Search

The eruption of the Hunga Tonga-Hunga Ha'apai volcano on 15 January 2022 offered a good opportunity to explore the early impacts of tropical volcanic eruptions on stratospheric composition. Balloon-borne observations near Réunion Island revealed the unprecedented amount of water vapor injected by the volcano. The enhanced stratospheric humidity, radiative cooling, and expanded aerosol surface area in the volcanic plume created the ideal conditions for swift ozone depletion of 5% in the tropical stratosphere in just 1 week. The decrease in hydrogen chloride by 0.4 parts per million by volume (ppbv) and the increase in chlorine monoxide by 0.4 ppbv provided compelling evidence for chlorine activation within the volcanic plume. This study enhances our understanding of the effect of this unusual volcanic eruption on stratospheric chemistry and provides insights into possible chemistry changes that may occur in a changing climate.

As part of the UV-Indien project, a station for measuring ultraviolet radiation and the cloud fraction was installed in December 2019 in Moroni, the capital of the Comoros, situated on the west coast of the island of Ngazidja. A ground measurement campaign was also carried out on 12 January 2020 during the ascent of Mount Karthala, located in the center of the island of Ngazidja. In addition, satellite estimates (Ozone Monitoring Instrument and TROPOspheric Monitoring Instrument) and model outputs (Copernicus Atmospheric Monitoring Service and Tropospheric Ultraviolet Model) were combined for this same region. On the one hand, these different measurements and estimates make it possible to quantify, evaluate, and monitor the health risk linked to exposure to ultraviolet radiation in this region, and, on the other, they help to understand how cloud cover influences the variability of UV-radiation on the ground. The measurements of the Ozone Monitoring Instrument onboard the EOS-AURA satellite, being the longest timeseries of ultraviolet measurements available in this region, make it possible to quantify the meteorological conditions in Moroni and to show that more than 80% of the ultraviolet indices are classified as high and that 60% of these are classified as extreme. The cloud cover measured in Moroni by an All Sky Camera was used to distinguish between the cases of UV index measurements taken under clear or cloudy sky conditions. The ground-based measurements thus made it possible to describe the variability of the diurnal cycle of the UV index and the influence of cloud cover on this parameter. They also permitted the satellite measurements and the results of the simulations to be validated. In clear sky conditions, a relative difference of between 6 and 11% was obtained between satellite or model estimates and ground measurements. The ultraviolet index measurement campaign on Mount Karthala showed maximum one-minute standard erythemal doses at 0.3 SED and very high daily cumulative erythemal doses at more than 80 SED. These very high levels are also observed throughout the year and all skin phototypes can exceed the daily erythemal dose threshold at more than 20 SED.

Reunion Island is a popular tourist destination with sandy beaches, an active volcano (Piton de la Fournaise), and Piton des Neiges, the highest and most dominant geological feature on the island. Reunion is known to have high levels of solar ultraviolet radiation (UVR) with an ultraviolet index (UVI) which can reach 8 in winter and 16 in summer (climatological conditions). UVR has been linked to skin cancer, melanoma, and eye disease such as cataracts. The World Health Organization (WHO) devised the UVI as a tool for expressing UVR intensity. Thresholds ranging from low (UVI 1-2) to extreme (UVI > 11) were defined depending on the risk to human health. The purpose of the study was to assess UVR exposure levels over three of the busiest tourist sites on the island. UVR was measured over several hours along popular hiking trails around Piton de la Fournaise (PDF), Piton des Neiges (PDN), and St-Leu Beach (LEU). The results were compared with those recorded by the local UV station at Saint-Denis. In addition, cumulative standard erythemal dose (SED) was calculated. Results showed that UVI exposure at PDF, PDN, and LEU were extreme (>11) and reached maximum UVI levels of 21.1, 22.5, and 14.5, respectively. Cumulative SEDs were multiple times higher than the thresholds established by the Fitzpatrick skin phototype classification. UVI measurements at the three study sites showed that Reunion Island is exposed to extreme UVR conditions. Public awareness campaigns are needed to inform the population of the health risks related to UVR exposure.

