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M M (m + m)
Selected AbstractsElectroanalysis of Norepinephrine at Bare Gold Electrode Pure and Modified with Gold Nanoparticles and S-Functionalized Self-Assembled Layers in Aqueous SolutionELECTROANALYSIS, Issue 13 2009Teresa, uczak Abstract Gold nanoparticles (Au-NPs), cystamine (CA) and 3,3,-dithiodipropionic acid (DTDPA) modified gold bare electrodes were applied in voltammetric sensors for simultaneous detection of norepinephrine (NEP), ascorbic (AA) and uric (UA) acids. A linear relationship between norepinephrine concentration and current response was obtained in the range of 0.1,,M to 600,,M M with the detection limit ,0.091,,M for the electrodes modified at 2D template and in the range of 0.1,,M to 700,,M M with the detection limit ,0.087,,M for the electrodes modified at 3D template The results have shown that using modified electrodes it is possible to perform electrochemical analysis of norepinephrine without interference of ascorbic and uric acids, whose presence is the major limitation in norepinephrine determination at a bare gold electrode. The modified SAMs electrodes show good selectivity, sensitivity, reproducibility and high stability. [source] RNA-dependent RNA polymerase activity encoded by GB virus-B non-structural protein 5BJOURNAL OF VIRAL HEPATITIS, Issue 5 2000Zhong Phylogenetic analysis and polyprotein organization comparison have shown that GB virus-B (GBV-B) is closely related to hepatitis C virus (HCV). In this study, the coding region for GBV-B non-structural protein 5B (NS5B) was isolated by reverse transcription,polymerase chain reaction (RT,PCR) from pooled serum of GBV-B-infected tamarins. Expression of soluble GBV-B NS5B protein in Escherichia coli was achieved by removal of a 19-amino acid hydrophobic domain at the C-terminus of the protein. The truncated GBV-B NS5B (NS5B,CT19) was purified to homogeneity and shown to possess an RNA-dependent RNA polymerase (RdRp) activity in both gel-based and scintillation proximity assays. NS5B,CT19 required the divalent cation Mn2+ for enzymatic activity, at an optimal concentration of 15 m M. Interestingly, Mg2+, at concentrations up to 20 m M, did not support the GBV-B NS5B activity. This differs from HCV NS5B where both Mn2+ and Mg2+ can support RdRp activity. Zn2+ was found to inhibit the activity of GBV-B NS5B, with a 50% inhibitory concentration (IC50) of 5,10 ,M. Higher concentrations of monovalent salts (NaCl or KCl > 100 m M) and glycerol (> 3%) were also inhibitory. NS5B,CT19 was able to bind to RNA homopolymers, but utilized most efficiently poly(C), the one with the lowest binding affinity for RNA synthesis. Mutational analysis of GBV-B NS5B demonstrated the importance of several conserved sequence motifs for enzymatic activity. Based on sequence homology (, 37% identity and 52% similarity) between GBV-B and HCV NS5B proteins, the active GBV-B RdRp provides a good surrogate assay system for HCV polymerase studies. [source] SERCA function declines with age in adrenergic nerves from the superior cervical ganglionAUTONOMIC & AUTACOID PHARMACOLOGY, Issue 5-6 2000W. J. Pottorf 1 Intracellular calcium is a universal second messenger integrating numerous cellular pathways. An age-related breakdown in the mechanisms controlling [Ca2+]i homeostasis could contribute to neuronal degeneration. One component of neuronal calcium regulation believed to decline with age is the function of sarco/endoplasmic reticulum calcium ATPase (SERCA) pumps. 2 Therefore we investigated the impact of age on the capacity of SERCA pumps to control high (68 m M) [K+]-evoked [Ca2+]i -transients in acutely dissociated superior cervical ganglion (SCG) cells from 6- and 20-month-old Fisher-344 rats. Calcium transients were measured by fura-2 microfluorometry in the presence of vanadate (0.1 ,M) to selectively block plasma membrane calcium ATPase (PMCA) pumps, dinitrophenol (100 ,M) to block mitochondrial calcium uptake and extracellular sodium replaced with tetraethylammonium to block Na+/Ca2+ -exchanger, thus forcing the neuronal cells to rely on SERCA uptake to control [Ca2+]i homeostasis. 3 In the presence of these calcium buffering blockers, the rate of recovery of [Ca2+]i was significantly slower and time to recover to approximately 90% of resting [Ca2+]i was significantly greater in SCG cells from old (20 months) compared with young (6 months) animals. 4 This age-related change in the recovery phase of [K+]-evoked [Ca2+]i -transients could not be explained by differences in the sensitivity of SCG cells to the calcium buffering blockers, as no age-related difference in basal [Ca2+]i was observed. 