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PubMed1990

Oral fluorescein angiography: reassessment of its relative safety and evaluation of optimum conditions with use of capsules.

Injection of fluorescein intravenously for fundal angiography is associated with a high incidence of minor adverse effects (21%) but a very low incidence of serious (life threatening) reactions (0.05%). A serious reaction may occur without warning in a patient with no history of atopy. There are no reports of oral fluorescein causing a serious reaction, and minor adverse effects are uncommon. A study was undertaken to determine optimum conditions for oral fluorescein angiography. Capsules proved more convenient than a solution for ingestion of fluorescein. A dose of 25 mg/kg body weight produced good quality angiograms in 75% of the patients. The best pictures were obtained by photographing the fundus after 40 and 60 minutes.

Administration, Oral↗
PubMed1983

Platelet-fibrin embolism in a rhesus macaque: angiographic and pathologic studies comparing fluorescein and indocyanine green.

A male rhesus macaque was found to have what appeared to be numerous platelet-fibrin emboli in retinal vessels in the perimacular area. Indocyanine green (ICG) dye fluorescence and fluorescein angiograms of the fundus demonstrated leakage of fluorescein, but not ICG, from the involved arterioles. Histopathologic changes in the eyes included occlusion of retinal and choroidal vessels with platelet-fibrin emboli, inner retinal ischemia, ischemic injury to the parafoveal capillary bed distally to occlusion of precapillary arterioles, and retinal exudate limited to the regions of capillary damage. Differential leakage of fluorescein may be explained by the difference in binding affinities of the 2 dyes to blood protein: 20% to 40% of the circulating fluorescein is unbound, and 98% of ICG is bound to serum albumin. Simultaneously or serially performed angiograms with fluorescent probes of different sizes might be used to obtain a qualitative measure of vascular integrity in persons with embolism, diabetic retinopathy, sickle cell retinopathy, vasculitis, and other disorders known to produce focal retinal and choroidal vascular occlusion.

Animals↗
PubMed1995

The effect of sodium fluorescein on argon green photocoagulation.

BACKGROUND: The absorptive and photosensitizing properties of sodium fluorescein (NaFl) injected prior to laser photocoagulation may affect the retinal laser burn threshold. METHODS: Photocoagulation was performed using an argon green laser with a Volk QuadrAspheric fundus lens in three anesthetized rabbits. Using a 500um spot size and a 100msec duration, varying laser powers were delivered before and at various intervals after intravenous (i.v.) injection of NaFl. RESULTS: Photocoagulation effects were enhanced in all subjects after fluorescein injection with threshold reduced for up to 48 hours following injection. CONCLUSIONS: The effectiveness of photocoagulation following fluorescein angiography may be underestimated, although this may be clinically insignificant since most photocoagulation procedures are performed well above threshold. Nevertheless, these data should be considered in those situations where carefully controlled threshold burns are desired. In such circumstances, it is advisable to take additional care in selecting treatment powers if fluorescein has been injected or to delay photocoagulation for a minimum of 48 hours.

Absorption↗
PubMed1994

Adverse reactions during retinal fluorescein angiography.

BACKGROUND: Fast sequence retinal fluorescein angiography is a commonly employed diagnostic procedure within the optometric practice with relatively few serious adverse reactions. A retrospective study was conducted to document the incidence of adverse reactions with this procedure. METHODS: A total of 1,173 patient charts who had undergone intravenous injection for retinal fluorescein angiography at a specility referral clinic or a referral clinic at a school of optometry. All patients had been intra-venously injected with 500 mg of sodium fluorescein in 25% or 10% solution. Adverse reactions were noted within the charts. RESULTS: The most common adverse reaction were nausea (.8% of patients) and urticaria (.6% of patients), with other reactions including emesis and hypoglycemia. Extravasation of dye was noted in .2% of patients. No acute anaphylaxis was noted. CONCLUSIONS: Fast sequence retinal fluorescein angiography is a relatively safe diagnostic test. However, one should be prepared to handle acute anaphylaxis within the office before administering the test because of previously published cases of life-threatening reactions.

Adolescent↗
PubMed2004

Absorption and paracellular visualization of fluorescein, a hydrosoluble probe, in intact house sparrows (Passer domesticus).

