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Selected AbstractsProtein kinase A RII-like (R2D2) proteins exhibit differential localization and AKAP interaction,CYTOSKELETON, Issue 7 2008Amy E. Hanlon Newell Abstract A-kinase anchoring proteins (AKAPs) bind to protein kinase A (PKA) via an amphipathic helix domain that interacts with a dimerization/docking domain on the regulatory (R) subunit of PKA. Four other mammalian proteins (ROPN1, ASP, SP17, and CABYR) also contain a highly conserved RII dimerization/docking (R2D2) domain, suggesting all four proteins may interact with all AKAPs in a manner similar to RII. All four of these proteins were originally detected in the flagellum of mammalian sperm. In this report, we demonstrate that all four R2D2 proteins are expressed in a wide variety of tissues and three of the proteins SP17, CABYR, and ASP are located in motile cilia of human bronchus and fallopian tubes. In addition, we detect SP17 in primary cilia. We also provide evidence that ROPN1 and ASP bind to a variety of AKAPs and this interaction can be disrupted with anchoring inhibitor peptides. The interaction of SP17 and CABYR with AKAPs appears to be much more limited. None of the R2D2 proteins appears to bind cAMP, a fundamental characteristic of the regulatory subunits of PKA. These observations suggest that R2D2 proteins utilize docking interactions with AKAPs to accomplish their function of regulating cilia and flagella. Based on location, affinity for AKAPs and lack of affinity for cAMP, it appears that each R2D2 protein has a unique role in this process. Cell Motil. Cytoskeleton 2008. © 2008 Wiley-Liss, Inc. [source] Functional analysis in Drosophila indicates that the NBCCS/PTCH1 mutation G509V results in activation of smoothened through a dominant-negative mechanismDEVELOPMENTAL DYNAMICS, Issue 4 2004Gary R. Hime Abstract Mutations in the human homolog of the patched gene are associated with the developmental (and cancer predisposition) condition Nevoid Basal Cell Carcinoma Syndrome (NBCCS), as well as with sporadic basal cell carcinomas. Most mutations that have been identified in the germline of NBCCS patients are truncating or frameshift mutations, with amino acid substitutions rarely found. We show that a missense mutation in the sterol-sensing domain G509V acts as a dominant negative when assayed in vivo in Drosophila. Ectopic expression of a Drosophila patched transgene, carrying the analogous mutation to G509V, causes ectopic activation of Hedgehog target genes and ectopic membrane stabilisation of Smoothened. The G509V transgene behaves in a manner similar, except in its subcellular distribution, to a C-terminal truncation that has been characterised previously as a dominant-negative protein. G509V exhibits vesicular localisation identical to the wild-type protein, but the C-terminal truncated Patched molecule is localised predominantly to the plasma membrane. This finding suggests that dominant-negative function can be conferred by interruption of different aspects of Patched protein behaviour. Another mutation at the same residue, G509R, did not exhibit dominant-negative activity, suggesting that simple removal of the glycine at 509 is not sufficient to impart dominant-negative function. Developmental Dynamics 229:780,790, 2004. © 2004 Wiley-Liss, Inc. [source] Polydivinylbenzene/Ethylvinylbenzene Composite Membranes for the Optimization of a Whole Blood Glucose SensorELECTROANALYSIS, Issue 1 2006Kerry Bridge Abstract A novel ultra thin polydivinylbenzene/ethylvinylbenzene composite membrane has been developed for use as the outer covering barrier in a model amperometric glucose oxidase enzyme electrode. The composite membrane was formed via the cathodic electropolymerization of divinylbenzene/ethylvinylbenzene at the surface of gold sputter coated host alumina membranes, (serving solely as a mechanical support for the thin polymer film). Permeability coefficients were determined for the enzyme substrates, O2 and glucose, across composite membranes formed with a range of polymer thicknesses. Due to the highly substrate diffusion limiting nature of the composite membrane, it was found that anionic interferents present in blood (such as ascorbate), were effectively screened from the working electrode via a charge exclusion mechanism, in a manner similar to previous findings within our laboratory. The enzyme electrode showed an initial 32% signal drift when first exposed to whole human blood over a period of 2 hours, after which time enzyme electrode responses remained essentially stable. Whole blood patient glucose determinations yielded a correlation coefficient of r2=0.97 in comparison to standard hospital analyses. [source] Toxicity assessment of reference and natural freshwater sediments with the LuminoTox assayENVIRONMENTAL TOXICOLOGY, Issue 4 2006P. M. Dellamatrice Abstract We examined the possibility of adapting the LuminoTox, a recently-commercialized bioanalytical testing procedure initially developed for aqueous samples, to assess the toxic potential of sediments. This portable fluorescent biosensor uses photosynthetic enzyme complexes (PECs) to rapidly measure photosynthetic efficiency. LuminoTox testing of 14 CRM (Certified Reference Material) sediments was first undertaken with (1) a "solid phase assay" (Lum-SPA) in which PECs are in intimate contact with sediment slurries for a 15 min exposure period and (2) an elutriate assay (Lum-ELU) in which PECs are exposed for 15 min to sediment water elutriates. CRM sediment toxicity data were then compared with those generated with the Microtox Solid Phase Assay (Mic-SPA). A significant correlation (P < 0.05) was shown to exist between Lum-SPA and Mic-SPA, indicating that both tests display a similar toxicity response pattern for CRM sediments having differing contaminant profiles. The sediment elutriate Lum-ELU assay displayed toxicity responses (i.e. measurable IC20s) for eight of the 14 CRM sediments, suggesting that it is capable of determining the presence of sediment contaminants that are readily soluble in an aqueous elutriate. Lum-SPA and Mic-SPA bioassays were further conducted on 12 natural freshwater sediments and their toxicity