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Results for “Aqueous Humor”

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PubMed1998

The effect of antiglaucoma drugs on rabbit aqueous humor proteins determined by gel electrophoresis.

Previous studies indicate that there may be differences in the protein composition of the aqueous humor in normal and glaucomatous human eyes. These differences in protein composition and concentration may be due to the topical antiglaucoma medications used to treat glaucoma. These differences should be distinguished from any possible protein composition changes due to glaucoma. In order to study the effects of topical antiglaucoma medications on aqueous humor protein composition, we analyzed the aqueous humor of rabbit eyes topically treated with various antiglaucoma medications (timolol, pilocarpine, and dipivefrin). One eye of each rabbit was treated with the experimental drug, and the fellow eye was treated with saline solution (control). Thirty-six aqueous humor samples from 18 rabbits were obtained after 24 hours and 36 samples were obtained from 18 additional rabbits after 7 days of topical drug treatment. The protein composition of the aqueous humor samples was analyzed by sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Significant protein differences in aqueous humor samples were found between the eyes treated with timolol and the control eyes, whereas no significant differences were found between the eyes treated with pilocarpine or dipivefrin and the controls. These protein differences ranged from 14,000 to 18,000 daltons in molecular weight.

Administration, Topical↗

PubMed1992

Aqueous humor hydrogen peroxide analysis with dichlorophenol-indophenol.

Hydrogen peroxide is now reported to be a normal aqueous humor component present, in the low microM concentration range, in the animal species which have been studied. This finding was established with the exclusive use of the dichlorophenol-indophenol method of analysis. In this procedure, aqueous humor is added to a blue, oxidized dichlorophenol-indophenol solution. The 605 nm absorbance of this solution immediately decreases in response to the reducing action of ascorbate present in the sample. The extent of reoxidation of the solution upon the addition of peroxidase, as measured by the increase in its 605 nm absorbance, can be quantitatively related to the concentration of H2O2 in the sample. A close examination of this method revealed that reduced dichlorophenol-indophenol spontaneously reoxidizes at a rate of 0.03 nmol min-1 microM-1, with generation of H2O2. H2O2 generation was unequivocally established by analysis of the temporal dependency of the absorbance increase produced by peroxidase in the absence of added H2O2 and by the sensitivity of this phenomenon to catalase. This spontaneous production of H2O2, on the other hand, cannot be attributed to ascorbate auto-oxidation because added ascorbate quantitatively reacts with dichlorophenol-indophenol, provided that an excess of the latter is maintained. This method then has an enormous potential to overestimate H2O2 in any sample. On the other hand, the response of the assay system to a given level of H2O2 depends on the level of reduction previously produced by ascorbate. This results in an artifactual positive correlation between ascorbate and H2O2 levels in samples containing variable amounts of ascorbate. In spite of these serious limitations the method can still be useful to measure H2O2 if appropriate precautions are taken. When using it for the analysis of rabbit aqueous humor H2O2 without correcting for the H2O2 generated during the assay and ignoring differences in the level of ascorbate in the samples, we obtained an average value of 25.3 microM H2O2, which coincides with that reported in the literature for the rabbit, but is obviously incorrect. When analysing aqueous humor there was the additional variable of the aqueous humor itself inhibiting the rate of dichlorophenol-indophenol auto-oxidation and so the final, corrected figure for H2O2 concentration in the aqueous humor became uncertain, since the auto-oxidation of the substrate could not be properly subtracted.(ABSTRACT TRUNCATED AT 400 WORDS)

2,6-Dichloroindophenol↗

PubMed2001

Profile of cytokines in aqueous humor from corneal graft recipients.

