Identification and characterization of a late gene of grouper iridovirus 61l and antibody production against the protein encoded by it.

In this study, a late gene encoded by grouper iridovirus, giv-61L, was identified and classified, and mouse monoclonal antibodies (mAbs) were raised against this protein. Giv-61L homologues were found only in the genus Ranavirus. Three mAbs to Giv-61L protein were produced. In drug inhibition assays, giv-61L was identified as a late gene. Finally, GIV-61L-mAb-8 was used in western blotting and immunofluorescence assays to demonstrate that Giv-61L protein was included in the GIV particle, expressed at 18 h, and localized only in the cytoplasm of GIV-infected cells. The results of this study provide insight into GIV pathogenesis and GIV-61L-mAbs will have broad applications in GIV immunodiagnostics.

Holographic three-dimensional telepresence using large-area photorefractive polymer.

Holography is a technique that is used to display objects or scenes in three dimensions. Such three-dimensional (3D) images, or holograms, can be seen with the unassisted eye and are very similar to how humans see the actual environment surrounding them. The concept of 3D telepresence, a real-time dynamic hologram depicting a scene occurring in a different location, has attracted considerable public interest since it was depicted in the original Star Wars film in 1977. However, the lack of sufficient computational power to produce realistic computer-generated holograms and the absence of large-area and dynamically updatable holographic recording media have prevented realization of the concept. Here we use a holographic stereographic technique and a photorefractive polymer material as the recording medium to demonstrate a holographic display that can refresh images every two seconds. A 50 Hz nanosecond pulsed laser is used to write the holographic pixels. Multicoloured holographic 3D images are produced by using angular multiplexing, and the full parallax display employs spatial multiplexing. 3D telepresence is demonstrated by taking multiple images from one location and transmitting the information via Ethernet to another location where the hologram is printed with the quasi-real-time dynamic 3D display. Further improvements could bring applications in telemedicine, prototyping, advertising, updatable 3D maps and entertainment.

Identification of differentially expressed proteins in human malignant pleural effusions.

A high protein concentration in a pleural effusion makes it more likely to be a malignant than a transudative effusion. However, the variability in protein composition between these two forms of pleural effusion is not well understood. In order to compare their protein compositions, the proteomic profiles of 14 malignant and 13 transudative pleural effusions were studied using two-dimensional gel electrophoresis. Protein spots with differential expression were identified by matrix-assisted laser desorption/ionisation quadrupole time-of-flight mass spectrometry and liquid chromatography/tandem mass spectrometry. Targeted proteins were further examined by ELISA and Western immunoassay in all samples. Two-dimensional gel electrophoresis revealed seven spots whose expression was reduced in malignant pleural effusions. Four of the abnormal spots were identified as fibrinogen gamma-chain precursor, two as fibrinogen beta-chain precursor and one as pigment epithelium-derived factor. ELISA and Western immunoassay showed that pigment epithelium-derived factor levels were significantly lower in malignant than in transudative pleural effusions. It has been demonstrated that proteomic technologies may help in the elucidation of variable expression of proteins with particular functions. By applying these technologies, the level of pigment epithelium-derived factor, a potent anti-angiogenic factor, was found to be significantly lower in malignant than in transudative pleural effusions. This finding allows for further exploration regarding how underexpression of pigment epithelium-derived factor may relate to the pathogenesis of malignant pleural effusions.

Blunted renal responses to angiotensin II infusion in lifetime captopril-treated spontaneously hypertensive rats.

Previously, we had found that inhibition of the renin-angiotensin system in the early lifespan of spontaneously hypertensive rat could prevent the development of hypertension in this animal model. In the present study we evaluated the responses of blood pressure and renal function to intracerebroventricular administration of angiotensin II in long-term captopril-treated spontaneously hypertensive rats. Spontaneously hypertensive rats had been mated and their pups were treated with captopril through drinking water after birth. Age-matched Wistar-Kyoto and spontaneously hypertensive rats drinking tap water were used as control groups. At 4 months of age, the basal mean arterial blood pressure of captopril-treated hypertensive rats was the lowest among those of controlled hypertensive and normotensive rats (98+/-5 vs. 160+/-4 and 126+/-4 mmHg, respectively). Intravenous administration of angiotensin II caused similar increments of blood pressure in all rat groups. However, intracerebroventricular administration of angiotensin II to captopril-treated hypertensive rats induced a significantly less increase of arterial blood pressure in comparison with other groups. The sensitivity of baroreflex in captopril-treated hypertensive rats was also the lowest among all rat groups. The basal urine flow, sodium and potassium excretion rates, and osmolar clearance of captopril-treated hypertensive rats were significantly higher than those of controlled hypertensive rats. Intracerebroventricular infusion of angiotensin II caused significant increases in urine flow, electrolytes excretion, osmolar clearance, and free water reabsorption rate of both normotensive and controlled hypertensive rats. However, the same angiotensin II treatment did not change any of the renal excretion indices in captopril-treated hypertensive rats. Our results suggest that lifetime captopril treatment can decrease the activity of the renin-angiotensin system in the brain of hypertensive animals, which caused increases in basal urine flow and excretion of electrolytes and enhanced the sensitivity of baroreflex. It is likely that changes in the renal and baroreflex functions underlie the prevention of hypertension elicited by long-term captopril treatment.

