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Selected AbstractsTGF-,3,dependent SMAD2 phosphorylation and inhibition of MEE proliferation during palatal fusionDEVELOPMENTAL DYNAMICS, Issue 3 2003Xiao-Mei Cui Abstract Transforming growth factor (TGF) -,3 is known to selectively regulate the disappearance of murine medial edge epithelium (MEE) during palatal fusion. Previous studies suggested that the selective function of TGF-,3 in MEE was conducted by TGF-, receptors. Further studies were needed to demonstrate that the TGF-, signaling mediators were indeed expressed and phosphorylated in the MEE cells. SMAD2 and SMAD3 were both present in the MEE, whereas SMAD2 was the only one phosphorylated during palatal fusion. SMAD2 phosphorylation was temporospatially restricted to the MEE and correlated with the disappearance of the MEE. No phosphorylated SMAD2 was found in MEE in TGF-,3,/, mice, although nonphosphorylated SMAD2 was present. The results suggest that TGF-,3 is required for initiating and maintaining SMAD2 phosphorylation in MEE. Phospho-SMAD3 was not detectable in palate during normal palatal fusion. Previous results suggested TGF-,,induced cessation of DNA synthesis in MEE cells during palatal fusion in vitro. The present results provide evidence that inhibition of MEE proliferation in vivo was controlled by endogenous TGF-,3. The number of 5-bromo-2,-deoxyuridine (BrdU) -labeled MEE cells was significantly reduced in TGF-,3+/+ compared with TGF-,3,/, mice when the MEE seam formed (t -test, P < 0.05). This finding suggests that TGF-,3 is required for inhibiting MEE proliferation during palatal fusion. The inhibition of MEE proliferation may be mediated by TGF-,3,dependent phosphorylation of SMAD2. Developmental Dynamics 227:387,394, 2003. © 2003 Wiley-Liss, Inc. [source] WITHIN-HOST POPULATION DYNAMICS AND THE EVOLUTION OF MICROPARASITES IN A HETEROGENEOUS HOST POPULATIONEVOLUTION, Issue 2 2002Vitaly V. Ganusov Abstract Why do parasites harm their hosts? The general understanding is that if the transmission rate and virulence of a parasite are linked, then the parasite must harm its host to maximize its transmission. The exact nature of such trade-offs remains largely unclear, but for vertebrate hosts it probably involves interactions between a microparasite and the host immune system. Previous results have suggested that in a homogeneous host population in the absence of super- or coinfection, within-host dynamics lead to selection of the parasite with an intermediate growth rate that is just being controlled by the immune system before it kills the host (Antia et al. 1994). In this paper, we examine how this result changes when heterogeneity is introduced to the host population. We incorporate the simplest form of heterogeneity,random heterogeneity in the parameters describing the size of the initial parasite inoculum, the immune response of the host, and the lethal density at which the parasite kills the host. We find that the general conclusion of the previous model holds: parasites evolve some intermediate growth rate. However, in contrast with the generally accepted view, we find that virulence (measured by the case mortality or the rate of parasite-induced host mortality) increases with heterogeneity. Finally, we link the within-host and between-host dynamics of parasites. We show how the parameters for epidemiological spread of the disease can be estimated from the within-host dynamics, and in doing so examine the way in which trade-offs between these epidemiological parameters arise as a consequence of the interaction of the parasite and the immune response of the host. [source] Human airway trypsin-like protease induces amphiregulin release through a mechanism involving protease-activated receptor-2-mediated ERK activation and TNF ,-converting enzyme activity in airway epithelial cellsFEBS JOURNAL, Issue 24 2005Manabu Chokki Human airway trypsin-like protease (HAT), a serine protease found in the sputum of patients with chronic airway diseases, is an agonist of protease-activated receptor-2 (PAR-2). Previous results have shown that HAT enhances the release of amphiregulin (AR); further, it causes MUC5AC gene expression through the AR-epidermal growth factor receptor pathway in the airway epithelial cell line NCI-H292. In this study, the mechanisms by which HAT-induced AR release can occur were investigated. HAT-induced AR gene expression was mediated by extracellular signal-regulated kinase (ERK) pathway, as pretreatment of cells with ERK pathway inhibitor eliminated the effect of HAT on AR mRNA. Both HAT and PAR-2 agonist peptide (PAR-2 AP) induced ERK phosphorylation; further, desensitization of PAR-2 with a brief exposure of cells to PAR-2 AP resulted in inhibition of HAT-induced ERK phosphorylation, suggesting that HAT activates ERK through PAR-2. Moreover, PAR-2 AP induced AR gene expression subsequent to protein production in the cellular fraction through the ERK pathway indicating that PAR-2-mediated activation of ERK is essential for HAT-induced AR production. However, in contrast to HAT, PAR-2 AP could not cause AR release into extracellular space; it appears that activation of PAR-2 is not sufficient for HAT-induced AR release. Finally, HAT-induced AR release was eliminated by blockade of tumour necrosis factor ,-converting enzyme (TACE) by the TAPI-1 and RNA interference, suggesting that TACE activity is necessary for HAT-induced AR release. These observations show that HAT induces AR production through the PAR-2 mediated ERK pathway, and then causes AR release by a TACE-dependent mechanism. [source] ,-Fetoprotein positively regulates cytochrome c -mediated caspase activation and apoptosome complex formationFEBS JOURNAL, Issue 21 2003Lidia Semenkova Previous results have shown that the oncoembryonic marker ,-fetoprotein (AFP) is able to induce apoptosis in tumor cells through activation of caspase 3, bypassing Fas-dependent and tumor necrosis factor receptor-dependent signaling. In this study we further investigate the molecular interactions involved in the AFP-mediated signaling of apoptosis. We show that AFP treatment of tumor cells is accompanied by cytosolic translocation of mitochondrial cytochrome c. In a cell-free system, AFP mediates processing and activation of caspases 3 and 9 by synergistic enhancement of the low-dose cytochrome c -mediated signals. AFP was unable to regulate activity of caspase 3 in cell extracts depleted of cytochrome c or caspase 9. Using high-resolution chromatography, we show that AFP positively regulates cytochrome c/dATP-mediated apoptosome complex formation, enhances recruitment of caspases and Apaf-1 into the complex, and