Adenylyl and guanylyl cyclase activity in the choroid.

The choroid, a low-resistance vascular structure provides nourishment to and removal of potential toxic waste products from the adjacent non-vascularized outer layers of the retina, macula, and optic disc region and may be involved in the pathology of normal tension glaucoma. This study is aimed at delineating the biochemical pharmacology of vascular tone control in this tissue. By using a procedure to harvest fresh choroidal tissue, we studied some basic characteristics of the adenylyl (AC) and guanylyl cyclase (GC) enzyme systems in this tissue using the 3H-cAMP/32P-ATP tracer method. Compared to respective baseline measurements (100+/-SEM%), AC activity was stimulated (p < 0.05) by forskolin (FSK, 477+/-59%), fluoroaluminate (AlF(4), 360+/-10.3%), isoproterenol (ISO, 129+/-5.5%), vasoactive intestinal peptide (VIP, 132+/-6.1%), calmodulin (CAM)+Mn2+ (196+/-69%), and dose-dependently by prostaglandin (PG) E2 (up to 162+/-3.6%). The antagonist drug calmidazolium inhibited the CAM-dependent increase but also blocked basal activity (47+/-2.0%) without affecting the FSK response. Other CAM blockers (TFP, W5) produced similar results but were not completely selective for CAM-activated cyclase. GDPbetaS, a blocker of G-protein coupling to AC did not affect AC responses to FSK, ISO, and AlF4, but decreased the response to PGE2. N-ethyl-carboxamido adenosine (NECA), which activates adenosine A2 receptors, did not synergize with FSK or add to ISO responses and did not activate AC by itself. In the GC system activity was stimulated (p< 0.05) by CAM+Mn2+ (239+/-27%), by atrial natriuretic peptide (ANP, up to 143+/-1.4%) and sodium nitroprusside (SNP, up to 179+/-1.6%). These results show that choroidal tissue has significant activities of the adenylyl and guanylyl cyclase second messenger systems potentially responsive to hormones/neurotransmitters that may control the degree of relaxation in this vascular tissue.

Effect of pilocarpine 4% in combination with latanoprost 0.005% or 8-iso prostaglandin E2 0.1% on intraocular pressure in laser-induced glaucomatous monkey eyes.

PURPOSE: To compare the effect of pilocarpine, an agent that reduces uveoscleral outflow, on the ocular hypotensive efficacy of latanoprost and 8-iso prostaglandin E2 (PGE2). METHODS: Each of the two treatment groups was composed of the same eight monkeys with unilateral laser-induced glaucoma. Intraocular pressure (IOP) was measured hourly for 6 hours beginning at 9:00 AM on the baseline day (Thursday before treatment week) and on treatment days 1, 3, and 5 (Monday, Wednesday, and Friday). On all five treatment days, one drop of pilocarpine 4% was administered at 9:00 AM and 3:00 PM and one drop of latanoprost 0.005% or 25 microL of 8-iso PGE2 0.1% was administered at 10:00 AM and 4:00 PM. RESULTS: One hour after pilocarpine instillation on day 1, the reduction of IOP was similar (P > 0.90) in both treatment groups, 7.6 +/- 1.1 mm Hg (mean +/- standard error of the mean ) in the latanoprost group and 7.4 +/- 0.8 mm Hg in the 8-iso PGE2 group. However, the IOP effects of the two treatment groups became significantly different (P < 0.05) beginning 2 hours after dosing with latanoprost or 8-iso PGE, on day 1. A difference (P < 0.05) between the two groups persisted at all subsequent measurements. The reduction of IOP lessened with repeated dosing in the latanoprost and 8-iso PGE2 groups. Three hours after dosing with pilocarpine and two hours after dosing with the prostanoids, the IOP reduction was 8.3 +/- 0.9 mm Hg in the latanoprost group and 9.9 +/- 0.6 mm Hg in the 8-iso PGE2 group on day 1, and 2.1 +/- 1.0 mm Hg in the latanoprost group and 7.3 +/- 0.9 mm Hg in the 8-iso PGE1 group on day 5. CONCLUSIONS: The smaller reductions in IOP with pilocarpine and latanoprost than with pilocarpine and 8-iso PGE2 show that pilocarpine blocks much more of the ocular hypotensive effect of latanoprost than of 8-iso PGE2. The results also indicate that pilocarpine and latanoprost are mutually antagonistic. Enhancement of uveoscleral outflow appears to account for most of the ocular hypotensive effect of latanoprost and for much less of the ocular hypotensive effect of 8-iso prostaglandin E2.

