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1.
J Biomech Eng ; 138(2): 021015, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26632258

ABSTRACT

Intraocular pressure (IOP) induced strains in the peripapillary sclera may play a role in glaucoma progression. Using inflation testing and ultrasound speckle tracking, the 3D strains in the peripapillary sclera were measured in nine human donor globes. Our results showed that the peripapillary sclera experienced through-thickness compression and meridional stretch during inflation, while minimal circumferential dilation was observed when IOP was increased from 10 to 19 mmHg. The maximum shear was primarily oriented in the through-thickness, meridional cross sections and had a magnitude slightly larger than the first principal strain. The tissue volume had minimal overall change, confirming near-incompressibility of the sclera. Substantial strain heterogeneity was present in the peripapillary region, with local high strain areas likely corresponding to structural heterogeneity caused by traversing blood vessels. These 3D strain characteristics provide new insights into the biomechanical responses of the peripapillary sclera during physiological increases of IOP. Future studies are needed to confirm these findings and investigate the role of these biomechanical characteristics in ocular diseases.


Subject(s)
Imaging, Three-Dimensional , Sclera/diagnostic imaging , Sclera/physiology , Stress, Mechanical , Adult , Aged , Biomechanical Phenomena , Compressive Strength , Female , Humans , Intraocular Pressure , Male , Materials Testing , Middle Aged , Optic Disk/diagnostic imaging , Optic Disk/physiology , Shear Strength , Surface Properties , Tensile Strength , Ultrasonography , Young Adult
2.
Environ Sci Pollut Res Int ; 21(17): 10194-204, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24859701

ABSTRACT

In situ metal stabilisation by amendments has been demonstrated as an appealing low-cost remediation strategy for contaminated soil. This study investigated the short-term leaching behaviour and long-term stability of As and Cu in soil amended with coal fly ash and/or green waste compost. Locally abundant inorganic (limestone and bentonite) and carbonaceous (lignite) resources were also studied for comparison. Column leaching experiments revealed that coal fly ash outperformed limestone and bentonite amendments for As stabilisation. It also maintained the As stability under continuous leaching of acidic solution, which was potentially attributed to high-affinity adsorption, co-precipitation, and pozzolanic reaction of coal fly ash. However, Cu leaching in the column experiments could not be mitigated by any of these inorganic amendments, suggesting the need for co-addition of carbonaceous materials that provides strong chelation with oxygen-containing functional groups for Cu stabilisation. Green waste compost suppressed the Cu leaching more effectively than lignite due to the difference in chemical composition and dissolved organic matter. After 9-month soil incubation, coal fly ash was able to minimise the concentrations of As and Cu in the soil solution without the addition of carbonaceous materials. Nevertheless, leachability tests suggested that the provision of green waste compost and lignite augmented the simultaneous reduction of As and Cu leachability in a fairly aggressive leaching environment. These results highlight the importance of assessing stability and remobilisation of sequestered metals under varying environmental conditions for ensuring a plausible and enduring soil stabilisation.


Subject(s)
Arsenic/chemistry , Coal Ash/chemistry , Copper/chemistry , Soil Pollutants/chemistry , Soil/chemistry , Adsorption , Arsenic/analysis , Coal , Copper/analysis , Environmental Pollution/prevention & control , Soil Pollutants/analysis
3.
J Glaucoma ; 23(8): 577-8, 2014.
Article in English | MEDLINE | ID: mdl-23632405

ABSTRACT

PURPOSE: To report a unique case of delayed topiramate-induced bilateral angle closure. METHODS: A case report of a 40-year-old woman who experienced bilateral acute angle-closure glaucoma and induced myopia on a standard and consistent dose of topiramate for 262 days before the onset of symptoms. Initial and follow-up examination findings were reviewed. RESULTS: Initial examination revealed finger counting at one foot OU, intraocular pressures of 38 mm Hg OD and 37 mm Hg OS, and bilaterally closed angles with no view of anterior trabecular meshwork (Shaffer grade 0). Discontinuation of topiramate and administration of topical drops were able to break the attack. At 3 months of follow-up, the patient was 20/25 OU with deep angles open to the scleral spur (Shaffer grade III) and had intraocular pressures of 14 mm Hg in both the eyes. CONCLUSIONS: This case provides evidence of delayed bilateral angle closure from topiramate and raises questions as to the cellular mechanism of topiramate-induced angle closure, both immediate and delayed.


