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PubMed2005

A mathematical model for ocular tear and solute balance.

PURPOSE: In this paper we develop a mathematical model that can predict the steady-state tear film thickness and the dynamic tear film thickness and the solute concentration after instillation of a solute-laden fluid in the eye. METHODS: The mathematical model developed in this paper is based on a balance between the inflow and outflow of tears into the tear film. It incorporates a tear drainage model and a model that relates the tear film thickness to the meniscus radius of curvature. To predict the solute concentrations, the tear balance is coupled with the solute balance. The differential equations for the unsteady balances are solved numerically. RESULTS: The model predicts that the tear film thickness depends on a number of physiological factors, such as rates of tear production and evaporation, geometry and modulus of the canaliculi, and surface tension and viscosity of tears, and varies from about 3 to 15 microm. The model also predicts that the drainage time for an instilled volume of 15 microl is 1283 s. Additionally, the time required for the tracer concentration to decay to 1% of the value immediately after instillation of a drug-laden 40 microl drop is about 2480 s. Similarly, the time for intensity decay for a radioactive tracer after 25 microl instillation is about 1566 s. Also, the model predicts that the fraction of the instilled drug that reaches the cornea is about 1.3% for topical application of timolol. CONCLUSIONS: The predicted results agree reasonably with the reported experimental results, at least qualitatively. The model developed here can serve as a useful tool to develop a more precise understanding of various issues related to tears and also evaluate the effect of various parameters on the tear volume.

Eye↗
PubMed2000

Dry eye and closed eye tears.

PURPOSE: To establish accurate measurement of tear production in the diagnosis and monitoring of dry eye. METHODS: Reexamination of the Schirmer test indicated that the rate of tear secretion decreases with advancing age. We measured the noninvasive tear film break-up time (NIBUT) in normal and dry eye patients with our original apparatus. The NIBUT showed a reasonable level of accuracy with a cut-off value of 5 seconds. The lactoferrin concentration in tear fluids subsequently was found to have a cut-off value of 1.1 mg/mL with an optimal level of accuracy. RESULTS: The present findings suggest that a combination of clinical tests is needed for the diagnosis of dry eye syndrome. The closed eye condition induces subclinical inflammation on the ocular surface. Fibronectin, a high-molecular weight glycoprotein, plays an important role in corneal wound healing, although its concentration in various types of tear has not been investigated. Fibronectin concentration in open eye tears (19 +/- 24 ng/mL) was significantly different (p = 0.004) from that in closed eye tears (4127 +/- 3222 ng/mL). During reflex tearing with nasal stimulus, the concentration increased significantly (p = 0.028) after 100 mL of reflex tears (220 +/- 126 ng/mL). Albumin concentration in the same samples showed a pattern similar to that for fibronectin. Administration of a topical vasoconstrictor eliminated the increase in fibronectin concentration during reflex tearing. CONCLUSION: These findings suggest that fibronectin in the tear fluid was derived from plasma and that the increase in concentration in closed eye and reflex tear fluid was caused by leakage from dilated conjunctival blood vessels.

Adolescent↗
PubMed1995

Effect of age on tear osmolality.