responses were more weakly, yet significantly, correlated. Finally, Lum-SPA testing undertaken with increasing mixtures of kaolin clay confirmed that its toxicity responses, in a manner similar to those reported for the Mic-SPA assay, are also subject to the influence of grain size. While further studies will be required to more fully understand the relationship between Lum-SPA assay responses and the physicochemical makeup of sediments (e.g., grain size, combined presence of natural and anthropogenic contaminants), these preliminary results suggest that LuminoTox testing could be a useful screen to assess the toxic potential of solid media. © 2006 Wiley Periodicals, Inc. Environ Toxicol 21: 395,402, 2006. [source] Toxicity of manufactured zinc oxide nanoparticles in the nematode Caenorhabditis elegansENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 6 2009Hongbo Ma Abstract Information describing the possible impacts of manufactured nanoparticles on human health and ecological receptors is limited. The objective of the present study was to evaluate the potential toxicological effects of manufactured zinc oxide nanoparticles (ZnO-NPs; 1.5 nm) compared to aqueous zinc chloride (ZnCl2) in the free-living nematode Caenorhabditis elegans. Toxicity of both types of Zn was investigated using the ecologically relevant endpoints of lethality, behavior, reproduction, and transgene expression in a mtl-2::GFP (gene encoding green fluorescence protein fused onto the metallothionein-2 gene promoter) transgenic strain of C. elegans. Zinc oxide nanoparticles showed no significant difference from ZnCl2 regarding either lethality or reproduction in C. elegans, as indicated by their median lethal concentrations (LC50s; p = 0.29, n = 3) and median effective concentrations (EC50s; Z = 0.835, p = 0.797). Also, no significant difference was found in EC50s for behavioral change between ZnO-NPs (635 mg Zn/L; 95% confidence interval [CI], 477,844 mg Zn/L) and ZnCl2 (546 mg Zn/L; 95% CI, 447,666 mg Zn/L) (Z = 0.907, p = 0.834). Zinc oxide nanoparticles induced transgene expression in the mtl-2::GFP transgenic C. elegans in a manner similar to that of ZnCl2, suggesting that intracellular biotransformation of the nanoparticles might have occurred or the nanoparticles have dissolved to Zn2+ to enact toxicity. These findings demonstrate that manufactured ZnO-NPs have toxicity to the nematode C. elegans similar to that of aqueous ZnCl2. [source] The Prestige oil spill.ENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 9 2005Abstract In vitro biodegradation of the Prestige heavy fuel oil has been carried out using two microbial consortia obtained by enrichment in different substrates to simulate its environmental fate and potential utility for bioremediation. Different conditions, such as incubation time (i.e., 20 or 40 d), oil weathering, and addition of an oleophilic fertilizer (S200), were evaluated. Weathering slowed down the degradation of the fuel oil, probably because of the loss of lower and more labile components, but the addition of S200 enhanced significantly the extension of the biodegradation. n -Alkanes, alkylcyclohexanes, alkylbenzenes, and the two- to three-ring polycyclic aromatic hydrocarbons (PAHs) were degraded in 20 or 40 d of incubation of the original oil, whereas the biodegradation efficiency decreased for higher PAHs and with the increase of alkylation. Molecular markers were degraded according to the following sequence: Acyclic isoprenoids < diasteranes < C27 -steranes < ,,-steranes < homohopanes < monoaromatic steranes < triaromatic steranes. Isomeric selectivity was observed within the C1 - and C2 -phenanthrenes, dibenzothiophenes, pyrenes, and chrysenes, providing source and weathering indices for the characterization of the heavy oil spill. Acyclic isoprenoids, C27 -steranes, C1 - and C2 -naphthalenes, phenanthrenes, and dibenzothiophenes were degraded completely when S200 was used. The ratios of the C2 - and C3 -alkyl homologues of fluoranthene/pyrene and chrysene/benzo[a]anthracene are proposed as source ratios in moderately degraded oils. The 4-methylpyrene and 3-methylchrysene were refractory enough to serve as conserved internal markers in assessing the degradation of the aromatic fraction in a manner similar to that of hopane, as used for the aliphatic fraction. [source] Plasticity of hair follicle dermal cells in wound healing and inductionEXPERIMENTAL DERMATOLOGY, Issue 2 2003A. Gharzi Abstract: The capacity of adult hair follicle dermal cells to participate in new follicle induction and regeneration, and to elicit responses from diverse epithelial partners, demonstrates a level of developmental promiscuity and influence far exceeding that of interfollicular fibroblasts. We have recently suggested that adult follicle dermal cells have extensive stem or progenitor cell activities, including an important role in skin dermal wound healing. Given that up to now tissue engineered skin equivalents have several deficiencies, including the absence of hair follicles, we investigated the capacity of follicle dermal cells to be incorporated into skin wounds; to form hair follicles in wound environments; and to create a hair follicle-derived skin equivalent. In our study, we implanted rat follicle dermal cells labelled with a vital dye into ear and body skin wounds. We found that they were incorporated into the new dermis in a manner similar to skin fibroblasts, but that lower follicle dermal sheath also assimilated into hair follicles. Using different combinations of follicle dermal cells and outer root sheath epithelial cells in punch biopsy wounds, we showed that new hair follicles were formed only with the inclusion of intact dermal papillae. Finally by combining follicle dermal sheath and outer root sheath cells in organotypic chambers, we created a skin equivalent with characteristic dermal and epidermal architecture and a normal basement membrane , the first skin to be produced entirely from hair follicle cells. These data support the hypothesis that follicle dermal cells may be important in wound healing and demonstrate their potential usefulness in human skin equivalents and skin substitutes. While we have made