Membrane transport mechanisms probed by capacitance measurements with megahertz voltage clamp.

We have used capacitance measurements with a 1-microsecond voltage clamp technique to probe electrogenic ion-transporter interactions in giant excised membrane patches. The hydrophobic ion dipicrylamine was used to test model predictions for a simple charge-moving reaction. The voltage and frequency dependencies of the apparent dipicrylamine-induced capacitance, monitored by 1-mV sinusoidal perturbations, correspond to single charges moving across 76% of the membrane field at a rate of 9500 s-1 at 0 mV. For the cardiac Na,K pump, the combined presence of cytoplasmic ATP and sodium induces an increase of apparent membrane capacitance which requires the presence of extracellular sodium. The dependencies of capacitance changes on frequency, voltage, ATP, and sodium verify that phosphorylation enables a slow, 300- to 900-s-1, pump transition (the E1-E2 conformational change), which in turn enables fast, electrogenic, extracellular sodium binding reactions. For the GAT1 (gamma-aminobutyric acid,Na,Cl) cotransporter, expressed in Xenopus oocyte membrane, we find that chloride binding from the cytoplasmic side, and probably sodium binding from the extracellular side, results in a decrease of membrane capacitance monitored with 1- to 50-kHz perturbation frequencies. Evidently, ion binding by the GAT1 transporter suppresses an intrinsic fast charge movement which may originate from a mobility of charged residues of the transporter binding sites. The results demonstrate that fast capacitance measurements can provide new insight into electrogenic processes closely associated with ion binding by membrane transporters.

A chimeric light-regulated amino acid transport system allows the isolation of blue light regulator (blr) mutants of Neurospora crassa.

We have developed a system for the isolation of Neurospora crassa mutants that shows altered responses to blue light. To this end we have used the light-regulated promoter of the albino-3 gene fused to the neutral amino acid permease gene mtr. The product of the mtr gene is required for the uptake of neutral aliphatic and aromatic amino acids, as well as toxic analogs such as p-flurophenylalanine or 4-methyltryptophan. mtr trp-2-carrying cells were transformed with the al-3 promoter-mtr wild-type gene (al-3p-mtr+) to obtain a strain with a light-regulated tryptophan uptake. This strain is sensitive to p-fluorophenylalanine when grown under illumination and resistant when grown in the dark. UV mutagenesis of the al-3p-mtr(+)-carrying strain allowed us to isolate two mutant strains, BLR-1 and BLR-2 (blue light regulator), that are light-resistant to p-fluorophenylalanine and have lost the ability to grow on tryptophan. These two strains have a pale-orange phenotype and show down-regulation of all the photoregulated genes tested (al-3, al-1, con-8, and con-10). Mutations in the BLR strains are not allelic with white collar 1 or white collar 2, regulatory genes that are also involved in the response to blue light.

Phospholipase D is present on Golgi-enriched membranes and its activation by ADP ribosylation factor is sensitive to brefeldin A.

ADP ribosylation factor (ARF) is a small guanosine triphosphate (GTP)-binding protein that regulates the binding of coat proteins to membranes and is required for several stages of vesicular transport. ARF also stimulates phospholipase D (PLD) activity, which can alter the lipid content of membranes by conversion of phospholipids into phosphatidic acid. Abundant PLD activity was found in Golgi-enriched membranes from several cell lines. Golgi PLD activity was greatly stimulated by ARF and GTP analogs and this stimulation could be inhibited by brefeldin A (BFA), a drug that blocks binding of ARF to Golgi membranes. Furthermore, in Golgi membranes from BFA-resistant PtK1 cells, basal PLD activity was high and not stimulated by exogenous ARF or GTP analogs. Thus, ARF activates PLD on the Golgi complex, suggesting a possible link between transport events and the underlying architecture of the lipid bilayer.

Differential subcellular mRNA targeting: deletion of a single nucleotide prevents the transport to axons but not to dendrites of rat hypothalamic magnocellular neurons.

It has previously been shown that mRNA encoding the arginine vasopressin (AVP) precursor is targeted to axons of rat magnocellular neurons of the hypothalamo-neurohypophyseal tract. In the homozygous Brattle-boro rat, which has a G nucleotide deletion in the coding region of the AVP gene, no such targeting is observed although the gene is transcribed. RNase protection and heteroduplex analyses demonstrate that, in heterozygous animals, which express both alleles of the AVP gene, the wild-type but not the mutant transcript is subject to axonal compartmentation. In contrast, wild-type and mutant AVP mRNAs are present in dendrites. These data suggest the existence of different mechanisms for mRNA targeting to the two subcellular compartments. Axonal mRNA localization appears to take place after protein synthesis; the mutant transcript is not available for axonal targeting because it lacks a stop codon preventing its release from ribosomes. Dendritic compartmentation, on the other hand, is likely to precede translation and, thus, would be unable to discriminate between the two mRNAs.

