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PubMed1985

Fluorescein angiography in inflammation of the peripheral fundus: involvement of the choroid and retina. II.

Fluorescein angiographies were performed in 65 patients with peripheral uveitis. 34 subjects (52%) showed pathologic dye transit of the peripheral choroid. Fluorographic evidence of retinal involvement was high (91%), preferentially consisting in leakages from capillaries and venules. Fluorescein angiography was able to detect minute lesions and corresponded with the activity of the inflammation.

Choroiditis↗
PubMed2006

Fluorescein-enhanced autofluorescence thoracoscopy in patients with primary spontaneous pneumothorax and normal subjects.

RATIONALE: The exact site(s) and pathophysiology of air leakage in patients with primary spontaneous pneumothorax (PSP) are unknown. In one patient with PSP, fluorescein-enhanced autofluorescence thoracoscopy (FEAT) has shown areas of parenchymal abnormality unnoticed during white light thoracoscopy (WLT). OBJECTIVES: To prospectively perform and compare WLT and FEAT in patients with spontaneous pneumothorax and in normal subjects. METHODS: One-time FEAT and WLT inspection with systematic mapping of semiquantified lesions in 12 consecutive patients with PSP was compared with one-time FEAT and WLT during sequential bilateral thoracoscopy in 17 control subjects. RESULTS: WLT abnormalities (anthracosis, cobblestone malformation, and blebs/bullae) were more prevalent in PSP. FEAT, however, showed high-grade lesions in PSP only, which often were present at areas that were normal, or that only showed anthracosis at WLT. When blebs/bullae were present, bleb-associated FEAT abnormalities were only present in two. Actual fluorescein leakage was seen in two patients with PSP. CONCLUSIONS: Lungs in patients with PSP show significantly more abnormalities at WLT when compared with normal subjects. High-grade FEAT lesions were exclusively present in PSP, and predominantly at lung zones that appeared normal at white light inspection. These findings suggest that significant parenchymal abnormalities are not limited to lesions visible during WLT, such as blebs and bullae.

Adult↗
PubMed2007

Quantitative fluorescein angiographic analysis of choroidal neovascular membranes: validation and correlation with visual function.

PURPOSE: To compare computerized analysis with traditional grading methods in the analysis of fluorescein angiograms from patients with choroidal neovascularization (CNV) due to age-related macular degeneration (AMD) and to examine the clinical relevance of parameters generated by computerized analysis by testing their relationships with clinical measures of vision. METHODS: Custom quantitative fluorescein analysis (QFA) software was used to analyze 62 angiograms from patients with CNV for whom distance visual acuity (DVA) data were available. On applying QFA, we obtained three mathematical parameters for each lesion component: pixel area (PA), integrated intensity (II), and positive fluorescence (PF). Quotients (Q) were derived for the latter two parameters by correcting against background (b) or optic nerve (o) fluorescence (IIQ(b), IIQ degrees , PFQ(b), and PFQ degrees ). The new metrics were compared with traditional grading parameters of classic CNV and lesion area. The relationships of both sets of angiographic data with measures of vision were explored by regression analyses. RESULTS: Weighted kappa between QFA and traditional grading for lesion subtype assessment was high (kappa = 0.7). Regression analyses with PA, IIQ(b), IIQ degrees , PFQ(b), and PFQ degrees for each lesion descriptor (leakage, classic CNV, occult CNV, total lesion) as independent variables and DVA as the dependent variable showed that in every case PFQ(b) exhibited the most significant relationship with vision (adjusted r(2) = 0.26). Parameter estimates showed that for a change of 30 units on the PFQ(b) for classic CNV, a loss of 20 letters of DVA may be expected. No parameters from traditional grading methods showed statistically significant relationships with DVA. CONCLUSIONS: The markers of dynamic change in area and intensity of fluorescence exhibited stronger relationships with visual function than did area measurements alone.

Choroidal Neovascularization↗
PubMed1991

Isoxsuprine in nonproliferative diabetic retinopathy: effect on human retinal circulation assessed by fluorescein angiography--a pilot study.

The retinal vessel filling time of 10 patients with early nonproliferative diabetic retinography was measured by fluorescein angiography. Initially, the reproducibility of the authors' standardized fluorescein angiography was assessed by control angiograms on two different dates in each patient. Thereafter, prolonged use (six months) of isoxsuprine sustained release (80 mg/day) led to a nearly 50% reduction of the arterial filling time and a nearly 25% reduction of the arteriovenous filling time, whereas no changes were seen in control patients. This effect of isoxsuprine might be explained by an increase of red cell deformability and, concomitantly, a decrease of platelet adherence in the microcirculation of patients with diabetic mellitus.