Measurement of solar ultraviolet radiation (UVR) is important for the assessment of potential beneficial and adverse impacts on the biosphere, plants, animals, and humans. Excess solar UVR exposure in humans is associated with skin carcinogenesis and immunosuppression. Several factors influence solar UVR at the Earth's surface, such as latitude and cloud cover. Given the potential risks from solar UVR there is a need to measure solar UVR at different locations using effective instrumentation. Various instruments are available to measure solar UVR, but some are expensive and others are not portable, both restrictive variables for exposure assessments. Here, we compared solar UVR sensors commercialized at low or moderate cost to assess their performance and quality of measurements against a high-grade Bentham spectrometer. The inter-comparison campaign took place between March 2018 and February 2019 at Saint-Denis, La Réunion. Instruments evaluated included a Kipp&Zonen UVS-E-T radiometer, a Solar Light UV-Biometer, a SGLux UV-Cosine radiometer, and a Davis radiometer. Cloud fraction was considered using a SkyCamVision all-sky camera and the Tropospheric Ultraviolet Visible radiative transfer model was used to model clear-sky conditions. Overall, there was good reliability between the instruments over time, except for the Davis radiometer, which showed dependence on solar zenith angle. The Solar Light UV-Biometer and the Kipp&Zonen radiometer gave satisfactory results, while the low-cost SGLux radiometer performed better in clear sky conditions. Future studies should investigate temporal drift and stability over time.

Predicting and understanding the biological response to future climate change is a pressing challenge for humanity. In the 21st century, many species will move into higher latitudes and higher elevations as the climate warms. In addition, the relative abundances of species within local assemblages are likely to change. Both effects have implications for how ecosystems function. Few biodiversity forecasts, however, take account of both shifting ranges and changing abundances. We provide a novel analysis predicting the potential changes to assemblage-level relative abundances in the 21st century. We use an established relationship linking ant abundance and their colour and size traits to temperature and UV-B to predict future abundance changes. We also predict future temperature driven range shifts and use these to alter the available species pool for our trait-mediated abundance predictions. We do this across three continents under a low greenhouse gas emissions scenario (RCP2.6) and a business-as-usual scenario (RCP8.5). Under RCP2.6, predicted changes to ant assemblages by 2100 are moderate. On average, species richness will increase by 26%, while species composition and relative abundance structure will be 26% and 30% different, respectively, compared with modern assemblages. Under RCP8.5, however, highland assemblages face almost a tripling of species richness and compositional and relative abundance changes of 66% and 77%. Critically, we predict that future assemblages could be reorganized in terms of which species are common and which are rare: future highland assemblages will not simply comprise upslope shifts of modern lowland assemblages. These forecasts reveal the potential for radical change to montane ant assemblages by the end of the 21st century if temperature increases continue. Our results highlight the importance of incorporating trait-environment relationships into future biodiversity predictions. Looking forward, the major challenge is to understand how ecosystem processes will respond to compositional and relative abundance changes., (© 2019 John Wiley & Sons Ltd.)

We have derived values of the Ultraviolet Index (UVI) at solar noon using the Tropospheric Ultraviolet Model (TUV) driven by ozone, temperature and aerosol fields from climate simulations of the first phase of the Chemistry-Climate Model Initiative (CCMI-1). Since clouds remain one of the largest uncertainties in climate projections, we simulated only the clear-sky UVI. We compared the modelled UVI climatologies against present-day climatological values of UVI derived from both satellite data (the OMI-Aura OMUVBd product) and ground-based measurements (from the NDACC network). Depending on the region, relative differences between the UVI obtained from CCMI/TUV calculations and the ground-based measurements ranged between -5.9% and 10.6%. We then calculated the UVI evolution throughout the 21st century for the four Representative Concentration Pathways (RCPs 2.6, 4.5, 6.0 and 8.5). Compared to 1960s values, we found an average increase in the UVI in 2100 (of 2-4%) in the tropical belt (30°N-30°S). For the mid-latitudes, we observed a 1.8 to 3.4 % increase in the Southern Hemisphere for RCP 2.6, 4.5 and 6.0, and found a 2.3% decrease in RCP 8.5. Higher increases in UVI are projected in the Northern Hemisphere except for RCP 8.5. At high latitudes, ozone recovery is well identified and induces a complete return of mean UVI levels to 1960 values for RCP 8.5 in the Southern Hemisphere. In the Northern Hemisphere, UVI levels in 2100 are higher by 0.5 to 5.5% for RCP 2.6, 4.5 and 6.0 and they are lower by 7.9% for RCP 8.5. We analysed the impacts of greenhouse gases (GHGs) and ozone-depleting substances (ODSs) on UVI from 1960 by comparing CCMI sensitivity simulations (1960-2100) with fixed GHGs or ODSs at their respective 1960 levels. As expected with ODS fixed at their 1960 levels, there is no large decrease in ozone levels and consequently no sudden increase in UVI levels. With fixed GHG, we observed a delayed return of ozone to 1960 values, with a corresponding pattern of change observed on UVI, and looking at the UVI difference between 2090s values and 1960s values, we found an 8 % increase in the tropical belt during the summer of each hemisphere. Finally we show that, while in the Southern Hemisphere the UVI is mainly driven by total ozone column, in the Northern Hemisphere both total ozone column and aerosol optical depth drive UVI levels, with aerosol optical depth having twice as much influence on the UVI as total ozone column does.