We describe a method to visualize the cellular location of compounds during absorption by the small intestine in intact animals. First, we employed pharmacokinetic methodology to measure the fractional absorption of sodium fluorescein, a small (MW = 376) water-soluble molecule that is widely used as hydrophilic marker molecule for paracellular permeability studies. Based on the hypothesis that the paracellular pathway acts as a sieve, we predicted that fluorescein absorption would be considerable, but less than that of passively absorbed L-glucose which is a smaller molecule (MW = 180). When the two compounds were gavaged into house sparrows simultaneously, the birds absorbed significantly less fluorescein (42 +/- 8%) than L-glucose (82 +/- 7%), as predicted, and absorptions of the two were correlated as one would predict if they shared the same pathway. We removed intestinal tissue 10 min after gavage with sodium fluorescein and determined the cellular location of the compound's fluorescence using confocal laser microscopy. The fluorescent signal was found primarily in the paracellular space. In contrast, in the same type of experiment using instead the similar-sized fluorescent lipophilic compound rhodamine 123 (MW = 381), most fluorescence appeared inside enterocytes, as expected for a compound that diffuses across the apical membrane. Thus, results from all the experiments are consistent with the hypothesis that hydrophilic fluorescein is absorbed primarily via a paracellular pathway. These methods could be applied to visualize absorption pathways of other compounds in other intact animals.

Journal Article↗
PubMed1985

Experimental retinal detachment. X. Effect of acetazolamide on vitreous fluorescein disappearance.

Retinal detachments were created in one eye of each of eight cynomolgus monkeys. Total vitrectomy was performed in the fellow eyes. Fluorophotometry was used to study the rate of disappearance of fluorescein injected into the vitreous cavity. The rate of fluorescein loss via the anterior chamber accounted for only 1% to 3% of the total rate of vitreous fluorescein loss in eyes with retinal detachment or in fellow eyes. Posterior loss of fluorescein (presumably across the retinal pigment epithelium) increased by 25% following intravenous acetazolamide in eyes with retinal detachment and 22% in fellow eyes. It is concluded that acetazolamide increases the rate of fluid absorption across the retinal pigment epithelium.

Acetazolamide↗
PubMed1987

The antiviral effects of rose bengal and fluorescein.

We evaluated the antiviral effects of rose bengal and fluorescein sodium. The direct antiviral activity was determined by an in vitro direct neutralization assay. The 50% inhibitory dose was 16 micrograms/mL for rose bengal and 460 micrograms/mL for fluorescein. The in vivo antiviral effects of these drugs were determined in the mouse herpetic keratitis model. Following topical application, rose bengal reduced surface virus titers (swabs) 1 million-fold, and residual ocular virus (eye homogenates) 32-fold, compared with controls. No infectious virus was recovered by swabbing after topical application of rose bengal. Fluorescein had no significant effect on virus replication. Thus, rose bengal, unlike fluorescein, has significant antiviral activity, and the diagnostic use of rose bengal prior to viral culture may preclude a positive result. Also, the use of rose bengal to grade keratitis in the study of new antiviral agents should be discouraged.

Animals↗
PubMed2004

Ultrabright fluorescein-labeled antibodies near silver metallic surfaces.

Fluorescein-labeled antibodies are widely used in clinical assays and fluorescence microscopy. The fluorescent signal per labeled antibody is limited by fluorescein self-quenching, which occurs when the antibody is heavily labeled with multiple fluoresceins. We examined immunoglobulin G (IgG) when labeled with 0.7 to about 30 fluoresceins per antibody molecule. The extent of self-quenching was decreased, and the signal increased, when the labeled antibody was in close proximity to metallic silver particles. Time-resolved measurements showed that the intensity increase was due in part to a silver-induced increase in the radiative decay rate. These results suggest the use of labeled antibodies conjugated to silver particles as ultrabright probes for imaging or analytical applications.

Antibodies↗
PubMed1989

Three-dimensional structure of a fluorescein-Fab complex crystallized in 2-methyl-2,4-pentanediol.

