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Dye extravasation and the nature of background fluorescence in sodium fluorescein angiography.

PURPOSE: To obtain better insight into the process of dye extravasation in the human choroid during sodium fluorescein angiography of the ocular fundus. METHODS: Sodium fluorescein angiography using a scanning laser ophthalmoscope was performed on the right eye of five healthy volunteers. The dose of sodium fluorescein was chosen to avoid quenching in the ocular vessels. The time-intensity curve of background fluorescence was measured and its shape compared with that of the time-intensity curve of a retinal artery, representing an intravascular compartment. Significant dye extravasation is expected to cause a marked change in shape of the time-intensity curve of background fluorescence as compared with the intravascular time-intensity curve due to long washout of dye from the interstitial space of the choroid. RESULTS: In all five subjects, the time-intensity curve of background fluorescence and the retinal artery proved to be quite similar in shape until several seconds after peak intensity of the first bolus passage. CONCLUSION: Background fluorescence originates mainly from intravascular compartments during the early phase of the sodium fluorescein angiogram.

Adult↗

Indocyanine green versus fluorescein angiography in the differential diagnosis of arteritic and nonarteritic anterior ischemic optic neuropathy.

PURPOSE: We evaluated in a prospective study the usefulness of indocyanine green (ICG) versus fluorescein angiography in the differential diagnosis between arteritic and nonarteritic anterior ischemic optic neuropathy (AION). METHODS: Simultaneous ICG and fluorescein angiography was performed on 22 eyes with AION. Appearance of both dyes in the choroid and in the retina, laminar flow, venous filling, and complete filling of the choroid were measured independently. Massive delayed choroidal filling corresponding to occluded posterior ciliary arteries was especially assessed, as it is almost always diagnostic of arteritic AION. We considered the choroidal filling as massive delayed if it was still incomplete after the venous filling. RESULTS: We diagnosed 5 arteritic AIONs, confirmed by biopsy, and 17 nonarteritic AIONs. In both types of angiography, 3 of 5 patients with arteritic AION showed massive delayed complete choroidal filling times (44.2, 45.8, and 70 seconds), and patients with nonarteritic AION had normal complete choroidal filling times. Dye appearance at the different angiographic times was similar for fluorescein and ICG angiography (P = 0.95-0.96). CONCLUSION: Fluorescein angiography alone is sufficient to reveal massive delayed choroidal filling time in arteritic AION. For our purpose, ICG angiography provides no additional information.

Aged↗

Photoallergic reaction to fluorescein.

A photoallergic reaction to fluorescein was demonstrated in a male diabetic by photopatch and photoprovocation tests. Sensitization was acquired by tonometries using a fluorescein-containing local anesthetic, preceding fluorescein angiography for diagnosis of diabetic retinopathy. Histopathological examination of skin lesions induced in both photopatch and photoprovocation tests confirmed the diagnosis of photoallergic reaction to fluorescein.

Biopsy↗

Hyperventilation enhances transcapillary diffusion of sodium fluorescein.

Voluntary hyperventilation (HV) provokes hemoconcentration due to a loss of fluid from the intravascular space. In 10 healthy male volunteers the hypothesis was tested whether HV increases transcapillary fluid shift into the interstitial compartment. For this purpose, fluorescent light intensity (FLI) alterations after intravenous injection of sodium fluorescein (Na fluorescein) before and during 3 min of HV were determined. Concomitantly, temperature and microvascular skin flux (laser Doppler fluxmetry, LDF) were recorded continuously. Hematocrit and serum proteins, as markers of hemoconcentration, increased significantly from 41.2 +/- 2.3 to 42.7 +/- 2.0% (p = 0.0023) and from 69.5 +/- 3.4 to 72.9 +/- 3.0 g/l (p = 0.0005, respectively). Skin temperature and LDF showed no changes during HV compared to baseline levels. Interstitial FLI indicating transcapillary diffusion of Na fluorescein was significantly higher (p < 0.001) during HV compared to the values recorded during the baseline period. The exact mechanism of enhanced transcapillary diffusion of Na fluorescein is not known. The distinct increase in FLI without a significant change in microvascular skin flux suggests an HV-induced increase in capillary pressure or an enhancement in capillary permeability for water and small solutes.

