Australian per caput dose from diagnostic imaging and nuclear medicine.

The largest man-made contributor to the ionising radiation dose to the Australian population is from diagnostic imaging and nuclear medicine. The last estimation of this dose was made in 2004 (1.3 mSv), this paper describes a recent re-evaluation of this dose to reflect the changes in imaging trends and technology. The estimation was calculated by summing the dose from five modalities, computed tomography (CT), general radiography/fluoroscopy, interventional procedures, mammography and nuclear medicine. Estimates were made using Australian frequency data and dose data from a range of Australian and international sources of average effective dose values. The ionising radiation dose to the Australian population in 2010 from diagnostic imaging and nuclear medicine is estimated to be 1.7 mSv (1.11 mSv CT, 0.30 mSv general radiography/fluoroscopy, 0.17 mSv interventional procedures, 0.03 mSv mammography and 0.10 mSv nuclear medicine). This exceeds the estimate of 1.5 mSv per person from natural background and cosmic radiation.

Soil [N] modulates soil C cycling in CO2-fumigated tree stands: a meta-analysis.

Under elevated atmospheric CO(2) concentrations, soil carbon (C) inputs are typically enhanced, suggesting larger soil C sequestration potential. However, soil C losses also increase and progressive nitrogen (N) limitation to plant growth may reduce the CO(2) effect on soil C inputs with time. We compiled a data set from 131 manipulation experiments, and used meta-analysis to test the hypotheses that: (1) elevated atmospheric CO(2) stimulates soil C inputs more than C losses, resulting in increasing soil C stocks; and (2) that these responses are modulated by N. Our results confirm that elevated CO(2) induces a C allocation shift towards below-ground biomass compartments. However, the increased soil C inputs were offset by increased heterotrophic respiration (Rh), such that soil C content was not affected by elevated CO(2). Soil N concentration strongly interacted with CO(2) fumigation: the effect of elevated CO(2) on fine root biomass and -production and on microbial activity increased with increasing soil N concentration, while the effect on soil C content decreased with increasing soil N concentration. These results suggest that both plant growth and microbial activity responses to elevated CO(2) are modulated by N availability, and that it is essential to account for soil N concentration in C cycling analyses.

Elevated CO(2) and elevated temperature have no effect on Douglas-fir fine-root dynamics in nitrogen-poor soil.

Here, we investigate fine-root production, mortality and standing crop of Douglas-fir (Pseudotsuga menziesii) seedlings exposed to elevated atmospheric CO(2) and elevated air temperature. We hypothesized that these treatments would increase fine-root production, but that mortality would be greater under elevated temperature, leading to a smaller increase in standing crop. Seedlings were grown in outdoor, sun-lit controlled-environment chambers containing native soil. They were exposed in a factorial design to two levels of atmospheric CO(2) and two levels of air temperature. Minirhizotron methods were used to measure fine-root length production, mortality and standing crop every 4 wk for 36 months. Neither elevated atmospheric CO(2) nor elevated air temperature affected fine-root production, mortality, or standing crop. Fine roots appeared to root deeper in the soil profile under elevated CO(2) and elevated temperature. Low soil nitrogen (N) levels apparently limited root responses to the treatments. This suggests that forests on nutrient-poor soils may exhibit limited fine-root responses to elevated atmospheric CO(2) and elevated air temperature.

Does soil CO2 efflux acclimatize to elevated temperature and CO2 during long-term treatment of Douglas-fir seedlings?

We investigated the effects of elevated soil temperature and atmospheric CO2 on soil CO2 efflux (SCE) during the third and fourth years of study. We hypothesized that elevated temperature would stimulate SCE, and elevated CO2 would also stimulate SCE with the stimulation being greater at higher temperatures. The study was conducted in sun-lit controlled-environment chambers using Douglas-fir (Pseudotsuga menziesii) seedlings grown in reconstructed litter-soil systems. We used a randomized design with two soil temperature and two atmospheric CO2 treatments. The SCE was measured every 4 wk for 18 months. Neither elevated temperature nor CO2 stimulated SCE. Elevated CO2 increased the temperature sensitivity of SCE. During the winter, the relationship between SCE and soil moisture was negative but it was positive during the summer. The seasonal patterns in SCE were associated with seasonal changes in photosynthesis and above-ground plant growth. SCE acclimatized in the high-temperature treatment, probably because of a loss of labile soil carbon. Elevated CO2 treatment increased the temperature sensitivity of SCE, probably through an increase in substrate availability.

