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The information-theoretic waveform for target classification based on spectral variance has been studied, and its advantages have been shown. The waveform design algorithm has a low computational load and can be applied to real-time applications. In looking for more sophisticated methods while keeping the advantages of an information-based approach, a new design algorithm by deriving a strict lower bound of the information measure is developed. The proposed design algorithm requires a small amount of calculation and shows better classification performance in terms of percentage of correct detection. The proposed method is compared with other methods, and the improved performance is shown in numerically generated graphs.

The paper proposes a method for parallelizing migrations to reduce the time required for virtual machine (VM) relocation. During VM relocation, VMs need to wait for their migrations in a chain due to the limited resource of the physical machines (PMs), lengthening the VM relocation time. The essence of the proposed scheme is to break off the VMs' chained migration wait and enables VM migrations to be performed in parallel. The VMs' chained migration wait can be broken off by allowing some VMs, called breakup VMs, to migrate without waiting. A breakup VM is the first VM in a chain of migrations with which the total migration time is reduced when the chain is split at that VM and performed in parallel. Breakup VMs are selected recursively from all the split migration chains until the total migration time can no longer be reduced. After all breakup VMs are selected, VMs' chained migrations are parallelized by temporarily migrating the breakup VMs to spare PMs regardless of their initial migration order. The performance of the proposed scheme is evaluated through simulation. The VM relocation time is affected by the number of VMs and PM utilization in a cluster. Therefore, we simulate relocations of 100---800 VMs in clusters with different average CPU and memory utilizations of PMs. The simulation results show that the proposed scheme reduces VM relocation time by 21.9---62.0 % while using 1.6---5.8 % spare PMs to parallelize the chained migrations.

Despite the extensive use of propofol in general anesthetic procedures, the effects of propofol on glial cell were not completely understood. In lipopolysaccharide (LPS)-stimulated rat primary astrocytes and BV2 microglial cell lines, co-treatment of propofol synergistically induced inflammatory activation as evidenced by the increased production of NO, ROS and expression of iNOS, MMP-9 and several cytokines. Propofol augmented the activation of JNK and p38 MAPKs induced by LPS and the synergistic activation of glial cells by propofol was prevented by pretreatment of JNK and p38 inhibitors. When we treated BV2 cell culture supernatants treated with LPS plus propofol on cultured rat primary neuron, it induced a significant neuronal cell death. The results suggest that the repeated use of propofol in immunologically challenged situation may induce glial activation in brain.

Cytoplasmic polyadenylation binding protein 1 (CPEB1) is a RNA binding protein, which regulates translation of target mRNAs by regulating polyadenylation status. CPEB1 plays important roles in the regulation of germline cell development by modulating cell cycle progression through the polyadenylation of target mRNAs such as cyclin B1. Similar mechanism is reported in proliferating astrocytes by us, although CPEB1 is involved in the transport of target mRNAs as well as local translation at dendritic spines. In this study, we found the expression of CPEB1 in cultured rat primary neural progenitor cells (NPCs). EGF stimulation of cultured NPCs induced rapid phosphorylation of CPEB1, a hallmark of CPEB1-dependent translational control along with cyclin B1 polyadenylation and translation. EGF-induced activation of ERK1/2 and Aurora A kinase was responsible for CPEB1 phosphorylation. Pharmacological inhibition studies suggested that ERK1/2 is involved in the activation of Aurora A kinase and regulation of CPEB1 phosphorylation in cultured NPCs. Long-term incubation in EGF resulted in the down-regulation of CPEB1 expression, which further increased expression of cyclin B1 and cell cycle progression. When we down-regulated the expression of CPEB1 in NPCs by siRNA transfection, the proliferation of NPCs was increased. Increased NPCs proliferation by down-regulation of CPEB1 resulted in eventual up-regulation of neuronal differentiation with increase in both pre- and post-synaptic proteins. The results from the present study may suggest the importance of translational control in the regulation of neuronal development, an emerging concept in many neurodevelopmental and psychiatric disorders such as autism spectrum disorder.

