Apoptosis induction on human hepatoma cells Hep G2 of decabrominated diphenyl ether (PBDE-209)

Polybrominated diphenyl ethers (PBDEs) are an important class of halogenated organic brominated flame retardants. Because of their presence in abiotic and biotic environments widely and their structural similarity to polychlorinated biphenyls (PCBs), concern has been raised on their possible adverse health effects to humans. This study was designed to determine the anti-proliferative, apoptotic properties of decabrominated diphenyl ether (PBDE-209), using a human hepatoma Hep G2 line as a model system. Hep G2 cells were cultured in the presence of PBDE-209 at various concentrations (1.0-100.0 mu mol/L) for 72 h and the percentage of cell viability was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The results showed that PBDE-209 inhibited the cells viability in time and concentration-dependent characteristics at concentrations (10.0-100.0 mu mol/L). We found that anti-proliferative effect of PBDE-209 was associated with apoptosis on Hep G2 cells by determinations of morphological changes, cell cycle and apoptosis. Mechanism study showed that PBDE-209 could increase the generation of intracellular reactive oxygen species (ROS) concentration-dependently. Antioxidant N-acetylcyteine partially inhibited the increase of ROS. The mechanism for its hepatoma-inhibitory effects was the induction of cellular apoptosis through ROS generation. In addition, activity of lactate dehydrogenase (LDH) release increased when the cells incubated with PBDE-209 at various concentrations and times. These results suggested that PBDE-209 had the toxicity activity of anti-proliferation and induction of apoptosis in tumor cells in vitro. (c) 2007 Elsevier Ireland Ltd. All rights reserved.

Induction of oxidative stress and apoptosis by PFOS and PFOA in primary cultured hepatocytes of freshwater tilapia (Oreochromis niloticus)

Perfluorinated organic compounds (PFOCs) are emerging persistent organic pollutants (POPs) widely present in the environment, wildlife and human. We studied the cellular toxicology of perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) on oxidative stress and induction of apoptosis in primary cultured hepatocytes of freshwater tilapia (Oreochromis niloticus). Cultured hepatocytes were exposed to PFOS or PFOA (0, 1, 5, 15 and 30 mg L-1) for 24 h, and a dose-dependent decrease in cell viability was determined using trypan blue exclusion method. Significant induction of reactive oxygen species (ROS) accompanied by increases in activities of superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) were found, while activities of glutathione peroxidase (GPx) and glutathione-S-transferase (GST) were decreased. Glutathione (GSH) content was reduced following treatment of PFOA and PFOS. A dose-dependent increase in the lipid peroxidation (LPO) level (measured as maleic dialdehyde, MDA) was observed only in the PFOA exposure groups, whereas LPO remained unchanged in the PFOS exposure groups. Furthermore, a significant activation of caspase-3, -8, -9 activities was evident in both PFOS and PFOA exposure groups. Typical DNA fragmentation (DNA laddering) was further characterized by agarose gel electrophoresis. The overall results demonstrated that PFOS and PFOA are able to produce oxidative stress and induce apoptosis with involvement of caspases in primary cultured tilapia hepatocytes. (c) 2007 Elsevier B.V. All rights reserved.

Vitellogenin in rare minnow (Gobiocypris rarus): identification and induction by waterborne diethylstilbestrol

Rare minnow (Gobiocypris rarus) is a tiny Chinese carp that has a short life cycle and is easily cultured in the laboratory. In this study, juvenile rare minnows were exposed to waterborne diethylstilbestrol (DES) at 0.05, 0.5 and 5 mug/l in laboratory aquaria. After exposure for 4, 8, 13 and 21 days, juvenile fish were collected and vitellogenin (Vtg) was measured in whole body homogenates. Native and SDS electrophoresis followed by Western blotting were performed for Vtg identification, and a non-competitive ELISA was developed. In the DES exposure groups (0.5 and 5 mug/l DES), Vtg appeared after 4 days, increased significantly after 8 days and reached a maximum on day 13. Further, a significant increase in the hepatosomatic index (HSI) was found in the 5 mug/l DES exposure group after 21 days. These results indicate that rare minnow provides a good model for assessing endocrine disruption by environmental estrogens. (C) 2004 Elsevier Inc. All rights reserved.

