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Abstract Background Biomarkers that can accurately predict the efficacy of immune checkpoint inhibitors (ICIs) against programmed death 1 (PD-1) ligand in cancer immunotherapy are urgently needed. We have previously reported a novel formula that predicts the response to treatment with second-line nivolumab with high sensitivity and specificity in patients with non-small cell lung cancer (NSCLC) previously treated with chemotherapy. The formula was based on the percentages of CD62LlowCD4+ T cells (effector T cells; %Teff) and CD4+CD25+FOXP3+ T cells (regulatory T cells; %Treg) in the peripheral blood before treatment estimated using the peripheral blood mononuclear cell (PBMC) method. Here, we investigated the applicability of the formula (K-index) to predict the response to treatment with another ICI to expand its clinical applicability. Furthermore, we developed a simpler assay method based on whole blood (WB) samples to overcome the limitations of the PBMC method, such as technical difficulties, in obtaining the K-index. Methods The K-index was evaluated using the PBMC method in 59 patients with NSCLC who received first-line pembrolizumab treatment. We also assessed the K-index using the WB method and estimated the correlation between the measurements obtained using both methods in 76 patients with lung cancer. Results This formula consistently predicted the response to first-line pembrolizumab therapy in patients with NSCLC. The WB method correlated well with the PBMC method to obtain %Teff, %Treg, and the formula value. The WB method showed high repeatability (coefficient of variation,

Abstract This study quantitatively evaluated the typhoon‐induced sea surface temperature (SST) cooling caused by typhoons Faxai (2019) and Hagibis (2019) using a high‐resolution ocean model and the cooling parameter (Co). Faxai and Hagibis both passed over the ocean south of the eastern part of the Japanese main island, but the associated average SST decreases differed. Faxai produced a decrease of less than 2°C, whereas Hagibis produced a decrease of more than 2°C. The average Co value was 1.6 for Faxai and 3.6 for Hagibis, indicating that SST was more easily cooled by Hagibis than by Faxai, consistent with the observations. The impact of ocean conditions on the typhoon‐induced SST cooling by Hagibis was 2.6 times larger than the impact by Faxai, indicating that the ocean before Hagibis passes is less hard to cool ocean than Faxai. In short, it is important for ocean cooling not only ocean conditions but also typhoon characteristics because in fact, Hagibis cooled the ocean more than Faxai. In addition, the impact of Hagibis's characteristics on the typhoon‐induced SST cooling was 4.8 times larger than the impact of Faxai's characteristics. Thus, SST was more likely to cool by typhoon characteristics in the case of Hagibis. In particular, among Hagibis's characteristics, typhoon size in the horizontal direction had the most efficient effect on SST cooling. Although Co does not consider the effects of the advection of ocean water, we suggest that Co is a practical indicator for estimating SST cooling caused by a typhoon and comparing factors of typhoon‐induced SST cooling in multiple cases.

Abstract This study analyzed the observed data of precipitation and PM2.5 concentration collected over two and a half years at Tsujido, Fujisawa City, to examine a quantitative relationship between the two quantities. It was observed that the PM2.5 concentration decreased as precipitation increased. A composite analysis revealed that the PM2.5 concentration on average decreases by 20.99% 1 h after the onset of precipitation as compared to that 1 h before, which is statistically significant based on a paired t‐test. The PM2.5 concentrations decreased over 8 h period from 5 h (−5 h) before to 3 h (+3 h) after the onset of precipitation, with a particularly dramatic decrease at the onset of precipitation. Analysis of PM2.5 scavenging rates by precipitation intensity showed that the scavenging rate was 2.03% for light precipitation events, 28.15% for moderate precipitation events, and 26.75% for heavy precipitation events, indicating that the scavenging rate increased with moderate precipitation intensity and above. It is suggested that the deposition processes of rainout and washout are effective in reducing PM2.5 concentration with precipitation.