Liver regeneration after liver damage caused by toxins and pathogens is critical for liver homeostasis. Retardation of liver proliferation was reported in hepatitis B virus (HBV) X protein (HBx)-transgenic mice. However, the underlying mechanism of the HBx-mediated disturbance of liver regeneration is unknown. We investigated the molecular mechanism of the inhibition of liver regeneration using liver cell lines and a mouse model. The mouse model of acute HBV infection was established by hydrodynamic injection of viral DNA. Liver regeneration after partial hepatectomy was significantly inhibited in the HBV DNA-treated mice. Mechanism studies have revealed that the expression of urokinase-type plasminogen activator (uPA), which regulates the activation of hepatocyte growth factor (HGF), was significantly decreased in the liver tissues of HBV or HBx-expressing mice. The down-regulation of uPA was further confirmed using liver cell lines transiently or stably transfected with HBx and the HBV genome. HBx suppressed uPA expression through the epigenetic regulation of the uPA promoter in mouse liver tissues and human liver cell lines. Expression of HBx strongly induced hypermethylation of the uPA promoter by recruiting DNA methyltransferase (DNMT) 3A2. Conclusion: Taken together, these results suggest that infection of HBV impairs liver regeneration through the epigenetic dysregulation of liver regeneration signals by HBx. (Hepatology 2013;58:762–776)

Although the role of α-synuclein aggregation on Parkinson's disease is relatively well known, the physiological role and the regulatory mechanism governing the expression of α-synuclein are unclear yet. We recently reported that α-synuclein is expressed and secreted from cultured astrocytes. In this study, we investigated the effect of valproic acid (VPA), which has been suggested to provide neuroprotection by increasing α-synuclein in neuron, on α-synuclein expression in rat primary astrocytes. VPA concentrationdependently increased the protein expression level of α-synuclein in cultured rat primary astrocytes with concomitant increase in mRNA expression level. Likewise, the level of secreted α-synuclein was also increased by VPA. VPA increased the phosphorylation of Erk1/2 and JNK and pretreatment of a JNK inhibitor SP600125 prevented the VPA-induced increase in α-synuclein. Whether the increased α-synuclein in astrocytes is involved in the reported neuroprotective effects of VPA awaits further investigation.

Plasminogen activator inhibitor-1 (PAI-1) is a member of serine protease inhibitor family, which regulates the activity of tissue plasminogen activator (tPA). In CNS, tPA/PAI-1 activity is involved in the regulation of a variety of cellular processes such as neuronal development, synaptic plasticity and cell survival. To gain a more insights into the regulatory mechanism modulating tPA/PAI-1 activity in brain, we investigated the effects of proteasome inhibitors on tPA/PAI-1 expression and activity in rat primary astrocytes, the major cell type expressing both tPA and PAI-1. We found that submicromolar concentration of MG132, a cell permeable peptide-aldehyde inhibitor of ubiquitin proteasome pathway selectively upregulates PAI-1 expression. Upregulation of PAI-1 mRNA as well as increased PAI-1 promoter reporter activity suggested that MG132 transcriptionally increased PAI-1 expression. The induction of PAI-1 downregulated tPA activity in rat primary astrocytes. Another proteasome inhibitor lactacystin similarly increased the expression of PAI-1 in rat primary astrocytes. MG132 activated MAPK pathways as well as PI3K/Akt pathways. Inhibitors of these signaling pathways reduced MG132-mediated upregulation of PAI-1 in varying degrees and most prominent effects were observed with SB203580, a p38 MAPK pathway inhibitor. The regulation of tPA/PAI-1 activity by proteasome inhibitor in rat primary astrocytes may underlie the observed CNS effects of MG132 such as neuroprotection.

