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The source of proteins in the aqueous humor of the normal rabbit.

Aqueous fluorophotometric, tracer localization and modeling methods were combined to document the existence of a pathway in the normal rabbit for the diffusion of proteins from the ciliary and iridial process stromas through the iris stroma into the aqueous humor of the anterior chamber. A new custom-conjugated tracer, fluoresceinated horseradish peroxidase (F-HRP), was used. Anesthetized rabbits were injected intravenously with F-HRP (250 mg/kg). In some animals, aqueous fluorophotometric and tracer localization studies were performed on the same eyes. Anterior chamber fluorescence was detected 2-10 min post-injection and rose to concentrations of 0.01-0.05 mg/ml 60 min post-injection. Subsequent tracer localization studies of these eyes revealed that the morphologic components of the blood-aqueous barrier were intact, that is, no leakage of F-HRP from the iris vasculature or across the ciliary epithelium was observed. Separate tracer localization studies were performed to examine the time course of the route(s) by which tracer entered the anterior chamber. These studies revealed a "wave" of tracer that migrated from the ciliary and iridial process stromas, through the iris, and arrived at the anterior iris surface approximately 8 min post-injection. A pharmacokinetic model based on the diffusional pathway was developed to describe the time course of the concentration of plasma macromolecules in the ciliary body, iris and anterior chamber. Model predictions were consistent with aqueous fluorophotometric and tracer localization results. The diffusion model can account for a major fraction of protein entering the aqueous humor of normal rabbit eyes.

Animals↗

Ceftizoxime concentrations in human aqueous humor following intravenous administration.

The aqueous humor penetration of the third-generation cephalosporin, ceftizoxime, was determined in 21 patients awaiting cataract extraction, after an intravenous bolus injection of 2 g. The beta-lactam compound achieved a mean peak aqueous humor concentration of 7.8 mg/l two hours after administration with a serum/humor percent ratio ranging between 7 and 16% during the four-hour sampling interval. Therefore, ceftizoxime levels in this extravascular fluid were above the minimum inhibitory concentrations for the majority of sensitive organisms.

Adult↗

[Level of uric acid in aqueous humor of patients with cataract].

Aqueous humor was taken from 32 patients with senile and presenile cataract at the operation by means of anterior chamber puncture. Uric acid was marked by indirect method with uricase. The mean content of uric acid in aqueous humor of patients with cataract was 187.13 mumol/l and in the control group 309.34 mumol/l. The difference between the groups is statistically significant. The results suggest that uric acid as strong endogenous antioxidant may play an important role in pathogenesis of cataract.

Adult↗

[Uric acid level in aqueous humor of patients with cataract].

Aqueous humor was taken from 32 patients with senile and presenile cataract at the operation by means of anterior chamber puncture. Uric acid was marked by indirect method with uricase. The mean content of uric acid in aqueous humor of patients with cataract was 187.13 mumol/l and in the control group 309.34 mumol/l. The difference between the groups is statistically significant. The results suggest that uric acid as strong endogenous antioxidant may play an important role in pathogenesis of cataract.

Adult↗

Effect of rabbit aqueous humor obtained after cataract surgery on collagen gel contraction induced by bovine lens epithelial cells.

We evaluated the effect of specimens of pre- and postoperative aqueous humor on the contraction of collagen gels, and the effect of transforming growth factor-beta(2) (TGF-beta(2)) in postoperative aqueous humor. Rabbit aqueous humor was collected preoperatively and on postoperative day 7. Bovine lens epithelial cells (LECs) were cultured in collagen gel in F-12 nutrient mixture supplemented with 5% fetal bovine serum that contained 10% aqueous humor obtained under various conditions. Gel area was determined on day 4. Gels cultured with the medium that contained phosphate-buffered saline showed a statistically significant contraction after 4 days. Aqueous humor from aphakic or pseudophakic eyes significantly increased contraction, with both specimens having a similar effect. Approximately 60% of the contractile effect of the postoperative aqueous humor was neutralized by anti-TGF-beta(2) antibody. However, the promoting effect of the aqueous humor sampled postoperatively was less than that sampled preoperatively. Although the aqueous humor obtained postoperatively increased the contractility of the LECs, with the level of TGF-beta(2) apparently responsible for much of its effect, the effect was less than that observed in the aqueous humor obtained preoperatively.