stimulates release of the active caspases 3 and 9 from the apoptosome. By using a direct protein,protein interaction assay, we show that pure human AFP almost completely disrupts the association between processed caspases 3 and 9 and the cellular inhibitor of apoptosis protein (cIAP-2), demonstrating its release from the complex. Our data suggest that AFP may regulate cell death by displacing cIAP-2 from the apoptosome, resulting in promotion of caspase 3 activation and its release from the complex. [source] The N -acetylglutamate synthase/N -acetylglutamate kinase metabolon of Saccharomyces cerevisiae allows co-ordinated feedback regulation of the first two steps in arginine biosynthesisFEBS JOURNAL, Issue 5 2003Katia Pauwels In Saccharomyces cerevisiae, which uses the nonlinear pathway of arginine biosynthesis, the first two enzymes, N -acetylglutamate synthase (NAGS) and N -acetylglutamate kinase (NAGK), are controlled by feedback inhibition. We have previously shown that NAGS and NAGK associate in a complex, essential to synthase activity and protein level [Abadjieva, A., Pauwels, K., Hilven, P. & Crabeel, M. (2001) J. Biol. Chem.276, 42869,42880]. The NAGKs of ascomycetes possess, in addition to the catalytic domain that is shared by all other NAGKs and whose structure has been determined, a C-terminal domain of unknown function and structure. Exploring the role of these two domains in the synthase/kinase interaction, we demonstrate that the ascomycete-specific domain is required to maintain synthase activity and protein level. Previous results had suggested a participation of the third enzyme of the pathway, N -acetylglutamylphosphate reductase, in the metabolon. Here, genetic analyses conducted in yeast at physiological level, or in a heterologous background, clearly demonstrate that the reductase is dispensable for synthase activity and protein level. Most importantly, we show that the arginine feedback regulation of the NAGS and NAGK enzymes is mutually interdependent. First, the kinase becomes less sensitive to arginine feedback inhibition in the absence of the synthase. Second, and as in Neurospora crassa, in a yeast kinase mutant resistant to arginine feedback inhibition, the synthase becomes feedback resistant concomitantly. We conclude that the NAGS/NAGK metabolon promotes the co-ordination of the catalytic activities and feedback regulation of the first two, flux controlling, enzymes of the arginine pathway. [source] A folding variant of human ,-lactalbumin induces mitochondrial permeability transition in isolated mitochondriaFEBS JOURNAL, Issue 1 2001Camilla Köhler A human milk fraction containing multimeric ,-lactalbumin (MAL) is able to kill cells via apoptosis. MAL is a protein complex of a folding variant of ,-lactalbumin and lipids. Previous results have shown that upon treatment of transformed cells, MAL localizes to the mitochondria and cytochrome c is released into the cytosol. This is followed by activation of the caspase cascade. In this study, we further investigated the involvement of mitochondria in apoptosis induced by the folding variant of ,-lactalbumin. Addition of MAL to isolated rat liver mitochondria induced a loss of the mitochondrial membrane potential (,,m), mitochondrial swelling and the release of cytochrome c. These changes were Ca2+ -dependent and were prevented by cyclosporin A, an inhibitor of mitochondrial permeability transition. MAL also increased the rate of state 4 respiration in isolated mitochondria by exerting an uncoupling effect. This effect was due to the presence of fatty acids in the MAL complex because it was abolished completely by BSA. BSA delayed, but failed to prevent, mitochondrial swelling as well as dissipation of ,,m, indicating that the fatty acid content of MAL facilitated, rather than caused, these effects. Similar results were obtained with HAMLET (human ,-lactalbumin made lethal to tumour cells), which is native ,-lactalbumin converted in vitro to the apoptosis-inducing folding variant of the protein in complex with oleic acid. Our findings demonstrate that a folding variant of ,-lactalbumin induces mitochondrial permeability transition with subsequent cytochrome c release, which in transformed cells may lead to activation of the caspase cascade and apoptotic death. [source] The effect of superoxide dismutase deficiency on cadmium stressJOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY, Issue 1 2004Paula D. B. Adamis Abstract Saccharomyces cerevisiae mutant strains deficient in superoxide dismutase (Sod), an antioxidant enzyme, were used to analyze cadmium absorption and the oxidation produced by it. Cells lacking the cytosolic Sod1 removed twice as much cadmium as the control strain, while those deficient in the mitochondrial Sod2 exhibited poor metal absorption. Interestingly, the sod1 mutant did not become more oxidized after exposure to cadmium, as opposed to the control strain. We observed that the deficiency of Sod1 increases the expression of both Cup1 (a metallothionein) and Ycf1 (a vacuolar glutathione S-conjugate pump), proteins involved with protection against cadmium. Furthermore, when sod1 cells were exposed to cadmium, the ratio glutathione oxidized/glutathione reduced did not increase as expected. We propose that a high level of metallothionein expression would relieve glutathione under cadmium stress, while an increased level of Ycf1 expression would favor compartmentalization of this metal into the vacuole. Both conditions would reduce the level of glutathione-cadmium complex in cytosol, contributing to the high capacity of absorbing cadmium by the sod1 strain. Previous results showed that the glutathione-cadmium complex regulates cadmium uptake. These results indicate that, even indirectly, metallothionein also regulates cadmium transport. © 2004 Wiley Periodicals, Inc. J Biochem Mol Toxicol 18:12,17, 2004; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.20000 [source] Molecular characterization of protein kinase C-, binding to lamin AJOURNAL OF CELLULAR BIOCHEMISTRY, Issue 2 2002Alberto M. Martelli Abstract Previous results from our laboratory have identified lamin A as a protein kinase C (PKC)-binding protein. Here, we have identified the regions of PKC-, that are crucial for this binding. By means of overlay assays and fusion proteins made of glutathione-S-transferase (GST) fused to elements of rat PKC-,, we have established that binding occurs through both the V5 region and a portion of the C2 region (i.e., the calcium-dependent lipid binding (CaLB) domain) of the kinase. In particular, we have found that amino acid 200,217 of the CaLB domain are essential for binding lamin A, as