J. Neurochem. (2012) 123, 226–238. Abstract Fragile X syndrome (FXS), the most common single genetic cause of mental retardation and autistic spectrum disease, occurs when FMR1 gene is mutated. FMR1 encodes fragile X mental retardation protein (FMRP) which regulates translation of mRNAs playing important roles in the development of neurons as well as formation and maintenance of synapses. To examine whether FMRP regulates cell viability, we induced apoptosis in rat primary cortical neurons with glutamate in vitro and with middle cerebral artery occlusion (MCAO) in striatal neurons in vivo. Both conditions elicited a rapid, but transient FMRP expression in neurons. This up-regulated FMRP expression was abolished by pre-treatment with PI3K and Protein Kinase B (Akt) inhibitors: LY294002, Akt inhibitor IV, and VIII. Reduced FMRP expression in vitro or in vivo using small hairpin Fmr1 virus exacerbated cell death by glutamate or MCAO, presumably via hypophosphorylation of Akt and reduced expression of B-cell lymphoma-extra large (Bcl-xL). However, over-expression of FMRP using enhanced green fluorescent protein (eGFP)-FMRP constructs alleviated cell death, increased Akt activity, and enhanced Bcl-xL production. The pro-survival role of Akt-dependent up-regulation of FMRP in glutamate-stimulated cultured neuron as well as in ischemic brain may have a clinical importance in FXS as well as in neurodegenerative disorders and traumatic brain injury.

Oroxylin A is a flavone isolated from a medicinal herb reported to be effective in reducing the inflammatory and oxidative stresses. It also modulates the production of brain derived neurotrophic factor (BDNF) in cortical neurons by the transactivation of cAMP response element-binding protein (CREB). As a neurotrophin, BDNF plays roles in neuronal development, differentiation, synaptogenesis, and neural protection from the harmful stimuli. Adenosine A2A receptor colocalized with BDNF in brain and the functional interaction between A2A receptor stimulation and BDNF action has been suggested. In this study, we investigated the possibility that oroxylin A modulates BDNF production in cortical neuron through the regulation of A2A receptor system. As ex-pected, CGS21680 (A2A receptor agonist) induced BDNF expression and release, however, an antagonist, ZM241385, prevented oroxylin A-induced increase in BDNF production. Oroxylin A activated the PI3K-Akt-GSK-3β signaling pathway, which is inhibited by ZM241385 and the blockade of the signaling pathway abolished the increase in BDNF production. The physiological roles of oroxylin A-induced BDNF production were demonstrated by the increased neurite extension as well as synapse formation from neurons. Overall, oroxylin A might regulate BDNF production in cortical neuron through A2A receptor stimulation, which promotes cellular survival, synapse formation and neurite extension.

This letter introduces an energy sensitive bandpass filter (ESBPF) to protect Ku-band low noise amplifiers (LNAs) from high power electromagnetic (HPEM) threats. The ESBPF circuit has anti-parallel Schottky Barrier diodes mounted on a planar bandpass filter (BPF) circuit. The ESBPF operates as a BPF at a power level below the maximum permissible power level (MPPL) of the LNAs. However, the circuit works like a variable attenuator at the power level equal to or higher than the MPPL of the LNAs. To increase attenuation and selectivity functions, a 63 $^{\circ}$ line section between two planar filter circuits loaded transversely along WR-75 waveguide has been inserted to cascade. The development of the circuit model has been started with lumped elements under the condition of a 0 dBm MPPL of LNA. Then, the model has been simulated, optimized with HFSS, and fabricated. Measurement results show that the ESBPF has insertion loss less than 1.27 dB at the power level lower than $-$ 2 dBm for the frequency range from 11.8 to 12.3 GHz. At the power level higher than $-$ 2 dBm, the circuit provides different levels of attenuation depending on the input power within the identical frequency band; 31 dB insertion loss, which provides isolation characteristic, has been measured at the power level of 30 dBm.

Odontogenic keratocyst is one of the most aggressive odontogenic cyst due to its relatively high recurrence rate, fast growth, and its tendency to invade adjacent tissue. The aim of the present study is to analyze retrospectively the clinico-pathological characteristics of the 220 KCOTs cases in 181 and find the relationship among the various factors. And the following data were obtained: patient age ranged from 8 to 77 years with an average of 33.06 years. There were 111 male and 70 female patients. The mandibular angle and ascending ramus was the most frequent site of KCOTs. Swelling, the most presenting sign, was noted in 97 cases, followed by the pain, 85 cases, and 78 cases showed no presenting signs and symptoms. On radiologic findings, the primary KCOTs appeared as a unilocular in 59 cases or multilocular in 39 cases. The KCOTs can achieve the size of which may vary from 1.0 cm to 13.0 cm, and the mean size was 4.07 cm. The KCOTs were associated with the displacement of impacted teeth, the mandibular third molar; 47 cases, was the most frequent impacted teeth. Four patients (2.21%) were confirmed to be associated with nevoid basal cell carcinoma syndrome. Enucleation or curettage was the most common employed treatment modality (164 cases), followed by marsupialization or decompression (17 cases), and lateral decortication (13 cases). The recurrence rate was 10.78%. Most of the recurrences (21 cases) were found during the first 5 years after the first treatment for the KCOTs.