Adult↗

An autopsy case of fatal anaphylactic shock following fluorescein angiography: a case report.

We present a rare autopsy case of fatal anaphylactic shock following fluorescein angiography. A 71-year-old Japanese woman undergoing retinal angiography to evaluate diabetic retinopathy died immediately after an injection of sodium fluorescein. Forensic autopsy and post-mortem biochemical analyses revealed an elevated serum level of tryptase which, in the absence of morphologic changes suggesting injury or disease, confirmed the diagnosis of fatal anaphylactic shock. Although serious adverse effects are rare after fluorescein angiography, patients should be observed, with appropriate resuscitation equipment available, for several hours after the administration of fluorescein.

Aged↗

Spectral shift of fluorescein and carboxyfluorescein in the anterior chamber of the rabbit eye following systemic administration.

Systemically administered fluorescein is converted to fluorescein glucuronide. The conjugate is fluorescent and interferes with the measurement of fluorescein in the anterior chamber. Carboxyfluorescein is a hydrophilic derivative of fluorescein. In the rabbit, carboxyfluorescein is not converted as readily to a fluorescent metabolite. Thus, carboxyfluorescein has potential advantages as a quantitative fluorophore for studies of aqueous humor dynamics.

Animals↗

Oral fluorescein angiography in patients with choroidal neovascularization and macular degeneration.

OBJECTIVE: To evaluate image quality of oral fluorescein angiography in patients with age-related macular degeneration and choroidal neovascularization. MATERIALS AND METHODS: Twenty four patients with proven choroidal neovascularization were examined in a prospective observational case series. Confocal oral and intravenous fluorescein angiography were performed using the Heidelberg Retina Angiograph. Oral and intravenous image quality were compared in a masked fashion. RESULTS: Choroidal neovascularization could be diagnosed in all patients using intravenous angiograms. Oral fluorescein angiography accurately detected the presence or absence of choroidal neovascularization in all patients and could determine the classification of choroidal neovascularization, location, and extent and borders in 20 (83.3%) of 24 patients. Early, mid, and late phases of oral angiograms occurred at 8, 15, and 26 minutes, respectively. CONCLUSION: Oral fluorescein angiography using the confocal Heidelberg Retina Angiograph allowed detection of choroidal neovascularization in all patients in this study. Visualization of extent and type of choroidal neovascularization was possible in most eyes with choroidal neovascularization. These findings suggest oral angiography is an excellent screening tool for choroidal neovascularization and allows guidance of treatment in the majority of cases.

Administration, Oral↗

[The adverse reactions and treatment of fundus fluorescein angiography].

PURPOSE: To investigate the safety of fluorescein angiography and to evaluate the examination. METHODS: We retrospectively analyzed 8 344 patients who underwent fluorescein angiography in our hospital in the past four years. The rate of adverse events was calculated and the emergency management for severe cases was discussed. RESULTS: There were 693 cases (8.3%) of adverse events occurring in the patients. Many of them were mild or moderate ones, which were 664 cases in number and were 7.87% in rate. The other 29 cases (0.35%) were severe ones. No special treatment was needed for the mild or moderate adverse events, the symptoms can be relaxed by deep breathing or having a rest for a moment. The symptoms of the severe adverse events can be relieved through anti-hypersuscepibility treatments and no sequelae left. No case of death occurred. CONCLUSION: The adverse events could occur in fluorescein angiography examination. It suggests that we should pay attention to it and be aware of the system conditions of the patients. It is also important for us to grasp the indications and contraindications and to prepare for the emergent events. In summary, fluorescein angiography is generally a safe method for the examination of fundus disease.

Adolescent↗

Vulvar Paget disease: fluorescein-aided visualization of margins.

Two women with extramammary Paget disease of the vulva were given fluorescein intravenously in order to visualize disease margins with the use of an ultraviolet light. As a diagnostic modality, the technique was found to have a positive predictive value of 97.4% and a negative predictive value of 99.9%. The accuracy of the fluorescein technique was compared with gross visualization using a histologic mapping technique. A total of 2424 fields were mapped and the predictive accuracy of each test was determined. A chi 2 test demonstrated fluorescein visualization to be a significantly better predictor of a disease state. Fluorescein visualization is suggested as an adjunct to surgical management of patients with primary vulvar Paget disease.