a synthetic peptide corresponding to this stretch of amino acids prevented the interaction between the CaLB domain and lamin A. We also show that the presence of four lysine residues of the CaLB domain (K205, K209, K211, and K213) was essential for the binding. We have determined that binding of elements of PKC-, to lamin A does not require the presence of cofactors such as phosphatidylserine (PS) and Ca2+. We have also found that the binding site of lamin A for the CaLB domain of PKC-, is localized in the carboxyl-terminus of the lamin, downstream of amino acid 499. Our findings may prove to be important to clarify the mechanisms regulating PKC function within the nucleus and may also lead to the synthesis of isozyme-specific drugs to attenuate or reverse PKC-dependent nuclear signaling pathways important for the pathogenesis of cancer. © 2002 Wiley-Liss, Inc. [source] Changes in Apple Liquid Phase Concentration throughout Equilibrium in Osmotic DehydrationJOURNAL OF FOOD SCIENCE, Issue 2 2007J.M. Barat ABSTRACT:, Previous results on apple tissue equilibration during osmotic dehydration showed that, at very long processing times, the solute concentrations of the fruit liquid phase and the osmotic solution were the same. In the present study, changes in apple liquid phase composition throughout equilibrium in osmotic dehydration were analyzed and modeled. Results showed that, by the time osmosed samples reached the maximum weight and volume loss, solute concentration of the fruit liquid phase was higher than that of the osmotic solution. The reported overconcentration could be explained in terms of the apple structure shrinkage that occurred during the osmotic dehydration with highly concentrated osmotic solutions due to the elastic response of the food structure to the loss of water and intake of solutes. The fruit liquid phase overconcentration rate was observed to depend on the concentration of the osmotic solution, the processing temperature, the sample size, and shape of the cellular tissue. [source] Delayed Failure of Hi-Nicalon and Hi-Nicalon S Multifilament Tows and Single Filaments at Intermediate Temperatures (500°,800°C)JOURNAL OF THE AMERICAN CERAMIC SOCIETY, Issue 3 2009W. Gauthier Previous results have shown that tows of SiC Nicalon fibers are sensitive to the phenomenon of delayed failure, at temperatures below 700°C. The present paper examines the static fatigue of Hi-Nicalon and Hi-Nicalon S when subjected to constant load, at temperatures between 500° and 800°C in air. Multifilament tows and single filaments were tested. Experimental data show that the rupture times of tows depend on the applied stress according to the conventional power law t,n=A. In contrast, the stress-rupture time data obtained on single filaments exhibit significant scatter. A model based on slow crack growth in single filaments shows that the stress-rupture of fiber tows follows the conventional time power law. The dependence on temperature was introduced. The model allowed sound calculations of tow lifetimes and characteristics of the slow crack growth phenomenon to be extracted from the tow stress-rupture time data. [source] Impact of Chronic Anticholesterol Therapy on Development of Microvascular Rarefaction in the Metabolic SyndromeMICROCIRCULATION, Issue 8 2009Adam G. Goodwill ABSTRACT Objective: The obese Zucker rat (OZR) model of the metabolic syndrome is partly characterized by moderate hypercholesterolemia, in addition to other contributing comorbidities. Previous results suggest that vascular dysfunction in OZR is associated with chronic reduction in vascular nitric-oxide (NO) bioavailability and chronic inflammation, both frequently associated with hypercholesterolemia. As such, we evaluated the impact of chronic cholesterol-reducing therapy on the development of impaired skeletal muscle arteriolar reactivity and microvessel density in OZR and its impact on chronic inflammation and NO bioavailability. Materials and Methods: Beginning at seven weeks of age, male OZR were treated with gemfibrozil, probucol, atorvastatin, or simvastatin (in chow) for 10 weeks. Subsequently, plasma and vascular samples were collected for biochemical/molecular analyses, while arteriolar reactivity and microvessel network structure were assessed by using established methodologies after 3, 6, and 10 weeks of drug therapy. Results: All interventions were equally effective at reducing total cholesterol, although only the statins also blunted the progressive reductions to vascular NO bioavailability, evidenced by greater maintenance of acetylcholine-induced dilator responses, an attenuation of adrenergic constrictor reactivity, and an improvement in agonist-induced NO production. Comparably, while minimal improvements to arteriolar wall mechanics were identified with any of the interventions, chronic statin treatment reduced the rate of microvessel rarefaction in OZR. Associated with these improvements was a striking statin-induced reduction in inflammation in OZR, such that numerous markers of inflammation were correlated with improved microvascular reactivity and density. However, using multivariate discriminant analyses, plasma RANTES (regulated on activation, normal T-cell expressed and secreted), interleukin-10, monocyte chemoattractant protein-1, and tumor necrosis factor alpha were determined to be the strongest contributors to differences between groups, although their relative importance varied with time. Conclusions: While the positive impact of chronic statin treatment on vascular outcomes in the metabolic syndrome are independent of changes to total cholesterol, and are more strongly associated with improvements to vascular NO bioavailability and attenuated inflammation, these results provide both a spatial and temporal framework for targeted investigation into mechanistic determinants of vasculopathy in the metabolic syndrome. [source] Microvascular Display of Xanthine Oxidase and NADPH Oxidase in the Spontaneously Hypertensive RatMICROCIRCULATION, Issue 7 2006FRANK A. DELANO ABSTRACT Objective: Oxygen free radical production in hypertension may be associated with elevated arteriolar tone and organ injury. Previous results suggest an enhanced level of oxygen free radical formation in microvascular endothelium and in circulating neutrophils associated with xanthine oxidase activity in the spontaneously hypertensive rats (SHR) compared with their normotensive controls, the Wistar Kyoto rats (WKY). The aim of this study was to gain more detailed understanding of where oxidative enzymes are located in the microcirculation. Methods: An approach was developed to delineate the cellular distribution of two