AIM: Due to the immune privilege of the anterior eye chamber, the success rate of corneal transplantation can reach 90%. The aim of this study was to determine cytokine profile in aqueous humor of patients undergoing corneal transplantation, and to establish whether cytokine profile at the time of surgery influenced corneal graft outcome. METHODS: Proinflammatory (TNF-beta and IFN-gamma) and immunosuppressive (TGF-beta2) cytokine levels were measured in aqueous humor and serum of 44 patients. Non-inflammatory corneal diseases included keratoconus (n=8), bullous keratopathy (n=7), and stromal dystrophy (n=3). Inflammatory diseases included corneal scars (n=10), graft rejection (n=5), pending perforation (n=4), chemical burns (n=4), rejection/ uveitis (n=1), infectious keratitis (n=1), and perforated ulcer (n=1). Control aqueous humor and sera were obtained from cadavers without corneal pathology. RESULTS: The concentration of TGFbeta2 in the aqueous humor in non-inflammatory corneal diseases was similar to that of controls (2,605+/-204 pg/mL vs 2,200+/-100 pg/mL). In inflammatory corneal diseases, the concentration of TGFbeta2 in aqueous humor was significantly lower (1,400+/-375 pg/mL, p<0.001). TNF-beta was detected in the aqueous humor of 16 out of 26 patients with inflammatory corneal diseases and in all patients with stromal dystrophies, but was undetectable in cases of keratoconus and bullous keratopathy. Low levels of IFN-gamma were present in all aqueous humor samples. Patients' sera contained significantly less cytokine (up to 252 pg/mL) then their aqueous humor (p<0.001). We have set an arbitrary cut-off point for TGF-beta2 level in aqueous humor at 1,500 pg/mL and divided all investigated samples (from 44 patients and 10 controls) into two groups, one with high and the other with low TGF-beta2 concentration. The coefficient of contingency showed that patients with high TGF-beta2 concentration in their aqueous humor had significantly greater chance for graft acceptance than those with low TGF-beta2 concentration (p<0.001). CONCLUSION: High TGFbeta2 concentrations in the eyes without intraocular inflammation suggest its immunosuppressive role in human eyes. High concentration of TGFbeta2 (>1,500 pg/mL) was associated with graft acceptance. Also, absence of proinflamatory TNFbeta increased the graft acceptance, but independently from TGFbeta2.

Adult↗

PubMed1998

Water soluble antioxidants in mammalian aqueous humor: interaction with UV B and hydrogen peroxide.

HPLC/electrochemical detection was used to identify five major low MW water soluble electrochemically active molecules from the aqueous humor of three species of mammals: New Zealand White rabbits and humans (diurnal) and Sprague-Dawley rats (nocturnal). These molecules are L-cysteine (CYS), L-ascorbic acid (AA), glutathione (GSH), uric acid (UA) and L-tyrosine (TYR); all of these molecules have known antioxidant properties. Nocturnal rat aqueous humor is concentrated in two thiols: GSH (125 microM; n = 24 pooled eyes) and CYS (63 microM), in contradistinction to diurnal species which have high concentrations of AA. No deterioration of any of these antioxidants occurs in a synthetic aqueous humor mixture irradiated with a physiologically relevant spectral UV B dose of 30 mJ/cm2/h (5.5 UV equivalent sunlight hours). The same result occurred with addition of the endogenous aqueous humor UV B photosensitizer L-tryptophan. In a second set of experiments, human synthetic aqueous humor was subjected to hydrogen peroxide induced oxidant stress. The decay of antioxidants was CYS > GSH > AA > UA > TYR. The second highest concentrated antioxidant in human aqueous humor is TYR. Yet TYR failed to protect AA against H2O2-induced free radical damage in a synthetic aqueous humor model system (P = 0.10; ANOVA). The existence of multiple electrochemically active constituents and their thermodynamic interactions must be recognized when choosing animal models to evaluate human aqueous humor antioxidant defense.

Aged↗

PubMed1981

Aqueous humor lactic dehydrogenase activity in normal and diseased eyes.

Aqueous humor lactic dehydrogenase (LDH) activity has been determined in 13 normal persons and 116 patients with various ophthalmic diseases. Ten postmortem measurements gave abnormally high values and were not used as a control. Aqueous humor LDH activity in 13 normal eyes ranged from 0 to 22.03 mU/ml, the highest activity being seen in retinoblastoma. This result emphasizes the importance of aqueous humor LDH activity determination in differential diagnosis of retinoblastoma from other ophthalmic disorders.