Female↗

Barriers to fluorescein and protein movement.

Vitreous fluorophotometry assesses the leakage of fluorescein through the primary blood-retinal barrier composed of retinal capillaries and the pigment epithelium. However, other ocular barriers exist that may differentiate between the small molecules of fluorescein and the larger ones of protein. We injected solutions containing carboxyfluorescein and several sizes of fluoresceinated dextran into the subretinal space of rabbit eyes. Carboxyfluorescein diffused rapidly through the retina into the vitreous, but dextrans of a size comparable to proteins crossed the retina very poorly. Large dextrans persisted for days in the subretinal space, although the fluid injected with them disappeared within a few hours. Vitreous fluorophotometry must be interpreted with the knowledge that the retina is much more of a barrier to protein than to fluorescein.

Animals↗

Fluorometric quantification of low-dose fluorescein delivery to predict amputation site healing.

This retrospective study evaluated quantification of skin fluorescein delivery by fiberoptic fluorometry as a means of predicting the healing potential of an amputation site. Fluorometry uses a dual-channel fiberoptic light guide--one channel transmits blue light to excite the fluorescein in the skin under study, and the other transmits emitted fluorescence from the skin to a photomultiplier tube where it is measured. Ten minutes after intravenous administration of sodium fluorescein (4 to 8 mg/kg), fluorometric readings were obtained at more than 100 reading sites. In the 86 cases without preoperative cellulitis at the site of amputation, preoperative fluorometry clearly distinguished between healing and nonhealing sites. Healing sites averaged 79% of the fluorescence of a healthy reference area (dye fluorescence index [DFI] = 79), while failing sites averaged only 27% (p less than 0.01 by ANOVA). In all but one case where the DFI was greater than 42, the amputation healed. In all cases where the DFI was less than 38, the amputation failed. In general, uncertainty was limited to sites with values between these limits. The technique maintained its high accuracy in patients with diabetes and for distal amputations. However, it was not accurate at sites of active cellulitis (12 cases). There were no significant adverse effects from the slow injection of the low dose of fluorescein used for this technique. We conclude that fluorometry is an effective means of predicting healing in patients undergoing amputation.

Amputation, Surgical↗

The amino acid sequence of a fluorescein-labeled peptide from the active site of (Na,K)-ATPase.

(Na,K)-ATPase in an active-transport protein that couples the energy obtained from the hydrolysis of ATP to the transport of Na+ and K+ across animal cell membranes. In order to investigate the enzymatic mechanism of this activity, a peptide derived from the ATP-binding site of (Na,K)-ATPase has been purified and its amino acid sequence has been determined. The peptide was identified by the covalent incorporation of a fluorescent probe, fluorescein 5'-isothiocyanate, into the active site before trypsin digestion of the protein. The labeling of (Na,K)-ATPase by fluorescein 5'-isothiocyanate was associated with the irreversible inhibition of enzymatic activity, and both the labeling of the tryptic peptide and inhibition of activity were prevented when the reaction was performed in the presence of ATP. An apparent KD of 5.7 microM was calculated when the reaction between (Na,K)-ATPase and fluorescein 5'-isothiocyanate was performed under pseudo first-order conditions. The amino acid sequence of the active-site peptide, His-Leu-Leu-Val-Met-Lys-Gly-Ala-Pro-Glu-Arg, is similar to the sequence of a fluorescein-labeled peptide derived from the active site of the sarcoplasmic reticulum Ca2+-transport ATPase (Mitchinson, C., Wilderspin, A. F., Trinaman, B. J., and Green, N. M. (1982) FEBS Lett. 146, 87-92).

Adenosine Triphosphate↗

Conformational changes of renal sodium plus potassium ion-transport adenosine triphosphatase labeled with fluorescein.