This is a prospective study that has been done to assess the lumbar sagittal alignment of patients positioned prone over an adjustable hinged carbon fiber operating table. The following three variations of table position have been considered: neutral, 20° convex, and 20° concave bending. A total of 33 patients who underwent lumbar disc herniation and lumbar canal stenosis surgery were enrolled. Patients who presented with spinal deformity akin to lumbar scoliosis or spondylolisthesis were excluded. For the surgical procedure, following the induction of endotracheal general anesthesia, patients were positioned prone on the adjustable hinged carbon fiber operating table. Radiographs of the lateral view of the lumbar spine were acquired for the three table positions, i.e., neutral, 20° convex, and 20° concave. The lumbar lordosis was measured on radiographs. The lumbar lordosis presented divergently in all three variations of the table (p < 0.01): 45.2 ± 11.0° in neutral; 52.0 ± 10.7° in 20° lumbar extension bending; and 35.9 ± 10.8° in 20° lumbar flexion bending. The efficacy of table bending was 46.5% in convex and 34.0% in concave bending. In conclusion, the lumbar lordosis in three different table positions were 35.9° in 20° convex bending, 45.2° in neutral, and 52.0° in concave bending. The efficacy of table bending was 46.5% in convex and 34.0% in concave bending.

Abstract The impacts of aerosols on the charge distribution of hydrometeors and lightning flash density in a tropical cyclone (TC) were investigated using a meteorological model coupled with an explicit lightning model. The meteorological model successfully simulated the tripole structure of charge density distribution in a TC, as reported by previous studies. The impacts of aerosols were investigated through a sensitivity experiment with changing the aerosol number concentration. The tripole structure became unclear with increasing aerosol number concentrations. The positive charge distribution located in the lower layer was not seen, and raindrops with negative charge distribution reached the surface. As a result, the vertical structure of the charge density was dipolar in the polluted case. As the tripole structure shifted to dipole, the magnitude of the electric field tended to be large, and the flash number was large. By contrast, in the pristine case, the tripole structure was dominant, and the flash number was much smaller than in the polluted case.

Abstract This article reviews the major outcomes of a 5-year (2011–2016) project using the K computer to perform global numerical atmospheric simulations based on the non-hydrostatic icosahedral atmospheric model (NICAM). The K computer was made available to the public in September 2012 and was used as a primary resource for Japan’s Strategic Programs for Innovative Research (SPIRE), an initiative to investigate five strategic research areas; the NICAM project fell under the research area of climate and weather simulation sciences. Combining NICAM with high-performance computing has created new opportunities in three areas of research: (1) higher resolution global simulations that produce more realistic representations of convective systems, (2) multi-member ensemble simulations that are able to perform extended-range forecasts 10–30 days in advance, and (3) multi-decadal simulations for climatology and variability. Before the K computer era, NICAM was used to demonstrate realistic simulations of intra-seasonal oscillations including the Madden-Julian oscillation (MJO), merely as a case study approach. Thanks to the big leap in computational performance of the K computer, we could greatly increase the number of cases of MJO events for numerical simulations, in addition to integrating time and horizontal resolution. We conclude that the high-resolution global non-hydrostatic model, as used in this five-year project, improves the ability to forecast intra-seasonal oscillations and associated tropical cyclogenesis compared with that of the relatively coarser operational models currently in use. The impacts of the sub-kilometer resolution simulation and the multi-decadal simulations using NICAM are also reviewed.

Transmembrane protein 168 (TMEM168) comprises 697 amino acid residues, including some putative transmembrane domains. It is reported that TMEM168 controls methamphetamine (METH) dependence in the nucleus accumbens (NAc) of mice. Moreover, a strong link between METH dependence-induced adaptive changes in the brain and mood disorders has been evaluated. In the present study, we investigated the effects of accumbal TMEM168 in a battery of behavioral paradigms. The adeno-associated virus (AAV) Tmem168 vector was injected into the NAc of C57BL/6J mice (NAc-TMEM mice). Subsequently, the accumbal TMEM168 mRNA was increased approximately by seven-fold when compared with the NAc-Mock mice (controls). The NAc-TMEM mice reported no change in the locomotor activity, cognitive ability, social interaction, and depression-like behaviors; however, TMEM168 overexpression enhanced anxiety in the elevated-plus maze and light/dark box test. The increased anxiety was reversed by pretreatment with the antianxiety drug diazepam (0.3 mg/kg i.p.). Moreover, the NAc-TMEM mice exhibited decreased prepulse inhibition (PPI) in the startle response test, and the induced schizophrenia-like behavior was reversed by pretreatment with the antipsychotic drug risperidone (0.01 mg/kg i.p.). Furthermore, accumbal TMEM168 overexpression decreased the basal levels of extracellular GABA in the NAc and the high K+ (100 mM)-stimulated GABA elevation; however, the total contents of GABA in the NAc remained unaffected. These results suggest that the TMEM168-regulated GABAergic neuronal system in the NAc might become a novel target while studying the etiology of anxiety and sensorimotor gating deficits.

