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PubMed1995

Fine specificity analysis indicates that the primary and secondary fluorescein-specific cytotoxic T cell receptor repertoires are indistinguishable.

Previous data from this laboratory have shown that fluorescein-specific cytotoxic T cells derived from naive mice are capable of distinguishing between different isomeric forms of the fluorescein hapten, as well as between the iodoacetamido-, isothiocyanate- and dichlorotriazinyl amino-fluorescein derivatives. In this report, we show that T cells derived from previously immunized animals, while demonstrating a stronger and more cross-reactive response overall, display identical fine specificity patterns to those of primary T cells for a panel of fluorescein homologues. We interpret this finding to indicate that the antigen receptor repertoires utilized by primary and secondary cytotoxic T cells are indistinguishable and that the enhanced response of memory as compared with naive CTL results from factors such as a higher density of cell surface receptors or receptor-associated molecules and/or a higher frequency of antigen-responsive T cells.

Animals↗

PubMed1996

The plasma membrane Ca2+ pump mutant lysine591 --> arginine retains some activity, but is still inactivated by fluorescein isothiocyanate.

Inactivation of the wild-type human plasma membrane Ca2+ pump (isoform 4b) by fluorescein isothiocyanate is accompanied by covalent modification of Lys591. The mutation of Lys591 to arginine reduced the Ca2+ transport activity to 35% of the wild-type, and diminished the amount of acylphosphate formed from ATP by a corresponding amount. When this mutant was treated with fluorescein isothiocyanate; the enzyme was still irreversibly inactivated, even though no reactive residue was available at position 591. The results show that, although Ca2+ pump function is sensitive to the residue at position 591, Lys591 is not essential for enzyme activity. They also demonstrate that irreversible inhibition of the plasma membrane Ca2+ pump by fluorescein isothiocyanate does not require the covalent modification of Lys591. This indicates that fluorescein isothiocyanate reacts with lysine residues at other positions in addition to Lys591.

Adenosine Triphosphate↗

PubMed2000

[Simultaneous ICG- and fluorescein-angiography for fundus examination].

BACKGROUND: An indocyanine-green (ICG) angiography is rarely used as the only diagnostic procedure. Almost always it is performed additionally to a fluorescein angiography. The use of simultaneous ICG and fluorescein (SIF) angiography therefore makes sense. Several examples for the application of SIF-angiography are presented. MATERIALS AND METHODS: SIF-angiography was performed using a 2-wavelength scanning laser ophthalmoscope (SLO). Images were digitally recorded in real time with a graphics workstation. The following cases will be presented: choroidal neovascularization in age-related macular degeneration, choroidal hemangioma, inflammatory fundus disease (APMPPE) and idiopathic polypoidal choroidal vasculopathy. RESULTS: ICG and fluorescein angiography can be simultaneously recorded with the 2-wavelenghth SLO. The quality of the combined pictures is comparable to single-channel recordings of separate ICG and fluorescein images. We show results of the above mentioned cases. CONCLUSIONS: SIF angiography is time efficient and allows precise comparison and analysis of the transit of both dyes through retinal and choroidal circulation. The topographic relation of pathologic findings in ICG angiograms with the critical retinal vascular landmarks is facilitated.

Adult↗

PubMed2002

Fluorescein electronic endoscopy: a novel method for detection of early stage gastric cancer not evident to routine endoscopy.

BACKGROUND: Fluorescence endoscopy with fluorescein sodium in the stomach was evaluated by using a newly developed fluorescence electronic endoscopic system. METHODS: Sixteen patients with early stage gastric cancer diagnosed by white light endoscopy and chromoendoscopy underwent fluorescein electronic endoscopy before surgery. The resection specimens underwent thorough histopathologic evaluation. RESULTS: About 10 seconds after intravenous injection of fluorescein, fluorescence appeared and immediately spread throughout the gastric surface. A few minutes later, differentiated early stage gastric cancers with more abundant stroma than surrounding normal mucosa exhibited significantly stronger fluorescence, and those with less stroma exhibited weaker fluorescence than the surrounding normal mucosa. Undifferentiated early stage gastric cancers, in which the stroma became wider because foveolae were collapsed from malignant invasion, expressed stronger fluorescence intensity. In all cases, the borders of early stage gastric cancers were clearly demonstrated. Among the 16 patients, 6 accompanying flat lesions and 1 tiny lesion not evident by routine endoscopy were detected. The extent of the cancers, as determined by fluorescence endoscopy, were similar to those determined histopathologically. CONCLUSIONS: Fluorescein electronic endoscopy is useful in determining the extent within the mucosa of gastric cancers when this is obscure by standard endoscopic observation, and for detecting extremely early stage cancer that is not evident by conventional endoscopic observation.