We studied conformational changes of purified renal sodium plus potassium ion-transport adenosine triphosphatase (ATP phosphohydrolase, EC 3.6.1.3) labeled with fluorescein isothiocyanate. Fluorescein covalently binds to the alpha-subunit of the enzyme and inhibits the ATPase but not the p-nitrophenylphosphatase activity. Four unphosphorylated and three phosphorylated conformations were distinguished by the level of fluorescence and by the rate of its change (relative fluorescence is shown in percentages). Fluorescence of the ligand-free form (E1, 100%) was increased by Na+ (E1.Na form, 103%) and quenched by K+ (E2.K, 78%) at a site of high affinity (K0.5 for K+ = 0.07 mM). Mg2+ did not alter fluorescence of E1 or E1.Na but raised that of E2.K (E2.K.Mg form, 85-90%). Addition of excess Na+ to the E2.K.Mg form restored high fluorescence but the rate of transition from E2.K.Mg to E1.Na became progressively slower with increasing Mg2+ concentration. Two phosphorylated conformations, (E2-P).Mg (82%) and (E2-P).Mg.K (82%) were differentiated by a faster turnover of the latter form. A third conformation, (E2-P).Mg.ouabain, had the lowest fluorescence (56%) and its formation allowed the binding of ouabain to the phosphoenzyme. Reversible blocking of sulfhydryl groups with thimerosal inhibited the formation of E2.K and (E2-P).Mg.ouabain but not that of the other conformations of the fluorescein-enzyme. The thimerosal-treated fluorescein-enzyme retained K+-p-nitrophenylphosphatase activity, inhibition of this activity by ouabain and ouabain binding. The unphosphorylated enzyme had low (K0.5 = 1.2 mM) and the phosphoenzyme had high affinity (K0.5 = 0.03 - 0.09 mM) for Mg2+ in the absence of nucleotides. Since low and high affinity for Mg2+ alternates as the enzyme turns over, Mg2+ may be bound and released sequentially during the catalytic cycle.

Animals↗

Stimulation by aminooxyacetate of fluorescein uptake in rat renal tubules in vitro: role of intracellular alpha-ketoglutarate.

Uphill base-line uptake of a weak organic anion, fluorescein, in rat renal proximal tubules is stimulated by a transaminase inhibitor, aminooxyacetate (AOA, 1 mM). Considering that an inhibition of cytoplasmic aspartate aminotransferase can be accompanied by an elevation in the cytoplasmic alpha-ketoglutarate (KG) level, it was hypothesized that the effect of AOA on the fluorescein uptake was mediated through an augmentation of the KG gradient across the basolateral membrane, which was maintained owing to the Li-inhibitable KG reuptake in the cells. In order to test this hypothesis, an influence of Li on the stimulatory effects of AOA and KG on the fluorescein uptake was investigated. The hypothesis was supported by results showing that LiCl (5 mM) completely abolished the stimulatory effect of KG and attenuated almost 3 times that of AOA, whereas Li did not affect the base-line fluorescein uptake. Moreover, the stimulatory effects of AOA and KG on the uptake were of the same degree but failed to be additive, suggesting that they share a common mechanism. Modulatory influences of metabolic inhibitors, such as fluoroacetate, malonate, phenylpyruvate and D-malate, as well as of L-lysine on the effects of AOA and KG also were investigated to outline metabolic processes involved in these effects. It is concluded that KG metabolism related to its participation in the malate-aspartate shuttle, but not to its oxidation in the tricarboxylic acid cycle, is of importance for the AOA effect.

Aminooxyacetic Acid↗

A fluorescein and indocyanine green angiographic study of choriocapillaris in age-related macular disease.

OBJECTIVE: To examine the phenomenon of a prolonged choroidal filling phase (PCFP) as seen on fluorescein and indocyanine green (ICG) angiography in patients with early age-related macular disease (AMD). METHODS: One hundred eyes of consecutive patients with early AMD were studied. Patchy and slow choroidal filling in early fluorescein and distinct areas of reduced choroidal fluorescence in ICG angiography were interpreted as PCFP. In addition, associated drusen characteristics and the AMD status of the fellow eye were recorded. RESULTS: A PCFP was observed in 26% of eyes using fluorescein and 32% of eyes using ICG angiography, with good concordance between findings using both techniques (K = 0.9). A PCFP was associated with confluent drusen (P = .01), the presence of focal retinal pigment epithelial-atrophic patches in the study eye (P=.005), and geographic atrophy in the fellow eye (P=.03). Other drusen characteristics and the distribution of visual acuity (P = .90) were not different between eyes with and without PCFP. CONCLUSIONS: A PCFP on fluorescein and ICG angiography is a common feature in early AMD. This sign has been interpreted as indicating reduced choroidal perfusion caused by change in diffusional characteristics of Bruch membrane. A PCFP is a clinical marker for diffuse deposits in Bruch membrane and a risk factor for the development of geographic atrophy.

