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In the present study, a new vibratory conveyor (patent pending) equipped with a single electro-vibrator intended for an accurate material dosage is investigated. The possibilities of material transportation in the circum-resonant zone were investigated analytically and by simulations [1]. Furthermore, the dependencies of the transport velocity of the tested conveyor as functions of the excitation frequency were determined. Favorable excitation frequencies at transports in the main and reversal directions were found, and the high usefulness of the machine in the production lines requiring accurate material dosage was indicated. A control strategy allowing for a sudden stop of the transported material was also proposed.

In the present study, a new vibratory conveyor (patent pending) equipped with a single electro-vibrator intended for an accurate material dosage is investigated. The possibilities of material transportation in the circum-resonant zone were investigated analytically and by simulations [1]. Furthermore, the dependencies of the transport velocity of the tested conveyor as functions of the excitation frequency were determined. Favorable excitation frequencies at transports in the main and reversal directions were found, and the high usefulness of the machine in the production lines requiring accurate material dosage was indicated. A control strategy allowing for a sudden stop of the transported material was also proposed. [ABSTRACT FROM AUTHOR]

Abstract Background Much of the structure-based mechanistic understandings of the function of SLC6A neurotransmitter transporters emerged from the study of their bacterial LeuT-fold homologs. It has become evident, however, that structural differences such as the long N- and C-termini of the eukaryotic neurotransmitter transporters are involved in an expanded set of functional properties to the eukaryotic transporters. These functional properties are not shared by the bacterial homologs, which lack the structural elements that appeared later in evolution. However, mechanistic insights into some of the measured functional properties of the eukaryotic transporters that have been suggested to involve these structural elements are sparse or merely descriptive. Results To learn how the structural elements added in evolution enable mechanisms of the eukaryotic transporters in ways not shared with their bacterial LeuT-like homologs, we focused on the human dopamine transporter (hDAT) as a prototype. We present the results of a study employing large-scale molecular dynamics simulations and comparative Markov state model analysis of experimentally determined properties of the wild-type and mutant hDAT constructs. These offer a quantitative outline of mechanisms in which a rich spectrum of interactions of the hDAT N-terminus and C-terminus contribute to the regulation of transporter function (e.g., by phosphorylation) and/or to entirely new phenotypes (e.g., reverse uptake (efflux)) that were added in evolution. Conclusions The findings are consistent with the proposal that the size of eukaryotic neurotransmitter transporter termini increased during evolution to enable more functions (e.g., efflux) not shared with the bacterial homologs. The mechanistic explanations for the experimental findings about the modulation of function in DAT, the serotonin transporter, and other eukaryotic transporters reveal separate roles for the distal and proximal segments of the much larger N-terminus in eukaryotic transporters compared to the bacterial ones. The involvement of the proximal and distal segments — such as the role of the proximal segment in sustaining transport in phosphatidylinositol 4,5-bisphosphate-depleted membranes and of the distal segment in modulating efflux — may represent an evolutionary adaptation required for the function of eukaryotic transporters expressed in various cell types of the same organism that differ in the lipid composition and protein complement of their membrane environment.

Our aim was to identify and quantify the major in vivo pathways of lipoprotein cholesteryl ester transport in humans. Normal (n = 7), bile fistula (n = 5), and familial hypercholesterolemia (FH; n = 1) subjects were studied. Each received isotopic free cholesterol in HDL, LDL, or particulate form, along with another isotope of free or esterified cholesterol or mevalonic acid. VLDL, intermediate density lipoprotein (IDL), LDL, HDL, blood cells, and bile were collected for up to 6 days for analysis of radioactivity and mass of free and esterified cholesterol. These raw data were subjected to compartmental analysis using the SAAM program. Results in all groups corroborated net transport of free cholesterol to the liver from HDL, shown previously in fistula subjects. New findings revealed that 70% of ester was produced from free cholesterol in HDL and 30% from free cholesterol in LDL, IDL, and VLDL. No evidence was found for tissue-produced ester in plasma. There was net transfer of cholesteryl ester to VLDL and IDL from HDL and considerable exchange between LDL and HDL.Irreversible ester output was from VLDL, IDL, and LDL, but very little was from HDL, suggesting that selective and holoparticle uptakes of HDL ester are minor pathways in humans. It follows that 1) they contribute little to reverse transport, 2) very high HDL would not result from defects thereof, and 3) the clinical benefit of high HDL is likely explained by other mechanisms. Reverse transport in the subjects with bile fistula and FH was facilitated by ester output to the liver from VLDL plus IDL.