Adult↗
PubMedDate not supplied

An endoscopic method of tympanic membrane fluorescein angiography.

Fluorescein angiography is a recently developed method of studying the tympanic membrane's dynamic vasculature. Our preliminary studies using an ophthalmic fundus camera and aural speculum did not always permit visualization of the entire tympanic membrane, and the electronic flash unit used in our initial studies did not cycle fast enough to visualize effectively the rapid arterial phase of blood flow. In this report, we describe an endoscopic method of tympanic membrane fluorescein angiography that uses a continuous xenon light source. A representative angiogram from a normal subject is described. The mallear artery apparently is the major blood supply to the posterior half of the tympanic membrane, which is consistently better perfused than the anterior half. Branches from the annular ring of blood vessels supply the anterior half of the tympanic membrane. Preliminary studies of two temporalis fascia tympanoplasty patients indicate that the graft becomes revascularized 2 to 4 weeks after surgery and that posterior grafts may revascularize earlier than anterior grafts.

Adult↗
PubMed1997

Detection of phosphatidylinositol glycan class A gene transcripts by RT in situ PCR hybridization. A comparative study using fluorescein, Texas Red, and digoxigenin-11 dUTP for color detection.

Gene-specific probes labeled with fluorescein, Texas Red, and digoxigenin-11 dUTP (DIG) were used for RT in situ PCR hybridization to detect PIG-A gene (phosphatidylinositol glycan class A) transcripts. The PIG-A gene is responsible for biosynthesis of the glycosylphosphatidyl-inositol (GPI) anchor. Lack of GPI anchor expression due to mutations can cause an acquired clonal hematologic disorder called paroxysmal nocturnal hemoglobinuria (PNH). In this RT in situ PCR study, two types of labeling methods, a direct method (using fluorescein and Texas Red) and an indirect method (using DIG-11 dUTP) were compared. Both were successfully applied to detect and localize the PIG-A gene transcripts within single cells of the cell lines AA2, H9, and JY. Furthermore, similar results for sensitivity and reproducibility were obtained. Advantages and disadvantages of the different labeling techniques are discussed. In addition, peripheral blood mononuclear cells from PNH patients were also included in this study.

Cells, Cultured↗
PubMedDate not supplied

Establishment of fluorescein diacetate and ethidium bromide (FDAEB) assay for quality assessment of isolated islets.

One of the most important factors in clinical islet transplantation is isolation of a great number of islets with good viability. According to viability assessments of isolated islets, the static incubation test and the perifusion test of islets, which are used retrospectively, take much time and need various apparatus. But viability assessments of isolated islets for clinical islet transplantation require a simple, rapid, sensitive, and prospective method. We have developed a microfluorometric viability assay for isolated human, porcine, and dog islets of pancreata using fluorescein diacetate and ethidium bromide (FDAEB). Fluorescein diacetate (FDA) causes live cells to fluoresce green under blue light excitation (490 nm) and ethidium bromide (EB) causes dead cells to fluoresce red. In this study, we investigated the applicability of FDAEB staining to quality assessment of isolated islets for clinical use by correlation with the counting method with insulin secretion of islets. Discrimination of living from dead islets by insulin secretion correlated well with viability as determined by FDAEB staining. The proportion of living islets within isolated canine islets, as measured by microfluorometric counting, was found to correlate highly significantly on low-temperature (24 degrees C) culture (R = 0.831, p < 0.001) and on 37 degrees C culture (R = 0.553, p < 0.05) with the insulin contents of the same islets. Therefore, it is possible to differentiate degrees of viability, and a scoring system is described for this purpose. The FDAEB assay prospectively and easily provides a rapid, accurate, and objective measurement of the proportion of living cells and dead cells in isolated islets for clinical islet transplantation.

Animals↗
PubMedDate not supplied

Fluorescein dye disappearance test in patients with different degrees of epiphora.