Subject(s)
Fructose/analogs & derivatives , Glaucoma, Angle-Closure/chemically induced , Adult , Female , Fructose/administration & dosage , Fructose/adverse effects , Humans , Myopia/chemically induced , Topiramate
4.
J Glaucoma ; 23(2): 69-74, 2014 Feb.
Article in English | MEDLINE | ID: mdl-23603825

ABSTRACT

PURPOSE: To study differences in corneal biomechanical properties between primary open-angle glaucoma (POAG) and central corneal thickness (CCT)-matched control subjects and their effect on intraocular pressure (IOP) measurement. METHODS: Thirteen eyes of 13 POAG subjects and 15 eyes of 15 normal subjects underwent corneal topography; IOP using Goldmann applanation tonometry (GAT), dynamic contour tonometry (DCT), and corneal compensated IOP (IOPcc) using the Reichert ocular response analyzer (ORA); corneal hysteresis; and CCT. Results from POAG and control eyes were then compared using t tests. RESULTS: Ages in the POAG group were slightly greater than that in the control group. CCT was closely matched between groups. Significant differences were found between GAT versus DCT and GAT versus IOPcc within both groups: Mean GAT IOP was not significantly different between POAG and controls, whereas mean DCT IOP did show a significant difference between groups as did mean IOPcc. The delta differences, GATΔDCT and GATΔIOPcc, were of greater magnitude in POAG subjects when compared with controls. Corneal hysteresis was significantly lower in POAG subjects. CONCLUSIONS: The delta differences between GAT and newer measures of IOP are greater in magnitude in patients with POAG than in the normal controls, independent of CCT. This is likely due to differences in the corneal biomechanical properties with POAG corneas being softer than healthy corneas, which causes greater underestimation of IOP by GAT in POAG than controls. Underestimation of IOP could affect treatment decisions and outcomes in POAG.


Subject(s)
Cornea/physiology , Elasticity/physiology , Glaucoma, Open-Angle/physiopathology , Intraocular Pressure/physiology , Biomechanical Phenomena/physiology , Female , Gonioscopy , Humans , Male , Middle Aged , Prospective Studies , Tonometry, Ocular , Visual Fields/physiology
5.
Chemosphere ; 93(11): 2839-47, 2013 Nov.
Article in English | MEDLINE | ID: mdl-24144464

ABSTRACT

Utilising locally available industrial by-products for in situ metal stabilisation presents a low-cost remediation approach for contaminated soil. This study explored the potential use of inorganic (acid mine drainage (AMD) sludge and zero-valent iron) and carbonaceous materials (green waste compost, manure compost, and lignite) for minimising the environmental risks of As and Cu at a timber treatment site. After 9-month soil incubation, significant sequestration of As and Cu in soil solution was accomplished by AMD sludge, on which adsorption and co-precipitation could take place. The efficacy of AMD sludge was comparable to that of zero-valent iron. There was marginal benefit of adding carbonaceous materials. However, in a moderately aggressive environment (Toxicity Characteristic Leaching Procedure), AMD sludge only suppressed the leachability of As but not Cu. Therefore, the provision of compost and lignite augmented the simultaneous reduction of Cu leachability, probably via surface complexation with oxygen-containing functional groups. Under continuous acid leaching in column experiments, combined application of AMD sludge with compost proved more effective than AMD sludge with lignite. This was possibly attributed to the larger amount of dissolved organic matter with aromatic moieties from lignite, which may enhance Cu and As mobility. Nevertheless, care should be taken to mitigate ecological impact associated with short-term substantial Ca release and continuous release of Al at a moderate level under acid leaching. This study also articulated the engineering implications and provided recommendations for field deployment, material processing, and assessment framework to ensure an environmentally sound application of reactive materials.


