An ELISA-like time-resolved fluorescence immunoassay for microcystin detection

Background: A time-resolved fluorescence immunoassay (TRFIA), based on anti-microcystin-LR (MCLR) monoclonal antibodies (MAbs) and europium-labeled antimouse IgG conjugate, was first developed for microcystin detection. Methods: Anti-MCLR MAbs were prepared by a standard method, and the attained MAbs showed a good cross reactivity with MCLR, MCRR and MCYR. The TRFIA was performed in an indirect competitive mode. The detection method of TRFIA was compared with indirect competitive enzyme-linked immunosorbent assay (ELISA) and high-performance liquid chromatography (HPLC). Results: The TRFIA exhibited a typical sigmoidal response for MCLR at concentrations of 0.005-50 ng/ml, with a wide quantitative range between 0.01 and 10 ng/ml, indicating the broadest detective range and the most sensitive of all the methods for microcystins (MCs) detection. Additionally, the TRFIA maintained good reliability through its quantitative range, as evidenced by low coefficients of variation (1.6-12.2%). The toxin data of algal samples assayed from TRFIA were in the same range as those with ELISA and HPLC, implying that the method was reliable and practical for the detection of MCs. Conclusions: The TRFIA may offer a valuable alternative or a substitute for conventional ELISA for microcystin detection. (C) 2004 Elsevier B.V. All rights reserved.

Finite-difference time-domain analysis of deformed square cavity filters with a traveling-wave-like filtering response by mode coupling

An add-drop filter based on a perfect square resonator can realize a maximum of only 25% power dropping because the confined modes are standing-wave modes. By means of mode coupling between two modes with inverse symmetry properties, a traveling-wave-like filtering response is obtained in a two-dimensional single square cavity filter with cut or circular corners by finite-difference time-domain simulation. The optimized deformation parameters for an add-drop filter can be accurately predicted as the overlapping point of the two coupling modes in an isolated deformed square cavity. More than 80% power dropping can be obtained in a deformed square cavity filter with a side length of 3.01 mu m. The free spectral region is decided by the mode spacing between modes, with the sum of the mode indices differing by 1. (c) 2007 Optical Society of America.

Relaxation time and conductivity of comb-like gel polymer electrolytes

Using the copolymer of acrylonitrile (AN), methyl methacrylate (MMA), and poly(ethylene glycol) methyl ether methacrylate as a backbone and poly(ethylene glycol) methyl ether (PEGME) with 1100 molecular weight as side chains, comb-like gel polymers and their Li salt complexes were synthesized. The dynamic mechanical properties and conductivities were investigated. Results showed that the gel copolymer electrolytes possess two glass transitions: alpha-transition and beta-transition. Based on the time-temperature equivalence principle, a master curve was constructed by selecting T. as reference temperature. By reference to T-0 = 50 degrees C, the relation between log c, and c was found to be linear. The master curves are displaced progressively to higher frequencies as the content of plasticizer is increased. The relation between log tau(p) and the content of plasticizer is also linear.

“Deborah Numbers”, Coupling Multiple Space and Time Scales and Governing Damage Evolution to Failure

Two different spatial levels are involved concerning damage accumulation to eventual failure. nucleation and growth rates of microdamage nN* and V*. It is found that the trans-scale length ratio c*/L does not directly affect the process. Instead, two independent dimensionless numbers: the trans-scale one * * ( V*)including the * **5 * N c V including mesoscopic parameters only, play the key role in the process of damage accumulation to failure. The above implies that there are three time scales involved in the process: the macroscopic imposed time scale tim = /a and two meso-scopic time scales, nucleation and growth of damage, (* *4) N N t =1 n c and tV=c*/V*. Clearly, the dimensionless number De*=tV/tim refers to the ratio of microdamage growth time scale over the macroscopically imposed time scale. So, analogous to the definition of Deborah number as the ratio of relaxation time over external one in rheology. Let De be the imposed Deborah number while De represents the competition and coupling between the microdamage growth and the macroscopically imposed wave loading. In stress-wave induced tensile failure (spallation) De* < 1, this means that microdamage has enough time to grow during the macroscopic wave loading. Thus, the microdamage growth appears to be the predominate mechanism governing the failure. Moreover, the dimensionless number D* = tV/tN characterizes the ratio of two intrinsic mesoscopic time scales: growth over nucleation. Similarly let D be the “intrinsic Deborah number”. Both time scales are relevant to intrinsic relaxation rather than imposed one. Furthermore, the intrinsic Deborah number D* implies a certain characteristic damage. In particular, it is derived that D* is a proper indicator of macroscopic critical damage to damage localization, like D* ∼ (10–3~10–2) in spallation. More importantly, we found that this small intrinsic Deborah number D* indicates the energy partition of microdamage dissipation over bulk plastic work. This explains why spallation can not be formulated by macroscopic energy criterion and must be treated by multi-scale analysis.

