Establishment, characterization, and virus susceptibility of a new marine cell line from red spotted grouper (Epinephelus akaara)

A marine fish cell line from the snout of red spotted grouper Epinephelus akaara, a protogynous hermaphrodite, was established, characterized, and subcultured with more than 60 passages. The grouper snout cell line (GSC) cells multiplied well in Dulbecco's modified Eagle's medium (DMEM) medium supplemented with 10% fetal bovine serum. The optimal growth temperature was 25 degrees C, and morphologically the cells were fibroblastic. Chromosome analysis revealed that the GSC cell line has a normal diploid karyotype with 2n = 8st + 40t. A virus titration study indicated that the cells were susceptible to turbot Scophthalmus Maximus rhabdovirus (SMRV) (10(8.5) TCID50 ml(-1)), while the viral titer of frog Rana grylio virus 9807 (RGV(9807)) reached 10(3.5) TCID50 ml-1. The infection was confirmed by cytopathic effect (CPE), immunofluorescence, and electron microscopy experiments, which detected the viral particles in the cytoplasm of virus-infected cells, respectively. Further, significant fluorescent signals were observed when the GSC cells were transfected with pEGFP vector DNA, indicating their potential utility for transgenic and genetic manipulation studies.

Effects of brominated flame retardants and brominated dioxins on steroidogenesis in H295R human adrenocortical carcinoma cell line

Brominated flame retardants (BFRs) and brominated dioxins are emerging persistent organic pollutants that are ubiquitous in the environment and can be accumulated by wildlife and humans. These chemicals can disturb endocrine function. Recent studies have demonstrated that one of the mechanisms of endocrine disruption by chemicals is modulation of steroidogenic gene expression or enzyme activities. In this study, an in vitro assay based on the H295R human adrenocortical carcinoma cell line, which possesses most key genes or enzymes involved in steroidogenesis, was used to examine the effects of five bromophenols, two polybrominated biphenyls (PBBs 77 and 169), 2,3,7,8-tetrabromodibenzo-p-dioxin, and 2,3,7,8-tetrabromodibenzofuran on the expression of 10 key steroidogenic genes. The H295R cells were exposed to various BFR concentrations for 48 h, and the expression of specific genescytochrome P450 (CYP11A, CYP11B2, CYP17, CYP19, and CYP21), 3 beta-hydroxysteroid dehydrogenase (3PHSD2), 17 beta-hydroxysteroid dehydrogenase (17 beta HSD1 and 17 beta HSD4), steroidogenic acute regulatory protein (StAR), and 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR)-was quantitatively measured using real-time polymerase chain reaction. Cell viability was not affected at the doses tested. Most of the genes were either up- or down-regulated, to some extent, by BFR exposure. Among the genes tested, 3PHSD2 was the most markedly up-regulated, with a range of magnitude from 1.6- to 20-fold. The results demonstrate that bromophenol, bromobiphenyls, and bromodibenzo-p-dioxin/furan are able to modulate steroidogenic gene expression, which may lead to endocrine disruption.

Inductive expression and characterization analysis of Paralichthys olivaceus pigment epithelium-derived factor in a virally infected cell line

Pigment epithelium-derived factor (PEDF) is acknowledged to be a non-inhibitory member of the serine protease inhibitor (serpin) superfamily, with antiangiogenesis, and neuroprotective and immumoregulatory function, mainly in the tissues of nervous system. Here, A PEDF gene homolog, Paralichthys olivaceus PEDF (PoPEDF), was isolated from flounder embryonic cells (FEC) treated with UV-inactivated Grass carp hemorrhage virus (GCHV) and subsequently identified as a differentially expressed gene. The full length of PoPEDF cDNA is 1803 bp with an open reading frame of 1212 bp encoding a 403-amino-acid protein. This deduced protein contains an N-terminal signal peptide, a glycosylation site, a consensus serpin motif, and a 34-mer and a 44-mer fragment, all of which are very conserved in the PEDF family. PoPEDF gene exhibits a conserved exon-intron arrangement with 8 exons and 7 introns. This conserved evolutionary relationship was further confirmed by a phylogenetic analysis, where fish PEDFs and mammalian members formed a well-supported clade. Constitutive expression of PoPEDF was widely detected in many tissues. In response to UV-inactivated GCHV or poly(I:C), PEDF mRNA was upregulated in FEC cells with time. This is the first report on the transcriptional induction of PEDF in virally infected cells. (C) 2005 Elsevier Inc. All rights reserved.