Aqueous Humor↗

PubMed2005

VIP immunoreactivity in human aqueous humor.

PURPOSE: To demonstrate the presence of vasoactive intestinal pep-tide (VIP)-immunoreactive molecule in the human aqueous humor collected from eyes undergoing either cataract or glaucoma surgeries and to identify them asses of molecules responsible for the VIP immunoreactivity. METHODS: Aqueous humor specimens were collected by parencentesis from nine cataract patients and also from eight patients undergoing glaucoma surgery following the creation of the limbal based conjunctival flap, partial dissection of the scleral flap, and application of mitomycin-C. The aqueous humor specimens were analyzed by radioimmunoassay to determine the level of VIP immunoreactivity. Specimens from 10 other cataract patients were pooled and analyzed for VIP immunoreactivity by Western blot analysis. RESULTS: Levels of VIP immunoreactivity in aqueous humor of cataract and glaucoma patients were significantly different and were 610 +/- 160 and 260 +/- 64 pg VIP/ml, respectively (p = 0.03), while there was no correlation between the donor age and the level of VIP immunoreactivity. VIP immunoreactivity was detected as a single molecule with a molecular weight of 9000. CONCLUSIONS: The disease status and the treatments of the eye that led to surgery and procedures applied to the eye immediately before aqueous humor collection, but not the age of the patients, affected the level of VIP immunoreactivity in the aqueous humor. The relationship between the 9000 Da VIP-immunoreactive molecule and the authentic (3326 Da) VIP remains to be studied.

Adult↗

PubMed2001

Aqueous humor flow in normal human eyes treated with brimonidine and timolol, alone and in combination.

OBJECTIVE: To compare the effect on aqueous humor flow and intraocular pressure (IOP) of topically applied 0.2% brimonidine tartrate with topically administered 0.5% timolol maleate, alone and in combination. DESIGN: A randomized, double-masked, placebo-controlled study of 20 human subjects was carried out. The topical drugs were instilled twice daily the day before and again on the morning of the day of the measurements. Aqueous humor flow was measured by clearance of topically applied fluorescein with a fluorophotometer, and IOP was measured with an applanation tonometer. RESULTS: Brimonidine reduced the aqueous humor flow by 33.1%; timolol, by 49.9%; and the combination of brimonidine and timolol, by 58.9%. Brimonidine reduced the IOP by 20.3%; timolol, by 22.9%; and the combination of brimonidine and timolol, by 34.7%. CONCLUSIONS: Brimonidine suppressed aqueous humor flow, but not as effectively as timolol. However, the effects on the IOP of both drugs separately were comparable. The short-term effect of brimonidine was partly additive to timolol, and the combination treatment caused a further reduction in aqueous humor flow and IOP. The IOP reduction by timolol could be explained solely by aqueous humor flow reduction. Much of the IOP reduction caused by brimonidine, but not all, could be explained by suppression of the aqueous humor flow, suggesting an additional mechanism for the ocular hypotensive effect of brimonidine.

Administration, Topical↗

PubMed2005

Determination of glucose in human aqueous humor using Raman spectroscopy and designed-solution calibration.

Glucose concentrations of in vitro human aqueous humor (HAH) samples from cataract patients were determined using 785 nm Raman spectra and partial least squares (PLS) calibration. PLS models were created from spectra of prepared calibration solutions rather than aqueous humor samples. Spectra were obtained with an excitation energy (100 mW for 150 s), which was higher than can be applied in vivo, to decrease the models' contribution to prediction uncertainty. The solutions contained experimentally designed levels of glucose, bicarbonate, lactate, urea, and ascorbate. Multiplicative signal correction of spectra helped compensate for the +/-20% drift in laser power observed at the sample over six noncontiguous days of data collection. Seventeen HAH samples containing 38-775 mg/dL of glucose exhibited a root-mean-square error (RMSEP) of 22 mg/dL, coefficient of determination (r(2)) of 0.989, and bias of 6 mg/dL when predicted from lower energy (30 s) spectra collected contemporaneously with fifty calibration spectra. Similar results were obtained even when spectral data were gathered separately for human aqueous humor samples and calibration samples: 10 HAH samples, calibrated on 25 solutions measured 3.6 weeks earlier, exhibited an RMSEP of 23 mg/dL, r(2) of 0.992, and bias of 9 mg/dL. The results demonstrate progress toward the determination of glucose levels in patient-derived aqueous humor using laboratory-derived "artificial aqueous humor" calibration solutions.

