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PubMed1975

Physicochemical and serological characteristics of respiratory virus fluorescein-isothiocyanate conjugates for fluorescent-antibody diagnosis.

Fluorescein-isothiocyanate (FITC) conjugates were prepared by improved methods from standard reference antisera to influenza A and B, mumps, parainfluenza 1, 2, 3, and 4, herpesvirus, respiratory syncytial virus, and adenovirus hexon. The antisera, prepared in a variety of animals, were fractionated three times with selected optimal concentrations of ammonium sulfate and yielded gamma globulins of adequate purity for conjugation with FITC. Conjugates containing optimal fluorescein-to-protein ratios of between 5 and 10 were produced in 2 h by dialysis labeling. Serological titers of each antiserum and conjugate were determined by complement fixation, hemagglutination-inhibition, serum neutralization, and indirect hemagglutination tests where appropriate. When corrected for dilution, the serological titers of the FITC conjugates were identical to those of the starting antisera. The fluorescent-antibody staining titers correlated well with one of the serological parameters of the original serum. The conjugates stained homologous antigens specifically and were free of nonspecific staining at the working dilution. Undesired staining of host cells which was a problem with some of the conjugates produced from sera containing cellular antibodies was removed by absorption with packed cells. These physicochemical and serological findings were then used as a guide in preparing high quality reagents for fluorescent-antibody identification of respiratory viruses.

Adenoviridae↗
PubMed1976

Fluorescein angiography and its prognostic significance in central retinal vein occlusion.

To determine the prognostic value of fluorescein angiograms in central retinal occlusion 75 patients presenting within the first three months of their initial visual symptoms were studied prospectively. The prognosis was found to be best (complete or partial resolution) in eyes with good capillary perfusion, an intact perifoveal capillary arcade, and leakage only from terminal venules and veins. A broken perifoveal capillary arcade and much capillary leakage, even in the absence of obvious capillary closure, were precursors of permanent macular oedema. All eyes with extensive capillary closure developed severe complications such as thrombotic glaucoma and neovascularization (pre-retinal fibrosis), and all became blind. Macular transit time and leakage from arteries and veins were not used as prognostic features. Fluorescein angiography during the first month of the initial visual symptoms was of prognostic value only in those cases with poor prognosis. The three-month angiogram was an accurate indicator of the visual outcome in all cases.

Capillaries↗
PubMed1978

Fuchs's heterochromic cyclitis: a simultaneous bilateral fluorescein angiographic study of the iris.

Twelve patients with Fuchs's heterochromic cyclitis (FHC) were studied with simultaneous bilateral flucrescein angiography of the iris. The flow began a little earlier in the contralateral iris in 4 cases, and simultaneously in both irides in 8 cases. The radial iris vessels were narrow in 7 eyes with FHC and in the contralateral eyes of 2 elderly patients and 1 patient with pigmentary retinal dystrophy and FHC. An ischaemic sector of the iris was seen in 6 eyes with FHC, neovascularisation of the iris in 8 eyes, and fluorescein leakage of the iris vessels was seen in all eyes with FHC. No neovascularisation of the iris occurred in the contralateral eyes, and only minimal fluorescein leakage was seen at the pupillary border of 5 contralateral eyes. The results support the hypothesis of vascular pathomechanism in FHC.

Adolescent↗
PubMed1980

Indirect choroidal tears at the posterior pole: a fluorescein angiographic and perimetric study.

Retinal fluorescein angiographic and visual field studies were performed on 10 patients who had developed indireect choroidal tears, these procedures being repeated at intervals until the retinal disturbances stabilised. Fluorographic investigations revealed that a transient breakdown of the choroidoretinal barrier to fluorescein dye could be detected in cases investigated within 4 days of injury, and also that the late complication of neovascularisation, producing a serous maculopathy, may resolve and good central vision be retained without recourse to photocoagulation therapy. Patients who had visual field examinations performed within a few days of trauma were noted to have dense central scotomata, but some recovery occurred in most cases, although to a variable degree. Field defects away from fixation corresponded to an extent with areas of post-traumatic pigmentary retinopathy, but in some portions of retina with normal background appearances sensitivity was also noted to be reduced. Scotomata corresponding specifically to choroidal tears or nerve fibre bundle defects were not recorded. Improvement of visual acuity may be expected, except in cases where a choroidal tear has involved the fovea or progressive choroidal neovascularisation led to permanent macular damage.

Adolescent↗
PubMed1980

Experimental occlusion of the central artery of the retina. I. Ophthalmoscopic and fluorescein fundus angiographic studies.