Electroretinographic abnormalities in a rat glaucoma model with chronic elevated intraocular pressure.

The purpose of this study was to determine the ability of electroretinographic (ERG) measurements to document progression of the retinopathy in a rat glaucoma model. Thirty four rats with a chronic intraocular pressure (IOP) elevation induced in one eye by cautery of three episcleral/extra-orbital veins were studied in four separate groups. ERGs were recorded sequentially in Group A rats (n = 12) at baseline, and after approximately 20, 40 and 60 days of high IOP, and in three additional groups of rats (n = 6 or 10 per group) after approximately 58, 30 and 175 days of high IOP, respectively. Scotopic ERG parameters recorded simultaneously from both eyes in Group A rats were: a- and b-wave amplitudes, implicit times, oscillatory potential amplitudes (OPs) determined at three different light-flash intensities, and the light-adapted (photopic) ERG b-wave amplitude. In the other groups of rats, only scotopic ERG a-wave, b-wave and OP amplitudes were measured.In Group A rats that were followed sequentially, all the ERG parameters recorded with attenuated stimuli showed significant time-dependent changes in glaucomatous eyes relative to their contralateral normal eyes, with OPs showing the earliest significant difference after only 3 weeks of high IOP. When different groups of unilateral glaucomatous rats were compared beyond 8 weeks of elevated IOP only the OPs showed a continued decrease with time and good discrimination between glaucoma and normal eyes. Over a 25 week period of high IOP the scotopic OPs measured with attenuated light stimuli declined at the rate of approximately 1.5% per week and provided the best ERG measure to monitor progression of retinal pathophysiology in the vein-occlusion rat glaucoma model.

Maintaining mitochondrial membrane impermeability. an opportunity for new therapy in glaucoma?

Apoptosis may contribute to retinal ganglion cell loss in glaucoma and glaucoma models. Recent research has suggested that mitochondrially dependent apoptosis signaling may contribute to apoptosis in a rat model of glaucoma involving chronic increases in intraocular pressure. In some forms of apoptosis, mitochondrially dependent signaling involves increases in mitochondrial membrane permeability and the mitochondrial release of factors that signal for cell degradation. Opening of a multi-protein, mitochondrial megapore is one factor that contributes to the increased permeability and some anti-apoptotic proteins, particularly BCL-2 and BCL-X(L), bind at the megapore and facilitate megapore closure and reduce increases in mitochondrial membrane permeability. Phosphorylated protein kinase B (Akt) serves as an integrator for cellular survival signals and facilitates the megapore actions of BCL-2 and BCL-X(L), which could protect retinal ganglion cells against insults that induce apoptosis. Several anti-apoptotic agents are being evaluated for use in glaucoma, including brimonidine and propargylamines, which oppose mitochondrially dependent apoptosis through pathways involving phosphorylated Akt.

Retinal damage after 3 to 4 months of elevated intraocular pressure in a rat glaucoma model.

PURPOSE: To characterize a long-term elevated intraocular pressure (IOP) glaucoma model in the rat with respect to electroretinographic (ERG) changes and the pattern and mechanism of retinal ganglion cell (RGC) death. METHODS; An approximate doubling of IOP was induced in one eye (G) of female Wistar rats (150-180 g) by cautery of 3 episcleral/limbal veins. At intervals over 3 to 4 months, measurements of IOP and ERG changes (contact-lens electrode) were made in both the G and contralateral normal (N) eyes. At the end of 3 to 4 months of elevated IOP, RGCs were fluorescently labeled with Fluorogold (retrogradely from the superior colliculus), or retinas were labeled by intravitreal injection of a mitochondrial potential indicator dye and stained for apoptotic nuclei with a DNA dye. Flatmounts of fixed, dye-labeled retinas were examined by epifluorescence, confocal, or interference contrast microscopy. RESULTS: Elevated IOP was consistently maintained for up to 4 months in G eyes, but ERG a- and b-waves showed a statistically significant decline, of 30% to 40% in amplitude, after 3 months. Loss of RGCs in G retinas was primarily focal with no statistically significant loss demonstrable outside of the focal areas when assessed by an area sampling method for counting RGCs, which totaled 2% to 3% of the entire retinal area. Mitochondrial membrane potential of cells in the RGC layer was reduced by 17.5% (P: < 0.05) in regions surrounding areas of focal loss compared with comparable locations in control N eyes. After 3.5 months' elevated IOP the G retinas showed cell nuclei at various stages of apoptosis, from initial DNA condensation to fragmentation. CONCLUSIONS: The three-vein episcleral/limbal vein occlusion model for inducing glaucomatous pathology in the rat eye gives a consistent long-term elevation of IOP. After 3 to 4 months of approximately 100% increased IOP, the ERG responses begin to decline, there is a variable focal loss of RGCs, and some of the remaining RGCs show characteristics of stress and apoptosis. These changes seem consistent with retinal damage in human glaucoma (focal field defects), and this rat model appears to mimic some features of primary open-angle glaucoma.

