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A retinoic acid-enhanced, multicellular human blood-brain barrier model derived from stem cell sources.
Lippmann ES, Al-Ahmad A, Azarin SM, Palecek SP, Shusta EV
(2014) Sci Rep 4: 4160
MeSH Terms: Astrocytes, Blood-Brain Barrier, Brain, Cell Differentiation, Cells, Cultured, Coculture Techniques, Electric Conductivity, Endothelium, Vascular, Humans, Models, Biological, Neural Stem Cells, Neurons, Pericytes, Pluripotent Stem Cells, Tretinoin
Show Abstract · Added August 19, 2015
Blood-brain barrier (BBB) models are often used to investigate BBB function and screen brain-penetrating therapeutics, but it has been difficult to construct a human model that possesses an optimal BBB phenotype and is readily scalable. To address this challenge, we developed a human in vitro BBB model comprising brain microvascular endothelial cells (BMECs), pericytes, astrocytes and neurons derived from renewable cell sources. First, retinoic acid (RA) was used to substantially enhance BBB phenotypes in human pluripotent stem cell (hPSC)-derived BMECs, particularly through adherens junction, tight junction, and multidrug resistance protein regulation. RA-treated hPSC-derived BMECs were subsequently co-cultured with primary human brain pericytes and human astrocytes and neurons derived from human neural progenitor cells (NPCs) to yield a fully human BBB model that possessed significant tightness as measured by transendothelial electrical resistance (~5,000 Ωxcm(2)). Overall, this scalable human BBB model may enable a wide range of neuroscience studies.
0 Communities
1 Members
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15 MeSH Terms
Effects of pressure and electrical charge on macromolecular transport across bovine lens basement membrane.
Ferrell N, Cameron KO, Groszek JJ, Hofmann CL, Li L, Smith RA, Bian A, Shintani A, Zydney AL, Fissell WH
(2013) Biophys J 104: 1476-84
MeSH Terms: Animals, Basement Membrane, Biological Transport, Cattle, Electric Conductivity, Lens, Crystalline, Macromolecular Substances, Permeability, Pressure
Show Abstract · Added August 21, 2013
Molecular transport through the basement membrane is important for a number of physiological functions, and dysregulation of basement membrane architecture can have serious pathological consequences. The structure-function relationships that govern molecular transport in basement membranes are not fully understood. The basement membrane from the lens capsule of the eye is a collagen IV-rich matrix that can easily be extracted and manipulated in vitro. As such, it provides a convenient model for studying the functional relationships that govern molecular transport in basement membranes. Here we investigate the effects of increased transmembrane pressure and solute electrical charge on the transport properties of the lens basement membrane (LBM) from the bovine eye. Pressure-permeability relationships in LBM transport were governed primarily by changes in diffusive and convective contributions to solute flux and not by pressure-dependent changes in intrinsic membrane properties. The solute electrical charge had a minimal but statistically significant effect on solute transport through the LBM that was opposite of the expected electrokinetic behavior. The observed transport characteristics of the LBM are discussed in the context of established membrane transport modeling and previous work on the effects of pressure and electrical charge in other basement membrane systems.
Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.
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9 MeSH Terms
Influence of body parameters on gastric bioelectric and biomagnetic fields in a realistic volume conductor.
Kim JH, Pullan AJ, Bradshaw LA, Cheng LK
(2012) Physiol Meas 33: 545-56
MeSH Terms: Adipose Tissue, Electric Conductivity, Electrodes, Electrophysiological Phenomena, Humans, Magnetic Fields, Models, Anatomic, Stomach, Torso
Show Abstract · Added May 29, 2014
Electrogastrograms (EGG) and magnetogastrograms (MGG) provide two complementary methods for non-invasively recording electric or magnetic fields resulting from gastric electrical slow wave activity. It is known that EGG signals are relatively weak and difficult to reliably record while magnetic fields are, in theory, less attenuated by the low-conductivity fat layers present in the body. In this paper, we quantified the effects of fat thickness and conductivity values on resultant magnetic and electric fields using anatomically realistic torso models and trains of dipole sources reflecting recent experimental results. The results showed that when the fat conductivity was increased, there was minimal change in both potential and magnetic fields. However, when the fat conductivity was reduced, the magnetic fields were largely unchanged, but electric potentials had a significant change in patterns and amplitudes. When the thickness of the fat layer was increased by 30 mm, the amplitude of the magnetic fields decreased 10% more than potentials but magnetic field patterns were changed about four times less than potentials. The ability to localize the underlying sources from the magnetic fields using a surface current density measure was altered by less than 2 mm when the fat layer was increased by 30 mm. In summary, results confirm that MGG provides a favorable method over EGG when there are uncertain levels of fat thickness or conductivity.