Certain aspects of tear physiology change with age (reflex secretion and stability), whereas others remain unchanged (tear volume, evaporation rate, basal tear production). To establish the variation in tear osmolality with age, 0.2 to 0.4 microliter samples were collected from 50 normal males and 50 normal females (age range 17 to 75 years). Nanoliter aliquots of the samples were analyzed by freezing point depression nanoliter osmometry. Mean tear osmolality for the whole group was 303.6 +/- 13.0 mOsm/kg and was not significantly affected by age (r2 = 0.014, p = 0.378). For males the mean was 307.1 +/- 14.4 and for females 300.1 +/- 10.4 mOsm/kg, a significant gender difference (p = 0.006). Males under age 41 had tear osmolality which was not significantly different from that of males over 41 years of age (307.7 +/- 15.6 and 305.7 +/- 11.5 mOsm/kg, respectively, p = 0.671) but the mean tear osmolality for females under age 41 years (297.6 +/- 11.2 mOsm/kg) was significantly different from that of older females (304.8 +/- 6.7 mOsm/kg, p = 0.017). There was no significant correlation between age and osmolality for males (r2 = 0.003, p = 0.699), but there was a significant correlation for females (r2 = 0.159, p = 0.004). Males under age 41 years have a significantly higher mean tear osmolality than females under 41 years of age (p = 0.003). The age and gender differences found for tear osmolality can be accounted for by the low values in young females, which may be related to the increased tear flow observed in young females. A reduction in this high flow rate to a more "normal" level in older females may account for the rise in tear osmolality to a level which is not significantly different from that of older males. Generally in normal eyes, the osmolality of tears remains within normal limits and is not affected by age.

Adolescent↗
PubMed1997

Tear fluid plasmin activity of dry eye patients with Sjögren's syndrome.

Thirty-two eyes of 16 patients with verified Sjögren's syndrome were examined for clinical signs of dry eye. Tear fluid samples were collected for plasmin assay. Ophthalmologic examinations included estimation of conjunctival or corneal discharge, filament formation and presence of conjunctival or corneal epithelial defects, assessment of tear meniscus height and measurement of tear fluid break-up time, Schirmer test, and fluorescein and Rose-Bengal staining graded by the van Bijsterveld score. Tear fluid plasmin activity (IU/l) was determined by a fluorometric assay and tear fluid flow (microl/min) was measured for calculation of tear fluid plasmin activity release (microIU/min). All patients had relatively dry eyes; the mean Schirmer test value was 5.7 +/- 0.5 mm/5 min. The mean tear fluid break-up time was also low, 7.7 +/- 0.5 s. The mean Bijsterveld score value was 2.5 +/- 0.5. Because collection of tear fluid by microcapillaries for the plasmin assay was difficult due to the low tear fluid flow rate, it was necessary to drop 20 microl of balanced salt solution topically on the cornea to aspirate a tear fluid sample. Despite this, the mean tear fluid plasmin activity was higher than in control individuals (7.75 +/- 1.51 IU/l vs. 0.73, range 0.64-0.80 IU/l). On the basis of these findings we conclude that elevated tear fluid proteolytic activity may play a role in the pathology of dry eye/ocular surface disease.

Adult↗
PubMed1986

The concentration of lactoferrin in tears of normals and of diabetics.

The tear lactoferrin content and the tear secretion rate was determined in 105 normal subjects (age range: 15-93 years) and 21 diabetic patients (age 21-78 years). Tear sampling was performed using glass capillaries and the lactoferrin concentration in tears was tested employing an enzyme-linked immunosorbent assay. Tear secretion rate was measured using standardized Halberg filter paper placed in the inferior fornix for 5 min. The lactoferrin level of normal tears showed a significant inverse correlation with age, the mean value of the 8th decade (0.81 mg/ml) constituting only about half the value of the lactoferrin level found in the 3rd decade (1.48 mg/ml). The tear secretion rate also displayed a significant decrease with age, but no correlation could be demonstrated between the tear secretion rate and lactoferrin concentration. No difference was found in lactoferrin level between men and women. We were not able to demonstrate any correlation between lactoferrin content of tears and the duration of diabetes in the 21 diabetic subjects, and there was no difference between the level of lactoferrin in normal tears compared to the concentration in diabetic tears. No difference was found between the tear secretion rate in normals and in diabetics.

Adolescent↗
PubMed1987

Tear and serum antibody response to Chlamydia trachomatis antigens during acute chlamydial conjunctivitis in monkeys as determined by immunoblotting.