The effect of divalent cations on bovine retinal NOS activity.

The divalent cation requirements of NOS activity in bovine retina homogenate supernatant were investigated. Supernatants were assayed under standard conditions (in mM: EDTA 0.45, Ca2+ 0.25, Mg2+ 4.0). In order to investigate the enzyme's dependence on divalent cations, the tissue homogenate was depleted of di- and trivalent cations by passing it over a cation-exchange column (Chelex 100). Surprisingly, NOS activity was 50-100% higher in this preparation. However, addition of either EDTA (33 microM) or EGTA (1 mM) almost fully inhibited NOS activity, suggesting a requirement for residual divalent metal cation(s). Phenanthroline or iminodiacetic acid at low concentrations had little effect on activity, suggesting no requirement for Fe2+, Zn2+ or Cu2+. Ca2+ had a moderate stimulatory effect, with an optimum activity around 0.01 mM. Mg2+ or Mn2+ had little effect at concentrations < 0.25 mM. However, in the presence of EDTA, Mn2+ or Ca2+ markedly stimulated NOS activity with the optimum at 0.1 mM. At high concentrations (> 0.1-0.2 mM), all divalent cations tested (Ba2+, Zn2+, Co2+, Mn2+, Mg2+, Ca2+), as well as La3+, dose-dependently inhibited NOS activity. We propose that retinal NOS requires low concentrations of naturally occurring divalent metal ions, most probably Ca2+, for optimal activity and is inhibited by high di- and trivalent metal concentrations, probably by competition with Ca2+.

Effect of latanoprost or 8-iso prostaglandin E2 alone and in combination on intraocular pressure in glaucomatous monkey eyes.

OBJECTIVE: To evaluate the possible additivity of the effects of latanoprost and 8-iso prostaglandin E2 (8-iso PGE2) on intraocular pressure (IOP) in monkey eyes with laser-induced glaucoma. METHODS: The IOP was measured hourly for 6 hours beginning at 9:30 AM on day 1 (baseline day), days 6 and 7 (single-agent therapy), and days 13 and 14 (combination therapy with both agents). Following 1 day of baseline measurement, 4 monkeys with unilateral glaucoma received monotherapy) twice daily with either 1 drop of 0.005% latanoprost, or 0.1% 8-iso PGE2, 25 microL, at 9:30 AM and 3:30 PM from days 2 through 7. From days 8 through 14, both agents were applied twice daily 5 minutes apart. RESULTS: The maximum reduction of IOP (mean +/- SEM) was 8.8 +/- 1.9 mmHg (26%) (P<.05) with latanoprost alone and 6.5 +/- 1.0 mmHg (21%) (P<.0l) with 8-iso PGE2 alone, 2 hours after the morning dosing on day 7. A further reduction of IOP of 4.0 +/- 0.6 mm Hg was produced when 8-iso PGE2 was added to latanoprost and of 3.0 +/- 0.7 mm Hg was produced when latanoprost was added to 8-iso PGE2 on day 13 before the morning dosing. Combination therapy with both agents caused maximum IOP reductions from baseline of 11.3 +/- 3.0 mm Hg (33%) (P<.05) (latanoprost with 8-iso PGE2 added) and of 9.8 +/- 1.3 mm Hg (31%) (P<.01) (8-iso PGE2 with latanoprost added) on day 14. CONCLUSION: Latanoprost and 8-iso PGE2 have an additive effect on IOP in glaucomatous monkey eyes. CLINICAL RELEVANCE: At least 50% of patients are treated with more than 1 ocular hypotensive medication. Thus, the determination of the additive effects on IOP of glaucoma medications will help to define optimum treatment regimens.

