The presence of sugar in the aqueous humor.
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Aqueous Humor↗
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[Clinical measurement of resistance to flow of aqueous humor].
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Aqueous Humor↗
Lysosomal enzyme activity in human aqueous humor.
Activity of the lysosomal enzymes N-acetyl-beta-D-hexosaminidase, alpha-D-mannosidase, beta-D-glucuronidase, and beta-D-galactosidase was detected in aqueous humor from eyes undergoing intraocular surgery. There was no correlation between lysosomal enzyme activity and age. Lysosomal enzyme activity in human released by ocular tissues surrounding the anterior chamber including the cornea, trabecular meshwork, ciliary body, iris, and lens. Their release into aqueous humor may have a role in regulating aqueous outflow in normal and glaucomatous eyes.
Adult↗
Non-invasive detection of antibiotics and physiological substances in the aqueous humor by Raman spectroscopy.
BACKGROUND AND OBJECTIVES: Laser Raman spectroscopy is an inelastic light scattering technique able to characterize molecules in aqueous environments. The purpose of this work is to develop a non-contact and non-invasive spectroscopic method to identify and eventually quantify the presence of medicines (e.g., antibiotics) and physiological substances (e.g., glucose) in the aqueous humor of the eye. STUDY DESIGN/MATERIALS AND METHODS: A new laser light delivery probe has been developed and adapted to a Raman spectroscopic system with the ability of favorable collection of the Raman light at 90 degrees scattering geometry while scanning the anterior chamber of the eye. Different amounts of ceftazidime, amphotericin B, and glucose had been injected in the aqueous humor of porcine eyes, maximum 24 hours after death and extraction, in-vitro. Raman measurements were excited with a visible (514.5 nm) laser beam at a power of 25 mW and an exposure/acquisition time of 1 second. RESULTS: The specific collection optics and Raman analysis components used in the present work have resolved the Raman signatures of probed molecules and low concentrations of ceftazidime (0.9 mg/mL), amphotericin B (9 microg/mL), and glucose (2 mg/ml) separately injected in the anterior chamber of porcine eyes were detected in vitro. CONCLUSION: This special illumination design gives the opportunity of avoiding the direct exposure to the laser light of basic cordial tissues of the eye, like lens and retina, although an optimum collection of scattered light is accomplished. Concentrations close to the minimum inhibitory concentration (MIC) have been detected for ceftazidime and amphotericin b; the detection of glucose has been realized at concentrations close to the early pathological levels of patients with diabetes.
Amphotericin B↗
The lens influences aqueous humor levels of transforming growth factor-beta 2.
BACKGROUND: Transforming growth factor-beta 2 (TGF-beta 2) is a pluripotent cytokine which has been suggested to play a number of roles in ocular physiologic and pathologic states. Intraocular fluid (i.o.f.) levels of TGF-beta 2 are quite high. Although the sources of ocular TGF-beta are not completely defined, the retinal pigment epithelium, the epithelium of the ciliary body and trabecular meshwork cells all secrete it. In this study we utilized canine lens and rabbit ciliary pigmented epithelial cell cultures to quantitate the in vitro secretion of TGF-beta 2. In addition, the effects of aphakia or the presence of cataractous lenses on IOF TGF-beta 2 levels were determined. METHODS: Lens and ciliary body epithelial cell culture supernatants and aqueous humors were assayed for total TGF-beta 2 levels by ELISA and bioassay. RESULTS: TGF-beta 2 accumulated in the media bathing lens epithelial cell cultures (0.7 +/- 0.03 ng/ml at day 2) and ciliary pigmented epithelial cell cultures (0.8 +/- 0.06 ng/ml at day 2) in a time-dependent manner. Surprisingly, aqueous humor from aphakic rabbit eyes contained significantly higher levels of TGF-beta 2 than their contralateral phakic controls. Furthermore, aqueous humor from canine eyes with cataracts also contained significantly higher levels of TGF-beta 2 than normal eyes. CONCLUSIONS: These results suggest that the lens secretes TGF-beta 2 and that the presence and status of the lens may influence IOF TGF-beta 2 levels.
