Vehicle-treated Eyes Research Articles

Effect of Topical Dorzolamide on Tissue Circulation in the Rabbit Optic Nerve Head

Purpose: To examine the effect of 1% topical dorzolamide on tissue circulation in the optic nerve head (ONH) of Dutch rabbits. Methods: A laser speckle tissue circulation analyzer was used. One eye of each rabbit received 1% topical dorzolamide twice daily for 20 days, and the fellow eye received the vehicle in a masked, randomized manner. Intraocular pressure (IOP) was measured every 5 days. The normalized blur (NB) value, a quantitative index of tissue blood flow velocity in the ONH, was measured before treatment and 2 hours after the last instillation on the 20th day. Results: The IOP was lowered by about 2 mm Hg only in the dorzolamide-treated eyes ( P < .01). The NB value showed no significant change in either dorzolamide-treated or vehicle-treated eyes. Conclusions: Long-term topical dorzolamide does not affect the ONH tissue circulation in dorzolamide- and vehicle-treated eyes of Dutch rabbits.

Topical prostaglandin F2alpha treatment reduces collagen types I, III, and IV in the monkey uveoscleral outflow pathway.

Topical prostaglandin F2alpha isopropyl ester increases uveoscleral outflow in monkeys and humans. To investigate the effects of prostaglandin F2alpha isopropyl ester with topical administration on collagen types I, III, and IV within the anterior segment tissue of monkey eyes. Eight eyes of 4 cynomolgus monkeys were evaluated. One eye of each monkey was treated with 2 microg of prostaglandin F2alpha isopropyl ester twice daily for 5 days, and intraocular pressure reduction was confirmed. These eyes were fixed in methacarn, and paraffin sections were immunostained using antibodies to collagen types I, II, or IV. To measure staining intensity, optical density (OD) was determined using 2-dimensional imaging densitometry. Mean OD scores along line segments placed over the ciliary muscle were determined. Mean+/-SD OD scores for collagen types I, III, and IV were less in the ciliary muscle of prostaglandin-treated eyes than in vehicle-treated eyes by 52%+/-7%, 45%+/-6%, and 45%+/-5%, respectively. In the sclera adjacent to the ciliary body, mean OD scores for collagen types I and III were less in prostaglandin-treated eyes, by 43%+/-32% and 45%+/-13%, respectively. The scleral stroma was minimally immunoreactive for collagen type IV. All differences were significant by the paired Student t test (P<.05). This study shows reduced collagen types I, III, and IV immunoreactivity in the ciliary muscle and adjacent sclera following topical prostaglandin F2alpha isopropyl ester treatment. These reductions may contribute to the increased uveoscleral outflow observed with topical prostaglandin treatment. The cellular mechanism by which certain prostaglandins lower intraocular pressure is not known. The present study provides immunohistochemical data demonstrating that intraocular pressure reduction that occurs with topical prostaglandin F2alpha is associated with a reduction of collagens within the uveoscleral outflow pathway.

Clarithromycin for experimental Staphylococcus aureus keratitis

Purpose. Clarithromycin, a macrolide antibiotic not previously tested against the common causes of bacterial keratitis, was analyzed for its effectiveness in reducing the number of viable bacteria in a Staphylococcus keratitis model. An in vivo comparison of the effectiveness of clarithromycin to erythromycin, minocycline, and tetracycline for three strains of Staphylococcus aureus was done. Methods. Rabbit eyes were intrastromally injected with 100 colony forming units of one of three strains of S. aureus. Two strains were methicillin-sensitive (ATCC 25923 and MSSA 309) and one strain methicillin-resistant (COL). Eyes were treated every 30 minutes with 0.3% clarithromycin, erythromycin, tetracycline, or minocycline from 4 to 9 hours postinfection. The number of colony forming units (CFU) per cornea in all eyes was determined at 10 hours postinfection. Results. Vehicle-treated and untreated eyes (controls) contained over 6 logs of CFU per cornea, a value significantly higher than any of the antibiotic-treated eyes (P = 0.0001). Clarithromycin or erythromycin therapy significantly decreased the number of CFU per cornea by approximately 5 logs in the eyes infected with the methicillin-sensitive strains and by approximately 4 logs in the eyes infected with the methicillin-resistant strain. Tetracycline and minocycline were also successful in treating these strains, but overall showed less effectiveness than clarithromycin and erythromycin. Conclusions. Clarithromycin proved to be an effective ocular medication for the therapy of experimental S. aureus keratitis. The effectiveness of clarithromycin in this model and its known effectiveness for a variety of bacterial pathogens suggests a role for this drug as a useful ocular antibiotic.