South Africa has been measuring the ground-based solar UV index for more than two decades at six sites to raise awareness about the impacts of the solar UV index on human health. This paper is an exploratory study based on comparison with satellite UV index measurements from the OMI/AURA experiment. Relative UV index differences between ground-based and satellite-derived data ranged from 0 to 45% depending on the site and year. Most of time, these differences appear in winter. Some ground-based stations' data had closer agreement with satellite-derived data. While the ground-based instruments are not intended for long-term trend analysis, they provide UV index information for public awareness instead, with some weak signs suggesting such long-term trends may exist in the ground-based data. The annual cycle, altitude, and latitude effects clearly appear in the UV index data measured in South Africa. This variability must be taken into account for the development of an excess solar UV exposure prevention strategy., Competing Interests: The authors declare no conflict of interest.

Phocine distemper virus (PDV) was first recognized in 1988 following a massive epidemic in harbor and grey seals in north-western Europe. Since then, the epidemiology of infection in North Atlantic and Arctic pinnipeds has been investigated. In the western North Atlantic endemic infection in harp and grey seals predates the European epidemic, with relatively small, localized mortality events occurring primarily in harbor seals. By contrast, PDV seems not to have become established in European harbor seals following the 1988 epidemic and a second event of similar magnitude and extent occurred in 2002. PDV is a distinct species within the Morbillivirus genus with minor sequence variation between outbreaks over time. There is now mounting evidence of PDV-like viruses in the North Pacific/Western Arctic with serological and molecular evidence of infection in pinnipeds and sea otters. However, despite the absence of associated mortality in the region, there is concern that the virus may infect the large Pacific harbor seal and northern elephant seal populations or the endangered Hawaiian monk seals. Here, we review the current state of knowledge on PDV with particular focus on developments in diagnostics, pathogenesis, immune response, vaccine development, phylogenetics and modeling over the past 20 years.

The physiologic mechanism that underlies the epinephrine-induced increase in facility of outflow (C) in glaucomatous human eyes and normal primate eyes is not completely understood. In this study, a recently developed in vitro human eye perfusion model was used to simultaneously monitor facility and cyclic adenosine monophosphate (AMP) changes in response to epinephrine (EPI). In this system, EPI (2.5 x 10(-5) mol/l) resulted in a maximal 44% increase in C, with an ED50 occurring at approximately 8 x 10(-6) mol/l. The C-increasing effect of 10(-5) mol/l EPI was unaffected by 10(-6) mol/l phentolamine. However, it was completely blocked in the presence of 10(-6) mol/l timolol or 2 x 10(-7) mol/l ICI118,551, suggesting the involvement of beta-2 adrenergic receptors. In biochemical studies, 10(-5) mol/l EPI induced a 12- to 14-fold increase in cyclic AMP in the perfusate of treated eyes. This increase was blocked by ICI118,551. In isolated intact human trabecular tissue, a 10 min incubation with 10(-5) EPI stimulated cyclic AMP by a factor of 2.7 over control levels. After 90 min, cyclic AMP levels were increased 4.2 fold over control levels. Collectively, these results show that the intraocular pressure lowering effect of EPI in the human eye is mediated, at least in part, by an increase in facility of outflow. Furthermore, the facility increase appears to be mediated by beta-2 adrenergic receptors and is correlated in time with increased cyclic AMP production.