Animals↗

Identification of heavy-molecular-weight soluble protein in aqueous humor in human phacolytic glaucoma.

Aqueous humor was obtained by paracentesis at the time of cataract surgery from six patients with phacolytic glaucoma, diagnosed on the basis of acute unilateral open-angle glaucoma associated with an apparently leaking hypermature or mature cataract, and from six control patients with immature cataracts. Three of the latter had primary open-angle glaucoma. Quantities of heavy-molecular-weight (HMW) protein (MW greater than 150 X 10(6)) sufficient to obstruct aqueous outflow were identified in all six phacolytic aqueous humor specimens but in none of the controls. Three of the hypermature cataractous lenses from the cases of phacolytic glaucoma were also examined and were found to have 14-fold greater quantities of HMW protein in their liquefying cortex than were present in the cortex of immature cataractous lenses. These findings, correlated with experimental HMW protein perfusion studies in excised human eyes that we have already reported, strongly suggest that direct obstruction of the aqueous outflow channels by liberated HMW soluble lens protein may be a significant and previously unappreciated factor in the pathogenesis of phacolytic glaucoma.

Aqueous Humor↗

Hemolytic complement activity in aqueous humor.

Hemolytic C1, C4, C3, and C5 and IgG levels were measured in aqueous humor obtained at surgery from human eyes without inflammation or previous surgery (group 1), eyes with previous surgery (group 2), and eyes with anterior uveitis (group 3). Serum hemolytic complement and IgG levels were also compared with aqueous humor levels. Aqueous humor from eyes in group 1 contained functional C1, C4, C3, and C5, but there was relatively little complement in normal aqueous humor when compared with serum. The mean values of all complement components and IgG in aqueous humor increased from groups 1 to 3; aqueous-serum ratios for each complement component and IgG also increased from groups 1 to 3. Eyes in group 3 had the highest complement and IgG levels and in some cases these values approached those found in serum.

Adult↗

Examination of the aqueous humor as a diagnostic aid in anterior uveitis.

The aqueous humor of 20 cats and 17 dogs was examined to determine its importance as a diagnostic aid in anterior uveitis. Aqueous humor cytology and protein concentration differed in acute and chronic anterior uveitis but was not specific enough to be an aid in the diagnosis of feline infectious peritonitis. Cytologic examination of the aqueous humor was of benefit in determining the cause of anterior uveitis in 2 of 7 cases of lymphosarcoma. Cultures of aqueous humor proved negative in uveitis of more than 3 weeks' duration. Pseudomonas sp was cultured from the aqueous humor of a dog with anterior uveitis secondary to a perforating corneal injury.

Animals↗

Aqueous humor dynamics in rats.

BACKGROUND: In order to determine normal outflow facility and aqueous humor production values in Lewis rats, we established methods of measuring aqueous humor dynamics in this animal. METHODS: Outflow facility was determined using anterior chamber infusion with constant pressure. Aqueous humor production was determined by a technique of dilution with FITC-albumin. RESULTS: The mean outflow facility was 0.044 +/- 0.01 microliter/min/mmHg. Rats weighing less than 300 g had lower values than did rats weighing more than 300 g (0.034 +/- 0.006 microliter/min/mmHg vs 0.050 +/- 0.015 microliter/min/mmHg, P = 0.009). The mean aqueous humor production was 0.350 +/- 0.110 microliter/min. The turnover rate of aqueous humor production per minute was 2.23%. CONCLUSIONS: Anterior chamber infusion with constant pressure and the FITC-albumin method allow measurement of the outflow facility and aqueous humor production in Lewis rats. These methods may be useful in assessing aqueous humor dynamics when rats are used as a glaucoma model.