Effect of Topical Carteolol on Iridial Circulation in Pigmented Rabbit Eyes

The effect of a single instillation of topical carteolol on iridial tissue circulation of pigmented rabbits was studied. The blood flow rate and a quantitative index of tissue blood velocity (NB iris) were measured simultaneously, using the microsphere technique and the laser speckle method, before and 2 hours after the instillation of 20 microliters of 2% carteolol or the vehicle. Consecutive changes of intraocular pressure and NB iris were also studied at 30-minute intervals for 2.5 hours after a single instillation of 2% carteolol in one eye and the vehicle in the contralateral eye. To provide a control, intraocular pressure and NB iris were measured according to the same schedule after the vehicle instillation in both eyes. Two hours after carteolol instillation, iridial blood flow rate and NB iris significantly increased to 127 ± 8% and 122 ± 9% (mean ± SEM, n = 8) of the baseline. Unilateral instillation of carteolol significantly reduced intraocular pressure by about 9 mm Hg in both the carteolol- and vehicle-treated eyes ( P < 0.001, analysis of variance); and NB iris was significantly increased by about 20% in both eyes ( P < 0.001, analysis of variance), compared with control eyes.

Lactoferrin suppresses loss of corneal epithelial integrity in a rabbit short-term dry eye model.

Human tear fluid contains lactoferrin at the highest concentration. In patients with dry eye such as Sjogren's syndrome, the concentration of lactoferrin in the tears is approximately half the normal value. The present study utilizes a short-term rabbit dry eye model to evaluate if lactoferrin containing eye drops can reverse any of the damage produced by blockage of blinking with an ocular speculum. Damage was evaluated based on the extent of methylene blue staining in histological sections. After 3 h of desiccation, the amount of extractable dye recovered following sacrifice increased by more than 4-fold in the vehicle-treated eyes. However, in those rabbits treated with 1% lactoferrin, dye recovery was only 40% of the value in the vehicle-treated eyes. Between 1-3 h and over a concentration range from 0.01 to 1% lactoferrin, the decreases in staining were both time and concentration dependent. Alternatively, if 1% lactoferrin was applied during the desiccation period, there was partial restoration of corneal epithelial integrity. These results suggest that lactoferrin may be of therapeutic value in decreasing the loss of corneal epithelial integrity in dry eye.

Experimental use of tetrodotoxin for corneal pain after excimer laser keratectomy.

To determine the duration of anesthesia, effect on corneal reepithelialization, and systemic toxicity of topical tetrodotoxin (TTX) administered after excimer laser keratectomy. Two groups of six rabbits each underwent excimer laser keratectomy in the right eye to create a 5-mm-diameter wound, 75 mm in depth. One group then received a 40-microl aliquot of topical 1 mM TTX into the injured eye, whereas the other group received 40 microl of the sodium citrate vehicle as a control. The rabbits were treated with TTX or vehicle again at 6, 12, 18, and 24 h. Corneal sensation was tested at 3, 6, 9, 12, 15, 18, 21, 24, 30, 32, and 40 h. To determine whether TTX inhibited corneal reepithelialization, compared with vehicle-treated control eyes, the healing rate of the epithelial defect was measured. Administration of TTX every 6 h for 24 h produced nearly complete anesthesia for > or = 30 h. At 32 h, 8 h after the final application of TTX, there was still significant anesthesia of the TTX-treated corneas (p = 0.0325, Wilcoxon test). Normal corneal sensation in all TTX-treated animals returned at 40 h, or 16 h after the final dose. In contrast, vehicle-treated eyes all had normal sensation for nearly the entire duration of the experiment. At 40 h, the TTX-treated eyes had slightly larger defects than vehicle-treated eyes, 7.85+/-1.74 versus 4.54+/-1.31 mm2 (p < 0.025, t test). However, at 49 h and thereafter, both groups were equally healed (p > 0.05, t test). No systemic toxicity was observed in any of the rabbits. Topical TTX is a long-acting and nontoxic local anesthetic in a rabbit model of excimer laser keratectomy.

