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Ribosome-associated quality control (RQC) pathway is responsible for degradation of nascent polypeptides in aberrantly stalled ribosomes, and its defects may lead to neurological diseases. However, the underlying molecular mechanism of how RQC dysfunction elicits neurological disorders remains poorly understood. Here we revealed that neurons with knockout (KO) of ubiquitin ligase LTN1, a key gene in the RQC pathway, show developmental defects in neurons via upregulation of TTC3 and UFMylation signaling proteins. The abnormally enhanced TTC3 protein in Ltn1 KO neurons reduced further accumulation of translationally arrested products by preventing translation initiation of selective genes. However, the overaccumulated TTC3 protein in turn caused dendritic abnormalities and reduced surface-localized GABA A receptors during neuronal development. Ltn1 KO mice showed behavioral deficits associated with cognitive disorders, a subset of which were restored by TTC3 knockdown in medial prefrontal cortex. Together, the overactivated cellular compensatory mechanism against defective RQC through TTC3 overaccumulation induced synaptic and cognitive deficits. More broadly, these findings represent a novel cellular mechanism underlying neuronal dysfunctions triggered by exaggerated cellular stress response to accumulated abnormal translation products in neurons.

This study aims to create a terrain classification of Japan that allows both geomorphological and geoengineering classifications coexist without large contradictions and to distinguish landform elements even in urban plains which include noise associated with digital elevation models (DEMs). Because Japan is susceptible to natural disasters, we designed the classification to reflect the ground vulnerability of both alluvial plains and mountains through the application of terrain classification data to landslide susceptibility and seismic zoning. We updated an existing DEM-based terrain classification method for application in the high-resolution 30 m DEM. We used topographic measurements that do not amplify manmade unevenness or noise, which are usually the main problems associated with the use of high-resolution DEMs with high vertical accuracies. We selected the height above the nearest drainage (HAND), slope gradient, surface texture, and local convexity as geometric signatures, which were devised so as not to detect noise. Segment polygon data of terrain units were derived from the raster data of slope and HAND. The polygon data were classified into 40 clusters using the attributes of slope, HAND, and surface texture; then they were grouped into 16 legends following comparisons with the existing geological and geomorphological maps and supplementary reclassification by HAND and local convexity. The derived terrain classification, except for manmade cuts and fills, showed similarities with the existing expert-driven maps and some association with areas where shallow landslides or floods frequently occur. Based on a trial in California using a 30 m DEM, we concluded that the proposed method can be adopted in other regions outside of Japan.

Familial juvenile hyperuricemic nephropathy (FJHN) is an autosomal dominant disorder caused by mutations in UMOD. Here we studied effects of genetic expression and pharmacological induction of Hsp70 on the UMOD mutants C112Y and C217G.We expressed wild type (WT), C112Y and C217G in HEK293 cells and studied their maturation and cellular damage using western blot and flow cytometry.Expression of C112Y or C217G increased pro-apoptotic proteins, decreased anti-apoptotic proteins, and induced cellular apoptosis as examined by annexin V staining and flow cytometry. Overexpression of Hsp70 or administration of an Hsp70 inducer geranylgeranylacetone (GGA) promoted maturation of the mutant proteins, increased their secreted forms, normalized the levels of pro- and anti-apoptotic proteins and suppressed apoptosis.These findings indicated that Hsp70 enhanced maturation of C112Y and C217G and reduced cellular apoptosis, suggesting that Hsp70 induction might be of a therapeutic value for treatment of FJHN.