selected oxidative enzymes, xanthine oxidase and nicotinamide adenine dinucleotide phosphate (NADPH) dependent oxidase (protein 67-kDa fraction). Immunolabeling with peroxidase substrate was utilized, which permits full delineation of the primary antibody in all microvascular structures of the mesentery. Results: Xanthine oxidase is present in the endothelium of all segments of the microcirculation, in mast cells, and in parenchymal cells of the mesentery. NADPH oxidase can be detected in the endothelium, leukocytes, and mast cells and with lower levels in parenchymal cells. The mesentery of WKY and SHR has similar enzyme distributions with enhancements on the arteriolar and venular side of the microcirculation that coincide with the sites of enhanced free radical production recently reported. Immune label measurements under standardized conditions indicate that both enzymes are significantly enhanced in the SHR. Adrenalectomy, which serves to reduce the blood pressure and free radical production of the SHR to normotensive levels, leads to a reduction of NADPH and xanthine oxidase to normotensive levels, while supplementation of adrenalectomized SHR with dexamethasone significantly increases the oxidase expression in several parts of the microcirculation to levels above the WKY rats. Conclusion: The results indicate that enhanced expression of NADPH and xanthine oxidase in the SHR depends on an adrenal pathway that is detectable in the arteriolar and venular network at high and low pressure regions of the circulation. [source] Random dense bipartite graphs and directed graphs with specified degreesRANDOM STRUCTURES AND ALGORITHMS, Issue 2 2009Catherine Greenhill Abstract Let s and t be vectors of positive integers with the same sum. We study the uniform distribution on the space of simple bipartite graphs with degree sequence s in one part and t in the other; equivalently, binary matrices with row sums s and column sums t. In particular, we find precise formulae for the probabilities that a given bipartite graph is edge-disjoint from, a subgraph of, or an induced subgraph of a random graph in the class. We also give similar formulae for the uniform distribution on the set of simple directed graphs with out-degrees s and in-degrees t. In each case, the graphs or digraphs are required to be sufficiently dense, with the degrees varying within certain limits, and the subgraphs are required to be sufficiently sparse. Previous results were restricted to spaces of sparse graphs. Our theorems are based on an enumeration of bipartite graphs avoiding a given set of edges, proved by multidimensional complex integration. As a sample application, we determine the expected permanent of a random binary matrix with row sums s and column sums t. © 2009 Wiley Periodicals, Inc. Random Struct. Alg., 2009 [source] A study of Mg and K isotopes in Allende CAIs: Implications to the time scale for the multiple heating processesMETEORITICS & PLANETARY SCIENCE, Issue 12 2006Motoo Ito The 7R-19,1 contains 16O-rich and 16O-poor melilite grains and define a single isochron corresponding to an initial 26Al/27Al ratio of (6.6 ± 1.3) × 10,5. The Al-Mg isochron, O isotope measurements and petrography of melilite in 7R-19,1 indicate that 16O-poor melilite crystallized within 0.4 Myr after crystallization of 16O-rich melilite, suggesting that oxygen isotopic composition of the CAI-forming region changed from 16O-rich to 16O-poor within this time interval. The 16O-poor melilite is highly depleted in K compared to the adjacent 16O-rich melilite, indicating evaporation during remelting of 7R-19,1. We determined the isochron for 41Ca- 41K isotopic systematics in EGG3 pyroxene with (4.1 ± 2.0) × 10,9 (2s,) as an initial ratio of 41Ca/40Ca, which is at least two times smaller than the previous result (Sahijipal et al. 2000). The ratio of 41Ca/40Ca in the EGG3 pyroxene grain agrees within error with the value obtained by Hutcheon et al. (1984). No evidence for the presence of 41K excess (decay product of a short-lived radionuclide 41Ca) was found in 7R-19,1 and HN3,1. We infer that the CAI had at least an order of magnitude lower than canonical 41Ca/40Ca ratio at the time of the CAI formation. [source] Eosinophil cationic protein damages protoscoleces in vitro and is present in the hydatid cystPARASITE IMMUNOLOGY, Issue 8 2006A. L. RAMOS SUMMARY Eosinophils are locally recruited during the establishment and chronic phases of cystic hydatidosis. This study provides evidence that eosinophil cationic protein (ECP), one of the major components of eosinophil granules, can damage Echinococcus granulosus protoscoleces (PSC). The toxicity of ECP was investigated in vitro by following parasite viability in the presence of this protein. ECP was found to damage PSC at micromolar concentrations; the effect was blocked by specific antibodies and heparin, and was more severe than the one caused by similar concentrations of RNase A, suggesting that the cationic nature of ECP, and not its ribonuclease activity, is involved in toxicity. This observation may highlight the capacity of eosinophils to control secondary hydatidosis, derived from PSC leakage from a primary cyst. To further assess the relevance of the previous result during infection, the presence of eosinophil proteins was investigated in human hydatid cysts. ECP was found to be strongly associated with the laminated layer of the cyst wall, and present at micromolar concentrations in the hydatid fluid. Overall, these results demonstrate that eosinophils degranulate in vivo at the host,parasite interface, and that the released ECP reaches concentrations that could be harmful for the parasite. [source] An order-based algorithm for implementing multiparty synchronizationCONCURRENCY AND COMPUTATION: PRACTICE & EXPERIENCE, Issue 12 2004José A. Pérez Abstract Multiparty interactions are a powerful mechanism for coordinating several entities that need to cooperate in order to achieve a common goal. In this paper, we present an algorithm for implementing them that improves on previous results in that it does not require the whole set of entities or interactions to be known at compile- or run-time, and it can deal with both terminating and non-terminating systems. We also present a comprehensive simulation analysis that shows how sensitive to changes our algorithm is, and compare the results with well-known proposals by other authors. This study proves that our algorithm still performs comparably to other proposals in which the set of entities and interactions is known beforehand, but outperforms them in some situations that are clearly identified. In addition, these results prove that our algorithm