Reduction by anti-inflammatory drugs of the response of corneal sensory nerve fibers to chemical irritation: Chen X, Gallar J, Belmonte C. Invest Ophthalmol Vis Sci 1997;38:1944–1953

Nonsteroidal antiinflammatory drugs (NSAIDs) have been applied topically to reduce ocular pain caused by corneal injury or anterior segment surgery. The authors investigated whether the analgesic effects of the NSAIDs diclofenac, indomethacin, and flurbiprofen and of the calcium channel antagonist diltiazem on corneal pain are mediated by a reduction of nerve activity in corneal polymodal nociceptive fibers.Impulse activity of single A-delta and C corneal nerve fibers was recorded from the ciliary nerves of anesthetized cats. Polymodal units were identified by their response to both touching with the Cochet-Bonnet esthesiometer and to acidic stimulation with 30-second pulses of 80% or 98.5% CO2 or 60 microl of 10 mM acetic acid, applied to the corneal receptive area. Ongoing impulse activity, firing responses to CO2 or acetic acid, and mechanical threshold of single fibers were recorded before and at various times (5 to 90 minutes) after topical application of 0.1% sodium diclofenac, 0.03% sodium flurbiprofen, 0.1% indomethacin, and 0.045% diltiazem hydrochloride or of their vehicles.Indomethacin, diclofenac, and flurbiprofen, in decreasing order of potency, gradually reduced the mean frequency of the impulse response of corneal polymodal nerve fibers evoked by CO2 stimuli. The progressive increase of ongoing activity, observed in vehicle-treated eyes after repeated CO2 stimulation was also prevented by NSAIDs. Diltiazem also attenuated the response to CO2 for a shorter period of time and with a faster time course. The mechanical threshold of corneal polymodal fibers was not affected by treatment with any of these drugs.Indomethacin, diclofenac, and flurbiprofen, as well as the calcium antagonist diltiazem, diminish the responsiveness of corneal polymodal nociceptors to chemical stimuli. This appears to be caused, in part, by a direct effect of these drugs on the excitability of polymodal nerve endings, but also by an inhibition by NSAIDs of the formation of cyclooxygenase products such as prostaglandins, thus reducing the enhanced responsiveness of nociceptors caused by local release of arachidonic acid metabolites from injured cells.

Effect of topical carteolol on tissue circulation in the optic nerve head

The effect of topical 2% carteolol on tissue circulation in the albino rabbit optic nerve head (ONH) was investigated using a laser speckle tissue circulation analyzer. In the first experiment, the normalized blur (NB) value, a quantitative index of tissue blood flow velocity in the ONH, intraocular pressure (IOP), blood pressure (BP), and pulse rate were measured under general anesthesia before as well as 30, 60, 90, and 120 minutes after a 20-μL instillation of carteolol in one eye and the vehicle in the other eye in a masked, randomized manner. In the second experiment, one eye of a rabbit received carteolol twice daily for 20 days and the fellow eye received the vehicle in a masked, randomized manner. The IOP was measured every 5 days, and the NB in the ONH and IOP were measured before treatment and 2 hours after the last instillation on the 20th day. After a single instillation of carteolol, pulse rate showed a maximum reduction of 15%, and IOP in the carteolol-treated eyes showed a maximum decrease of 22%. The NB in the ONH and BP did not show any significant change during the experiment. After 20-day treatment with carteolol, IOP showed a maximum decrease of 25% in the carteolol-treated eyes and 21% in the vehicle-treated eyes. The NB showed a significant increase of 15% ( P < 0.01) in the carteolol-treated eyes and 11% ( P < 0.01) in the vehicle-treated eyes. It was indicated that long-term topical carteolol increased the blood velocity in the ONH tissue both in the carteolol- and vehicle-treated contralateral eyes in albino rabbits.