In this study, we examined the temporal antibody response by immunoblotting analysis in tears and sera of three cynomolgus monkeys (Macaca fasicularis) with primary acute Chlamydia trachomatis serovar B conjunctivitis. The objective was to identify chlamydial antigens stimulating antibody during the host responses in the course of this self-limiting infection with the rationale that they may be protective antigens. The major outer membrane protein (MOMP), lipopolysaccharide (LPS), and polypeptides of 60 and 68 kilodaltons (kDa) were the predominant antigens recognized by immunoglobulin A (IgA) in monkey tears. Tear IgA antibody specific for the MOMP was first detected 14 days postinfection, whereas tear IgA reactive with LPS or the 68- and 60-kDa polypeptides was first detectable on day 21. Tear IgA antibodies specific for each of these antigens persisted in tears through day 56, 4 weeks after both peak clinical disease and recovery of the organism from the conjunctivae. In contrast, tear IgG antibodies peaked at approximately 28 days postinfection, the time of maximal inflammatory response. The IgG response in monkey sera was similar to that observed for tear antibodies, in that the MOMP, 60-, and 68-kDa polypeptides were the primary immunogens. The exception was that IgG antibody against these antigens was detected 1 week later than that observed for tear IgA antibodies. Of three monkeys that responded with tear IgA antibody against LPS, one did not have detectable serum IgG LPS antibody. The specificity of the tear IgA antibody response of monkeys was determined by immunoblotting nine other C. trachomatis serovars in addition to the homologous B serovar. The tear IgA response to the MOMP was predominantly B complex subspecies-specific (serovars B, Ba, D, and E), whereas the response to chlamydial LPS was found to be species-specific. The significance of these observations in relation to previous vaccine studies in nonhuman primates is discussed.

Animals↗
PubMed2003

Tear evaporation dynamics in normal subjects and subjects with obstructive meibomian gland dysfunction.

PURPOSE: To test a newly developed tear evaporimetry system that detects real-time changes in tear evaporation rates and shows the tear film stability in patients with obstructive meibomian gland dysfunction (MGD). METHODS: A ventilated chamber system with high-sensitivity microbalance sensor was used to evaluate tear evaporation. Tear evaporation rates and dynamic changes in them in response to blinking ("flip heights") were measured. Both were compared in 38 eyes of 22 normal subjects and 32 eyes of 21 patients with obstructive MGD, in a prospective case-control study. The relationship between tear evaporation rates and flip heights to meibomian gland orifice obstruction was also analyzed. RESULTS: Changes in tear evaporation rates produced by blinking were detected. The tear evaporation rates in the patients' group were 5.8 +/- 2.7(10(-7)) g/cm(2) per second, significantly higher than in normal subjects (4.1 +/- 1.4[10(-7)] g/cm(2) per second; P = 0.0008). The flip heights in the obstructive MGD group were 0.58 +/- 0.33(10(-7)) g/cm(2) per second, significantly higher than in normal subjects (0.39 +/- 0.27[10(-7)] g/cm(2) per second, P = 0.02). The correlations between both tear evaporation rates and flip heights to the meibomian gland orifice obstruction score were statistically significant (P < 0.0001 and P = 0.004, respectively). CONCLUSIONS: This new system was helpful in differentiating MGD patients from normal subjects. These significantly higher evaporation rates and higher flip heights reflect the unstable tear evaporation and may well indicate unstable tear film in patients with obstructive MGD with abnormal evaporative tear loss.

Adult↗
PubMed2003

Computer-synthesis of an interference color chart of human tear lipid layer, by a colorimetric approach.