The crystal structure of a fluorescein-Fab (4-4-20) complex was determined at 2.7 A resolution by molecular replacement methods. The starting model was the refined 2.7 A structure of unliganded Fab from an autoantibody (BV04-01) with specificity for single-stranded DNA. In the 4-4-20 complex fluorescein fits tightly into a relatively deep slot formed by a network of tryptophan and tyrosine side chains. The planar xanthonyl ring of the hapten is accommodated at the bottom of the slot while the phenylcarboxyl group interfaces with solvent. Tyrosine 37 (light chain) and tryptophan 33 (heavy chain) flank the xanthonyl group and tryptophan 101 (light chain) provides the floor of the combining site. Tyrosine 103 (heavy chain) is situated near the phenyl ring of the hapten and tyrosine 102 (heavy chain) forms part of the boundary of the slot. Histidine 31 and arginine 39 of the light chain are located in positions adjacent to the two enolic groups at opposite ends of the xanthonyl ring, and thus account for neutralization of one of two negative charges in the haptenic dianion. Formation of an enol-arginine ion pair in a region of low dielectric constant may account for an incremental increase in affinity of 2-3 orders of magnitude in the 4-4-20 molecule relative to other members of an idiotypic family of monoclonal antifluorescyl antibodies. The phenyl carboxyl group of fluorescein appears to be hydrogen bonded to the phenolic hydroxyl group of tyrosine 37 of the light chain. A molecule of 2-methyl-2,4-pentanediol (MPD), trapped in the interface of the variable domains just below the fluorescein binding site, may be partly responsible for the decrease in affinity for the hapten in MPD.

Antibodies, Monoclonal↗
PubMed1987

Plasma fluorescein decay determination during fluorophotometry.

Two useful methods for determination of the decay curve of non-protein bound fluorescein (NPBF) in plasma up to 1 hour after intravenous fluorescein injection are described and evaluated. The course of NPBF is approximated in method 1 by a sum of two exponential decay functions and in method 2 by a power of time function. The parameters in these functions are calculated with the use of concentration values measured in two blood samples taken at about 5 min. and 60 min. after injection. Calculations in method 1 include the amount of fluorescein injected. The accuracy of each method was evaluated in 7 volunteers by measuring NPBF concentration in 15-28 blood samples taken after fluorescein injection at intervals of 5 min. or less. The mean relative deviation between calculated and measured concentration values amounted to 9.2% +/- 4.3 SD and 12.7% +/- 4.5 SD for method 1 and 2, respectively. The time integral of NPBF concentration in plasma up to one hour after injection was calculated according to the results of both methods and compared with integral values obtained by linear interpolation between concentration values measured in the 15-28 plasma samples. The mean relative deviation for the 7 volunteers amounted at 15 min. to 2.8% and 17% and at 60 min. to 11% and 18% for method 1 and 2, respectively. The maximal difference between the blood-retinal barrier permeability value for NPBF calculated with and without taking glucuronation into account was estimated to be 20% for an average glucuronation percentage of 70% or less.

Adolescent↗
PubMed1989

Effect of antihypertensive treatment on blood-retinal barrier permeability to fluorescein in hypertensive type 1 (insulin-dependent) diabetic patients with background retinopathy.

The effect of antihypertensive treatment on blood-retinal barrier leakage of fluorescein in background retinopathy was studied in nine hypertensive Type 1 (insulin-dependent) diabetic patients suffering from nephropathy. The patients were investigated before and after 7 (3 to 13) months of treatment with captopril (n = 8; 25 to 100 mg daily) and a diuretic, either frusemide (n = 4; 80 to 200 mg daily) or bendrofluazide (n = 2; 2.5 mg daily). Retinal function was assessed by fundus photography, fluorescein angiography, vitreous fluorometry, and renal function by glomerular filtration rate, and albuminuria. The antihypertensive treatment induced a significant reduction (p less than 0.05) in: blood pressure from 152/97 +/- 14/8 mmHg to 134/82 +/- 11/6 mmHg; blood-retinal barrier leakage of fluorescein from 2.4 +/- 1.1 to 1.4 +/- 0.5.10(-7) cm/second; albuminuria from 1391 (range: 168-4852) micrograms/min to 793 (range: 35-2081) micrograms/min. Glomerular filtration rate declined from 88 +/- 15 to 78 +/- 23 ml.min-1.1.73 m2 (0.05 less than p less than 0.10). The metabolic control of the patients as reflected by their blood glucose and HbA1c levels remained stable during the study. Our study suggests that systemic blood pressure elevation contributes to the abnormal blood-retinal barrier permeability to fluorescein characteristically found in diabetic background retinopathy and that this abnormality can be reversed during antihypertensive treatment.