Foliar nitrogen concentrations and natural abundance of (15)N suggest nitrogen allocation patterns of Douglas-fir and mycorrhizal fungi during development in elevated carbon dioxide concentration and temperature.

Pseudotsuga menziesii (Mirb.) Franco (Douglas-fir) seedlings were grown in a 2 x 2 factorial design in enclosed mesocosms at ambient temperature or 3.5 degrees C above ambient, and at ambient CO2 concentration ([CO2]) or 179 ppm above ambient. Two additional mesocosms were maintained as open controls. We measured the extent of mycorrhizal infection, foliar nitrogen (N) concentrations on both a weight basis (%N) and area basis (Narea), and foliar delta15N signatures (15N/14N ratios) from summer 1993 through summer 1997. Mycorrhizal fungi had colonized nearly all root tips across all treatments by spring 1994. Elevated [CO2] lowered foliar %N but did not affect N(area), whereas elevated temperature increased both foliar %N and Narea. Foliar delta15N was initially -1 per thousand and dropped by the final harvest to between -4 and -5 per thousand in the enclosed mesocosms, probably because of transfer of isotopically depleted N from mycorrhizal fungi. Based on the similarity in foliar delta15N among treatments, we conclude that mycorrhizal fungi had similar N allocation patterns across CO2 and temperature treatments. We combined isotopic and Narea data for 1993-94 to calculate fluxes of N for second- and third-year needles. Yearly N influxes were higher in second-year needles than in third-year needles (about 160 and 50% of initial leaf N, respectively), indicating greater sink strength in the younger needles. Influxes of N in second-year needles increased in response to elevated temperature, suggesting increased N supply from soil relative to plant N demands. In the elevated temperature treatments, N effluxes from third-year needles were higher in seedlings in elevated [CO2] than in ambient [CO2], probably because of increased N allocation below ground. We conclude that N allocation patterns shifted in response to the elevated temperature and [CO2] treatments in the seedlings but not in their fungal symbionts.

Advancing fine root research with minirhizotrons.

Minirhizotrons provide a nondestructive, in situ method for directly viewing and studying fine roots. Although many insights into fine roots have been gained using minirhizotrons, a review of the literature indicates a wide variation in how minirhizotrons and minirhizotron data are used. Tube installation is critical, and steps must be taken to insure good soil/tube contact without compacting the soil. Ideally, soil adjacent to minirhizotrons will mimic bulk soil. Tube installation causes some degree of soil disturbance and has the potential to create artifacts in subsequent root data and analysis. We therefore recommend a waiting period between tube installation and image collection of 6-12 months to allow roots to recolonize the space around the tubes and to permit nutrients to return to pre-disturbance levels. To make repeated observations of individual roots for the purposes of quantifying their dynamic properties (e.g. root production, turnover or lifespan), tubes should be secured to prevent movement. The frequency of image collection depends upon the root parameters being measured or calculated and the time and resources available for collecting images and extracting data. However, long sampling intervals of 8 weeks or more can result in large underestimates of root dynamic properties because more fine roots will be born and die unobserved between sampling events. A sampling interval of 2 weeks or less reduces these underestimates to acceptable levels. While short sample intervals are desirable, they can lead to a potential trade-off between the number of minirhizotron tubes used and the number of frames analyzed per tube. Analyzing fewer frames per minirhizotron tube is one way to reduce costs with only minor effects on data variation. The quality of minirhizotron data should be assessed and reported; procedures for quantifying the quality of minirhizotron data are presented here. Root length is a more sensitive metric for dynamic root properties than the root number. To make minirhizotron data from separate experiments more easily comparable, idiosyncratic units should be avoided. Volumetric units compatible with aboveground plant measures make minirhizotron-based estimates of root standing crop, production and turnover more useful. Methods for calculating the volumetric root data are discussed and an example presented. Procedures for estimating fine root lifespan are discussed.

Interactive effects of CO2 and O3 on a ponderosa pine plant/litter/soil mesocosm.