progress towards producing skin equivalents that contain follicles, we suggest that the failure of cultured dermal papilla cells to induce follicle formation in wounds illustrates the complex role the follicle dermis may play in skin. We believe that it demonstrates a genuine dichotomy of activity for follicle cells within skin. [source] Experimental and numerical studies on dynamic crack growth in layered slate rock under wedge impact loads: part II , non-plane strain problemFATIGUE & FRACTURE OF ENGINEERING MATERIALS AND STRUCTURES, Issue 10 2007M. R. ALAM ABSTRACT Dynamic crack propagation in non-plane strain (or 3D) slate blocks under wedge impact loads was investigated numerically in this part of the paper. A parabolic-shaped crack trajectory was taken into consideration to model the crack propagation in slate blocks for analyzing the impact splitting of layered slate rock. Major and minor axes of the parabola were determined from the condition of equal mode I stress intensity factors (SIFs) along the crack front. Mode I SIFs were determined for experimental breaking loads for each increment of crack growth in a manner similar to that mentioned in part I of this paper. These values were compared with the plane strain material fracture toughness value obtained from experimental studies and very good agreement was obtained between them, for the case of actual load applied on the specimen. Numerical analysis of a field problem, i.e., separation of a large-sized slate slab from the rock strata in a slate quarry using wedge impacting, was also carried out in this paper. It can be observed that a large magnitude of load is required to break large-sized slate blocks; but this load is applied through a number of smaller load-capacity actuators-in-parallel, requiring large power capacity for the hydraulic pumps. However, this required power could be reduced considerably if the load applied on the line of hydraulic actuators is cascaded across the (line of) actuators (starting from centrally placed actuators) with a small time delay (equal to the initial crushing time in slate rock). [source] Identification of an osteopontin-like protein in fish associated with mineral formationFEBS JOURNAL, Issue 17 2007Vera G. Fonseca Fish has been recently recognized as a suitable vertebrate model and represents a promising alternative to mammals for studying mechanisms of tissue mineralization and unravelling specific questions related to vertebrate bone formation. The recently developed Sparus aurata (gilthead seabream) osteoblast-like cell line VSa16 was used to construct a cDNA subtractive library aimed at the identification of genes associated with fish tissue mineralization. Suppression subtractive hybridization, combined with mirror orientation selection, identified 194 cDNA clones representing 20 different genes up-regulated during the mineralization of the VSa16 extracellular matrix. One of these genes accounted for 69% of the total number of clones obtained and was later identified as theS. aurata osteopontin-like gene. The 2138-bp full-length S. aurata osteopontin-like cDNA was shown to encode a 374 amino-acid protein containing domains and motifs characteristic of osteopontins, such as an integrin receptor-binding RGD motif, a negatively charged domain and numerous post-translational modifications (e.g. phosphorylations and glycosylations). The common origin of mammalian osteopontin and fish osteopontin-like proteins was indicated through an in silico analysis of available sequences showing similar gene and protein structures and was further demonstrated by their specific expression in mineralized tissues and cell cultures. Accordingly, and given its proven association with mineral formation and its characteristic protein domains, we propose that the fish osteopontin-like protein may play a role in hard tissue mineralization, in a manner similar to osteopontin in higher vertebrates. [source] Biophysical characterization of the interaction of Limulus polyphemus endotoxin neutralizing protein with lipopolysaccharideFEBS JOURNAL, Issue 10 2004Jörg Andrä Endotoxin-neutralizing protein (ENP) of the horseshoe crab is one of the most potent neutralizers of endotoxins [bacterial lipopolysaccharide (LPS)]. Here, we report on the interaction of LPS with recombinant ENP using a variety of physical and biological techniques. In biological assays (Limulus amebocyte lysate and tumour necrosis factor-, induction in human mononuclear cells), ENP causes a strong reduction of the immunostimulatory ability of LPS in a dose-dependent manner. Concomitantly, the accessible negative surface charges of LPS and lipid A (zeta potential) are neutralized and even converted into positive values. The gel to liquid crystalline phase transitions of LPS and lipid A shift to higher temperatures indicative of a rigidification of the acyl chains, however, the only slight enhancement of the transition enthalpy indicates that the hydrophobic moiety is not strongly disturbed. The aggregate structure of lipid A is converted from a cubic into a multilamellar phase upon ENP binding, whereas the secondary structure of ENP does not change due to the interaction with LPS. ENP contains a hydrophobic binding site to which the dye 1-anilino-8-sulfonic acid binds at a Kd of 19 µm, which is displaced by LPS. Because lipopolysaccharide-binding protein (LBP) is not able to bind to LPS when ENP and LPS are preincubated, tight binding of ENP to LPS can be deduced with a Kd in the low nonomolar range. Importantly, ENP is able to incorporate by itself into target phospholipid liposomes, and is also able to mediate the intercalation of LPS into the liposomes thus acting as a transport protein in a manner similar to LBP. Thus, LPS,ENP complexes might enter target membranes of immunocompetent cells, but are not able to activate due to the ability of ENP to change LPS aggregates from an active into an inactive form. [source] Group IID heparin-binding secretory phospholipase A2 is expressed in human colon carcinoma cells and human mast cells and up-regulated in mouse inflammatory tissuesFEBS JOURNAL, Issue 11 2002Makoto Murakami Group IID secretory phospholipase A2 (sPLA2 -IID), a heparin-binding