© 2012 Institute of Physics and Engineering in Medicine
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2 Members
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9 MeSH Terms
Effect of direct current dielectrophoresis on the trajectory of a non-conducting colloidal sphere in a bent pore.
House DL, Luo H
(2011) Electrophoresis 32: 3277-85
MeSH Terms: Colloids, Computer Simulation, Electric Conductivity, Electromagnetic Fields, Electrophoresis, Microfluidics, Microspheres, Models, Chemical, Particle Size, Reproducibility of Results
Show Abstract · Added May 29, 2014
Dielectrophoresis has shown a wide range of applications in microfluidic devices. Force approximations utilizing the point-dipole method in dielectrophoresis have provided convenient predictions for particle motion by neglecting interactions between the particle and its surrounding electric and flow fields. The validity of this approach, however, is unclear when the particle size is comparable to the characteristic length of the channel and when the particle is in close proximity to the channel wall. To address this issue, we apply an accurate numerical approach based on the boundary-element method (BEM) to solve the coupled electric field, flow, and particle motion. This method can handle much closer particle-wall distances than the other numerical approaches such as the finite-element method. Using the BEM and integrating the Maxwell stress tensor, we simulate an electrokinetic, spherical particle moving within a bent cylindrical pore to investigate how the dielectrophoretic force affects the particle's trajectory. In the simulation, both the particle and the channel wall are non-conducting, and the electric double layers adjacent to the solid surfaces are assumed to be thin with respect to the particle radius and particle-wall gap. The results show that as the particle comes close to the wall, its finite size has an increasingly important effect on its own transient motion and the point-dipole approximation may lead to significant error.
Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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10 MeSH Terms
Effects of volume conductor and source configuration on simulated magnetogastrograms.
Komuro R, Qiao W, Pullan AJ, Cheng LK
(2010) Phys Med Biol 55: 6881-95
MeSH Terms: Electric Conductivity, Humans, Magnetics, Models, Biological, Stomach
Show Abstract · Added April 26, 2016
Recordings of the magnetic fields (MFs) arising from gastric electrical activity (GEA) have been shown to be able to distinguish between normal and certain abnormal GEA. Mathematical models provide a powerful tool for revealing the relationship between the underlying GEA and the resultant magnetogastrograms (MGGs). However, it remains uncertain the relative contributions that different volume conductor and dipole source models have on the resultant MFs. In this study, four volume conductor models (free space, sphere, half space and an anatomically realistic torso) and two dipole source configurations (containing 320 moving dipole sources and a single equivalent moving dipole source) were used to simulate the external MFs. The effects of different volume conductor models and dipole source configurations on the MF simulations were examined. The half space model provided the best approximation of the MFs produced by the torso model in the direction normal to the coronal plane. This was despite the fact that the half space model does not produce secondary sources, which have been shown to contribute up to 50% of the total MFs when an anatomically realistic torso model was used. We conclude that a realistic representation of the volume conductor and a detailed dipole source model are likely to be necessary when using a model-based approach for interpreting MGGs.
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5 MeSH Terms
Activation of swelling-activated chloride current by tumor necrosis factor-alpha requires ClC-3-dependent endosomal reactive oxygen production.
Matsuda JJ, Filali MS, Moreland JG, Miller FJ, Lamb FS
(2010) J Biol Chem 285: 22864-73
MeSH Terms: Animals, Cell Line, Chloride Channels, Chlorides, Electric Conductivity, Endosomes, Hydrogen Peroxide, Mice, Reactive Oxygen Species, Thrombin, Tumor Necrosis Factor-alpha
Show Abstract · Added February 22, 2016
ClC-3 is a Cl(-)/H(+) antiporter required for cytokine-induced intraendosomal reactive oxygen species (ROS) generation by Nox1. ClC-3 current is distinct from the swelling-activated chloride current (ICl(swell)), but overexpression of ClC-3 can activate currents that resemble ICl(swell). Because H(2)O(2) activates ICl(swell) directly, we hypothesized that ClC-3-dependent, endosomal ROS production activates ICl(swell). Whole-cell perforated patch clamp methods were used to record Cl(-) currents in cultured aortic vascular smooth muscle cells from wild type (WT) and ClC-3 null mice. Under isotonic conditions, tumor necrosis factor-alpha (TNF-alpha) (10 ng/ml) activated outwardly rectifying Cl(-) currents with time-dependent inactivation in WT but not ClC-3 null cells. Inhibition by tamoxifen (10 microm) and by hypertonicity (340 mosm) identified them as ICl(swell). ICl(swell) was also activated by H(2)O(2) (500 microm), and the effect of TNF-alpha was completely inhibited by polyethylene glycol-catalase. ClC-3 expression induced ICl(swell) in ClC-3 null cells in the absence of swelling or TNF-alpha, and this effect was also blocked by catalase. ICl(swell) activation by hypotonicity (240 mosm) was only partially inhibited by catalase, and the size of these currents did not differ between WT and ClC-3 null cells. Disruption of endosome trafficking with either mutant Rab5 (S34N) or Rab11 (S25N) inhibited TNF-alpha-mediated activation of ICl(swell). Thrombin also activates ROS production by Nox1 but not in endosomes. Thrombin caused H(2)O(2)-dependent activation of ICl(swell), but this effect was not ClC-3- or Rab5-dependent. Thus, activation of ICl(swell) by TNF-alpha requires ClC-3-dependent endosomal H(2)O(2) production. This demonstrates a functional link between two distinct anion currents, ClC-3 and ICl(swell).