To study individual and combined impacts of two important atmospheric trace gases, CO2 and O3, on C and N cycling in forest ecosystems; a multi-year experiment using a small-scale ponderosa pine (Pinus ponderosa Laws.) seedling/soil/litter system was initiated in April 1998. The experiment was conducted in outdoor, sun-lit chambers where aboveground and belowground ecological processes could be studied in detail. This paper describes the approach and methodology used, and presents preliminary data for the first two growing seasons. CO2 treatments were ambient and elevated (ambient + 280 ppm). O3 treatments were elevated (hourly averages to 159 ppb, cumulative exposure > 60 ppb O3, SUM 06 approximately 10.37 ppm h), and a low control level (nearly all hourly averages <40 ppb. SUM 06 approximately 0.07 ppm h). Significant (P < 0.05) individual and interactive effects occurred with elevated CO2 and elevated O3. Elevated CO2 increased needle-level net photosynthetic rates over both seasons. Following the first season, the highest photosynthetic rates were for trees which had previously received elevated O3 in addition to elevated CO2. Elevated CO2 increased seedling stem diameters, with the greatest increase at low O3. Elevated CO2 decreased current year needle % N in the summer. For 1-year-old needles measured in the fall there was a decrease in % N with elevated CO2 at low O3, but an increase in % N with elevated CO2 at elevated O3. Nitrogen fixation (measured by acetylene reduction) was low in ponderosa pine litter and there were no significant CO2 or O3 effects. Neither elevated CO2 nor elevated O3 affected standing root biomass or root length density. Elevated O3 decreased the % N in coarse-fine (1-2 mm diameter) but not in fine (< 1 mm diameter) roots. Both elevated CO2 and elevated O3 tended to increase the number of fungal colony forming units (CFUs) in the AC soil horizon, and elevated O3 tended to decrease bacterial CFUs in the C soil horizon. Thus, after two growing seasons we showed interactive effects of O3 and CO2 in combination, in addition to responses to CO2 or O3 alone for a ponderosa pine plant/litter/soil system.

S-nitrosoglutathione photolysis as a novel therapy for antifibrosis in filtration surgery.

PURPOSE: To determine whether a novel peroxynitrite-based photosensitizer S-nitrosoglutathione (GSNO) can produce specific in vitro light-induced cell death of both standard animal lung and human Tenon's capsule (TC) fibroblasts and to compare this effect with that produced by the established photodynamic porphyrin precursor 5-aminolevulinic acid (ALA). METHODS: V79-4 Chinese hamster lung and human TC fibroblasts were established in tissue culture. GSNO, together with its radioactive tritiated and fluorescent dansylated derivatives, were synthesized. The labeled molecules were prepared to determine the time course of uptake into the fibroblasts. Uptake was monitored by scintillation counting for the tritiated GSNO and confocal fluorescence microscopy for the dansylated GSNO. The uptake of ALA and biosynthesis of its photosensitive product were determined by fluorescence emission spectroscopy of a separate set of fibroblasts. Once uptake was established, both cell lines were incubated with varying concentrations of GSNO or ALA as a function of time (0, 4, or 24 hours) before light exposure (200 msec pulsed visible light, 0.068 W per pulse, for 10 minutes at a distance of 10 cm). After 10 minutes of irradiation, the cells were washed and exposed to fresh tissue culture medium. The effect of the treatment was determined 24 hours later by measuring cell viability. RESULTS: A 2-minute drug treatment time (0 hours incubation) with GSNO, followed by 10 minutes of irradiation, resulted in approximately 78% of fibroblast cell death at the lowest concentration of GSNO used compared with the control, which was exposed to light, but no GSNO. The higher concentrations of GSNO, or longer drug treatment times before irradiation, did not statistically increase cell death. Maximal cell death was thus obtained using the lowest GSNO concentration (50 mM) and drug treatment time (2 minutes). In contrast, the well-established photosensitizer ALA killed only approximately 4% of cells at the lowest concentration and drug treatment time tested. At drug treatment times of 4 hours and less, increased concentrations of ALA did not produce cell death of more statistical significance. It was not until 24 hours of drug treatment that comparable amounts of cell death were produced by ALA and GSNO. In all experiments similar results were obtained with the animal lung and human TC fibroblasts, suggesting that the source of the fibroblast had no effect on the outcome. The differences in treatment effects between GSNO and ALA were statistically significant under all conditions tested. CONCLUSIONS: GSNO is able to cause light-specific cell death of human TC fibroblasts at drug treatment times (2 minutes) and irradiation times (10 minutes) that would be compatible with its use in glaucoma filtering surgery. This in vitro performance was superior to that of the well-established photosensitizer ALA, which required treatment times longer than 4 hours to approach the light-specific cell death produced by only 2 minutes of GSNO treatment.