Adult↗
PubMed1985

Serum PABA and fluorescein in the course of Bz-Ty-PABA and pancreolauryl test as an index of exocrine pancreatic insufficiency.

Forty-six subjects (20 chronic pancreatitis, 7 chronic liver disease, 7 recovered from acute pancreatitis, 2 Crohn's disease, and 10 healthy controls) classified by S-C test as having normal pancreatic function (26 subjects), or moderate (10 subjects) and severe (10 cases) pancreatic insufficiency, were given, on different days, 1 g of oral PABA or 348 mg of oral fluorescein dilaurate. At the 1st, 2nd, and 4th hours (PABA) and the 2nd, 4th, and 6th hours (fluorescein) serum samples were taken for assay. In the presence of severe exocrine pancreatic insufficiency, the sensitivity of the fluorescein serum levels was higher than that observed for the PABA (100% and 80%, respectively), and quite similar to that shown by the urinary tests (100% and 70%, respectively). On the contrary, in presence of moderate pancreatic insufficiency, both the urinary test (pancreolauryl and (PABA) give a sensitivity higher than that found in the serum tests (30-40% and 10-30%, respectively). The parallel combination of both the serum or urinary tests does not significantly improve the sensitivity of the single test. These results suggest that the serum PABA and serum fluorescein tests can be valid choice when a prolonged urinary collection is difficult, i.e., in children and in elderly patients. However, the slight diagnostic gain does not justify the routine use of both urinary and serum tests.

4-Aminobenzoic Acid↗
PubMed2005

Fluorescein leakage of the optic disc in glaucomatous optic neuropathy.

PURPOSE: To identify and quantify the role of capillary leakage of the optic nerve head in digital fluorescein angiography in normal subjects and patients with open-angle glaucoma. METHODS: We conducted a prospective cross-sectional study in the Department of Ophthalmology of the Technical University of Aachen. Thirty patients with primary open-angle glaucoma (POAG) and 30 healthy age-matched subjects were included. Fluorescein angiograms were performed using the scanning laser ophthalmoscope. The fluorescence of the optic nerve head and the surrounding retina (ratio of leakage) was measured using digital imaging analysis in the late phases of the angiogram (9-10 min). RESULTS: The ratio of optic nerve head fluorescence to retinal reference loci was significantly increased (p=0.01) in patients with glaucoma (POAG, 1.38+/-0.34) compared with normal subjects (1.20+/-0.19). Intraocular pressure (p=0.0001), visual field indices (mean deviation, p<0.0001; pattern standard deviation, p<0.0001; corrected pattern standard deviation, p<0.0001), and cup to disc ratios (p=0.02) differed significantly between the groups. Age and systolic and diastolic blood pressure showed no significant differences between groups. CONCLUSION: Fluorescein angiography revealed significantly increased vascular leakage of glaucomatous optic nerve heads. An endothelial disruption and fluorescein leakage might be the result of mechanical stress at the level of the lamina cribrosa and/or a sign of ischemic damage. This measurement approach might enable us to judge the severity of optic nerve head leakage, and it is a potential way to evaluate therapeutic regimens.

Blood-Retinal Barrier↗
PubMed1979

Plasma complement and histamine changes after intravenous administration of sodium fluorescein.

Venous blood samples were obtained from 85 patients before and at three, ten, and 30 minutes after intravenous administration of sodium fluorescein. Whole complement hemolytic activity and plasma histamine were measured. The patients were observed for side effects. Clinical adverse reactions occurred in 18 (21%) of the patients. Changes in complement hemolytic activity occurred in all patients, but probably were the result of binding of fluorescein to complement proteins, or on the membrane of sensitized red blood cells used in the assay. Plasma histamine was increased within the first few minutes after infusion of fluorescein and persisted up to ten minutes. Increased histamine was found in 66% of patients with adverse reactions and in only 15% of patients with no reactions. Histamine may be an important mediator of adverse reactions to fluorescein.

Adolescent↗
PubMed1985

A micropreparation of fluorescein conjugates of immunoglobulin G and Fab from serum.