Recent genetic approaches have demonstrated that genetic factors contribute to the pathologic origins of neuropsychiatric disorders. Nevertheless, the exact pathophysiological mechanism for most cases remains unclear. Recent studies have demonstrated alterations in pathways of protein homeostasis (proteostasis) and identified several proteins that are misfolded and/or aggregated in the brains of patients with neuropsychiatric disorders, thus providing early evidence that disrupted proteostasis may be a contributing factor to their pathophysiology. Unlike neurodegenerative disorders in which massive neuronal and synaptic losses are observed, proteostasis impairments in neuropsychiatric disorders do not lead to robust neuronal death, but rather likely act via loss- and gain-of-function effects to disrupt neuronal and synaptic functions. Furthermore, abnormal activation of or overwhelmed endoplasmic reticulum and mitochondrial quality control pathways may exacerbate the pathophysiological changes initiated by impaired proteostasis, as these organelles are critical for proper neuronal functions and involved in the maintenance of proteostasis. This perspective article reviews recent findings implicating proteostasis impairments in the pathophysiology of neuropsychiatric disorders and explores how neuronal and synaptic functions may be impacted by disruptions in protein homeostasis. A greater understanding of the contributions by proteostasis impairment in neuropsychiatric disorders will help guide future studies to identify additional candidate proteins and new targets for therapeutic development.

An amendment to this paper has been published and can be accessed via the original article.

Neurodegenerative disorders such as Huntington’ disease and frontotemporal lobar degeneration (FTLD) are often caused by aggregation of causative proteins and co-morbid with mental disorders of unknown origin. Although toxic protein aggregates may entrap proteins involved in psychiatric disorders and elicit deleterious effects, candidate mechanisms for this co-aggregation hypothesis remain unknown. Here we show a novel cytosolic protein aggregate in FTLD brain composed of DISC1, a biological mediator for mental illnesses and TDP-43, the major protein constituent of insoluble inclusions in FTLD. FTLD model mice with DISC1/TDP-43 co-aggregates in frontal cortex showed hyperactivity and social interaction deficits that were rescued by exogenous DISC1 expression. At the cellular level, the amounts of newly synthesized proteins in dendrites are reduced in the neurons that lack DISC1 expression or contain DISC1/TDP-43 co-aggregates. These results support our novel hypothesis that co-aggregation of risk factors for psychiatric diseases may compromise local translation in dendrites and mental conditions.

Myocardial ischemia/reperfusion injury worsens in the absence of nitric oxide synthase (NOS). Cilnidipine, a Ca2+ channel blocker, has been reported to activate endothelial NOS (eNOS) and increases nitric oxide (NO) in vascular endothelial cells. We examined whether pretreatment with cilnidipine could attenuate cardiac cell deaths including apoptosis caused by hypoxia/reoxygenation (H/R) injury. HL-1 mouse atrial myocytes as well as H9c2 rat ventricular cells were exposed to H/R, and cell viability was evaluated by an autoanalyzer and flow cytometry; eNOS expression, NO production, and electrophysiological properties were also evaluated by western blotting, colorimetry, and patch clamping, respectively, in the absence and presence of cilnidipine. Cilnidipine enhanced phosphorylation of eNOS and NO production in a concentration-dependent manner, which was abolished by siRNAs against eNOS or an Hsp90 inhibitor, geldanamycin. Pretreatment with cilnidipine attenuated cell deaths including apoptosis during H/R; this effect was reproduced by an NO donor and a xanthine oxidase inhibitor. The NOS inhibitor L-NAME abolished the protective action of cilnidipine. Pretreatment with cilnidipine also attenuated H9c2 cell death during H/R. Additional cilnidipine treatment during H/R did not significantly enhance its protective action. There was no significant difference in the protective effect of cilnidipine under normal and high Ca2+ conditions. Action potential duration (APD) of HL-1 cells was shortened by cilnidipine, with this shortening augmented after H/R. L-NAME attenuated the APD shortening caused by cilnidipine. These findings indicate that cilnidipine enhances NO production, shortens APD in part by L-type Ca2+ channel block, and thereby prevents HL-1 cell deaths during H/R.