sPLA2 that is closely related to sPLA2 -IIA, augments stimulus-induced cellular arachidonate release in a manner similar to sPLA2 -IIA. Here we identified the residues of sPLA2 -IID that are responsible for heparanoid binding, are and therefore essential for cellular function. Mutating four cationic residues in the C-terminal portion of sPLA2 -IID resulted in abolition of its ability to associate with cell surface heparan sulfate and to enhance stimulus-induced delayed arachidonate release, cyclooxygenase-2 induction, and prostaglandin generation in 293 cell transfectants. As compared with several other group II subfamily sPLA2s, which were equally active on A23187- and IL-1-primed cellular membranes, sPLA2 -IID showed apparent preference for A23187-primed membranes. Several human colon carcinoma cell lines expressed sPLA2 -IID and sPLA2 -X constitutively, the former of which was negatively regulated by IL-1. sPLA2 -IID, but not other sPLA2 isozymes, was expressed in human cord blood-derived mast cells. The expression of sPLA2 -IID was significantly altered in several tissues of mice with experimental inflammation. These results indicate that sPLA2 -IID may be involved in inflammation in cell- and tissue-specific manners under particular conditions. [source] Effects of catch and release angling on Atlantic salmon, Salmo salar L., of the Conne River, NewfoundlandFISHERIES MANAGEMENT & ECOLOGY, Issue 3 2002J. B. DEMPSON The effects of catch and release angling on survival of Atlantic salmon, Salmo salar L., at Conne River, Newfoundland, were investigated by retaining angled (n=49; experimental group) and trap-caught (n=20; control group) fish in holding cages for up to 40 days. Samples were obtained from 8 June to 4 July, 2000, and partitioned among four water temperature strata. Apart from not being angled, control fish were handled, tagged, and transferred to holding cages in a manner similar to angled salmon. Water temperatures and discharge were monitored throughout the duration of the study. Overall, 8.2% of salmon caught and released died, but 12% died among salmon angled in water temperatures , 17.9 °C. No control fish died. There were no significant differences in time associated with angling, exposure to air, tagging, transfer to holding cages, nor total handling time between salmon that survived vs. those that died. Results of the study should encourage managers to continue to use catch and release as a viable tool in the management of Atlantic salmon stocks. [source] Thermal performance of juvenile Atlantic Salmon, Salmo salar L.FUNCTIONAL ECOLOGY, Issue 6 2001B. JONSSON Summary 1,Experimental data for maximum growth and food consumption of Atlantic Salmon (Salmo salar L.) parr from five Norwegian rivers situated between 59 and 70°N were analysed and modelled. The growth and feeding models were also applied to groups of Atlantic Salmon growing and feeding at rates below the maximum. The data were fitted to the Ratkowsky model, originally developed for bacterial growth. 2,The rates of growth and food consumption varied significantly among populations but the variation appeared unrelated to thermal conditions in the river of population origins. No correlation was found between the thermal conditions and limits for growth, thermal growth optima or maximum growth, and hypotheses of population-specific thermal adaptation were not supported. Estimated optimum temperatures for growth were between 16 and 20 °C. 3, Model parameter estimates differed among growth-groups in that maximum growth and the performance breadth decreased from fast to slow growing individuals. The optimum temperature for growth did not change with growth rate. 4, The model for food consumption (expressed in energy terms) peaked at 19,21 °C, which is only slightly higher than the optimal temperature for growth. Growth appeared directly related to food consumption. Consumption was initiated ,2 °C below the lower temperature for growth and terminated ,1·5 °C above the upper critical temperature for growth. Model parameter estimates for consumption differed among growth-groups in a manner similar to the growth models. 5,By combining the growth and consumption models, growth efficiencies were estimated. The maximum efficiencies were high, 42,58%, and higher in rivers offering hostile than benign feeding and growth opportunities. [source] Focal nodular hyperplasia: what are the indications for resection?HPB, Issue 4 2005Li Chun Hsee Focal nodular hyperplasia (FNH) is a benign condition of the liver that is often discovered incidentally on radiological investigation. FNH has no malignant potential, is rarely symptomatic and surgical intervention is almost never required. However, eight patients with a diagnosis of FNH associated with upper abdominal pain or rapid growth were referred for surgery. All patients had been extensively investigated for other causes of pain and had been observed for between 1 and 7 years prior to surgical referral. The FNH lesions were between 1 cm and 8 cm in diameter. One FNH lesion 7.5 cm in diameter lay in segment VII/VIII and was related to the right and middle hepatic veins. All patients were resected with immediate and lasting control of their symptoms. Based on this experience FNH should be managed in a manner similar to haemangiomas with most lesions being safe to observe. However, it should be recognized that symptomatic FNH does occur, as well as FNH behaving in an unusual fashion such as rapid growth. Both of these findings are indications for resection. [source] Determination of haplotypes from single DNA molecules: a method for single-molecule barcoding,,HUMAN MUTATION, Issue 9 2007Ming Xiao Abstract Determining the haplotypes in a diploid individual is a major technical challenge in genetic studies of human complex traits. Here we report a method of molecular haplotyping by directly imaging multiple polymorphic sites on individual DNA molecules simultaneously. DNA fragments amplified by long-range PCR were labeled with fluorescent dyes at each polymorphic site using a modified gap-filled padlock probe ligation approach. The labeled DNA molecules were then stretched into linear form on a functionalized glass surface and