution and taxonomy of patellogastropod species from the entire mainland Indian coastline. A total of 77 coastal areas with rocky outcrops in the sandy shoreline, covering the entire mainland Indian coastline, were surveyed seasonally (four times a year) during December 2014 to December 2016. Coastal habitat characteristics and the diversity of all groups of limpets from the intertidal areas were recorded. Specimens were identified by external morphological shell and gill characteristics, by the colour of the foot, mantle fringe and radular characteristics. Ecological aspects like habitat preference were also used to distinguish species. Existing diagnostic characters were amended with newer characteristics emerged in our study. Seven species from two families, Nacellidae and Lottiidae, were found along the mainland Indian coastline. Each species is present with several polymorphic forms with different colour banding patterns. Our study revealed distinct variations in the radular teeth at the genus level, but less differences within genera. Patellogastropod diversity was greater along the west coast of India and only Cellana rota was available throughout the mainland Indian coastline.

The tiny epizoic limpet Lottia tenuisculpta lives on rocky surfaces and shells of the snails Omphalius nigerrimus and Reishia clavigera. The movement patterns of the limpet on host snails was obser...

Diacylglycerol kinase (DGK) phosphorylates diacylglycerol (DG) to produce phosphatidic acid (PA). Since both DG and PA serve as lipidic second messengers, DGK plays a pivotal role in regulating the balance of two signaling pathways mediated by DG and PA in cellular functions. Reportedly, DGKγ, one of the 10 mammalian DGK isozymes, is involved in leukemic cell differentiation, mast cell function, and membrane traffic. Transfection studies using tagged expression vectors and immunohistochemistry on rat tissues revealed that DGKγ localizes to the cytoplasm, plasma membrane, and Golgi apparatus. However, a limited number of studies reported the detailed localization of native protein of DGKγ in human tissues and cells. In this study, we developed a novel anti-DGKγ monoclonal antibody, DgMab-6, which is very useful in immunocytochemistry of human cultured cells.

Recently developed molecular targeted therapies such as bevacizumab (BEV; Avastin) therapy have therapeutic efficacy for glioblastoma. However, it is difficult to distinguish between a tumor response and nonenhancing tumor progression with conventional magnetic resonance imaging (MRI) after BEV administration. Here we present a recurrent glioblastoma case in which 11C-methionine positron emission tomography (MET-PET) provided useful information for detecting tumor recurrence after complete remission, as assessed by the Response Assessment in Neuro-Oncology criteria. A 47-year-old male with a left frontal lobe glioblastoma experienced recurrence 6 months postoperatively. We administered BEV concomitantly with temozolomide, subsequent to gamma knife surgery. Two months after starting BEV, complete remission was obtained. MET uptake on PET gradually decreased and had nearly disappeared 4 months after initiating BEV. No enhanced area was seen on MRI for 17 months after BEV initiation. Nevertheless, MET-PET revealed recurrence, visualized as nonenhancing tumor progression. MET-PET provides useful information for detecting glioblastoma recurrence, which lacks contrast enhancement on MRI after BEV therapy.

During our survey on the intertidal hermit crab fauna on Amami Oshima Island and Okinawa Island, southwestern Japan, we collected specimens identified asClibanariusspecies by their morphological features, but that had an unknown coloration for the genus: they have whitish ambulatory legs and a whitish shield. For species identification of these specimens, we observed their coloration and morphology and also performed phylogenetic analyses using the genes mitochondrial cytochrome c oxidase I (COI) and nuclear histone H3. The analysis has shown that these specimens are to be identified asClibanarius virescens(Krauss, 1843).