Local governments often rely on hazard maps to plan for and respond to local natural disasters. These maps often rely on the use of many colors, but their exact color scheme and style differs in each area. As a result, we hypothesize that some users, specifically colorblind individuals, may have difficulty correctly understanding the information on some of these hazard maps. In this study, we test that hypothesis by conducting a survey of Japanese liquefaction hazard maps and their visual accessibility. To this end, we first undergo a survey of the color schemes used in these maps and investigate whether they are easily understood by people with colorblindness. We next specifically analyze several maps we deem particularly problematic in terms of color scheme and visibility, using these as case studies to discuss issues with accessibility and to summarize possible countermeasures. Our survey found that liquefaction hazard maps use one of three main color schemes: “diverging color scheme,” “Sequential cold colors,” or “sequential warm colors.” However, while there were issues with several maps, including difficulty reading the background map or correctly understanding the risk of liquefaction, these difficulties were not related to the color scheme used. To improve the accessibility of hazard maps, therefore, is necessary to create a unified manual that contains the following information: an examination of colors, the utilization of a universal design check tool, and the use of GIS vector data.

Dysfunctional mTOR signaling is associated with the pathogenesis of neurodevelopmental and neuropsychiatric disorders. However, it is unclear what molecular mechanisms and pathogenic mediators are involved and whether mTOR-regulated autophagy continues to be crucial beyond neurodevelopment. Here, we selectively deleted Atg7 in forebrain GABAergic interneurons in adolescent mice and unexpectedly found that these mice showed a set of behavioral deficits similar to Atg7 deletion in forebrain excitatory neurons. By unbiased quantitative proteomic analysis, we identified γ-aminobutyric acid receptor-associated protein-like 2 (GABARAPL2) to differentially form high-molecular weight species in autophagy-deficient brains. Further functional analyses revealed a novel pathogenic mechanism involving the p62-dependent sequestration of GABARAP family proteins, leading to the reduction of surface GABAA receptor levels. Our work demonstrates a novel physiological role for autophagy in regulating GABA signaling beyond postnatal neurodevelopment, providing a potential mechanism for the reduced inhibitory inputs observed in neurodevelopmental and neuropsychiatric disorders with mTOR hyperactivation.

De novo protein synthesis by the ribosome and its multitude of co-factors must occur in a tightly regulated manner to ensure that the correct proteins are produced accurately at the right time and, in some cases, also in the proper location. With novel techniques such as ribosome profiling and cryogenic electron microscopy, our understanding of this basic biological process is better than ever and continues to grow. Concurrently, increasing attention is focused on how translational regulation in the brain may be disrupted during the progression of various neurological disorders. In fact, translational dysregulation is now recognized as the de facto pathogenic cause for some disorders. Novel mechanisms including ribosome stalling, ribosome-associated quality control, and liquid-liquid phase separation are closely linked to translational regulation, and may thus be involved in the pathogenic process. The relationships between translational dysregulation and neurological disorders, as well as the ways through which we may be able to reverse those detrimental effects, will be examined in this review.

Autophagy regulates social behaviors by affecting the ability of brain cells to receive inhibitory signaling., Dysfunctional mTOR signaling is associated with the pathogenesis of neurodevelopmental and neuropsychiatric disorders. However, it is unclear what molecular mechanisms and pathogenic mediators are involved and whether mTOR-regulated autophagy continues to be crucial beyond neurodevelopment. Here, we selectively deleted Atg7 in forebrain GABAergic interneurons in adolescent mice and unexpectedly found that these mice showed a set of behavioral deficits similar to Atg7 deletion in forebrain excitatory neurons. By unbiased quantitative proteomic analysis, we identified γ-aminobutyric acid receptor–associated protein-like 2 (GABARAPL2) to differentially form high–molecular weight species in autophagy-deficient brains. Further functional analyses revealed a novel pathogenic mechanism involving the p62-dependent sequestration of GABARAP family proteins, leading to the reduction of surface GABAA receptor levels. Our work demonstrates a novel physiological role for autophagy in regulating GABA signaling beyond postnatal neurodevelopment, providing a potential mechanism for the reduced inhibitory inputs observed in neurodevelopmental and neuropsychiatric disorders with mTOR hyperactivation.