Ocular pulse amplitude and local carbonic anhydrase inhibition

Ocular perfusion is increasingly being discussed in the pathogenesis of glaucoma. The present study was designed to investigate ocular pulse amplitude (OPA) in primary open-angle glaucoma (POAG) patients with elevated intraocular pressure (HTG) and non-glaucomatous controls (CTL) following topical application of the carbonic anhydrase inhibitor dorzolamide. OPA (Ocular Blood Flow System, OBF Labs UK) intraocular pressure (IOP), heart rate (HR), and systolic (BPsyst) and diastolic (BPdiast) bronchial artery pressures were measured before and 2 days after initiating treatment in 33 cataract patients with n = 14) and without (n = 19) POAG. Following application of dorzolamide, IOP (mmHg) in drug-treated HTG and CTL eyes was highly significantly reduced (p < 0.001) and in vehicle-treated HTG and CTL eyes significantly reduced (p < 0.03) compared to respective baseline measurements. OPA (mmHg) in drug-treated HTG and CTL eyes was significantly increased (p < 0.05) and in vehicle-treated eyes in affected compared to respective baseline measurements. Systemic perfusion parameters were also unchanged. Dorzolamide increased OPA in HTG and CTL eyes. An increase in OPA may improve the prognosis of HTG.

Effect of topical timolol on tissue circulation in optic nerve head

The effects of topical 0.5% timolol on tissue circulation in the albino rabbit optic nerve head (ONH) were investigated using a laser speckle tissue circulation analyzer. In the first experiment, the normalized blur (NB) value, a quantitative index of tissue blood flow velocity in the ONH, intraocular pressure (IOP), blood pressure, and pulse rate were measured under general anesthesia before, and 30, 60, 90, and 120 minutes after a 20 μL instillation of timolol in one eye and the vehicle in the other eye in a masked, randomized manner. In the second experiment, one eye of a rabbit received timolol twice daily for 20 days and the fellow eye received the vehicle in a masked, randomized manner. Every 5 days IOP was measured and the NB in the ONH and IOP were measured before treatment and 2 hours after the last instillation on the 20th day. After a single instillation of timolol, PR showed a maximum reduction of 12% and IOP in the timolol-treated eyes showed a maximum decrease in 25%. NB in the ONH and BP did not show any significant change during the experiment. After a 20-day treatment with timolol, IOP showed a maximum decrease of 25% in the timolol-treated eyes and 16% in the vehicle-treated eyes. The NB in the timolol-treated eyes increased significantly by 16% ( P < 0.01), whereas that in the vehicle-treated eyes showed no significant change. It was suggested that long-term topical timolol with a normal drug regimen caused a significant increase in the peripheral blood velocity in the ONH only in the timolol-treated eyes, at least partly, by local penetration of the drug. Ocular penetration of topically applied timolol is thought to be similar between rabbit and human eyes. Therefore, the present results may have clinical implications.