Shati/Nat8L significantly increased in the nucleus accumbens (NAc) of mice after repeated methamphetamine (METH) treatment. We reported that Shati/Nat8L overexpression in mouse NAc attenuated METH-induced hyperlocomotion, locomotor sensitization, and conditioned place preference. We recently found that Shati/Nat8L overexpression in NAc regulates the dopaminergic neuronal system via the activation of group II mGluRs by elevated N-acetylaspartylglutamate following N-acetylaspartate increase due to the overexpression. These findings suggest that Shati/Nat8L suppresses METH-induced responses. However, the mechanism by which METH increases the Shati/Nat8L mRNA expression in NAc is unclear. To investigate the regulatory mechanism of Shati/Nat8L mRNA expression, we performed a mouse Shati/Nat8L luciferase assay using PC12 cells. Next, we investigated the response of METH to Shati/Nat8L expression and CREB activity using mouse brain slices of NAc, METH administration to mice, and western blotting for CREB activity of specific dopamine receptor signals in vivo and ex vivo. We found that METH activates CREB binding to the Shati/Nat8L promoter to induce the Shati/Nat8L mRNA expression. Furthermore, the dopamine D1 receptor antagonist SCH23390, but not the dopamine D2 receptor antagonist sulpiride, inhibited the upregulation of Shati/Nat8L and CREB activities in the mouse NAc slices. Thus, the administration of the dopamine D1 receptor agonist SKF38393 increased the Shati/Nat8L mRNA expression in mouse NAc. These results showed that the Shati/Nat8L mRNA was increased by METH-induced CREB pathway via dopamine D1 receptor signaling in mouse NAc. These findings may contribute to development of a clinical tool for METH addiction.

Abstract.: A devastating psychiatric disorder, schizophrenia is characterized by three major symptoms, positive and negative symptoms and cognitive deficit. Almost all current therapeutic drugs for schizophrenia have efficacy for positive symptoms, and weak efficacy for negative and cognitive deficit. In particular, social withdrawal, diminished motivation, and anhedonia as the depressive aspects of negative symptoms are resistant to the treatment of antipsychotic drugs. Therefore, there is a need for development of new therapeutic drugs for negative symptoms of schizophrenia, and it is necessary to have comprehensive animal models to understand the neurobiological foundations of their symptoms. In this review, we represent the behavioral phenotypes in available animal models of schizophrenia for drug discovery, focusing on the depressive aspects of negative symptoms. We mention here animal models based on the pathology and epidemiology of schizophrenia, e.g., the pharmacological, neurodevelopmental, genetic, and gene-environment combination models. The animal models of schizophrenia are developed by various approaches and are assessed, but there are few models demonstrating negative symptoms with sensitivities to available therapeutic drugs. The development of comprehensive animal model reflecting negative symptoms and of novel compounds that can remedy them provide certain insight into the neurobiological process of schizophrenia and also point the way to a new therapeutic strategy. Keywords:: schizophrenia, social withdrawal, diminished motivation, anhedonia, animal models