Prenatal exposure to valproic acid (VPA) induces neural tube defects and impairment in social behaviors related to autistic spectrum disorder in newborns, which make it a useful animal model of autism. In this study, we compared the effects of different time window of prenatal valproic acid exposure for inducing the altered social behaviors relevant to autism from embryonic day 7 to embryonic day 15 in Sprague–Dawley rats to determine the critical periods for the impairment. Compared to E7, E9.5 and E15 exposure, VPA exposure at E12 showed most significant changes in behaviors over control animals with reduced sociability and social preference. E9.5 exposure to valproic acid showed strong reproductive toxicity including decrease in the number of live birth. In general, exposure at E15 showed only marginal effects on reproduction and social behaviors. Finally, VPA-exposed rats at E12 were more sensitive to electric shock than VPA-exposed rats at any other periods. These results suggested that E12 is the critical period in rats when valproate exposure has prominent effects for inducing the altered social behavior similar to human autistic behavior.

This letter presents the field uniformity characteristics of a triangular prism reverberation chamber. A reverberation chamber that generally uses a stirrer to create a uniform electric field inside is an alternative to the semi-anechoic chamber for an electromagnetic compatibility test. To overcome the size and maintenance problems of a stirrer, we propose to replace it with a Quadratic Residue Diffuser which is commonly used in acoustics. To confirm that the diffuser is a valid alternative to the stirrer, a diffuser and an equilateral triangular prism reverberation chamber are designed and fabricated for 2.3-3.0GHz operation. To investigate the field uniformity characteristics by varying the location of the transmitting antenna, both simulation and measurement in the triangular prism reverberation chamber were also done at its two positions, respectively. A commercial program XFDTD 6.2, engaging the finite difference time domain (FDTD), is used for simulation and a cumulative probability distribution, which the IEC 61000-4-21 recommends, is used to evaluate the field uniformity. Both simulation and measurement results show that the field uniformity in the chamber satisfies the international standard requirement of ±6dB tolerance and ±3dB standard deviation, which means that a diffuser can be substituted for a stirrer.

Actual evapotranspiration (AET) in the Suyeong-gu was estimated and correlations between AET and meteorological factors were analyzed. The study area was Suyeong-gu lay at the east longitude 05' 40" ~ 129 08' 08" and north latitude 07' 59" ~ 11' 01". The Kumryun mountain, the Bae mountain, the Suyeong river and the Suyeong bay are located on west, north, northeaster and south side in the study area, respectively. AET was estimated using precipitation (P), potential evapotranspiration (PET) and plant-available water coefficient. Meteorological factors to estimate PET were air temperature, dewpoint temperature, atmospheric pressure, duration of sunshine and mean wind speed (MWS). PET and AET were estimated by a method of Allen et al. (1998) and Zhang et al. (2001), respectively. PET was the highest value (564.45 mm/yr) in 2002 year, while it was the lowest value (449.95 mm/yr) in 2003 year. AET was estimated highest value (554.14 mm/yr) in 2002 year and lowest value (427.91 mm/yr) in 2003 year. Variations of PET and AET were similar. The linear regression function of AET as PET using monthly data was AET=0.87PET+3.52 and coefficient of determination was high, 0.75. In order to analyze relationship between the evapotranspiration and meteorological factors, correlation analysis using monthly data were accomplished. Correlation coefficient of AET-PET was 0.96 high, but they of AET-P and PET-P were very low. Correlation coefficients of AET-MWS and PET-MWS were 0.67 and 0.73, respectively. Thus, correlation between evapotranspiration and MWS was the highest among meteorological factors in Suyong-gu. This means that meteorological factor to powerfully effect for the variation of evapotranspiration was MWS. The linear regression function of AET as MWS was AET=84.73MWS+223.05 and coefficient of determination was 0.54. The linear regression function of PET as MWS was PET=83.83MWS+203.62 and coefficient of determination was 0.45.

