Enzyme-linked immunosorbent assay of changes in serum levels of growth hormone (cGH) in common carps (Cyprinus carpio)

The aim of the present study was to purify the common native carp growth hormone (ncGH), produce monoclonal antibodies (mAbs) to common native carp growth hormone (ncGH), and further enhance the sensitivity of enzyme-linked immunosorbent assays (ELISA) for ncGH. Additionally, we investigated changes in serum ncGH levels in carps raised in different environmental conditions. The recombinant grass carp (Ctenopharyngodon idella) growth hormone was purified and used as antigen to immunize the rabbit. The natural ncGH was isolated from the pituitaries of common carp. SDS-PAGE and Western blot utilizing the polyclonal anti-rgcGH antibody confirmed the purification of ncGH from pituitaries. Purified ncGH was then used as an immunogen in the B lymphocyte hybridoma technique. A total of 14 hybridoma cell lines (FMU-cGH 1-14) were established that were able to stably secrete mAbs against ncGH. Among them, eight clones (FMU-cGH1-6, 12 and 13) were successfully used for Western blot while nine clones (FMU-cGH 1-7, 9 and 10) were used in fluorescent staining and immunohistochemistry. Epitope mapping by competitive ELISA demonstrated that these mAbs recognized five different epitopes. A sensitive sandwich ELISA for detection of ncGH was developed using FMU-cGH12 as the coating mAb and FMU-cGH6 as the enzyme labeled mAb. This detection system was found to be highly stable and sensitive, with detection levels of 70 pg/mL. Additionally, we found that serum ncGH levels in restricted food group and in the net cage group increased 6.9-and 5.8-fold, respectively, when compared to controls, demonstrating differences in the GH stress response in common carp under different living conditions.

Antioxidant enzyme activities of Microcystis aeruginosa in response to nonylphenols and degradation of nonylphenols by M-aeruginosa

The aim of this study was to examine the effects of chemical nonylphenols (NPs) on the antioxidant system of Microcystis aeruginosa strains. The degradation and sorption of NPs by M. aeruginosa were also evaluated. High concentrations of NPs (1 and 2 mg/l) were found to cause increases in superoxidase dismutase (SOD) and glutathione-S-transferase (GST) activities and in glutathione (GSH) levels. These results suggest that toxic stress manifested by elevated SOD and GST levels and GSH contents may be responsible for the toxicity of NPs to M. aeruginosa and that the algal cells could improve their antioxidant and detoxification ability through the enhancement of enzymatic and nonenzymatic prevention substances. The observed elevations in GSH levels and GST activities were relatively higher than those in SOD activities, indicating that GSH and GST contributed more in eliminating toxic effects than SOD. Low concentrations of NPs (0.05-0.2 mg/l) enhanced cell growth and decreased GST activity in algal cells of M. aeruginosa, suggesting that NPs may have acted as a protecting factor, such as an antioxidant. The larger portion of the NPs (> 60%) disappeared after 12 days of incubation, indicating the strong ability of M. aeruginosa to degrade the moderate persistent NP compounds. The sorption ratio of M. aeruginosa after a 12-day exposure to low nominal concentrations of NPs (0.02-0.5 mg/l) was relatively high (> 30%). The fact that M. aeruginosa effectively resisted the toxic effects of NPs and strongly degraded these pollutants indicate that M. aeruginosa cells have a strong ability to adapt to variations in environmental conditions and that low and moderate concentrations of organic compounds may favor its survival. Further studies are needed to provide detailed information on the fate of persistent organic pollutants and the survival of algae and to determine the possible role of organic pollutants in the occurrence of water blooms in eutrophic lakes.

