972 resultados para double antibody sandwich ELISA
Resumo:
The low speed impact responses of simply-supported and clamped sandwich beams with corrugated and Y-frame cores have been measured in a drop-weight apparatus at 5 m s-1. The AISI 304 stainless steel sandwich beams comprised two identical face sheets and represented 1:20 scale versions of ship hull designs. No significant rate effects were observed at impact speeds representative of ship collisions: the drop-weight responses were comparable to the ones measured quasi-statically. Moreover, the corrugated and Y-frame core beams had similar performances. Three-dimensional finite element (FE) models simulated the experiments and were in good agreement with the measurements. The simulations demonstrated correctly that the sandwich beams collapsed by core indentation under both quasi-static loading and in the drop-weight experiments. These FE models were then used to investigate the sensitivity of impact response to (i) velocity, over a wider range of velocities than achievable with the drop-weight apparatus, and (ii) the presence of the back face sheet. The dynamic responses of sandwich beams with both front and back face sheets were found to be within 20% of the quasi-static responses for speeds less than approximately 5 m s-1. This suggests that quasi-static considerations are adequate to model the collision of a sandwich ship hull. By contrast, beams without a back face collapsed by Brazier buckling under quasi-static loading conditions, and by core indentation at a loading velocity of 5 m s-1. Thus, dynamic considerations are needed in ship hull designs that do not employ a back face. © 2014 Elsevier Ltd. All rights reserved.
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The dynamic response of end-clamped sandwich and monolithic beams of equal areal mass subjected to loading via high-velocity slugs of dry and water-saturated sand is measured using a novel laboratory-based method. The sandwich beams comprise aluminium face sheets and an aluminium honeycomb core: the effect of sandwich core strength and beam thickness on the dynamic beam deflection is investigated by varying the orientation and height of the anisotropic aluminium honeycomb core material. High-speed imaging is used to measure the transient transverse deflection of the beams and to record the dynamic modes of deformation. The measurements show that sandwich beams with thick, strong cores are optimal and that these beams significantly outperform monolithic beams of equal mass. The water-saturated sand slugs cause significantly higher deflections compared to the dry sand slugs having the same mean slug velocity and we demonstrate that this enhanced deflection is due to the larger mass of the water-saturated slugs. Finally, we show that the impact of sand slugs is equivalent to the impact of a crushable foam projectile. The experiments using foam projectiles are significantly simpler to perform and thus represent a more convenient laboratory technique. © 2014 Elsevier Ltd. All rights reserved.
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Photosynthesis by phytoplankton cells in aquatic environments contributes to more than 40% of the global primary production (Behrenfeld et al., 2006). Within the euphotic zone (down to 1% of surface photosynthetically active radiation [PAR]), cells are exposed not only to PAR (400-700 nm) but also to UV radiation (UVR; 280-400 nm) that can penetrate to considerable depths (Hargreaves, 2003). In contrast to PAR, which is energizing to photosynthesis, UVR is usually regarded as a stressor (Hader, 2003) and suggested to affect CO2-concentrating mechanisms in phytoplankton (Beardall et al., 2002). Solar UVR is known to reduce photosynthetic rates (Steemann Nielsen, 1964; Helbling et al., 2003), and damage cellular components such as D1 proteins (Sass et al., 1997) and DNA molecules (Buma et al., 2003). It can also decrease the growth (Villafane et al., 2003) and alter the rate of nutrient uptake (Fauchot et al., 2000) and the fatty acid composition (Goes et al., 1994) of phytoplankton. Recently, it has been found that natural levels of UVR can alter the morphology of the cyanobacterium Arthrospira (Spirulina) platensis (Wu et al., 2005b). On the other hand, positive effects of UVR, especially of UV- A (315-400 nm), have also been reported. UV- A enhances carbon fixation of phytoplankton under reduced (Nilawati et al., 1997; Barbieri et al., 2002) or fast-fluctuating (Helbling et al., 2003) solar irradiance and allows photorepair of UV- B-induced DNA damage (Buma et al., 2003). Furthermore, the presence of UV-A resulted in higher biomass production of A. platensis as compared to that under PAR alone (Wu et al., 2005a). Energy of UVR absorbed by the diatom Pseudo-nitzschia multiseries was found to cause fluorescence (Orellana et al., 2004). In addition, fluorescent pigments in corals and their algal symbiont are known to absorb UVR and play positive roles for the symbiotic photosynthesis and photoprotection (Schlichter et al., 1986; Salih et al., 2000). However, despite the positive effects that solar UVR may have on aquatic photosynthetic organisms, there is no direct evidence to what extent and howUVR per se is utilized by phytoplankton. In addition, estimations of aquatic biological production have been carried out in incubations considering only PAR (i. e. using UV-opaque vials made of glass or polycarbonate; Donk et al., 2001) without UVR being considered (Hein and Sand-Jensen, 1997; Schippers and Lurling, 2004). Here, we have found that UVR can act as an additional source of energy for photosynthesis in tropical marine phytoplankton, though it occasionally causes photoinhibition at high PAR levels. While UVR is usually thought of as damaging, our results indicate that UVR can enhance primary production of phytoplankton. Therefore, oceanic carbon fixation estimates may be underestimated by a large percentage if UVR is not taken into account.