can be combined with a technique called synchrony loosening without having an effect on efficiency. Copyright © 2004 John Wiley & Sons, Ltd. [source] Thermodynamic Properties and Plasma Phase Transition in dense HydrogenCONTRIBUTIONS TO PLASMA PHYSICS, Issue 5-6 2004V. S. Filinov Abstract The internal energy and equation of state of dense hydrogen are investigated by direct path integral Monte Carlo method simulations which are further improved in comparison to our previous results. Data for four isotherms , T = 10, 000K, 30, 000K, 50, 000K, and 100, 000K , are presented. For T = 10, 000K it is shown that the internal energy is lowered due to droplet formation for densities of the order 1023cm,3 giving direct support for the existence of a plasma phase transition in megabar hydrogen. (© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Growth of lead bromide polycrystalline filmsCRYSTAL RESEARCH AND TECHNOLOGY, Issue 10 2004M. Giles Abstract Lead bromide polycrystalline films were grown by the physical vapor deposition method (PVD). Glass 1,x1, in size, uncoated, and coated with Indium Tin Oxide (ITO), was used as substrate and rear contact. The starting material was evaporated at temperatures from 395°C to 530°C under high vacuum atmosphere (6 x 10 -3 Pa) and during 8 days. The substrate temperature was prefixed from 190°C to 220°C. Film thickness yielded values from 40 to 90 ,m. Optical microscopy and scanning electron microscopy (SEM) were performed on the films. Grain size resulted to be from 1.0 to 3.5 ,m. SEM and X-ray diffraction indicate that films grow with a preferred orientation with the (0 0 l) planes parallel to the substrate. The Texture Coefficient (TC) related to the plane (0 0 6) was 7.3. Resistivity values in the order of 1012 ,cm were obtained for the oriented samples, but a strong polarization indicates severe charge transport problems in the films. Film properties were correlated with the growth temperature and with previous results for films of other halides. (© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Discrepancy in measuring CD4 expression on T-lymphocytes using fluorescein conjugates in comparison with unimolar CD4-phycoerythrin conjugates,,CYTOMETRY, Issue 6 2007Lili Wang Abstract Background: Numerous methods for quantitative fluorescence calibration (QFC) have been developed to quantify receptor expression on lymphocytes. However, the results from the use of these different QFC methods vary considerably in the literature. To better identify the causes of these discrepancies, we measured CD4 expression using FITC and phycoerythrin (PE) conjugates to stain CYTO-TROLÔ Control Cells and T-lymphocytes in whole blood and isolated cell preparations. We further examined pH of the cellular microenvironment as a cause of discordant results obtained with the FITC conjugate. Methods: Calibration with Quantibrite PE-labeled microspheres and the use of unimolar CD4-PE conjugates provided direct measurement of the antibody bound per cell value (ABC) for CD4 expression on normal T-lymphocytes. Calibration for CD4-FITC monoclonal antibody (Mab) labeled CYTO-TROL Control Cells and normal T-lymphocytes was based on molecules of equivalent soluble fluorochrome (MESF) as determined by FITC-labeled microspheres traceable to NIST RM 8640. The MESF value for CD4-FITC Mab was determined that enabled the conversion of the MESF values obtained for CYTO-TROL cells to ABC. We investigated the likely pH change in the fluorescein microenvironments within FITC-labeled Mab and cells stained with FITC-labeled Mab using a pH sensitive indicator. Results: The mean ABC value for T-lymphocytes prepared from fresh whole blood using CD4-PE conjugate (48,321) was consistent with previous results, and it was much higher than the mean ABC using CD4-FITC Mab (22,156). The mean ABC value for CYTO-TROL cells using CD4-PE conjugate (43,090) was also higher than that using CD4-FITC conjugate (34,734), although the discrepancy was not as great. Further studies suggested the discrepancy in CYTO-TROL results may be accounted for by the low pH of the membrane microenvironment, but the greater discrepancy in T-lymphocytes could not be fully explained. Conclusion: CD4 expression on fresh normal whole blood samples and CYTO-TROL cells can be consistently quantified in ABC units using Quantibrite PE quantification beads and unimolar CD4-PE conjugates. Quantification with CD4-FITC conjugate is not as consistent, but may be improved by the use of CD4 T-cells as biological calibrators. This approximation is valid only for surface receptors with consensus ABC values measured by different QFC methods serving as biological standards. Published 2007 Wiley-Liss, Inc. [source] Structural and functional effects of hydrostatic pressure on centrosomes from vertebrate cellsCYTOSKELETON, Issue 4 2001A. Rousselet Abstract In an attempt to better understand the role of centrioles in vertebrate centrosomes, hydrostatic pressure was applied to isolated centrosomes as a means to disassemble centriole microtubules. Treatments of the centrosomes were monitored by analyzing their protein composition, ultrastructure, their ability to nucleate microtubules from pure tubulin, and their capability to induce parthenogenetic development of Xenopus eggs. Moderate hydrostatic pressure (95 MPa) already affected the organization of centriole microtubules in isolated centrosomes, and also impaired microtubule nucleation. At higher pressure, the protein composition of the peri-centriolar matrix (PCM) was also altered and the capacity to nucleate microtubules severely impaired. Incubation of the treated centrosomes in Xenopus egg extract could restore their capacity to nucleate microtubules after treatment at 95 MPa, but not after higher pressure treatment. However, the centriole structure was in no case restored. It is noteworthy that centrosomes treated with mild pressure did not allow parthenogenetic development after injection into Xenopus eggs, even if they had recovered their capacity to nucleate microtubules. This suggested that, in agreement with previous results, centrosomes in which centriole architecture is impaired, could not direct the biogenesis of new centrioles in Xenopus eggs. Centriole structure could also be affected by applying mild hydrostatic pressure directly to living cells. Comparison of the effect of hydrostatic pressure on cells at the G1/S border or on the corresponding cytoplasts suggests that pro-centrioles are very sensitive to pressure. However, cells can regrow a centriole after pressure-induced disassembly. In that case, centrosomes eventually recover an apparently normal duplication cycle although with