An enzyme-linked immunosorbent assay for rare minnow (Gobiocypris rarus) vitellogenin and comparison of vitellogenin responses in rare minnow and zebrafish (Danio rerio)

A competitive enzyme-linked immunosorbent assay (ELISA) was developed to determine vitellogenin (Vtg) in rare minnow (Gobiocypris rarus) based on the separation and purification of rare minnow Vtg (r-Vtg) as well as the production of polyclonal antibody against r-Vtg in rabbits. Three different ELISAs for measuring r-Vtg were then compared: (1) indirect ELISA with the antibody against carp (Cyprinus carpio) Vtg (c-Vtg) (IC-ELISA); (2) competitive ELISA with the antibody against c-Vtg, and using r-Vtg for coating the plates and preparing standard curve (CC-ELISA); (3) competitive ELISA with the antibody against r-Vtg, and using r-Vtg for coating the plates and preparing standard curve (CR-ELISA). The result showed that the homologous CR-ELISA was the most sensitive among the three assays for quantifying r-Vtg. The sensitivities to 17 alpha-ethinylestradiol (EE2) Of rare minnow and zebrafish (Danio rerio) were compared upon the establishment of homologous competitive ELISA. The lowest observed effect concentrations (LOECs) to induce Vtg were found to be 0.8 ng EE2 l(-1) for rare minnow and 4 ng EE2 l(-1) for zebrafish respectively. Afterwards, CR-ELISA was applied to measure Vtg concentration in whole body homogenate (WBH) of juvenile rare minnow fed by three diets (tubifex from wastewater treatment plant, Artemia nauplii and commercial pellet food), and the agreements between bioassay and GC-MS analysis demonstrated that rare minnow was a sensitive fish model for assessing estrogenic effects of endocrine disrupting compounds in aquatic environment. (c) 2006 Elsevier B.V. All rights reserved.

Effects of age and dietary protein level on digestive enzyme activity and gene expression of Pelteobagrus fulvidraco larvae

The present research studied the effects of age and dietary protein level on pepsin, trypsin and amylase activity and their mRNA level in Petteobagrus fulvidraco larvae from 3 to 26 days after hatch (DAH). Three DAH larvae were fed three isoenergetic diets, containing 42.8% (CP 43), 47.3% (CP 47) and 52.8% (CP 53) crude protein. Live food (newly hatched Artemia, unenriched) was included as a control. The effects of age on enzyme activity and mRNA were as follows: pepsin and trypsin activity in all treatment groups showed a significant (P < 0.05) increase at the beginning and decrease later although the timing of decrease was not the same among treatment groups and between the digestive enzymes. Pepsin and trypsin mRNA level followed the pattern of their respective enzyme changes. Age significantly affected amylase activity (P < 0.05) while age had no effect on amylase mRNA during the experimental period. The four diets significantly (P < 0.05) affected activity and mRNA level of pepsin and trypsin. Diets did not affect amylase activity or mRNA level. These results suggest that the effects of age on pepsin and trypsin gene expressions are at the transcriptional level. Dietary protein level does affect pepsin and trypsin gene expression in the early life of P. fulvidraco. There were no transcriptional effects on amylase gene expression. (c) 2005 Elsevier B.V. All rights reserved.

Gene cloning and functional analysis of glycosaminoglycan-degrading enzyme chondroitin AC lyase from Flavobacterium columnare G(4)

The chondroitin AC lyase gene, cslA, was cloned for the first time from the fish bacterial pathogen F. columnare G(4). From the first transcription initiation site, the cslA extends 2620 nucleotides to the end of the 3' region. The open reading frame of cslA transcript has 2286 nucleotides encoding 762 amino acids with a 16 residues long signal peptide at the N-terminus. The gene, cslA was then successfully expressed in Escherichia coli and recombinant chondroitin AC lyase, rChonAC was purified, with its lytic activity analyzed. Zymography analysis copolymerized with chondroitin sulphate revealed the lytic activity of rChonAC and also the crude native ChonAC isolated from periplamic space of cultured F. columnare G(4). The low level of lytic activity observed in crude native ChonAC may be due possibly to the low level of expression of this gene in the cultured condition. The expression and the role of this virulence factor is of interest for further research on the pathogenesis of F. columnare.