Prenatal exposure to morphine alters kinetic properties of NMDA receptor-mediated synaptic currents in the hippocampus of rat offspring.

Whole-cell patch-clamp recordings of pharmacologically isolated N-methyl-D-asparate (NMDA) receptor-mediated evoked excitatory postsynaptic currents (EPSCs) were made, to study whether prenatal exposure to morphine affected functional properties of synaptic NMDA receptors in hippocampal slices of 2-week-old rat offspring from morphine-addicted mothers. The saturated amplitude of synaptic NMDA receptor-mediated EPSCs from morphine-treated offspring was about twofold larger than that from vehicle-control offspring. The apparent dissociation constant (Kd) values of NMDA receptors for Mg2+ at 0 mV were 7.5 +/- 1.4 and 7.9 +/- 1.3 mM in slices from vehicle-control and morphine-treated offspring, respectively. In addition, no distinguishable changes in the voltage-dependent nature and the reversal potential of NMDA receptors occurred in morphine-treated offspring, suggesting no alterations of Mg2+ blockade and ion selectivity to NMDA receptors. The 10-90% rise times of NMDA receptor-mediated EPSCs in morphine-treated offspring became longer than those in vehicle-control offspring. The decay of NMDA receptor-mediated EPSCs in both morphine-treated and vehicle-control offspring could be described by the sum of a fast and a slow exponential function. The slow, but not fast, decay times of synaptic NMDA receptor-mediated currents in morphine-treated offspring became slower than those in vehicle-control offspring. Collectively, these results suggest that prenatal exposure to morphine altered kinetic properties of synaptic NMDA receptors in hippocampal CA1 pyramidal neurons of rat offspring during early life. The extended duration of synaptic NMDA receptor-mediated currents presumably provided more Ca2+ entry through NMDA receptors in morphine-treated offspring, and its further prolongation by depolarization in such young offspring strengthened NMDA receptor-dependent functions. Thus, in light of pathophysiological implications within the central nervous system of morphine-treated offspring during early life, the present study may provide important insights and serve as a basis for therapeutic intervention in conditions under which NMDA receptors become abnormal.

Increased progenitor cell proliferation in the peripheral blood of patients with bronchial asthma: the role of nitric oxide.

BACKGROUND: Asthma exacerbation is associated with increased numbers of circulating CD34(+) progenitor cells, which may migrate to airways and develop into mature cells under the effects of cytokines and hematopoietic factors. Nitric oxide (NO) generation is enhanced in asthma and is known to suppress human hematopoiesis. OBJECTIVES: We studied circulating progenitor cells in the blood of patients with varying severity of asthma and examined the contribution of NO to their proliferation into eosinophil-forming colonies ex vivo. METHODS: With use of multiparameter flow cytometric analyses, the cell numbers and intracellular inducible NO synthase (iNOS) immunoreactivity of circulating CD34(+) cells in peripheral blood was measured. The serum level of GM-CSF or IL-5 was also determined. The colonies grown from progenitor cells were cultured in methylcellulose either in the presence or absence of growth factors, including GM-CSF, stem cell factor, and IL-3. RESULTS: A significantly greater number of circulating CD34(+) cells increased together with higher intracellular iNOS immunoreactivity in moderate asthmatics compared with mild intermittent asthmatics and healthy subjects. There was no significant difference in iNOS immunoreactivities or CD34(+) progenitor cell numbers between healthy subjects and those with mild intermittent asthma. Serum levels of GM-CSF or IL-5 were significantly higher in all asthmatics compared with healthy subjects and correlated with circulating CD34(+) cells. A greater number of colonies was grown either in the presence or absence of growth factors with a higher percentage of cells of eosinophil lineage in asthmatics than in health subjects. N(G)-nitro-L-arginine methyl ester potentiated and sodium nitroprusside inhibited the colony growth in both asthmatic and healthy subjects without a significant change in the percentage of eosinophil lineage. CONCLUSIONS: The production of NO from progenitor cells or other circulating cells may act in an autocrine or paracrine fashion to regulate progenitor cell growth and colony formation. However, this is not sufficient to control the increased proliferation of progenitor cells observed in asthma.