Light-induced retinal damage in mice carrying a mutated SOD I gene.

Transgenic mice expressing mutated mouse Cu/Zn superoxide dismutase (SOD I), corresponding to a mutation associated with familial amyotrophic lateral sclerosis, develop a fatal motorneuron degeneration that resembles the human disease. The biochemical properties of some mutant SOD I enzymes indicate that a gain of catalytic functions, (such as increased peroxidase activity) may be the pathologic factor(s). However, at the present time there is little in vivo evidence that a mutation-induced change in the catalytic activity of SOD I is directly involved in neuronal cell death or that vulnerability to cell death is related to the level of functional/metabolic activity of cells carrying mutated SOD I. In pigmented mice carrying the G86R mutation of mouse SOD I, exposure to constant bright light for 20 days caused a diminution of electroretinographic activity and specific degeneration of photoreceptor cells, while no pathological effects were seen in transgenic littermates not exposed to bright light or in light exposed non-transgenic littermates. These findings are the first to indicate that one mechanism for neuronal cell death by mutated SOD I is use-dependent and/or related to metabolic activity, and therefore may be due to a gain in function of catalytic activities involving superoxide/hydrogen peroxide. The light-exposure pathology in this transgenic mouse model indicates an essential role for SOD I in the protection of photoreceptors from light-damage.

[Ocular pulse amplitude during manipulation of systemic perfusion parameters]. / Okuläre Pulsamplitude bei gezielter Beeinflussung systemischer Perfusionsparameter.

BACKGROUND: The mechanism for the control of choroidal perfusion is unclear. The present study was designed to investigate for the effect of standardized alteration of systemic perfusion parameters on choroidal hemodynamics in healthy volunteers. METHODS: Intraocular pressure (IOP; German abbreviation: IOD), Ocular Pulse Amplitude (OPA), systemic blood pressure, and pulse rate were measured, and ocular perfusion pressure was calculated in 14 otherwise non-smoking, healthy volunteers prior to and following exercise and smoking. RESULTS: Exercise significantly (sig., p < 0.05) increased pulse rate, systolic blood pressure and ocular perfusion pressure, while it sig. (p < 0.05) reduced IOP and diastolic blood pressure. However OPA was not sig. (p > 0.1) affected by changes in these parameters. Smoking sig. (p < 0.05) increased systolic blood pressure, diastolic blood pressure, pulse rate, and ocular perfusion pressure but did not sig. (p > 0.09) alter OPA. CONCLUSION: Despite affecting ocular and systemic perfusion parameters, exercise and smoking did not alter OPA, suggesting functional isolation i.e. autoregulation of the choroidal and/or ophthalmic artery circulation.

Effect of 8-iso prostaglandin E2 on aqueous humor dynamics in monkeys.

OBJECTIVE: To evaluate the effects of 8-iso prostaglandin E2 (8-iso PGE2; prosta-5,13-dien-1-oic acid,11,15-dihydroxy-9-oxo-,[5Z,8beta-11X,13E,15 S]-) on the intraocular pressure (IOP), outflow facility, and aqueous humor flow rates in normal monkeys and monkeys with glaucoma. METHODS: The IOP was measured before and as long as 6 hours after the topical application of 8-iso PGE2 to 1 eye of 6 normal monkeys and to the glaucomatous eye of 8 monkeys with unilateral laser-induced glaucoma. The pupil diameter was measured at the same times as the IOP measurements in the normal monkeys. Tonographic outflow facility and fluorophotometric flow rates of aqueous humor were measured in 6 normal monkeys before and after drug treatment. RESULTS: In normal monkeys, a single dose of 0.1% 8-iso PGE2 reduced (P<.01) the IOP for 4 hours in the treated eyes with a maximum (mean +/- SEM) reduction of 3.2 +/- 0.2 mm Hg, compared with the contralateral control eyes. The pupil size was smaller (P<.01) in the treated eyes by as much as 1.0 +/- 0.2 mm for 4 hours. In 8 glaucomatous monkey eyes, the application of 0.05% and 0.1% 8-iso PGE2 reduced the IOP (P<.01) for as long as 2 and 5 hours, respectively. The maximum reduction in the IOP was 4.6 +/- 0.8 mm Hg (0.05%) and 6.0 +/- 0.8 mm Hg (0.1%) compared with baseline measurements. The magnitude and duration of the ocular hypotensive effect were enhanced with twice-a-day administration for 5 consecutive days. Outflow facility in normal monkey eyes was increased (P<.05) by 48% in the treated eyes, and aqueous humor flow was unchanged (P>.10), compared with vehicle-treated contralateral control eyes. Mild eyelid edema, conjunctival edema, hyperemia, and discharge appeared in some eyes treated with the 0.1% drug concentration. CONCLUSIONS: The use of 8-iso PGE2 reduces the IOP in both normal and glaucomatous monkey eyes. An increase in outflow facility appears to account for most of the IOP reduction in normal monkeys. CLINICAL RELEVANCE: The application of 8-iso PGE2 may have potential for the treatment of glaucoma as an outflow facility-increasing drug.