Subject(s)
Environmental Restoration and Remediation/methods , Mining , Refuse Disposal/methods , Soil Pollutants/chemistry , Soil/chemistry , Water Pollutants, Chemical/chemistry , Coal , Sewage/chemistry , Soil Pollutants/analysis , Water Pollutants, Chemical/analysis
6.
Invest Ophthalmol Vis Sci ; 54(12): 7215-22, 2013 Nov 01.
Article in English | MEDLINE | ID: mdl-24130185

ABSTRACT

PURPOSE: This study tested the hypothesis that intraocular pressure (IOP) elevations, induced by controlled increase of intraocular volume, are correlated with the biomechanical responses of the posterior sclera. METHODS: Porcine globes were tested within 48 hours postmortem. The first group of globes (n = 11) was infused with 15 µL of phosphate-buffered saline at three different rates to investigate rate-dependent IOP elevations. The second group (n = 16) was first infused at the fast rate and then underwent inflation tests to investigate the relationship between IOP elevations (ΔIOP) and scleral strains. The strains in the superotemporal region of the posterior sclera were measured by ultrasound speckle tracking. Linear regression was used to examine the association between ΔIOP due to micro-volumetric infusion and the scleral strains at a specific inflation pressure. RESULTS: The average ΔIOP was 14.9 ± 4.3 mm Hg for the infusion of 15 µL in 1 second. The ΔIOP was greater for the faster infusion rates but highly correlated across different rates (P < 0.001). A significant negative association was found between the ΔIOP and the tangential strains in both the circumferential (R(2) = 0.54, P = 0.003) and meridian (R(2) = 0.53, P = 0.002) directions in the posterior sclera. CONCLUSIONS: This study showed a substantial increase in IOP, with a large intersubject variance during micro-volumetric change. A stiffer response of the sclera was associated with larger IOP spikes, providing experimental evidence linking corneoscleral biomechanics to IOP fluctuation. In vivo measurement of corneoscleral biomechanics may help better predict the dynamic profile of IOP.


Subject(s)
Intraocular Pressure/physiology , Sclera/physiology , Stress, Mechanical , Animals , Biomechanical Phenomena , Ocular Hypertension/physiopathology , Regression Analysis , Sclera/diagnostic imaging , Swine , Ultrasonography
8.
Environ Technol ; 34(21-24): 3177-82, 2013.
Article in English | MEDLINE | ID: mdl-24617077

ABSTRACT

This study aims to investigate a new and sustainable approach for the reuse of industrial by-products from wastewater treatment. The dairy industry produces huge volumes of wastewater, characterized by high levels of phosphate that can result in eutrophication and degradation of aquatic ecosystems. This study evaluated the application of acid mine drainage (AMD) sludge, coal fly ash, and lignite as low-cost adsorbents for the removal of phosphate from dairy wastewater. Material characterization using X-ray fluorescence, X-ray diffraction, and Brunauer-Emmett-Teller surface area analysis revealed significant amounts of crystalline/amorphous Fe/Al/Si/Ca-based minerals and large surface areas of AMD sludge and fly ash. Batch adsorption isotherms were best described using the Freundlich model. The Freundlich distribution coefficients were 13.7 mg(0.577) L(0.423) g(-1) and 16.9 mg(0.478) L(0.522) g(-1) for AMD sludge and fly ash, respectively, and the nonlinearity constants suggested favourable adsorption for column applications. The breakthrough curves of fixed-bed columns, containing greater than 10 wt% of the waste materials (individual or composite blends) mixed with sand, indicated that phosphate breakthrough did not occur within 100 pore volumes while the cumulative removal was 522 and 490 mg kg(-1) at 10 wt% AMD sludge and 10 wt% fly ash, respectively. By contrast, lignite exhibited negligible phosphate adsorption, possibly due to small amounts of inorganic minerals suitable for phosphate complexation and limited surface area. The results suggest that both AMD sludge and fly ash were potentially effective adsorbents if employed individually at a ratio of 10 wt% or above for column application.


