Site-directed mutations of arginine 65 at the periphery of the active site cleft of yeast 3-phosphoglycerate kinase enhance the catalytic activity and eliminate anion-dependent activation

The function of arginine 65, a conserved residue located atthe periphery of the active site cleft in yeast 3-phospho-glyceratekinase (PGK), has been investigated by site-directed mutagenesis.Mutant enzymes with glutamine, serine and alanine at position65 all have very similar kinetic properties. The maximum velocities,determined in the absence of sulfate anion, are - 100% higherthan the V_max of wild-type PGK. The K_m values are increased2- to 3-fold for ATP and 5- to 6-fold for 3-phosphoglycerate(3PG). These results demonstrate that arginine 65 is not essentialfor catalysis. In contrast to wild-type enzyme, the mutantsare not activated by sulfate ions. In addition, steady-statekinetic experiments indicate that the mutants are no longeractivated by high concentrations of either 3PG or ATP. The dissociationconstants for anions were determined by spectral titrationsof the R65Q mutant labeled with a chromophoric probe. The K_dfor 3PG is increased 6-fold, as compared to wild-type PGK, whereasthe K_d for ATP is essentially unchanged. The KA for sulfateis decreased < 2-fold. The suppression of substrate- andsulfate-dependent activation suggests that arginine 65 participatesin the regulatory mechanism responsible for activation of theenzyme.     

Characterization of copper voids in damascene processes

The introduction of copper Damascene processing into integrated circuits has brought about a host of new defectivity issues, especially those related to pitting and voiding. These defects must be understood and eliminated to achieve competitive manufacturing yields and assure device reliability. This paper reviews various defect inspection methodologies that are useful for characterizing copper voids and their electrical effects.     

A brief history of vascular access for hemodialysis: an unfinished story

In the recent literature an inverse relationship between iron status and serum aluminum levels has repeatedly been reported in dialysis patients. To check whether this observation is, at least in part, due to an interference of iron with the protein binding of aluminum, we studied the effect of the latter element on both the number of free binding sites on transferrin (Tf) and on the affinity of the protein for aluminum. For the purpose of this, a recently developed HPLC-ETAAS hybrid method was used, allowing protein-binding studies at clinical relevant metal concentrations and under contamination-free conditions. After we incubated apo-Tf with iron and aluminum which were added in amounts equivalent to the calculated number of metal-binding sites on the protein (i.e., 2 mol metal/mol Tf), we found that Tf can be saturated for 100% with iron. However, for aluminum only a 23% aluminum-Tf saturation was observed. In Tf solutions with iron saturations ranging between 0 and 60% as well as in the serum of 15 subjects with iron-Tf saturations varying between 12 and 48%, a significant (p < 0.001) negative correlation between the degree of iron-Tf saturation and the percentage of aluminum (added in amounts equivalent to the number of the remaining binding sites on Tf) bound to Tf was noted (y = -0.26x + 24.5, r = -0.87 in serum). It is concluded that the iron-Tf saturation influences the Tf binding of aluminum not only by occupying binding sites otherwise available for aluminum, but also by lowering the affinity of Tf for aluminum. The effects of iron on serum aluminum levels and bone aluminum deposition are discussed.     

Altered receptive fields and sensory modalities of rat VPL thalamic neurons during spinal strychnine-induced allodynia