imaged with multicolor total internal reflection fluorescence (TIRF) microscopy. By determining the colors and positions of the fluorescent labels with respect to the backbone at polymorphic sites, the haplotype can be inferred accurately, in a manner similar to reading a barcode, even when the DNA fragments are not fully labeled. The feasibility of this technology is demonstrated by the determination of the haplotype of a 9.3-kbp DNA fragment containing four SNPs. Hum Mutat 28(9), 913,921, 2007. Published 2007 Wiley-Liss, Inc. [source] Ascochlorin activates p53 in a manner distinct from DNA damaging agentsINTERNATIONAL JOURNAL OF CANCER, Issue 12 2009Ji-Hak Jeong Abstract Ascochlorin, a prenylphenol antitumor antibiotic, profoundly increases the expression of endogenous p53 by increasing protein stability in the human osteosarcoma cells and human colon cancer cells. Ascochlorin also increases DNA binding activity to the p53 consensus sequence in nuclear extract and enhances transcription of p53 downstream targets. Ascochlorin specifically induces p53 phosphorylation at ser 392 without affecting ser 15 or 20, whereas DNA damaging agents typically phosphorylate these serines. Moreover, ascochlorin does not induce phosphorylation of ATM and CHK1, an established substrate of ATR that is activated by genotoxins, nor does it increase DNA strand break, as confirmed by comet assay. The structure-activity relationship suggests that p53 activation by ascochlorin is related to inhibition of mitochondrial respiration, which is further supported by the observation that respiratory inhibitors activate p53 in a manner similar to ascochlorin. These results suggest that ascochlorin, through the inhibition of mitochondrial respiration, activates p53 through a mechanism distinct from genotoxins. © 2009 UICC [source] Superresolution planar diffraction tomography through evanescent fields,INTERNATIONAL JOURNAL OF IMAGING SYSTEMS AND TECHNOLOGY, Issue 1 2002Sean K. Lehman We consider the problem of noninvasively locating objects buried in a layered medium such as land mines in the ground or objects concealed in a wall. In such environments, the transmitter(s) and receiver(s) are frequently within the near-field region of the illuminating radiation. In these cases, the scattered evanescent field carries useful information on the scattering object. Conventional diffraction tomography techniques neglect, by their design, the evanescent field. Under near-field conditions, they treat it as noise as opposed to valid data. If correctly incorporated into a reconstruction algorithm, the evanescent field, which carries high spatial frequency information, can be used to achieve resolution beyond the classical limit of ,/2, or "superresolution." We build on the generalized holography theory presented by Langenberg to develop a planar diffraction tomography algorithm that incorporates evanescent field information to achieve superresolution. Our theory is based on a generalization of the Fourier transform, which allows for complex spatial frequencies in a manner similar to the Laplace transform. We specialize our model to the case of a two-dimensional multimonostatic, wideband imaging system, and derive an extended resolution reconstruction procedure. We implement and apply our reconstruction to two data sets collected using the Lawrence Livermore National Laboratory (LLNL) Micropower Impulse Radar (MIR). © 2002 John Wiley & Sons, Inc. Int J Imaging Syst Technol 12, 16,26, 2002 [source] New perspectives on the fundamental theorem of density functional theoryINTERNATIONAL JOURNAL OF QUANTUM CHEMISTRY, Issue 15 2008Xiao-Yin Pan Abstract The fundamental theorem of time-independent/time-dependent density functional theory due to Hohenberg,Kohn (HK)/Runge,Gross (RG) proves the bijectivity between the density ,(r)/,(rt) and the Hamiltonian /(t) to within a constant C/function C(t), and wave function ,/, (t). The theorems are each proved for scalar external potential energy operators. By a unitary or equivalently a gauge transformation that preserves the density, we generalize the realm of validity of each theorem to Hamiltonians, which additionally include the momentum operator and a curl-free vector potential energy operator defined in terms of a gauge function , (R)/, (Rt). The original HK/RG theorems then each constitute a special case of this generalization. Thereby, a fourfold hierarchy of such theorems is established. As a consequence of the generalization, the wave function ,/, (t) is shown to be a functional of both the density ,(r)/,(rt), which is a gauge-invariant property, and a gauge function ,(R)/,(Rt). The functional dependence on the gauge function ensures that as required by quantum mechanics, the wave function written as a functional is gauge variant. The hierarchy and the dependence of the wave function functional on the gauge function thus enhance the significance of the phase factor in density functional theory in a manner similar to that of quantum mechanics. Various additional perspectives on the theorem are arrived at. These understandings also address past critiques of time-dependent theory. © 2008 Wiley Periodicals, Inc. Int J Quantum Chem, 2008 [source] The sperm nuclear matrix is required for paternal DNA replication,JOURNAL OF CELLULAR BIOCHEMISTRY, Issue 3 2007Jeffrey A. Shaman Abstract The mammalian sperm nucleus provides an excellent model for studying the relationship between the formation of nuclear structure and the initiation of DNA replication. We previously demonstrated that mammalian sperm nuclei contain a nuclear matrix that organizes the DNA into loop domains in a manner similar to that of somatic cells. In this study, we tested the minimal components of the sperm nucleus that are necessary for the formation of the male pronucleus and for the initiation of DNA synthesis. We extracted mouse sperm nuclei with high salt and dithiothreitol to remove the protamines in order to form nuclear halos. These were then treated with either restriction endonucleases to release the DNA not directly associated with the nuclear matrix or with DNAse I to digest all the DNA. The treated sperm nuclei were injected into oocytes, and the paternal pronuclear formation and DNA synthesis was monitored. We found that restriction digested sperm nuclear halos were capable of forming paternal pronuclei and initiating DNA synthesis. However, when isolated mouse sperm DNA or sperm DNA reconstituted with the nuclear matrices were injected into oocytes, no paternal pronuclear formation or DNA synthesis was observed. These data suggest that the in situ nuclear matrix attachment organization of sperm DNA is required for mouse paternal pronuclear DNA synthesis. J. Cell. Biochem. 