Establishment of a normal medakafish spermatogonial cell line capable of sperm production in vitro

Spermatogonia are the male germ stem cells that continuously produce sperm for the next generation. Spermatogenesis is a complicated process that proceeds through mitotic phase of stem cell renewal and differentiation, meiotic phase, and postmeiotic phase of spermiogenesis. Full recapitulation of spermatogenesis in vitro has been impossible, as generation of normal spermatogonial stem cell lines without immortalization and production of motile sperm from these cells after long-term culture have not been achieved. Here we report the derivation of a normal spermatogonial cell line from a mature medakafish testis without immortalization. After 140 passages during 2 years of culture, this cell line retains stable but growth factor-dependent proliferation, a diploid karyotype, and the phenotype and gene expression pattern of spermatogonial stem cells. Furthermore, we show that this cell line can undergo meiosis and spermiogenesis to generate motile sperm. Therefore, the ability of continuous proliferation and sperm production in culture is an intrinsic property of medaka spermatogonial stem cells, and immortalization apparently is not necessary to derive male germ cell cultures. Our findings and cell line will offer a unique opportunity to study and recapitulate spermatogenesis in vitro and to develop approaches for germ-line transmission.

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.

High efficiency operation of butt joint line-defect-waveguide microlaser in two-dimensional photonic crystal slab

Butt joint line-defect-waveguide microlasers are demonstrated on photonic crystal slabs with airholes in a triangular lattice. Such microlaser is designed to increase the output power from the waveguide edge directly. The output power is remarkably enhanced to 214 times higher by introducing chirped structure in the output waveguide. The lasing mode operates in the linear dispersion region of the output waveguide so that the absorption loss due to the band-edge effect is reduced. The laser resonance is illustrated theoretically using the finite difference time domain method. A practical high power efficiency of 20% is obtained in this microlaser. (C) 2008 American Institute of Physics.

Lifetime distribution of spontaneous emission from line antennas in two-dimensional quasi-periodic photonic crystals

We investigate the lifetime distribution functions of spontaneous emission from line antennas embedded in finite-size two-dimensional 12-fold quasi-periodic photonic crystals. Our calculations indicate that two-dimensional quasi-periodic crystals lead to the coexistence of both accelerated and inhibited decay processes. The decay behaviors of line antennas are drastically changed as the locations of the antennas are varied from the center to the edge in quasi-periodic photonic crystals and the location of transition frequency is varied.

Narrow line-width resonant cavity enhanced photodetectors operating at 1.55 mu m

A method for fabrication of long-wavelength narrow line-width InGaAs resonant cavity enhanced (RCE) photodetectors in a silicon substrate operating at the wavelength range of 1.3-1.6 mu m has been developed. A full width at half maximum (FWHM) of 0.7 nm and a peak responsivity of 0. 16 A/W at the resonance wavelength of 1.55 mu m have been accomplished by using a thick InP layer as part of the resonant cavity. The effects of roughness and tilt of the InP layer surface, and its free carrier absorption, as well as the thickness deviation of the mirror pair on the resonance wavelength shift and the peak quantum efficiency of the RCE photodetectors are analyzed in detail, and approaches for minimizing them toward superior performance are suggested. (C) 2007 Elsevier B.V. All rights reserved.

Formation of self-assembly and the mechanism of Si nanoquantum dots prepared by low pressure chemical vapor deposition

Si nanoquantum dots have been formed by self-assembled growth on the both Si-O-Si and Si-OH bonds terminated SiO2 surfaces using the low-pressure chemical vapor deposition (LPCVD) and surface thermal decomposition of pure SiH4 gas. We have experimentally studied the variation of Si. dot density with Si-OH bonds density, deposition temperature and SiH4 pressure, and analyzed qualitatively the formation mechanism of the Si nanoquantum dots based on LPCVD surface thermal dynamics principle. The results are very. important for the control of the density and size of Si nanoquantum dots, and have potential applications in the new quantum devices.

