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.

Estradiol stimulates growth hormone production in female goldfish

The effects of estradiol (E(2)) on growth hormone (GH) production was investigated in gonad-intact female goldfish. It was first necessary to generate a specific antibody for use in immunocytochemistry, Western, and dot-blot analyses of GH production. To accomplish this, grass carp GH (gcGH) cDNA was cloned by the reverse transcription polymerase chain reaction (RT-PCR) and expressed in Echerichia coli and a specific polyclonal antibody to recombinant gcGH was generated in the rabbit. In Western blot, the anti-gcGH antibody specifically immunoreacted with recombinant gcGH, purified natural common carp GH, and with a single 21.5-kDa GH form from pituitary extracts of grass carp, common carp, goldfish, and zebrafish but not salmon, trout, or tilapia. Intraperitoneal injection of the recombinant gcGH enhanced the growth rates of juvenile common carp demonstrating biological activity of this GH preparation. Electron microscopic studies showed that the anti-gcGH-I antibody specifically reacted with GH localized in the secretory granules of the goldfish somatotroph. Using anti-gcGH-I in a dot-blot assay, it was found that in vivo implantation of solid silastic pellets containing E(2) (100 mu g/g body weight for 5 days) increased pituitary GH content by 150% in female goldfish. In a second, independent study employing a previously characterized anticommon carp GH antibody for radioimmunoassay, it was found that E(2) increased pituitary GH content by 170% and serum GH levels by approximately 350%. The E(2)-induced hypersecretion of GH and increase in pituitary GH levels was not associated with changes in steady-state pituitary GH mRNA levels, suggesting that this sex steroid may enhance GH synthesis at the posttranscriptional or translational level. Previous observations indicate that GH can stimulate ovarian E(2) production. The present results show that E(2) can in turn stimulate GH production, indicating the existence of a novel pituitary GH-ovarian feedback system in goldfish. (C) 1997 Academic Press.

Stiction and Anti-Stiction in Mems and Nems

Stiction in microelectromechanical systems (MEMS) has been a major failure mode ever since the advent of surface micromachining in the 80s of the last century due to large surface-area-to-volume ratio. Even now when solutions to this problem are emerging, such as self-assembled monolayer (SAM) and other measures, stiction remains one of the most catastrophic failure modes in MEMS. A review is presented in this paper on stiction and anti-stiction in MEMS and nanoelectromechanical systems (NEMS). First, some new experimental observations of stiction in radio frequency (RF) MEMS switch and micromachined accelerometers are presented. Second, some criteria for stiction of microstructures in MEMS and NEMS due to surface forces (such as capillary, electrostatic, van der Waals, Casimir forces, etc.) are reviewed. The influence of surface roughness and environmental conditions (relative humidity and temperature) on stiction are also discussed. As hydrophobic films, the self-assembled monolayers (SAMs) turn out able to prevent release-related stiction effectively. The anti-stiction of SAMs in MEMS is reviewed in the last part.

Stress intensity factors calculation in anti-plane fracture problem by orthogonal integral extraction method based on femol

For an anti-plane problem, the differential operator is self-adjoint and the corresponding eigenfunctions belong to the Hilbert space. The orthogonal property between eigenfunctions (or between the derivatives of eigenfunctions) of anti-plane problem is exploited. We developed for the first time two sets of radius-independent orthogonal integrals for extraction of stress intensity factors (SIFs), so any order SIF can be extracted based on a certain known solution of displacement (an analytic result or a numerical result). Many numerical examples based on the finite element method of lines (FEMOL) show that the present method is very powerful and efficient.

Unloading characteristics of anti-plane shear crack in softening materials

The local characteristics of the anti-plane shear stress and strain field are determined for a material where the stress increases linearly with strain up to a limit and then softens nonlinearly. Two unloading models are considered such that the unloading path always returns to the origin while the other assumes the unloading modulus to be that of the initial shear modulus. As the applied shear increases, an unloading zone is found to prevail between a zone in which the material softens and another zone in which the material is linear-elastic even though the crack does not propagate. The divisions of these zones are displayed graphically.