Contrast Media↗

PubMed2002

Detection of fungal organisms in eosinophilic mucin using a fluorescein-labeled chitin-specific binding protein.

BACKGROUND: The ability to identify fungal hyphae in patients with chronic rhinosinusitis (CRS) has been inconsistent. A new fluorescein-labeled staining method targets chitin found in fungal cell walls. OBJECTIVE: We hypothesize that this method would be able to more consistently detect fungi within the mucin of CRS patients. METHODS: Fifty-four consecutive CRS surgical patients were evaluated. After ensuring sensitivity and specificity of this new method, all specimens were stained with either fluorescein-labeled chitinase or Grocott methanamine silver stain for comparison. RESULTS: All 54 specimens contained eosinophilic mucin on hematoxylin and eosin staining. One or more fungal hyphae could be visualized within the mucin of 54 (100%) of 54 specimens stained using the fluorescein-labeled chitinase. Only 41 (76%) of 54 of the specimens stained with the Grocott methanamine silver stain technique demonstrated fungi. CONCLUSION: The fluorescein-labeled chitinase-staining technique has greater sensitivity in detecting fungal organisms within eosinophilic mucin. Fungal organisms are present in the mucin of CRS patients.

Adolescent↗

PubMed1981

Fluoresceinated chemotactic peptide and high-affinity antifluorescein antibody as a probe of the temporal characteristics of neutrophil stimulation.

Antifluorescein antibody molecules were used to interrupt the stimulation of neutrophils by a fluoresceinated chemotactic peptide. From the results we construct a semiquantitative relationship among ligand-receptor interaction, the time course of cell triggering and response, and aspects of cellular adaptation. The interaction of the antibody with the free fluoresceinated peptide is complete within a few seconds and the peptide-antibody complex neither stimulates the cells nor inhibits subsequent stimulation by unlabeled peptide. When antibody is added to a cell suspension that has been stimulated with the fluoresceinated peptide, we observe that: (i) the apparent membrane depolarization response monitored by a fluorescent dye can be inhibited only if antibody is added within 30 sec of stimulation; (ii) the superoxide response can be inhibited even if antibody is added more than 1 min after stimulation and decays with an intrinsic half-life of about 12 sec; (iii) responses to a second dose of nonfluoresceinated peptide are enhanced if the antibody is added within 2 min of stimulation by the fluoresceinated peptide. These results suggest that different neutrophil responses depend in individual ways on the time course and extent of ligand binding to its receptor. A comparison of these data with the time course of binding permits an estimate of the number of receptors involved in these responses.

Antigen-Antibody Complex↗

PubMedDate not supplied

Synthesis and study of the fluorescein conjugate of the nucleotide dPTP.

The synthesis of a fluorescein conjugate on the non-natural base P is described. The ability of the newly synthesised fluorescein--dPTP and of a fluorescein-11-dUTP to compete with the natural nucleotide TTP was also studied. Overall the efficiency of labelling a nucleic acid with a fluorescein moiety was found to be approximately equal.

Binding, Competitive↗

PubMed1998

Quantifying changes in retinal circulation: the generation of parametric images from fluorescein angiograms.