Activation of gonadotropin-releasing hormone receptors induces a long-term enhancement of excitatory postsynaptic currents mediated by ionotropic glutamate receptors in the rat hippocampus.

Whole-cell patch-clamp recordings were made from CA1 pyramidal neurons of the rat hippocampus to study the modulation of gonadotropin-releasing hormone (GnRH) on synaptic transmission mediated by ionotropic glutamate receptors. Leuprolide (10(-9)-10(-7) M), a specific GnRH analog, concentration-dependently elicited a long-lasting potentiation of excitatory postsynaptic currents (EPSCs) mediated by ionotropic glutamate receptors. GnRH receptor-induced synaptic potentiation was blocked by 1 microM [Acetyl-3,4-dehydro-Pro1,D-p-F-Phe2,D-Trp3,6]-LHRH, a specific GnRH receptor antagonist. Furthermore, GnRH receptor-induced synaptic potentiation was associated with the stimulation of protein kinase C (PKC), being considerably attenuated by a potent PKC inhibitor (30 microM H-7). The results suggest a long-term enhanced modulation of GnRH on synaptic transmission mediated by ionotropic glutamate receptors, possibly via the actions of PKC in the hippocampus that is an important integrative system in the regulation of reproductive processes.

High resolution electron microscopy of amorphous interlayers between metal thin films and silicon.

High-resolution electron microscopy of amorphous interlayers (a-interlayer) formed by solid-state diffusion between metal thin films and silicon is reviewed. In this paper, an overview of the development is presented. Pertinent data obtained on the growth kinetics and structure of a-interlayers in polycrystalline metal thin films on single-crystal silicon are reported. For the Ti/Si, Zr/Si, Hf/Si, V/Si, Nb/Si and Ta/Si systems, the growth of a-interlayer was found to follow a linear law in the initial stage. Si atoms were found to be the dominant diffusing species in the solid phase amorphization in the Ti/Si, Zr/Si, and Hf/Si systems. For the Y/Si system, the stability of amorphous interlayer depends critically on the composition of the amorphous films. Auto-correlation function analysis was utilized to determine the structure of the amorphous interlayers. HRTEM in conjunction with the fast Fourier transform were applied to determine the first nucleated crystalline phase. Simultaneous presence of multiphases was observed to occur in a number of refractory metal/Si systems.

The involvement of nitric oxide in synergistic neuronal damage induced by beta-amyloid peptide and glutamate in primary rat cortical neurons.

Abnormal extracellular accumulation of beta-amyloid (Abeta), the major component of senile plaques in the Alzheimer's brain and the excitatory amino acid glutamate are both considered to be associated with neurodegeneration. We studied whether nitric oxide (NO) was involved in neuronal damage induced by Abeta and glutamate in primary rat cortical neurons. Our results demonstrated that (1) Both neuronal damage and NO production were synergistically induced by Abeta-(25-35) and glutamate; (2) This synergistic neuronal damage induced by Abeta-(25-35) and glutamate was attenuated by selective inhibitors of NO synthase. We propose that cytotoxic characteristics of NO, at least in part, are involved in the synergistic neuronal damage induced by Abeta and glutamate, presumably seen in Alzheimer's brains.

Free radicals are involved in methylmethacrylate-induced neurotoxicity in human primary neocortical cell cultures.

Methylmethacrylate monomer (MMA), a highly volatile material, has been extensively used for the construction of complete or partial dental prostheses. While previous studies have indicated a variety of complications and untoward side-effects associated with its use, the possible neurotoxicity induced by this monomer has not been addressed. In this study, we have investigated the MMA-produced neuronal injury in human neuron-enriched primary culture. Embryonic brain tissue (8-10 weeks postconception) was used for the primary neuron-enriched culture. Phase-contrast microscopy was used to evaluate morphological changes of cultured neurons. Extracellular concentrations of lactate dehydrogenase (LDH) and nitrite was measured from the culture medium to assess the magnitude of neuronal damage and nitric oxide formation, respectively. Neocortical neurons exposed to the monomer (1/200, Vmonomer/Vglycerol) for two days resulted in a significant increase in the LDH level but monomer (1/20000, 1/2000, or 1/200; Vmonomer/Vglycerol) failed to increase the nitrite level. Morphologically, the neurons subjected to monomer treatment exhibited irregular shrunken cell bodies with dystrophic and/or fragmented neurities, or even cell lysis. Moreover, superoxide dismutase plus catalase or vitamin C pretreatment protected against monomer-induced neurotoxicity. Our results suggest that this neurotoxicity can not likely be attributed to the cytotoxic effects of nitric oxide but may be mediated through the toxicity of superoxide and other free radicals. This is the first time, to our knowledge, that neurotoxicity induced by MMA has been demonstrated in human cortical neurons.