Emergence of half-metallicity in suspended NiO chains: Ab initio electronic structure and quantum transport calculations

Contrary to the antiferromagnetic and insulating character of bulk NiO, one-dimensional chains of this material can become half metallic due to the lower coordination of their atoms. Here we present ab initio electronic structure and quantum transport calculations of ideal infinitely long NiO chains and of more realistic short ones suspended between Ni electrodes. While infinite chains are insulating, short suspended chains are half-metallic minority-spin conductors that displays very large magnetoresistance and a spin-valve behavior controlled by a single atom.

Impacts on particles and ozone by transport processes recorded at urban and high-altitude monitoring stations

In order to evaluate the influence of particle transport episodes on particle number concentration temporal trends at both urban and high-altitude (Aitana peak-1558 m a.s.l.) stations, a simultaneous sampling campaign from October 2011 to September 2012 was performed. The monitoring stations are located in southeastern Spain, close to the Mediterranean coast. The annual average value of particle concentration obtained in the larger accumulation mode (size range 0.25–1 μm) at the mountain site, 55.0 ± 3.0 cm− 3, was practically half that of the value obtained at the urban station (112.0 ± 4.0 cm− 3). The largest difference between both stations was recorded during December 2011 and January 2012, when particles at the mountain station registered the lowest values. It was observed that during urban stagnant episodes, particle transport from urban sites to the mountain station could take place under specific atmospheric conditions. During these transports, the major particle transfer is produced in the 0.5–2 μm size range. The minimum difference between stations was recorded in summer, particularly in July 2012, which is most likely due to several particle transport events that affected only the mountain station. The particle concentration in the coarse mode was very similar at both monitoring sites, with the biggest difference being recorded during the summer months, 0.4 ± 0.1 cm− 3 at the urban site and 0.9 ± 0.1 cm− 3 at the Aitana peak in August 2012. Saharan dust outbreaks were the main factor responsible for these values during summer time. The regional station was affected more by these outbreaks, recording values of > 4.0 cm− 3, than the urban site. This long-range particle transport from the Sahara desert also had an effect upon O3 levels measured at the mountain station. During periods affected by Saharan dust outbreaks, ozone levels underwent a significant decrease (3–17%) with respect to its mean value.

ROCK OUTCROPS IN THE CANADIAN SHIELD: AN INVESTIGATION OF CONTAMINANT TRANSPORT FROM SURFACE SOURCES IN FRACTURED ROCK AQUIFERS

Flow, recharge and transport dynamics in fractured rock aquifers with low lying rock outcrops is a largely unexplored area of study in hydrogeology. The purpose of this thesis is to examine these topics in an agricultural area in Eastern Ontario. The study consists of a regional scale groundwater quality study, an infiltration experiment that considers bacteria transport from the ground surface to a well, and a numerical modelling study that tests the parameters that affect surface infiltration of a tracer from a rock outcrop to a deeper horizontal fracture. In the water quality study, approximately 65% of the samples contained total coliform, 16% contained E. coli, and 1% contained nitrate-N at greater than 5 mg/L. Occurrence of E. coli increased when considering seasonality, where wells were drilled on rock outcrops, and for shallow well intervals. Nitrate-N did not occur above the Guidelines for Canadian Drinking Water Quality (Health Canada, 2012) of 10 mg/L. Rapid arrival times were observed in the infiltration study for both the microspheres (30 minutes) and a dye tracer (45 minutes) in a well approximately 6.0 m in horizontal and 2.8 m in vertical distance from the tracer source. Transport velocities were approximately 38.9 m/day for the dye tracer and 115.2 m/day for the colloidal tracer. Results of the model runs indicate that overburden can provide an effective protective layer to transport in fractures, that high groundwater velocities occur in larger fracture apertures and higher gradients dilute tracer concentrations, and that lower groundwater velocities occur with smaller fracture apertures and lower gradients result in elevated tracer concentrations. Lower rainfall rates, larger fracture apertures, early tracer time, larger gradients, and lower water levels maintained unsaturated conditions for longer time periods such that tracer transport was delayed until saturated conditions were attained. The overall heterogeneity of this aquifer environment creates a source water protection conundrum where the water quality is generally good, while transport can occur very quickly in proximity to rock outcrops and in areas with limited overburden.