Fluorescein angiography is an established technique for examining the functional integrity of the retinal circulation. The ability to quantify this function offers the possibility of early detection of changes due to retinopathy. We have developed a technique to generate functional, parametric images of the retinal circulation. A given angiogram is first registered to align consecutive frames. At each point in the retina, a graph of fluorescein intensity versus time is then constructed and fitted with a gamma variate curve. Parameters are extracted from these curves and formed into parametric images showing the variation in fluorescein passage across the entire area of the angiogram. Parameters examined to date include time to maximum intensity, time of arrival and rise time. The technique has been demonstrated using photographic and scanning laser ophthalmoscopic angiograms of both normal subjects and patients with a variety of retinopathies. The time to maximum images of the normal subjects reveals a similar fillings pattern in each case, whilst the pathologies present in the abnormal angiograms are clearly identified. The generation of functional time to maximum images enables the health of the retinal circulation to be quantified with respect to the rate at which the vasculature fills with fluorescein. This offers a potential tool for detecting the onset of retinopathy and monitoring its progression.

Fluorescein Angiography↗

PubMed1995

1.85 A structure of anti-fluorescein 4-4-20 Fab.

The crystal complex of fluorescein bound to the high-affinity anti-fluorescein 4-4-20 Fab (Ka = 10(10) M-1 at 2 degrees C) has been determined at 1.85 A. Isomorphous crystals of two isoelectric forms (pI = 7.5 and 7.9) of the anti-fluorescein 4-4-20 Fab, an IgG2A [Gibson et al. (1988) Proteins: Struct. Funct. Genet., 3, 155-160], have been grown. Both complexes crystallize with one molecule in the asymmetric unit in space group P1, with a = 42.75 A, b = 43.87 A, c = 58.17 A, alpha = 95.15 degrees, beta = 86.85 degrees and gamma = 98.01 degrees. The final structure has an R value of 0.188 at 1.85 A resolution. Interactions between bound fluorescein, the complementarity-determining regions (CDRs) of the Fab and the active-site mutants of the 4-4-20 single-chain Fv will be discussed. Differences were found between the structure reported here and the previously reported 2.7 A 4-4-20 Fab structure [Herron et al. (1989) Proteins: Struct. Funct. Genet., 5, 271-280]. Our structure determination was based on 26,328 unique reflections--four times the amount of data used in the previous report. Differences in the two structures could be explained by differences in interpreting the electron density maps at the various resolutions. The r.m.s. deviations between the variable and constant domains of the two structures were 0.77 and 1.54 A, respectively. Four regions of the light chain and four regions of the heavy chain had r.m.s. backbone deviations of > 4 A. The most significant of these was the conformation of the light chain CDR 1.

Amino Acid Sequence↗

PubMed2002

Intraoperative assessment of perilymphatic fistulas with intrathecal administration of fluorescein.

OBJECTIVES: Assessment of perilymphatic fistulas remains a diagnostic problem. Because of a lack of reliable clinical tests, exploratory tympanotomy is necessary to detect membrane leaks in the middle ear. To improve objectivity in the intraoperative visualization of perilymphatic fistulas, we used intrathecal fluorescein for perilymph staining to increase the sensitivity and specificity of diagnosis of perilymphatic fistulas. STUDY DESIGN: Prospective study in the setting of a tertiary, referral, and academic center. METHODS: Twenty-eight patients with suspected traumatic, idiopathic, iatrogenic, or inflammatory perilymphatic fistulas were admitted for exploratory tympanoendoscopy for perilymphatic fistula detection. Twenty-five to 100 mg sodium fluorescein 10%, diluted with cerebrospinal fluid, was administered by lumbar puncture 2.5 to 23.5 hours before tympanoendoscopy. The oval and round window niches were microscopically and endoscopically observed with white and blue light, using specific filters. RESULTS: In two patients (7%), obvious fluorescence was detected behind the round window membrane with blue light. Direct observation of perilymph in stapedectomy and in semicircular canal fistula revealed no staining. Neurological complications of intrathecal fluorescein, as reported by other authors, were not observed. CONCLUSIONS: Probably because of a different patency of the cochlear aqueduct, intrathecal fluorescein for intraoperative detection of perilymphatic fistula resulted in a significant perilymph staining in only a few patients. Considering the possible complications of this method, we would not recommend it for routine evaluation of perilymphatic fistula.

Adult↗

PubMedDate not supplied

Fluorescein angiography of choroidal malignant melanomas with retinal invasion.