Aqueous Humor↗

PubMed1987

The effects of forskolin on cyclic AMP, intraocular pressure and aqueous humor formation in rabbits.

Forskolin was used to study cyclic AMP-mediated regulation of aqueous humor dynamics in rabbits. Crystalline forskolin was solubilized in oil and its pharmacological effects were studied both in vitro and following topical ocular administration. In vitro, using cultured corneal epithelial cells, forskolin rapidly stimulated cyclic AMP production and in vivo increased cyclic AMP concentration in the aqueous humor 10-fold following topical administration. The effect of topical forskolin on intraocular pressure and aqueous humor formation was determined in vivo using pneumatonometry and fluorophotometry, respectively. Forskolin caused a prolonged reduction of intraocular pressure and decreased aqueous humor formation. The ability of forskolin to potentiate the ocular hypotensive effect of epinephrine was investigated. Forskolin in combination with epinephrine caused a decrease in intraocular pressure of longer duration than either 0.1% epinephrine or 1% forskolin administered separately. Forskolin caused a small but significant increase in the permeability of the blood-aqueous barrier at the time of maximal intraocular pressure reduction. This effect on the blood-aqueous barrier may explain the inhibitory effect of forskolin on aqueous humor formation.

Animals↗

PubMed1981

The effect of age on aqueous humor formation in man.

The rate of aqueous formation was measured with fluorophotometry in a group of 113 normal human subjects who ranged in age from 20 to 83 years. A slight decrease in the rate of aqueous humor formation with aging was detected, but the rate was remarkably stable. The rate of aqueous humor formation was more stable than intraocular pressure or anterior chamber volume in this group and more stable than the tonographic C value reported by other investigators.

Adult↗

PubMedDate not supplied

The composition and optical rotary dispersion of bovine aqueous humor.

The concentration of over 20 biochemicals in bovine aqueous humor was determined by conventional means, and the contribution of each to the observed optical rotation was estimated. Of those studied, only glucose and the total proteins made a significant contribution. However, other unidentified chemicals apparently also contribute to the optical activity since it was not possible to reconstruct the actual aqueous humor optical rotation based on the chemicals included. Alternative experimental approaches that may yield more complete results are discussed. With further work, optical rotation might be used as the basis of a noninvasive sensor for measuring the glucose concentration of aqueous humor in the intact eye.

Animals↗

PubMed1998

[Hydrogen peroxide in the aqueous humor: 1992-1997].

Ideas prevailing in 1991 on hydrogen peroxide in the aqueous humor are outlined. They are critically examined under the light of our finding that the method used to establish aqueous humor levels of peroxide generates itself peroxide during the short time span of the analysis. This is due to the fact that the probe used, dichlorophenol indophenol (DCPIP), spontaneously auto-oxidizes in the presence of oxygen. It was concluded then that the level of hydrogen peroxide in the aqueous humor cannot be higher than about 0.3 microM, the detection limit of the DCPIP method. It was also concluded that the statement commonly made in the literature that aqueous humor hydrogen peroxide derives from the oxidation of ascorbate, an abundant component of that fluid, is based solely on the use of the DCPIP method, and so could easily be due to a methodological artifact. The same applies to the statement that the levels of hydrogen peroxide are very high in human senile cataracts. The surprising resistance to accept the results and conclusions of our 1992 publication is documented. Finally, the content is discussed of an oral presentation made at the 1997 ARVO Annual Meeting in which an important portion of our results and conclusions was confirmed, perhaps signaling a shift towards a wider acceptance of our findings.