Transient experimental occlusion of the central artery of the retina (OCAR), lasting from 15 to 270 minutes, was produced by clamping the artery in the orbit in 63 eyes of rhesus monkeys. Ophthalmoscopic and fluorescein angiographic studies were performed before and during clamping of the artery, as well as periodically after unclamping, for periods of up to 22 weeks. The effects of transient retinal ischaemia on the retina, optic disc, and retinal vascular bed were studied. 89% of the eyes showed a variable amount of residual retinal circulation on angiography during CAR clamping, but this did not exercise any protective action against ischaemic damage. Duration of the ischaemia was the principal factor determining severity of damage. OCAR for up to 98 minutes produced no significant permanent neural damage, but OCAR for 105 minutes or longer produced irreversible permanent neural damage. There was no significant permanent damage to the retinal vascular bed, though a transient fluorescein leakage was seen after OCAR for 2 1/2-3 hours or longer. The findings revealed that the normal red colour of the optic disc represents retinal vascular filling in the surface layer of the disc and not deeper vascular filling. The various factors influencing the retinal circulation and neural damage in OCAR are discussed.

Animals↗
PubMed1984

Experimental carotid occlusion: funduscopic and fluorescein angiographic findings.

A characteristic fundus picture was consistently produced following acute bilateral common carotid artery ligation in mature rats, reminiscent of human carotid occlusive disease. Two days after ligation it consisted of dilatation and tortuosity of retinal veins, blurring and swelling of the optic disc, retinal whitening primarily along the venous distribution, and straightening of retinal arteries. Fluorescein angiography showed hyperfluorescence of the disc, delay in the rate of retinal arterial and venous filling, venous dilatation, disc oedema, disruption of the retinal capillary bed pattern, and late peripapillary staining/leakage. This pattern was not seen in rats which underwent acute unilateral ligation, although some mild changes were seen on fluorescein angiography. The vascular alterations seemed to regress spontaneously within one week. A peripapillary 'halo' and a granular-appearing nerve fibre layer developed later, exclusively in bilaterally ligated animals.

Animals↗
PubMed1985

Autoimmunity in hereditary retinal degenerations. II. Clinical studies: antiretinal antibodies and fluorescein angiogram findings.

Testing by indirect immunofluorescence for the detection of antiretinal antibodies and lymphocyte stimulation for cell-mediated immunity to retinal antigens was performed on blood obtained from 59 patients with retinitis pigmentosa (RP) and 29 without RP who had other types of retinal disease. The results from the patients' immunological studies were correlated in a masked fashion with six parameters of the fluorescein angiogram: disc staining, peripapillary oedema, vascular arcade oedema, macular oedema, and focal vascular staining (late phases), and disc telangiectasia (early phases). Significant correlations for both groups together were found for IgG antiretinal antibody reactivity and macular oedema (p less than 0.038) and disc staining (p less than 0.033). The non-RP retinal disease group had more significant correlations, including IgG antiretinal antibody reactivity with vascular arcade oedema (p less than 0.018), disc staining (p less than 0.018), and peripapillary oedema (p less than 0.023); the RP patients had significant correlation with IgG reactivity and arcade oedema (p less than 0.045). With combinations of IgG, IgM, and lymphocyte reactivity various significant correlations were found with the fluorescein angiogram.

Adult↗
PubMed1985

Changes at the periphery of a lesion in necrotising scleritis: anterior segment fluorescein angiography correlated with electron microscopy.

Anterior segment fluorescein angiography in scleral disease reveals highly characteristic changes in the vasculature of the anterior segment associated with necrotising scleritis. The vaso-obliterative changes discovered in this investigation have been correlated with the histopathology in a patient who had to have tissue replaced. Light and electron microscopy of scleral tissue excised from sites of vascular closure detected by fluorescein angiography peripheral to the scleral defect revealed pathological changes in the absence of inflammatory cell infiltration. These changes included vascular stasis, partial vaso-obliteration, and fibroblastic transformation of scleral fibrocytes in association with intra- and extracellular degradation of the collagenous component of the matrix.

Collagen↗
PubMed1986

Experimental contamination of Minims of fluorescein by Pseudomonas aeruginosa.

Contamination of fluorescein solutions by Pseudomonas aeruginosa has been a concern of ophthalmologists for many years because of the severity of pseudomonas keratitis. Attempts to prevent contamination have been directed at stringent sterility control during manufacture and the introduction of single-dose disposable containers such as Minims. Deliberate contamination of Minims fluorescein with Pseudomonas aeruginosa was attempted. Under conditions likely to be met with in clinical practice the contents remained sterile. However, under extreme conditions of immersion in pure broth culture of Pseudomonas aeruginosa contamination could be achieved. The relevance of these results to clinical practice is discussed.

Disposable Equipment↗
PubMed1987

Low dose fluorescein angiography of the conjunctiva and episclera.