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11 MeSH Terms
Novel selective allosteric activator of the M1 muscarinic acetylcholine receptor regulates amyloid processing and produces antipsychotic-like activity in rats.
Jones CK, Brady AE, Davis AA, Xiang Z, Bubser M, Tantawy MN, Kane AS, Bridges TM, Kennedy JP, Bradley SR, Peterson TE, Ansari MS, Baldwin RM, Kessler RM, Deutch AY, Lah JJ, Levey AI, Lindsley CW, Conn PJ
(2008) J Neurosci 28: 10422-33
MeSH Terms: Allosteric Site, Amyloid, Amyloid beta-Protein Precursor, Animals, Antipsychotic Agents, Benzimidazoles, CHO Cells, Cricetinae, Cricetulus, Dose-Response Relationship, Drug, Electric Conductivity, Hippocampus, In Vitro Techniques, Male, Patch-Clamp Techniques, Piperidines, Protein Processing, Post-Translational, Pyramidal Cells, Rats, Rats, Sprague-Dawley, Receptor, Muscarinic M1, Receptors, Dopamine D2, Receptors, N-Methyl-D-Aspartate, Synaptic Transmission, Transfection
Show Abstract · Added March 5, 2014
Recent studies suggest that subtype-selective activators of M(1)/M(4) muscarinic acetylcholine receptors (mAChRs) may offer a novel approach for the treatment of psychotic symptoms associated with schizophrenia and Alzheimer's disease. Previously developed muscarinic agonists have provided clinical data in support of this hypothesis, but failed in clinical development because of a lack of true subtype specificity and adverse effects associated with activation of other mAChR subtypes. We now report characterization of a novel highly selective agonist for the M(1) receptor with no agonist activity at any of the other mAChR subtypes, termed TBPB [1-(1'-2-methylbenzyl)-1,4'-bipiperidin-4-yl)-1H-benzo[d]imidazol-2(3H)-one]. Mutagenesis and molecular pharmacology studies revealed that TBPB activates M(1) through an allosteric site rather than the orthosteric acetylcholine binding site, which is likely critical for its unprecedented selectivity. Whole-cell patch-clamp recordings demonstrated that activation of M(1) by TBPB potentiates NMDA receptor currents in hippocampal pyramidal cells but does not alter excitatory or inhibitory synaptic transmission, responses thought to be mediated by M(2) and M(4). TBPB was efficacious in models predictive of antipsychotic-like activity in rats at doses that did not produce catalepsy or peripheral adverse effects of other mAChR agonists. Finally, TBPB had effects on the processing of the amyloid precursor protein toward the non-amyloidogenic pathway and decreased Abeta production in vitro. Together, these data suggest that selective activation of M(1) may provide a novel approach for the treatment of symptoms associated with schizophrenia and Alzheimer's disease.
1 Communities
4 Members
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25 MeSH Terms
Odor coding in the maxillary palp of the malaria vector mosquito Anopheles gambiae.
Lu T, Qiu YT, Wang G, Kwon JY, Rutzler M, Kwon HW, Pitts RJ, van Loon JJ, Takken W, Carlson JR, Zwiebel LJ
(2007) Curr Biol 17: 1533-44
MeSH Terms: Animals, Anopheles, Carbon Dioxide, Electric Conductivity, Insect Proteins, Insect Vectors, Malaria, Octanols, Odorants, Olfactory Receptor Neurons, Smell
Show Abstract · Added May 27, 2014
BACKGROUND - Many species of mosquitoes, including the major malaria vector Anopheles gambiae, utilize carbon dioxide (CO(2)) and 1-octen-3-ol as olfactory cues in host-seeking behaviors that underlie their vectorial capacity. However, the molecular and cellular basis of such olfactory responses remains largely unknown.
RESULTS - Here, we use molecular and physiological approaches coupled with systematic functional analyses to define the complete olfactory sensory map of the An. gambiae maxillary palp, an olfactory appendage that mediates the detection of these compounds. In doing so, we identify three olfactory receptor neurons (ORNs) that are organized in stereotyped triads within the maxillary-palp capitate-peg-sensillum population. One ORN is CO(2)-responsive and characterized by the coexpression of three receptors that confer CO(2) responses, whereas the other ORNs express characteristic odorant receptors (AgORs) that are responsible for their in vivo olfactory responses.