Effects of Topical Adrenergic Agents on Tissue Circulation in Rabbit and Human Optic Nerve Head Evaluated With Laser Speckle Tissue Circulation Analyzer

The effects of topical adrenergic agents on the tissue circulation of the optic nerve head (ONH) were studied in animal and human eyes with use of a laser speckle tissue circulation analyzer. Drugs studied were nipradilol, a β1–2-blocker with weak α-blocking and nitroglycerinlike activities; bunazosin, a selective α1-blocker; carteolol, β1–2-blocker with intrinsic sympathomimetic activity; and betaxolol, a selective β1-blocker. In the animal experiment, one eye each of nine albino rabbits received 0.25% nipradilol (N = 9) twice daily for 15 days or 0.01% bunazosin (N = 10) twice daily for 20 days; the fellow eye received the vehicle of each drug in a randomized masked design. Normalized blur (NB) in an area of ONH free of visible surface vessels, a quantitative index of peripheral blood velocity in ONH, was measured under general anesthesia before treatment and 2 hours after the last instillation on the 15th or 20th day. At 15 days, intraocular pressure and NB in the nipradilol-treated eyes decreased by 4.3 mm Hg (P < 0.01) and increased by 15.9% (P < 0.01), respectively, while those in the vehicletreated eyes showed little change. At 20 days NB in both bunazosin- and vehicle-treated eyes showed little change, while intraocular pressure decreased only in the bunazosin-treated eyes (P < 0.01). In the human volunteer experiment NB was averaged over five pulses (mean NB) in both eyes before and 1.5, 3.0 and 4.5 hours after a 30 μL instillation of the vehicle of carteolol or betaxolol to serve as a control. Intraocular pressure, blood pressure and pulse rate were also measured. One week later a 30 μL drop of 2.0% carteolol (N = 6) or 0.5% betaxolol (N = 10) was instilled in one randomly chosen eye and the vehicle for each drug in the other eye, and those parameters were measured as above in a doublemasked manner. In the carteolol experiment, mean NB in both eyes was significantly higher at 3 hours than it was in the control experiment (P < 0.05), while intraocular pressure was significantly lower in both eyes and blood pressure and pulse rate showed little change. In the betaxolol experiment, intraocular pressure at 1.5 hours and blood pressure at 4.5 hours were significantly lower than those in the control experiment (P < 0.05), while mean NB and pulse rate showed little change. In summary, using the laser speckle method we found that topical nipradilol and carteolol increase ONH blood velocity in rabbits and humans, respectively. This finding suggests that some topical P-blockkes used clinically may influence the ONH tissue circulation in patients and that this method may be used in clinical situations to assess the effect of various drugs on ONH circulation.

Effect of topical beta-blockers on tissue blood flow in the human optic nerve head

PURPOSE. To study the effect of topical 0.5% timolol and 2% carteolol on tissue blood flow in the human optic nerve head (ONH). METHODS. Using a laser speckle tissue blood flow analyzer, normalized blur (NB), a quantitative index of tissue blood velocity, was measured every 0.125 s in the temporal site of the ONH free of visible surface vessels and averaged over 3 cardiac pulses (NB ONH) . To serve as a baseline, NB ONH and intraocular pressure (IOP) in both eyes, blood pressure (BP) and pulse rate (PR) were recorded in healthy volunteers before, 1.5, 3 and 4.5 hrs after a 30L instillation of the vehicle of timolol or carteolol. From the following day and twice daily for 3 weeks, 30L of either 0.5% timolol or 2% carteolol was instilled into one eye and the respective vehicle into the fellow eye in a masked manner. NB ONH, IOP, BP and PR were again recorded on the 21st and last experiment day. IOP was also recorded on the 7th and 14th days. Carteolol concentration in the plasma was also recorded after instillation of carteolol on the 21st day. RESULTS. During the baseline experiments, all the parameters recorded showed no significant change. After topical timolol, IOP was significantly reduced bilaterally with more reduction in the timolol-treated eye. Bilateral NB ONH, BP and PR showed little change on the 21st day. After topical carteolol, IOP was significantly reduced bilaterally with more reduction in the carteolol-treated eyes on the 21st day. NB ONH in the carteolol- and vehicle-treated eyes was significantly higher on the 21st day than recorded in the same eye in the baseline experiment (P = 0.013 and 0.047), while BP and PR showed little change. The maximum carteolol concentration in plasma at 3 hrs on the 21st day averaged 1294 pg/ml. CONCLUSIONS. Results indicated that 3-week twice daily topical timolol treatment had no deleterious effect on the ONH tissue blood flow in the human eye, and that 3-week twice daily topical carteolol treatment may increase the tissue blood flow in the human ONH.