Animals↗
Analysis of immunomodulatory activities of aqueous humor from eyes of mice with experimental autoimmune uveitis.
Aqueous humor (AqH) contains immunosuppressive factors, especially TGF-beta2, that contribute to the immune privileged status of the anterior chamber. However, this may not be true when the blood-ocular barrier is compromised by ocular inflammation. To determine the immunosuppressive status of AqH from murine eyes afflicted with experimental autoimmune uveitis, B10.A mice were immunized with interphotoreceptor retinoid-binding protein. AqH was collected from eyes of affected mice periodically after immunization and then evaluated for content of TGF-beta, proinflammatory cytokines, and the capacity to suppress anti-CD3-driven T cell proliferation. mRNA expression of selected cytokines in iris and ciliary body from inflamed eyes was analyzed by ribonuclease protection assay. We found that TGF-beta levels were significantly increased in AqH from EAU eyes on days 11, 17, and 28. AqH collected on day 11 (onset of disease) failed to suppress T cell proliferation and contained large amounts of locally produced IL-6 that antagonized TGF-beta. In contrast, AqH collected at 17 days (when ocular inflammation was progressively severe) re-expressed the ability to suppress T cell proliferation, in this case due to high levels of blood-derived TGF-beta1 and eye-derived TGF-beta2 in the absence of IL-6. Thus, during the onset of experimental autoimmune uveitis, the ocular microenvironment loses its immunosuppressive properties due to local production of IL-6. But as inflammation mounts, AqH IL-6 content falls, and the fluid reacquires sufficient TGF-beta eventually to suppress immunogenic inflammation. The paradoxical roles of IL-6 in antagonizing TGF-beta, while promoting TGF-beta accumulation during ocular inflammation, is discussed.
Adjuvants, Immunologic↗
Pilocarpine concentrations in aqueous humor following single drop application. I. Effect of soft contact lenses.
Following single drop instillation of pilocarpine, drug levels in individual aqueous humor samples of rabbits were determined by high-performance liquid chromatography and correlated with changes in pupillary diameter and intraocular pressure (IOP). Application of higher concentrations of pilocarpine resulted in higher drug levels but these differences were neither statistically significant nor consistently proportional to the strength applied. Maximum miosis occurred between 30 and 60 minutes and the maximum fall in IOP occurred between 1 and 2 hours. The influence of contact lenses with varying water content and thickness upon intraocular penetration of pilocarpine was studied. Contact lenses did not affect 30-minute concentrations of pilocarpine in aqueous humor. Although two-hour levels were significantly lower with contact lenses, miosis and IOP at these time periods were comparable with and without lenses. Neither water content nor thickness appreciably influenced miosis and IOP.
Animals↗
Effects of ion transport and channel-blocking drugs on aqueous humor formation in isolated bovine eye.
PURPOSE: To investigate the role of active chloride secretion by the ciliary epithelium in the formation of aqueous humor (AH), by using the in vitro perfused eye. METHODS: Bovine eyes collected from an abattoir were cannulated through the ophthalmic artery and perfused with oxygenated Krebs' solution at 37 degrees C. Aqueous humor formation (AHF) was measured by the fluorescein-dilution technique. Drugs were added to the perfusate and/or to the anterior chamber. RESULTS: NaK-adenosine triphosphatase (ATPase) inhibitor, ouabain (1.0 mM), produced a significant reduction in AHF by 46% and 42% when added to the stromal or aqueous side, respectively. When added to both sides (1.0 mM), it produced a reduction of 61%. Bumetanide (0.1 mM), a specific inhibitor of Na-K-2Cl cotransport, and furosemide (0.1 mM), a nonspecific anion transport inhibitor, produced 35% and 45% reductions when applied to the stromal side. DIDS (0.001-0.1 mM), which is believed to inhibit the Cl-HCO(3) exchanger, Na-HCO(3) cotransporter, and chloride channel, produced a dose-dependent reduction when added to the stromal side. The inhibition was 55% by the highest concentration used. 5-Nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB; 0.1 mM), a chloride channel blocker in the nonpigmented cells, produced a 25% reduction when applied to the aqueous side. Acetazolamide (0.1 mM), a carbonic anhydrase inhibitor, applied to the stromal side, produced 31% reduction. CONCLUSIONS: At least 60% of the AH is formed by active secretion in bovine eyes. Transport of anions through the ciliary epithelium (CE), particularly the chloride ion, plays a crucial role in AHF.