PURPOSE: To assess the diagnostic efficacy of a modification of the fluorescein dye disappearance test (DDT) in patients with different degrees of epiphora. METHODS: DDT was performed by instilling a drop of 2% fluorescein in the eye. Schirmer tear test strips were placed in the lower conjunctival fornix, after 3 and 10 minutes. The colour dilution on the strips was compared to a scale with known standards and the most closely comparable colour dilution was chosen and recorded as the visual scale test (VST) grade of the strip. The grey-scale value (GV) of each strip was calculated by computer image analysis. The control group comprised 44 patients, mean age 44.8 years (SD 7.5, range 30-60). There were 13 patients in group 1 with moderate epiphora; mean age was 44.2 years (SD 9.5, range 30-60). There were 17 patients in group 2 with severe epiphora; mean age was 48.3 years (SD 7.7, range 37- 60). RESULTS: VST and GV differed significantly between controls and group 1 patients at 3 minutes (p=0.002, p=0.001), and between controls and group 2 patients at 3 minutes (p<0.001, p<0.001). There was no difference between groups 1 and 2 patients in VST and GV at 3 minutes. The differences were highly significant between controls and group 1 at 10 minutes (p=0.001, p=0.007), and between controls and group 2 at 10 minutes (p<0.001, p<0.001). The differences in GV and VST between groups 1 and 2 were significant at 10 minutes (p=0.026, p=0.018). CONCLUSIONS: This modified DDT permits identification of different degrees of epiphora in a non-invasive manner, it can therefore be used to evaluate the outcome of different treatment strategies.

Adult↗
PubMed1979

Polarization of fluorescein fluorescence in single cells.

Measurement of fluorescence polarization (P) gives information about the immediate environment of the fluorescent molecule. We used a flow polarimeter to investigate the factors influencing P of fluorescein in mammalian cells to determine whether such measurements are useful for characterizing heterogeneous cell populations. Fluorescein was introduced into cells by incubation with FDA. Measurements of the intensity of fluorescence (TI) and polarization (P) revealed an unexpected dependence: P decreased with increasing intensity of fluorescence. This may be accounted for by the classical model of the binding of small molecules to protein in which P is dependent on the ratio bound to unbound molecules. We have been able to estimate the quenching due to binding and construct a Scatchard plot. We estimated a wavelength shift from in vitro data consistent with the dependence of P on wavelength seen in our cell work. Generally, the distributions of P are symmetrical. Photon statistics broadens the P distribution of dim cells. However, structure does develop in the P distribution when the cells are deprived of calcium or incubated in the cold. This appears as a shoulder on the P distribution or resolves into two peaks. Calcium deprivation may differentially affect a subpopulation of cells whose significance remains to be explored in various cell types.

Animals↗
PubMed1990

Glycosaminoglycans of bovine aorta endothelial cells: identification and localization by use of a platelet factor 4-fluorescein probe.

We utilized platelet factor 4 (PF4) conjugated to fluorescein to stain the proteoglycans of permeabilized fixed bovine aorta endothelial cells in monolayer culture. Treatment of the monolayers with chondroitin ABC lyase and/or a preparation from Flavobacterium heparinum was used to remove chondroitin sulfate and/or heparan sulfate before staining, with resultant separate identification and partial localization of these glycosaminoglycans. When PF4-fluorescein was utilized with untreated control monolayers, fairly uniform reticular, perinuclear, and cell surface fluorescence was seen. After treatment with chondroitin ABC lyase, fluorescence was retained only on the cell surface. In contrast, treatment with the F. heparinum preparation resulted in the loss of all cell surface fluorescence. Use of both glycosaminoglycan lyases together resulted in loss of essentially all the fluorescence. The cell surface heparan sulfate observed by fluorescence after removal of cell surface chondroitin sulfate appeared to be unevenly distributed, with a heavier accumulation at one pole of each cell. This technique offers a specific method for identification and partial localization of cell surface heparan sulfate.

Animals↗
PubMed1987

Vasoactive effects of buflomedil assessed by fluorescein angiography.

The vascular effects of buflomedil were studied by retinal fluorescein angiography carried out before (control) and after the intravenous infusion of buflomedil (250 mg in 15 min) in patients with different pathological conditions of retinal circulation. Sequential photograms, taken every 36 sec from the 10th sec after the start of the fluorescein injection, showed differences in circulation times between the two sequences in 9 (75%) out of 12 patients. Arterial phase and early venous phase differences were evident, with a mean delay after buflomedil of 3.24 +/- 2.35 sec and 3.22 +/- 2.58 sec, respectively. Significant differences were observed in the time to venous filling completion, with a mean delay of 6.99 +/- 5.19 sec after buflomedil. It is considered that the delay in the retinal arterial phase is related to the arm-retina circulation time, and the slowing down in venous filling is related to a lengthened circulation in the capillary bed.