PURPOSE: To synthesize an interference color chart for the specific tear lipid layer interference camera, DR-1, for the conversion of the tear film lipid layer thickness into color graphic information--that is, for the quantification of the tear interference image--by a colorimetric approach. METHODS: Because the color of the tear lipid layer interference image is visualized by a white light source interference phenomena, to produce gamma-corrected red, green, and blue (RGB) values of a specific interference color at a certain tear lipid film thickness, XYZ tristimulus values of the Commission Internationale de l'Eclairage (CIE) were obtained. XYZ tristimulus values were calculated from the light source spectrum of the DR-1 camera, the color-matching function of CIE, and the reflectance of the tear interference image in wavelengths ranging from 380 to 780 nm. These calculated interference colors were synthesized ranging from 0 nm to 1000 nm of lipid film thickness to produce a color chart. The applicability of the new color chart in the analysis of the lipid layer thickness was tested on a healthy control subject with normal tear function and a patient with dry eye who had aqueous tear deficiency and meibomian gland obstruction. RESULTS: The specific tear interference color chart for the DR-1 camera was obtained with RGB and XYZ tristimulus values. The interference chart ranged from 0 to approximately the 5th interference order. The interference colors from clinical DR-1 images could be converted to lipid thickness data by using the color chart system. CONCLUSIONS: A new tear interference color chart was developed in this study, which may be of benefit in converting tear interference color information to data describing the thickness of the tear film lipid layer.

Adult↗
PubMed2005

Tear lipocalin: evidence for a scavenging function to remove lipids from the human corneal surface.

PURPOSE: Lipid contamination of the cornea may create an unwettable surface and result in desiccation of the corneal epithelium. Tear lipocalin (TL), also known as lipocalin-1, is the principal lipid-binding protein in tears. TL has been shown to scavenge lipids from hydrophobic surfaces. The hypothesis that TL can remove contaminating fatty acids and phospholipids from the human corneal surface was tested. METHODS: TL was purified from pooled human tear samples by size exclusion and ion exchange chromatographies. Tears depleted of TL were reconstituted from fractions eluted by size exclusion chromatography that did not contain TL. Fresh and formalin-fixed human corneas were obtained from exenteration specimens. Fluorescent analogs of either palmitic acid or phosphatidylcholine were applied to the corneal epithelial surface. Tears, TL, or tears depleted of TL were applied over the corneas, and spectrofluorometry and fluorescent stereomicroscopy were used to monitor the removal of fluorescent lipids. Tears used in the experiments were then fractionated by size exclusion chromatography to determine the component of tears associated with fluorescent lipids. RESULTS: Significant enhancement of fluorescence for 16AP and NBD C(6)-HPC was evident in solutions incubated with whole tears and purified TL but not with tears depleted of TL for fixed and unfixed corneas. After the experiment, size exclusion fractions of tears showed that the fluorescence component coeluted with TL. CONCLUSIONS: TL scavenges lipids from the human corneal surface and delivers them into the aqueous phase of tears. TL may have an important role in removing lipids from the corneal surface to maintain the wettability and integrity of the ocular surface.

Carrier Proteins↗
PubMed2000

Comparison of tear proteins of llamas and cattle.

OBJECTIVE: To analyze and compare contents of the preocular tear films of llamas and cattle. ANIMALS: 40 llamas and 35 cattle. PROCEDURE: Tear pH was determined by use of a pH meter. Total protein concentration was determined by use of 2 microtiter methods. Tear proteins were separated by use of electrophoresis and molecular weights of bands were calculated. Western blot immunoassay was used to detect IgA, lactoferrin, transferrin, ceruloplasmin, alpha1-antitrypsin, alpha1-amylase, and alpha2-macroglobulin. Enzyme electrophoresis was used to detect proteases. RESULTS: The pH of llama and cattle tears were 8.05 +/- 0.01 and 8.10 +/- 0.01, respectively. For results of both methods, total protein concentration of llama tears was significantly greater than that of cattle tears. Molecular weights of tear protein bands were similar within and between the 2 species, although llama tears had a distinct 13.6-kd band that was not detected in cattle. Lactoferrin, IgA, transferrin, ceruloplasmin, alpha1-antitrypsin, alpha1-amylase, alpha2-macroglobulin, and proteases were detected in both species. CONCLUSIONS AND CLINICAL RELEVANCE: Llama tears have significantly greater total protein concentration than cattle tears, whereas pH is similar between species. Because little variation was detected within species for the number and molecular weight of protein bands, pooling of tears for analysis is justified. Results suggest that lactoferrin, ceruloplasmin, transferrin, alpha1-antitrypsin, alpha2-macroglobulin, alpha1-amylase, and IgA are present in the tears of llamas and cattle.