This paper presents a method of expanding the operating frequency band of a Reverberating TEM Cell (RTC) for electromagnetic compatibility (EMC) testing. To expand the operating frequency band of an RTC, this paper places a wire septum inside the cell instead of a solid septum. The maximum usable frequency (MUF) for TEM cell operation and the lowest usable frequency (LUF) for reverberating chamber operation with the wire septum are studied and compared with a conventional solid septum. The E field strengths inside the RTC are measured and evaluated. The measurement results show that the RTC with the wire septum have similar MUF to the RTC with a solid septum at TEM mode, but have much lower LUF at a reverberating mode, which proves that the operating frequency band of the RTC can be expanded by using the wire septum.

Protease-activated receptors (PARs) play important roles in the regulation of brain function such as neuroinflammation by transmitting the signal from proteolytic enzymes such as thrombin and trypsin. We and others have reported that a member of the family, PAR-2 is activated by trypsin, whose involvement in the neurophysiological process is increasingly evident, and is involved in the neuroinflammatory processes including morphological changes of astrocytes. In this study, we investigated the role of PAR-2 in the production of nitric oxide (NO) in rat primary astrocytes. Treatment of PAR-2 agonist trypsin increased NO production in a dose-dependent manner, which was mediated by the induction of inducible nitric-oxide synthase. The trypsin-mediated production of NO was mimicked by PAR-2 agonist peptide and reduced by either pharmacological PAR-2 antagonist peptide or by siRNA-mediated inhibition of PAR-2 expression, which suggests the critical role of PAR-2 in this process. NO production by PAR-2 was mimicked by PMA, a PKC activator, and was attenuated by Go6976, a protein kinase C (PKC) inhibitor. PAR-2 stimulation activated three subtypes of mitogen-activated protein kinases (MAPKs): extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 MAPK. NO production by PAR-2 was blocked by inhibition of ERK, p38, and JNK pathways. PAR-2 stimulation also activated nuclear factor-kappaB (NF-kappaB) DNA binding and transcriptional activity as well as IkappaBalpha phosphorylation. Inhibitors of NF-kappaB pathway inhibited PAR-2-mediated NO production. In addition, inhibitors of MAPK pathways prevented transcriptional activation of NF-kappaB reporter constructs. These results suggest that PAR-2 activation-mediated NO production in astrocytes is transduced by the activation of MAPKs followed by NF-kappaB pathways.

In this paper, we propose an automatic extraction model for unknown translations and implement an unknown translation extraction system using the proposed model. The proposed model as a phrase-alignment model is incorporated with three models: a phrase-boundary model, a language model, and a translation model. Using the proposed model we implement the system for extracting unknown translations, which consists of three parts: construction of parallel corpora, alignment of Korean and English words, extraction of unknown translations. To evaluate the performance of the proposed system we have established the reference corpus for extracting unknown translation, which comprises of 2,220 parallel sentences including about 1,500 unknown translations. Through several experiments, we have observed that the proposed model is very useful for extracting unknown translations. In the future, researches on objective evaluation and establishment of parallel corpora with good quality should be performed and studies on improving the performance of unknown translation extraction should be kept up.

We introduce a new information theoretic power allocation scheme applicable to distributed multiple-hypothesis detection systems communicating over slow fading channels. In earlier work, it was demonstrated that performance could be improved by adjusting transmit power to maximize the J-divergence measure of a binary detection system and the J-divergence method is extended for a distributed multiple-hypothesis detection system by defining pairwise sums of the J-divergences. However, the pairwise sum measure does not provide a tight bound. Basically, the more hypotheses we adopt, the less efficient the optimization is. Thus, we derive a more efficient classification-oriented information measure for power optimization of distributed multiple-hypothesis system by introducing a virtual decider variable. The virtual decider variable is directly related with classification task. Various numerical results are also shown to compare the performances.