A new enzyme immunoassay for PCDD/F TEQ screening in environmental samples: Comparison to micro-EROD assay and to chemical analysis

A new Enzyme ImmunoAssay (EIA) for PCDD/F TEQ measurement in extracts of environmental samples was described. The bioassay TEQ which derived from EIA and EROD were compared with each other and with results from chemical analysis. For all environmental samples, the EROD-TEQ is higher than the value from chemical analysis. However, the EIA-TEQ is much more identical with the value from chemical analysis. Our results indicate that the EIA assay is a complementary method to the EROD assay and should be useful as a rapid and sensitive screening tool for environmental samples in many situations. (C) 1999 Elsevier Science Ltd. All rights reserved

Hydrogen sensors based on AlGaN/AIN/GaN HEMT

Pt/AlGaN/AIN/GaN high electron mobility transistors (HEMT) were fabricated and characterized for hydrogen sensing. Pt and Ti/Al/Ni/Au metals were evaporated to form the Schottky contact and the ohmic contact, respectively. The sensors can be operated in either the field effect transistor (FET) mode or the Schottky diode mode. Current changes and time dependence of the sensors under the FET and diode modes were compared. When the sensor was operated in the FET mode, the sensor can have larger current change of 8 mA, but its sensitivity is only about 0.2. In the diode mode, the current change was very small under the reverse bias but it increased greatly and gradually saturated at 0.8 mA under the forward bias. The sensor had much higher sensitivity when operated in the diode mode than in the FET mode. The oxygen in the air could accelerate the desorption of the hydrogen and the recovery of the sensor. (c) 2007 Elsevier Ltd. All rights reserved.

Hydrogen sensors based on AlGaN/AIN/GaN Schottky diodes

Pt/AlGaN/AIN/GaN Schottky diodes are fabricated and characterized for hydrogen sensing. The Pt Schottky contact and the Ti/Al/Ni/Au ohmic contact are formed by evaporation. Both the forward and reverse currents of the device increase greatly when exposed to hydrogen gas. A shift of 0.3 V at 300K is obtained at a fixed forward current after switching from N-2 to 10%H-2+N-2. The sensor responses under different concentrations from 50ppm H-2 to 10%H-2+N-2 at 373K are investigated. Time dependences of the device forward current at 0.5 V forward bias in N-2 and air atmosphere at 300 and 373K are compared. Oxygen in air accelerates the desorption of the hydrogen and the recovery of the sensor. Finally, the decrease of the Schottky barrier height and sensitivity of the sensor are calculated.

Development of current-based microscopic defect analysis method using optical filling techniques for the defect study on heavily irradiated high-resistivity Si sensors/detectors

Current-based microscopic defect analysis method such as current deep level transient spectroscopy (I-DLTS) and thermally stimulated current have been developed over the years at Brookhaven National Laboratory (BNL) for the defect characterizations on heavily irradiated (Phi(n) >= 10(13) n/cm(2)) high-resistivity (>= 2 k Omega cm) Si sensors/detectors. The conventional DLTS method using a capacitance transient is not valid on heavily irradiated high-resistivity Si sensors/detectors. A new optical filling method, using lasers with various wavelengths, has been applied, which is more efficient and suitable than the traditional voltage-pulse filling. Optimum defect-filling schemes and conditions have been suggested for heavily irradiated high-resistivity Si sensors/detectors. (c) 2006 Published by Elsevier Ltd.

H+ISFET-based biosensor for determination of penicillin G

A biosensor based on an H+ ion sensitive field effect transistor (H+-ISFET) and penicillin G acylase has been developed. The response time of the sensor to different concentrations of penicillin G was 30 s. In a 20 mM phosphate buffer at pH 7.0, the linear range of the calibration curve was from 0.5 to 8 mM. The coefficients of variation for three samples with 20 repeated measurements were below 5%. Stability of the sensor could reach about 6 months and more than 1000 runs were performed without a significant decrease of the output value. The sensor was tested for measurement of the penicillin G content in penicillin fermentation broth. Forty samples with low and high concentrations of penicillin G were chosen for the correlation test. The values assayed by the sensor method were compared with the values assayed by HPLC method, the correlation coefficient (r) was 0.9944 and the regression equation was y = 1.034X - 2083.7 respectively. The different measuring methods are discussed in the text. (C) 1998 Published by Elsevier Science S.A. All rights reserved.