[Ocular pulse amplitude in ocular hypertension and open-angle glaucoma]. / Okuläre Pulsamplitude bei okulärer Hypertension und verschiedenen Glaukomformen.

The present study was designed to investigate choroidal perfusion in primary open-angle glaucoma patients with statistically elevated (HPG) and statistically normal (NPG) intraocular pressure (IOP) and in ocular hypertensive volunteers (OHT) before and after application of topical antiglaucomatous drugs. When compared to age-matched healthy controls, ocular pulse amplitude (OPA) and IOP were significantly increased in OHT and unchanged in HPG; in NPG, the IOP was unchanged, and the OPA was significantly reduced. The topical antiglaucomatous drugs used in this study reduced IOP, but did not increase OPA. When compared to HPG and NPG, OHT subjects showed increased choroidal perfusion which may help to prevent glaucomatous optic nerve damage.

[Peak pulse blood volume in controlled modification of local perfusion parameters]. / Pulsgipfelblutvolumen bei gezielter Beeinflussung lokaler Perfusionsparameter.

BACKGROUND: Choroidal autoregulation is discussed controversially. The present study was designed to investigate for the effect of standardized manipulation of local perfusion parameters on choroidal hemodynamics in cynomolgus monkey eyes with lasersurgically induced glaucoma (CMG; German abbreviation: RA-LHDG). METHODS: CMG unilaterally received 2-3 laser treatments so as to develop the lasered-eye glaucoma model. Intraocular pressure (IOP; German abbreviation: IOD) and Ocular Pulse Amplitude (OPA) were measured and Peak Pulse Blood Volume (PPBV; German abbreviation: PGBV) was determined. RESULTS: In CMG PPBV was not significantly (sig., p > 0.05) altered in laser-treated eyes--despite a sig. (p < 0.05) increase in IOP. Untreated contralateral control eyes of monkeys with an IOP-increase > 5 mm Hg in laser-treated eyes showed a sig. (p < 0.05) increase in PPBV. CONCLUSION: Unchanged PPBV in laser-treated eyes of CMG and an increase in PPBV in control eyes point at a bilateral autoregulatory compensatory mechanism, which is more manifest in eyes with IOPs in the normal range.

[Peak pulse blood volume and topical antiglaucoma drugs in rhesus monkeys with experimental open angle glaucoma]. / Pulsgipfelblutvolumen und topische Antiglaukomatosa bei Rhesusaffen mit experimentellem Offenwinkelglaukom.

BACKGROUND: The present study was designed to investigate the effect of topical antiglaucoma drugs on Peak Pulse Blood Volume (PPBV; German abbreviation: PGBV) in cynomolgus monkey eyes without (CMNG; German abbreviation: RA-KTL) and with lasersurgically induced glaucoma (CMG; German abbreviation: RA-LHDG). METHODS: CMG unilaterally received 2-3 laser treatments so as to develop the lasered-eye glaucoma model. Intraocular pressure (IOP; German abbreviation: IOD) and Ocular Pulse Amplitude were measured and PPBV was determined until the glaucoma model had stabilized. Consecutively topical antiglaucoma drugs (epinephrine, paraaminoclonidine, pilocarpine, timolol) were investigated in 4-8 animals in CMNG and CMG eyes. RESULTS: IOP and PPBV were not significantly altered in CMNG. In the CMG eyes epinephrine and paraaminoclonidine did not significantly alter IOP or PPBV, whereas pilocarpine and timolol sig. (p < 0.01) reduced IOP and significantly (p < 0.05) increased PPBV. CONCLUSION: With respect to improved PPBV in the CMG eye the rank order of drug effectiveness is timolol > pilocarpine > paraaminoclonidine > epinephrine.

