Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “FLUORESCEINS”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 55 records · Page 3Linked to original sources

Plasma fluorescein and fluorescein glucuronide in patients with selected eye diseases.

Systemically administered fluorescein (F) is rapidly transformed to the fluorescent metabolite fluorescein glucuronide (FG). Little is known about how diseases can influence the synthesis or disposition of FG. We studied F and FG in the plasma ultrafiltrate of 75 people who were normal or had diabetes, retinitis pigmentosa, or idiopathic rhegmatogenous retinal detachment. F and FG were determined by high-performance liquid chromatography. The concentration of FG was comparable to F 1 h after an intravenous injection of F, both in normal subjects and in patients with retinitis pigmentosa, which suggests that FG may not be an important contributor to the vitreous fluorescence at that time. At later times FG substantially exceeded F. The concentration of FG was significantly higher in diabetics than in the other groups 14 h after an oral dose of F. Accordingly, the possible effect of disease on plasma dye concentrations should be considered in studies measuring F by fluorescence hours after systemic F administration, since this could influence the intraocular fluorescence irrespective of any alteration in ocular function.

Adult↗

Oral fluorescein angiography with the scanning laser ophthalmoscope in diabetic retinopathy: a case controlled comparison with intravenous fluorescein angiography.

AIM: To compare the performance of oral fundus fluorescein angiography with a confocal scanning laser ophthalmoscope (SLO) with intravenous fundus fluorescein angiography (IVFFA) with a fundus camera in the assessment of sight-threatening diabetic retinopathy. PATIENTS AND METHODS: A total of 25 patients undergoing IVFFA to investigate their diabetic retinopathy were recruited. Participants returned 1 week later and an oral angiogram with the SLO was performed. Six facets of the oral and intravenous angiograms were scored and compared: visualization of the foveal avascular zone (FAZ); branch retinal identification; macular leakage; identification of microaneurysms in areas of macular leakage; peripheral nonperfusion, and leakage from neovascular complexes. RESULTS: Compared to IVFFA, the FAZ was unreliably visualized with oral angiography (Kappa 0.1, 95% CI 0-0.3). In contrast, macular leakage (Kappa 0.78, 95%, CI 0.72-0.83), identification of microaneurysms in areas of macular leakage (Kappa 0.78, 95%, CI 0.72-0.83), and neovascular complexes (Kappa 1.0) were reliably seen. Analysis of the visualization of peripheral nonperfusion was complicated by the finding that profuse dye leakage from neovascular complexes obscured the view of the peripheral retina. If the five angiograms in which this occurred were excluded, oral angiography identified 23 of the 24 eyes in which significant nonperfusion was found on IVFFA. CONCLUSION: Oral angiography with the SLO can provide high-quality angiograms that allow judgments to be made about the presence of treatable diabetic maculopathy, proliferative diabetic retinopathy, and peripheral nonperfusion. In the presence of coexisting macular oedema, it proved to be an unreliable technique with which to investigate foveal ischaemia.

Administration, Oral↗

Fluorescein and fluorescein glucuronide in vitreous: fluorescence and binding properties in vitro.

The total and ultrafiltrable fluorescence were measured after disodium fluorescein and fluorescein monoglucuronide were added to aliquots of human and porcine vitreous, and to aliquots of phosphate buffer with and without 0.2% sodium hyaluronate. No significant binding or quenching of F or FG by vitreous or sodium hyaluronate could be found, indicating that vitreous binding is not a source of error in vitreous fluorometry.

Animals↗

Barrier properties of rat subcutaneous nerves to sodium fluorescein and fluorescein-labelled dextrans of various molecular weights.