Heat-soaked PCR (HS-PCR) is a method for enhancing amplification performed by heating the DNA sample at 94 degrees C in 90 microliters 1.1 x buffer for 30 min. A 10-microliters bolus of concentrated (10x) deoxynucleotides, Taq DNA polymerase and primers prepared without buffer is then added just prior to thermal cycling. We have investigated the application of this method in a variety of forensically important DNA samples and compared it with regular PCR (R-PCR). DNA samples extracted from bone, postmortem tissues, bloodstains and hair contained low concentrations of human DNA or were contaminated with either non- human DNA or hemoglobin degradation products. Optimal conditions for HS-PCR were determined for the 3' ApoB VNTR locus and applied to a centromeric repeat element and to a single-copy locus. HS-PCR consistently and reproducibly enhanced product yield and specificity over R-PCR at all three loci in the entire set of DNA samples. HS-PCR was also effective in overcoming the inhibitory effect of hemoglobin at concentrations that fully impeded R-PCR.

Cell cultures derived from trabecular meshworks of human and bovine eyes and from bovine vascular endothelia were incubated at 37 degrees C for 1 hr with ethacrynic acid (ECA, 0.1-0.5 mmol/l) dissolved in culture medium. At 2 hr after the initial exposure, ECA at concentrations up to 0.4 mmol/l induced a reversible alteration in cell shape in all three cell types that was coincident with a change in the staining pattern of major cytoskeletal components including actin, alpha-actinin, vinculin, and vimentin. Distinct progressive alterations in beta-tubulin also occurred, with initial changes observed 10 min after ECA exposure. The ECA-induced changes in tubulin were blocked in part by preincubation with taxol (which stabilizes the microtubule structure), but they appeared to differ from those occurring with nocodazole (which interferes with tubulin assembly). These results suggest the possibility that ECA-induced increases in outflow facility may be mediated by alterations in the cytoskeletons of outflow pathway cells.

Anterior segments of human donor eyes were perfused with culture medium at a perfusion pressure of 15 mm Hg in a 5% carbon dioxide environment at 37 degrees C. After determination of a baseline facility of outflow, the perfusion chamber contents were exchanged with either drug vehicle or ethacrynic acid, at concentrations ranging from 0.01 to 0.25 mmol/L, after which postdrug facility was measured in the continuous presence of drug vehicle or ethacrynic acid. Ethacrynic acid increased facility of outflow from 28% to 105% at ethacrynic acid concentrations of 0.01 to 0.25 mmol/L, respectively. No morphologic correlate of the facility increase was observed with 0.01-mmol/L ethacrynic acid, nor were there any signs of cellular toxic effects. At 0.1 mmol/L, separations between trabecular meshwork cells and breaks between inner-wall cells were observed. At 0.25 mmol/L, focal areas of cell swelling and necrosis were noted. This study demonstrated that ethacrynic acid increases outflow facility in the aged human eye at concentrations that produce no apparent toxic effects. Therefore, ethacrynic acid may potentially prove useful in the treatment of glaucoma.

Twice daily topical administration of echothiophate for 2 weeks to the eyes of living cynomolgus monkeys produced profound subsensitivity of the accommodative response to pilocarpine and an approximately 50% decrease in the number of specific binding sites for 3H-quinuclidinyl benzilate (3H-QNB) in the ciliary muscle without a change in their affinity. When echothiophate treatment was discontinued, functional cholinergic sensitivity and the number of QNB binding sites both returned to normal over a similar 4-8 week period. Most animals had a modest overshoot of both functional sensitivity and number of binding sites for at least several weeks thereafter. The treated to control eye ratios for the number of binding sites and accommodative response to pilocarpine were correlated and the plot of log treated to control binding site ratio versus treated to control accommodation ratio resembled a dose-response curve. Similarly, the treated versus control eye differences for the two parameters were correlated, with the regression line passing through the 0-0 axis intercept. Collectively, these findings suggest that agonist-induced modulation of functional cholinergic sensitivity in the parasympathetically innervated (as opposed to denervated) ciliary muscle occurs by a muscarinic receptor-mediated mechanism. This system appears to provide a useful model to study the regulation of ciliary muscle cholinergic sensitivity.