The authors reviewed the fluorescein angiograms of 99 choroidal malignant melanomas to determine the frequency of several angiographic abnormalities and the effectiveness of fluorescein angiography in identifying retinal invasion by the tumor. They identified a "double circulation" pattern in 60 of the 99 cases, broadened intercapillary spaces in 54, large zones of retinal capillary obliteration or obscuration in 35, blockage or obscuration of the larger caliber blood vessels overlying the tumor in 25, and tumororetinal vascular anastomoses in 15. Twelve of the 13 eyes in which they identified fluorescein angiographic abnormalities that they interpreted as evidence of definite or probable retinal invasion and that came to enucleation shortly after the angiography were confirmed to have retinal invasion histopathologically. Awareness of the fluorescein angiographic features indicative of retinal invasion may increase the frequency with which this manifestation of choroidal malignant melanomas is recognized.

Adult↗

PubMed1993

Fatal acute myocardial infarction after intravenous fluorescein angiography.

Since its introduction, fluorescein angiography has been widely used to investigate diseases of the ocular fundus. A case of fatal acute myocardial infarction after intravenous fluorescein angiography is presented. This appears to be the first case documented by autopsy in which the findings are compatible with myocardial infarction as the cause of death. Although there are no known contraindications for fluorescein angiography in patients with a history of cardiovascular disease, the indications for this elective procedure should be carefully reviewed in such patients. Adequate emergency resuscitation equipment should be available in the fluorescein angiography suite.

Aged↗

PubMed1994

Preoperative fluorescein angiographic features of surgically removed idiopathic epiretinal membranes.

PURPOSE: To determine whether preoperative fluorescein angiograms can be used to predict visual prognosis in eyes undergoing surgical removal of idiopathic epiretinal membranes. METHODS: A retrospective study of preoperative fluorescein angiographic findings in cases of idiopathic epimacular membrane removal during vitreous surgery was conducted. Of 422 consecutive cases, 229 met the criteria for minimum follow-up period of 6 months. Angiograms from these 229 cases were analyzed in a standardized, masked fashion for macular edema, retinal vascular distortion, and contraction of the foveal avascular zone. RESULTS: Visual improvement was greatest in 7 eyes (3%) with the most severe degree of macular edema (P = 0.0215). No difference in visual outcome was noted between eyes with less severe macular edema or no macular edema. Change in visual acuity was not associated with the extent of retinal vascular distortion (P = 0.477) or contraction of the foveal avascular zone (P = 0.248). CONCLUSION: The presence of macular edema on preoperative fluorescein angiography is not predictive of a poor visual outcome in eyes undergoing surgery for idiopathic epiretinal membranes. In general, preoperative fluorescein angiography is not useful in predicting visual prognosis in these cases.

Adolescent↗

PubMed2000

Phototoxicity from systemic sodium fluorescein.

PURPOSE: To confirm the occurrence of phototoxicity due to systemic fluorescein in a dose consistent with retinal angiographic testing and to approximate the prevalence of this reaction in a small group of volunteers. METHODS: Fourteen volunteers underwent a controlled challenge test by applying a topical sunscreen and exposing skin areas to direct sunlight before and after fluorescein administration. RESULTS: Two subjects experienced marked cutaneous blanching erythema, mild vesiculation, and pain in sun-exposed areas within 1 hour of exposure. The reaction faded over a period of days without tanning. CONCLUSIONS: Consistent with its in vitro properties as a photodynamic dye, fluorescein may rarely act as a phototoxic agent in humans at doses employed for fluorescein retinal angiography. It is unclear why this reaction has not been reported more frequently, given the large number of patients undergoing this ophthalmic procedure annually.

Administration, Oral↗

PubMed2002

Variability in fluorescein angiography interpretation for photodynamic therapy in age-related macular degeneration.