Na-fluorescein and fluorescein-labelled dextrans of various molecular weights were injected i.v. to localize the blood-peripheral nerve barrier in rat subcutaneous nerves. Samples were taken from the subcutaneous fascia and skin 5 and 30 min after the injection. In the specimens taken 5 min after the injection, the fluorescence had diffused into the dermal tissue. The intraneural, epineural, and perineural blood vessels contained fluorescent material. The entry of fluorescent material from subcutaneous tissue into the nerves was prevented by the perineurium. The distribution of the tracer was uniform throughout molecular weight range tested (376-150,000). Both dermal and intravascular fluorescence were clearly weaker in the specimens taken 30 min following the injection, particularly if a tracer with a MW lower than 40,000 was used. After bilateral ligature of the renal blood vessels the fluorescence remained more intense, even after 30 min, and it was independent of the molecular diameter of the tracer injected. The location of the fluorescence was not changed by the operation and the intraneural capillaries remained intact. The results indicate that the intraneural capillaries are impermeable to the fluorescent tracers tested. In contrast, all the marker substances passed freely through cutaneous capillaries. The perineurium prevented the diffusion of fluorescent tracers from the surrounding tissue into the nerve.

Animals↗

The normal fundus fluorescein angiogram. IV. A classification of the normal fundus fluorescein angiogram based on angiographic findings in clinically healthy subjects and in diabetics without ophthalmoscopically abnormalities.

A comparison of fundus elements in the posterior pole of the eye in a group of 48 clinically healthy subjects and in a group of 17 insulin dependent diabetics showed a significant overrepresentation (P less than 0.05) of hyperfluorescent and hypofluorescent elements among the diabetics, as estimated from serial fundus fluorescein angiograms at 3 examinations during one year. In the diabetics a significant intraindividual variation of hyperfluorescent elements (6/17 eyes) occurred during one year in contrast to the clinically healthy subjects (0/48)-(P less than 0.01). The normal fundus fluorescein angiogram (i.e. in clinically healthy subjects) is characterized by: a wide physiologic variation of the number of hyperfluorescent elements i.e. 0-4 elements or 0-11 elements as assessed with and without a one tailed 10% percentile as an upper limit. Hypofluorescent elements in variable number may occur in 21% (upper limit 10% percentile) or 31% (without an upper limit) of normal eyes. No intra-individual variation of these hyper- and hypofluorescent elements developed during the observation period.

Adult↗

The normal fundus fluorescein angiogram. V. Intraobserver-, interobserver- and interocular variation of the fundus fluorescein angiogram.

An independent assessment of red free fundus photographs and serial fundus fluorescein angiograms from 11 clinically healthy subjects and 5 diabetics was performed by 3 ophthalmologists twice with an interval of one month and showed no significant interobserver or intraobserver variation (P greater than 0.10). Among 8 clinically healthy subjects, fundus fluorescein angiograms of both eyes disclosed a difference in number and distribution of the angiographic elements (P less than 0.05) between the two eyes in 6 subjects.

Adult↗

Iris fluorescein angiography and fluorescein gonioscopic photography after implantation of Choyce Mk IX anterior chamber lenses.

Patients underwent intracapsular cataract extraction and implantation of a Choyce Mk IX anterior chamber intraocular lens. At fluorescein angiography (FAG) at a mean of 8 months post-operatively, 9 showed leaking from the iridal vessels, and 3 were normal: Three cases were excluded because of factors affecting the iris FAG. At a mean of 37 months 5 still had a slight degree of leakage. No primarily negative iris FAG changed to positive later. Fluorescein gonioscopic photography (FGP) revealed leakage in 29.5% of the tips of the foot plates at a mean of 37 months. No correlation was observable between FGP and gonioscopy. The pupillary deformation quotient DQ (greatest: smallest pupillary diameter) was significantly greater at a mean of 8 months in the eyes with leakage in iris FAG than in eyes without leakage (P = 0.0145). Individually a change both in direction against normalisation and in elongation was recorded during the observation time.

Aged↗

A comparative study of proxymetacaine-fluorescein and lignocaine-fluorescein use during applanation tonometry.

AIMS: To evaluate the relative merits of proxymetacaine-fluorescein (PROX-FLU) and lignocaine-fluorescein (LIG-FLU) when used to perform applanation tonometry. METHODS: This prospective, masked, double blind study assessed several aspects of the tonometry process-the duration of the stinging sensation and degree of discomfort, the extent of reflex lacrimation induced, the need for subsequent tear film manipulation to ensure an accurate tonometry reading, and total time to complete tonometry-for each preparation. RESULTS: PROX-FLU caused significantly less discomfort and reflex lacrimation than LIG-FLU and accurate tonometry was more rapidly completed when it was used. PROX-FLU was preferred by 98% of the study patients. CONCLUSION: PROX-FLU is a well tolerated and useful alternative to the more widely used LIG-FLU mixture.