Subject(s)
Acids/chemistry , Coal Ash/chemistry , Conservation of Natural Resources/methods , Phosphates/isolation & purification , Sewage/chemistry , Wastewater/analysis , Water Purification/methods , Dairying , Industrial Waste/prevention & control , Mining , Phosphates/chemistry , Water Quality
9.
Invest Ophthalmol Vis Sci ; 53(3): 1397-405, 2012 Mar 15.
Article in English | MEDLINE | ID: mdl-22297494

ABSTRACT

PURPOSE: To experimentally examine the effect of increased corneal stiffness on Goldmann applanation tonometry (GAT) and Tono-Pen (Reichert, Inc., Depew, NY) measurements of intraocular pressure (IOP) in a canine eye model. METHODS: Twenty globes were recovered from 10 dogs with no known diseases. For each dog, corneal stiffening was induced in one eye with glutaraldehyde/phosphate buffered saline (PBS) immersion while the other cornea was immersed in PBS only. Acoustic impedance was measured before and after treatment in all eyes. After treatment, IOP was measured by GAT and Tono-Pen at true pressures of 10, 15, 20, 30, and 40 mm Hg. The corneas were then dissected for uniaxial tensile testing. The GAT/Tono-Pen readings, corneal stiffness (measured by ultrasound and tensile tests), and corneal thickness were compared between the two groups. The correlations between GAT/Tono-Pen readings and corneal stiffness were evaluated. RESULTS: Acoustic impedance significantly increased after glutaraldehyde treatment (P < 0.01). Secant modulus at 1% strain was significantly higher in corneas treated with glutaradehyde/PBS than those treated with PBS only (P < 0.01). GAT and Tono-Pen readings were significantly higher at all pressure levels (P < 0.001) in the eyes with corneal stiffening. Both corneal acoustic impedance and secant modulus were significantly correlated with GAT/Tono-Pen readings at all pressure levels (P < 0.01). CONCLUSIONS: This study provided experimental evidence that corneal stiffening significantly increases GAT and Tono-Pen readings in canine eyes. Noninvasive ultrasound measurement of acoustic impedance may be used to evaluate corneal stiffness and improve the accuracy of clinical measurements of IOP.


Subject(s)
Cornea/pathology , Intraocular Pressure/physiology , Ocular Hypertension/physiopathology , Tonometry, Ocular/methods , Animals , Dogs , Humans , Ocular Hypertension/pathology , Random Allocation
10.
Invest Ophthalmol Vis Sci ; 52(11): 7866-71, 2011 Oct 04.
Article in English | MEDLINE | ID: mdl-21896862

ABSTRACT

PURPOSE: To experimentally examine the effect of corneal modulus on Goldmann applanation tonometry (GAT) and Tono-pen (Tono-pen XL, Reichert, Inc., Depew, NY) measurements of intraocular pressure (IOP) in a canine eye model. METHODS: Twenty-one canine globes were recovered from healthy animals. IOP was controlled at 10, 15, 20, 30, and 40 mm Hg and measured by GAT and Tono-pen following standard protocols. The corneas were dissected and uniaxial tensile tests were performed on corneal strips. The correlation between GAT and Tono-pen errors and corneal secant modulus was evaluated using Pearson correlation coefficients. The influence of corneal thickness and the true pressure was also examined. RESULTS: At a true IOP of 10, 15, 20, 30, and 40 mm Hg, the GAT readings were 1.1 ± 1.0, 5.1 ± 1.5, 9.5 ± 2.0, 17.3 ± 1.6, and 25.3 ± 1.8 mm Hg, respectively. The corresponding Tono-pen readings were 7.8 ± 1.7, 12.4 ± 1.7, 16.1 ± 1.9, 22.5 ± 2.1, and 28.1 ± 2.2 mm Hg, respectively. The mean secant modulus at 1% strain of the canine corneal strips was 1.54 ± 0.43 megapascal (MPa). Corneal secant modulus was significantly correlated with GAT errors when the true IOP was 30 mm Hg (R = 0.49; P < 0.05). No significant correlation was observed between tonometric errors and corneal thickness. Both GAT and Tono-pen errors increased significantly at higher pressures (P < 0.001). CONCLUSIONS: Both GAT and Tono-pen underestimated IOP in canine eyes. There was preliminary experimental evidence for a correlation between corneal modulus and GAT in the canine eyes and a higher corneal modulus was associated with higher GAT readings at a certain pressure level. The tonometric errors appeared to be pressure-dependent.