Animals↗

Binding inhibition of angiogenic factors by heparan sulfate proteoglycans in aqueous humor: potential mechanism for maintenance of an avascular environment.

Aqueous humor is a clear fluid, primarily a blood filtrate, which circulates through the anterior chamber of the eye and bathes the cornea. We explored the possibility that components in the aqueous humor play a direct part in maintaining the avascular environment of the cornea. We report here that heparan sulfate proteoglycan (HSPG) was found in bovine aqueous humor and that it directly inhibits binding of basic fibroblast growth factor and vascular endothelial growth factor to cell-surface heparan sulfate. We demonstrate that this holds true for all heparin binding proteins tested but not for epidermal growth factor, which does not bind heparin. Furthermore, we show, with mathematical modeling, that the concentration of HSPG in aqueous humor (approximately 4 microg/ml), when combined with the clearance of aqueous humor from the eye due to circulation, is sufficient to block the binding of heparin binding growth factors to corneal endothelium. This mechanism suggests a physiological process to control bioavailability of angiogenic growth factors in the cornea.

Animals↗

Comparison of methods for estimation of Toxoplasma gondii-specific antibody production in the aqueous humor of cats.

Intraocular production of Toxoplasma gondii-specific antibody in cats has been estimated by comparing the ratio of T gondii-specific antibody in aqueous humor and serum with the ratio of total immunoglobulins in serum and aqueous humor (Goldmann-Witmer coefficient; aqueous antibody coefficient; C value). It has been proposed that in human beings, comparison of the ratio of T gondii-specific antibody in aqueous humor and serum with the ratio of antibodies against a nonocular pathogen in serum and aqueous humor is more accurate than methods using total immunoglobulin quantification. We developed an ELISA for detection of calicivirus-specific antibodies in the serum and aqueous humor of cats. By evaluating calicivirus-specific antibody concentrations in the aqueous humor of healthy and diseased cats, calicivirus was assessed as a nonintraocular pathogen. The ratio of T gondii-specific antibodies in the aqueous humor and serum and the ratio of calicivirus-specific antibodies in serum and aqueous humor were evaluated as a means of estimating intraocular T gondii-specific antibody production. A field strain of feline calicivirus was isolated, cultured, and purified. A calicivirus-specific IgG ELISA was developed for detection of feline calicivirus-specific IgG in serum and aqueous humor. Calicivirus-specific IgG was measured in the serum and aqueous humor from 3 groups of control cats. Results suggested that calicivirus is a nonintraocular pathogen in cats and that calicivirus IgG detected in aqueous humor is attributable to leakage across a damaged blood-ocular barrier.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Fibrinogen concentration in the aqueous humor of buphthalmic rabbits.

Fibrinogen concentration in the aqueous humor of buphthalmic rabbits, AXBU/J, and those of the normal parent strain AX/J are 0.36 +/- 0.14 and 0.08 +/- 0.01 mg/ml, respectively. There was no detectable fibrinogen in the aqueous humor of normal inbred strain III/J rabbits. Among the buphthalmic rabbits, elevated fibrinogen concentration in the aqueous humor was observed in two month old prodromal rabbits, indicating a small leakage of fibrinogen into the anterior chamber before the development of buphthalmia. Protein concentration in the aqueous humor was close to normal, in spite of an elevated level of fibrinogen in the young buphthalmic rabbits. Severe protein leakage was seen in only some of the old buphthalmic rabbits.

Age Factors↗

Aqueous humor dynamics in patients with diabetes mellitus.