Adult↗

Quantitative inhibition ELISA for diagnosis of schistosomiasis using human IgG labeled with fluorescein and anti-fluorescein/peroxidase.

Usually mouse monoclonal antibodies are used in inhibition assays for antibody determination. Interference may occur in these inhibition assays due to presence of naturally occurring anti-mouse antibodies in some human serum samples. To avoid such interference, human IgG isolated from a pool of serum samples of S. mansoni patients and highly positive for IgG against S. mansoni soluble egg antigen (SEA) was used in inhibition ELISA for diagnosis of S. mansoni infection. The assay was based on inhibition of binding of human IgG labeled with fluorescein to S. mansoni SEA coating microtitration plates by tested serum samples. Plates were washed and labeled human IgG reacted with SEA was linked to peroxidase enzyme by incubation with anti-fluorescein/peroxidase conjugate. The assay showed 90% sensitivity and 96.3% specificity. The level of inhibition in ELISA showed highly significant positive correlation with stool egg output (Kandall's tau b = 0.512, P < 0.001). To make the assay quantitative, serial dilutions of the highly positive human serum pool, used for preparation of human IgG, were applied in each plate and concentration of anti-SEA antibodies in serum samples tested was calculated from a 4-parameters logistic curve equation. The highly positive serum pool used as a standard was considered to contain one million arbitrary units of immunoglobulins against S. mansoni SEA. Human IgG is expected to be more practical in inhibition assays than mouse monoclonal antibodies due to elimination of interference caused by naturally occurring human anti-mouse antibodies. Also, large amount of human IgG could be purfied from remnants of serum samples highly positive for the proposed antibodies. A higher specificity and sensitivity could be obtained if IgG is isolated by affinity purification instead of ammonium sulphate precipitation. In conclusion, human IgG isolated from highly positive serum samples could be used in sensitive and specific diagnostic antibody determination inhibition assays for diagnosis of infectious and autoimmune diseases.

Animals↗

Relative binding properties of fluorescein and 9-hydroxyphenylfluoron (HPF) with murine monoclonal anti-fluorescein antibodies.

Contribution of the fluorescein (F1) carboxyl group to hapten binding by idiotypically related murine monoclonal anti-F1 antibodies 4-4-20, 9-40 and 12-40 was studied by comparing relative liganded active site properties with bound Fl or 9-hydroxyphenylfluoron (HPF). Kinetic studies revealed similar association rate constants between Fl and HPF to 4-4-20 (approximately 1.1 x 10(7) M-1 s-1); however, the 4-4-20 dissociation rate for Fl was approximately 200 times slower, relative to HPF, which resulted in relative intrinsic affinity values of 1.2 x 10(10) and 6.5 x 10(7) M-1, respectively. Mabs 9-40 and 12-40 also displayed a reduced affinity for HPF and affinity constants of 5.5 x 10(5) M-1 and 6.7 x 10(5) M-1 were obtained from a competitive ELISA. Additionally, previous studies revealed that upon binding Fl, Mabs 4-4-20 (92.1%), 9-40 (44.7%) and 12-40 (73.4%) quenched Fl fluorescence. Similar analyses with HPF resulted in 64.4% and 2.0% fluorescence quenching by 4-4-20 and 12-40, respectively; however, 9-40 increased HPF fluorescence by approximately 24%. Steady-state fluorescence polarization experiments revealed that in solution, Fl (P = 0.019) and HPF (P = 0.048) were polarized to different degrees. When bound, however, Fl and HPF expressed similar polarization values (P approximately 0.455), except 9-40 bound HPF which was significantly depolarized (P = 0.428). Fluorescence lifetime experiments revealed Fl to possess two discrete lifetimes: a 3.96 ns component (free Fl) and either a 0.52 ns (4-4-20), 2.23 ns (9-40) or 0.96 ns (12-40) short component that corresponded to bound Fl. HPF, however, when bound by 4-4-20 or 9-40, was best fit by three discrete exponentials: a relatively long 4.0 ns component, a 1.11 ns lifetime (free HPF) and either a 0.52 ns (4-4-20) or 2.23 ns (9-40) component. Finally, HPF bound by Mab 12-40 exhibited a single lorenzian distributed lifetime of 1.36 ns (+/- 0.43 ns). Results are discussed in terms of Mab active site structure and conformational state dynamics.

