Tear production after unilateral removal of the main lacrimal gland in squirrel monkeys.

OBJECTIVE: To study the effects of lacrimal gland removal on basal and reflex tear production and on the ocular surface in the squirrel monkey. METHODS: Unilateral main lacrimal gland removal in 6 squirrel monkeys was followed by Schirmer testing, slit-lamp examination with fluorescein, and collection of basal and reflex (stimulated) tears for analysis of tear protein spectra between 0 and 20 kd, as well as histological evaluation. RESULTS: Schirmer test results showed an 80% decrease in basal tears and a 90% decrease in reflex tears during week 1, and a 32.2% and 33.3% decrease, respectively, at week 20 after surgery, compared with the contralateral control side. However, no gross abnormalities or fluorescein staining were seen in 5 of the 6 monkeys, and the conjunctival surfaces remained normal. The main and accessory lacrimal glands appeared to secrete similar types of proteins. No histological changes were seen in corneal, conjunctival, or eyelid tissues 20 weeks after surgery. CONCLUSIONS: Tears from accessory lacrimal glands were sufficient to maintain a stable tear layer on the cornea, suggesting that so-called basal tear flow is made up of fluid from both main and accessory lacrimal glands and that decreased tear production by the main lacrimal gland is not a causative factor in keratoconjunctivitis sicca. CLINICAL RELEVANCE: This study shows that total removal of the main lacrimal gland does not in itself lead to keratoconjunctivitis sicca. However, the nature of neural control of the accessory glands is not yet clear.

[Comparative study of tears protein composition in patients after photorefractive keratectomy and laser specialized keratomileusis]. / Sravnitel'noe issledovanie belkovogo sostava slezy patsientov posle fotorefraktsionnoi keratoéktomii i lazernogo spetsializirovannogo keratomileza.

Changes in the protein spectrum of the lacrimal fluid, resultant from refraction ocular surgery (photorefraction keratectomy, PRK, and laser specialized keratomileusis, LASIK) were evaluated. Lacrimal fluid was collected before operation and in various terms after it in 28 patients (48 eyes) subjected to PRK and in 34 patients (59 eyes) subjected to LASIK. Biochemical analysis of the lacrimal fluid was performed on a mass spectrometer. After PRK the greatest changes in the protein spectrum occurred immediately after the operation before reepithelialization. Changes after LASIK were negligible. Refraction laser operations cause changes in the protein composition of tears, LASIK being more physiological than PRK from viewpoint of intactness of the protein composition of tears.

Detection of substance P in human tears by laser desorption mass spectrometry and immunoassay.

PURPOSE: To determine whether substance P is present in human tears. METHODS: Tear samples (1-2 microliters) were collected from one eye of each of 12 subjects. Two of the eyes had dry eye syndrome, two wore contact lenses and had dry eye syndrome, and eight were normal. Five of the eight normal eyes were scheduled to undergo excimer laser refractive surgery, and tears were collected from these eyes before and after surgery. Tear samples were analyzed by laser desorption mass spectrometry. Pooled samples from one individual were subjected to enzyme-linked immunoabsorbent assay. RESULTS: Laser desorption mass spectra of the 18 tear samples displayed well defined peaks with mass to charge (m/z) ratios ranging from 1343.7 to 1355.9 and/or 1356.9 to 1364.7, corresponding to an average m/z of 1349.8 +/- 1.13 for protonated substance P and 1361.2 +/- 0.54 for oxidized substance P obtained from 14 mass spectra of standards formulated with substance P concentrations ranging from 10(-4) M to 10(-12) M. As confirmation, an enzyme-linked immunoabsorbent assay performed twice on pooled tears from one eye detected substance P in both replicates at a concentration of 125 pg/ml (9.26 x 10(-11) M). CONCLUSIONS: These findings demonstrate that substance P is a component of tears obtained from normal eyes of men and women ranging in age from 26 to 60 years, from eyes fitted with contact lenses, from eyes with dry eye syndrome, and from eyes 1 and 2 days after excimer laser refractive surgery. Whether the concentration of substance P in tears varies with sex, age, or eye condition, the source of substance P in tears, and its role in tears remains to be discovered.

Analysis of rabbit tear fluid using capillary electrophoresis with UV or laser-induced fluorescence detection.