RNAi suppression of zebrafish peptidoglycan recognition protein 6 (zfPGRP6) mediated differentially expressed genes involved in Toll-like receptor signaling pathway and caused increased susceptibility to Flavobacterium columnare

In Drosophila, Toll signaling cascade, which resembles the mammalian Toll-like receptor (TLR)/IL-1R signaling pathways and regulates the expression of anti-microbial peptide genes, mainly relies on peptidoglycan recognition proteins (PGRPs) for the detection of bacterial pathogens. To explore the effect of zebrafish peptidoglycan recognition protein 6 (zfPGRP6) on Toll-like receptor signaling pathway, RNA interference (siRNA) and real time quantitative PCR (RQ-PCR) methods were used to identify differentially expressed genes regulated by zfPGRP6. The target genes included TLR2, TLR3, TLR5, TLR7, TLR8, IL1R, Sterile-alpha and Armadillo motif containing protein (SARM), myeloid differentiation factor 88 (MyD88) and nuclear factor (NF)-kappa B2 (p100/p52). The results of RQ-PCR showed that RNAi-mediated Suppression of zfPGRP6 significantly down-regulated the expression of TLR2, TLR5, IL1R, SARM, MyD88 and p100/p52. The expression of beta-defensin-1 was also down-regulated in those embryos silenced by zfPGRP6. In challenge experiments to determine the anti-bacterial response to Gram-negative bacteria, RNAi knock-down of zfPGRP6 markedly increased susceptibility to Flavobacterium columnare. (C) 2008 Elsevier B.V. All rights reserved.

C1q-like inhibits p53-mediated apoptosis and controls normal hernatopoiesis during zebrafish embryogenesis

Except for the complement C1q, the immunological functions of other C1q family members have remained unclear. Here we describe zebrafish C1q-like, whose transcription and translation display a uniform distribution in early embryos, and are restricted to mid-hind brain and eye in later embryos. In vitro studies showed that C1q-like could inhibit the apoptosis induced by ActD and CHX in EPC cells, through repressing caspase 3/9 activities. Moreover, its physiological roles were studied by morpholino-mediated knockdown in zebrafish embryogenesis. In comparison with control embryos, the C1q-like knockdown embryos display obvious defects in the head and cramofacial development mediated through p53-induced apoptosis, which was confirmed by the in vitro transcribed C1q-like mRNA or p53 MO co-injection. TUNEL assays revealed extensive cell death, and caspase 3/9 activity measurement also revealed about two folds increase in C1q-like morphant embryos, which was inhibited by p53 MO co-injection. Real-time quantitative PCR showed the up-regulation expression of several apoptosis regulators such as p53, mdm2, p21, Box and caspase 3, and down-regulation expression of hbae1 in the C1q-like morphant embryos. Knockdown of C1q-like in zebrafish embryos decreased hemoglobin production and impaired the organization of mesencephalic vein and other brain blood vessels. Interestingly, exposure of zebrafish embryos to UV resulted in an increase in mRNA expression of C1q-like, whereas over-expression of C1q-like was not enough resist to the damage. Furthermore, C1q-like transcription was up-regulated in response to pathogen Aeromonas hydrophila, and embryo survival significantly decreased in the C1q-like morphants after exposure to the bacteria. The data suggested that C1q-like might play an antiapoptotic and protective role in inhibiting p53-dependent and caspase 3/9-mediated apoptosis during embryogenesis, especially in the brain development, and C1q-like should be a novel regulator of cell survival during zebrafish embryogenesis. (c) 2008 Elsevier Inc. All rights reserved.