Effect of 5-methylurapidil, an alpha 1a-adrenergic antagonist and 5-hydroxytryptamine1a agonist, on aqueous humor dynamics in monkeys and rabbits.

PURPOSE: To evaluate the effects of 5-methylurapidil (5-MU) on intraocular pressure (IOP) and aqueous humor dynamics in female cynomolgus monkeys and albino rabbits. METHODS: IOP was measured by pneumatonometer prior to and up to 6 hours after AM administration of 5-MU to one eye of each of 8 normal monkeys and to the laser-induced glaucomatous eye of 8 monkeys. During single-dose and 5-day multiple-dose testing, pupillary diameter (PD) was measured at the same time and same intervals as IOP measurements in the normal monkeys. Outflow facility and aqueous humor flow rates were measured in 8 normal monkeys before and after treatment. Uveoscleral outflow was measured in 8 rabbits before and after treatment. RESULTS: In normal monkeys, unilateral topical application of 2 x 25 microliters of 1% or 2% 5-MU significantly (p < 0.05) reduced pupil size and IOP bilaterally as compared to baseline measurements. The reduction in IOP (mean +/- SEM, mmHg) was up to 2.8 +/- 0.7 (1% 5-MU) and 4.4 +/- 0.5 (2% 5-MU) in the treated eyes, and 2.3 +/- 0.8 (1%) and 3.0 +/- 0.7 (2%) in the contralateral eyes. In glaucomatous monkeys, the maximum reduction in IOP was 6.5 +/- 1.0 mmHg (1%) and 7.5 +/- 0.8 mmHg (2%). The ocular hypotensive effect increased over time with twice-daily administration for 5 days. Compared with baseline values, outflow facility and aqueous flow rates in the treated eyes of normal monkeys were increased (p < 0.01) by 51% and by 11%, respectively. Uveoscleral outflow was unaltered (p > 0.3) in rabbits compared with baseline values. Mild corneal edema, corneal punctate erosions, and conjunctival discharge occurred in some eyes treated with either 1% or 2% 5-MU. CONCLUSIONS: 5-Methylurapidil, an antagonist at the alpha 1A-adrenergic receptor subtype and an agonist at the 5-HT1A receptor subtype, lowers IOP predominantly by increasing outflow facility and may have potential for the therapy of glaucoma.

[Effect of nifedipine on ocular pulse amplitude in normal pressure glaucoma]. / Einfluss des Nifedipins auf die okuläre Pulsamplitude bei Normaldruckglaukom.

BACKGROUND: Ocular pulse amplitude (OPA) is reduced in normal tension glaucoma (NTG) patients when compared to non-glaucomatous, healthy control subjects. This might be related to a vasospastic reaction. The objective of this study was to determine if low OPA in NTG is associated with a vasospastic reaction and its response to vasodilation. METHODS: Nifedipine, a calcium channel blocker, vasodilator and systemic antihypertensive agent improves visual fields in NTG patients following acute and chronic dosing. The effect of 60 mg of daily orally administered nifedipine on OPA, intraocular pressure (IOP, German abbreviation: IOD), blood pressure (BP, German abbreviation: RR) and pulse rate (PR, German abbreviation: HF) were measured prior to and for 3 months after initiating nifedipine therapy in 32 NTG patients with and without a vasospastic reaction as manifested by a local cold exposure test. Before treatment, all patients had reduced OPA evaluated with the Langham Ocular Blood Flow System. RESULTS: During nifedipine treatment NTG patients with a vasospastic reaction showed a significant (p < 0.001) increase in OPA, whereas NTG patients without a vasospastic reaction showed no sig. (p > 0.05) change in OPA. CONCLUSION: There may be two different subgroups of NTG patients, those who have a vasospastic reaction and react to nifedipine, while others lack the ability to react to nifedipine or might have a different, non-vasospastic pathology. Calcium channel blockers and other vasodilators may be useful in the treatment of vasospastic NTG patients.

Reduced ocular pulse amplitude in low tension glaucoma is independent of vasospasm.