2,6-Dichloroindophenol↗

PubMed1999

Population pharmacokinetics of 2% topical dorzolamide in the aqueous humor of humans.

PURPOSE: To evaluate the concentration and kinetics of dorzolamide in the aqueous humor after its topical application. METHODS: Samples of aqueous humor were collected at the beginning of routine cataract surgery at defined intervals after topical application of a 2% solution of dorzolamide. After deep-frozen storage of the samples, drug extraction was achieved with a mixture of solvents. Quantification was carried out by high-performance liquid chromatography on a reversed-phase column. RESULTS: Peak concentrations of dorzolamide in aqueous humor were reached approximately 2 hours after application with 1000 ng/ml. Average values were approximately 1000 to 700 ng/ml after 4 to 6 hours and approximately 200 ng/ml after 12 hours. Mean half-life of absorption was 1.2 hours and for elimination 3.0 hours. CONCLUSIONS: Pharmacokinetics of dorzolamide in the aqueous humor of humans are in comparable dimensions as previously reported in experimental trials in pigmented rabbits. There is a clear linear absorption and elimination kinetic, which is demonstrated using the Bateman function. A better knowledge of the distribution and kinetics of dorzolamide will help to explain its reported effects on intraocular hemodynamics, distinct from its intraocular pressure lowering effect.

Absorption↗

PubMed1998

Endothelin-1 effects on aqueous humor dynamics in monkeys.

PURPOSE: To report data showing no effect of endothelin-1 on aqueous humor formation and total outflow facility in the cynomolgus monkey, in contrast to the enhancement of outflow facility in monkeys and the suppression of aqueous humor flow in rabbits previously reported by others. METHODS: Living monkeys received, unilaterally: (i) intracameral endothelin-1 (10 microl bolus, or 2ml or 4ml exchange; final intracameral concentration 1 or 10 nM) with total outflow facility measured for up to 1 hr post-treatment by two-level constant pressure perfusion, and (ii) intravitreal endothelin-1 (20 microl, final intravitreal concentration 0.1 or 1 microM) with aqueous humor flow measured by scanning fluorophotometry for 5 hr starting 12 hr post-treatment. Contralateral eyes received vehicle. RESULTS: Endothelin-1 had no effect on total outflow facility or aqueous humor flow with any of the methods described. CONCLUSION: Endothelin-1 seems have a variable effect on aqueous humor dynamics within and between species.

Animals↗

PubMed1994

Retinoic acid transport to lens epithelium in human aqueous humor.

Retinoids, in particular retinoic acid, are required for the normal growth and maintenance of many types of cells, including epithelial cells. The metabolism of lens epithelial cells, which are exposed to aqueous humor in the anterior chamber, is thought to be dependent upon aqueous humor dynamics. In human eyes undergoing cataract surgery, we measured retinoic acid levels using reverse-phase high performance liquid chromatography to investigate the possible role of aqueous humor in the transport of retinoic acid to lens epithelial cells. The retinoic acid level in the aqueous humor and the lens epithelial cells was 23.3 +/- 2.3 pmol/ml and 1.8 +/- 0.8 pmol/micrograms protein, respectively. Fluorescence spectra study suggested the presence of endogenous retinoid-protein complexes in the aqueous humor. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the major protein in aqueous humor was albumin, a natural carrier protein of retinoic acid in the blood. From these results, we conclude that the aqueous humor may supply retinoic acid to the lens epithelial cells in human eyes.

Aged↗

PubMed2001

Diabetic cataract and the total antioxidant status in aqueous humor.