Adult↗
PubMed2004

Fluorescein permeability and electrical resistance of human skin during low frequency ultrasound application.

Transdermal drug delivery offers an alternative to injections and oral medication but is limited by the low skin permeability of most drugs. The use of low-frequency ultrasound over long periods of time, typically over an hour, has been shown to enhance skin permeability, a phenomenon referred to as sonophoresis. In this study, we investigated the effects of short time sonication of human skin at 20 kHz and at variable intensities and duty cycles on the dynamics of fluorescein transport across the skin (permeability) as well as the changes in the skin's structural integrity (electrical resistance). We found that a short application of ultrasound enhanced the transport of fluorescein across human skin by a factor in the range of 2-9 for full thickness skin samples and by a factor in the range of 2-28 000 for heat-stripped stratum corneum samples (however, samples with very high (10(3)) enhancement were likely to have been damaged by ultrasound). The electrical resistance of the skin decreased by an average of 20% for full thickness samples and 58% for stratum corneum samples. Increasing the duty cycle from 10 to 60% caused a significant increase in permeability enhancement from 2.3 to 9.1, and an increase in intensity from 8 to 23 mW cm(-2) induced a significant increase in permeability enhancement from 2 to 7.4, indicating a clear dependence of the permeability on both duty cycle and intensity. The increase in solute flux upon ultrasound exposure was immediate, demonstrating for the first time the fast response dynamics of sonophoretic enhancement. In addition, a quantitative analysis of the thermal and convective dispersion effects associated with ultrasound application showed that each contributes significantly to the overall permeability enhancement observed.

Administration, Cutaneous↗
PubMed1994

Intraoperative assessment of aneurysm clip placement by intravenous fluorescein angiography.

Clip occlusion of intracranial aneurysms can be difficult, and intraoperative misadventures are still fairly frequent. Many complications are caused by improper clip placement. Intraoperative conventional arteriography is time consuming and requires expensive image-intensifying equipment. As an alternative, we have found that the bolus intravenous administration of fluorescein produces the necessary intra-arterial dye concentration for the direct visualization of the perforating medium and large arteries in proximity to a clipped aneurysm. The technique is straightforward, requiring little additional operating time and an inexpensive set of disposable optical filters. No adverse reactions to fluorescein have been observed.

Cerebral Angiography↗
PubMed1994

Gas-phase microsequencing of peptides and proteins with a fluorescent Edman-type reagent, fluorescein isothiocyanate.

A fluorescent Edman-type reagent, fluorescein isothiocyanate (FITC), was adapted to a gas-phase sequencer fitted with a miniature reaction chamber. Fluorescein thiohydantion amino acids were identified by on-line gradient HPLC with fluorescence monitoring. The optimized protocol for the coupling reaction using FITC and phenylisothiocyanate, and the degree of washing to remove the excess reagents were established. Myoglobin (5 pmol) and lysozyme (5 pmol) were analyzed by the sequencer to obtain a 35-51% initial yield (I.Y.) and 82-88% repetitive yield (R.Y.) for the former, and 51-66% I.Y. and 91-92% R.Y. for the latter. These figures permitted 20-25 amino acid residues to be identified from the N-terminus of 5 pmol samples applied to the sequencer.

Amino Acid Sequence↗
PubMed2004

A new protein conformation indicator based on biarsenical fluorescein with an extended benzoic acid moiety.

We demonstrate herein a new protein conformation indicator based on biarsenical fluorescein with an extended benzoic acid moiety. The present indicator is reactive to a genetically introduced tetracysteine motif (Cys-Cys-Xaa-Xaa-Cys-Cys, where Xaa is a noncysteine amino acid) of proteins. Compared to the original biarsenical fluorescein (FlAsH) and the biarsenical Nile red analogue (BArNile), the present indicator exhibited larger fluorescence intensity changes in response to Ca(2+)-induced conformational rearrangements of calmodulin. A calculation of the highest occupied molecular orbital (HOMO) level of the benzoic acid moiety of the indicator molecule supports possible involvement of a photoinduced electron transfer (PET) process. These results indicate that the present indicator is useful for sensitive detection of protein conformational changes.

Benzoic Acid↗
PubMed1985

Conference on insulin pump therapy in diabetes. Multicenter study of effect on microvascular disease. Assessment of fluorescein angiograms.