By reducing the dose of injected fluorescein its leakage from conjunctival and episcleral capillaries has been minimised. These vessels have been demonstrated with great clarity, and the venous circulation, previously obscured by extravascular fluorescein, has also been revealed. The anatomy of the anterior segment vessels, and the blood flow within them, has been studied in eight normal subjects. The anterior ciliary arteries feed an anterior episcleral arterial circle that has superficial and deep components. This supplies the anterior conjunctival and episcleral circulations, the limbal arcades, and the iris arterioles. Where the superficial arterial circle is deficient, isolated vessels emerge from the deep segments of the circle to supply the episcleral plexus and conjunctival arterioles. Watershed zones between the anterior and posterior territories of the conjunctival and episcleral circulations overlap. They may fluoresce up to 30 seconds after the anterior ciliary arteries. The scope of this technique and the implications of these findings are discussed.

Adult↗
PubMed1988

Reflow of obstructed capillaries in the maculae of humans with diabetic retinopathy, observed by fluorescein angiography.

As diabetic retinopathy progresses, the non-perfused area in the retina gradually enlarges as the capillaries are obstructed. To elucidate the process of capillary obstruction we examined macular capillaries in 104 diabetics, using fluorescein angiography. The follow-up period was three to eight years. We found that some capillaries in the macular were obstructed, then became unobstructed. Reflow of blood into the macula after the obstruction was observed in nine patients (8.7%). The time between obstruction and reflow varied from two months to four years. There were four cases of simple, three of preproliferative, and two of proliferative retinopathy. Reflow in the capillaries was confirmed as occurring at the same position and in vessels with the same diameter as seen before the obstruction. There was no leakage of the fluorescein dye from the capillary where reflow took place. We tentatively conclude that one of the causes of this phenomenon is a functional obstruction.

Adolescent↗
PubMed1990

Computer assisted analysis of fluorescein videoangiograms.

The efficient analysis of fluorescein videoangiograms requires immediate access to any frame in a video sequence and complete control over the time base during replay. It should also be possible to compare any two frames by rapid subtraction or to subtract any phase of the study from the remaining sequence. A program that performs all these tasks has been written for a low priced desk-top microcomputer. This should facilitate the clinical introduction of fluorescein videoangiography.

Data Display↗
PubMed2002

Improvement of corneal fluorescein staining in post cataract surgery of diabetic patients by an oral aldose reductase inhibitor, ONO-2235.

AIM: While the mechanism in the pathogenesis of diabetic corneal disease is unclear, aldose reductase has been implicated in corneal disease. The effects of an oral aldose reductase inhibitor (ARI) on the ocular surface of diabetic patients after cataract surgery were studied. METHODS: This clinical trial was designed to be randomised, double blinded, and placebo controlled. Pseudophakic patients with diabetes were randomly assigned to treatment with either oral ARI (ONO-2235) (n=12) or placebo (n=9) for 12 weeks. The vital staining of the ocular surface, tear production and clearance, break up time in tears (BUT), corneal and conjunctival sensation, and symptom score before treatments were examined as well as 4, 8, 12 weeks after the administration. Specular microscopic evaluation was also performed. RESULTS: After a 12 week period of oral ARI administration, fluorescein staining scores (from 2.04 (SD 1.12) to 1.46 (1.18); p=0.016), conjunctival sensation (from 1.15 (0.37) to 1.36 (0.31); p=0.0006), and symptom scores (from 5.38 (1.932) to 4.00 (2.07); p=0.0002) recovered significantly. Fluorescein staining of oral ARI administration also decreased compared with placebo (p=0.017). Rose bengal staining, tear clearance, and corneal sensation were improved although this increase was minor. Tear production, BUT, and specular microscopic evaluation of the corneal epithelium and endothelium did not demonstrate a significant change. CONCLUSION: Oral ARI opposes the ocular surface changes caused by diabetes, by recovery of ocular surface sensitivity as demonstrated through an improvement in vital staining.

Administration, Oral↗
PubMed1999

Energy transfer between fluorescein isothiocyanate and propidium iodide--a problem in the estimation of Tpot with the bromodeoxyuridine-DNA flow cytometry technique?

Energy transfer in flow cytometry can occur when two fluorochromes are bound in close proximity (generally within 100 A) and the emission spectrum of one fluorochrome overlaps significantly with the excitation spectrum of the other. The latter criterium is fulfilled for the fluorochromes fluorescein isothiocyanate and propidium iodide and also the former when they, e.g., are used in bromodeoxyuridine - DNA flow cytometry methods. In the present growth kinetic study using this method, we show that energy transfer does take place between fluorescein isothiocyanate and propidium iodide which results in a detected increase in DNA content with 2-3%. Despite the erroneous increase in the obtained DNA content values, this does not seem to have any influence on the calculation of DNA synthesis time and potential doubling time where the DNA content, based on the relative movement principle of the labelled cells, is used.

Adenocarcinoma↗
PubMedDate not supplied

Use of fluorescein for detecting cerebrospinal fluid rhinorrhoea: A safe technique for intrathecal injection.