Induction of apoptosis in a carp leucocyte cell line infected with turbot (Scophthalmus maximus L.) rhabdovirus

A rhabdovirus was observed from the diseased turbot (Scophthalmus maximus L.) with lethal syndrome. In this study, a carp leucocyte (CLC) cell line was used to investigate the infection process and cell death mechanism occurring during the virus infection. Strong cytopathogenic effect (CPE) and the morphological changes, such as extreme chromatin condensation, nucleus fragmentation, and apoptotic body formation, were observed under fluorescence microscopy after DAPI staining in the infected CLC cells. Transmission electron microscopy analysis showed cell shrinkage, plasma membrane blebbing, cytoplasm vacuolization, chromatin condensation, nuclear breakdown and formation of discrete apoptotic bodies. The bullet-shaped nucleocapsids were measured and ranged in size from 110 to 150 nm in length and 40 to 60 nm in diameter. And therefore the virus is called Scophthalmus maximus rhabdovirus (SMRV). Agarose gel electrophoresis analysis of the DNA extracted from infected cells showed typical DNA ladder in the course of SMRV infection. Flow cytometry analysis of SMRV infected CLC cells detected apoptotic peak in the virus infected CLC cells. Virus titre analysis and electron microscopic observation revealed that the virus replication fastigium was earlier than that of the apoptosis occurrence. No apoptosis was observed in the CLC infected with UV-inactivated SMRV. All these supported that SMRV infected CLC cells undergo apoptosis and the virus replication is necessary for apoptosis induction of CLC cells. (C) 2004 Published by Elsevier B.V.

Mitochondrial d-loop sequence variation and phylogeny of gobiobotine fishes

The mitochondrial DNA control region is amplified and sequenced from 8 genera and 10 species of gobiobotine fishes. The phylogenetic tree of Gobiobotinae and some representative species of other Cyprinid subfamilies obtained by the method of neighborhood joining, maximum likelihood and maximum parsimony with Danio rerio as an outgroup indicates that Gobiobotinae fishes are a monophyletic group which is close to Gobioninae subfamily. Gobiobotinae should be included into subfamily Gobioninae in terms of phylogenetic analysis. The research result supports that Gobiobotinae can be divided into genus Xenophysogobio and Gobiobotia. Xenophysogabio is the most primitive genera in the subfamily.

Degenerate four-wave mixing based all-optical wavelength conversion in a semiconductor optical amplifier and highly-nonlinear photonic crystal fiber parametric loop mirror

The idler is separated from the co-propagating pump in a degenerate four-wave mixing (DFWM) with a symmetrical parametric loop mirror (PALM), which is composed of two identical SOAs and a 70 m highly-nonlinear photonic crystal fiber (HN-PCF). The signal and pump are coupled into the symmetrical PALM from different ports, respectively. After the DFWM based wavelength conversion (WC) in the clockwise and anticlockwise, the idler exits from the signal port, while the pump outputs from its input port. Therefore, the pump is effectively suppressed in the idler channel without a high-speed tunable filter. Contrast to a traditional PALM, the DFWM based conversion efficiency is increased greatly, and the functions of the amplification and the WC are integrated in the smart SOA and HN-PCF PALM. (C) 2008 Elsevier B.V. All rights reserved.

Pump-Suppressed Nondegenerate Four-Wave Mixing in a Highly Nonlinear Photonic Crystal Fiber Sagnac Loop

We propose a configuration for suppressing pumps in a broad- and flat-hand tunable nondegenerate four-wave mixing (FWM) wavelength converter. The signal and pumps are coupled into a highly nonlinear photonic crystal fiber symmetrical Sagnac loop. After the FWM wavelength conversion in the loop, the idler is separated from the pumps without a filter. In our experiment, a flat wavelength conversion bandwidth of 36 rim, conversion efficiency of-11 dB., pump-to-signal suppression ratio of 48 dB, and idler-to-pump suppression ratio of 15 dB are achieved.