Hydrogen sensors based on Pt-AlGaN/GaN back-to-back Schottky diode

In this paper, platinum (Pt) with a thickness of 45 nm was sputtered on the surface of AlGaN/GaN heterostructure to form the Schottky contact and the back-to-back Schottky diodes were characterized for H-2 sensing at room temperature. Both the forward and reverse current of the devices increased with exposure to H-2 gas, which was attributed to Schottky barrier height reduction caused by hydrogen absorption in the catalytic metals. A shift of 0.7 V at 297 K was obtained at a fixed forward current of 0.1 mA after switching from N-2 to 40% H-2 in N-2. The sensor's responses under different concentrations from 2500 ppm H-2 to 40% H-2 in N-2 at 297 K were investigated. Time response of the sensor at a fixed bias of 1 V was given. Finally, the decrease of the Schottky barrier height and the sensitivity of the sensor were calculated. (C) 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Hydrogen sensors based on Pt-AlGN/AIN/GaN Schottky diode - art. no. 68291R

Pt/AlGaN/AlN/GaN Schottky diodes have been fabricated and characterized for H-2 sensing. Platinum (Pt) with a thickness of 20nm was evaporated on the sample to form the Schottky contact. The ohmic contact, formed by evaporated Ti/Al/Ni/Au metals, was subsequently annealed by a rapid thermal treatment at 860 degrees C for 30 s in N-2 ambience. Both the forward and reverse current of the device increased greatly when exposed to H-2 gas. The sensor's responses under different hydrogen concentrations from 500ppm to 10% H-2 in N-2 at 300K were investigated. A shift of 0.45V at 297K is obtained at a fixed forward current for switching from N-2 to 10% H-2 in N-2. Time response of the sensor at a fixed bias of 0.5 V was also measured. The turn-on response of the device was rapid, while the recovery of the sensor at N-2 atmosphere was rather slow. But it recovered quickly when the device was exposed to the air. The decrease in the barrier height of the diode was calculated to be about 160meV upon introduction of 10% H-2 into the ambient. The sensitivity of the sensor is also calculated. Some thermodynamics analyses have been done according to the Langmuir isotherm equation.

A Novel Low-Power Digital Baseband Circuit for UHF RFID Tag with Sensors

A novel low-power digital baseband circuit for UHF RFID tag with sensors is presented in this paper. It proposes a novel baseband architecture and a new operating scheme to fulfill the sensor functions and to reduce power consumption. It is also compatible with the EPC C1G2 UHF RFID protocol. It adopts some advanced low power techniques for system design and circuit design: adaptive clock-gating, multi-clock domain and asynchronous circuit. The baseband circuit is implemented in 0.18um 1P3M standard CMOS process. ne chip area is 0.28 mm(2) excluding test pads. Its power consumption is 25uW under 1.1V power supply.

High resolution interrogation technique based on linear photodiode array spectrometer for fiber Bragg grating Sensors - art. no. 65952C

A linear photodiode array spectrometer based, high resolution interrogation technique for fiber Bragg grating sensors is demonstrated. Spline interpolation and Polynomial Approximation Algorithm (PAA) are applied to the data points acquired by the spectrometer to improve the original PAA based interrogation method. Thereby fewer pixels are required to achieve the same resolution as original. Theoretical analysis indicates that if the FWHM of a FBG covers more than 3 pixels, the resolution of central wavelength shift will arrive at less than 1 pm. While the number of pixels increases to 6, the nominal resolution will decrease to 0.001 pm. Experimental result shows that Bragg wavelength resolution of similar to 1 pm is obtained for a FBG with FWHM of similar to 0.2 nm using a spectrometer with a pixel resolution of similar to 70 pm.