In this study, we investigated the fabrication of porous chitins with continuous channel substructure by regeneration from gels with CaBr2·2H2O/methanol solution. After rapidly removing methanol from the gels, the products were immersed in methanol, followed by washing out CaBr2 with water and lyophilization to obtain regenerated chitins with inter-connected continuous pore morphology. Scanning electron microscopic results supported that the materials were consisted of continuous substructures of porous channels. The materials were further characterized by SEM-EDX and XRD measurements. The mechanical property and water absorbability were also evaluated. The plausible mechanism for the formation of the inter-connected pore morphology during the regeneration procedure was discussed.

In this study, we investigated the preparation of chitin nanofiber (CNF)-reinforced cellulose films through stepwise regeneration procedures from the respective ion gels with ionic liquids. Self-assembled CNF dispersions were prepared by regeneration from the chitin ion gel with the ionic liquid, 1-allyl-3-methylimidazolium bromide, using methanol, followed by dilution with adjusted amounts of methanol. Cellulose ion gels with the ionic liquid, 1-butyl-3-methylimidazolium chloride, were then prepared, soaked in the CNF dispersions, and centrifuged to simultaneously occur regeneration of cellulose and compatibilization with the CNFs. Soxhlet extraction with methanol and subsequent drying of the resulting materials gave the CNF/cellulose composite films. The IR and SEM results of the films indicated the presence of CNFs not only on the surfaces of the films but also inside the films. Powder X-ray diffraction patterns showed the amorphous structure of the cellulose in the film. Tensile testing of the films suggested the reinforcing effect of the CNFs on the mechanical properties of the films.

Metagenomes contain the DNA from many microorganisms, both culturable and non-culturable, and are a potential resource of novel genes. In this study, a 5.2-Gb metagenomic DNA library was constructed from a soil sample (artificially polluted with four aromatic compounds, i.e., biphenyl, phenanthrene, carbazole, and 3-chlorobenzoate) in Escherichia coli by using a broad-host-range cosmid vector. The resultant library was introduced into naphthalene-degrading Pseudomonas putida-derived strains having deficiencies in their naphthalene dioxygenase components, and indigo-forming clones on the indole-containing agar plates were screened. Cosmids isolated from 29 positive clones were classified by their various properties (original screening hosts, hosts showing indigo-forming activity, and digestion patterns with restriction enzymes), and six representative cosmids were chosen. Sequencing and in vitro transposon mutagenesis of the six cosmids resulted in the identification of genes encoding putative class B and D flavoprotein monooxygenases, a multicomponent hydroxylase, and a reductase that were responsible for the indigo-forming activity in the host cells. Among them, the genes encoding the multicomponent hydroxylase were demonstrated to be involved in phenol degradation. Furthermore, two genes encoding ring-cleavage dioxygenases were also found adjacent to the genes responsible for the indigo formation, and their functions were experimentally confirmed.

Background Neurodegenerative diseases involving protein aggregation often accompany psychiatric symptoms. Frontotemporal lobar degeneration (FTLD) associated with TAR DNA-binding protein 43 (TDP-43) aggregation is characterized by progressive neuronal atrophy in frontal and temporal lobes of cerebral cortex. Furthermore, patients with FTLD display mental dysfunction in multiple behavioral dimensions. Nevertheless, their molecular origin for psychiatric symptoms remains unclear. Methods In FTLD neurons and mouse models with TDP-43 aggregates, we examined coaggregation between TDP-43 and disrupted in schizophrenia 1 (DISC1), a key player in the pathology of mental conditions and its effects on local translation in dendrites and psychiatric behaviors. The protein coaggregation and the expression level of synaptic proteins were also investigated with postmortem brains from patients with FTLD (n = 6). Results We found cytosolic TDP-43/DISC1 coaggregates in brains of both FTLD mouse model and patients with FTLD. At the mechanistic levels, the TDP-43/DISC1 coaggregates disrupted the activity-dependent dendritic local translation through impairment of translation initiation and, in turn, reduced synaptic protein expression. Behavioral deficits detected in FTLD model mice were ameliorated by exogenous DISC1 expression. Conclusions Our findings reveal a novel role of the aggregate-prone TDP-43/DISC1 protein complex in regulating local translation, which affects aberrant behaviors relevant to multiple psychiatric dimensions.