Some mechanisms have been proposed for cataract formation in diabetes mellitus such as excessive tissue sorbitol concentrations, abnormal glycosylation of lens proteins and increased free radical production in the intraocular region. We measured total antioxidant status and uric acid levels in aqueous humor from diabetic (n=20) and non-diabetic subjects (n=16) with cataracts. The patients with diabetes and cataract had significantly lower aqueous humor total antioxidant status than those with senile cataract (p = 0.001). Serum and aqueous humor uric acid levels were significantly lower in the diabetic cataract group compared to the senile cataract group. In the diabetic cataract group, the aqueous humor antioxidant status correlated positively with the aqueous humor uric acid levels (p < 0.05). The results of this study suggest that reduced aqueous humor antioxidant status might be associated with reduced aqueous humor uric acid in patients with diabetic cataract. This decrease in aqueous humor uric acid levels might lead to the acceleration of cataract formation.

Aged↗

PubMed2004

Elevated homocysteine levels in aqueous humor of patients with pseudoexfoliation glaucoma.

PURPOSE: To determine total homocysteine levels in aqueous humor of pseudoexfoliation open-angle glaucoma patients. DESIGN: Case-control study. METHODS: Total homocysteine levels were measured by enzyme-linked immunosorbent assay in aqueous humor and plasma of 29 patients with pseudoexfoliation glaucoma and 31 control patients with cataract. Patients with factors affecting homocysteine levels were excluded. RESULTS: We observed significantly elevated (twofold) homocysteine levels in the aqueous humor of the glaucoma patients (Z = -5.11, P <.0001). Additionally, the calculated ratio (plasma:aqueous humor) was significantly lower in these patients (Z = -3.57, P <.001), and aqueous homocysteine was significantly correlated with their respective elevated plasma levels (r =.42, P =.02). CONCLUSIONS: Because homocysteine induces vascular injury and alterations of extracellular matrix, high aqueous homocysteine may trigger the abnormal matrix accumulation characteristic. It may reflect the proposed impairment of the blood-aqueous barrier of pseudoexfoliation open-angle glaucoma and be involved in its pathogenesis.

Aged↗

PubMed2003

Effect of ibopamine on aqueous humor production in normotensive humans.

PURPOSE: Ibopamine is a prodrug of epinine (deoxyepinephrine) that exhibits activity at dopaminergic and adrenergic receptors. Topical ocular application has been shown to cause mydriasis without cycloplegia and to increase the rate of aqueous humor flow in normotensive human eyes. Mydriasis can interfere with the measurement of aqueous flow. In this study ibopamine's effect on aqueous humor production was measured while making allowance for the potential artifact caused by its mydriatic effect. METHODS: The effects of topical ibopamine on pupillary diameter, aqueous humor flow measured by fluorophotometry, and intraocular pressure were studied in 24 healthy, blue-eyed humans. Ibopamine was administered with and without the alpha-adrenergic antagonist dapiprazole, and its effects were compared with those of tropicamide, with and without dapiprazole in a double-masked, randomized, crossover design. RESULTS: Ibopamine dilated the pupil from a diameter of 3.7 +/- 0.64 (mean +/- SD, n=24) to 7.7 +/- 0.70 mm. Ibopamine, during its peak mydriasis, was associated with a very large increase in the rate of clearance of topically applied fluorescein from the cornea and anterior chamber, an effect that was not associated with tropicamide during its peak mydriasis. The mydriatic effect of ibopamine was completely blocked by dapiprazole, and the increase in fluorescein clearance was partially blocked. When mydriasis was blocked, ibopamine increased fluorescein clearance by 13% (P<0.0001), which was interpreted as an increased rate of aqueous humor production. Compared with placebo and with the tropicamide control, ibopamine decreased intraocular pressure, despite its stimulation of aqueous humor flow. CONCLUSIONS: Ibopamine is in a specific class of drug, along with pilocarpine, epinephrine, and bimatoprost that in humans increases the rate of aqueous humor flow as measured by fluorophotometry. This effect is partly responsible for its ability to increase intraocular pressure in persons suspected to have abnormally low aqueous humor outflow facility. The transient apparent doubling of aqueous humor flow, measured by fluorescein clearance after administration of ibopamine is an artifact of increased fluorescein clearance through the dilated pupil while accommodation is active.

Adrenergic alpha-Antagonists↗