Animals↗

Pressure-induced dissociation of fluorescein from the anti-fluorescein single-chain antibody 4-4-20.

Hydrostatic pressure was used to dissociate fluorescein (Fl) from the high-affinity anti-Fl single-chain antibody 4-4-20 (SCA 4-4-20). Fl dissociation was monitored by measuring (1) the shift in the Fl absorption peak, (2) the recovery in Fl fluorescence intensity, which is quenched upon SCA binding, or (3) the decrease in Fl fluorescence polarization. Pressure effects were studied at two different Fl:SCA 4-4-20 molar ratios: 1:1, at which Fl fluorescence quenching was ca. 35% at atmospheric pressure, and 1:5, at which quenching reached 95-97% under the same conditions. In both cases, pressure-induced dissociation was favored by concomitant dilution of protein and ligand. Dissociation constants (KD) at each pressure were calculated on the basis of measurements of Fl fluorescence polarization under pressure. The dependence of KD, and consequently of delta G of dissociation, on pressure permitted calculation of the magnitude of the standard volume change (delta V) involved in the dissociation process. According to this study, delta V of dissociation for the Fl-SCA complex is -50 mL/mol, which corresponds to a 10-times higher value than that found for dissociation of Fl from the intact IgG mAb 4-4-20 [Herron, J. N., Kranz, D. M., Jameson, D. M., & Voss, E. W., Jr. (1986) Biochemistry 25, 4602-4609]. This difference is explained in terms of a higher overall flexibility of unliganded SCA and of a less stable binding site in SCA relative to mAb.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies↗

Comparative evaluation of oral fluorescein angiography using the confocal scanning laser ophthalmoscope and digital fundus camera with intravenous fluorescein angiography using the digital fundus camera.

BACKGROUND: To study the efficacy of oral fluorescein angiography (FA) with confocal scanning laser ophthalmoscope (CSLO). METHODS: Ten patients each of choroidal neovascular membrane, central serous retinopathy, diabetic retinopathy and five normal subjects were evaluated with group 1 - intravenous FA using digital fundus camera (DFC); group 2 - oral FA using DFC; and group 3 - oral FA using CSLO. RESULTS: Third-order branch vessels were identified in 85.7% eyes in group 3 versus 51.4% in group 2 (P = 0.004), image quality was comparable to intravenous FA in 77.1% in group 3 versus 48.5% in group 2 (P = 0.02) and margin delineation was better in 80% eyes in group 3 versus 35% in group 2 (P = 0.01). The foveal avascular zone was clear and intact in 54.2% eyes in group 3 versus 37.1% in group 2 (P = 0.14) and 57.1% in group 1 (P = 0.8). CONCLUSION: Oral FA using CSLO is superior to oral FA using DFC and comparable to intravenous FA using DFC in terms of image quality, branch retinal vessel identification and margin delineation.

Administration, Oral↗

[Prospects of using fluorescein angiography with oral administration of fluorescein in children].

Search for the subjective methods for the diagnosis of diseases of the fundus oculi in children necessitates the development of new methods. The authors are investigating the possibility of fluorescent angiography with oral administration of fluorescein with due consideration for the allergologic history and other conditions that may be contraindications against intravenous administration of the agent to children. Fluorescent angiography was carried out in 15 children aged 4 to 15. The procedure was well tolerated, no complications were recorded.

Administration, Oral↗

High affinity binding of fluorescein isothiocyanate to eosinophils detected by laser scanning cytometry: a potential source of error in analysis of blood samples utilizing fluorescein-conjugated reagents in flow cytometry.