Recently, Montemurro et al. showed that there would be a long-range correlation in Shakespeare’s 36 plays written in English. However it is not clear that similar result can be found in other language which has very different grammar structure from English. So we examine whether there exists a long-range correlation in Shakespeare’s plays translated into Korean which would have very different syntactic rules from English. Additionally, we perform the same experiment for 12 novels written by four Korean popular writers in order to see the difference due to the styles of the writers. As a result, we find Hurst’s exponents of 0.632 ± 0.03 in Shakespeare’s plays translated in Korean. Also we find the similar values of Hurst’s exponent even to the novels of the Korean popular writers. It implies that regardless of languages, there exists a long-range correlation in literary corpora.

Although ubiquitously expressed, the transcriptional factor CP2 also exhibits some tissue- or stage-specific activation toward certain genes such as globin in red blood cells and interleukin-4 in T helper cells. Because this specificity may be achieved by interaction with other proteins, we screened a peptide display library and identified four consensus motifs in numerous CP2-binding peptides: HXPR, PHL, ASR and PXHXH. Protein-database searching revealed that RE-1 silencing factor (REST), Yin-Yang1 (YY1) and five other proteins have one or two of these CP2-binding motifs. Glutathione S-transferase pull-down and coimmunoprecipitation assays showed that two HXPR motif-containing proteins REST and YY1 indeed were able to bind CP2. Importantly, this binding to CP2 was almost abolished when a double amino acid substitution was made on the HXPR sequence of REST and YY1 proteins. The suppressing effect of YY1 on CP2's transcriptional activity was lost by this point mutation on the HXPR sequence of YY1 and reduced by an HXPR-containing peptide, further supporting the interaction between CP2 and YY1 via the HXPR sequence. Mapping the sites on CP2 for interaction with the four distinct CP2-binding motifs revealed at least three different regions on CP2. This suggests that CP2 recognizes several distinct binding motifs by virtue of employing different regions, thus being able to interact with and regulate many cellular partners.

Speech features of the conventional speaker identification system, are usually obtained by linear methods in spectral space. However, these methods have the drawback that speakers with similar voices cannot be distinguished, because the characteristics of their voices are also similar in spectral space. To overcome the difficulty in linear methods, we propose to use the correlation exponent in the nonlinear space as a new feature vector for speaker identification among persons with similar voices. We show that our proposed method surprisingly reduces the error rate of speaker identification system to speakers with similar voices.

Eight 2-alkylaminosubstituted 5,8-dimethoxy-4-methylquinolines and nine 2-alkylaminosubstituted or 2,6-disubstituted 4-methylquinoline-5,8-diones were synthesized and evaluated in vitro cytotoxicity against four human cancer cell lines (HOP62, SK-OV-3, HCT15 and SF295).

The immediate effect of a 15-min heat shock was examined on the level and the activity of Akt. Following heat shock, the Akt level decreased by 15-70% in a temperature-dependent and phosphorylation status-independent manner. This decrease of Akt level was not prevented by caspase inhibitors. At 48 degrees C, the extent of the breakdown was so immense that the total phosphorylation/activity level of Akt was not increased over the control level, implying that the total cellular activity of Akt governed by the level and the molar activity does not necessarily undergo the ensuing change.

Five 1-azaanthraquinone-3-carboxamides were synthesized and evaluated in vitro cytotoxicity against four human cancer cell lines. The most active compound, 7b, exhibited cytotoxic activity comparable to doxorubicin.

A series of benzo[g]quinoxaline-5,10-dione derivatives carrying a 7-dialkylaminomethyl substituent was synthesized and their in vitro cytotoxic activities were evaluated against four human cancer cell lines (HCT-15, SK-OV-3, MD-MB-468 and T-47D). The most active compound 9d showed cytotoxic activity comparable to that of doxorubicin against HCT-15 cancer cell line.