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
Adenosine 3',5'-monophosphate increases the outflow of aqueous humor from the rabbit eye.
Direct administration of cyclic-AMP into the anterior chamber increases the outflow facility of the eye for aqueous humor. This is consistent with the hypothesis that catecholamines lower the intraocular pressure of the rabbit eye, at least in part, by a cyclic-AMP-mediated mechanism. This mechanism is active in the outflow channels and increases the rate at which aqueous humor leaves the anterior chamber.
Animals↗
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.
Adrenergic alpha-Antagonists↗
Increase in aqueous humor production following D1 receptors activation by means of ibopamine.
BACKGROUND: Topically administered 2% ibopamine (a dopaminergic agonist) induces a transitory ocular hypertension in 92% of patients with primary open-angle glaucoma and in 52% of patients with normal tension glaucoma. In normal eyes, ibopamine has no effect on IOP. PURPOSE: The aim of the present study was to verify, by means of fluorophotometric techniques, which hydrodynamic changes could be induced in normal and glaucomatous eyes, stimulating the D1 receptor with 2% ibopamine administered topically. In addiction, we wanted to evaluate if ibopamine could modify IOP before and after an experimentally induced outflow system impairment in rabbits. METHODS: In study 1 we performed a measurement of aqueous humor flow in 6 healthy volunteers and in 6 glaucomatous patients, before and after 2% ibopamine administration. In study 2 the alteration of outflow pathways was induced by means of Laminaria Digitata in 10 rabbits. RESULTS: After 2% ibopamine administration we found a significant increase in aqueous humor production, both in glaucomatous (P = 0.035) and normal eyes (P = 0.004). In rabbits, we found no significant change in IOP at basal conditions. After experimentally induced outflow system impairment by laminaria, we observed a marked increase in IOP (+ 13.5 mmHg SD 7.2; P < = 0.001) following ibopamine administration. CONCLUSIONS: These experimental data have a diagnostic value in glaucoma, since they show how an intraocular hypertensive response due to ibopamine in normotensive eyes is a sign of initial outflow impairment. Moreover, the possibility to increase the aqueous humor production sets new trends in the treatment of post surgical ocular hypotony.
Administration, Topical↗
Detection of specific proteins in the aqueous humor in primary open-angle glaucoma.
To elucidate the mechanism of increased intraocular pressure in primary open-angle glaucoma (POAG), the protein profiles of aqueous humor obtained from POAG patients were compared with those of cataract patients as a control group. Aqueous humor proteins were analyzed by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and detected by the ultrasensitive silver staining technique. In 79% of the samples taken from POAG patients, protein bands of 140,000 or 160,000 daltons were stained, but none were stained from cataract patients. The presence of these protein bands revealed statistically significant differences between the two groups. Protein bands of 140,000 or 160,000 daltons were evenly visible at all ages in POAG patients, and the positivity of bands had no correlation with sex or initial intraocular pressure level. It is possible that the ultrastructural changes of the aqueous outflow pathway in POAG may be related to the changes in the aqueous protein, presence of 140,000 or 160,000 daltons protein bands.
Adult↗
Heterogeneity of lymphocytes from aqueous humor of patients with uveitis.