BACKGROUND Central nervous system germ cell tumor (CNSGCT) is a rare pediatric brain tumor. However, they are found at a relatively high incidence in East Asia. Germinoma is sensitive toward radiotherapy and chemotherapy; however, non-germinoma GCTs (NGGCT) often show poor response. Some cases are a mixture of germinoma and NGGCT (mixed GCT), and they sometimes change histological subtypes at recurrence. Previous report demonstrated that a germinoma and NGGCT component within the same mixed GCT tissue shared the same gene mutation, whereas the genome-wide methylation profiles were distinct from each other. The methylation profiles of germinoma was similar to the primordial germ cells (PGC) at the migration phase, supporting a model that PGC is the cell of origin for CNSGCT. However, tumor heterogeneity hinder information of the mixed bulk RNA-sequence data, causing difficulty in elucidating the mechanism of tumor development. The purpose of this study was to investigate the tumor cells subpopulations at the resolution of individual cells by single-cell RNA-seq. RESULTS Fresh surgical tumor tissue was immediately dissociated mechanically and enzymatically. Tumor cells are separated from CD45-labelled lymphocytes by FACS, and libraries were generated by Chromium Single cell 3’ Reagent Kit. Total of 11 tumor samples were collected and sequenced. Unsupervised Clustering showed individual clusters. One of the clusters had high expression of Oct-4, which is a marker of germinoma. The other clusters showed different subtypes of cells representing the heterogeneity of CNSGCT. Further analysis including a pseudo-time course analysis is underway to identify the lineage of tumor cell development.

Carboxylesterase (CES) plays an important role in the hydrolysis metabolism of ester-type drugs and prodrugs. In this study, we investigated the change in the hydrolysis rate of hCE1 by focusing on the steric hindrance of the ester structure and the electron density. For 26 kinds of synthesized indomethacin prodrugs, the hydrolytic rate was measured in the presence of human liver microsomes (HLM), human small intestine microsomes (HIM), hCE1 and hCE2. The synthesized prodrugs were classified into three types: an alkyl ester type that is specifically metabolized by hCE1, a phenyl ester type that is more easily metabolized by hCE1 than by hCE2, and a carbonate ester type that is easily metabolized by both hCE1 and hCE2. The hydrolytic rate of 1-methylpentyl (hexan-2-yl) ester was 10-times lower than that of 4-methylpentyl ester in hCE1 solution. hCE2 was susceptible to electron density of the substrate, and there was a difference in the hydrolysis rate of up to 3.5-times between p-bromophenyl ester and p-acetylphenyl ester. By changing the steric hindrance and electron density of the alkoxy group, the factors that change the hydrolysis rate by CES were elucidated.

Very little is known about the evolution of molluskan shell pigments, although Mollusca is a highly diverse, species rich, and ecologically important group of animals comprised of many brightly colored taxa. The marine snail genus Clanculus was chosen as an exceptional model for studying the evolution of shell color, first, because in Clanculus margaritarius and Clanculus pharaonius both shell and foot share similar colors and patterns; and second, because recent studies have identified the pigments, trochopuniceus (pink‐red), and trochoxouthos (yellow‐brown), both comprised of uroporphyrin I and uroporphyrin III, in both shell and colored foot tissue of these species. These unusual characteristics provide a rare opportunity to identify the genes involved in color production because, as the same pigments occur in the shell and colored foot tissue, the same color‐related genes may be simultaneously expressed in both mantle (which produces the shell) and foot tissue. In this study, the transcriptomes of these two Clanculus species along with a third species, Calliostoma zizyphinum, were sequenced to identify genes associated with the synthesis of porphyrins. Calliostoma zizyphinum was selected as a negative control as trochopuniceus and trochoxouthos were not found to occur in this species. As expected, genes necessary for the production of uroporphyrin I and III were found in all three species, but gene expression levels were consistent with synthesis of uroporphyrins in mantle and colored foot tissue only in Clanculus. These results are relevant not only to understanding the evolution of shell pigmentation in Clanculus but also to understanding the evolution of color in other species with uroporphyrin pigmentation, including (mainly marine) mollusks soft tissues and shells, annelid and platyhelminth worms, and some bird feathers.