Amino Acids↗
PubMedDate not supplied

Tear fluid cellular fibronectin levels after photorefractive keratectomy.

BACKGROUND: Fibronectin is supposed to have an important role in wound healing. The extradomain A-containing cellular fibronectin (EDAcFn) refers to fibronectin, which instead of being a hepatocyte derived component of blood plasma or body fluids, is produced locally. The present study was undertaken to clarify the possible changes in excretion of EDAcFn in tears following excimer laser photorefractive keratectomy (PRK). METHODS: An immunoassay was used to determine EDAcFn concentrations in human tear fluid samples of 11 eyes after PRK. Tear fluids were collected with scaled microcapillaries preoperatively as well as 1, 2, and 7 days after PRK. The time used to collect a known volume of tears was registered. This was done to estimate the dilution effect related to the hypersecretion of tears after PRK. RESULTS: The mean preoperative tear fluid EDAcFn concentration was 0.28 +/- 0.07 ng/microliter with a wide range (0.05 to 0.63). The tear fluid flow-corrected excretion of EDAcFn was 1.36 +/- 0.35 ng/min (range, 0.145 to 3.50). There was a significant increase in both postoperative tear fluid flow and excretion of EDAcFn on days 1 and 2. The elevation of the mean EDAcFn concentration did not decrease in spite of reflex tearing. The mean excretion of EDAcFn in tears was 28-fold on the first and 17-fold on the second postoperative day. Normal level was reached by day 7. CONCLUSION: There is a rapid increase in excretion of EDAcFn in tears following PRK. This seems to last only as long as an epithelial defect persists. The epithelium of all eyes healed in 3 to 4 days in spite of wide interindividual variations in both tear fluid flow and EDAcFn excretion.

Adult↗
PubMed2001

Pro- and anti-inflammatory forms of interleukin-1 in the tear fluid and conjunctiva of patients with dry-eye disease.

PURPOSE: To compare the expression of the pro- and anti-inflammatory forms of interleukin (IL)-1 in the tear fluid and conjunctival epithelium of normal eyes and those with dry-eye disease. METHODS: The concentrations of IL-1 alpha, IL-1 beta (precursor and mature forms), and IL-1 receptor antagonist (IL-1Ra) were measured by ELISA in tear fluid samples obtained from normal individuals and patients with dry eye who had rosacea-associated meibomian gland disease (MGD) or Sjögren's syndrome (SS) aqueous tear deficiency (ATD). These cytokines were also measured in normal tear fluid before and after nasal stimulation to induce reflex tearing. The relative expression of these cytokines was evaluated in conjunctival impression cytology specimens and conjunctival biopsy tissue obtained from normal subjects and SS ATD-affected patients using immunofluorescent staining. Matrix metalloproteinase (MMP)-9 concentration and activity in the tear fluid were evaluated with gelatin zymography and with an MMP-9 activity assay kit, respectively. RESULTS: Compared with normal subjects, the concentration of IL-1 alpha and mature IL-1 beta in the tear fluid was increased, and the concentration of precursor IL-1 beta was decreased in patients with MGD (P < 0.05, P = 0.02, and P < 0.01, respectively) and SS ATD (P < 0.001, P = 0.02, and P < 0.001, respectively). There was no significant change in the concentration of IL-1 alpha, precursor IL-1 beta, and IL-1Ra in reflex tear fluid, indicating that the lacrimal glands may secrete these cytokines. The activity of MMP-9, a physiological activator of IL-1 beta, was significantly elevated in the tear fluid of both dry-eye groups compared with normal subjects. A strong positive correlation was observed between the intensity of corneal fluorescein staining and the tear fluid IL-1 alpha concentration (r(2) = 0.17, P < 0.02) and the mature-to-precursor IL-1 beta ratio (r(2) = 0.46, P < 0.001). Positive immunofluorescent staining for IL-1 alpha, mature IL-1 beta, and IL-1Ra was observed in a significantly greater percentage of conjunctival cytology specimens from eyes with SS ATD than in those from normal eyes (P < 0.01 for IL-1 alpha, P < 0.009 for mature IL-1 beta, and P < 0.05 for IL-1Ra). CONCLUSIONS: Dry-eye disease is accompanied by an increase in the proinflammatory forms of IL-1 (IL-1 alpha and mature IL-1 beta) and a decrease in the biologically inactive precursor IL-1 beta in tear fluid. Increased protease activity on the ocular surface may be one mechanism by which precursor IL-1 beta is cleaved to the mature, biologically active form. The conjunctival epithelium appears to be one source of the increased concentration of IL-1 in the tear fluid of patients with dry-eye disease. These results suggest that IL-1 may play a key role in the pathogenesis of keratoconjunctivitis sicca.