Huntington's disease (HD) is a polyglutamine (polyQ) disease caused by aberrant expansion of the polyQ tract in Huntingtin (HTT). While motor impairment mediated by polyQ-expanded HTT has been intensively studied, molecular mechanisms for nonmotor symptoms in HD, such as psychiatric manifestations, remain elusive. Here we have demonstrated that HTT forms a ternary protein complex with the scaffolding protein DiSC1 and cAMP-degrading phosphodiesterase 4 (PDE4) to regulate PDE4 activity. We observed pathological cross-seeding between DiSC1 and mutant HTT aggregates in the brains of HD patients as well as in a murine model that recapitulates the polyQ pathology of HD (R6/2 mice). In R6/2 mice, consequent reductions in soluble DiSC1 led to dysregulation of DiSC1-PDE4 complexes, aberrantly increasing the activity of PDE4. Importantly, exogenous expression of a modified DiSC1, which binds to PDE4 but not mutant HTT, normalized PDE4 activity and ameliorated anhedonia in the R6/2 mice. We propose that cross-seeding of mutant HTT and DiSC1 and the resultant changes in PDE4 activity may underlie the pathology of a specific subset of mental manifestations of HD, which may provide an insight into molecular signaling in mental illness in general.

In this paper, we report acetylation of xanthan gum using acetic anhydride in an ionic liquid solvent, 1-butyl-3-methylimidazolium chloride (BMIMCl). Xanthan gum was dissolved with BMIMCl [2 % (w/w)] and the reaction was carried out in the presence of acetic anhydride (five equiv. for hydroxy groups in a repeating unit) with stirring the solution at elevated temperatures. The structures of xanthan gum acetates were confirmed by the 1H NMR and IR spectra. The degree of acetylation (DA) values determined by the 1H NMR analysis increased with the higher reaction temperatures. The thermal gravimetric analysis (TGA) indicated the enhancement of thermal stability by acetylation. Furthermore, the TGA as well as differential scanning calorimetric (DSC) analysis of the products suggested the presence of the highly and less acetylated segments in a xanthan gum chain. The DSC profile of the product with the high DA value also exhibited a small endothermic peak, which might potentially be ascribed to the melting temperature.

Background Isoflurane and sevoflurane protect lungs with ischemia–reperfusion (IR) injury. We examined the influence of desflurane on IR lung injury using isolated rabbit lungs perfused with a physiological salt solution. Methods The isolated lungs were divided into three groups: IR, desflurane-treated ischemia–reperfusion (DES-IR), and ventilation/perfusion-continued control (Cont) groups (n = 6 per group). In the DES-IR group, inhalation of desflurane at 1 minimum alveolar concentration (MAC) was conducted in a stable 30-min phase. In the IR and DES-IR groups, ventilation/perfusion was stopped for 75 min after the stable phase. Subsequently, they were resumed. Each lung was placed on a balance, and weighed. Weight changes were measured serially throughout this experiment. The coefficient of filtration (Kfc) was determined immediately before ischemia and 60 min after reperfusion. Furthermore, bronchoalveolar lavage fluid (BALF) was collected from the right bronchus at the completion of the experiment. After the completion of the experiment, the left lung was dried, and the lung wet-to-dry weight ratio (W/D) was calculated. Results The Kfc values at 60 min after perfusion were 0.40 ± 0.13 ml/min/mmHg/100 g in the DES-IR group, 0.26 ± 0.07 ml/min/mmHg/100 g in the IR group, and 0.22 ± 0.08 (mean ± SD) ml/mmHg/100 g in the Cont group. In the DES-IR group, the Kfc at 60 min after the start of reperfusion was significantly higher than in the other groups. In the DES-IR group, W/D was significantly higher than in the Cont group. In the DES-IR group, the BALF concentrations of nitric oxide metabolites were significantly higher than in the other groups. In the DES-IR group, the total amount of vascular endothelial growth factor in BALF was significantly higher than in the Cont group. Conclusions The pre-inhalation of desflurane at 1 MAC exacerbates pulmonary IR injury in isolated/perfused rabbit lungs.