Subject(s)
Cornea/physiology , Elastic Modulus/physiology , Intraocular Pressure/physiology , Tonometry, Ocular/instrumentation , Animals , Cornea/anatomy & histology , Dogs , Models, Animal , Tensile Strength
11.
J Glaucoma ; 19(6): 382-90, 2010 Aug.
Article in English | MEDLINE | ID: mdl-20051894

ABSTRACT

PURPOSE: To evaluate a novel contact lens-embedded pressure sensor for continuous measurement of intraocular pressure (IOP). METHODS: Repeated measurements of IOP and ocular pulse amplitude (OPA) were recorded in 12 eyes of 12 subjects in sitting and supine positions using 3 configurations of the dynamic contour tonometer: slit-lamp mounted (DCT), hand-held (HH), and contact lens-embedded sensor (CL). The IOP and OPA for each condition were compared using repeated measures ANOVA and the 95% limits of agreement were calculated. RESULTS: The sitting IOP (mean and 95% CI) for each configuration was DCT: 16.3 mm Hg (15.6 to 17.1 mm Hg), HH: 16.6 mm Hg (15.6 to 17.6 mm Hg), and CL: 15.7 mm Hg (15 to 16.3 mm Hg). The sitting OPA for each configuration was DCT: 2.4 mm Hg (2.1 to 2.6 mm Hg), HH: 2.4 mm Hg (2.1 to 2.7 mm Hg), and CL: 2.1 mm Hg (1.8 to 2.3 mm Hg). Supine IOP and OPA measurements with the CL and HH sensors were both greater than their corresponding sitting measurements, but were significantly less with the CL sensor than the HH sensor. The mean difference and 95% Limits of Agreement were smallest for the DCT and CL sensor comparisons (0.7+/-3.9 mm Hg) and widest for the CL and HH sensors (-1.9+/-7.25 mm Hg); these wider limits were attributed to greater HH measurement variability. CONCLUSIONS: The CL sensor was comparable to HH and DCT sensors with sitting subjects and is a viable method for measuring IOP and OPA. Supine measurements of IOP and OPA were greater than sitting conditions and were comparatively lower with the CL sensor. HH measurements were more variable than CL measurements and this influenced the Limits of Agreement for both sitting and supine conditions.


Subject(s)
Contact Lenses , Intraocular Pressure/physiology , Tonometry, Ocular/instrumentation , Adult , Blood Pressure/physiology , Female , Humans , Male , Posture , Reproducibility of Results , Transducers, Pressure , Young Adult
12.
Water Res ; 43(4): 961-70, 2009 Mar.
Article in English | MEDLINE | ID: mdl-19070349

ABSTRACT

Bioreactors represent an emerging technology for removing metals and sulfate commonly found in acid mine drainage (AMD). Six continuously fed anaerobic bioreactors employing organic and alkaline waste materials were operated to investigate relationships between metal and sulfate removal from AMD. Median AMD influent chemistry was 65.8mg/L Fe (49.7-113mg/L), 46.5mg/L Al (33.5-72.4mg/L) and 608mg/L sulfate (493-1007mg/L). Bioreactors containing mussel shells as an alkaline substrate amendment were more effective at removing metals and sulfate than those containing limestone. Experimental results indicated bioreactor design and operation should be dependent on treatment goals. These include 0.3mol sulfate loading/m(3)/day for sulfate removal (mean of 94.1% (87.6-98.0%), 0.4mol metals/m(3)/day for metal (mean of 99.0% (98.5-99.9%)) and partial sulfate (mean of 46.0% (39.6-57.8%)) removal and 0.8mol metals/m(3)/day for metal (mean of 98.4% (98.2-98.6%) and minimal sulfate (mean of 16.6% (11.9-19.2%)) removal. Aluminum removal efficiency was on average 1.72% (0.04-3.42%) greater than Fe during stable operating conditions.