OBJECTIVES: To investigate the variability in fluorescein angiography interpretation for photodynamic therapy in age-related macular degeneration. METHODS: Eight graders, who included two TAP-certified ophthalmologists, three other retinal specialists, two fellows in vitreoretinal diseases, and a senior fundus photograph grader, evaluated fluorescein angiograms of six patients treated according to the Treatment for ARMD With Verteporfin (TAP) protocol at a single center. Each patient's baseline angiogram was evaluated to determine whether the CNV lesion was predominantly (> or =50%) classic. For each follow-up angiogram, at 3, 6, 12, and 24 months, the grader was required to determine whether fluorescein leakage was present. Six months after the initial gradings, each reader was again presented with the baseline angiogram for each patient and once again asked to determine whether the CNV lesion was predominantly classic without knowledge of the previous grading. All gradings were performed without knowledge of the clinical course. RESULTS: In grading initial visit and follow-up visit angiograms, the overall concordance rates were 81% and 82%, respectively. Concordance rates were not statistically different between the group as a whole when compared with the gradings of the two TAP-certified ophthalmologists. When initial visit angiograms were regraded, an intraobserver variability of 17% was noted. Overall, gradings were discordant with the majority opinion in approximately 19% of decisions. CONCLUSIONS: Considerable variability can be expected in fluorescein angiography interpretation as the results of the TAP investigation are applied to clinical practice.

Choroidal Neovascularization↗

PubMed2004

Unusual reaction to fluorescein dye in patients with inflammatory eye disease.

PURPOSE: Ocular side effects attributable to intravenous fluorescein dye are not well characterized. The purpose of this report was to describe three patients with an unusual ocular reaction after the intravenous administration of fluorescein dye. METHODS: Retrospective review of the clinical and photographic records of three patients. RESULTS: Each patient had some type of preexisting ocular inflammatory disease. Each patient described the subacute onset of a unilateral burning sensation and tearing several minutes after the administration of intravenous fluorescein dye. Findings included a new onset of or a worsening of unilateral conjunctival chemosis and injection in all three patients and yellowish discoloration of the conjunctiva in two patients. In each patient, the noninflamed (fellow) eye did not develop any symptoms or show any visible reaction. The symptoms and findings resolved promptly in the affected eye without specific treatment or effect on vision. CONCLUSIONS: Ocular side effects of intravenous fluorescein dye can include transient symptomatic burning and tearing associated with conjunctival chemosis, injection, and yellowish discoloration. Eyes with active inflammatory diseases may be predisposed to this rare effect through an unclear mechanism.

Adult↗

PubMed2006

Multifocal electroretinography and fluorescein angiography in retinal vein occlusion.

PURPOSE: To interpret the responses of multifocal electroretinography (mfERG) of the central macula in retinal vein occlusion and correlate them with findings of fluorescein angiography, optical coherence tomography (OCT), and visual acuity. METHODS: In a prospective observational case series, 25 patients with branch or hemiretinal vein occlusion were examined by means of mfERG, fluorescein angiography, OCT, and visual acuity. RESULTS: In eyes with macular ischemia shown by fluorescein angiography, the implicit time was significantly longer than that in eyes without macular ischemia (P = 0.006; Mann-Whitney test). The amplitudes in the first-order kernel responses in the ischemic eyes were more reduced (P = 0.04; Mann-Whitney test), and the amplitudes in the central macula correlated well with visual acuity (P = 0.006; Pearson correlation). There was no significant correlation between the degree of retinal thickening as measured by OCT and the mfERG responses, although there was a trend toward reduced amplitudes in the maculae with central retinal thickening (P = 0.08; Pearson correlation). CONCLUSION: Macular ischemia as measured by fluorescein angiography correlates well with the prolonged implicit time on the first-order kernel response in mfERG, but no significant correlation was detected between the responses of mfERG and the OCT findings. Further investigations will demonstrate if this can be used as a prognostic instrument in the selection of patients, who will most likely benefit from future treatments.

Adult↗

PubMed1984

Experimental study on the circulatory dynamics of the spinal cord by serial fluorescein angiography.

The circulatory dynamics on the dorsal surface of the spinal cord were observed by serial fluorescein angiography . Fluorescein dye appeared in the posterior spinal artery and pial arterial plexus, although the direction of blood flow might be opposite and the time taken for fluorescein dye to enter the arteries was different for each. Fluorescein dye appeared in the veins after various periods, and the directions of flow of the dye were also variable and complicated. Laminar flow often was observed in veins of less than 200 microns in diameter. The regional circulation time was calculated to be 2.1 +/- 0.4 seconds. There was no extravasation.

Animals↗