The Mitogenic and Anti-Apoptotic Activity of Tumor Conditioned Medium on Endothelium

This study was designed to observe the effect of tumor conditioned medium (TCM) on the proliferation and apoptosis of human umbilical vein endothelial cells (HUVECs). HUVECs were exposed to TCM from breast carcinoma cell line MDA-MB-231, then we measured their proliferation, apoptosis and cell cycle distribution by MTT and flow cytometery (FCM). Following the stimulation of TCM, HUVECs showed higher pro-mitogenic and anti-apoptotic ability than did the negative control group (ECGF-free medium with 20% FBS), but a similar ability to the positive control group (medium with ECGF and 20% FBS). From these results, we can conclude that breast carcinoma cell line MDA-MB-231 could secret soluble pro-angiogenic factors that induce HUVEC angiogenic switching, including cell cycle progression, proliferation and growth. The role and character of these factors remain to be further studied.

Imaging properties of coherent anti-Stokes Raman scattering microscope

The coherent anti-Stokes Raman scattering (CARS) microscope with the combination of confocal and CARS techniques is a remarkable alternative for imaging chemical or biological specimens that neither fluoresce nor tolerate labelling. CARS is a nonlinear optical process, the imaging properties of CARS microscopy will be very different from the conventional confocal microscope. In this paper, the intensity distribution and the polarization property of the optical field near the focus was calculated. By using the Green function, the precise analytic solution to the wave equation of a Hertzian dipole source was obtained. We found that the intensity distributions vary considerably with the different experimental configurations and the different specimen shapes. So the conventional description of microscope (e.g. the point spread function) will fail to describe the imaging properties of the CARS microscope.

The intensity distributions of collected signals in coherent anti-Stokes Raman scattering microscopy

Coherent anti-Stokes Raman scattering (CARS) microscopy with the combining of confocal and CARS techniques is a remarkable alternative for imaging chemical or biological specimens that neither fluoresce nor tolerate labeling. The CARS is a nonlinear optical process, the imaging properties of CARS microscopy will be very different from the conventional confocal microscopy. In this paper, we calculated the propagation of CARS signals by using the wave equation in medium and the slowly varying envelope approximation (SVEA), and find that the intensity angular distributions vary considerably with the different experimental configurations and the different specimen shapes. So the conventional description of microscopy (e.g.. the point spread function) will fail to descript the imaging properties of CARS microscopy. (c) 2004 Elsevier B.V. All rights reserved.

Encoded cell grating array in anti-counterfeit technology

The dot matrix hologram (DMH) has been widely used in anti-counterfeiting label. With the same technology and cell array configuration, we can encode to the incidence beam. These codes can be some image matrix grating with different grating gap and different grating orientation. When the multi-level phase diffractive grating is etched, the incidence beam on the cell appears as an encoding image. When the encoded grating and DMH are used in the same label synchronously, the technology of multi-encoded grating array enhances the anti-counterfeit ability.

Influence of vacuum organic contaminations on laser-induced damage of 1064 nm anti-reflective coatings

We investigate the influence of vacuum organic contaminations on laser-induced damage threshold (LIDT) of optical coatings. Anti-reflective (AR) coatings at 1064 nm made by Ta2O5/SiO2 are deposited by the ion beam sputtering method. The LIDTs of AR coatings are measured in vacuum and in atmosphere, respectively. It is exhibited that contaminations in vacuum are easily to be absorbed onto optical surface because of lower pressure, and they become origins of damage, resulting in the decrease of LIDT from 24.5 J/cm(2) in air to 15.7 J/cm(2) in vacuum. The LIDT of coatings in vacuum has is slightly changed compared with the value in atmosphere after the organic contaminations are wiped off. These results indicate that organic contaminations are the main reason of the LIDT decrease in vacuum. Additionally, damage morphologies have distinct changes from vacuum to atmosphere because of the differences between the residual stress and thermal decomposability of filmy materials.