Meta-analysis of the effect of latanoprost and brimonidine on intraocular pressure in the treatment of glaucoma.

OBJECTIVE: The purpose of this study was to indirectly quantify and compare the intraocular pressure (IOP)-lowering effects of latanoprost and brimonidine eye drops at baseline and after 3 and 6 months in the treatment of primary open-angle glaucoma. METHODS: This meta-analysis combined data from all randomized controlled trials comparing the effects on IOP of latanoprost and brimonidine treatment in adults with a baseline IOP > or =20 mm Hg. MEDLINE and EMBASE were searched for reports of the ophthalmic administration of either drug versus the other, placebo, or active therapy. Included studies reported IOP as either means or differences (with SD or SE) and sample sizes. A random-effects model was used to pool data within each drug group. As a proxy for success rates, area under the curve (AUC) was calculated for the proportion of patients having an IOP <20 mm Hg. RESULTS: One hundred fifty-five articles reporting on 158 trials were identified; 147 papers were rejected (141 were not randomized controlled trials, 5 were duplicates, and 1 had nonextractable data), leaving 9 trials from 8 articles. A total of 2152 patients were included in the meta-analysis: 597 received latanoprost, 571 received brimonidine, and the remainder received timolol or betaxolol. Baseline IOPs were similar in patients randomized to latanoprost or brimonidine (25.3 and 24.6 mm Hg, respectively). At 3 months, latanoprost and brimonidine reduced IOP by 8.4 and 6.5 mm Hg, respectively (P = 0.004 latanoprost vs brimonidine), and at 6 months by 8.0 and 6.2 mm Hg, respectively (P = 0.045). AUC was 0.834 and 0.675 at 3 months for latanoprost and brimonidine, respectively, and 0.817 and 0.715 at 6 months, respectively (both, P < 0.001). CONCLUSIONS: This indirect comparison of data from the available randomized clinical trials showed latanoprost to be statistically superior to brimonidine in reducing IOP in adults with primary open-angle glaucoma. Additional long-term, head-to-head comparisons of the efficacy, safety, and cost of latanoprost and brimonidine are needed to support and supplement these findings.

Vertical gradients in photosynthetic light response within an old-growth Douglas-fir and western hemlock canopy.

We examined needle-level light response of photosynthesis across a vertical light gradient within 45-55-m-tall western hemlock (Tsuga heterophylla (Raf.) Sarg.) and Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) trees growing in a 400-500-year-old mixed species stand. We determined: (1) whether light-saturated photosynthetic rates, light compensation points, and respiration rates varied from the upper to the lower canopy, and (2) if light-saturated photosynthetic rates, light compensation points, and respiration rates varied between Douglas-fir and western hemlock. Over a 25-m gradient from the canopy top to the lower canopy, mean light-saturated photosynthetic rates, light compensation points, and respiration rates declined in overstory Douglas-fir and western hemlock needles, paralleling a 65% decline in the mean daily photosynthetic photon flux density (PPFD). At the canopy top, increasing light-saturated photosynthetic rates relative to lower canopy needles increased carbon uptake at high PPFD. In the lower canopy, reduced respiration rates relative to upper canopy needles increased carbon uptake at low PPFD by reducing the light compensation point. At all canopy positions, western hemlock had lower mean light-saturated photosynthetic rates, light compensation points and respiration rates than Douglas-fir. As a result, western hemlock had higher net photosynthetic rates at low PPFD, but lower net photosynthetic rates at high PPFD compared with Douglas-fir.

Central corneal thickness in low-tension glaucoma.