PURPOSE: A vascular basis for the pathogenesis of primary open angle glaucoma has been postulated for many years. Defects in the regulation of ocular blood flow may be the initiating factor in this group of multifactorial diseases. This study was designed to evaluate the effect of vasospasm on ocular pulse amplitude (OPA) in low tension glaucoma (LTG) patients. METHODS: OPA, using the Langham Ocular Blood Flow (OBF) System, applanation intraocular pressure (IOP), systemic blood pressure and heart rate were measured and vasospasm was determined by a fingernail capillary blood flow test. RESULTS: OPA (mmHg) in the LTG patients with a vasospastic reaction (LTG-V, 1.4 +/- 0.1, n = 17) was not significantly (p > 0.09) different when compared with non-vasospastic LTG patients (LTG-NV, 1.5 +/- 0.2, n = 15) but was significantly (p < 0.001) reduced in LTG-V and LTG-NV patients when compared with matched healthy control subgroups (2.3 +/- 0.2 and 2.4 +/- 0.3, respectively). IOP (mmHg) in LTG-V (13.3 +/- 0.4) and LTG-NV (13.2 +/- 0.5) patient groups was not significantly (p > 0.05) different when compared with each other, but was significantly (p < 0.05) lower when compared with matched control subgroups (15.0 +/- 0.3 and 15.2 +/- 0.4, respectively). Haemodynamic parameters were not significantly different from controls. CONCLUSION: The abnormality in choroidal perfusion indicated by a reduction in OPA in all LTG patients is unrelated to the presence or absence of vasospasm.

Influence of physical exercise and nifedipine on ocular pulse amplitude.

BACKGROUND: Ocular pulse amplitude (OPA) was measured to investigate the influence of peripheral vasoconstriction and vasodilation on choroidal perfusion in healthy volunteers and to determine whether low OPA in low-tension glaucoma (LTG) patients is associated with a vasospastic reaction and its response to the calcium channel blocker nifedipine. METHODS: OPA was determined using the Langham ocular blood flow (OBF) system, applanation intraocular pressure (IOP), systemic blood pressure, and heart rate were measured, and ocular perfusion pressure was calculated before and after exercise and smoking in 12 otherwise nonsmoking, healthy volunteers and prior to and for 3 months after initiation of nifedipine therapy in 32 LTG patients with and without a vasospastic reaction as manifested by a nailfold capillary blood flow test. RESULTS: Exercise significantly (P < 0.05) increased heart rate, systolic blood pressure and ocular perfusion pressure, while it significantly (P < 0.05) reduced IOP and diastolic blood pressure. However, OPA was not significantly (P > 0.1) affected by changes in these parameters. Smoking significantly (P < 0.05) increased systolic and diastolic blood pressure, heart rate, and ocular perfusion pressure but did not significantly (P > 0.09) alter OPA. There were two distinct LTG subtypes, with and without a vasospastic reaction. Only those with a vasospastic reaction showed a significant (P < 0.001) increase in OPA after 3 months of nifedipine treatment, while all other parameters tested were not significantly altered. CONCLUSION: Despite affecting ocular and systemic perfusion parameters, exercise and smoking did not alter OPA, suggesting functional isolation, i.e. autoregulation of the choroidal and/or ophthalmic artery circulation in healthy volunteers. Low OPA in LTG was increased by nifedipine only in vasospastic LTG patients, suggesting different, vasotonus-related pathologies. Calcium channel blockers and other vasodilators may be useful in vasoreactive LTG patients with reduced OPA.

Role of nitric oxide synthase isozymes in endotoxin-induced uveitis.