Aged↗

Correlation of the Schirmer 1 and fluorescein clearance tests with the severity of corneal epithelial and eyelid disease.

OBJECTIVE: To evaluate the correlations of the fluorescein clearance test (FCT) and the Schirmer 1 test with the severity of corneal epithelial and eyelid disease in normal patients and patients with tear film disorders due to meibomian gland disease (MGD) and/or aqueous tear deficiency (ATD). METHODS: Nineteen normal control subjects, 16 patients with MGD associated with rosacea, 21 patients with noninflammatory atrophic MGD, and 43 patients with ATD were enrolled. There was a similar age and sex distribution in each group. Each patient completed a symptom severity questionnaire that consisted of 11 questions and then underwent a panel of diagnostic tests in the following order: assessment of corneal and conjunctiva sensation with the Cochet-Bonnet esthesiometer, FCT, assessment of corneal fluorescein staining, Schirmer 1 test (5 minutes without anesthesia), and biomicroscopic examination of the eyelid margins and meibomian glands. The FCT was performed with a fluorophotometer by measuring the fluorescein concentration in minimally stimulated tear samples collected from the inferior tear meniscus. By studying the best area under the receiver operating characteristic curves, we developed a formula that combined the FCT and Schirmer test results, which we termed the FCT corrected by Schirmer test. RESULTS: The FCT showed stronger correlation with ocular irritation symptoms (r = 0. 35, P <.001), corneal fluorescein staining (r = 0.54, P<.001), and meibomian gland and eyelid pathologic signs than the Schirmer 1 test. A correction factor that was based on the best area under the receiver operating characteristic curves, added to the FCT score, improved its correlation with ocular irritation symptoms, eyelid margin and meibomian gland pathologic signs, and ocular surface sensitivity scores. CONCLUSIONS: Corneal epithelial disease is correlated with decreased aqueous tear production and delayed tear clearance, whereas eyelid and MGD are correlated with delayed tear clearance. The FCT corrected by Schirmer 1 test improves the correlations of the FCT with ocular irritation symptoms, corneal epithelial and eyelid pathologic signs, and corneal and conjunctival sensitivity for patients with MGD and ATD.

Aged↗

Fluorescein binding to normal human serum proteins demonstrated by equilibrium dialysis.

The binding of fluorescein to normal human serum proteins in a physiologic solvent at 37 degree C was measured by equilibrium dialysis. Human serum contained 3.28 X 10(-3)M concentration fluorescein-binding sites, with an average association constant at 37 degree C of 0.54 X 10(4)M(-1). The percentage of total fluorescein bound by human serum proteins ranged from 83% to 53% when the total fluorescein concentration ranged from 6.9 X 10(-5) to 6.2 X 10(-3)M.

Binding Sites↗

Acidosis alters fluorescein permeability. Differential tracer penetration through pigment epithelium.

The permeability of the blood-retinal barrier to infusion of fluorescent tracers was examined in tissue sections of frozen eyes from conscious rats that had breathed either air (control) or 25% carbon dioxide in air for one hour. For all animals the blood-retinal barrier remained impermeable to either carboxyfluorescein or Evans blue, indicating a functionally intact tight junctional barrier during hypercapnia. However, in hypercapnic animals, fluorescein penetrated into the neural retina, mainly via the pigment epithelium. Fluorescein also passed through the pigment epithelium in rats with metabolic acidosis induced by intravenous infusion of ammonium chloride, which lowered arterial blood pH without raising the Paco2 or BP. The results indicate an increased permeability of fluorescein through the cell membranes of the pigment epithelium during respiratory and metabolic acidosis. The effect on fluorescein may be due to a pH-dependent increase in the plasma concentration of the associated, more lipid-soluble form of this weak acid.

Acidosis↗