Treatment with bifemelane for optic nerve damage following high intraocular pressure in rat eyes.

This study found that pretreatment with 4-(o-benzylphenoxy)-N- methylbuty-lamine hydrochloride (bifemelane hydrochloride, Celeport) reduced ischemia-reperfusion injury in rat eyes. Bifemelane (25 mg/kg) was injected intraperitoneally 30 minutes before an ischemic insult, then acute ischemia of the retina and optic disc was induced by increasing intraocular pressure to 110 mmHg for 45 minutes. After one week, the axonal count of the optic nerve was investigated using electron microscopy. The control group consisted of vehicle-treated eyes which received normal saline. The axon count was 93.4 +/- 7.9 for the bifemelane treated group, and 79.2 +/- 6.4 for the controls. The axon count in the treated group was significantly higher. These results suggest that bifemelane, which prevents cerebral nerve cell damage from ischemia, can reduce ischemic retinal nerve cell injury.

Effects of endothelin A and B receptors on aqueous humor dynamics in the rabbit eye.

This study attempted to clarify the role of endothelin A and B (ETA and ETB) receptors in modulating aqueous humor dynamics in the rabbit eye. Intraocular pressure (IOP), aqueous flow, total outflow facility, and uveoscleral outflow were measured before, and 24 hours after, intravitreal injection (20 microliters) of a selective ETB agonist, sarafotoxin S6c (SRTX S6c, 10(-5) M), into one eye and its vehicle into the contralateral eye. The measurements were also performed before and after injection of a selective ETA antagonist, 97-139 (10(-2) M) + ET-1 (10(-5) M) into one eye, and vehicle + ET-1 (10(-5) M) into the contralateral eye. In the SRTX S6c-treated eye, total outflow facility increased significantly by 106% compared with that in the vehicle-treated eye 24 hours after injection; aqueous flow and uveoscleral outflow failed to change significantly. The ETA antagonist 97-139 inhibited the decrease in aqueous flow partially, but significantly, but failed to inhibit the increase in total outflow facility caused by ET-1. Therefore, both ETA and ETB play a significant role in modulating aqueous humor dynamics in the rabbit eye. ETA can modulate aqueous humor production, at least in part. ETB can modulate total outflow facility, and the possibility for ETB in modulating aqueous humor formation cannot be ruled out.

Effect of daily topical ethacrynic acid on aqueous humor dynamics in monkeys.

We determined the effect of 1.5% ethacrynic acid (ECA) in ointment on intraocular pressure (IOP) and outflow facility following 5 days of topical treatment in ocular normotensive cynomolgus monkeys. Twelve monkeys received a 1 cm strip of ointment containing ECA in one eye but without ECA in the other once daily for 5 days. On Day 1 and Day 5 IOP was measured immediately before and 1 and 3 h after treatment. Outflow facility (perfusion) was determined 3.5 h after treatment on Day 5. The ECA-vehicle IOP differences averaged -2.8 +/- 0.8 (S.E.M) (p < 0.01), -1.7 +/- 0.6 (p < 0.02) and -3.7 +/- 0.7 mm Hg (p < 0.001), equivalent to IOP reductions of 21 +/- 6% (p < 0.01), 13 +/- 5% (p < 0.02) and 26 +/- 5% (p < 0.002) respectively, at 0, 1 and 3 h respectively after treatment on Day 5. Facility averaged 40 +/- 15% higher (p < 0.03) in the ECA-treated compared to the vehicle-treated eyes 4 h after treatment on Day 5.