We have recently developed a tissue model of the human aqueous outflow pathway involving placement of the eviscerated anterior corneoscleral shell, [with lens and uveal tissue removed but trabecular meshwork (TM) attached] onto a specialized perfusion apparatus. The TM and associated outflow tissues are perfused with culture medium at a physiologically-relevant perfusion pressure in a 5% CO2 environment at 37 degrees C. Under these conditions, the perfused outflow tissues are similar for several days, to the human and/or subhuman primate outflow system in vivo with regard to morphology as well as several functional parameters. Measured facility of outflow (0.271 +/- 0.018 microliters min-1 mmHg-1, n = 79) is similar to facility values obtained by tonography in living human beings. Moreover, outflow facility decreases in a linear fashion with increased perfusion pressure by 1.4% mmHg-1. Finally the removal of the TM results in a 41% decrease in measured outflow resistance. The ability to study viable human outflow tissue for at least several days and the opportunity to establish a model which serves as an alternative to animal testing, point to the potential importance of this technique in investigating the biology of the aqueous outflow system.

The effect of the vasoactive peptide, endothelin, on facility of outflow, accommodation, and pupil diameter was measured in the monkey eye in vivo. Endothelin increased the outflow facility 22-71% at approximate anterior chamber concentrations ranging from 10(-10)-10(-7) M. Endothelin-induced accommodation was modest but consistent and statistically significant ranging from 1.61 to a maximum of 2.43 diopters at 10(-9) M and 10(-7) M endothelin, respectively. No change in pupil diameter was noted with any of the administered doses. These data demonstrate an action of endothelin on outflow facility and, together with prior evidence indicating the presence of endothelin receptors on the ciliary muscle, suggest that, like cholinergic agonists, the observed effects of endothelin on outflow facility may be mediated, at least in part, through an action on the ciliary muscle.

During the course of constant-pressure anterior chamber perfusion experiments in eyes from a number of species, measured outflow facility increases progressively with perfusion time. One possible explanation for this phenomenon has been that the technique of perfusion induces a loss of extracellular material from the outflow pathway. Therefore, this general observation has been termed the "wash-out" effect. Data from the authors' laboratories demonstrate that although outflow facility increases in cynomolgus monkey eyes by 26% and 42% from baseline over 90 minutes in vivo and in vitro, respectively, this does not occur in the enucleated human eye. The absence of washout in the human eye probably cannot be explained by postmortem conditions since enucleated monkey eyes undergo a similar magnitude of washout as the monkey eye in vivo. Furthermore, since washout does not occur in the perfused human infant eye, the age of the donor cannot explain the difference in washout properties between human and other primate eyes. The monkey eye in vivo is a valuable model in the study of outflow physiology. However, the difference in washout behavior between human and other primate eyes may point to a potentially important fundamental difference in the biology of the outflow pathways in these two species.

A smooth muscle cell line (H7CM) was established from the ciliary muscle of a 1-day-old human infant. The cultured cells had a normal female karyotype (46 XX) and could be maintained in cell culture for at least 11 generations. A common feature of confluent cultures was the presence of abundant bundles of 6-7 nm microfilaments associated with dense bodies. Both the ultrastructural appearance and the presence of smooth muscle-specific alpha-isoactin (also present in the human ciliary muscle in situ) support the smooth muscle origin of the H7CM cell line. Continuous membrane voltage (Vm) recordings were obtained in confluent monolayers of H7CM cells using glass microelectrodes. Resting Vm in 105 impalements averaged -66.2 +/- 0.7 mV (mean +/- standard error of the mean). In this system, rapid membrane transients induced by changing of the superfusing test solutions were detectable. Relative K+ conductance was characterized, and the contribution of electrogenic sodium/potassium adenosine triphosphatase to Vm was investigated. Under control conditions, H7CM cells were electrically quiescent. However, action potentials could be induced by application of 10 mM barium. Barium-induced action potentials were not abolished by removal of extracellular Na+ nor were they inhibited by the presence of tetrodotoxin. However, they were blocked by verapamil, fulfilling criteria believed to be typical for smooth muscle cells. Acetylcholine, carbachol, and to a lesser extent pilocarpine induced a reversible Vm depolarization. The effect of acetylcholine was blocked by atropine, implying muscarinic receptor involvement in the Vm response. Collectively, these findings show the potential usefulness of cultured ciliary muscle cells in understanding further the cellular mechanisms underlying drug-induced contraction of the human ciliary muscle.

