Cdk5 links with DNA damage response and cancer.

As an atypical member of cyclin dependent kinase family, Cyclin dependent kinase 5 (Cdk5) is considered as a neuron-specific kinase in the past decade due to the abundant existence of its activator p35 in post-mitotic neurons. Recent studies show that Cdk5 participates in a series of biological and pathological processes in non-neuronal cells, and is generally dysregulated in various cancer cells. The inhibition or knockdown of Cdk5 has been proven to play an anti-cancer role through various mechanisms, and can synergize the killing effect of chemotherapeutics. DNA damage response (DDR) is a series of regulatory events including DNA damage, cell-cycle arrest, regulation of DNA replication, and repair or bypass of DNA damage to ensure the maintenance of genomic stability and cell viability. Here we describe the regulatory mechanisms of Cdk5, its controversial roles in apoptosis and focus on its links to DDR and cancer.

Calcium signal-initiated early activation of NF-kappaB in neurons is a neuroprotective event in response to kainic acid-induced excitotoxicity.

We demonstrate that activation of nuclear factor kappaB (NF-kappaB) in neurons is neuroprotective in response to kainic acid (KA)-induced excitotoxicity. Combination of Western blotting, immunocytochemistry, and electrophoresis mobility shift assay showed that KA exposure induced a fast but transient nuclear translocation of the NF-kappaB p65 subunit and increased DNA-binding activity of NF-kappaB in primary cultured cortical neurons. The transient NF-kappaB activity was associated with upregulation of antiapoptotic Bcl-xL and XIAP gene products revealed by real-time PCR. Knockdown of p65 decreased neuronal viability and antiapoptotic gene expression. In addition, we showed that KA-stimulated DNA-binding activity of NF-kappaB was associated with reactive oxygen species and calcium signals, using AMPA/KA receptor antagonist, calcium chelator, and antioxidant. These results suggest that the fast and transient activation of NF-kappaB initiated by calcium signals is one of the important proximal events in response to KA-induced excitotoxicity, which has neuroprotective effect against KA-induced apoptosis.

Continuous bioelectricity production and sustainable wastewater treatment in a microbial fuel cell constructed with non-catalyzed granular graphite electrodes and permeable membrane.

Oxygen has been so far addressed as the most preferable terminal electron acceptor in the cathodes of microbial fuel cells (MFCs). However, to reduce the oxygen reduction overpotential at the cathode surface, eco-unfriendly and costly catalysts have been commonly employed. Here, we pursued the possibility of using a high surface area electrode to reduce the cathodic reaction overpotential rather than the utilization of catalyzed materials. A dual chambered MFC reactor was designed with the use of graphite-granule electrodes and a permeable membrane. The performance of the reactor in terms of electricity generation and organic removal rate was examined under a continuous-feed manner. Results showed that the maximum volumetric power of 4.4+/-0.2 W/m(3) net anodic compartment (NAC) was obtained at a current density of 11+/-0.5 A/m(3) NAC. The power output was improved by increasing the electrolyte ionic strength. An acceptable effluent quality was attained when the organic loading rate (OLR) of 2 kgCOD/m(3) NAC d was applied. The organic removal rate seemed to be less affected by shock loading. Our system can be suggested as a promising approach to make MFC-based technology economically viable for wastewater treatment applications. This study shows that current generation can be remarkably improved in comparison with several other studies using a low-surface-area plain graphite electrode.

Kainate exposure suppresses activation of GluR2 subunit promoter in primary cultured cerebral cortical neurons through induction of RE1-silencing transcription factor.

The AMPA receptor subunit GluR2 is downregulated in neurons following a wide range of neurological insults. Here we report that suppression of GluR2 gene promoter activity is associated with kainate (KA)-induced downregulation of GluR2 subunit levels in primary cultured cortical neurons. RT-PCR and Northern blotting showed a significant decrease in GluR2 mRNA in cultured neurons after KA exposure. Transfection of cultured neurons with an expression vector pGL3-GluR2(-298/+283), where the reporter gene firefly luciferase was driven by the GluR2 promoter, revealed that KA exposure suppressed the transcriptional activation of the GluR2 promoter. Furthermore, the expression of the RE1-silencing transcription factor (REST) was increased in KA-exposed cortical neurons; enhanced binding of REST to RE1-like silencer element in the proximal promoter of the GluR2 subunit gene was evidenced by electrophoresis mobility shift assay. Chromatin immunoprecipitation showed that suppressed activity of the GluR2 promoter in cultured neurons after KA exposure was related to deacetylation of histone H4. These results indicate that REST as a crucial factor binds to RE1-like silencer element in the GluR2 promoter, suppressing transcription of the GluR2 subunit gene during KA exposure. Our data suggest that transcriptional suppression of the GluR2 subunit gene may contribute at least in part to downregulation of GluR2 subunit protein in neurons during KA exposure. Because our experiments showed a reduction of glutamate release in KA-exposed cortical neurons, REST may play a latent role in delayed neuronal death or in seizure-induced tolerance.

Exposure of human lung cancer cells to 8-chloro-adenosine induces G2/M arrest and mitotic catastrophe.