The serine/threonine protein kinase encoded by the Akt proto-oncogene is catalytically inactive in serum-starved primary and immortalized fibroblasts. Here we show that Akt and the Akt-related kinase AKT2 are activated by PDGF. The activation was rapid and specific, and it was abrogated by mutations in the Akt Pleckstrin homology (PH) domain. The Akt activation was also shown to depend on PDGFRβ tyrosines Y740 and Y751, which bind phosphatidylinositol 3-kinase (PI 3-kinase) upon phosphorylation. Moreover, Akt activation was blocked by the PI 3-kinase-specific inhibitor wortmanin and the dominant inhibitory N17Ras. Conversely, Akt activity was induced following the addition of phosphatidylinositol-3-phosphate to Akt immunoprecipitates from serum-starved cells in vitro. These results identify Akt as a novel target of PI 3-kinase and suggest that the Akt PH domain may be a mediator of PI 3-kinase signaling.

The cytoplasmic serine-threonine protein kinase coded for by the c-akt proto-oncogene features a protein kinase C-like catalytic domain and a unique NH2-terminal domain (AH domain). The AH domain is a member of a domain superfamily whose prototype was observed in pleckstrin (pleckstrin homology, or PH, domain). In this communication, we present evidence that the AH/PH domain is a domain of protein-protein interaction which mediates the formation of Akt protein complexes. The interaction between c-akt AH/PH domains is highly specific, as determined by the failure of this domain to bind AKT2. The AH/PH domain-mediated interactions depend on the integrity of the entire domain. Akt molecules with deletions of the NH2-terminal portion (amino acids 11 to 60) and AH/PH constructs with deletions of the C-terminal portion of this domain (amino acids 107 to 147) fail to interact with c-akt. To determine the significance of these findings, we carried out in vitro kinase assays using Akt immunoprecipitates from serum-starved and serum-starved, platelet-derived growth factor-stimulated NIH 3T3 cells. Addition of maltose-binding protein-AH/PH fusion recombinant protein, which is expected to bind Akt, to the immunoprecipitates from serum-starved cells induced the activation of the Akt kinase.

J. Neurochem. (2012) 123, 226-238.Fragile X syndrome (FXS), the most common single genetic cause of mental retardation and autistic spectrum disease, occurs when FMR1 gene is mutated. FMR1 encodes fragile X mental retardation protein (FMRP) which regulates translation of mRNAs playing important roles in the development of neurons as well as formation and maintenance of synapses. To examine whether FMRP regulates cell viability, we induced apoptosis in rat primary cortical neurons with glutamate in vitro and with middle cerebral artery occlusion (MCAO) in striatal neurons in vivo. Both conditions elicited a rapid, but transient FMRP expression in neurons. This up-regulated FMRP expression was abolished by pre-treatment with PI3K and Protein Kinase B (Akt) inhibitors: LY294002, Akt inhibitor IV, and VIII. Reduced FMRP expression in vitro or in vivo using small hairpin Fmr1 virus exacerbated cell death by glutamate or MCAO, presumably via hypophosphorylation of Akt and reduced expression of B-cell lymphoma-extra large (Bcl-xL). However, over-expression of FMRP using enhanced green fluorescent protein (eGFP)-FMRP constructs alleviated cell death, increased Akt activity, and enhanced Bcl-xL production. The pro-survival role of Akt-dependent up-regulation of FMRP in glutamate-stimulated cultured neuron as well as in ischemic brain may have a clinical importance in FXS as well as in neurodegenerative disorders and traumatic brain injury.

Protein phosphatase 2A (PP2A) is composed of structural (A), catalytic (C), and regulatory subunits (B). Immunological analyses identified B alpha/PR55 alpha as the major regulatory subunit of brain PP2A while a unique B' subunit was associated with the cardiac enzyme. Recombinant PP2A heterotrimers were purified from insect cells infected with baculoviruses expressing A and C, in combination with viruses expressing B alpha/PR55 alpha, B beta/PR55 beta, or SV40 small tumor antigen (st). Phosphatase activities of rAC-B alpha and rAC-B beta were similar to those for brain AC-B alpha, while rAC-st was 50-80% less active. Heparin had no effect on rAC-st myosin light chain phosphatase activity, while the B subunit-containing forms were stimulated 2-3-fold. Protamine caused a 3-4-fold increase in AC-B alpha and rAC-st activities and a marked activation of rAC-B beta (6-fold) and AC-B' (10.5-fold). When histone H1 was used as substrate, all of the heterotrimers were stimulated approximately 4-fold by heparin. The activity of AC-B' and rAC-B beta were increased 2-fold by Mn2+, while a 6-fold stimulation was observed with rAC-st. Chemical cross-linking of AC-B alpha and AC-B beta generated 200-kDa complexes, while AC-st was present as a 150-kDa complex. These results demonstrate that different regulatory proteins affect enzyme activity and the response to agents that modify PP2A activity in vitro. Different PP2A heterotrimers are likely to have distinct functions in vivo, and changes in subunit composition will have an important impact on signal transduction pathways.