The goal of this work is to improve the inference of nonaqueous-phase contaminated source zone architectures (CSA) from field data. We follow the idea that a physically motivated model for CSA formation helps in this inference by providing relevant relationships between observables and the unknown CSA. Typical multiphase models are computationally too expensive to be applied for inverse modeling; thus, state-of-the-art CSA identification techniques do not yet use physically based CSA formation models. To overcome this shortcoming, we apply a stochastic multiphase model with reduced computational effort that can be used to generate a large ensemble of possible CSA realizations. Further, we apply a reverse transport formulation in order to accelerate the inversion of transport-related data such as downgradient aqueous-phase concentrations. We combine these approaches within an inverse Bayesian methodology for joint inversion of CSA and aquifer parameters. Because we use multiphase physics to constrain and inform the inversion, (1) only physically meaningful CSAs are inferred; (2) each conditional realization is statistically meaningful; (3) we obtain physically meaningful spatial dependencies for interpolation and extrapolation of point-like observations between the different involved unknowns and observables, and (4) dependencies far beyond simple correlation; (5) the inversion yields meaningful uncertainty bounds. We illustrate our concept by inferring three-dimensional probability distributions of DNAPL residence, contaminant mass discharge, and of other CSA characteristics. In the inference example, we use synthetic numerical data on permeability, DNAPL saturation and downgradient aqueous-phase concentration, and we substantiate our claims about the advantages of emulating a multiphase flow model with reduced computational requirement in the inversion. [ABSTRACT FROM AUTHOR]

Plasma membrane γ-aminobutyric acid (GABA) transporters (GATs) are electrogenic transport proteins that couple the cotranslocation of Na, Cl, and GABA across the plasma membrane of neurons and glia. A fundamental property of the transporter that determines its ability to concentrate GABA in cells and, hence, regulate synaptic and extra-synaptic GABA concentrations, is the ion/substrate coupling stoichiometry. Here, we scrutinized the currently accepted 2 Na:1 Cl:1 GABA stoichiometry because it is inconsistent with the measured net charge translocated per co-substrate (Na, Cl, and GABA). We expressed GAT1 and GAT3 in Xenopus laevis oocytes and utilized thermodynamic and uptake under voltage-clamp measurements to determine the stoichiometry of the GABA transporters. Voltage-clamped GAT1-expressing oocytes were internally loaded with GABA, and the reversal potential ( V) of the transporter-mediated current was recorded at different external concentrations of Na, Cl, or GABA. The shifts in V for a tenfold change in the external Na, Cl, and GABA concentration were 84 ± 4, 30 ± 1, and 29 ± 1 mV, respectively. To determine the net charge translocated per Na, Cl, and GABA, we measured substrate fluxes under voltage clamp in cells expressing GAT1 or GAT3. Charge flux to substrate flux ratios were 0.7 ± 0.1 charge/Na, 2.0 ± 0.2 charges/Cl, and 2.1 ± 0.1 charges/GABA. Altogether, our results strongly suggest a 3 Na:1 Cl:1 GABA coupling stoichiometry for the GABA transporters. The revised stoichiometry has important implications for understanding the contribution of GATs to GABAergic signaling in health and disease. [ABSTRACT FROM AUTHOR]

Serotonergic neurotransmission is terminated by reuptake of extracellular serotonin (5-HT) by the high-affinity serotonin transporter (SERT). Selective 5-HT reuptake inhibitors (SSRIs) such as fluoxetine or escitalopram inhibit SERT and are currently the principal treatment for depression and anxiety disorders. In addition, SERT is a major molecular target for psychostimulants such as cocaine and amphetamines. Amphetamine-induced transport reversal at the closely related dopamine transporter (DAT) has been shown previously to be contingent upon modulation by calmodulin kinase IIα (αCaMKII). Here, we show that not only DAT, but also SERT, is regulated by αCaMKII. Inhibition of αCaMKII activity markedly decreased amphetamine-triggered SERT-mediated substrate efflux in both cells coexpressing SERT and αCaMKII and brain tissue preparations. The interaction between SERT and αCaMKII was verified using biochemical assays and FRET analysis and colocalization of the two molecules was confirmed in primary serotonergic neurons in culture. Moreover, we found that genetic deletion of αCaMKII impaired the locomotor response of mice to 3,4-methylenedioxymethamphetamine (also known as "ecstasy") and blunted D-fenfluramine-induced prolactin release, substantiating the importance of αCaMKII modulation for amphetamine action atSERTin vivo as well. SERT-mediated substrate uptake was neither affected by inhibition of nor genetic deficiency in αCaMKII. This finding supports the concept that uptake and efflux at monoamine transporters are asymmetric processes that can be targeted separately. Ultimately, this may provide a molecular mechanism for putative drug developments to treat amphetamine addiction. [ABSTRACT FROM AUTHOR]