In preliminary studies of the development of tear analysis methodology that may eventually be useful in the clinical setting, the authors evaluated various protocols for analyzing rabbit tears by capillary zone electrophoresis (CZE). Conditions included the use of a 50-mM monosodium phosphate buffer, pH 2.5, or a 400-mM sodium borate buffer, pH 8.9, both with ultraviolet (UV) detection, as well as a 50-mM borate buffer, pH 8.5, with laser-induced fluorescence (LIF) detection of ATTO-TAG CBQ (Molecular Probes, Inc., Eugene, OR, U.S.A.) derivatized tears. All CZE analyses were performed with a P/ACE System 2100 instrument equipped with System Gold software (Beckman Instruments, Fullerton, CA, U.S.A.), using a 50 microns x 57 cm (50 cm to the window) fused-silica capillary, at 25 degrees C, with constant voltage of 20 kV for UV detection and 11 kV for LIF detection. Tear samples were collected from normal rabbit eyes by means of 10-microL glass micropipets. The volume of each sample was approximately 2 microL. Analysis using the phosphate buffer with UV detection produced as many as 35 peaks in each sample, of which 11 peaks were readily discerned. This compared favorably with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, which produced 32 bands with silver staining and 11 quantifiable bands with Coomassie brilliant blue staining. Many of the tear protein components have yet to be identified. CZE analysis with the high-ionic-strength borate buffer with UV detection produced only four peaks, and the low-ionic-strength borate buffer with LIF detection produced only six peaks. CZE analysis was completed in less than 1 hr, compared with 7-8 hr for SDS-PAGE. In summary, CZE analysis of tear fluid is comparable to CZE analysis of other bodily fluids and shows great potential for use in clinical diagnosis as well as for enhancing our understanding of the cellular actions of tears on the front of the eye.

Diclofenac sodium attenuates neural activity after photorefractive keratectomy in rabbits.

BACKGROUND: Photorefractive keratectomy for treating myopia has been associated with severe postoperative pain. The purpose of the present study was to investigate, in a rabbit model, the early effects of laser ablation on the physiology of the sensory nerves of the cornea. METHODS: Neurophysiologic monitoring of the corneal nerves was carried out 4 to 6 hours after 5-mm diameter, circular PRK ablations with a central depth of 85 microns. The effect of topical 0.1%, 0.25%, and 0.5% diclofenac sodium on the physiologic response to chemical and mechanical stimulation was evaluated. Corneas were prepared for anatomic studies of corneal innervation. RESULTS: Heightened neural activity was observed in the corneal nerves after ablation, and exaggerated responses to sensory stimulation with hypertonic sodium chloride solutions were recorded. Diclofenac sodium at 0.1% decreased the response to 0.5 M NaCl and 0.7 M NaCl to 67 +/- 9% and 68 +/- 7% (mean +/- standard error) of the control response. Mechanically sensitive nerves were more resistant to diclofenac; 0.1% diclofenac had no effect, but 0.25% diclofenac reduced the response significantly (P < 0.05). Anatomic observations showed that the ablation extended through the neural plexus. By 72 hours, long neurites tipped with growth cones had advanced into the injured region, indicating the initiation of neural regeneration. CONCLUSIONS: Diclofenac sodium was effective in attenuating neural activity in the cornea after excimer laser ablation. The action of this agent was shown to be different from that of a topical anesthetic, which rapidly and totally inhibited the response to all forms of stimulation.

Clinical analysis of excimer laser photorefractive keratectomy using a multiple zone technique for severe myopia.

PURPOSE: In an investigational procedure, excimer laser photorefractive keratectomy for severe myopia was performed at three clinical trial centers to determine the effectiveness of the multiple zone technique. METHODS: A VisX Model Twenty/Twenty excimer laser (VisX, Santa Clara, California) was used to perform photorefractive keratectomy on 14 severely myopic eyes (-10.37 to -24.5 diopters) of 12 patients by using a multiple zone technique. Postoperative follow-up ranged from six months to two years; retreatments were performed on four patients, with a follow-up of at least nine months. RESULTS: At six months postoperatively, before retreatment, three of the 14 eyes were within 2 diopters and seven of the 14 eyes were within 4 diopters of attempted correction. Regression of effect to more severe myopia was worse in five eyes treated with nitrogen gas blowing. Retreatments also demonstrated considerable myopic regression. Three patients had loss of two or more lines of best-corrected visual acuity, and these patients also had moderate or severe levels of haze. CONCLUSION: Excimer laser photorefractive keratectomy for severe myopia using a multiple zone technique is associated with considerable regression, haze, and loss of best-corrected visual actuity, especially when performed in association with nitrogen gas blowing.