Toll-like receptor 3 regulates Mx expression in rare minnow Gobiocypris rarus after viral infection

Toll-like receptor 3 (TLR3) plays a key role in activating immune responses during viral infection. To study the genes involved in the regulatory function of TLR3 in the rare minnow Gobiocypris rarus after viral infection, a full-length cDNA of TLR3 (GrTLR3) with a splice variant (GrTLR3s) was identified by homologous cloning and RACE techniques. The antiviral effector molecule Mx gene was cloned and partially sequenced. The mRNA expression levels of GrTLR3, GrTLR3s, and Mx were studied in different tissues before and after virus infection by real-time quantitative RT-PCR. The transcripts of all three genes in liver were significantly increased following GCRV infection (P<0.05). The mRNA levels in liver were upregulated at 24 h post-injection for GrTLR3 and GrTLR3s, and at 12 h for Mx. The upregulated expression levels were several folds for GrTLR3s, tens of folds for GrTLR3, and hundreds of folds for Mx. By semi-quantitative RT-PCR, GrTLR3 and Mx expressed at all the developmental stages, whereas GrTLR3s could only be detected at later developmental stages. Using RNAi and transgenic techniques, GrTLR3 mediated Mx expression but GrTLR3s did not. The time-dependent upregulation of receptor and effector, and the Mx over-expression dependent on TLR3, indicated that GrTLR3 regulated Mx expression in viral infection through a configuration change in rare minnow, and its splice variant did not contribute to the process.

Dioxin-like components in human breast milk collected from Hong Kong and Guangzhou

The H4IIE rat hepatoma cell line was employed as a cell model to screen 7-ethoxyresorufin O-deethylase (EROD)-TCDD equivalents (EROD-TEQ) of human breast milk samples collected from Hong Kong and Guangzhou, China. The screening methods employed a 96-well plate spectrofluorometer-EROD assay. For cell-line validation, our results demonstrated a dose-dependent increase in the Ah receptor-mediated response (i.e., CYP1A1 mRNA and EROD) of the cells upon exposure to a number of known Ah receptor agonists, including 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 2,3,7,8-tetrachlorodibenzothiophene, benzo[a]pyrene, and beta-naphthaflavone. TCDD induced CYP1A1 mRNA and EROD was in a close positive correlation (r = 0.98). For the screening of dioxin-like compounds, breast milk samples collected during lactation weeks 3-5 were used. One hundred (from Hong Kong) and 48 (from Guangzhou) breast milk samples were assayed, of which 65% and 68% of the samples, respectively, showed detectable dioxin-like activities using the H4IIE cell EROD screening method. For sixty-five samples from Hong Kong the mean EROD-TEQ values ranged from 58.1 to 96.5 pg/g of milk fat for those aged 21-36 years while 32 samples from Guangzhou had mean values of 98.8-202.1 pg/g of milk fat. In comparisons of the EROD-TEQ values for different age groups from both cities, there were no significant differences (P < 0.05). However, the mean and median EROD-TEQ values of the Guangzhou population were in general higher than those of the Hong Kong population. The results of the present study indicate that it is feasible to use the H4IIE cell-line as a model for screening dioxin-like compounds in human breast milk. In addition, the method is rapid and cost-effective, particularly for a routine and high-throughput sample screening analysis, compared to the costly and time-intensive chemical analytical techniques. (C) 2003 Elsevier Inc. All rights reserved.

Wavelength Coded Optical Time-Domain Reflectometry

This paper presents a wavelength coded optical timedomain reflectometry based on optical heterodyne technique. In this scheme, the probe and reference optical pulses have different wavelengths. This enables optical heterodyne detection to be used to improve the system performances significantly. We demonstrate a spatial resolution of 2.5 m within a range of 60 km in weak-reflection signal detection and direct observation of Brillouin scattering over a long optical fiber, suggesting online fiber sensing possible. The principle of wavelength coding is applicable to other systems like lidar and radar to increase receiver sensitivity and simplify system structure.

Electrical control of dynamic spin splitting induced by exchange interaction as revealed by time-resolved Kerr rotation in a degenerate spin-polarized electron gas

Manipulation of the spin degree of freedom has been demonstrated in a spin-polarized electron plasma in a heterostructure by using exchange-interaction-induced dynamic spin splitting rather than the Rashba and Dresselhaus types, as revealed by time-resolved Kerr rotation. The measured spin splitting increases from 0.256 meV to 0.559 meV as the bias varies from -0.3 V to -0.6 V. Both the sign switch of the Kerr signal and the phase reversal of Larmor precessions have been observed with biases, which all fit into the framework of exchange-interaction-induced spin splitting. The electrical control of it may provide a new effective scheme for manipulating spin-selected transport in spin FET-like devices. Copyright (C) EPLA, 2008.