Adult↗
PubMed2001

Phospholipase A(2) in rabbit tears: a host defense against Staphylococcus aureus.

PURPOSE: This study analyzed rabbit tears for anti-staphylococcal activity, the role of phospholipase A(2) (PLA2) in this reaction, and the ability of enzyme inhibitors to promote bacterial survival. METHODS: Contact lenses with Staphylococcus aureus were applied to scarified rabbit eyes. The colony-forming units (CFU) per cornea or lens were determined and pathology was scored by slit-lamp examination (SLE). The bactericidal activity was measured by incubating bacteria with rabbit tears or PLA2 at 33 degrees or 37 degrees C. Radiolabeled S. aureus was incubated with PLA2 or tears to quantify the release of a membrane component that was identified by thin-layer chromatography. Inhibitors of these reactions were also analyzed. RESULTS: Application of Staphylococcus, on contact lenses, to rabbit corneas resulted in bacterial killing and limited inflammation. Incubation of tears and bacteria (1:1; v/v) in tryptic soy broth at 33 degrees C decreased CFU approximately 4 logs. Tears (> or =30 microl) or PLA2 (> or =30 U) incubated with bacteria in phosphate-buffered saline were bactericidal. PLA2 (> or =0.2 U) or tears (> or =2 microl) cleaved bacterial membranes, liberating arachidonic acid. Spermidine or tetracaine inhibited cleavage of bacterial membranes by tears or PLA2 and spermidine promoted bacterial survival and growth in tears. Tears (60 microl) killed >99% of the bacterial inoculum, whereas bacteria incubated in tears plus spermidine approximately doubled in number. CONCLUSIONS: PLA2 in rabbit tears kills Staphylococcus by hydrolyzing bacterial membranes to release arachidonic acid. Spermidine and tetracaine inhibited PLA2 activity and spermidine protected Staphylococcus from PLA2 in rabbit tears.

Animals↗
PubMed2001

Water-soluble antioxidants in human tears: effect of the collection method.

PURPOSE: To resolve differences in published data on tear antioxidant levels by comparing the concentration of water-soluble antioxidants in human reflex tears collected by capillary tube and by the Schirmer strip collection method and in basal and reflex tears collected using the Schirmer strip method. METHODS: Yawn-induced reflex tears (collected simultaneously by capillary tubes and by Schirmer strips) and basal tears (by Schirmer strips and using local anesthetic) were collected from 12 healthy subjects. Tear cysteine, ascorbate, glutathione, urate, and tyrosine were measured by high-performance liquid chromatography within a few minutes of collection. RESULTS: Cysteine, ascorbate, glutathione, and tyrosine were 5 to 10 times higher (P < 0.01) in both reflex and basal tears collected by Schirmer strip compared with reflex tears collected by capillary tube from the same subject. Urate levels were slightly but nonsignificantly higher in Schirmer strip samples (P > 0.05). CONCLUSIONS: The conflict in published data on tear antioxidants is caused by differences in collection methods. With the exception of urate, antioxidants accumulate to very high levels in corneal cells. Spuriously high antioxidant levels in tears collected using Schirmer strips, therefore, are most probably caused by contamination with intracellular constituents. The capillary tube collection method is proposed as the method of choice for reflex tear collection for biochemical studies. This less-invasive method facilitates the evaluation of tear antioxidant levels as a biomonitoring tool for corneal health. Although moderately increased antioxidant levels may be beneficial, the authors hypothesize that marked increases may indicate damage to the ocular surface.