PURPOSE: The authors previously reported that in vitro treatment with N(G)-nitro L-arginine (L-NNA), an inhibitor of nitric oxide synthase (NOS), reduces aqueous humor (AH) protein and cellular infiltration in endotoxin-induced uveitis in the rat eye. The objective of the current study was to determine the role(s) of respective major forms (constitutive and inducible) of NOS by comparing the effects of relatively selective inhibitors of these NOS isozymes. METHODS: N(G)-nitro L-arginine (L-NNA), a relatively selective inhibitor for constitutive NOS (c-NOS), and N-iminoethyl L-ornithine (L-NIO), a more selective inhibitor for inducible NOS (i-NOS), were administered in vivo. Male Lewis rats were footpad injected with bacterial lipopolysaccharide (LPS, 200 microgram) and were injected intraperitoneally at 0 hours, 6 hours, or both, after LPS injection with 10 mg of NIO, NNA, or saline as a control. Nitric oxide synthase activity in the ocular tissue and AH protein and cell content were determined at various times after treatment with LPS. RESULTS: After in vivo treatment, L-NIO was found to be a more potent inhibitor than L-NNA for ocular i-NOS (87% versus 43% inhibition), and L-NNA was more potent than L-NIO for ocular c-NOS (81% versus 39%). Two injections of L-NNA, one at time 0 and one 6 hours after LPS injection, inhibited the AH protein increase by 71%, but L-NIO did so by only 30%. L-NNA inhibited cellular infiltration by 86%, whereas L-NIO had no significant effect on cellular infiltration. A significant inhibition of cellular infiltration and AH protein increase also was observed with a single injection of 10 mg of L-NNA but not of L-NIO when the inhibitors were given simultaneously with LPS. Thus, reduction of uveitis symptoms correlates with the inhibition of c-NOS. CONCLUSIONS: The constitutive form of NOS in ocular tissue, presumably in vascular endothelial cells, appears to play a critical role at the onset of the development of endotoxin-induced uveitis.

Isoproterenol inhibits rod outer segment phagocytosis by both cAMP-dependent and independent pathways.

PURPOSE: The authors studied the involvement of cAMP-dependent second messenger systems in the inhibition of rod outer segment (ROS) phagocytosis by isoproterenol (ISO) and forskolin (FSK) using two membrane-permeant analogs of cyclic adenosine monophosphate (cAMP), the Rp and Sp diastereoisomers of cyclic adenosine 3',5' monophosphothioate (cAMPS). Rp-cAMPS is a potent competitive inhibitor of cAMP-dependent protein kinase I and II (PKA I and II), whereas Sp-cAMPS is a potent activator of these enzymes. METHODS: ROS phagocytosis was quantitated in cultured rat RPE cells using a previously described double immunofluorescence assay. RESULTS: Sp-cAMPS showed a dose-dependent inhibition of ROS phagocytosis, whereas 100 microM Rp-cAMPS had no effect on this process. Rp-cAMPS fully prevented the inhibitory effect of Sp-cAMPS and FSK but was able to prevent only partially the inhibition of ROS phagocytosis induced by ISO. Isoproterenol plus FSK showed an additive effect on the inhibition of phagocytosis, suggesting that they act at two independent sites. However, ISO plus Sp-cAMPS or FSK plus Sp-cAMPS showed no additivity. CONCLUSIONS: Results suggest that FSK inhibits ROS phagocytosis by RPE cells through a cAMP-dependent pathway, whereas ISO inhibits ROS phagocytosis by RPE cells through cAMP-dependent and cAMP-independent pathways.

Studies of the ocular pulse in primates.

Ocular pulse amplitude (OPA) and intraocular pressure (IOP) were measured in groups of human subjects with primary open-angle glaucoma (POAG), ocular hypertension (OHT), low-tension glaucoma (LTG), retinal degenerations (RD), in OHT volunteers treated with single doses of 2% epinephrine, 4% pilocarpine, 0.5% timolol or 1% p-amino-clonidine, and in normal subjects before and after exercise. Compared to normal controls, OHT subjects showed significantly higher IOP and OPA, while OPA was significantly lower in ocular normotensive LTG and RD subjects groups. All drug treatments lowered IOP in OHT subjects, but did not change OPA significantly. Exercise in normal volunteers increased calculated ocular perfusion pressure by 22.5%, lowered IOP by 32%, but showed no significant change in OPA. When IOP was elevated > or = 5 mmHg in lasered monkey eyes peak pulse volume (PPV) was increased significantly in the unlasered eyes. Epinephrine 2% or 1% p-aminoclonidine lowered IOP moderately with no change in PPV, while treatment with 4% pilocarpine or 0.5% timolol reduced IOP and increased PPV significantly. The findings suggest that LTG may be associated with an ocular vascular abnormality. OPA in OHT or normal human subjects did not change when IOP was decreased by antiglaucoma drug treatments or exercise, respectively. These results indicate that OPA may be physiologically autoregulated in human subjects with IOPs in the 11-21 mmHg range. However, laser-induced glaucomatous monkey eyes with higher IOP (30-35 mmHg) did not autoregulate, but showed a low peak pulse volume, which increased when IOP was reduced 5 mmHg or more by means of antiglaucoma drug treatment.