CONCLUSIONS - Our results describe a complete and highly concordant map of both the molecular and cellular olfactory components on the maxillary palp of the adult female An. gambiae mosquito. These results also facilitate the understanding of how An. gambiae mosquitoes sense olfactory cues that might be exploited to compromise their ability to transmit malaria.
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11 MeSH Terms
The quality of water from protected springs in Katwe and Kisenyi parishes, Kampala city, Uganda.
Haruna R, Ejobi F, Kabagambe EK
(2005) Afr Health Sci 5: 14-20
MeSH Terms: Cross-Sectional Studies, Electric Conductivity, Humans, Hydrogen-Ion Concentration, Nitrates, Temperature, Uganda, Waste Management, Water Microbiology, Water Pollution, Water Supply
Show Abstract · Added April 24, 2015
BACKGROUND - In the sub-urban areas of Kampala city, springs are a major source of water for domestic use. Though spring water is considered to be aesthetically acceptable for domestic use, presence of poorly designed pit latrines, poor solid waste management as well as poor and inadequate spring protection, may lead to contamination of spring water with pathogenic bacteria.
OBJECTIVES - The objectives of the study were to examine the bacteriological quality of water from ten springs in Katwe and Kisenyi parishes of Kampala, and to identify and quantify risks for spring water contamination with faecal bacteria.
METHODS - A cross-sectional sanitary risk assessment using a standardised format was carried out in ten randomly selected springs in the parishes of Katwe and Kisenyi parishes in Kampala. A total of 80 samples of water from these springs were collected from December 2001 to March 2002. The samples were analysed for indicators of faecal contamination: total coliforms, faecal coliforms and faecal streptococci. Physico-chemical parameters were measured.
RESULTS - Aggregate qualitative sanitary risk scores ranged from medium to high. The total coliform counts in 90% of the samples exceeded the WHO guideline for drinking water. All the samples had faecal coliform counts above the WHO guideline. A strong correlation (r2= 887) was observed between the median faecal coliform counts and the sanitary risk score. Sixty percent of the samples had nitrate levels above the WHO recommended limit. There was no correlation between the levels of chlorides and nitrates and levels of indicators of faecal bacterial contamination.
CONCLUSIONS - The sanitary risk assessment score is a reliable tool for predicting the likely levels of bacterial contamination of spring water. Water from the ten protected springs studied is unsuitable for drinking without treatment.
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11 MeSH Terms
Rapid stimulation causes electrical remodeling in cultured atrial myocytes.
Yang Z, Shen W, Rottman JN, Wikswo JP, Murray KT
(2005) J Mol Cell Cardiol 38: 299-308
MeSH Terms: 4-Aminopyridine, Action Potentials, Animals, Calcium, Cell Death, Cells, Cultured, Down-Regulation, Electric Conductivity, Electric Stimulation, Heart Atria, Mice, Myocytes, Cardiac, Patch-Clamp Techniques, Time Factors
Show Abstract · Added December 22, 2010
OBJECTIVE - Rapid stimulation causes electrical remodeling in the intact atrium, with shortening of action potential duration (APD), down-regulation of L-type Ca2+ currents (I(Ca,L)), and increased vulnerability to atrial fibrillation (AF). The essential elements required for this process are currently unknown. We tested the hypothesis that rapid stimulation of cardiomyocytes in vitro is sufficient to recapitulate the remodeling process, and that atrial cells subjected to rapid pacing in culture would display changes similar to those that occur in vivo.
METHODS - Atrial (HL-1) cells were cultured in the presence of rapid field stimulation (300 beats per min) for 24 h. Action potentials and ionic currents were recorded from stimulated cells, as well as control cells cultured in parallel, using whole-cell voltage-clamp techniques.
RESULTS - Rapid stimulation of atrial cells for 24 h significantly shortened APD. HL-1 cells displayed both I(Ca,L) blocked by nimodipine, and T-type Ca2+ currents (I(Ca,T)) sensitive to mibefradil. Rapid activation in culture caused down-regulation of I(Ca,L), while I(Ca,T) was similarly reduced. Multiple outward currents were present in response to a depolarizing voltage-clamp protocol, and rapid pacing resulted in up-regulation of the rapidly-activating delayed rectifier K+ current, I(Kr).
CONCLUSIONS - Rapid stimulation of atrial cells in culture produces electrical remodeling, recapitulating principal phenotypic features of atrial tachycardia remodeling in vivo. Our results demonstrate that an important component of this process is cell autonomous, given that in vivo conditions are not required for the development of electrical remodeling.
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14 MeSH Terms