5 These studies illustrate that when rat SCG cells are forced to rely on SERCAs to buffer [K+]-evoked [Ca2+]i -transients, an age-related decline in SERCA function is revealed. Such age-related declines in calcium regulation coupled with neuronal sensitivity to calcium overload underscore the importance of understanding the components of [Ca2+]i homeostasis and the functional compensation that may occur with advancing age. [source] Anti-inflammatory properties of heat shock protein 70 and butyrate on Salmonella -induced interleukin-8 secretion in enterocyte-like Caco-2 cellsCLINICAL & EXPERIMENTAL IMMUNOLOGY, Issue 1 2005J. J. Malago Summary Intestinal epithelial cells secrete the chemokine interleukin (IL)-8 in the course of inflammation. Because heat shock proteins (Hsps) and butyrate confer protection to enterocytes, we investigated whether they modulate Salmonella enterica serovar Enteritidis (S. serovar Enteritidis)-induced secretion of IL-8 in enterocyte-like Caco-2 cells. Caco-2 cells incubated with or without butyrate (0,20 m M, 48 h) were infected with S. serovar Enteritidis after (1 h at 42°C, 6 h at 37°C) or without prior heat shock (37°C). Levels of Hsp70 production and IL-8 secretion were analysed using immunostaining of Western blots and enzyme-linked immunosorbent assay (ELISA), respectively. The cells secreted IL-8 in response to S. serovar Enteritidis and produced Hsp70 after heat shock or incubation with butyrate. The IL-8 secretion was inhibited by heat shock and butyrate concentrations as low as 0·2 m M for crypt-like and 1 m M for villous-like cells. In a dose-dependent manner, higher butyrate concentrations enhanced IL-8 secretion to maximal levels followed by a gradual but stable decline. This decline was associated with increasing production of Hsp70 and was more vivid in crypt-like cells. In addition, the higher concentrations abolished the heat shock inhibitory effect. Instead, they promoted the IL-8 production in heat-shocked cells even in the absence of S. serovar Enteritidis. We conclude that heat shock and low concentrations of butyrate inhibit IL-8 production by Caco-2 cells exposed to S. serovar Enteritidis. Higher butyrate concentrations stimulate the chemokine production and override the inhibitory effect of the heat shock. The IL-8 down-regulation could in part be mediated via production of Hsp70. [source] Reach-scale channel geometry of a mountain riverEARTH SURFACE PROCESSES AND LANDFORMS, Issue 8 2004Ellen Wohl Abstract Mountain rivers can be subject to strong constraints imposed by changes in gradient and grain size supplied by processes such as glaciation and rockfall. Nonetheless, adjustments in the channel geometry and hydraulics of mountain rivers at the reach scale can produce discernible patterns analogous to those in fully alluvial rivers. Mountain rivers can differ in that imposed reach-scale gradient is an especially important control on reach-scale channel characteristics, as indicated by examination of North St Vrain Creek in Colorado. North St Vrain Creek drains 250 km2 of the Rocky Mountains. We used 25 study reaches within the basin to examine controls on reach-scale channel geometry. Variables measured included channel geometry, large woody debris, grain size, and mean velocity. Drainage area at the study reaches ranged from 2·2 to 245 km2, and gradient from 0·013 to 0·147 m m,1. We examined correlations among (1) potential reach-scale response variables describing channel bankfull dimension and shape, hydraulics, bedform wavelength and amplitude, grain size, ,ow resistance, standard deviation of hydraulic radius, and volume of large woody debris, and (2) potential control variables that change progressively downstream (drainage area, discharge) or that are likely to re,ect a reach-speci,c control (bed gradient). We tested the hypothesis that response variables correlate most strongly with local bed gradient because of the segmented nature of mountain channels. Results from simple linear regression analyses indicate that most response variables correlate best with gradient, although channel width and width/depth ratio correlate best with discharge. Multiple regression analyses using Mallow's Cp selection criterion and log-transformation of all variables produced similar results in that most response variables correlate strongly with gradient. These results suggest that the hypothesis is partially supported: channel bed