8-Chloro-adenosine (8-Cl-Ado) is a potent chemotherapeutic agent whose cytotoxicity in a variety of tumor cell lines has been widely investigated. However, the molecular mechanisms are uncertain. In this study, we found that exposure of human lung cancer cell lines A549 (p53-wt) and H1299 (p53-depleted) to 8-Cl-Ado induced cell arrest in the G2/M phase, which was accompanied by accumulation of binucleated and polymorphonucleated cells resulting from aberrant mitosis and failed cytokinesis. Western blotting showed the loss of phosphorylated forms of Cdc2 and Cdc25C that allowed progression into mitosis. Furthermore, the increase in Ser10-phosphorylated histone H3-positive cells revealed by fluorescence-activated cell sorting suggested that the agent-targeted cells were able to exit the G2 phase and enter the M phase. Immunocytochemistry showed that microtubule and microfilament arrays were changed in exposed cells, indicating that the dynamic instability of microtubules and microfilaments was lost, which may correlate with mitotic dividing failure. Aberrant mitosis resulted in mitotic catastrophe followed by varying degrees of apoptosis, depending on the cell lines. Thus, 8-Cl-Ado appears to exert its cytotoxicity toward cells in culture by inducing mitotic catastrophe.

[Effects of the rice straw on Microcystis aeraginosa analyzed by different physiological parameters].

The growth and physiology of bloom-forming cyanobacterium Microcystis aeruginosa were determined by the flow cytometry when exposed to rice straw extract for 15 d. The cell growth, cell integrity, mitochondrial transmembrane potential, and esterase activity were used to evaluate the physiological response in Microcystis aeruginosa. Rice straw extract stored for 5 days significantly inhibited the growth of Microcystis aeruginosa in a concentration-dependent way; Most of the algae cells (> 98%) remained complete membranes in all the concentration treatments; Compared with the control cultures, the rice straw induced both negative and positive effects on the esterase activity for each test within 4 days, while the inhibition exceeded the stimulation effect. After a 7 d exposure, only the inhibition effect was found. Neither the inhibited nor stimulated effects was observed after algae exposure from 10 d to 15 d. Evident changes was found in the membrane potential during 7 d experiment, whereas inhibition effect became weaker after 10 d and 15 d exposure, in consistent with the result of esterase activity. These results confirmed that the rice straw extract might provide both dominant inhibition and relatively weak stimulation effects. After a long time exposure, inhibition effect became limited while stimulation effect disappeared. The action of rice straw may be algistatic (preventing algal growth) but not algicidal (killing algae).

Stabilization of mitochondrial function by tetramethylpyrazine protects against kainate-induced oxidative lesions in the rat hippocampus.

Mitochondria are critical regulators of cell death, a key feature of neurodegeneration. Reactive oxygen species (ROS) are crucial to Ca(2+)-mediated effects of glutamate receptor activation leading to neuronal degeneration. Tetramethylpyrazine (TMP) is a principal ingredient of Ligusticum wallichi Franchat (a Chinese herb), used for treatment of cardiovascular and cerebrovascular ischemic diseases. However, its protection against oxidative brain injury associated with excessive activation of glutamate receptors is unknown. In this study, we demonstrate TMP neuroprotection against kainate-induced excitotoxicity in vitro and in vivo. We found that TMP could partly alleviate kainate-induced status epilepticus in rats and prevented and rescued neuronal loss in the hippocampal CA3 but not the CA1 region. The partial prevention and rescue of neuronal loss by TMP were attributable to the preservation of the structural and functional integrity of mitochondria, evidenced by maintaining the mitochondrial membrane potential, ATP production, and complex I and III activities. Stabilization of mitochondrial function was linked to the observation that TMP could function as a reductant/antioxidant to quench ROS, block lipid peroxidation, and protect enzymatic antioxidants such as glutathione peroxidase and glutathione reductase. These results suggest that TMP may protect against oxidative brain injury by stabilization of mitochondrial function through quenching of ROS.

Dynamics of augmented soil system containing biphenyl with Dyellaginsengisoli LA-4.

One high efficient biphenyl-degrading strain Dyella ginsengisoli LA-4 was inoculated into biphenyl-contaminated soil for bioaugmentation in this study. The results showed that bioaugmentation could accelerate the startup period of the biphenyl bioremediation process compared with the non-augmented one. PCR-DGGE fingerprints demonstrated that both of the diversity and pattern of microbial community were affected by the addition of strain LA-4 and biphenyl. Biphenyl-utilizing populations gradually increased and become the dominant species. The introduced strain LA-4 could be persistent and co-exist well with the indigenous populations. However, both of the strain LA-4 and indigenous microorganisms in the bioaugmented system would be partially inhibited by Zn(2+) and Ni(2+). This study suggests that it is feasible and potentially useful to remediate biphenyl-contaminated soil using bioaugmentation with D. ginsengisoli LA-4.

Simultaneous organics removal and bio-electrochemical denitrification in microbial fuel cells.

Simultaneous organics removal and bio-electrochemical denitrification using a microbial fuel cell (MFC) reactor were investigated in this study. The electrons produced as a result of the microbial oxidation of glucose in the anodic chamber were transferred to the anode, which then flowed to the cathode in the cathodic chamber through a wire, where microorganisms used the transferred electrons to reduce the nitrate. The highest power output obtained on the MFCs was 1.7 mW/m(2) at a current density of 15 mA/m(2). The maximum volumetric nitrate removal rate was 0.084 mg NO(3)(-)-N cm(-2) (electrode surface area) day(-1). The coulombic efficiency was about 7%, which demonstrated that a substantial fraction of substrate was lost without current generation.