The use of intrathecal injections of fluorescein has a role in the diagnosis and management of cerebrospinal fluid leaks using endoscopic techniques. The Polymedic 'pencil point' needle has a lower incidence of headache after dural puncture than standard bevelled needles. We describe a safe method of introducing fluorescein into the thecal space.

Cerebrospinal Fluid Rhinorrhea↗
PubMed1990

Vascular and avascular retinae in mammals. A funduscopic and fluorescein angiographic study.

Intraretinal blood vessels are present in some and absent in other vertebrate species, including the mammals. Among the marsupials, both vascular and avascular retinae are seen. We determined the funduscopic appearance of the eye, investigated the functional aspects of ocular blood flow in both types of retina in marsupials and compared our results with known patterns in placental mammals. The Australian polyprotodont marsupials, the Tasmanian devil, Sarcophilus harrisii, and the quoll, Dasyurus viverrinus, together with an American polyprotodont, the Virginia opossum, Didelphis virginiana, demonstrate variable degrees of tapetal differentiation, pigmentation and a very close parallel course of their intraretinal arteries and veins over considerable distances. Using the technique of fluorescein angiography, we found that retinal blood flow in the 3 vascular Australian species commenced with arterial filling. Early venous was seen next, followed by the capillary blush. This unusual sequence of vascular flow differs from that of the arterial-capillary-venous filling seen in placental mammals. This difference is most likely a consequence of the known looped, end artery organisation found within marsupial nervous systems, of which the retinae are a part. The 2 diprotodont marsupials examined, the brushtail possum, Trichosurus vulpecula, and the sugar glider, Petaurus breviceps, possess avascular retinae. Only a small residual tuft of fluorescein-impermeable vessels projects from the optic disc into the vitreous. Interestingly, the structural complexity of the central visual system in diprotodonts all of whom possess avascular retinae) is commonly accepted as being greater than that of the stem polyprotodont line (which possess vascular retinae). If retinal function matches this internal complexity, then retinal avascularity may, as in birds, be associated with superior vision. However, as the retinae of these mammals clearly lack any nutritive mechanisms directly analogous to those in the retinae of, say, birds or the megachiropteran bats, their retinal nutritive pathways remain enigmatic.

Animals↗
PubMedDate not supplied

Vital capillary microscopy of skin areas at the forefoot of diabetic patients using intraarterial injection of Na-fluorescein.

A modified technique of vital capillary microscopy with intraarterial application of Na-fluorescein has been introduced in the study of nutritional skin microcirculation to assess skin microcirculation of different diabetic patients, comprising one group without neurocutaneous complications (group 2; n = 9), one suffering only from neuropathy (group 3; n = 9) and one with trophic skin lesions in the contralateral foot (group 4; n = 8), all without macroangiopathy, compared to healthy controls (group 1; n = 9). Femoroarterial injection of small boli (10 mg) of Na-fluorescein allowed repeated investigation of the dye appearance times (AT) and capillary-filling times of forefoot skin capillaries within small periods of time before, during and after reactive hyperemia. At rest, AT was significantly shorter in patients of group 4 (16.8 +/- 4.4 s; p < 0.05) compared with groups 1-3 (34.3 +/- 12.8; 31.7 +/- 11.7 and 35.9 +/- 15.3 s). Fifteen seconds after the end of arterial occlusion, dye propagation to the skin was markedly accelerated in groups 1-3 (19.8 +/- 14.0; 14.4 +/- 7.6 and 18.7 +/- 10.6 s, respectively; p < 0.001), but prolonged in group 4 (18.4 +/- 7.4 s). After 10 min, the values at rest were reestablished. No differences between the four groups were found concerning capillary density and morphology. It is concluded that the development of skin lesions in diabetic patients without significant macroangiopathy may be favored by hyperperfusion and impaired vasoregulation. Intraarterial dye injection presents a valuable tool to assess dynamic alterations of the microcirculation at the level of skin capillaries.

Adult↗
PubMed1977

Fluorescein angiographic behaviour of iris masses.

28 cysts and tumours involving the iris are studied by means of fluorescein angiography of the anterior segment. Iris cysts usually showed abnormally dilated and tortuous feeding vessels, but only exceptionally did they leak fluorescein. Inflammatory masses presented massive dye leakage from both tumour-involved and distant iris vessels. Benign stromal masses showed normal angiograms or dye leakage limited to the tumour site. Benign pigmented lesions were angiographically always silent, whereas malignant melanomata showed moderate to heavy leakage from the tumour. This leakage occasionally extended to apparently uninvolved iris areas, specially to the pupillary margin. Exceptionally, a malignant melanoma of the iris was angiographically silent. It is concluded that iris angiography is a useful but not error-free method for the evaluation of iris masses.

Adolescent↗