Choline acetyltransferase (ChAT; EC 2.3.1.6) and acetylcholinesterase (AChE; EC 3.1.1.7) activities were measured in cynomolgus monkey ciliary muscle 1 month and 6 or more months after ciliary ganglionectomy (CG) or post-ganglionic ciliary neurectomy (PCN). ChAT activity was undetectable and AChE activity was elevated 1 month after CG or PCN, while both averaged about 30% of normal levels 6 or more months after denervation. Four out of six eyes reinnervated by functional criteria 6-12 months after CG or PCN. In one of the two remaining eyes permanently denervated, ChAT was absent from the ciliary muscle. In the other, ChAT activity was about 50% of normal, similar to the reinnervated eyes, but the regenerated cholinergic nerves were not approximated to the ciliary muscle fibers.

The dose/effect relationships for intracameral doses of aceclidine HCl and pilocarpine HCl on total facility of outflow and accommodative amplitude were compared in the cynomolgus monkey eye in vivo. The largest dose of aceclidine tested (20 micrograms), resulted in an average increase of 270% in outflow facility, whereas accommodation was limited to an average of 5 diopters. In contrast, 20 micrograms of intracameral pilocarpine (which is submaximal in increasing facility) resulted in an average of 19 diopters of accommodation. These results provide further evidence for a dissociation between the accommodative and outflow facility functions of the ciliary muscle.

Ciliary ganglionectomy in the cynomolgus monkey produced loss of the accommodative response to electrical stimulation of the Edinger-Westphal nucleus and to topical eserine, concurrent with enhanced responsiveness to topical and systemic pilocarpine. This supersensitivity produced by parasympathetic denervation was not associated with an increase in muscarinic receptor number or affinity measured using specific QNB binding sites in ciliary muscle homogenates. In seven of ten cases, ciliary muscle muscarinic receptors were decreased by 60-84%, while in the remaining three cases, the decrease ranged from 12-37%. In no case was supersensitivity accompanied by an increased number of receptors. Six or more months after ganglionectomy, accommodative responses to central electrical stimulation and topical eserine returned to normal in most animals, indicating parasympathetic reinnervation of the ciliary muscle. Ciliary muscle sensitivity to pilocarpine returned to normal with reinnervation and was associated with recovery of normal receptor number and with normal binding affinity.

In cynomolgus monkeys, resting total outflow facility was unaltered 1 and 6 or more months after ciliary ganglionectomy (CG) or postganglionic ciliary neurectomy (PCN). Intraocular pressure (IOP) was decreased in the denervated eye 1 week and 1 month after surgery, but returned to normal after 6 or more months. Although baseline facilities were comparable in CG/PCN and fellow control eyes 6 or more months after surgery, even maximal intracameral doses of pilocarpine did not increase outflow facility in previously denervated eyes, while a normal facility increase occurred in fellow control eyes. However, both previously denervated and fellow control eyes exhibited a large facility increase to both submaximal and greater than maximal intracameral doses of eserine.