gradient is likely to be a good predictor for many reach-scale response variables along mountain rivers, but discharge is also an important predictor for some response variables. Copyright © 2004 John Wiley & Sons, Ltd. [source] Organic litter: dominance over stones as a source of interrill flow roughness on low-gradient desert slopes at Fowlers Gap, arid western NSW, AustraliaEARTH SURFACE PROCESSES AND LANDFORMS, Issue 1 2003David Dunkerley Abstract Thirty-six runoff plot experiments provide data on flow depths, speeds, and Darcy,Weisbach friction coefficients (f) on bare soil surfaces, and surfaces to which were added sufficient extra plant litter or surface stones to provide projected cover of 5, 10 and 20 per cent. Precision flow depth data were derived with a computer-controlled gantry and needle gauge for two different discharges for each plot treatment. Taking a fixed flow intensity (Reynolds number, Re = 150) for purposes of comparison shows means of f = 17·7 for bare soil surfaces, f = 11·4 for added stone treatments, and f = 23·8 for added litter treatments. Many individual values of f for stone treatments are lower than for the bare soil surface, but all litter treatments show increases in fcompared to bare soil. The lowering of f in stone treatments relates to the submerged volume that the stones occupied, and the associated concentration of flow onto a smaller part of the plot surface. This leads to locally higher flow intensities and lower frictional drag along threads of flow that the obstacles create. Litter causes higher frictional drag because the particles are smaller, and, for the same cover fraction, are 100 times more numerous and provide 20 times the edge or perimeter length. Along these edges, which in total exceed 2·5 m g,1 (equivalent to 500 m m,2 for a loading of 2 t ha,1), surface tension draws up water from between the litter particles. This reduces flow depth there, and as a consequence of the lower flow intensity, frictional drag rises. Furthermore, no clear passage remains for the establishment of flow threads. These findings apply to shallow interrill flows in which litter is largely immobile. The key new result from these experiments is that under these conditions, a 20 per cent cover of organic litter can generate interrill frictional retardation that exceeds by nearly 41 per cent that of a bare soil surface, and twice that contributed by the same cover fraction of surface stones. Even greater dominance by litter can be anticipated at the many dryland sites where litter covers exceed those tested here. Copyright © 2002 John Wiley & Sons, Ltd. [source] Reduced operant ethanol self-administration and in vivo mesolimbic dopamine responses to ethanol inPKC,-deficient miceEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 11 2000M. Foster Olive Abstract There is increasing evidence that individual protein kinase C (PKC) isozymes mediate specific effects of ethanol on the nervous system. In addition, multiple lines of evidence suggest that the mesoaccumbens dopamine reward system is critically involved in the rewarding and reinforcing effects of ethanol. Yet little is known about the role of individual PKC isozymes in ethanol reinforcement processes or in regulation of mesolimbic systems. In this study, we report that mice lacking the epsilon isoform of PKC (PKC,) show reduced operant ethanol self-administration and an absence of ethanol-induced increase in extracellular dopamine levels in the nucleus accumbens. PKC, null mice exhibited a 53% decrease in alcohol-reinforced operant responses under basal conditions, as well as following ethanol deprivation. Behavioural analysis revealed that while both genotypes had the same number of drinking bouts following deprivation, PKC, null mice demonstrated a 61% reduction in number of ethanol reinforcers per bout and a 57% reduction in ethanol-reinforced response rate. In vivo microdialysis experiments showed that, in contrast to wild-type mice, PKC, null mice exhibited no change in extracellular levels of dopamine in the nucleus accumbens following acute administration of ethanol (1 and 2 g/kg i.p.), while mesolimbic dopamine responses to cocaine (20 mg/kg i.p.) or high potassium (100 m m) in these mice were comparable with that of wild-types. These data provide further evidence that increases in extracellular mesolimbic dopamine levels contribute to the reinforcing effects of ethanol, and indicate that pharmacological agents inhibiting PKC, may be useful in the treatment of alcohol dependence. [source] | ||||||||||