Cyclin-dependent kinase (Cdk)5 is a proline-directed Ser/Thr protein kinase that functions mainly in neurons and is activated by binding to a regulatory subunit, p35 or p39. Kinase activity is mainly determined by the amount of p35 available, which is controlled by a balance between synthesis and degradation. Kinase activity is also regulated by Cdk5 phosphorylation, but the activity of phosphorylated Cdk5 is in contrast to that of cycling Cdks. Cdk5 is a versatile protein kinase that regulates multiple neuronal activities including neuronal migration and synaptic signaling. Further, Cdk5 plays a role in both survival and death of neurons. Long-term inactivation of Cdk5 triggers cell death, and the survival activity of Cdk5 is apparent when neurons suffer from stress. In contrast, hyper-activation of Cdk5 by p25 promotes cell death, probably by reactivating cell-cycle machinery in the nucleus. The pro-death activity is suppressed by membrane association of Cdk5 via myristoylation of p35. Appropriate activity, localization, and regulation of Cdk5 may be critical for long-term survival of neurons, which is more than 80 years in the case of humans.

The dysfunction of proteasomes and mitochondria has been implicated in the pathogenesis of Parkinson disease. However, the mechanism by which this dysfunction causes neuronal cell death is unknown. We studied the role of cyclin-dependent kinase 5 (Cdk5)-p35 in the neuronal cell death induced by 1-methyl-4-phenylpyrinidinium ion (MPP+), which has been used as an in vitro model of Parkinson disease. When cultured neurons were treated with 100 microM MPP+, p35 was degraded by proteasomes at 3 h, much earlier than the neurons underwent cell death at 12-24 h. The degradation of p35 was accompanied by the down-regulation of Cdk5 activity. We looked for the primary target of MPP+ that triggered the proteasome-mediated degradation of p35. MPP+ treatment for 3 h induced the fragmentation of the mitochondria, reduced complex I activity of the respiratory chain without affecting ATP levels, and impaired the mitochondrial import system. The dysfunction of the mitochondrial import system is suggested to up-regulate proteasome activity, leading to the ubiquitin-independent degradation of p35. The overexpression of p35 attenuated MPP+-induced neuronal cell death. In contrast, depletion of p35 with short hairpin RNA not only induced cell death but also sensitized to MPP+ treatment. These results indicate that a brief MPP+ treatment triggers the delayed neuronal cell death by the down-regulation of Cdk5 activity via mitochondrial dysfunction-induced up-regulation of proteasome activity. We propose a role for Cdk5-p35 as a survival factor in countering MPP+-induced neuronal cell death.

Prion diseases are fatal neurodegenerative disorders, and the conformational conversion of normal cellular prion protein (PrP(C)) into its pathogenic, amyloidogenic isoform (PrP(Sc)) is the essential event in the pathogenesis of these diseases. Lactoferrin (LF) is a cationic iron-binding glycoprotein belonging to the transferrin (TF) family, which accumulates in the amyloid deposits in the brain in neurodegenerative disorders, such as Alzheimer's disease and Pick's disease. In the present study, we have examined the effects of LF on PrP(Sc) formation by using cell culture models. Bovine LF inhibited PrP(Sc) accumulation in scrapie-infected cells in a time- and dose-dependent manner, whereas TF was not inhibitory. Bioassays of LF-treated cells demonstrated prolonged incubation periods compared with non-treated cells indicating a reduction of prion infectivity. LF mediated the cell surface retention of PrP(C) by diminishing its internalization and was capable of interacting with PrP(C) in addition to PrP(Sc). Furthermore, LF partially inhibited the formation of protease-resistant PrP as determined by the protein misfolding cyclic amplification assay. Our results suggest that LF has multifunctional antiprion activities.