As a member of neurotrophin family, brain derived neurotrophic factor (BDNF) plays critical roles in neuronal development, differentiation, synaptogenesis, and neural protection from the harmful stimuli. There have been reported that adenosine A2(A) receptor subtype is widely distributed in the brain regions, such as hippocampus, striatum, and cortex. Adenosine A2(A) receptor is colocalized with BDNF in brain regions and the functional interaction between A2(A) receptor stimulation and BDNF action has been suggested. In this study, we investigated the possibility that the activation of A2(A) receptor modulates BDNF production in rat primary cortical neuron. CGS21680, an adenosine A2(A) receptor agonist, induced BDNF expression and release. An antagonist against A2(A) receptor, ZM241385, prevented CGS21680-induced increase in BDNF production. A2(A) receptor stimulation induced the activation of Akt-GSK-3β signaling pathway and the blockade of the signaling pathway with specific inhibitors abolished the increase in BDNF production, possibly via modulation of ERK1/2-CREB pathway. The physiological roles of A2(A) receptor-induced BDNF production was demonstrated by the protection of neurons from the excitotoxicity and increased neurite extension as well as synapse formation from immature and mature neurons. Taken together, activation of A2(A) receptor regulates BDNF production in rat cortical neuron, which provides neuro-protective action.

Background Fragile X syndrome (FXS), the most commonly inherited mental retardation and single gene cause of autistic spectrum disorder, occurs when the Fmr1 gene is mutated. The product of Fmr1, fragile X linked mental retardation protein (FMRP) is widely expressed in HeLa cells, however the roles of FMRP within HeLa cells were not elucidated, yet. Interacting with a diverse range of mRNAs related to cellular survival regulatory signals, understanding the functions of FMRP in cellular context would provide better insights into the role of this interesting protein in FXS. Using HeLa cells treated with etoposide as a model, we tried to determine whether FMRP could play a role in cell survival. Methods Apoptotic cell death was induced by etoposide treatment on Hela cells. After we transiently modulated FMRP expression (silencing or enhancing) by using molecular biotechnological methods such as small hairpin RNA virus-induced knock down and overexpression using transfection with FMRP expression vectors, cellular viability was measured using propidium iodide staining, TUNEL staining, and FACS analysis along with the level of activation of PI3K-Akt pathway by Western blot. Expression level of FMRP and apoptotic regulator BcL-xL was analyzed by Western blot, RT-PCR and immunocytochemistry. Results An increased FMRP expression was measured in etoposide-treated HeLa cells, which was induced by PI3K-Akt activation. Without FMRP expression, cellular defence mechanism via PI3K-Akt-Bcl-xL was weakened and resulted in an augmented cell death by etoposide. In addition, FMRP over-expression lead to the activation of PI3K-Akt signalling pathway as well as increased FMRP and BcL-xL expression, which culminates with the increased cell survival in etoposide-treated HeLa cells. Conclusions Taken together, these results suggest that FMRP expression is an essential part of cellular survival mechanisms through the modulation of PI3K, Akt, and Bcl-xL signal pathways.

A hetero Diels-Alder reaction of quinoline-5,8-dione with 1-(N,N-dimethylamino)-3-methyl-1-aza-1,3-butadiene proceeded to give 3-methyl-1,8-diazaanthraquinone (100% regioselectivity) in the presence of Lewis-acid catalyst (ZnCl2 or ZnBr2). Subsequent functionalizations of the benzylic methyl group resulted in the 1,8-diazaanthraquinone analogues as potential antitumor agents. The most active compound, 8, exhibited in vitro cytotoxic activity comparable to that of doxorubicin.