Altered receptive fields and sensory modalities of rat VPL thalamic neurons during spinal strychnine-induced allodynia. J. Neurophysiol. 78: 2296-2308, 1997. Allodynia is an unpleasant sequela of neural injury or neuropathy that is characterized by the inappropriate perception of light tactile stimuli as pain. This condition may be modeled experimentally in animals by the intrathecal (i.t.) administration of strychnine, a glycine receptor antagonist. Thus after i.t. strychnine, otherwise innocuous tactile stimuli evoke behavioral and autonomic responses that normally are elicited only by noxious stimuli. The current study was undertaken to determine how i.t. strychnine alters the spinal processing of somatosensory input by examining the responses of neurons in the ventroposterolateral thalamic nucleus. Extracellular, single-unit recordings were conducted in the lateral thalamus of 19 urethan-anaesthetized, male, Wistar rats (342 +/- 44 g; mean +/- SD). Receptive fields and responses to noxious and innocuous cutaneous stimuli were determined for 19 units (1 per animal) before and immediately after i.t. strychnine (40 microgram). Eighteen of the animals developed allodynia as evidenced by the ability of otherwise innocuous brush or air jet stimuli to evoke cardiovascular and/or motor reflexes. All (3) of the nociceptive-specific units became responsive to brush stimulation after i.t. strychnine, and one became sensitive to brushing over an expanded receptive field. Expansion of the receptive field, as determined by brush stimulation, also was exhibited by all of the low-threshold mechanoreceptive units (14) and wide dynamic range units (2) after i.t. strychnine. The use of air jet stimuli at fixed cutaneous sites also provided evidence of receptive field expansion, because significant unit responses to air jet developed at 13 cutaneous sites (on 7 animals) where an identical stimulus was ineffective in evoking a unit response before i.t. strychnine. However, the magnitude of the unit response to cutaneous air jet stimulation was not changed at sites that already had been sensitive to this stimulus before i.t. strychnine. The onset of allodynia corresponded with the onset of the altered unit responses (i.e., lowered threshold/receptive field expansion) for the majority of animals (9), but the altered unit response either terminated concurrently with symptoms of allodynia (6) or, more frequently, outlasted the symptoms of allodynia (10) as the effects of strychnine declined. The present results demonstrate that the direct, receptor-mediated actions of strychnine on the spinal processing of sensory information are reflected by changes in the receptive fields and response properties of nociceptive and nonnociceptive thalamic neurons. These changes are consistent with the involvement of thalamocortical mechanisms in the expression of strychnine-induced allodynia and, moreover, suggest that i.t. strychnine also produces changes in innocuous tactile sensation.     

Synthesis and Dielectric Characterization of Potassium Niobate Tantalate Ceramics

The preparation and dielectric properties of potassium niobate tantalate (KTN) have been investigated with the aim of exploring the material's potential for ferroelectric tunable applications. The samples were prepared both by conventional sintering in air and by uniaxial hot pressing. A relative average density greater than 92% was obtained with both methods. An inhomogeneous Nb/Ta distribution was found in the samples prepared by both methods, but the inhomogeneity extent was lower in the hot-pressed samples. While both sintering processes resulted in ceramics of lower transition temperatures in comparison with the reported results on single crystals, a relative up shift of the temperature ( T _max) at which the dielectric constant is maximum was found for hot-pressed samples compared with that of samples sintered in air. All the samples exhibited strong frequency dispersion in dielectric properties. The effect of DC bias at room temperature was measured and modeled using the Landau–Devonshire model. It was found that the nonlinear coefficient β of KTN, which is important for tunable applications, is comparable with the value reported on KNbO₃ and SrTiO₃ single crystals. A dielectric tunability of 16% and 42% at room temperature was demonstrated under 20 kV/cm for the sintered and hot-pressed ceramics, respectively. The loss tangent, low at megahertz frequencies, was augmented to 9%–17% at low gigahertz frequencies. This is believed to be related to the frequency dispersion observed in the ceramics because of charged defects.     

Prejudice and Stereotype Maintenance Processes: Attention, Attribution, and Individuation.

Three experiments examined the relationship between prejudice and processing of stereotypic information. Higher levels of prejudice were associated with greater attention to and more thorough encoding of stereotype-inconsistent than stereotype-consistent behaviors but only when processing capacity was plentiful (Experiments 1 and 3). High-prejudice participants attributed consistent behaviors to internal factors and inconsistent behaviors to external forces (Experiment 2). Together, these results suggest that high-prejudice people attend carefully to inconsistent behaviors to explain them away but only if they have sufficient resources to do so. Results also showed that low-prejudice but not high-prejudice participants formed individuated impressions by integrating the implications of the target's behaviors (i.e., individuating). High levels of prejudice appear to be associated with biased encoding and judgment processes that may serve to maintain stereotypes. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Local-scale analysis of carbon mitigation strategies: Tompkins County,New York,USA