some delay. Cell Motil. Cytoskeleton 48:262,276, 2001. © 2001 Wiley-Liss, Inc. [source] Panic and suicidal ideation and suicide attempts: results from the National Comorbidity SurveyDEPRESSION AND ANXIETY, Issue 3 2006M.P.H., Renee D. Goodwin Ph.D. Abstract Our objective was to determine the association between panic attacks (PAs) and panic disorder (PD), and suicidal ideation (SI) and suicide attempts (SAs) in a nationally representative sample of adults in the community. Data were drawn from the National Comorbidity Survey (n=5,877), a representative household sample of adults ages 15,54 in the United States. Multiple logistic regression analyses were used to examine the relationship between current and lifetime PA and PD and SI and SA, adjusting for differences in demographic characteristics, comorbid mental disorders (major depression, alcohol dependence, and substance dependence), childhood trauma (physical and sexual abuse), and number of lifetime mental disorders. Past-year and lifetime PA and PD were associated with increased SI (both past year and lifetime), and persisted after adjusting for comorbidity and early trauma. Associations between PA and SA were no longer statistically significant after adjusting for comorbidity. Past-year and lifetime PD were associated with lifetime SA, but these associations were no longer statistically significant after adjusting for comorbidity. Past-year and lifetime PD were associated with past-year SA, and this association persisted after adjusting for demographics, comorbidity, and number of lifetime mental disorders. These findings are consistent with previous results, and further help to clarify the relationships between panic and suicide behavior by identifying potential methodological reasons for inconsistencies in results from previous studies. Depression and Anxiety 23:124,132, 2006. © 2006 Wiley-Liss, Inc. [source] Alpha-melanocyte-stimulating hormone attenuates behavioral effects of corticotropin-releasing factor in isolated guinea pig pupsDEVELOPMENTAL PSYCHOBIOLOGY, Issue 5 2009Patricia A. Schiml-Webb Abstract During a 3-hr period of social isolation in a novel environment, guinea pig pups exhibit an initial active phase of behavioral responsiveness, characterized primarily by vocalizing, which is then followed by a stage of passive responsiveness in which pups display a distinctive crouch, eye-closing, and extensive piloerection. Prior treatment of pups with alpha-melanocyte-stimulating hormone (,-MSH) reduces each of the passive behaviors. The onset of passive responding during separation can be accelerated with peripheral injection of corticotropin-releasing factor (CRF). To examine whether CRF produces its effects through a mechanism similar to that of prolonged separation, we examined the effect of administering ,-MSH to pups injected with CRF. As expected, CRF markedly enhanced passive responding during a 60-min period of separation. ,-MSH delivered by either intracerebroventricular infusion or intraperitoneal injection significantly reduced each of the passive behavioral responses without significantly affecting active behavior. These findings, together with previous results indicating that it is the anti-inflammatory property of ,-MSH that is responsible for its behavioral effects during prolonged separation, suggest that peripheral CRF speeds the induction of passive responding through a mechanism involving enhanced proinflammatory activity. © 2009 Wiley Periodicals, Inc. Dev Psychobiol 51: 399,407, 2009. [source] Correlation model for spatially distributed ground-motion intensitiesEARTHQUAKE ENGINEERING AND STRUCTURAL DYNAMICS, Issue 15 2009Nirmal Jayaram Abstract Risk assessment of spatially distributed building portfolios or infrastructure systems requires quantification of the joint occurrence of ground-motion intensities at several sites, during the same earthquake. The ground-motion models that are used for site-specific hazard analysis do not provide information on the spatial correlation between ground-motion intensities, which is required for the joint prediction of intensities at multiple sites. Moreover, researchers who have previously computed these correlations using observed ground-motion recordings differ in their estimates of spatial correlation. In this paper, ground motions observed during seven past earthquakes are used to estimate correlations between spatially distributed spectral accelerations at various spectral periods. Geostatistical tools are used to quantify and express the observed correlations in a standard format. The estimated correlation model is also compared with previously published results, and apparent discrepancies among the previous results are explained. The analysis shows that the spatial correlation reduces with increasing separation between the sites of interest. The rate of decay of correlation typically decreases with increasing spectral acceleration period. At periods longer than 2,s, the correlations were similar for all the earthquake ground motions considered. At shorter periods, however, the correlations were found to be related to the local-site conditions (as indicated by site Vs30 values) at the ground-motion recording stations. The research work also investigates the assumption of isotropy used in developing the spatial correlation models. It is seen using the Northridge and Chi-Chi earthquake time histories that the isotropy assumption is reasonable at both long and short periods. Based on the factors identified as influencing the spatial correlation, a model is developed that can be used to select appropriate correlation estimates for use in practical risk assessment problems. Copyright © 2009 John Wiley & Sons, Ltd. [source] Representing genetic variation as continuous surfaces: an approach for identifying spatial dependency in landscape genetic studiesECOGRAPHY, Issue 6 2008Melanie A. Murphy Landscape genetics, an emerging field integrating landscape ecology and population genetics, has great potential to influence our understanding of habitat connectivity and distribution of organisms. Whereas typical population genetics studies summarize gene flow as pairwise measures between sampling localities, landscape characteristics that influence population genetic connectivity are often continuously distributed in space. Thus, there are currently gaps in both the ability to analyze genotypic data in a continuous spatial context and our knowledge of expected of landscape genetic structure under varying conditions. We present a framework for generating continuous "genetic surfaces", evaluate their statistical properties, and quantify statistical behavior of landscape genetic