Pleomorphic adenoma is the most frequent salivary gland tumor. Pleomorphic adenoma occurring from the allopatry salivary gland is particularly rare. We experienced a 28-year-old woman with pleomorphic adenoma thought to have occurred from the allopatry salivary gland on the ipsilateral side chin lower than the left parotoid pleomorphic adenoma for which one had undergone surgery eight years previously. Imaging (CT, MR etc) demonstrated an oval-shaped tumor located in the left lower chin without evidence of connection with the left submandibular gland and left parotid gland. Surgically, the tumor was completely removed and pathologically diagnosed as pleomorphic adenoma. Finally, we concluded that it was pleomorphic adenoma arising from the allopatry salivary gland. However, it could not be completely excluded that it was metastasis from the parotoid pleomorphic adenoma on the ipsilateral side from eight years previously.

下咽頭癌T3，T4症例に対し当科で施行している化学放射線同時併用療法の治療効果を検討した。合併症のない症例に対し，放射線治療は61.2～72Gy，化学療法はドセタキセル（DOC），シスプラチン（CDDP），5-FUの3剤（TPF療法）またはCDDP，5-FU，メソトレキセート（MTX），ロイコボリン（LV）の4剤（CFML療法）を用いた。24症例が対象となり， T3：8例，T4：16例，N0：1例，N2b：9例，N2c：10例，N3：4例であった。一次治療後，原発巣でのCR率はT3で75％，T4で56％，頸部リンパ節でのCR率はN2bで33％，N2cで20％，N3で25％であった。生存率はT3とT4との間に有意差を認めたが，N分類では有意差を認めなかった。進行期であるT3，T4症例ではほとんどが複数のリンパ節転移を有するため，原発巣の進展範囲によって治療効果に有意差を生じたものと考察された。

Objective: The eosinophilic-otitis-media have characteristic pooling of viscous liquids and granulation tissues in the tympanic cavity. The basic treatment is removal of granulation tissue allowed by an eardrum incision, steroid injection into the tympanic cavity. Therefore, we made eardrum incisions and removed the fluid accumulation and granulation tissues. However, this treatment may induce postoperative eardrum perforation. Therefore, to avoid eardrum perforation after treatment, we injected steroid liquids into tympanic cavity using a 23G fine needle without making an eardrum incision or inserting a tube into the tympanic cavity. Then, we examined the treatment outcomes.Materials and Methods: We diagnosed 17 cases, 33 ears (male: 2 cases 4 ears, female: 15 cases, 29 ears) (average age: 51.4 years old) as having eosinophilic-otitis-media without perforations based on standard diagnostic criteria. Then, we retrospectively investigated the hearing levels and eardrum perforation after treatment by dividing two groups. One group was treated without making eardrum incisions using a 23G fine needle to inject steroid solution, into the tympanic cavity along with oral steroid administration. The other group was treated by making an eardrum incision or inserting a tube, then, injecting steroid solution into the tympanic cavity along with oral steroid administration.Result: The group undergoing eardrum incisions had an eardrum perforation rate of 76.4% (13/17 ears) after treatment. However, the group without eardrum incisions had an eardrum perforation rate of 25.0% (4/16 ears) after treatment. Regarding hearing improvement, there was not a meaningful hearing improvement after treatment in the group receiving eardrum incisions. However, there was a meaningful postoperative hearing improvement of 500-4000Hz in the group without eardrum incisions.Conclusion: We found that treating eosinophilic-otitis-media without eardrum incisions by using a fine needle to inject steroid solution into the tympanic cavity along with oral steroid administration significantly diminished the rate of postoperative perforations.

Surface-initiated graft atom transfer radical polymerization (ATRP) of methyl methacrylate (MMA) from self-assembled chitin nanofibers (CNFs) was performed under dispersion conditions. Self-assembled CNFs were initially prepared by regeneration from a chitin ion gel with 1-allyl-3-methylimidazolium bromide using methanol, the product was then converted into the chitin nanofiber macroinitiator by reaction with α-bromoisobutyryl bromide in a dispersion containing N,N-dimethylformamide. Surface-initiated graft ATRP of MMA from the initiating sites on the CNFs was subsequently carried out under dispersion conditions, followed by filtration to obtain the CNF-graft-polyMMA film. Analysis of the product confirmed the occurrence of the graft ATRP on the surface of the CNFs.