The costs and potential for several carbon mitigation options were analyzed for Tompkins County, NY, within several categories: terrestrial carbon sequestration, local power generation, transportation, and energy end-use efficiency. The total county emissions are about 340 Gg C/year, with current biomass sequestration rates of about 121 Gg C/year. The potential for mitigation with the options examined, assuming full market penetration, amounts to at least 234 Gg C/year (69%), with 100 Gg C/year (29%) at no net cost to the consumer. Effective carbon mitigation strategies for this county based on costs per mg carbon and maximum potential include reforestation of abandoned agricultural lands for terrestrial carbon sequestration, biomass production for residential heating and co-firing in coal power plants, changes in personal behavior related to transportation (e.g., carpooling or using public transportation), installation of numerous residential energy-efficient products and development of local wind power. The principal barriers to the implementation of these approaches are discussed and policies for overcoming these barriers are analyzed.     

Long-term trends and opportunities for managing regional water supply and wastewater greenhouse gas emissions

Greenhouse gas emissions are likely to rise faster than growth in population and more than double for water supply and wastewater services over the next 50 years in South East Queensland (SEQ), Australia. New sources of water supply such as rainwater tanks, recycled water, and desalination currently have greater energy intensity than traditional sources. In addition, direct greenhouse gas emissions from reservoirs and wastewater treatment and handling have potentially the same magnitude as emissions from the use of energy. Centralized and decentralized water supply and wastewater systems are considered for a scenario based upon a government water supply strategy for the next 50 years. Many sources of data have large uncertainties which are estimated following the IPCC Good Practice Guidelines. Important sources of emissions with large uncertainties such as rainwater tanks and direct emissions were identified for further research and potential mitigation of greenhouse gas emissions.     

End-to-End Rate Adaptation to Support Heterogeneous Services for Infrastructure-Based Vehicular Networks

Vehicle to Infrastructure(V2I) communications aim to provide mobile users on the road low-cost Internet and driver safety services.However, to meet Quality of Service(QoS) requirements of various applications and efficiently utilize limited wireless channel resourc-es, the transport layer protocol has to perform effective rate control in low channel quality and frequent changing topology communica-tion environment. In this paper, we propose a novel rate-control scheme in infrastructure based vehicular networks that avoids conges-tion and starvation and promotes fairness in end-to-end V2I communications. In vehicular networks, a bottleneck roadside unit(RSU)keeps track of its buffer size, aggregate incoming rate, and link throughput, and appropriately allocates bandwidth to traversing flows.With feedback information from the RSU, source nodes dynamically adjust their sending rates to avoid buffer overflow or starvation atthe bottleneck RSU. Simulation results show that the proposed scheme can reduce not only packet losses owing to buffer overflow butalso buffer starvation time, which improves the utilization efficiency of wireless channel resource.     

Explicit Rate Based Transmission Control Scheme in Vehicle-to-lnfrastructure Communication Networks

In Vehicle-to-Infrastructure （V2I） communication networks, mobile users are able to access Internet services, such as video stre- aming, digital map downloading, database acc- ess, online gaming, and even safety services like accident alarm, traffic condition broadcast, etc., through fixed roadside units. However, the dynamics of communication environment and frequent changing topology critically challe- nge the design of an efficient transport layer protocol, which makes it difficult to guarantee diverse Quality of Service （QoS） requirements for various applications. In this paper, we pre- sent a novel transport layer scheme in infra- structure based vehicular networks, and aim to resolve some challenging issues such as sou- rce transfer rate adjustment, congestion avoid- ance, and fairness. By precisely detecting pac- ket losses and identifying various causes of th- ese losses （for example, link disconnection, cha- nnel error, packet collision, buffer overflow）, the proposed scheme adopts different reacting mechanisms to deal with each of the losses. Moreover, it timely monitors the buffer size of the bottleneck Road-Side Unit （RSU）, and dyn- amically makes transfer rate feedbacks to sou- rce nodes to avoid buffer overflow or vacancy. Finally, analysis and simulation results show that the proposed scheme not only successfully reduces packet losses because of buffer over- flow and link disconnection but also improves the utilization efficiency of channel resource.