Protein A-Sepharose CL-4B (PAS) was used to isolate rabbit immunoglobulin G from crude anti-herpes simplex virus-1 serum. Papain treatment of the PAS-bound immunoglobulin G released Fab fragments from the solid support, while Fc-containing fragments remained bound to PAS. PAS-immobilized immunoglobulin G was fluoresceinated by reaction with fluorescein isothiocyanate followed by papain cleavage to yield fluorescein-conjugated Fab fragments in solution. These fragments retained activity toward herpes simplex virus-1 infected Vero cells as evaluated by immunofluorescence. This novel procedure represents the fastest and simplest method for preparing Fab or fluoresceinated Fab fragments directly from any volume of immune serum.

Animals↗
PubMed1988

Variations in human corneal endothelial cell morphology and permeability to fluorescein with age.

Fluorophotometry with topically applied fluorescein and endothelial cell photography were performed on 80 normal subjects (age 5-79 yr). Variations in endothelial cell morphology and function, flow of aqueous humor, and intraocular pressure were recorded. The mean endothelial cell size was 332.3 +/- 46.3 micron 2. A 28% increase in endothelial cell size was measured over the eight decades (r = 0.53, P less than 0.001). The coefficient of variation of cell size also increased with age (r = 0.41, P less than 0.001). The percentage of hexagonal endothelial cells decreased by 14% (r = -0.48, P less than 0.001), while the percentage of pentagonal and heptagonal cells increased by 50% (r = 0.44, P less than 0.001) and 40% (r = 0.33, P less than 0.002), respectively, with age. The mean endothelial permeability to fluorescein was 4.03 +/- 0.63 x 10(-4) cm min-1. A 23% increase in endothelial permeability with age was observed (r = 0.44, P less than 0.001). No change in central corneal thickness or endothelial pump rate was found. Flow of aqueous humor remained stable with age, despite a 25% increase in intraocular pressure (r = 0.50, P less than 0.001). Polarization of fluorescence of fluorescein in the corneal stroma decreased with age (r = -0.46, P less than 0.001). We conclude that with age the human corneal endothelium becomes morphologically less regular and may become more permeable to fluorescein.

Adolescent↗
PubMed1986

Assessment of the fluorescein dye test for prediction of skin flap viability in pigs.

The visual fluorescein dye test for prediction of actual skin flap viability was evaluated in pigs. Two delayed random (4 X 10 cm) and arterial (4 X 20 cm) skin flaps were constructed on one flank of a pig, and four mirror-image skin flaps were raised acutely on the other flank of the same pig. Sodium fluorescein dye (15 mg/kg) was injected intravenously 1 and 18 hr after raising of flaps. The maximum length and area of dye stain in these flaps (N = 24) were assessed under Wood's lamp illumination, 15 min after dye injection. The actual maximum lengths and areas of skin survival of these flaps in the same pig were measured 7 days postoperatively. It was observed that visual fluorescein dye test performed 1 hr after surgery significantly (P less than 0.05) underestimated the maximum length and area of actual skin survival. On the other hand, when the fluorescein dye test was performed 18 hr postoperatively, the maximum length of dye stain and the maximum length of actual skin viability were highly correlated (r = 0.97, N = 24, P less than 0.01). Their mean values were similar (10.6 +/- 0.8 vs 10.8 +/- 0.7 cm, mean +/- SEM), and their mean coefficient of variation was 5.6 +/- 1.6%. Similarly, there was a high degree of correlation (r = 0.89, N = 24, P less than 0.01) between the maximum area of dye stain and actual skin flap viability. Their mean values were similar (68.3 +/- 4.5 vs 71.8 +/- 3.9%), and their mean coefficient of variation was 8.7 +/- 2.4%.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1983

Effect of sodium fluorescein on neurite outgrowth from the retinal explant culture: an in vitro model for retinal toxicity.

Effect of sodium fluorescein was examined on neurite outgrowth from the retinal explants of chick embryos cultured with chicken gizzard extract, containing macromolecules which promote neurite outgrowth. A dose more than 0.5 mg/ml of fluorescein completely prevented the neurite outgrowth from the retinal explants. The minimum dose of toxic effect of fluorescein on the neurite outgrowth was about 0.2 mg/ml. The fluorescein also caused lysis of pre-existing neurites which had been fully extended by gizzard extract.

Animals↗