The number of patients with schizophrenia has increased over the past decade. Previously, many studies have been performed to establish its diagnostic criteria, prophylactic methods, and effective therapies. In this study, we analyzed whether the ratios of DNA methylation in CpG islands of the Shati/Nat8l is decreased in model mice of schizophrenia-like phenotype using genomic DNA collected from brain regions and peripheral blood, since the mouse model of schizophrenia-like phenotype, mice treated repeatedly with methamphetamine showed increase of Shati/Nat8l mRNA expression in our previous experiment. The ratios of Shati/Nat8l CpG island methylation were significantly decreased in both the nucleus accumbens and the peripheral blood of model mice compared with those of control mice. We also investigated Shati/Nat8l methylation in the blood of patients with schizophrenia. We found that Shati/Nat8l CpG island methylation ratios were lower in the patients with schizophrenia than in the healthy controls, which is consistent with our findings in the mice model. To our knowledge, this is the first study to show similar alterations in methylation status of a particular genomic DNA site in both the brain and peripheral blood of mice. Furthermore, the same phenomenon was observed in corresponding human genomic sequences of the DNA extracted from the peripheral blood of patients with schizophrenia. Based on our findings, DNA methylation profiles of the CpG island of Shati/Nat8l might be a diagnostic biomarker of schizophrenia.

The single nucleotide polymorphism (SNP) rs13438494 in intron 24 of PCLO was significantly associated with bipolar disorder in a meta-analysis of genome-wide association studies. In this study, we performed functional minigene analysis and bioinformatics prediction of splicing regulatory sequences to characterize the deep intronic SNP rs13438494. We constructed minigenes with A and C alleles containing exon 24, intron 24, and exon 25 of PCLO to assess the genetic effect of rs13438494 on splicing. We found that the C allele of rs13438494 reduces the splicing efficiency of the PCLO minigene. In addition, prediction analysis of enhancer/silencer motifs using the Human Splice Finder web tool indicated that rs13438494 induces the abrogation or creation of such binding sites. Our results indicate that rs13438494 alters splicing efficiency by creating or disrupting a splicing motif, which functions by binding of splicing regulatory proteins, and may ultimately result in bipolar disorder in affected people.

In 2019, serious disasters were caused by local strong winds associated with Typhoon Faxai and wide torrential rains with Typhoon Hagibis. Although both tropical cyclones (TCs) followed similar tracks and underwent extratropical transition after the recurvature (ETR), their storm sizes and structures were distinct: Faxai was a small axisymmetric TC, whereas Hagibis was a large asymmetric TC. The purpose of this study is to clarify the effect of storm size on a TC that undergoes ET and its associated synoptic environment. Hagibis causes a larger amount of precipitation more widely than Faxai. A large amount of diabatic heating closely associated with the precipitation leads to low potential vorticity (PV) production downstream of Hagibis in the upper troposphere and the enhancement of the ridge. By contrast, the diabatic heating is relatively small, and the production of low PV area is indistinct downstream of Faxai. Besides the case studies, large (LA) and small (SM) TCs that undergo ETR (LA-ETR and SM-ETR TCs, respectively) in the western North Pacific from 2016 to 2020 are statistically compared using cyclone phase space and composite analyses with the best track and Japanese 55-year Reanalysis datasets. As observed in the case studies, the LA-ETR TCs are characterized by a larger amount of diabatic heating and a more enhancement of the downstream ridge than the SM-ETR TCs. The LA-ETR TCs change into asymmetric structures more drastically than the SM-ETR TCs while moving northward along the westerly jet with increasing the amplitude of the north-south meander. By contrast, the amplitude of the north-south meander of the westerly jet does not increase around the SM-ETR TCs. Therefore, the larger the storm size is, the larger the amplitude of the north-south meander of the westerly jet is, resulting in a more drastic asymmetric structural change of the TC. [ABSTRACT FROM AUTHOR]