Quantitation of the earliest changes in abnormal retinal morphology using fluorescein angiography is potentially superior to retinal photography. However, the critical importance of flawless technique, limitations in the size of the field available for detailed study, and observer variability constitute major disadvantages. A protocol describing standards of photography and of injection was developed. Methodology for counting microaneurysms (Ma) was developed at a central laboratory and applied in suitable photographs obtained at 0 (baseline), 4, and 8 mo in 68 patients. Counts of "definite" or "possible" Ma were made on films projected under standard conditions by two observers known to achieve consistently reproducible results. Semiquantitative assessment of diffusibility of fluorescein reflecting capillary leakage was performed in 61 patients. Leakage was graded according to three degrees of severity permitting study of observer variation, concordance of change in pairs of eyes, and treatment effects. The average number of Ma at baseline was slightly higher in the continuous subcutaneous insulin infusion (CSII) group than in the conventional insulin treatment (CIT) group, but the difference was not statistically significant at the 0.05 level. At both 4 and 8 mo, definite Ma were more prevalent in the CSII group, and the difference was statistically significant at the 0.05 level using both parametric and nonparametric tests. In 27 CSII and 23 CIT patients having complete sets of 0, 4, and 8-mo photographs, Ma counts increased during the 0-4-mo interval. During the 4-8 mo interval, a further increase occurred in the CSII group but contrasted with a decrement observed in the CIT group, which showed no net change from baseline.(ABSTRACT TRUNCATED AT 250 WORDS)

Aneurysm↗
PubMed1981

Inhibition of housefly oxidative detoxication by phthaleins, fluoresceins and related compounds.

1. Phenolphthalein, halogenated fluoresceins, and other triphenylmethane and diphenylmethane derivatives inhibited biphenyl hydroxylation, aldrin epoxidation and several O-dealkylations in insect abdomen homogenates. Phenolphthalein and eosin (50 muM) were 2-3 times more effective than SKF 525-A and piperonyl butoxide (50 muM) as inhibitors of biphenyl hydroxylation in vitro. 2. The phthaleins, Aurin and Aluminon, inhibited both epoxidation and hydroxylation to similar extents, but fluoresceins, Rhodamine B, Malachite Green, and basic diphenylmethane derivatives preferentially inhibited hydroxylation. 3. Tetrabromophenolphthalein ethyl ester and bis-(N-dimethyl-4-aminophenyl-methane inhibited biphenyl hydroxylation in vivo. Bis-(N-dimethyl-4-aminophenyl) methane synergized the toxic effects of 1-naphthyl N-methylcarbamate in live houseflies.

Abdomen↗
PubMed1993

Fluorescein angiography as a means of assessing retinal vascular pathology in oxygen-exposed newborn rats.

Recently a new technique was reported in which examination of the retina of oxygen reared newborn rats was made using indirect fluorescence microscopy (Invest. Ophthal. Vis. Sci. 31:810). We have now improved this technique and systematically employed it as a means of studying oxygen-induced retinopathy in the newborn rat as an animal model for retinopathy of prematurity (ROP). Immediately after birth, litters of pigmented rats were exposed to an elevated oxygen atmosphere that varied in a cyclic fashion between 40% and 80% every 12 hours for 14 days. Rats raised simultaneously in room air served as controls. At the end of the treatment period, rats were deeply anesthetized, their pupils dilated, and 100 microliters of 3% sodium fluorescein was administered via tail venipuncture. After angiography, rats were awakened and returned to their mothers in room air for future assessment. Angiography was performed again at 4, 7, 38 or 56 days after oxygen treatment. Our results suggest that retinal pathology in the rat model parallels that of the human infant with ROP with respect to loss of patent retinal blood vessels in oxygen and subsequent abnormal vasoproliferation upon return to room air. Angiograms performed on room air-raised control rats at 14 days of age demonstrated our ability to resolve the smallest retinal capillaries. Angiograms conducted at the same age in oxygen-exposed rats revealed extensive areas of retinal non-perfusion and substantial arterio-venous shunting between adjacent primary vessels. If rats were maintained in room air for 4 or 7 days following oxygen exposure, a variety of pathologic signs became obvious including: 1) tortuosity of primary vessels mimicking human "plus disease", 2) frequent abnormal budding of tertiary vessels and capillaries, 3) retinal fluorescein leaks indicative of hemorrhages and 4) abnormal capillary tufts not evident at the time of removal from oxygen. By 38 and 56 days post-exposure, all four of these phenomena were still observed. Many of these pathologic phenomena could not have been observed with conventional methods currently used for retinal vascular assessment in small animals.

Animals↗