Adult↗
PubMed1991

Changes in human tear protein levels with progressively increasing stimulus.

The levels of 13 proteins were measured in six tear samples collected atraumatically at progressively increasing flow rate from nonstimulated (less than 0.5 microliter/min) to highly stimulated (greater than 50 microliters/min) in ten subjects. Tears were fractionated initially by size-exclusion high-performance liquid chromatography (SE-HPLC). Enzyme-linked immunosorbent assays and kinetic assays were then applied to relevant SE-HPLC fractions to determine specific protein levels. Nine of the 13 proteins assayed showed significantly higher concentrations in nonstimulated tears than in any other tear sample. Immunoglobulin (Ig) M, secretory IgA, polymeric IgA1, and polymeric IgA2 all decreased progressively in concentration from nonstimulated tears to the higher flow-rate stimulated samples. The level of IgG, albumin, and transferrin showed a large drop in concentration between nonstimulated tears and the first (lowest flow-rate) stimulated sample, with relatively little decrease for any subsequent sample. Levels of lactoferrin, tear-specific prealbumin, lysozyme, and peroxidase were relatively constant throughout the series of tear samples. These results indicate that the mechanisms responsible for changes in concentration of constitutive, serum-derived, and regulated tear proteins with stimulus can be studied successfully using noninvasive methods to collect human tears. They also show that simply distinguishing between nonstimulated and stimulated tears is not sufficient to completely characterize the effect of stimulus conditions on tear protein composition.

Albumins↗
PubMed1984

Hormonal influence on the secretory immune system of the eye: androgen regulation of secretory component levels in rat tears.

The present study examined the influence of gender and steroid hormones on the level of free secretory component (SC) in tears of rats. The SC concentration in tears of male rats was approximately five-fold greater than that found in tears of females. This difference could not be accounted for by variations in tear volume or a suppression of tear SC content in female rats during certain stages of the estrous cycle. Rather, the elevated level of SC in tears of males appeared to be due to the effect of androgens. Castration of male, but not female, rats resulted in a time-dependent decrease in the SC content of tears. Administration of testosterone to orchiectomized rats reversed this decline and induced a time- and dose-dependent increase in the tear SC concentration. This response appeared specific for androgens, because treatment of orchiectomized rats with progesterone, estradiol, or cortisol had no effect on total tear SC. Consistent with this hypothesis was our finding that injection of castrated male rats with 5 alpha-dihydrotestosterone also raised the tear SC concentration. Of interest, the androgen-induced increase of the SC level in tears was not accompanied by similar changes in total tear protein. These results suggest that androgens may influence the production and/or secretion of SC by ocular tissues. Furthermore, our findings indicate that androgens play a role in the ocular secretory immune system of the rat.

Androgens↗
PubMed1994

Tear fluid plasmin activity after excimer laser photorefractive keratectomy.