Soil microbial communities have great potential for bioremediation of recalcitrant aromatic compounds. However, it is unclear which taxa and genes in the communities, and how they contribute to the bioremediation in the polluted soils. To get clues about this fundamental question here, time-course (up to 24 weeks) metagenomic analysis of microbial community in a closed soil microcosm artificially polluted with four aromatic compounds, including phenanthrene, was conducted to investigate the changes in the community structures and gene pools. The pollution led to drastic changes in the community structures and the gene sets for pollutant degradation. Complete degradation of phenanthrene was strongly suggested to occur by the syntrophic metabolism by Mycobacterium and the most proliferating genus, Burkholderia. The community structure at Week 24 (-12 weeks after disappearance of the pollutants) returned to the structure similar to that before pollution. Our time-course metagenomic analysis of phage genes strongly suggested the involvement of the 'kill-the-winner' phenomenon (i.e. phage predation of Burkholderia cells) for the returning of the microbial community structure. The pollution resulted in a decrease in taxonomic diversity and a drastic increase in diversity of gene pools in the communities, showing the functional redundancy and robustness of the communities against chemical disturbance.

Olprinone is an inotropic agent that inhibits phosphodiesterase (PDE) III and causes vasodilation. Olprinone has been shown to be less proarrhythmic and possibly affect expression of functional Kv1.5 channels that confer the ultra-rapid delayed-rectifier K+ channel current (IKur) responsible for action potential repolarization. To reveal involvement of Kv1.5 channels in the less arrhythmic effect of olprinone, we examined effects of the agent on the stability of Kv1.5 channel proteins expressed in COS7 cells. Olprinone at 30-1000 nM increased the protein level of Kv1.5 channels in a concentration-dependent manner. Chase experiments showed that olprinone delayed degradation of Kv1.5 channels. Olprinone increased the immunofluorescent signal of Kv1.5 channels in the endoplasmic reticulum (ER) and Golgi apparatus as well as on the cell surface. Kv1.5-mediated membrane currents, measured as 4-aminopyridine-sensitive currents, were increased by olprinone without changes in their activation kinetics. A protein transporter inhibitor, colchicine, abolished the olprinone-induced increase of Kv.1.5-mediated currents. The action of olprinone was inhibited by 4-aminopyridine, and was not mimicked by the application of 8-Bromo-cAMP. Taken together, we conclude that olprinone stabilizes Kv1.5 proteins at the ER through an action as a chemical chaperone, and thereby increases the density of Kv1.5 channels on the cell membrane. The enhancement of Kv1.5 currents could underlie less arrhythmogenicity of olprinone.

Sphingobium japonicum (formerly Sphingomonas paucimobilis) UT26 utilizes the important insecticide gamma-hexachlorocyclohexane as a sole source of carbon and energy. In previous studies, we isolated and characterized six structural genes (linA to linF) and one regulatory gene (linR) of UT26 for the degradation of gamma-hexachlorocyclohexane to beta-ketoadipate. Our analysis in this study indicated that the UT26 genome consists of three large circular replicons of 3.6 Mb, 670 kb, and 185 kb. The 3.6 Mb and the 670 kb replicons had one and two copies, respectively, of the 16S ribosomal RNA gene, and these replicons were designated as chromosomes (Chr) I and II, respectively. Chr I was indicated to be a main chromosome carrying the dnaA gene. The first three lin genes, linA to linC, for conversion of gamma-hexachlorocyclohexane to 2,5-dichlorohydroquinone, were dispersed on Chr I. The 185 kb plasmid, pCHQ1, carried the linRED operon for the conversion of 2,5-dichlorohydroquinone to maleylacetate and was conjugatively transferred to another sphingomonad strain. The linF gene encoding maleylacetate reductase was located on Chr II. These results indicated that the genes for the complete gamma-hexachlorocyclohexane degradation are dispersed on the three large replicons of UT26.