A numerical simulation of Typhoon Faxai (1915), which made landfall with a central pressure of 960 hPa in the Kanto region of Japan, was conducted using a nonhydrostatic numerical model with a 1-km grid spacing. Faxai sustained a symmetric structure until the landfall and caused severe damage due to strong winds. The simulation successfully simulated the realistic track and intensity of Faxai for 48 h around landfall. The simulated intensity was strong until the time of landfall, and the spatial size of the vortex was small. The structure of the simulated Faxai, identified as having an axisymmetric flow field and eyewall, was similar to that of a welldeveloped tropical cyclone (TC) in the tropics. Around the TC center, the surface latent heat flux was over 300 W m-2 until landfall, and the vertical wind shear was less than 9 m s-1 between the 1.5- and 12.0-km altitudes, which is relatively weak at midlatitudes. The maximum potential intensity (MPI) was calculated using environmental parameters around the simulated TC. The simulated and best track TC intensities exceeded the MPI for approximately 12 h before landfall, that is, the TC was in a superintense state. The superintensity was mainly caused by the presence of supergradient wind, which, in turn, resulted from the strong intensity and axisymmetric structure of the typhoon. The simulated TC satisfies the assumptions for the formulation of the MPI during the quasi-steady state, except for the gradient wind balance, implying that the structure of a TC is similar to that of a developed TC in the tropics. The present analyses suggest that the strong intensity of Faxai results from favorable environmental conditions and vortex structure. [ABSTRACT FROM AUTHOR]

The dependence of lightning frequency on the life cycle of an idealized tropical cyclone (TC) was investigated using a three-dimensional meteorological model coupled with an explicit lightning model. To investigate this dependence, an idealized numerical simulation covering the initial state to the steady state (SS) of an idealized TC was conducted. The simulation was consistent with the temporal evolution of lightning frequency reported by previous observational studies. Our analyses showed that the dependence originates from changes in the types of convective cloud with lightning over the life cycle of the TC. Before rapid intensification (RI) and in the early stage of RI, convective cloud cells that form under high-convective available potential energy (CAPE) conditions are the main contributors to lightning. As the TC reaches the late stage of RI and approaches SS, the secondary circulation becomes prominent and convective clouds in the eyewall region alongside the secondary circulation gradually become the main contributors to the lightning. In the convective cloud cells formed under high-CAPE conditions, upward velocity is strong and large charge density is provided through noninductive charge separation induced by graupel collisions. This large charge density frequently induces lightning in the clouds. On the other hand, the vertical velocity in the eyewall is weak, and it tends to contribute to lightning only when the TC reaches the mature stage. Our analyses imply that the maximum lightning frequency that occurs before the maximum intensity of a TC corresponds to the stage of a TC's life cycle in which convective cloud cells are generated most frequently and moisten the upper troposphere. [ABSTRACT FROM AUTHOR]

The impacts of the number density of cloud condensation nuclei (CCN) and other thermodynamic quantities on moist Rayleigh convection were examined. A numerical model, consisting of a simple two-dimensional equation for Boussinesq air and a sophisticated double moment microphysics scheme, was developed. The impact of the number of CCN is most prominent in the initially formed convection, whereas the convection in the quasi-steady state does not significantly depend on the number of CCN. It is suggested that the former convection is driven by a mechanism without a background circulation, such as parcel theory. In contrast, the latter convection appears to be driven by the statically unstable background layer. Incorporating the cloud microphysics reduces the integrated kinetic energy and number of convective cells (increases the distance between the cells), with some exceptions, which are consistent with previous studies. These features are not largely sensitive to the number of CCN. It is shown in this study that the reduction in kinetic energy is mainly due to condensation (evaporation) in the upper (lower) layer, which tends to stabilize the fluid. The ensemble simulation shows that the sensitivity of the moist processes to changes in the temperature at the bottom boundary, temperature lapse rate, water vapor mixing ratio, and CCN is qualitatively similar to that in the control simulation. The impact becomes strong with increasing temperature lapse rate. The number of convective cells in a domain decreases with the degree of supersaturation or an increase in the domain-integrated condensate. [ABSTRACT FROM AUTHOR]