Effects of PhXA41, A New Prostaglandin F2α Analog, on Aqueous Humor Dynamics in Human Eyes

PhXA41, a new phenyl-substituted analog of a prostaglandin F2 alpha (PGF2 alpha) prodrug (13,14-dihydro-17-phenyl-18,19,20-trinor-prostaglandin F2 alpha-1-isopropyl ester), is an effective ocular hypotensive agent in patients with glaucoma. To understand its mechanism of action, various components of aqueous humor dynamics were examined after topical application to human eyes. In a randomized, double-masked, placebo-controlled study, PhXA41 (0.006%) was given topically twice daily for 1 week to one eye each of 22 volunteers with normotension or ocular hypertension. The other eye was similarly treated with vehicle. Intraocular pressure (IOP) was measured by pneumatonometry and tonographic outflow facility by pneumatonography. Aqueous flow and outflow facility were determined either directly or indirectly by a fluorophotometric technique, and uveoscleral outflow was calculated secondarily. Comparison of values obtained in treated versus contralateral control eyes and on baseline versus day 8 of treatment were made. Compared with baseline measurements, PhXA41 significantly (P < 0.001) reduced IOP by 5.5 +/- 0.6 mmHg (mean +/- standard error of the mean) as measured 3 hours after the last dose on the eighth day of treatment. Aqueous flow, tonographic outflow facility, and fluorophotometric outflow facility were not changed by PhXA41. However, uveoscleral outflow was significantly greater in the PhXA41-treated eyes (0.87 +/- 0.22 microliter/minute) compared with either the contralateral vehicle-treated eyes (0.14 +/- 0.30; P < 0.02) or baseline measurements (0.39 +/- 0.20 microliter/minute; P < 0.05). PhXA41 decreases IOP in humans by increasing uveoscleral outflow without significantly affecting other parameters of aqueous humor dynamics.

Clinical evaluation of ketorolac tromethamine 0.5% ophthalmic solution for the treatment of seasonal allergic conjunctivitis

We evaluated 148 patients with allergic conjunctivitis in a double-masked, paired comparison clinical trial comparing ketorolac 0.5% ophthalmic solution with vehicle. Patients received one drop of each study medication in preassigned eyes, four times a day, for seven days. Both treatments showed significant changes from baseline in the signs and symptoms associated with allergic conjunctivitis. Evaluations at the final visit (day 7 or 8) showed that ketorolac-treated eyes had a significant treatment response when compared to vehicle-treated eyes for conjunctival inflammation (p = 0.010), ocular itching (p = 0.006), swollen eyes (p = 0.002), discharge/tearing (p = 0.021), foreign body sensation (p = 0.035), and conjunctival injection (p = 0.016). Mean scores evaluating the overall therapeutic effect of the study treatments at the completion of the study were higher for ketorolac-treated eyes than for vehicle-treated eyes as rated by investigators (p = 0.004) and study patients (p < 0.001). Results of this study confirmed the trends of a previous study showing that ketorolac 0.5% ophthalmic solution applied topically is an effective therapy for allergic conjunctivitis.

Delay of ganglion cell death by tetrodotoxin during retinal development in chick embryos

Neuronal activity plays an important role in the formation of topographic projection in the nervous system. In order to examine the role of neuronal activity on the retinotectal projection system, tetrodotoxin (TTX) was injected into one eye ball of the chick embryo once between day 9 and 14 of incubation and the number of dying retinal ganglion cells (RGCs) stained with Nile blue in the whole-mounted retinae were counted. In the control retinae, dying RGCs first appeared at stage 36 and disappeared after stage 43. The peak of RGC death was at stage 38 in the temporal and the central parts and at stage 39 in the nasal part. The total amount of dying RGCs in the temporal part was less than that in the central and the nasal parts. After injection of TTX into one eye ball, the peak of RGC death was at stage 40, which was 2 days later than that in the control retinae. TTX treatment did not affect the total amount of dying RGCs in the treated eye, but resulted in their decrease in the vehicle-treated eye which was contralateral to TTX treatment. These findings indicate that inhibition of neuronal activity of RGCs by TTX treatment delays the timing of RGC death.