Subject(s)
Aluminum/analysis , Bioreactors , Drainage/methods , Iron/analysis , Metals/isolation & purification , Mining , Sulfates/isolation & purification , Hydrogen-Ion Concentration , Industrial Waste , Waste Disposal, Fluid/methods , Water Pollutants, Chemical/isolation & purification , Water Purification/methods
13.
Invest Ophthalmol Vis Sci ; 49(9): 3919-26, 2008 Sep.
Article in English | MEDLINE | ID: mdl-18539936

ABSTRACT

PURPOSE: To develop a method for evaluating viscosity and elasticity of the cornea and to examine the effect that both properties have on hysteresis. METHODS: A three-component spring and dashpot model was created in Simulink in Matlab to represent the purely elastic and viscoelastic behavior of the cornea during a measurement using device called an ocular response analyzer (ORA). Values for elasticity and viscosity were varied while sinusoidal stress was applied to the model. The simulated stresses were used to determine how hysteresis is affected by the individual components of elasticity, viscosity, and maximum stress. To validate the model, high-speed photography was used to measure induced strain in a corneal phantom during ORA measurement. This measured strain was compared with the strains simulated by the model. RESULTS: When the spring in the viscoelastic portion of the model was stiffened, hysteresis decreased. When the spring in the purely elastic element was stiffened, hysteresis increased. If both springs were stiffened together, hysteresis peaked strongly as a function of the viscosity of the viscoelastic element. Below the peak value, lower elasticity was associated with higher hysteresis. Above the peak value, higher elasticity was associated with higher hysteresis. In addition, hysteresis increased as the air maximum pressure was increased. Measurements from phantom corresponded to predictions from the model. CONCLUSIONS: A viscoelastic model is presented to illustrate how changing viscosity and elasticity may affect hysteresis. Low hysteresis can be associated with either high elasticity or low elasticity, depending on the viscosity, a finding consistent with clinical reports.


Subject(s)
Cornea/physiology , Biomechanical Phenomena , Corneal Topography , Elasticity , Freezing , Humans , Intraocular Pressure , Models, Biological , Stress, Mechanical , Tonometry, Ocular , Viscosity
14.
Curr Eye Res ; 32(7-8): 625-38, 2007.
Article in English | MEDLINE | ID: mdl-17852186

ABSTRACT

OBJECTIVE: To determine the effect of low-fluence diode laser irradiation upon the fluid perfusion characteristics of cultured human trabecular meshwork cell monolayers when placed in a specially designed testing apparatus and subjected to fluid flow driven by a hydrostatic pressure gradient. METHODS: Two experimental series were conducted. In the first series, six low-fluence diode laser irradiation experiments were conducted using cultured human trabecular meshwork cell monolayers grown on filter supports. Upon reaching a steady state perfusion condition at approximately 5.0 mmHg, monolayers were irradiated at fluencies ranging from 0.2619 to 0.8571 J/cm2 using a diode laser (lambda=810 nm). Perfusion and data collection continued for 45 minutes post-irradiation, after which the monolayers were tested to determine post-experimental viability. Hydraulic conductivity values were analyzed for post-irradiation response in 2.5-minute intervals, grouped by viability. In the second series, a total of six irradiated experiments and six simultaneous nonirradiated control experiments were conducted. Fluence values of 0.3571 J/cm2 (n=3) and 0.4286 J/cm2 (n=3) were used. Hydraulic conductivity values were analyzed for post-irradiation response in 2.5-minute intervals, grouped by irradiated vs. nonirradiated control groups. RESULTS: In the first series, analysis showed that the viable monolayers exhibited a statistically significant increase in hydraulic conductivity (p<0.001) from 10 minutes post-irradiation onward. The non-viable monolayers exhibited a statistically significant decrease in hydraulic conductivity. In the second series, irradiated groups showed a significant difference (p<0.001) from nonirradiated controls from 10 minutes post-irradiation onward. CONCLUSION: Low-fluence diode laser irradiation increases hydraulic conductivity in viable perfused TM cell monolayers when compared to baseline values or simultaneous nonirradiated controls while decreasing hydraulic conductivity in nonviable monolayers.