Electron transport through hierarchical self-assembly of GaAs/AlxGa1-xAs quantum dots

The transmission of electrons through a hierarchical self-assembly of GaAs/AlxGa(1-)xAs quantum dots (QDs) is calculated using the coupled-channel recursion method. Our results reveal that the number of conductance peaks does not change when the barrier widths change, but the intensities decrease as the barrier widths increase. The conductance peaks will shift towards low Fermi energies as the transverse width of GaAs QD increases, as the thickness of GaAs quantum well increases, or as the height of GaAs QDs decreases. Our calculated results may be useful in the application of QDs to photoelectric devices. (c) 2005 American Institute of Physics.

Asymmetric quantum-confined Stark effects of hierarchical self-assembly of GaAs/AlxGa1-xAs quantum dots

Quantum-confined Stark effects in GaAs/AlxGa1-xAs self-assembled quantum dots are investigated theoretically in the framework of effective-mass envelope function theory. The electron and hole energy levels and optical transition energies are calculated in the presence of an electric field in different directions. In our calculation, the effect of finite offset, valence-band mixing, the effects due to the different effective masses of electrons and holes in different regions, and the real quantum dot structures are all taken into account. The results show that the electron and hole energy levels and the optical transition energies can cause blueshifts when the electric field is applied along the opposite to the growth direction. Our calculated results are useful for the application of hierarchical self-assembly of GaAs/AlxGa1-xAs quantum dots to photoelectric devices. (c) 2005 American Institute of Physics.

Effective-mass theory for hierarchical self-assembly of GaAs/AlxGa1-xAs quantum dots

The electronic structures in the hierarchical self-assembly of GaAs/AlxGa1-xAs quantum dots are investigated theoretically in the framework of effective-mass envelope function theory. The electron and hole energy levels and optical transition energies are calculated. In our calculation, the effect of finite offset, valence-band mixing, the effects due to the different effective masses of electrons and holes in different regions, and the real quantum dot structures are all taken into account. The results show that (1) electronic energy levels decrease monotonically, and the energy difference between the energy levels increases as the GaAs quantum dot (QD) height increases; (2) strong state mixing is found between the different energy levels as the GaAs QD width changes; (3) the hole energy levels decrease more quickly than those of the electrons as the GaAs QD size increases; (4) in excited states, the hole energy levels are closer to each other than the electron ones; (5) the first heavy- and light-hole transition energies are very close. Our theoretical results agree well with the available experimental data. Our calculated results are useful for the application of the hierarchical self-assembly of GaAs/AlxGa1-xAs quantum dots to photoelectric devices.

Spontaneous emission lifetime distribution of infinite line antennas in two-dimensional photonic crystals with finite size

The authors calculate the lifetime distribution functions of spontaneous emission from infinite line antennas embedded in two-dimensional disordered photonic crystals with finite size. The calculations indicate the coexistence of both accelerated and inhibited decay processes in disordered photonic crystals with finite size. The decay behavior of the spontaneous emission from infinite line antennas changes significantly by varying factors such as the line antennas' positions in the disordered photonic crystal, the shape of the crystal, the filling fraction, and the dielectric constant. Moreover, the authors analyze the effect of the degree of disorder on spontaneous emission. (c) 2007 American Institute of Physics.

Measurement of a reciprocal four-port transmission line structure using the 16-term error model

A new method to measure reciprocal four-port structures, using a 16-term error model, is presented. The measurement is based on 5 two-port calibration standards connected to two of the ports, while the network analyzer is connected to the two remaining ports. Least-squares-fit data reduction techniques are used to lower error sensitivity. The effect of connectors is deembedded using closed-form equations. (C) 2007 Wiley Periodicals, Inc.

Structure tuning of line-defect waveguides based on silicon-on-insulator photonic crystal slabs

We present fabrication and experimental measurement of a series of photonic crystal waveguides. The complete devices consist of an injector taper down from 3 mu m into a triangular-lattice air-hole single-line-defect waveguide with lattice constant from 410nm to 470nm and normalized radius 0.31. We fabricate these devices on a siliconon-insulator substrate and characterize them using a tunable laser source over a wavelength range from 1510nm to 1640nm. A sharp attenuation at photonic crystal waveguide mode edge is observed for most structures. The edge of guided band is shifted about 30nm with the 10nm increase of the lattice constant. We obtain high-efficiency light propagation and broad flat spectrum response of the photonic crystal waveguides.