structure in a simple landscape. We simulated microsatellite genotypes under varying parameters (time since vicariance, migration, effective population size) and used ancestry (q) values from STRUCTURE to interpolate a genetic surface. Using a spatially adjusted Pearson's correlation coefficient to test the significance of landscape variable(s) on genetic structure we were able to detect landscape genetic structure on a contemporary time scale (,5 generations post vicariance, migration probability ,0.10) even when population differentiation was minimal (FST,0.00015). We show that genetic variation can be significantly correlated with geographic distance even when genetic structure is due to landscape variable(s), demonstrating the importance of testing landscape influence on genetic structure. Finally, we apply genetic surfacing to analyze an empirical dataset of black bears from northern Idaho USA. We find black bear genetic variation is a function of distance (autocorrelation) and habitat patch (spatial dependency), consistent with previous results indicating genetic variation was influenced by landscape by resistance. These results suggest genetic surfaces can be used to test competing hypotheses of the influence of landscape characteristics on genetic structure without delineation of categorical groups. [source] Effects of elevated CO2 associated with maize on multiple generations of the cotton bollworm, Helicoverpa armigeraENTOMOLOGIA EXPERIMENTALIS ET APPLICATA, Issue 1 2010Jin Yin Abstract Under elevated environmental carbon dioxide (CO2), leaf chewers tend to compensate for decreased leaf nutritional quality with increased consumption; mortality and development times also increase and cause a reduction in the fitness of leaf chewers. However, the effect of elevated CO2 on multiple successive generations of these and other insects is not well understood. Furthermore, information about the direct effects of increased environmental CO2 on developmental time and consumption of herbivores is lacking. In this paper, we tested the hypothesis that cascade effects of elevated CO2 through plants, rather than the direct effects of elevated CO2, are the main factors decreasing the fitness of cotton bollworm, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae). We used two series of experiments to quantify the growth, development, and consumption of H. armigera fed on an artificial diet or C4 plants (maize) grown under two CO2 levels (ambient vs. double ambient). In the first series of experiments, elevated CO2 had no effect on the population abundance or individual consumption for three successive generations of cotton bollworms fed on an artificial diet. In the second series of experiments, elevated CO2 reduced population abundance of cotton bollworm larvae for two successive generations when they were fed maize milky grains. The specific effects were longer larval duration, lower fecundity, and decreased rm of cotton bollworms. Furthermore, elevated CO2 increased individual consumption when cotton bollworm was fed maize milky grains for two successive generations and decreased the population's total consumption in the first generation but increased it in the second generation. The results from this study indicate that: (1) The effects of elevated CO2 on three successive generations of cotton bollworm fed on artificial diet were weak, or even non-existent, and (2) elevated CO2 increased the consumption when cotton bollworm were fed maize. Our study also suggests that the damage inflicted by cotton bollworm on maize (a C4 plant) will be seriously affected by the increases in atmospheric CO2, which is unlike our previous results for spring wheat (a C3 plant). [source] Sorption irreversibility of 1,4-dichlorobenzene in two natural organic matter,rich geosorbentsENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 3 2009Michael Sander Abstract Hysteresis, a frequently observed phenomenon in sorption studies, is inconsistent with the key assumption of sorption reversibility in most fate and bioavailability models. Therefore, a study of the underlying causes of hysteresis is essential. Carbon-radiolabeled 1,4-dichlorobenzene (DCB) isotope tracer exchange was carried out at select points along the isotherms of DCB in a brown coal and a peat soil, holding total DCB concentration constant. Tracer exchange was performed both in the forward (sorption) and reverse (desorption) directions at the bulk sorption points and in the desorption direction at the corresponding bulk desorption points. Bulk DCB isotherms showed concentration-dependent hysteresis. However, tracer reequilibration in all cases was consistent with free exchange between sorbed and aqueous-phase molecules. These results rule out common experimental artifacts and demonstrate that sorption of bulk DCB is truly hysteretic (i.e., irreversible). The differences in rates between bulk and tracer sorption and desorption are consistent with the coupling of bulk DCB diffusion to other processes that retard equilibration, which we assign to matrix swelling or shrinking. Hysteresis is attributed to matrix deformation,specifically, to inelastic expansion and creation of voids accommodating sorbate molecules in the matrix, which leads to enhanced affinity in the desorption step. Comparing the results to previous results for naphthalene in the coal, we find that irreversible effects are similar for DCB and naphthalene in the coal but differ for DCB between the two sorbents. An explanation based on the different physical properties of these sorbents is provided. Solid-phase extraction of equilibrated DCB with Tenax® revealed a highly desorption-resistant fraction. While too small to account for the observed hysteresis, this fraction may represent molecules that become trapped as the matrix collapses and simultaneously stiffens during abrupt desorption. [source] Ecological risk assessment of persistent toxic substances for the clam Tapes philipinarum in the lagoon of venice, italyENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 6 2004Christian Micheletti Abstract Because of contamination of sediments of the Lagoon of Venice, Italy, by inorganic pollutants (e.g., arsenic, cadmium, chromium, copper, lead, mercury, nickel, and zinc) and organic pollutants (e.g., polychlorobiphenyls), as well as the ecological and economical relevance of the edible clam Tapes philipinarum, an ecological risk assessment was undertaken to ascertain the extent of bioaccumulation that would pose a significant risk. Risk was estimated by means of toxic units and hazard quotient approaches, by comparing the exposure concentration with the effect concentration. Clam exposure