BACKGROUND: It is possible that the intraocular pressure (IOP) is underestimated in eyes whose central cornea is thinner than normal. The objective of this study was to determine and establish the significance of central corneal thickness in patients with low-tension (normal-tension) glaucoma compared with those with chronic open-angle glaucoma (COAG) or ocular hypertension and healthy eyes. METHODS: The study was carried out from February 1998 to May 1999. Central corneal thickness was measured by ultrasonic pachymetry and IOP was measured by Goldmann applanation tonometry in 25 patients with low-tension glaucoma (untreated IOP less than 21 mm Hg with evidence of optic nerve head damage and corresponding visual field loss on automated perimetry), 80 patients with COAG (untreated IOP 21 mm Hg or greater with evidence of optic nerve head damage and corresponding visual field loss on automated perimetry), 16 patients with ocular hypertension (untreated IOP 21 mm Hg or greater, with normal optic nerve head and no history of glaucoma or elevated IOP, and normal visual field on automated perimetry) and 50 control subjects (untreated IOP less than 21 mm Hg with normal optic nerve head and no history of glaucoma or elevated IOP). Analysis with Pearson's product-moment correlation was performed to determine the correlation of IOP and central corneal thickness, and one-way analysis of variance was used to compare corneal thickness between groups. RESULTS: The central cornea was significantly thinner in the low-tension glaucoma group (mean 513.2 mu [standard deviation (SD) 26.1 mu]) than in the COAG group (mean 548.2 mu [SD 35.0 mu]) and the control group (mean 556.7 mu [SD 35.9 mu]) (p < 0.001). No significant difference in corneal thickness was found between the COAG and control groups. The ocular hypertension group had significantly thicker corneas (mean 597.5 mu [SD 23.6 mu]) than the three other groups (p < 0.001). INTERPRETATION: Patients with low-tension glaucoma may have thinner corneas than patients with COAG and healthy subjects. This results in underestimation of their IOP. Corneal thickness should be taken into account when managing these patients to avoid undertreatment.

Seasonal patterns of photosynthetic light response in Douglas-fir seedlings subjected to elevated atmospheric CO(2) and temperature.

Increases in atmospheric CO(2) concentration and temperature are predicted to increase the light response of photosynthesis by increasing light-saturated photosynthetic rates and apparent quantum yields. We examined the interactive effects of elevated atmospheric CO(2) concentration and temperature on the light response of photosynthesis in Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings. Seedlings were grown in sunlit chambers controlled to track either ambient (~400 ppm) CO(2) or ambient + 200 ppm CO(2), at ambient temperature or ambient + 4 degrees C. Photosynthetic light response curves were measured over an 18-month period beginning 32 months after treatments were initiated. Light-response curves were measured at the growth CO(2) concentration, and were used to calculate the light-saturated rate of photosynthesis, light compensation point, quantum yield and respiration rate. Elevated CO(2) increased apparent quantum yields during two of five measurement periods, but did not significantly affect light-saturated net photosynthetic rates, light compensation points or respiration rates. Elevated temperature increased all parameters. There were no significant interactions between CO(2) concentration and temperature. We conclude that down-regulation of photosynthesis occurred in the elevated CO(2) treatments such that carbon uptake at a given irradiance was similar across CO(2) treatments. In contrast, increasing temperature may substantially increase carbon uptake rates in Douglas-fir, assuming other environmental factors do not limit photosynthesis; however, it is not clear whether the increased carbon uptake will increase growth rates or be offset by increased carbon efflux through respiration.

Correlation of intraocular pressure and central corneal thickness in normal myopic eyes and after laser in situ keratomileusis.

PURPOSE: To determine the relationship of intraocular pressure (IOP) and central corneal thickness (CCT) in normal myopic eyes and after laser in situ keratomileusis (LASIK). SETTING: TLC The Windsor Laser Center, Windsor, Ontario, Canada. METHODS: Intraocular pressure measured by Goldmann applanation tonometry and CCT by ultrasonic pachymetry were determined in a group of untreated corneas of 120 patients (203 eyes) and in 50 patients (85 eyes) pre- and post-LASIK. Statistical analyses were performed with the Pearson correlation coefficient and paired Student t test. RESULTS: In the untreated group of 288 eyes, mean CCT was 544.0 microns +/- 37.3 (SD) (range 461 to 664 microns) and mean IOP, 15.6 +/- 2.7 mm Hg (range 10 to 24 mm Hg). The correlation between IOP and CCT in this group was highly significant (r = 0.44; P < .0001). The slope was 0.032 mm Hg/micron of CCT or an approximate decrease of 1 mm Hg, for a reduction in CCT of 31.3 microns. In the post-LASIK group, mean CCT dropped approximately 73.0 microns to 479.5 +/- 41.2 microns (range 408 to 503 microns) and IOP dropped to a mean of 13.6 +/- 3.3 mm Hg (range 7 to 22 mm Hg). A significant correlation was found between IOP and CCT after LASIK (r = 0.33; P < .002). The difference between the mean pre- and post-LASIK measurements of applanation IOP was 2.5 mm Hg, which was significant (P < .0001). The post-LASIK slope was 0.027 mm Hg/micron, or a decrease of 1.0 mm Hg per 37.8 microns reduction in CCT. CONCLUSION: Central corneal thickness is an important variable in the evaluation of applanation IOP and should be included in the assessment of any case of potential glaucoma or ocular hypertension, particularly in eyes with previous photoablative refractive surgery.

