Effect of intertidal exposure on Vibrio parahaemolyticus levels in Pacific Northwest oysters

Interest in Vibrio parahaemolyticus (Vp) increased in the United States following Vp-associated gastroenteritis outbreaks in 1997 and 1998 involving the West Coast and other areas. The present study evaluated multiple aspects of Vp ecology in the Pacific Northwest with three objectives: (i) to determine the effect of low-tide exposure on Vp levels in oysters, (ii) to determine the relationship between total and pathogenic Vp, and (iii) to examine sediments and aquatic fauna as reservoirs for pathogenic Vp. Samples were collected from intertidal reefs along Hood Canal, Wash., in August 2001. Fecal matter from marine mammals and aquatic birds as well as intestinal contents from bottom-dwelling fish were tested. Total and pathogenic Vp levels in all the samples were enumerated with colony hybridization procedures using DNA probes that targeted the thermolabile direct hemolysin (tlh) and thermostable direct hemolysin (tdh) genes, respectively. The mean Vp densities in oysters were four to eight times greater at maximum exposure than at the corresponding first exposure. While tdh-positive Vp counts were generally < or = 10 CFU/g at first exposure, counts as high as 160 CFU/g were found at maximum exposure. Vp concentrations in sediments were not significantly different from those in oysters at maximum exposure. Pathogenic (tdh positive) Vp was detected in 9 of 42 (21%) oyster samples at maximum exposure, in 5 of 19 (26%) sediment samples, but in 0 of 9 excreta samples. These results demonstrate that summer conditions permit the multiplication of Vp in oysters exposed by a receding tide.     

Real-time PCR quantification of Vibrio parahaemolyticus in oysters using an alternative matrix

This study examined the relationship between levels of total Vibrio parahaemolyticus found in oyster tissues and mantle fluid with the goal of using mantle fluid as a template matrix in a new quantitative real-time PCR assay targeting the thermolabile hemolysin (tlh) gene for the enumeration of total V. parahaemolyticus in oysters. Oysters were collected near Mobile Bay, Ala., in June, July, and September and tested immediately after collection and storage at 26 degrees C for 24 h. Initial experiments using DNA colony hybridization targeting tlh demonstrated that natural V. parahaemolyticus levels in the mantle fluid of individual oysters were strongly correlated (r = 0.85, P < 0.05) with the levels found in their tissues. When known quantities of cultured V. parahaemolyticus cells were added to real-time PCR reactions that contained mantle fluid and oyster tissue matrices separately pooled from multiple oysters, a strong linear correlation was observed between the real-time PCR cycle threshold and the log concentration of cells inoculated into each PCR reaction (mantle fluid: r = 0.98, P < 0.05; and oyster: r = 0.99, P < 0.05). However, the mantle fluid exhibited less inhibition of the PCR amplification than the homogenized oyster tissue. Analysis of natural V. parahaemolyticus populations in mantle fluids using both colony hybridization and real-time PCR demonstrated a significant (P < 0.05) but reduced correlation (r = -0.48) between the two methods. Reductions in the efficiency of the real-time PCR that resulted from low population densities of V. parahaemolyticus and PCR inhibitors present in the mantle fluid of some oysters (with significant oyster-to-oyster variation) contributed to the reduction in correlation between the methods that was observed when testing natural V. parahaemolyticus populations. The V. parahaemolyticus-specific real-time PCR assay used for this study could estimate elevated V. parahaemolyticus levels in oyster mantle fluid within 1 h from sampling time.     

Optimum design of pendulum‐type tuned mass dampers

The equations of motion are derived for a translational single degree of freedom system equipped with a ‘pendulum‐type’ tuned mass damper (TMD) under dynamic force and base acceleration excitations. The complex frequency response functions are obtained. Following response minimization procedures, the optimum parameters of the TMD under random white noise excitations are determined. The effect of the TMD in reducing the response is expressed in terms of an equivalent viscous damping. The optimum design parameters and the corresponding efficiency of the TMD under both wind and earthquake dynamic loads are presented in design charts. The effect of the structure inherent and aerodynamic damping on the optimum parameters is studied and simplified charts to account for such effect are provided. Moreover, a design chart for the over‐optimum‐damped TMDs is presented. The translational‐type TMD is treated as a special case of the pendulum‐type. Copyright © 2005 John Wiley & Sons, Ltd.     

Reliability analysis of wind response of flexibly supported tall structures

Statistics and probabilistic analyses and risk assessments can be very useful decision‐making tools when dealing with structural–geotechnical problems. Wind loads, dynamic properties of soil underneath the structure and material characteristics of the structure are important factors that affect the wind action on the structure and consequently the structural wind‐induced response. Uncertainties in the estimation of these factors as a result of human error or inherent variability are at the forefront for the use of reliability approaches to evaluate the risk of failure during the service period. In the present study, probabilistic base force analyses for tall structures are performed. The substructure approach in which the soil supporting the foundation is modeled by the foundation compliances as functions of soil shear wave velocity is used to account for the soil–structure interaction efficiently. A three‐variable probabilistic approach is used to account for the uncertainties in shear wave velocity of the soil underneath the foundation, the concrete strength and the design wind speed on the calculated response and the base forces. The second moment approximation using Taylor series expansion is used to perform the probabilistic analyses of the base cross‐section design and resistant forces of a free‐standing tower. The first‐order reliability method is used to examine the failure probability and the contribution to the total uncertainty. The results show that the dynamic response of the tower increases as soil shear wave velocity decreases. For the range of soil shear wave velocity encountered in practice, the base forces of the structure may increase by up to 20% as a result of the foundation flexibility. For the limit state considered in this study, it was found that the reliability index decreases by up to 15% and the probability of failure increases by up to one order of magnitude as a result of the soil–structure interaction effect. Copyright © 2003 John Wiley & Sons, Ltd.     