Subject(s)
Aqueous Humor/metabolism , Endothelial Cells/radiation effects , Low-Level Light Therapy , Trabecular Meshwork/radiation effects , Biological Transport , Caco-2 Cells , Cells, Cultured , Cytoskeletal Proteins/metabolism , Dexamethasone/pharmacology , Endothelial Cells/drug effects , Endothelial Cells/metabolism , Eye Proteins/metabolism , Glucocorticoids/pharmacology , Glycoproteins/metabolism , Humans , Hydrostatic Pressure , Membrane Proteins/metabolism , Phosphoproteins/metabolism , Trabecular Meshwork/drug effects , Trabecular Meshwork/metabolism , Zonula Occludens-1 Protein
15.
Biophys J ; 87(2): 958-73, 2004 Aug.
Article in English | MEDLINE | ID: mdl-15298902

ABSTRACT

Gap junctions have traditionally been characterized as nonspecific pores between cells passing molecules up to 1 kDa in molecular mass. Nonetheless, it has become increasingly evident that different members of the connexin (Cx) family mediate quite distinct physiological processes and are often not interchangeable. Consistent with this observation, differences in permeability to natural metabolites have been reported for different connexins, although the physical basis for selectivity has not been established. Comparative studies of different members of the connexin family have provided evidence for ionic charge selectivity, but surprisingly little is known about how connexin composition affects the size of the pore. We have employed a series of Alexa dyes, which share similar structural characteristics but range in size from molecular weight 350 to 760, to probe the permeabilities and size limits of different connexin channels expressed in Xenopus oocytes. Correlated dye transfer and electrical measurements on each cell pair, in conjunction with a three-dimensional mathematical model of dye diffusion in the oocyte system, allowed us to obtain single channel permeabilities for all three dyes in six homotypic and four heterotypic channels. Cx43 and Cx32 channels passed all three dyes with similar efficiency, whereas Cx26, Cx40, and Cx45 channels showed a significant drop-off in permeability with the largest dye. Cx37 channels only showed significant permeability for the smaller two dyes, but at two- to sixfold lower levels than other connexins tested. In the heterotypic cases studied (Cx26/Cx32 and Cx43/Cx37), permeability characteristics were found to resemble the more restrictive parental homotypic channel. The most surprising finding of the study was that the absolute permeabilities calculated for all gap junctional channels in this study are, with one exception, at least 2 orders of magnitude greater than predicted purely on the basis of hindered pore diffusion. Consequently, affinity between the probes and the pore creating an energetically favorable in-pore environment, which would elevate permeant concentration within the pore and hence the flux, is strongly implicated.


Subject(s)
Cell Membrane Permeability/physiology , Connexins/metabolism , Fluorescent Dyes/chemistry , Gap Junctions/metabolism , Ion Channels/physiology , Oocytes/physiology , Spectrometry, Fluorescence/methods , Animals , Cells, Cultured , Computer Simulation , Connexins/genetics , Fluorescent Dyes/metabolism , Ion Channel Gating/physiology , Models, Biological , Models, Chemical , Particle Size , Porosity , Recombinant Proteins/metabolism , Xenopus laevis
16.
Biophys J ; 86(4): 2058-77, 2004 Apr.
Article in English | MEDLINE | ID: mdl-15041648

ABSTRACT

As ubiquitous conduits for intercellular transport and communication, gap junctional pores have been the subject of numerous investigations aimed at elucidating the molecular mechanisms underlying permeability and selectivity. Dye transfer studies provide a broadly useful means of detecting coupling and assessing these properties. However, given evidence for selective permeability of gap junctions and some anomalous correlations between junctional electrical conductance and dye permeability by passive diffusion, the need exists to give such studies a more quantitative basis. This article develops a detailed diffusion model describing experiments (reported separately) involving transport of fluorescent dye from a "donor" region to an "acceptor" region within a pair of Xenopus oocytes coupled by gap junctions. Analysis of transport within a single oocyte is used to determine the diffusion and binding characteristics of the cellular cytoplasm. Subsequent double-cell calculations then yield the intercellular junction permeability, which is translated into a single-channel permeability using concomitant measurements of intercellular conductance, and known single-channel conductances of gap junctions made up of specific connexins, to count channels. The preceding strategy, combined with use of a graded size series of Alexa dyes, permits a determination of absolute values of gap junctional permeability as a function of dye size and connexin type. Interpretation of the results in terms of pore theory suggests significant levels of dye-pore affinity consistent with the expected order of magnitude of typical (e.g., van der Waals) intermolecular attractions.


Subject(s)
Cell Communication/physiology , Cell Membrane Permeability/physiology , Gap Junctions/physiology , Models, Theoretical , Oocytes/physiology , Animals , Connexins/metabolism , Fluorescent Dyes/chemistry , Xenopus
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