PURPOSE: We measured aqueous dynamic variables in subjects with diabetes mellitus and correlated them with severity of retinopathy and metabolic control to determine whether diabetes affects the anterior circulation of the eye as it affects the posterior (retinal) circulation. METHODS: Sixty-one subjects with diabetes mellitus type 1 and 60 subjects with diabetes mellitus type 2 were recruited from the active practice of the Mayo Clinic. Thirty-two normal subjects, divided by age into two overlapping groups of 20 each, served as contemporaneous control subjects. The diabetic subjects were stratified into four groups according to severity of retinopathy. Aqueous humor flow was measured by clearance of topically applied fluorescein with a spectrofluorophotometer; outflow facility was measured by tonography; and intraocular pressure was measured by applanation tonometry. RESULTS: In type 1 diabetics, the mean intraocular pressure was slightly greater (14 +/- 3 mm Hg), compared with control subjects (12 +/- 2 mm Hg [P = .002]), while aqueous humor flow was slightly less (2.5 +/- 0.6 microliter/min), compared with control subjects (2.9 +/- 0.5 microliter/min [P = .023]). In type 2 diabetics, the intraocular pressure was 14 +/- 3 mm Hg, which did not differ from that of control subjects (14 +/- 3 mm Hg [P = .258]). Aqueous humor flow in type 2 diabetics (2.5 +/- 0.7 microliter/min) did not differ significantly from that of the control group (2.5 +/- 0.7 microliter/min [P = .961]). Tonographic facility of outflow was not significantly different in type 1 and type 2 diabetics and the control subjects. There was no significant correlation in aqueous humor flow, intraocular pressure, or tonographic facility of outflow to severity of retinopathy or hemoglobin A1c in either type 1 or type 2 diabetics. CONCLUSIONS: The dynamics of aqueous humor are not affected to any clinically significant extent in the early or middle stages of diabetic retinopathy. However, there is a tendency toward less aqueous humor flow in the advanced stages of retinopathy.

Adult↗

Increased basal levels of free plasminogen activator activity found in human aqueous humor.

PURPOSE: To quantify the basal free t-PA activity in human aqueous humor. METHODS: Human aqueous humor obtained by simplified pipette paracentesis at cataract surgery was tested for free t-PA activity in a revised 15-hour amidolytic assay using a t-PA standard curve (n = 15). Total antigenic levels of PAI-1, the principal PA inhibitor, were determined using an ELISA kit. The available PAI-1 activity was tested indirectly using anti-human PAI-1 antibody blocking before t-PA activity assay (n = 11). Plasminogen activator type was determined by anti-human t-PA and urokinase (u-PA) antibody blocking before activity assay (n = 7). RESULTS: Free PA activity ranged widely (0.072 to 0.47 IU/ml; mean, 0.20 +/- 0.10 IU/ml) and was almost completed inactivated (> or = 89%) by antibody against human t-PA but not by the u-PA antibody. PAI-1 total antigen also ranged widely between (0.25 to 8.0 ng/ml; mean, 2.25 +/- 2.54 ng/ml). However, pretreatment of samples with PAI-1 antibody or by acidification (pH 3.2) to inactivate inhibitors did not increase t-PA activity levels. CONCLUSIONS: A basal-free t-PA activity, which is predominantly t-PA, is present in human aqueous humor at approximately 20 times higher levels than previously found with an earlier assay. This activity is estimated to represent roughly 10% of total released t-PA antigen. PAI-1 is present in aqueous humor at levels considerably lower than reported values for plasma in a predominantly PA-complexed or inactive form.

Aged↗

Concentration of hyaluronic acid in primary open-angle glaucoma aqueous humor.

We compared the concentration of hyaluronic acid in the aqueous humor of primary open-angle glaucoma (POAG) patients and non-glaucomatous patients. Aqueous humor samples were obtained from 22 patients just before trabeculectomy for clinically uncontrolled POAG (POAG group). Aqueous humor (0.1 mL) was aspirated by inserting a 26-gauge needle into the anterior chamber. The same procedure was performed for 22 non-glaucomatous patients just before cataract surgery (control group). Immediately after collection, the aqueous humor was stored at -20 degrees C. The concentration of hyaluronic acid was determined by a sensitive, noncompetitive and nonisotopic fluoroassay. The median (range) concentrations of hyaluronic acid of the POAG and control groups were 298.4 microg L(-1) (99.0-743.7 microg L(-1)) and 545.1 microg L(-1) (145.0-2366.0 microg L(-1)), respectively. The difference in concentrations of hyaluronic acid between the groups was statistically significant (P<0.001). In conclusion, the concentration of hyaluronic acid in the aqueous humor in POAG patients is lower than in non-glaucomatous patients. Further studies are necessary to determine the role of hyaluronic acid in the pathophysiology of POAG.