Charge pair interactions stabilizing ferredoxin-ferredoxin reductase complexes. Identification by complementary site-specific mutations

Ferredoxin reductase (Fd-reductase) supplies electrons to mitochondrial steroid hydroxylase cytochrome P450 enzymes via a [2Fe-2S] ferredoxin. Chemical labeling studies with bovine Fd-reductase have implicated Lys-243 as important in binding to bovine ferredoxin (Hamamoto, I., Kazutaka, K., Tanaka, S., and Ichikawa, Y. (1988) Biochim. Biophys. Acta 953, 207-213). We have used site-directed mutagenesis to examine the role of charged residues in this region of human Fd-reductase in ferredoxin binding. Mutant proteins were expressed in Escherichia coli and were assayed for activity by ferredoxin-mediated electron transfer to cytochrome c. Replacement of Lys-242 (homologous to Lys-243 in bovine Fd-reductase) with Gln and replacement of Arg-241 with Ser had little effect (2.7- and 3.6-fold increased Km, respectively). In contrast, mutants at positions 239 and 243 (R239S and R243Q) exhibited markedly lower affinity for ferredoxin (17.5- and 1,600-fold increased Km, respectively). Studies were also carried out with two ferredoxin charge mutants shown previously to have lowered affinity for Fd-reductase (Coghlan, V. M., and Vickery, L. E. (1991) J. Biol. Chem. 266, 18606-18612). Comparisons of the binding of ferredoxin mutants D76N and D79N to Fd-reductase mutants R239S and R243Q suggest that Arg-239 and Arg-243 of Fd-reductase each interact directly with both Asp-76 and Asp-79 of ferredoxin during formation of the complex between the two proteins. These results support the hypothesis that specific electrostatic interactions involving this region are important in stabilizing the ferredoxin-Fd-reductase complex.     

Cognitive skill and traditional trance hypnotic inductions: A within-subjects comparison.

Comparison of a traditional trance hypnotic induction and a cognitive skill induction in a within-Ss design with 40 undergraduates showed that the cognitive skill induction enhanced subjective responses to suggestions and produced significant increments in behavioral responses when it was preceded by the trance induction. The trance procedure led to greater self-reported alterations in consciousness. Findings suggest that skill induction teaches cognitive strategies that enhance responsivity to suggestions in subsequent hypnotic experiences, independent of alterations in consciousness elicited by trance induction. (5 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Effect of an impact-generated gas cloud on entrained surface boulders and spall ejecta

The purpose of this investigation is to study the effect of an impact-generated gas cloud on the size-velocity distribution of large, solid ejecta fragments that become entrained in it, with particular reference to the possible Martian origin of the SNC meteorites. The entrainment both of loose surface boulders and of the early-time, large-size, high-velocity spall component of the crater ejecta is modeled numerically. Surface boulders from as far as 40 km from the center of impact can be accelerated by the high velocity leading edge of the gas cloud to velocities in excess of Martian escape velocity (5 km/s), but are generally crushed by the acceleration. Spall fragments become entrained later and nearer the center of the gas cloud, where gas velocities are much less. High velocity spalls are decelerated by the gas and low velocity spalls are accelerated by the gas, but no spalls ejected at < 5km/s are accelerated to velocities 5km/s. An impact-generated gas cloud is thus expected to scour the pre-existing surface of loose material and to change the size-velocity distribution of spall ejecta, but does not sufficiently enhance the velocities of crater ejecta to explain the Martian origin of SNC meteorites as large rocks.     

The response of reinforced concrete chimneys to vortex shedding

Predicted vortex induced response of circular cross-section reinforced concrete structures is compared with observed full-scale behaviour. On average the predictions agree to within 5% with observations but have a large scatter attributable to the model's failure to recognize the dependence of aerodynamic parameters on free stream turbulence. The high coefficient of variation of the model is, however, not markedly different from that associated with the prediction of drag loads.     

DNA binding of some chiral metallointercalators derived from 9,10-phenanthrenediamine

A study of the interaction with calf thymus DNA is described of a novel set of chiral ternary complex cations of general form [Ru(N(4)-tet)(phdi)](2+) (where N(4)-tet is the chiral linear tetradentate R(*)R(*)-picchxn or R(*)-picchxnMe(2)). Individual equilibrium binding constants (K(B)) have been determined from spectroscopic titrations employing the hypochromism induced in the visible absorbance of the cations on interaction with the nucleic acid. These demonstrate both stereo- and enantioselectivity in the binding interactions. These K(B) data, together with induced circular dichroism and DNA thermal denaturation results, are all indicative of selective intercalation of the bidentate components of the cations into the nucleobase stack of the duplex. Supportive evidence for a secondary binding mode for the picchxn complexes is provided by the different mutagenicity profiles obtained for related cations.