Topical echothiophate administration to the cynomolgus monkey eye for 5-6.5 months produced marked subsensitivity of the accommodative response to pilocarpine and a 65% decrease in specific high affinity 3H-QNB binding sites (ostensibly indicating muscarinic receptors) in the ciliary muscle. The decrease in 3H-QNB binding sites was quantitatively similar in surgically untouched, totally iridectomized, and ciliary muscle disinserted eyes. Following a 5-month off treatment recovery period, 3H-QNB binding sites increased to more than twice the number in untreated control eyes. In echothiophate-treated eyes whose contralateral eyes had previously received atropine+echothiophate, 3H-QNB binding sites were three to six times more numerous than in other long-term echothiophate-treated eyes, and one to two times more numerous than in untreated controls. Topical pilocarpine administration for 1 day to 7 months reduced ciliary muscle 3H-QNB binding sites by approximately 25%. Alterations in muscarinic receptors during and following cholinergic drug therapy may in part explain the observed subsensitization and recovery of ciliary muscle physiological responses.

We have recently developed a technique for constant pressure perfusion of the aqueous outflow pathway of the eye. Our preliminary studies, conducted in the calf eye, show surprisingly that the manipulations necessary for preparing the outflow pathways and attached corneoscleral shell for perfusion do not greatly disrupt normal aqueous outflow physiology and anatomy according to the following criteria: 1. facility of outflow is similar before and during outflow pathway perfusion 2. as in the intact eye, facility of outflow decreases with increased IOP 3. removal of outflow resistance tissue greatly increases facility of outflow 4. morphology of outflow tissues remains normal Use of the perfused outflow pathway model may enable the creation of valuable in vitro preparations which may provide much needed information about the pathogenesis of primary open-angle glaucoma.

The ciliary muscle which is involved in accommodation and regulation of aqueous humour outflow resistance resembles smooth muscle in other parts of the body. In the present investigation we used an established primary cell line (H7CM) to study the effects of endothelin, a novel vasoconstrictor peptide, on membrane voltage (V) and intracellular calcium in cultured human ciliary muscle cells. Membrane voltage was measured in confluent monolayers of H7CM cells using conventional microelectrodes. Intracellular calcium concentration [( Ca]i) was measured in single H7CM cells using the fluorescent calcium indicator fura-2. Under resting conditions V averaged -66.9 +/- 0.7 mV (mean +/- SEM, n = 125). Endothelin (10(-10)-10(-6)M) induced a dose-dependent reversible membrane voltage depolarization and a dose-dependent rise in [Ca]i. The initial calcium peak was followed by a recovery phase during which oscillations of [Ca]i occurred. The initial calcium peak was not dependent on the presence of extracellular calcium and was not abolished in the presence of the calcium antagonist verapamil (10(-4)M). Thus it is probably mediated by a release of calcium from intracellular reservoirs. We conclude that cultured human ciliary muscle cells express a functional endothelin receptor.

Surgically virgin (N), aniridic (I), and ciliary muscle disinserted (D) cynomolgus monkey eyes had total outflow facility determined in vivo for 88-136 minutes by two-level constant pressure perfusion of the anterior chamber. In all three types of eyes, facility increased during the first hour but stabilized thereafter. Average facility for the second half hour was 15% higher than for the first half hour in N and I eyes, and 10% higher in D eyes. Altered iris muscle tone can therefore not explain the facility increase, while increased ciliary muscle tone might explain part but not all of it. Judicious use of the data may aid interpretation of perfusion experiments conducted under similar conditions.

Ciliary ganglionectomy and/or postganglionic ciliary neurectomy in the cynomolgus monkey was followed by supersensitivity to intramuscular (i.m.) pilocarpine and lack of response to topical eserine and to electrical stimulation of the Edinger-Westphal nucleus. Normal responsiveness to pilocarpine and eserine returned in most instances after about 6 months. An accommodative response to stimulus of the Edinger-Westphal nucleus was also present and, as in control eyes, could be blocked by hexamethonium. Collectively, these findings demonstrate that parasympathetic innervation to the ciliary muscle mediating accommodation traverses a typical peripheral autonomic synapse, almost certainly located predominantly in the ciliary ganglion; by 6 months after denervation, the ciliary muscles have reinnervated; and the parasympathetic pathway to the eye exhibits plasticity and capacity for regeneration.

No differences were found between fluorophotometrically determined aqueous flow rates in cynomolgus monkeys having vs not having undergone prior repeated anterior chamber perfusion. This suggests that: repeated perfusion does not produce a progressive decline in the rate of aqueous formation; and the repeatedly perfused monkey eye can be validly used in studies of aqueous formation.