102: 680,688, 2007. © 2007 Wiley-Liss, Inc. [source] Service quality in agronomic inputs: does the hierarchical model apply?AGRIBUSINESS : AN INTERNATIONAL JOURNAL, Issue 4 2009Michael A. Gunderson Research regarding the quality of services is increasingly important as services account for more of the national economy. In agriculture many services are used, but are not systematically assessed in a manner similar to how products are assessed. This research aims to identify the compatibility of proposed service quality structures to the agriculture inputs sector. Results of a survey of Midwest row crop producers lend support to the hierarchical model of service quality structure. Input suppliers can use these results to structure their services more efficiently. [EconLit classification codes: M, Q]. © 2009 Wiley Periodicals, Inc. [source] Field performance and seasonal changes in the efficacy against Helicoverpa armigera (Hübner) of transgenic cotton expressing the insecticidal protein vip3AAGRICULTURAL AND FOREST ENTOMOLOGY, Issue 2 2007Danny J. Llewellyn Abstract 1,Three years of field experiments in Eastern Australia were carried out on transgenic cotton (Gossypium hirsutum L.) event Cot102 expressing the insecticidal protein gene vip3A from Bacillus thuringiensis to evaluate performance against Helicoverpa armigera Hübner. Efficacy, defined as the capacity of plant tissues to induce larval mortality, was determined with a well-validated leaf bioassay fortnightly through the growth cycle of the cotton in each season. 2,Cot102 plants proved highly efficacious against H. armigera, particularly early in the season, although their efficacy declined as the season progressed, in a manner similar to, but not as dramatic as, that observed with commercial Cry1Ac expressing cotton (Bollgard or Ingard cotton). 3,Field surveys indicated that very few larvae survived beyond first instar on intact growing plants. 4,In one season efficacy declined for a period of approximately 20 days after a cool wet period, suggesting that this may have had a detrimental effect on the expression or efficacy of the gene, but this will need to be verified in further replicated trials. 5,Quantitative enzyme-linked immunosorbent assays indicated that there was no dramatic reduction in production of the vip3A protein during growth and maturation of the crop, suggesting that other host plant factors were affecting the efficacy of the insecticidal protein in the insect gut. 6,These data indicate that Cot102 cotton would provide a useful alternative to Bollgard cotton but, given the similar lytic mode of action of vip3A proteins in the insect midgut, there may be similar inherent vulnerabilities to resistance evolution for these proteins if used alone. Pyramiding of the vip3A trait with a second insecticidal gene would appear to be a high priority for achieving sustainable deployment against H. armigera or similar susceptible species. [source] Some Preparation Required: The Journey To Successful Studio CollaborationJOURNAL OF INTERIOR DESIGN, Issue 2 2006Jennifer D. Webb Ph.D. ABSTRACT The purpose of this study was to explore relationships between team training perceptions and training effectiveness among students and faculty. The three objectives for this project were: 1) compare student and faculty perceptions concerning the provision of team training in design studios; 2) investigate the relationship between students' reported team training levels and students' reported team attitudes; and 3) investigate the relationship between students' reported team training levels and students' reported team behaviors. IDEC members and their interior design students were selected to participate in the study. Questionnaires were mailed to the faculty and the instruments were administered in a manner similar to course evaluations. Chi Square analysis suggests that instructors were more likely than students to indicate that they had provided training on effective communication, task division, conflict resolution, and characteristics of a good team. The findings suggest that teamwork training is positively related to positive perceptions and attitudes about teamwork, and to proactive behaviors in team settings. Most importantly, findings indicate that participation in multiple team projects is not related to improved attitudes or behaviors. This result emphasizes the role of preparation in successful studio collaboration. [source] "Revenue Accounting" in the Age of E-Commerce: A Framework for Conceptual, Analytical, and Exchange Rate ConsiderationsJOURNAL OF INTERNATIONAL FINANCIAL MANAGEMENT & ACCOUNTING, Issue 1 2002Jonathan C. Glover This paper explores "revenue accounting" in contrast to traditional "cost accounting". Revenue accounting serves the information needs of managers and investors in planning and controlling a firm's sales activities and their financial consequences, especially in the age of e-commerce. Weaknesses of traditional accounting have become particularly evident recently, for example, the lack of 1) revenue mileposts, 2) revenue sustainability measurements, and 3) intangibles capitalization. The paper emphasizes the need to develop a conceptual framework of revenue accounting and, as a tentative measure, proposes five basic postulates and five operational postulates of revenue accounting. On the side of analytical frameworks, the paper explores some tentative remedies for the weaknesses. Several revenue mileposts are explored to gauge progress in earning revenues and a Markov process is applied to an example involving mileposts. Revenue momentum, measured by the exponential smoothing method, is examined as a way of getting feedback on