was estimated by applying previous results based on bioaccumulation spatial regression models. In addition, a comparison was made between sum of dioxin-like polychlorinated biphenyl (PCB) congeners and total PCB bioaccumulation provided by spatial regression models and by a partitioning model. The effect concentrations were calculated as tissue screening concentrations, as the product of pollutant sediment quality criteria and the bioaccumulation factor. Finally, the cumulative risk posed by selected inorganic pollutants and total PCBs was estimated and a map of risk was drawn. The resulting chemicals of potential ecological concern were mercury, cadmium, arsenic, and nickel, as well as, to a lesser extent, total PCBs. [source] Saturday opening of alcohol retail shops in Sweden: an experiment in two phasesADDICTION, Issue 6 2005Thor Norström ABSTRACT Aim In February 2000, a trial started with Saturday opening of alcohol retail shops in certain parts of Sweden (phase I), and in July 2001, Saturday opening was extended to the whole country (phase II). The aim of this study is to assess the impact of phase II, and to probe previous results regarding phase I. Design Prior to February 2000, all alcohol monopoly outlets were closed on Saturdays. After this date, stores in an experimental area (six counties) were open on Saturdays. In the control area (seven counties) the shops remained closed. To prevent biases due to trade leakage, the experimental and control areas were separated by a buffer area (seven counties). Because continuous evaluations of the trial did not reveal any negative consequences, the Saturday opening was implemented in the whole of Sweden after 17 months. Data and methods The outcome measures included alcohol sales and indicators of assaults and drunk driving. The pre-intervention period covered the time period January 1995-January 2000, phase I of the post-intervention period February 2000,June 2001 (17 months), and phase II July 2001,July 2002 (13 months). The effects of the two phases were estimated through analyses of monthly data (auto-regressive integrated moving-average (ARIMA) modelling) depicting how sales and harm rates evolved in the experimental area compared to the control area during phase I as well as during phase II. Results The analysis uncovered a statistically significant increase in alcohol sales of 3.7% during phase I, and about the same increase during phase II (3.6%). There were no significant changes in any of the assault indicators, neither during phase I nor during phase II. There was a statistically significant increase in drunk driving (12%) during phase I, but no change during phase II. The analyses suggested that the increase during phase I was mainly due to a change in the surveillance strategy of the police. Conclusions The results lend support to the public health perspective in that the increased accessibility to alcohol rendered by Saturday opening also seems to have increased consumption. On the other hand, we could not detect any increase in alcohol-related harm. The question of whether this may be due to insufficient statistical power is discussed, together with some other methodological complications that were highlighted by the study. [source] Wilson's Storm Petrels Oceanites oceanicus Recognise the Olfactory Signature of Their MateETHOLOGY, Issue 12 2007Pierre Jouventin Chemical signals in birds have rarely been considered as recognition cues. Nevertheless, recent experiments showed that several petrel species are able to recognize their nest by smell, and in at least one species even their mate. But the use of smell may be different across the petrel species and olfactory nest recognition appears to be dependent on species' breeding biology. To increase our knowledge of individual olfactory recognition in petrels and the relationships between breeding biology and use of smell, we tested Wilson's storm petrels Oceanites oceanicus in Antarctica. In previous experiments, these birds failed to home if rendered anosmic, but the method employed to obtain anosmia (potentially stressing birds) and the fact that they breed in 24-h daylight suggest that they might use visual, rather than olfactory, cues to recognize their nest. Our birds were tested in T-maze experiments where nest odours or partner odours were presented. Wilson's storm petrels preferred odours of their own nest and mate. Results on olfactory nest recognition confirm and complete previous results, viz. anosmic Wilson's storm petrels do not home. Storm petrels olfactory mate recognition suggests that this ability may be widespread in burrowing petrels and implements olfactory nest recognition. [source] Strophe Length in Spontaneous Songs Predicts Male Response to Playback in the Hoopoe Upupa epopsETHOLOGY, Issue 5 2004Manuel Martín-Vivaldi Hoopoe (Upupa epops, Coraciformes) males produce a very simple song during the breeding season in order to attract females and repel intruders. Strophes vary in length (i.e. number of elements) both within and between males, and previous studies have shown that this song cue is positively correlated with male condition and breeding success. In the present study we tested whether strophe length of males influences male behaviour during intra-sexual contests, in a colour-ringed population in southeast Spain. Paired males were presented with a recorded song with long strophes during the pre-laying period, while they were near their mates, in order to provoke male mate-defence behaviour. Most males responded to the playback, but the strategy of defence adopted depended on their own strophe length in spontaneous songs recorded before the experiments. While singing responses were common to most of the males, only those using long strophes adopted the most risky strategy of approaching the loudspeaker. However, the males that approached produced abnormal songs during playback, that were shorter and with fewer strophes than those of males that did not approach, and used shorter strophes in comparison with spontaneous songs before the experiment. These differences in quality of the song produced in response to the playback suggest that long-strophe males were basing their response mainly on attacking rather than singing, while short-strophe males tried to resolve the contest at a distance by means of their song. These results show that strophe length reflects some component of the competitive ability of males (either physical strength or aggressiveness) in the hoopoe, which together with previous results regarding its role for female choice, show that it is a sexual signal with dual function. [source] | ||||||||||||||||||||||||||||||||||||||||||||||