Background: Several clinical studies have suggested that N-acetylaspartate and N-acetylaspartylglutamate levels in the human brain are associated with various psychiatric disorders, including major depressive disorder. We have previously identified Shati/Nat8l, an N-acetyltransferase, in the brain using an animal model of psychosis. Shati/Nat8l synthesizes N-acetylaspartate from L-aspartate and acetyl-coenzyme A. Further, N-acetylaspartate is converted into N-acetylaspartylglutamate, a neurotransmitter for metabotropic glutamate receptor 3. Methods: Because Shati/Nat8l mRNA levels were increased in the dorsal striatum of mice following the exposure to forced swimming stress, Shati/Nat8l was overexpressed in mice by the microinjection of adeno-associated virus vectors containing Shati/Nat8l gene into the dorsal striatum (dS-Shati/Nat8l mice). The dS-Shati/Nat8l mice were further assessed using behavioral and neurochemical tests. Results: The dS-Shati/Nat8l mice exhibited behavioral despair in the forced swimming and tail suspension tests and social withdrawal in the 3-chamber social interaction test. These depression-like behaviors were attenuated by the administration of a metabotropic glutamate receptor 2/3 antagonist and a selective serotonin reuptake inhibitor. Furthermore, the metabolism of N-acetylaspartate to N-acetylaspartylglutamate was decreased in the dorsal striatum of the dS-Shati/Nat8l mice. This finding corresponded with the increased expression of glutamate carboxypeptidase II, an enzyme that metabolizes N-acetylaspartylglutamate present in the extracellular space. Extracellular serotonin levels were lower in the dorsal striatum of the dS-Shati/Nat8l and normal mice that were repeatedly administered a selective glutamate carboxypeptidase II inhibitor. Conclusions: Our findings indicate that the striatal expression of N-acetylaspartate synthetase Shati/Nat8l plays a role in major depressive disorder via the metabotropic glutamate receptor 3-mediated functional control of the serotonergic neuronal system. [ABSTRACT FROM AUTHOR]

The relationship of tropical cyclone (TC) future intensity change to current intensity and current axisymmetricity deduced from hourly Global Satellite Mapping of Precipitation (GSMaP) data was investigated. Axisymmetricity is a metric that correlates positively with the magnitude of the axisymmetric component of the rainfall rate and negatively with the magnitude of the asymmetric component. The samples used were all of the TCs that existed in the western North Pacific basin during the years 2000-15. The results showed that, during the development stage, the intensification rate at the current time, and 6 and 12 h after the current time was strongly related to both the current intensity and axisymmetricity. On average, the higher the axisymmetricity, the larger the intensity change in the next 24 h for TCs with a current central pressure (maximum sustained wind) between 945 and 995 hPa (85 and 40 kt). The mean value of the axisymmetricity for TCs experiencing rapid intensification (RI) was much higher than that for non-RI TCs for current intensities of 960-990 hPa. The new observational evidence for the intensification process presented here is consistent with the findings of previous theoretical studies emphasizing the role of the axisymmetric component of diabatic heating. [ABSTRACT FROM AUTHOR]

The environmental conditions for tropical cyclone genesis are examined by numerical experiment. We focus on the case of a non-developing disturbance showed features for tropical cyclone genesis in the Pacific Area Long-term Atmospheric observation for Understanding climate change in 2010 (PALAU2010) observation campaign over the western North Pacific. We clarify the importance of the presence of abundant moisture around the disturbance for continuous convection and demonstrate that the collocation of a mid-level vortex and a low-level vortex, i.e., the persistence of an upright structure of vortices, is important in tropical cyclone genesis. We conduct two numerical experiments using the Weather Research and Forecasting Model Advanced Research WRF model in double nested domains with a horizontal grid space of 27 km and 9 km for the outer domain and the inner domain, respectively. The first experiment is based on reanalysis data (a control experiment) and the second includes increased water vapor content over the northwestern dry area of the disturbance. In the control experiment, the disturbance did not develop into a tropical cyclone in spite of the existence of the mid-level and low-level vortices. In contrast, the sensitivity experiment shows that a tropical cyclone was formed from the disturbance with increased water vapor content. The presence of persistent upright vortices was supported by continuous convection until the genesis of the tropical cyclone. [ABSTRACT FROM AUTHOR]