Stability of the antiproliferative effect of mitomycin-C after reconstitution.

PURPOSE: The purpose of this study was to determine the long-term stability of the antiproliferative effect of mitomycin-C (MMC) following reconstitution. METHODS: Identical MMC preparations were reconstituted from the crystalline form and then stored at room temperature or 4 degrees C. Cultured human Tenon's fibroblasts were exposed to MMC (0.5 mg/ml) for 2.5 min at 0, 3, 7, 10, 14, 21, 28, 35, and 42 days following reconstitution. After removal of the drug, fibroblast proliferation was measured by tritiated thymidine uptake. RESULTS: The percentage of inhibition was maintained at > 85% for both the room temperature and the 4 degrees C groups, and this inhibition persisted for the duration of the experiment. There was no statistically significant difference in the results between the storage temperatures. CONCLUSIONS: These data suggest that MMC retains its antiproliferative effect for at least 6 weeks following reconstitution and that this effect is not changed by storage temperature. Cost savings may be realized by storing MMC in unit-dose reconstituted aliquots for these longer periods, providing sterility can be assured.

ENVIRONMENTAL AUDITING: An Approach for Characterizing Tropospheric Ozone Risk to Forests

/ The risk tropospheric ozone poses to forests in the United States is dependent on the variation in ozone exposure across the distribution of the forests in question and the various environmental and climate factors predominant in the region. All these factors have a spatial nature, and consequently an approach to characterization of ozone risk is presented that places ozone exposure-response functions for species as seedlings and model-simulated tree and stand responses in a spatial context using a geographical information systems (GIS). The GIS is used to aggregate factors considered important in a risk characterization, including: (1) estimated ozone exposures over forested regions, (2) measures of ozone effects on species' and stand growth, and (3) spatially distributed environmental, genetic, and exposure influences on species' response to ozone. The GIS-based risk characterization provides an estimation of the extent and magnitude of the potential ozone impact on forests. A preliminary risk characterization demonstrating this approach considered only the eastern United States and only the limited empirical data quantifying the effect of ozone exposures on forest tree species as seedlings. The area-weighted response of the annual seedling biomass loss formed the basis for a sensitivity ranking: sensitive-aspen and black cherry (14%-33% biomass loss over 50% of their distribution); moderately sensitive-tulip popular, loblolly pine, eastern white pine, and sugar maple (5%-13% biomass loss); insensitive-Virginia pine and red maple (0%-1% loss). In the future, the GIS-based risk characterization will include process-based model simulations of the three- to 5-year growth response of individual species as large trees with relevant environmental interactions and model simulated response of mixed stands. The interactive nature of GIS provides a tool to explore consequences of the range of climate conditions across a species' distribution, forest management practices, changing ozone precursors, regulatory control strategies, and other factors influencing the spatial distribution of ozone over time as more information becomes available.KEY WORDS: Ecological risk assessment; GIS; Ozone; Risk characterization; Forests; Trees

Effects of cytoskeleton-reactive agents on aqueous outflow facility in a porcine ocular anterior segment preparation.

OBJECTIVE: To assess the effect of three agents known to react with cytoskeletal components--acrylamide (a vinyl monomer), demecolcine (a colchicine derivative) and stypoldione (an ichthyotoxic metabolite derived from the tropical marine alga Stypopodium zonale)--on facility of aqueous humour outflow in order to increase the understanding of the role of the cytoskeleton in the biology of aqueous outflow. DESIGN: Constant-pressure perfusion experiments with various concentrations of the agents using a porcine ocular anterior segment preparation. NUMBERS: Eighteen experimental eyes (six with each agent) and 18 control eyes. MEAN OUTCOME MEASURES: Aqueous outflow facility, determined after 1, 2 and 24 hours of perfusion. RESULTS: The mean baseline outflow facility was 0.10 (standard error of the mean 0.045) microL/min per mm Hg. There was no significant difference in mean baseline outflow facility between the experimental and control eyes. Perfusion of 0.01 to 1.0 mM demecolcine did not produce any consistent alteration in outflow facility. A mild to moderate dose-related experimental effect was noted after 2 hours of perfusion with 1.0 to 100 mM acrylamide. Perfusion with 0.10 mM stypoldione produced a moderate increase in outflow facility; the effect was greatest after 1 hour of perfusion. CONCLUSIONS: Our preliminary results do not support a central role for the cytoskeleton in the regulation of aqueous outflow resistance.