Adult↗

[Aqueous humor dynamics].

Intraocular pressure is mainly controlled by the rate of aqueous humor production and the resistance to its outflow. Aqueous humor formation is the result of fast unidirectional secretion and slow contradirectional reabsorption. The most striking evidence of endogenous regulation of the ciliary epithelial secretion is provided by the observation of the circadian rhythm. Aqueous humor formation is mainly controlled by the interaction of inhibiting alpha(2)-adrenoreceptors and stimulating beta-adrenoceptors. The role of the central nervous system in the regulation of this process remains unclear and the precise mechanism of outflow regulation is not fully understood. The aqueous humor passes into the anterior chamber and leaves the eye by two routes: the direct outflow pathway through the trabecular meshwork or the indirect outflow pathway through the ciliary muscle. Further biochemical, biological and pharmacological investigations are necessary to determine the fundamental process of aqueous humor formation, outflow and regulation.

Absorption↗

Retinoblastoma aqueous humor: aromatic amino acids.

Aqueous humor from children with retinoblastoma obtained at enucleation and from eyes with adult cataracts were assayed with electrochemical liquid chromatography (Model 5500 Coulochem electrode array system) for metabolites of tyrosine, tryptophan metabolic pathways, catecholamine degradation pathways and ascorbate. More than 20 metabolites were identified in human aqueous for the first time. High levels of ascorbate were found in aqueous of eyes with adult cataracts (254, 336 ng/ml). Tyrosine metabolism in both sets of eyes was through dopamine. Vandylmandelic acid (VMA), homovanillic acid (HVA), and 3-methoxy, 4-hydroxyphenylglycol (MHPG) were all detected in retinoblastoma eyes. Although eyes with either adult cataracts or childhood retinoblastoma convert tryptophan through the serotonin pathway, retinoblastoma eyes metabolize tryptophan through the kynurenine pathway to a greater degree than adult cataract eyes.

Adult↗

Does the aqueous humor have a role in mitogen-activated protein kinase (MAPK) intracellular signaling in Glaucoma?

Glaucoma is a common blinding disease worldwide. Glaucoma treatment today is based on reduction of aqueous humor production or increase aqueous humor drainage. By medical manipulation, the treatment goal is to reduce the main risk factor, elevated intra ocular pressure. Here we hypothesize that the aqueous humor has a role, beside oxygen and nutrient supply, in transferring signaling to the trabecular meshwork. This signaling might be delivered from the ciliary body were the aqueous humor is produced, or from the lens or the cornea. Recently in our lab we proposed that MAPKs present in the aqueous humor are a novel signal involved in glaucoma pathology. Here we show that this pathway exists at an unexpected, extracellular media. Western blot analysis of aqueous humor from congenital glaucomatic rabbits and a rat model of induced elevated intra ocular pressure (IOP) were found to express several signaling members of the MAPK family. Although these members are usually found in an intracellular environment, they can be detected at an extracellular environment, namely the aqueous humor. These signaling proteins are found also in normal eyes. Moreover the MAPK signaling proteins are found in the active phosphorylated form and in non-active form in elevated IOP animals as well as in the control, normal IOP groups. Understanding the signaling cascade, at the aqueous humor, opens a new area for treatment of glaucoma patients. By interfering with the signaling cascade, taking place at a reachable location, the anterior chamber, we will be able to manipulate these protein effects on the trabecular meshwork.

Aqueous Humor↗