PURPOSE: Elevated tear fluid plasmin activity may correlate with delayed healing of corneal wounds. The present study was performed to establish the tear fluid plasmin activity after photorefractive keratoablation (PRK). METHODS: Tear fluid aspirated with microcapillaries was subjected to a fluorometric plasmin assay using the 7-amido-4-trifluoromethylcoumarin derivate of the tripeptide H-D-Val-Leu-Lys as substrate. RESULTS: Tear fluid flow, plasmin activity, and flow-corrected plasmin excretion rate in tears (plasmin flux) were determined preoperatively and 1, 2, and 7 days after PRK. The preoperative tear fluid flow was 6.55 microliters/min (median; range, 1.8 to 21.8 microliters/min), plasmin activity was 1.29 IU/l (median; range, 0.6 to 6.9 IU/l), and the excretion of plasmin in tears was 11.7 microIU/min (median; range, 1.6 to 41.5 microIU). A statistically significant decrease in tear fluid plasmin activity was found during the follow-up period on the first (0.6 IU/l; range, 0.6 to 1.7 IU/l, P < 0.01) and second (0.65 IU/l; range, 0.6 to 1.49 IU/l, P < 0.01) postoperative days. On the other hand, significant elevation of both tear fluid flow and plasmin flux values occurred during the first two postoperative days. The median plasmin flux values on days 1, 2, and 7 were 57.35 microIU/min (range, 16 to 540 microIU/min, P < 0.01), 40.0 microIU/min (range, 13.3 to 222.8 microIU/min, P < 0.01), and 10.2 microIU/min (range, 2.2 to 90.7 microIU/min, P > 0.05), respectively. CONCLUSION: The marked elevation of tear fluid flow coincided with the persistence of an epithelial defect. However, because of the acceleration of tear fluid flow, proteolytic activity due to plasmin (IU/l) actually decreases. Consequently, the increased excretion of plasmin in tears (plasmin flux) does not lead to highly elevated plasmin activity, which could inhibit wound healing. It seems to be a natural healing response because all corneas were epithelialized normally by or on day 3.

Cornea↗
PubMed1996

Direct noninvasive measurement of tear oxygen tension beneath gas-permeable contact lenses in rabbits.

PURPOSE: To develop a noninvasive measurement of tear oxygen tension and to determine tear oxygen tension and calculated oxygen flux rates during wear of contact lenses of varying oxygen transmissibility (Dk/L). METHODS: The basis of the tear oxygen tension (PO2) measurement is phosphorescence quenching of Pd-meso-tetra-(4-carboxyphenyl) porphine by oxygen. A fiber-optic bundle, positioned approximately 2 mm from a cuvette or from the corneal surface, delivers an excitation flash (< 4 microseconds, 539 +/- 23 nm) and collects the phosphorescence emission (> 645 nm) of the dye at 1 MHz for 2 ms. Porphine (500 microM) + 1% albumin is instilled directly into the tears of a sedated rabbit, and phosphorescence is measured before and after contact lens insertion. RESULTS: Phosphorescence intensity decay lifetimes are related inversely to PO2 and follow the Stern-Volmer relationship. Calibration control experiments revealed a temperature effect on lifetime of 1% per degree, dye concentration independence over a 100-fold dilution, and no significant pH effect between 6.8 and 7.9. Phosphorescence lifetimes indicated that tear PO2 beneath contact lenses reached steady state within 3 to 8 minutes. Mean steady state tear PO2 (mm Hg +/- SE; N > 7) beneath contact lenses was: polymethylmethacrylate (Dk/L = 0), tear PO2 = 0.58 +/- 0.22; Oxyflow f30 (Dk/L = 15.3), tear PO2 = 27.7 +/- 3.4; Fluorex 700 Dk/L = 30), tear PO2 = 52.5 +/- 3.8; Oxyflow 151 (Dk/L = 51.9), tear PO2 = 78.7 +/- 23.9. Steady state tear PO2 beneath an Oxyflow f30 after application of topical anesthetic and 10 minutes of lens wear was 41 +/- 1.2 mm Hg, significantly higher than without anesthetic. CONCLUSIONS: Phosphorescence-based measurements allow the direct, sensitive, and noninvasive assessment of oxygen availability beneath a contact lens. Oxygen levels beneath permeable contact lenses found by direct measurement are significantly lower than what mathematical models predict. The measured values, however, are close to PO2 estimates using the equivalent oxygen percentage (EOP) technique. Tear PO2 measurements may be used to examine metabolic disturbances as shown by reduced oxygen consumption rates after application of topical anesthetic.

Anesthetics, Local↗