In monoaminergic neurons, the vesicular transporters and the plasma membrane transporters operate in a relay. Amphetamine and its congeners target this relay to elicit their actions: most amphetamines are substrates, which pervert the relay to elicit efflux of monoamines into the synaptic cleft. However, some amphetamines act as transporter inhibitors. Both compound classes elicit profound psychostimulant effects, which render them liable to recreational abuse. Currently, a surge of new psychoactive substances occurs on a global scale. Chemists bypass drug bans by ingenuous structural variations, resulting in a rich pharmacology. A credible transport model must account for their distinct mode of action and link this to subtle differences in activity and undesired, potentially deleterious effects. [ABSTRACT FROM AUTHOR]

In this paper, the viability of using micellar draw solutions (DS) for Forward Osmosis (FO) is presented for the first time. Above the critical micelle concentration (CMC) and the Krafft temperature (Tk), the monomers in a surfactant solution aggregate to form micelles, resulting in a relatively constant osmotic pressure above the CMC. These properties can be useful for a FO system in order to maintain a constant water flux and to enable draw solution regeneration at low energy. Three cationic and two anionic surfactants were studied at different concentrations as potential draw solutions for FO applications. Both flat sheet and hollow fiber FO membranes were used in the study. The study revealed that the micellar solutions generated more stable flux compared to other inorganic DS; the hypothesis of a constant flux above the CMC was valid only for concentrations slightly above the CMC, and micellar solutions behaved similar to inorganic solutes at concentrations significantly higher than the CMC. It was shown that external concentration polarization (ECP) does not have a significant effect in micellar solutions just above the CMC. Furthermore, all surfactants demonstrated between 3 and 300 times less reverse transport compared to NaCl at similar concentrations. Finally, the surfactant solute could be regenerated with recoveries as high as between 95% and 99%. It was thus verified that micellar solutions are an attractive DS for application to Forward Osmosis and potentially in the forward osmosis membrane bioreactor (FO-MBR). [ABSTRACT FROM AUTHOR]

Glutamate transporters remove the excitatory neurotransmitter glutamate from the extracellular space after neurotransmission is complete, by taking glutamate up into neurons and glia cells. As thermodynamic machines, these transporters can also run in reverse, releasing glutamate into the extracellular space. Because glutamate is excitotoxic, this transporter-mediated release is detrimental to the health of neurons and axons, and it, thus, contributes to the brain damage that typically follows a stroke. This review highlights current ideas about the molecular mechanisms underlying glutamate uptake and glutamate reverse transport. It also discusses the implications of transporter-mediated glutamate release for cellular function under physiological and patho-physiological conditions. © 2008 IUBMB IUBMB Life, 60(9): 609–619, 2008 [ABSTRACT FROM AUTHOR]

Abstract: Incubation of rat striatal slices in the absence of oxygen (anoxia), glucose (aglycemia), or oxygen plus glucose (ischemia) caused significant increases in dopamine (DA) release. Whereas anoxia decreased extracellular 3,4-dihydroxyphenylacetic acid levels by 50%, aglycemia doubled it, and ischemia returned this aglycemia-induced enhancement to its control level. Although nomifensine, a DA uptake blocker, completely protected the slices against anoxia-induced DA depletion, aglycemia-and ischemia-induced increases were not altered. Moreover, hypothermia differentially affected DA release stimulated by anoxia, aglycemia, and ischemia. Involvement of glutamate in DA release induced by each experimental condition was tested by using MK-801 and also by comparing the glutamate-induced DA release with that during anoxia, aglycemia, or ischemia. MK-801 decreased the anoxia-induced DA depletion in a dose-dependent manner. This treatment, however, showed a partial protection in aglycemic conditions but failed to improve ischemia-induced DA depletion. Like anoxia, DA release induced by exogenous glutamate was also sensitive to nomifensine and hypothermia. These results indicate that anoxia enhances DA release by a mechanism involving both the reversed DA transporter and endogenous glutamate. Partial or complete lack of effect of nomifensine, hypothermia, or MK-801 in the absence of glucose or oxygen plus glucose also suggests that experimental conditions, such as the degree of anoxia/ischemia, may alter the mechanism(s) involved in DA depletion. [ABSTRACT FROM AUTHOR]