Increased prevalence of occludable angles and angle-closure glaucoma in patients with pseudoexfoliation.

The association between angle-closure glaucoma and pseudoexfoliation is controversial. We retrospectively studied the angle configurations of 54 patients with pseudoexfoliation and found gonioscopically occludable angles in five cases (9.3%). We also analyzed the data from several large studies of pseudoexfoliation patients and found an increased prevalence of acute angle-closure glaucoma in this group. These data suggest that pseudoexfoliation patients may represent a high-risk population for the development of angle-closure glaucoma.

The effect of intracameral ethacrynic acid on the intraocular pressure of living monkeys.

Previous studies have shown that the sulfhydryl-reactive ethacrynic acid increases outflow facility in living monkeys when perfused via the anterior chamber. To study its potential clinical use further, living monkeys were intracamerally injected with 10 microliters of ethacrynic acid, with concentrations ranging from 0.5 to 7.5 mmol/l. The fellow control eye was injected with 10 microliters of diluent. The status of the anterior segment was monitored by slit-lamp biomicroscopy and the intraocular pressure was measured by pneumatonometry with the monkeys anesthetized with ketamine. The anterior segment of living monkeys tolerated injections up to 3.0-mmol/l ethacrynic acid without marked adverse effects. One of 13 monkey eyes injected with 3.0-mmol/l ethacrynic acid demonstrated mild reversible segmental corneal edema. The greatest mean intraocular pressure reduction in the 3.0- to 3.75-mmol/l group occurred at six hours, with the experimental intraocular pressure decreasing 2.9 mm Hg compared to a mean intraocular pressure increase of 0.1 mm Hg in the control group (n = 19). Concentrations of ethacrynic acid less than 3.0 mmol/l did not provide reliable reduction of intraocular pressure, whereas concentrations greater than 3.75 mmol/l caused a greater incidence and severity of corneal edema. We believe that the intracameral injection of ethacrynic acid can reliably and safely reduce intraocular pressure in living monkey eyes, and that this drug deserves further investigation as a potential antiglaucomatous agent.

Effects of topical ethacrynic acid adducts on intraocular pressure in rabbits and monkeys.

We evaluated the effect of topical ethacrynic acid on rabbit and monkey intraocular pressure. In a preliminary experiment, 100-mmol/L ethacrynic acid applied topically to Dutch-Belted rabbit eyes was associated with an 8-mm Hg lowering of intraocular pressure. However, corneal edema was severe, and the corneal epithelium sloughed off. To try to maintain the pressure-lowering effect but reduce the corneal side effects, we attempted to create an adduct of ethacrynic acid by utilizing ethacrynic acid's sulfhydryl reactivity. Ethacrynic acid was mixed with equimolar cysteine to bind the sulfhydryl-reactive sites on ethacrynic acid. The goal was to expose the cornea to adducted ethacrynic acid, which might then dissociate in the anterior chamber via a retro-Michael reaction. Intraocular pressure decreased 8.9 mm Hg (n = 40) with this treatment, and corneal edema was lessened (32 of 40 eyes had mild to no edema). However, we observed that when the eye was treated before ethacrynic acid-cysteine administration with topical acetylcysteine, the corneal side effects were reduced further and the intraocular pressure effect remained. In living cynomolgus monkeys receiving a single pretreatment drop of 75-mmol/L acetylcysteine followed by two drops of 130-mmol/L ethacrynic acid and 130-mmol/L cysteine, an intraocular pressure lowering of 9.9 mm Hg was observed (n = 7). However, in three of seven eyes corneal edema developed. Pretreatment with two drops of acetylcysteine eliminated the pressure-lowering effect but did not confer any added corneal protection. Our results indicate that topical ethacrynic acid-cysteine is effective in lowering the intraocular pressure of rabbits and cynomolgus monkeys and, when combined with acetylcysteine pretreatment, may offer the potential for a new topical therapeutic regimen for use in glaucoma.