Analyse de la distribution d'Escherichia coli, utilisée comme un marqueur microbiologique, dans un réseau de production porcin

La présence d’Escherichia coli pathogènes en élevages porcins entraine des retards de croissance et la mortalité. La transmission des E. coli pathogènes entre les élevages et l'abattoir d’un même réseau de production n'est pas bien décrite. La détection des gènes de virulence des E. coli pathogènes pourrait permettre d’identifier un marqueur de contamination dans le réseau. L’objectif de cette étude a été d’identifier un marqueur de contamination E. coli dans un réseau de production porcine défini afin de décrire certains modes de transmission des E. coli pathogènes. Pour ce faire, une région géographique comprenant 10 fermes d’engraissement, un abattoir et un réseau de transport a été sélectionnée. Trois lots de production consécutifs par ferme ont été suivis pendant 12 mois. Des échantillons environnementaux ont été prélevés à l’intérieur et à l’extérieur des fermes (3 visites d’élevage), dans la cour de l’abattoir (2 visites lors de sorties de lot) et sur le camion de transport. La détection des gènes de virulence (eltB, estA, estB, faeG, stxA, stx2A, eae, cnf, papC, iucD, tsh, fedA) dans les échantillons a été réalisée par PCR multiplexe conventionnelle. La distribution temporelle et spatiale des gènes de virulence a permis d’identifier le marqueur de contamination ETEC/F4 défini par la détection d’au moins un gène d’entérotoxine ETEC (estB, estA et eltB) en combinaison avec le gène de l’adhésine fimbriaire (faeG). La distribution des échantillons positifs ETEC/F4 qualifie la cour de l’abattoir comme un réservoir de contamination fréquenté par les transporteurs, vecteurs de contamination entre les élevages. Ceci suggère le lien microbiologique entre l’élevage, les transporteurs et l’abattoir jouant chacun un rôle dans la dissémination des microorganismes pathogènes et potentiellement zoonotiques en production porcine.

Identification and characterization of GABA, proline and quaternary ammonium compound transporters from Arabidopsis thaliana

Arabidopsis thaliana grows efficiently on GABA as the sole nitrogen source, thereby providing evidence for the existence of GABA transporters in plants. Heterologous complementation of a GABA uptake-deficient yeast mutant identified two previously known plant amino acid transporters, AAP3 and ProT2, as GABA transporters with Michaelis constants of 12.9±1.7 and 1.7±0.3 mM at pH 4, respectively. The simultaneous transport of [1-14C]GABA and [2,3-3H]proline by ProT2 as a function of pH, provided evidence that the zwitterionic state of GABA is an important parameter in substrate recognition. ProT2-mediated [1-14C]GABA transport was inhibited by proline and quaternary ammonium compounds.

Optimum Design of Electromagnets for Magnetic Levitation of Transport Systems based on the Inverse Problem Solutions

Article is devoted to design of optimum electromagnets for magnetic levitation of transport systems. The method of electromagnets design based on the inverse problem solution of electrical equipment is offered. The method differs from known by introducing a stage of minimization the target functions providing the stated levitation force and magnetic induction in a gap, and also the mass of an electromagnet. Initial values of parameters are received, using approximate formulas of the theory of electric devices and electrical equipment. The example of realization of a method is given. The received results show its high efficiency at design. It is practical to use the offered method and the computer program realizing it as a part of system of the automated design of electric equipment for transport with a magnetic levitation.

Analysis of coastal sediment transport processes from Wrightsville Beach to Fort Fisher, North Carolina /

A comprehensive engineering analysis of the coastal sediment transport processes along a 42-kilometer segment of the North Carolina shoreline from Wrightsville Beach to Fort Fisher is presented. Included in the analysis is an interpretation of the littoral processes, longshore transport, and the behavior and success of beach nourishment projects at Wrightsville Beach and Carolina Beach, North Carolina. The historical position of the MLW, MSL, and MHW contours, relative to a fixed base line, is plotted for the period between 1964 and 1975. An equivalent volumetric erosion or accretion between successive surveys is determined by multiplying the average excursion distance of the contours by a constant of proportionality. The plots of excursion distance versus time for the MLW, MSL, and MHW contours also show the time response of the beach fills. This response is described by a mathematical function. The alongshore components of wave-induced energy flux are also determined within the study area through wave refraction analysis. This information, together with the information on volumetric change, is used in a sediment budget analysis to determine the coefficient of alongshore sediment transport and the inlet trapping characteristics. (Author).

Instrumentation for measuring pavement-vehicle interaction. Volume III: Kennedy Co. operation and maintenance manual, formatter and digital tape transport. Final report.

Federal Highway Administration, Structures and Applied Mechanics Division, Washington, D.C.

Processes affecting subsurface transport of leaking underground tank fluids /

"CR 810052."

Summary report - full-scale transport controlled impact demonstration program. Final report.

Mode of access: Internet.

The performance of child restraint devices in transport airplane passenger seats. Final report.

Federal Aviation Administration, Washington, D.C.