Perspectives on aqueous humor formation

The effects of topical adrenergic agents on the tissue circulation of the optic nerve head (ONH) were studied in animal and human eyes with use of a laser speckle tissue circulation analyzer. Drugs studied were nipradilol, a β1–2-blocker with weak α-blocking and nitroglycerinlike activities; bunazosin, a selective α1-blocker; carteolol, β1–2-blocker with intrinsic sympathomimetic activity; and betaxolol, a selective β1-blocker. In the animal experiment, one eye each of nine albino rabbits received 0.25% nipradilol (N = 9) twice daily for 15 days or 0.01% bunazosin (N = 10) twice daily for 20 days; the fellow eye received the vehicle of each drug in a randomized masked design. Normalized blur (NB) in an area of ONH free of visible surface vessels, a quantitative index of peripheral blood velocity in ONH, was measured under general anesthesia before treatment and 2 hours after the last instillation on the 15th or 20th day. At 15 days, intraocular pressure and NB in the nipradilol-treated eyes decreased by 4.3 mm Hg (P < 0.01) and increased by 15.9% (P < 0.01), respectively, while those in the vehicletreated eyes showed little change. At 20 days NB in both bunazosin- and vehicle-treated eyes showed little change, while intraocular pressure decreased only in the bunazosin-treated eyes (P < 0.01). In the human volunteer experiment NB was averaged over five pulses (mean NB) in both eyes before and 1.5, 3.0 and 4.5 hours after a 30 μL instillation of the vehicle of carteolol or betaxolol to serve as a control. Intraocular pressure, blood pressure and pulse rate were also measured. One week later a 30 μL drop of 2.0% carteolol (N = 6) or 0.5% betaxolol (N = 10) was instilled in one randomly chosen eye and the vehicle for each drug in the other eye, and those parameters were measured as above in a doublemasked manner. In the carteolol experiment, mean NB in both eyes was significantly higher at 3 hours than it was in the control experiment (P < 0.05), while intraocular pressure was significantly lower in both eyes and blood pressure and pulse rate showed little change. In the betaxolol experiment, intraocular pressure at 1.5 hours and blood pressure at 4.5 hours were significantly lower than those in the control experiment (P < 0.05), while mean NB and pulse rate showed little change. In summary, using the laser speckle method we found that topical nipradilol and carteolol increase ONH blood velocity in rabbits and humans, respectively. This finding suggests that some topical P-blockkes used clinically may influence the ONH tissue circulation in patients and that this method may be used in clinical situations to assess the effect of various drugs on ONH circulation.

Stimulation of Tear Secretion and Treatment of Dry-Eye Disease With 3-Isobutyl-1-methylxanthine

We examined the effect of topically applied 3-isobutyl-1-methylxanthine (IBMX), a known secretagogue, on tear secretion and dry-eye disease in a clinical study. We found that IBMX produced a dose-dependent decrease in tear film osmolarity that was significant at 3.0 mmol/L (P less than .0005) in patients with dry-eye disease. This effect was not blocked by prior administration of proparacaine hydrochloride (P less than .05). Throughout a 4-week, open-label, vehicle-controlled study, IBMX decreased tear film osmolarity significantly, whereas vehicle alone did not. After 4 weeks, mean (+/- SEM) osmolarity in IBMX-treated eyes decreased from 325 +/- 3.2 mOsm/L to 312 +/- 1.8 mOsm/L but remained unchanged in vehicle-treated eyes (323 +/- 4.4 mOsm/L vs 320 +/- 4.2 mOsm/L). In our study, IBMX was significantly more effective than vehicle alone in decreasing rose bengal staining (P less than .02). Hence, topical IBMX stimulated tear secretion and decreased ocular surface disease in patients with dry-eye disease.