revenue sustainability; and the use of the sustainability concept in the analysis of "fixed and variable revenues" is illustrated. A project-oriented approach in a manner similar to capital budgeting and to Reserve Recognition Accounting is proposed by treating each customer as a project. Standardization of forecasts are also considered as an important way of bypassing the capitalization issue. Finally, while e-commerce is inherently global, issues specific to global operations are highlighted, namely, exchange rate issues when venture capitalists and the start-up company use different currencies producing different rates of return on the same project. [source] Regulation of glial development by cystatin CJOURNAL OF NEUROCHEMISTRY, Issue 1 2007Akiko Hasegawa Abstract Cystatin C (CysC) is an endogenous cysteine proteases inhibitor produced by mature astrocytes in the adult brain. Previously we isolated CysC as a factor activating the glial fibrillary acidic protein (GFAP) promoter, and showed that CysC is expressed in astrocyte progenitors during development. Here we show that protease inhibitor activity increased daily in conditioned medium, and that this activity was mainly a result of CysC released from primary cultured cells. Human CysC added to the culture medium of primary brain cells increased the number of GFAP-positive and nestin-positive cells. Human CysC also increased the number of neurospheres formed from embryonic brain, and thus it increases the number of neural stem/precursor cells in a manner similar to glycosylated rat CysC. The addition of a neutralizing antibody, on the other hand, greatly decreased the number of GFAP and glutamate aspartate transporter (GLAST)-positive astrocytes. This decrease was reversed by the addition of CysC but not by another cysteine protease inhibitor. Thus, the promotion of astrocyte development by CysC appears to be independent of its protease inhibitor activity. The antibody increased the number of oligodendrocytes and their precursors. Therefore, CysC modifies glial development in addition to its activity against neural stem/precursor cells. [source] Identification of a new functional target of haloperidol metabolite: implications for a receptor-independent role of 3-(4-fluorobenzoyl) propionic acidJOURNAL OF NEUROCHEMISTRY, Issue 2 2006Hyeon Soo Kim Abstract Haloperidol, a dopamine D2 receptor blocker, is a classical neuroleptic drug that elicits extrapyramidal symptoms. Its metabolites include 3-(4-fluorobenzoyl) propionic acid (FBPA) and 4-(4-chlorophenyl)-4-piperidinol (CPHP). Until now, the biological significance of these metabolites has remained largely unknown. Here, we report that the administration of FBPA to mice effected a suppression of locomotor activity and induced catalepsy in a manner similar to that observed with haloperidol, whereas CPHP had no significant effects. Neither of these two metabolites, however, exhibited any ability to bind to the dopamine D2 receptor. FBPA blocked dopamine-induced extracellular signal-regulated kinase 1/2 phosphorylation, and it specifically affected mitogen-activated protein kinase kinase (MEK)1/2 activity in hippocampal HN33 cells. Moreover, FBPA was capable of direct interaction with MEK1/2, and inhibited its activity in vitro. We demonstrated the generation of haloperidol metabolites within haloperidol-treated cells by mass spectrometric analyses. Collectively, our results confirm the biological activity of FBPA, and provide initial clues as to the receptor-independent role of haloperidol. [source] Multilineage mesenchymal differentiation potential of human trabecular bone-derived cellsJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 5 2002Ulrich Nöth Abstract Explant cultures of adult human trabecular bone fragments give rise to osteoblastic cells, that are known to express osteoblast-related genes and mineralize extracellular matrix. These osteoblastic cells have also been shown to undergo adipogenesis in vitro and chondrogenesis in vivo. Here we report the in vitro developmental potential of adult human osteoblastic cells (hOB) derived from explant cultures of collagenase-pretreated trabecular bone fragments. In addition to osteogenic and adipogenic differentiation, these cells are capable of chondrogenic differentiation in vitro in a manner similar to adult human bone marrow-derived mesenchymal progenitor cells. High-density pellet cultures of hOB maintained in chemically defined serum-free medium, supplemented with transforming growth factor-,1, were composed of morphologically distinct, chondrocyte-like cells expressing mRNA transcripts of collagen types II, IX and X, and aggrecan. The cells within the high-density pellet cultures were surrounded by a sulfated prote-oglycan-rich extracellular matrix that immunostained for collagen type II and proteoglycan link protein. Osteogenic differentiation of hOB was verified by an increased number of alkaline phosphatase-positive cells, that expressed osteoblast-related transcripts such as alkaline phosphatase, collagen type I, osteopontin and osteocalcin, and formed mineralized matrix in monolayer cultures treated with ascorbate, ,-glycerophosphate, and bone morphogenetic protein-2. Adipogenic differentiation of hOB was determined by the appearance of intracellular lipid droplets, and expression of adipocyte-specific genes, such as lipoprotein lipase and peroxisome proliferator-activated receptor ,2, in monolayer cultures treated with dexamethasone, indomethacin, insulin and 3-isobutyl-l-methylxanthine. Taken together, these results show that cells derived from collagenase-treated adult human trabecular bone fragments have the potential to differentiate into multiple mesenchymal lineages in vitro, indicating their developmental plasticity and suggesting their mesenchymal progenitor nature. © 2002 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved. [source] Pharmacokinetics of the sequential metabolites of loteprednol etabonate in ratsJOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 3 2008Whei-Mei Wu Pharmacokinetics, metabolism and excretion of two sequential inactive metabolites of the soft corticosteroid loteprednol etabonate (LE), ,1 -cortienic acid etabonate (AE) and ,1 -cortienic acid (A), have been investigated in rats. Pharmacokinetic studies (two-compartment model, 10 mg kg,1 intravenous bolus of AE or A) found the elimination of both AE (t1/2(,), 12.46 ± 1.18 min; CLtotal, 101.94 ± 5.80 mL min,1 kg,1; and Kel, 0.24 ± 0.02 min,1) and A (t1/2(,), 14.62 ± 0.46 min; CLtotal, 53.80 ± 1.40 mL min,1 kg,1; and Kel, 0.18 ± 0.02 min,1) to be significantly faster than that previously determined for the parent LE (t1/2(,), 43.41 ± 7.58 min; CLtotal, 67.40 ± 11.60 mL min,1 kg,1; and Kel, 0.071 ± 0.024 min,1). For metabolism and excretion evaluations, 1 and 10 mg kg,1 of either AE or A were intravenously administered, and the urine and bile were collected. AE and A rapidly reached their peak concentrations in the bile and urine, and most of them were eliminated within one hour. Total cumulative excretions at 4 h after 1 and 10 mg kg,1 injections were 85.51 ± 3.38% and 67.50 ± 2.67% for AE, and 71.90 ± 3.72% and 37.73 ± 2.69% for A in bile; and 4.84 ± 1.87% and 13.85 ± 3.27% for AE, and 24.28 ± 8.44% and 22.35 ± 1.12% for A in urine, respectively. After AE administration, the excretion of AE was > 90%, and A was < 10% in all cases, indicating that the elimination of AE was much faster than its metabolism (to A). In a manner similar to that seen for LE, dose-dependent elimination was observed both in AE and A. These results suggested that both AE and A were ideal leads for the design of soft steroids based on the inactive metabolite approach. [source] African-American Developmental Disability Discourses: Implications for Policy DevelopmentJOURNAL OF POLICY AND PRACTICE IN INTELLECTUAL DISABILITIES, Issue 1 2005Peggy S. Terhune Abstract, Formal developmental disabilities services are often underutilized, especially by low-income African-American families. This study examined the basis for interactions and service use preferences of African-American woman who cared for an adult female child with a disability. Diverse African-American perspectives were observed by analyzing the discourse of 11 low-income African-American carers using a direct informant interview technique. Two distinct discourses were identified: a "secular professional" discourse and a "spiritual kin" discourse. Families using the secular professional discourse made use of formal developmental disabilities services in a manner similar to European-American heritage families. Families using the spiritual kin discourse tended to rely on natural or informal supports rather than the developmental disabilities service system, as the system did not exemplify the values that these families profess. It was observed that the discourses result in two distinct worldviews, and two distinct ways of accessing the developmental disabilities system. Recommendations are proffered for changes within the developmental disabilities service system to accommodate diverse cultures and families. A key recommendation involves modifying the system to accommodate worldviews of disability other than the currently used scientific biomedical view. [source] Synthesis and chiroptical properties of optically active poly(ethynylcarbazole) derivatives: Substituent effect on the helix formationJOURNAL OF POLYMER SCIENCE (IN TWO SECTIONS), Issue 19 2007Fumio Sanda Abstract Novel chiral acetylene monomers containing carbazole, 2-ethynyl-9-[(S)-2-methylbutoxycarbonyl]carbazole (1), 3-ethynyl-9-[(S)-2-methylbutoxycarbonyl]carbazole (2), 2-ethynyl-9-[(S)-2-methylbutyl]carbazole (3), and 2-ethynyl-9-[(S)-4-methylhexyl]carbazole (4) were synthesized and polymerized with [(nbd)RhCl]2Et3N. The corresponding polyacetylenes with number-average molecular weights ranging from 68,700 to 310,000 were obtained in good yields. Poly(1) exhibited a large specific rotation and an intense Cotton effect in toluene, indicating that it formed a helix with predominantly one-handed screw sense, while the other three polymers showed no evidence for taking a helical structure. Poly(1) largely decreased the CD intensity upon heating from ,10 to 60 °C. Poly(1) showed a Cotton effect in film state in a manner similar to solution state. No chiral amplification was observed in the copolymerization of 1 with achiral 2-ethynyl-9- tert -butoxycarbonylcarbazole (5). © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 45: 4450,4458, 2007 [source] Impaired decision making following 49 h of sleep deprivationJOURNAL OF SLEEP RESEARCH, Issue 1 2006WILLIAM D. S. KILLGORE Summary Sleep deprivation reduces regional cerebral metabolism within the prefrontal cortex, the brain region most responsible for higher-order cognitive processes, including judgment and decision making. Accordingly, we hypothesized that two nights of sleep loss would impair decision making quality and lead to increased risk-taking behavior on the Iowa Gambling Task (IGT), which mimics real-world decision making under conditions of uncertainty. Thirty-four healthy participants completed the IGT at rested baseline and again following 49.5 h of sleep deprivation. At baseline, volunteers performed in a manner similar to that seen in most samples of healthy normal individuals, rapidly learning to avoid high-risk decks and selecting more frequently from advantageous low-risk decks as the game progressed. After sleep loss, however, volunteers showed a strikingly different pattern of performance. Relative to rested baseline, sleep-deprived individuals tended to choose more frequently from risky decks as the game progressed, a pattern similar to, though less severe than, previously published reports of patients with lesions to the ventromedial prefrontal cortex. Although risky decision making was not related to participant age when tested at rested baseline, age was negatively correlated with advantageous decision making on the IGT, when tested following sleep deprivation (i.e. older subjects made more risky choices). These findings suggest that cognitive functions known to be mediated by the ventromedial prefrontal cortex, including decision making under conditions of uncertainty, may be particularly vulnerable to sleep loss and that this vulnerability may become more pronounced with increased age. [source] | ||||||||||||||||||||||||||||