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In a previous study, a scFv phage display library against white spot syndrome virus (WSSV) was constructed and yielded a clone designated A I with conformational specificity against native but not denatured viral antigen. Although the clone A1 has been used successfully as a diagnostic antibody, its precise target antigen has not been elucidated. A different strategy was adopted involving the construction of a second T7 phage display library utilizing mRNA isolated from shrimp infected with WSSV. Following RT-PCR and T7 phage library construction, phages displaying the candidate epitope were selected with A I scFv. Since successive enrichment steps were not associated with an increased titer of the phages, enrichment after successive tests was confirmed by PCR resulting in the prefer-red selection of a specific DNA sequence encoding a novel nucleocapsid protein WSSV388. Immune electron microscopy revealed that WSSV388 is located on the nucleocapsid. This result demonstrated that unknown antigen could be identified by phage display using the epitope conformation dependent scFv. (c) 2006 Elsevier B.V. All rights reserved.
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We demonstrate wide-band ultrafast optical pulse generation at 1, 1.5, and 2 μm using a single-polymer composite saturable absorber based on double-wall carbon nanotubes (DWNTs). The freestanding optical quality polymer composite is prepared from nanotubes dispersed in water with poly(vinyl alcohol) as the host matrix. The composite is then integrated into ytterbium-, erbium-, and thulium-doped fiber laser cavities. Using this single DWNT-polymer composite, we achieve 4.85 ps, 532 fs, and 1.6 ps mode-locked pulses at 1066, 1559, and 1883 nm, respectively, highlighting the potential of DWNTs for wide-band ultrafast photonics.
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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.
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Production of zebrafish by modifying endogenous growth hormone (GH) gene through homologous recombination is described here. We first constructed the targeting vectors pGHT1.7k and pGHT2.8k, which were used for the knockout/knockin of the endogenous GH gene of zebrafish, and injected these two vectors into the embryos of zebrafish. Overall, the rate of targeted integration with the characteristic of germ line transmission in zebrafish was 1.7x10(-6). In one experimental patch, the integrating efficiency of pGHT2.8k was higher than that of pGHT1.7k, but the lethal effect of pGHT2.8k was stronger than that of pGHT1.7k. The clones with the correct integration of target genes were identified by a simple screening procedure based on green fluorescent protein (GFP) and RFP dual selection, which corresponded to homologous recombination and random insertion, respectively. The potential homologous recombination zebrafish was further bred to produce a heterozygous F-1 generation, selected based on the presence of GFP. The potential targeted integration of exogenous GH genes into a zebrafish genome at the P-0 generation was further verified by polymerase chain reaction and Southern blot analysis. Approximately 2.5% of potential founder knockout and knockin zebrafish had the characteristic of germ line transmission. In this study, we developed an efficient method for producing the targeted gene modification in zebrafish for future studies on genetic modifications and gene functions using this model organism.
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To study the immunologic function of bursin, we analyzed the effects of anti-bursin monoclonal antibody (mAb) on the immunosuppression in ducks (Cherry Valley duck) by injecting various doses of the anti-bursin mAb into 13-d duck embryos. After hatch, cell-mediated immune activity and humoral responses were studied using lymphocyte proliferation test, tube agglutination test, and indirect enzyme-linked immuno-sorbent assay to detect anti-Escherichia coli antibodies and antibodies to Riemerella anatipester, respectively. Simultaneously, relative weights (BW-adjusted) of bursa of Fabricius (BF), spleen, and thymus were determined. Additionally, the morphology of BF, spleen, and thymus was examined at various ages using conventional histology. Follicle morphology of BF was analyzed by image analysis. The results indicated that anti-bursin mAb markedly decreased duck lymphocyte proliferation, the antibody-producing ability to bacteria, as well as the relative BF weight. Moreover, the anti-bursin mAb hindered the development of BF follicles.