Spin dynamics in the second subband of a quasi-two-dimensional system studied in a single-barrier heterostructure by time-resolved Kerr rotation

Spin dynamics in the first and second subbands have been examined simultaneously by time resolved Kerr rotation in a single-barrier heterostructure of a 500 nm thick GaAs absorption layer. By scanning the wavelengths of the probe and pump beams towards the short wavelength in the zero magnetic field, the spin coherent time T-2(1)* in the 1st subband E-1 decreases in accordance with the D'yakonov-Perel' (DP) spin decoherence mechanism. Meanwhile, the spin coherence time T-2(2)* in the 2nd subband E-2 remains very low at wavelengths longer than 810 nm, and then is dramatically enhanced afterwards. At 803 nm, T-2(2)* (450 ps) becomes ten times longer than T-2(1)* (50 ps). A new feature has been discovered at the wavelength of 811nm under the bias of -0.3V (807nm under the bias of -0.6V) that the spin coherence times (T-2(1)* and T-2(2)*) and the effective g* factors (vertical bar g*(E-1)vertical bar and vertical bar g*(E-2)vertical bar) all display a sudden change, presumably due to the "resonant" spin exchange coupling between two spin opposite bands. Copyright (C) EPLA, 2008.

Investigation of mode characteristics for a square cavity with a pedestal by a three-dimensional finite-difference time-domain technique

Mode characteristics of a strongly confined square cavity suspended in air via a pedestal on the substrate are investigated by a three-dimensional finite-difference time-domain technique. The mode wavelengths and mode quality factors (Q factors) are calculated as the functions of the size of the pedestal and the slope angle 0 of the sidewalls of the square slab, respectively For the square slab with side length of 2 mu m, thickness of 0.2 mu m, and refractive index of 3.4, on a square pedestal with refractive index of 3.17, the Q factor of the whispering-gallery (WG)-like mode transverse-electric TE(3.5)o first increases with the side length b of the square pedestal and then quickly decreases as b > 0.4 mu m, but the Q factor of the WG-like mode TE(4.6)o drops down quickly as b > 0.2 mu m, owing to their different symmetries. The results indicate that the pedestal can also result in mode selection in the WG-like modes. In addition, the numerical results show that the Q factors decrease 50% as the slope angle of the sidewalls varies from 90 degrees to 80 degrees. The mode characteristics of WG-like modes in the square cavity with a rectangular pedestal are also discussed. The results show that the nonsquare pedestal largely degrades the WG-like modes. (c) 2006 Optical Society of America

WG-like modes selectivity in a square cavity with posts

We investigate the characteristics of Whispering-Gallery(WG)-like modes in a square cavity with posts by employing the two-dimentional (2D) finite-difference time-domain (FDTD) technique combined with the effective index method. The results indicate that the posts can result in mode selection in the WG-like modes. The WG-like modes with odd mode numbers are not much sensitive to the sizes of the posts. However, the quality factor (i.e. Q-factor) of the WG-like modes with even mode numbers decreases sharply with the increasing size of the posts. The decreasing Q-factor is attributed to mode leakage and scattering loss due to the presence of the post. The mode selection increases with the mode spacing of square cavity twice in an optimized strucure.

Ultrafast Kerr rotations and zero-field dephasing time of electron spins in InAs/GaAs quantum disks

Time-resolved Kerr rotation (TRKR) measurements based on pump-probe arrangement were carried out at 5 K on the monolayer fluctuation induced InAs/GaAs quantum disks grown on GaAs substrate without external magnetic field. The lineshape of TRKR signals shows an unusual dependence on the excitation wavelength, especially antisymmetric step-shaped structures appearing when the excitation wavelength was resonantly scanned over the heavy- and light-hole subbands. Moreover, these step structures possess an almost identical decay time of similar to 40 Ps which is believed to be the characteristic spin dephasing time of electrons in the extremely narrow InAs/GaAs quantum disks.

Guiding of 150 keV O6+ ions through nanocapillaries in an uncoated Al2O3 membrane: special time dependence of the transmission profile width

This paper reports that the transmission of O6+ ions with energy of 150keV through capillaries in an uncoated Al2O3 membrane was measured, and agreements with previously reported results in general angular distribution of the transmitted ions and the transmission fractions as a function of the tilt angle well fitted to Gaussian-like functions were observed. Due to using an uncoated capillary membrane, our c is larger than that using a gold-coated one with a smaller value of E-p/q, which suggests a larger equilibrium charge Q(infinity) in our experiment. The observed special width variation with time and a larger width than that using a smaller E-p/q were qualitatively explained by using mean-field classical transport theory based on a classical-trajectory Monte Carlo simulation.