BACKGROUND: In samples of peripheral blood cells processed using the commercial kits for detection of apoptosis based on DNA strand break labeling, a subpopulation of cells characterized by high green fluorescence, similar in intensity to that of apoptotic cells but more uniform, was consistently observed by flow cytometry. The labeled cells had no other features of apoptosis. The labeling was observed regardless of the fixative used and was evident in control samples lacking terminal deoxynucleotidyltransferase. Common to all the kits that generated this labeling pattern was the presence of fluorescein (f) conjugated reagents, f-dUTP, f-avidin, or f-antibody. METHODS: Laser scanning cytometry was used to identify the labeled cells and study the mechanism of labeling. Because it was suspected that the traces of unconjugated f-isothiocyanate (FITC) that may contaminate the reagents were responsible for the labeling, FITC binding affinity to white blood cells was studied. Gel electrophoresis was used to detect the presence of unconjugated FITC in the reagents. RESULTS: After staining with Giemsa, the strongly fluorescent objects were identified as eosinophils with normal morphology and no evidence of apoptosis. The fluorescence was localized exclusively within the cytoplasmic granules. Labeling of eosinophils was observed at 2 nM concentration of FITC, which was over three orders of magnitude lower than that needed to label neutrophils, monocytes, or lymphocytes. Gel electrophoresis of the f-conjugated reagents revealed only minor contamination with FITC. CONCLUSIONS: (1) Trace amounts of unconjugated FITC contaminating the reagents are adequate to strongly label eosinophils thereby introducing experimental bias in analysis of apoptosis and in other studies on blood cells utilizing f-labeled antibodies, e.g., in detecting cytokines. (2) FITC at concentration 2-500 nM can be used as a marker of eosinophiles; (3) Because of high affinity to FITC, eosinophiles (or the protein from these cells) may serve as a means of removing traces of unconjugated FITC from the reagents during their manufacture or prior to use.

Apoptosis↗

A kinetic study on the enzymatic hydrolysis of fluorescein diacetate and fluorescein-di-beta-D-galactopyranoside.

The kinetics of the hydrolysis of fluoresceindiacetate and fluorescein-di-beta-D-galactopyranoside were investigated by thin-layer chromatography. The time course of the concentrations of substrate, monosubstituted intermediate, and product was simulated numerically. The mathematical model takes into account the competition of substrate and intermediate and the accumulation of the intermediate at the enzyme.

Chromatography, Thin Layer↗

Use of fluorescein hydrazide and fluorescein thiosemicarbazide reagents for the fluorometric determination of protein carbonyl groups and for the detection of oxidized protein on polyacrylamide gels.

Highly fluorescent thiosemicarbazide and hydrazide prepared by reaction of fluorescein isothiocyanate with hydrazine or adipic acid dihydrazide have been used to monitor the presence of carbonyl groups in oxidatively modified proteins. After oxidation, proteins react with these reagents under anaerobic conditions in the dark to yield fluorescent protein conjugates (presumably thiosemicarbazones or hydrazones) which can be visualized as fluorescent bands following electrophoresis (0-4 degrees C) on lithium dodecyl sulfate-polyacrylamide gels. These reagents do not react with unoxidized proteins. The conjugates formed dissociate readily at room temperature but are fairly stable at pH 6-9, 0 degrees C. Current data suggest that these reagents will be useful in the detection and quantitation of oxidatively modified proteins in biological systems.

Electrophoresis, Polyacrylamide Gel↗

Probing cellular protein targets of H2O2 with fluorescein-conjugated iodoacetamide and antibodies to fluorescein.

Recent studies suggest that H2O2, at subtoxic concentrations generated in response to the activation of a variety of cell surface receptors, functions as an intracellular messenger. However, the intracellular targets of H2O2 action have not been identified. A procedure to detect proteins with reactive cysteine residues susceptible to oxidation by intracellularly generated H2O2 is now described. This approach is based on the labeling of proteinaceous cysteine with 5-iodoacetamidofluorescein at pH 5.5 and immunoblot analysis of the labeled proteins with antibodies specific to fluorescein. With this procedure, many proteins in human A431 cells were shown to contain reactive cysteines and to be readily oxidized by H2O2 generated in response to cellular stimulation with epidermal growth factor. One of these H2O2-sensitive proteins was identified as protein tyrosine phosphatase 1B.

Antibodies↗