A novel hybrid phase-locked-loop frequency synthesizer using single-electron devices and CMOS transistors

This paper proposes a novel phase-locked loop (PLL) frequency synthesizer using single-electron devices (SEDs) and metal-oxide-semiconductor (MOS) field-effect transistors. The PLL frequency synthesizer mainly consists of a single-electron transistor (SET)/MOS hybrid voltage-controlled oscillator circuit, a single-electron (SE) turnstile/MOS hybrid phase-frequency detector (PFD) circuit and a SE turnstile/MOS hybrid frequency divider. The phase-frequency detection and frequency-division functions are realized by manipulating the single electrons. We propose a SPICE model to describe the behavior of the MOSFET-based SE turnstile. The authors simulate the performance of the PILL block circuits and the whole PLL synthesizer. Simulation results indicated that the circuit can well perform the operation of the PLL frequency synthesizer at room temperature. The PILL synthesizer is very compact. The total number of the transistors is less than 50. The power dissipation of the proposed PLL circuit is less than 3 uW. The authors discuss the effect of fabrication tolerance, the effect of background charge and the SE transfer accuracy on the performance of the PLL circuit. A technique to compensate parameter dispersions of SEDs is proposed.

A novel fast lock-in phase-locked loop frequency synthesizer with direct frequency presetting circuit

This paper proposes a novel, fast lock-in, phase-locked loop (PLL) frequency synthesizer. The synthesizer includes a novel mixed-signal voltage-controlled oscillator (VCO) with a direct frequency presetting circuit. The frequency presetting circuit can greatly speed up the lock-in process by accurately the presetting oscillation frequency of the VCO. We fully integrated the synthesizer in standard 0.35 mu m, 3.3 V complementary metal-oxide-semiconductors (CMOS) process. The entire chip area is only 0.4 mm(2). The measured results demonstrate that the synthesizer can speed up the lock-in process significantly and the lock-in time is less than 10 mu s over the entire oscillation frequency range. The measured phase noise of the synthesizer is -85 dBc/Hz at 10 kHz offset. The synthesizer avoids the tradeoff between the lock-in speed and the phase noise/spurs. The synthesizer monitors the chip temperature and automatically compensates for the variation in frequency with temperature.

Fast Locking and High Accurate Current Matching Phase-Locked Loop

In this paper, a charge-pump based phase-locked loop (CPLL) that can achieve fast locking and tiny deviation is proposed and analyzed. A lock-aid circuit is added to achieve fast locking of the CPLL. Besides, a novel differential charge pump which has good current matching characteristics and a PFD with delay cell has been used in this PLL. The proposed PILL circuit is designed based on the 0.35um 2P4M CMOS process with 3.3V/5V supply voltage. HSPICE simulation shows that the lock time of the proposed CPLL can be reduced by over 72% in comparison to the conventional PILL and its charge pump sink and source current mismatch is only 0.008%.

A Fast-Locking Phase-Locked Loop Using a Seven-State Phase Frequency Detector

A seven-state phase frequency detector (S.S PFD) is proposed for fast-locking charge pump based phase-locked loops (CPPLLs) in this paper. The locking time of the PLL can be significantly reduced by using the seven-state PFD to inject more current into the loop filter. In this stage, the bandwidth of the PLL is increased or decreased to track the phase difference of the reference signal and the feedback signal. The proposed architecture is realized in a standard 0.35 mu m 2P4M CMOS process with a 3.3V supply voltage. The locking time of the proposed PLL is 1.102 mu s compared with the 2.347 mu s of the PLL based on continuous-time PFD and the 3.298 mu s of the PLL based on the pass-transistor tri-state PFD. There are 53.05% and 66.59% reductions of the locking time. The simulation results and the comparison with other PLLs demonstrate that the proposed seven-state PFD is effective to reduce locking time.

A novel fast lock-in phase-locked loop frequency synthesizer with direct frequency presetting circuit

This paper proposes a novel, fast lock-in, phase-locked loop (PLL) frequency synthesizer. The synthesizer includes a novel mixed-signal voltage-controlled oscillator (VCO) with a direct frequency presetting circuit. The frequency presetting circuit can greatly speed up the lock-in process by accurately the presetting oscillation frequency of the VCO. We fully integrated the synthesizer in standard 0.35 mu m, 3.3 V complementary metal-oxide-semiconductors (CMOS) process. The entire chip area is only 0.4 mm(2). The measured results demonstrate that the synthesizer can speed up the lock-in process significantly and the lock-in time is less than 10 mu s over the entire oscillation frequency range. The measured phase noise of the synthesizer is -85 dBc/Hz at 10 kHz offset. The synthesizer avoids the tradeoff between the lock-in speed and the phase noise/spurs. The synthesizer monitors the chip temperature and automatically compensates for the variation in frequency with temperature.