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The signals of dolphins and porpoises often exhibit a multi-pulse structure. Here, echolocation signal recordings were made from four geometrically distinct positions of seven Yangtze finless porpoises temporarily housed in a relatively small, enclosed area. Some clicks demonstrated double-pulse, and others multi-pulse, structure. The interpulse intervals between the first and second pulse of the double- and multi-pulse clicks were significantly different among data from the four different positions (p < 0.01, one-way ANOVA). These results indicate that the interpulse interval and structure of the double- and multi-pulse echolocation signals depend on the hydrophone geometry of the animal, and that the double- and multi-pulse structure of echolocation signals in Yangtze finless porpoise is not caused by the phonating porpoise itself, but by the multipath propagation of the signal. Time delays in the 180 degrees phase-shifted surface reflection pulse and the nonphase-shifted bottom reflection pulse of the multi-pulse structures, relative to the direct signal, can be used to calculate the distance to a phonating animal. (c) 2005 Acoustical Society of America.
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The polarization dependence of the double resonant Raman scattering (2D) band in bilayer graphene (BLG) is studied as a function of the excitation laser energy. It has been known that the complex shape of the 2D band of BLG can be decomposed into four Lorentzian peaks with different Raman frequency shifts attributable to four individual scattering paths in the energy-momentum space. From our polarization dependence study, however, we reveal that each of the four different peaks is actually doubly degenerate in its scattering channels, i.e., two different scattering paths with similar Raman frequency shifts for each peak. We find theoretically that one of these two paths, ignored for a long time, has a small contribution to their scattering intensities but are critical in understanding their polarization dependences. Because of this, the maximum-to-minimum intensity ratios of the four peaks show a strong dependence on the excitation energy, unlike the case of single-layer graphene (SLG). Our findings thus reveal another interesting aspect of electron-phonon interactions in graphitic systems. © 2014 Elsevier Ltd. All rights reserved.
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The humoral immune responses of grouper Epinephelus akaara to a natural infection with Glugea epinephelusis was studied by ELISA utilizing intact mature spores as the coated antigen. Results showed that a specific humoral immune response was elicited, but the intensity of infection (in terms of the number of cysts) was not related to the antibody level in naturally infected hosts. The differences in the antigenicity of intact mature spores and soluble spore proteins derived from cracked mature spores were also analyzed. Results suggested that similar antigen epitopes existed between the 2 groups. Additionally, antigen component patterns and the distribution of antigen with immunogenicity were investigated by using the western blot and the immunofluorescent antibody technique (IFAT). The new parasitic microsporidium has specific polypeptide patterns comparable to the reported fish microsporidians. The main antigenic substances are concentrated on the surface of spores, and are mostly located on the anterior and posterior end of the spore bodies. Most surface components of the G. epinephelusis spores are soluble, The potential role of the surface components in initiating infection was also discussed.
Resumo:
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.
Resumo:
An antibody phage display library against White Spot Syndrome Virus (WSSV) was constructed. After four rounds of panning against WSSV, 192 out of 480 clones displayed WSSV binding activity. One of the positive clones, designated A1, had relatively higher activity specifically binding to WSSV A1-soluble, single-chain fragment variable (scFv) antibody has an affinity constant (K-aff) of 2.02 +/- 0.42 x 10(9) M-1. Dot blot assays showed that A1-soluble scFv could detect WSSV directly from shrimp hemolymph after 24-h feeding infection by WSSV. A1 scFv has potential for the development of a cheap, simple and sensitive diagnostic kit for WSSV in the field. (C) 2003 Elsevier Science B.V. All rights reserved.
Resumo:
The humoral antibody response and the number of pronephric antibody-secreting cells were examined in naturally Bothriocephalus acheilognathi-infected carp. Cyprinus carpio, and in those injected intraperitoneally with an extract of the cestode. In the extract-injected fish, specific antibody was detected 3 weeks after a second injection given 2 weeks after the primary injection, and antibody levels persisted for more than 200 days. A third injection also enhanced the antibody level in the extract-injected carp. The numbers of antibody-secreting cells were significantly higher in carp injected 3 times with the extract than in the control. In naturally-infected fish, the serum antibody levels and the number of pronephric antibody-secreting cells were higher in infected fish than in uninfected individuals although this difference was not statistically significant. The relevance of these results to immune protection against infection is discussed.