T lymphocyte subsets from inflammatory aqueous humor (n = 34) have shown one of three different patterns: group 1 = lower CD4+/CD8+ ratio in aqueous as compared to peripheral blood (0.43 +/- 0.25 vs. 1.96 +/- 0.82; n = 9); group 2 = similar CD4+/CD8+ ratio in aqueous and peripheral blood (1.76 +/- 0.60 vs. 1.60 +/- 0.62; n = 22); and group 3 = higher CD4+/CD8+ ratio in aqueous as compared to peripheral blood (7.53 +/- 0.47 vs. 2.13 +/- 0.55; n = 3). DR+ T lymphocytes were significantly higher, and natural killer cells significantly lower, in inflammatory aqueous as compared to peripheral blood.
Aqueous Humor↗
Transforming growth factor-beta levels in human aqueous humor of glaucomatous, diabetic and uveitic eyes.
PURPOSE: Transforming growth factor-beta2 is known to be present at elevated levels in the aqueous humor of patients with primary open angle glaucoma (POAG) and diabetes but not in uveitis-related secondary glaucoma. We investigated total TGF-beta2 levels and levels of the active form of TGF-beta2 in the aqueous humor of eyes with different types of glaucoma. METHODS: The concentration of the total and active form of TGF-beta2 was measured in 63 patients with primary open angle glaucoma, neovascular glaucoma complicated with diabetes (NVG), and secondary open angle glaucoma complicated with uveitis (SOAG) using a double antibody 'sandwich-indirect' ELISA method. RESULTS: The levels of total TGF-beta2 in the aqueous samples of POAG, NVG, and SOAG were elevated. The levels of active TGF-beta2 in the aqueous samples of POAG, and NVG were also elevated, whereas the level of active TGF-beta2 was within the normal range in the aqueous samples of SOAG. CONCLUSIONS: These results suggest that the level of TGF-beta2 may play a role in the pathology of various types of glaucoma.
Adult↗
Acute effect of thymoxamine on aqueous humor formation in the epinephrine-treated normal eye as measured by fluorophotometry.
The acute effect of 0.5% thymoxamine and 1% epinephrine on aqueous humor flow was evaluated by fluorophotometry in 25 normal human subjects in a randomized, double-blind, placebo-controlled study. Changes in intraocular pressure, anterior chamber volume, and pupil diameter were also measured. The results of this study indicate that epinephrine alone increases aqueous humor flow by 19% and that thymoxamine in a dose sufficient to produce miosis does not alter this acute effect of epinephrine.
Anterior Chamber↗
Aqueous humor dynamics in the aging human eye.
PURPOSE: Healthy subjects were recruited to identify normal, age-associated changes in intraocular pressure and aqueous humor dynamics. METHODS: Normal healthy subjects from two age groups were enrolled in the study: (1) those from 20 to 30 years of age (n = 51) and (2) those 60 years of age and older (n = 53). Intraocular pressure was measured by pneumatonometry, tonographic outflow facility by pneumatonography, and episcleral venous pressure by venomanometry. Aqueous flow and outflow facility were determined by a fluorophotometric technique. Uveoscleral outflow and anterior chamber volume were calculated. Results from the older group were compared with those from the younger group by means of unpaired, two-tailed t tests. RESULTS: Compared with the younger group, the older group showed significant differences as follows: smaller anterior chamber volume (160+/-39 vs. 247+/-39 microl; mean +/- SD; P< .00001), reduced aqueous flow (2.4+/-0.6 vs. 2.8+/-0.8 microl/minute; P = .002), and reduced uveoscleral outflow (1.10+/-0.81 vs. 1.52+/-0.81 microl/minute; P = .009). CONCLUSIONS: In the healthy aging eye, there is a reduction in the production of aqueous humor and a reduction in its drainage through the uveoscleral outflow pathway.
Adult↗
L-Arginine-nitric oxide pathway-related metabolites in the aqueous humor of diabetic patients.