We examine how cloud cover is determined in shallow-cloud areas by using large-eddy simulation with an extremely wide domain, which covers the transition phase from cumulus-understratocumulus to shallow-cumulus regimes. The relationship between two distances is critical to cloud cover. One characteristic distance is the horizontal distance between cumulus clouds, and the other is the broadening distance of anvil-like stratiform cloud at the top of the boundary layer. High cloud cover occurs with a long distance of broadening and short distances between cumuli. In contrast, low cloud cover appears with a short distance of broadening and a long distance between cumuli. The contrast of the two distances is rooted in aerosol amount and the strength of the surface heat flux. The relationship between these two distances can be applied to estimating the cloud cover below sharp inversions. [ABSTRACT FROM AUTHOR]

A Lewis acid-catalyzed [3+1+1] cycloaddition reaction between aliphatic isocyanides and azomethine ylides generated in situ from aziridines, leading to pyrrolidine derivatives, has been developed. This reaction proceeds smoothly under mild conditions and can also be modified by employing aromatic isocyanides to generate four-membered heterocycles, azetidines, through a [3+1] cycloaddition reaction. [ABSTRACT FROM AUTHOR]

The mechanisms of NO inhibition of CaMK [Ca2+/CaM (calmodulin)-dependent protein kinase] II activity were studied. In rat pituitary tumour GH3 cells, TRH [thyrotrophin (TSH)-releasing hormone]-stimulated phosphorylation of nNOS [neuronal NOS (NO synthase)] at Ser847 was sensitive to an inhibitor of CaMKs, KN-93, and was enhanced by inhibition of nNOS with 7NI (7-nitroindazole). Enzyme activity of CaMKII following in situ treatment with 7NI was also increased. The in vitro activity of CaMKII was inhibited by co-incubation either with nNOS and L-arginine or with NO donors SNAP (S-nitroso-N-acetyl-DL-penicillamine) and DEA-NONOate [diethylamine-NONOate (diazeniumdiolate)]. Once inhibited by these treatments, CaMKII was observed to undergo full reactivation on the addition of a reducing reagent, DTT (dithiothreitol). In transfected cells expressing CaMKII and nNOS, treatment with the calcium ionophore A23187 further revealed nNOS phosphorylation at Ser847, which was enhanced by 7NI and CaMKII S-nitrosylation. Mutated CaMKII (C6A), in which Cys6 was substituted with an alanine residue, was refractory to 7NI-induced enhancement of nNOS phosphorylation or to CaMKII S-nitrosylation. Furthermore, we could identify Cys6 as a direct target for S-nitrosylation of CaMKII using MS. In addition, treatment with glutamate caused an increase in CaMKII S-nitrosylation in rat hippocampal slices. This glutamate-induced S-nitrosylation was blocked by 7NI. These results suggest that inactivation of CaMKII mediated by S-nitrosylation at Cys6 may contribute to NO-induced neurotoxicity in the brain. [ABSTRACT FROM AUTHOR]

Tissue plasminogen activator (tPA) is a serine protease that catalyzes the conversion of plasminogen (plg) to plasmin, which in turn functions to degrade extracellular matrix proteins in the central nervous system. The tPA-plasmin system plays a role in synaptic plasticity and remodeling. Here we show that this protease system participates in the rewarding effects of morphine by acutely regulating morphine-induced dopamine release in the nucleus accumbens (NAcc). A single morphine treatment induced tPA mRNA and protein expression in a naloxone-sensitive manner, which was associated with an increase in the enzyme activity in the NAcc. The acute effect of morphine in inducing tPA expression was diminished after repeated administration. Morphine-induced conditioned place preference and hyperlocomotion were significantly reduced in tPA-/- and plg-/- mice, being accompanied by a loss of morphine-induced dopamine release in the NAcc The defect of morphine-induced dopamine release and hyperlocomotion in tPA-/- mice was reversed by micro injections of either exogenous tPA or plasmin into the NAcc. Our findings demonstrate a previously undescribed function of the tPA-plasmin system in regulating dopamine release, which is involved in the rewarding effects of morphine. [ABSTRACT FROM AUTHOR]