The regulation of striatonigral and striatopallidal GABAergic neurons by glutamatergic afferents is thought to play a critical role in normal basal ganglia function. Here we report that in striatal slices about 17% of K-induced endogenous GABA release was Ca-independent and this could be blocked by a GABA transport inhibitor. Activation of N-methyl-D-aspartate (NMDA)- and quisqualate-sensitive receptors induced endogenous GABA efflux only in the presence of a GABA transaminase inhibitor; this efflux was inhibited by 60-80% with a GABA transport inhibiter. NMDA-induced GABA release was blocked by phencyclidine, Mg and CGS 19755. Quisqualate-induced GABA release was blocked completely by a combination of the metabotropic antagonist, L-AP3 and CNQX, a non-NMDA receptor antagonist. These data indicate that excitatory amino acid agonists-induced GABA release is distinct from that induced by high K depolarization. [ABSTRACT FROM AUTHOR]

Background: Much of the structure-based mechanistic understandings of the function of SLC6A neurotransmitter transporters emerged from the study of their bacterial LeuT-fold homologs. It has become evident, however, that structural differences such as the long N- and C-termini of the eukaryotic neurotransmitter transporters are involved in an expanded set of functional properties to the eukaryotic transporters. These functional properties are not shared by the bacterial homologs, which lack the structural elements that appeared later in evolution. However, mechanistic insights into some of the measured functional properties of the eukaryotic transporters that have been suggested to involve these structural elements are sparse or merely descriptive. Results: To learn how the structural elements added in evolution enable mechanisms of the eukaryotic transporters in ways not shared with their bacterial LeuT-like homologs, we focused on the human dopamine transporter (hDAT) as a prototype. We present the results of a study employing large-scale molecular dynamics simulations and comparative Markov state model analysis of experimentally determined properties of the wild-type and mutant hDAT constructs. These offer a quantitative outline of mechanisms in which a rich spectrum of interactions of the hDAT N-terminus and C-terminus contribute to the regulation of transporter function (e.g., by phosphorylation) and/or to entirely new phenotypes (e.g., reverse uptake (efflux)) that were added in evolution. Conclusions: The findings are consistent with the proposal that the size of eukaryotic neurotransmitter transporter termini increased during evolution to enable more functions (e.g., efflux) not shared with the bacterial homologs. The mechanistic explanations for the experimental findings about the modulation of function in DAT, the serotonin transporter, and other eukaryotic transporters reveal separate roles for the distal and proximal segments of the much larger N-terminus in eukaryotic transporters compared to the bacterial ones. The involvement of the proximal and distal segments — such as the role of the proximal segment in sustaining transport in phosphatidylinositol 4,5-bisphosphate-depleted membranes and of the distal segment in modulating efflux — may represent an evolutionary adaptation required for the function of eukaryotic transporters expressed in various cell types of the same organism that differ in the lipid composition and protein complement of their membrane environment. [ABSTRACT FROM AUTHOR]

The purpose of this review is to highlight recent evidence in support of a 3 Na + : 1 Cl - : 1 GABA coupling stoichiometry for plasma membrane GABA transporters (SLC6A1 , SLC6A11 , SLC6A12 , SLC6A13 ) and how the revised stoichiometry impacts our understanding of the contribution of GABA transporters to GABA homeostasis in synaptic and extrasynaptic regions in the brain under physiological and pathophysiological states. Recently, our laboratory probed the GABA transporter stoichiometry by analyzing the results of six independent measurements, which included the shifts in the thermodynamic transporter reversal potential caused by changes in the extracellular Na + , Cl - , and GABA concentrations, as well as the ratio of charge flux to substrate flux for Na + , Cl - , and GABA under voltage-clamp conditions. The shifts in the transporter reversal potential for a tenfold change in the external concentration of Na + , Cl - , and GABA were 84 ± 4, 30 ± 1, and 29 ± 1 mV, respectively. Charge flux to substrate flux ratios were 0.7 ± 0.1 charges/Na + , 2.0 ± 0.2 charges/Cl - , and 2.1 ± 0.1 charges/GABA. We then compared these experimental results with the predictions of 150 different transporter stoichiometry models, which included 1-5 Na + , 0-5 Cl - , and 1-5 GABA per transport cycle. Only the 3 Na + : 1 Cl - : 1 GABA stoichiometry model correctly predicts the results of all six experimental measurements. Using the revised 3 Na + : 1 Cl - : 1 GABA stoichiometry, we propose that the GABA transporters mediate GABA uptake under most physiological conditions. Transporter-mediated GABA release likely takes place under pathophysiological or extreme physiological conditions.