PURPOSE: Nitric oxide (NO) is an important signal-transduction molecule that plays a significant role in the regulation of cardiovascular functions. In the L-arginine-NO pathway, NO synthase (NOS) converts L-arginine (L-Arg), the only known biologic substrate for NO formation, to NO and L-citrulline (L-Cit). Excessive NO production mediated by the inducible isoform of NOS has been implicated in the pathogenesis of various diseases. In the present study it was hypothesized that in vitreoretinal disorders such as diabetic retinopathy the production of L-Arg-NO pathway-related metabolites may be upregulated as a result of increased NO generation. METHODS: From 20 eyes of nondiabetic subjects and 22 eyes of diabetic patients with (n = 14) and without (n = 8) diabetic retinopathy, undiluted samples of aqueous humor were drawn before cataract surgery. Levels of L-Arg, L-Cit, and the specific NOS by-product N(G)-hydroxy-L-arginine (HOArg) were measured by high-performance liquid chromatography. RESULTS: L-Arg, L-Cit, and HOArg were detected in all aqueous humor samples from diabetic and nondiabetic patients (n = 42). Comparison of HOArg levels in nondiabetic and diabetic subjects showed significantly higher levels in diabetic patients (P = 0.002). Concentrations of HOArg were higher in samples from patients with (P = 0.005) and without diabetic retinopathy (P = 0.033) than in control subjects. No statistically significant differences were observed in L-Arg or L-Cit levels. CONCLUSIONS: Elevated levels of HOArg in the aqueous humor of diabetic patients reflect the possible role of NO as a significant factor in the regulation of retinal vascular functions and intraocular proliferative changes in diabetes mellitus in vivo. The control of intraocular NO production may constitute a potential therapeutic approach in diabetic retinopathy.
Aged↗
Comparison of protein mixtures in aqueous humor by membrane preconcentration - capillary electrophoresis - mass spectrometry.
The significance of proteomic research is coupled with the recent exponential growth of these investigations. Currently, the most popular techniques used for these studies include the coupling of 1- and 2-dimensional electrophoresis with mass spectrometric analysis of the extracted and digested proteins. However, detection limits of gel staining methods have led to a search for complimentary techniques that afford the detection of lower concentrations of biologically relevant proteins. In the present studies, we have evaluated the applicability of on-line capillary electrophoresis - mass spectrometry (CE-MS) for this application. Specifically, we used membrane preconcentration-CE-MS (mPC-CE-MS) to analyze 13 samples of human aqueous humor (AH) from patients with various ocular pathologies (cataract, cataract plus glaucoma, and cataract plus pseudoexfoliation syndrome). This approach enabled rapid analysis of a relatively large volume (1 microL of each specimen, and a protein map for each was created. Measured average molecular weights (Mr) were used to tentatively identify proteins after search of the SWISS-PROT database using TagIdent from ExPaSy. Among those proteins tentatively identified are beta-2 microglobulin (Mr 11731.2), apolipoprotein A1 (Mr 28078.6) and serum albumin (Mr 66400). Proteins with Mr of 4349 (unidentified), 11731.2 (beta-2 microglobulin), 13400-14100 (immunoglobulin fragments), 28078.2 (apolipoprotein A1) and approximately 68000 (serum albumin) were observed in the majority of specimens. Generally no significant differences were noted in the protein composition of aqueous humor samples from different pathologies. However, the absence of an Mr 13345 protein and its oxidized form (Mr 13361) in samples from patients with pseudoexfoliation syndrome was noted. Occasionally the alpha-and beta-chains of hemoglobin, a contaminant in aqueous humor introduced during sampling, were also detected. We conclude from these studies that mPC-CE-MS is an attractive complimentary technique for proteome research, as this approach enables direct mapping and characterization of low concentrations of proteins that are present in complex physiologically derived fluids.
Apolipoprotein A-I↗