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At least 1,279 records · Page 71Linked to original sources
PubMed1983

Pregnancy-related changes in the mouse oviduct and uterus revealed by differential binding of fluoresceinated lectins.

The binding of 20 fluorescein isothiocyanate (FITC)-labeled lectins to various portions of the pregnant and non-pregnant murine oviduct and uterus was studied by fluorescence microscopy. Five lectins (from Ricinus communis (RCA-I), Maclura pomifera (MPA), Triticum vulgare (wheat germ-WGA), Bauhinia purpurea (BPA), and Ulex europeus (UEA-I] reacted differentially with the epithelium of pregnant as compared with the non-pregnant uterus. The binding of RCA-I, MPA and WGA delineated pregnancy-related changes in the distal oviduct and colliculus tubaris. WGA recognized also pregnancy related changes in the proximal oviduct. The reactivity of the remaining 15 lectins did not distinguish the pregnant and non-pregnant oviduct and uterus, although some of them served to identify specific components of the mouse genital tract. Thus, Soybean lectin (SBA) reacted almost exclusively with the colliculus tubaris. UEA-I alone reacted exclusively with the epithelium of the non-pregnant uterus. RCA-II reacted preferentially with the epithelium of the oviduct and uterus as compared with its weak reactivity with the stroma. Two lectins (from Pisum sativum and Lens culinaris) reacted selectively with stromal cells of the uterus and oviduct. Present data indicate that the differential binding properties of these FITC-labeled lectins can be exploited to identify certain components of the mouse oviduct and uterus and to indicate changes in the cell surface and/or cytoplasm in these structures during pregnancy.

Animals↗
PubMed1982

Characterization of rat colonic cell surface glycoconjugates by fluoresceinated lectins. I. Importance of fixation techniques.

Cryostat and paraffin embedded sections from cecum, proximal and distal colonic segments of male Sherman rats were examined by fluorescence microscopy after labeling with six fluorescein-conjugated lectins. These FITC-conjugated lectins were used as specific probes to define the labeling pattern of carbohydrate containing components of the lumenal and basolateral surfaces of epithelial cells, goblet cell mucin and lumenal mucin at all three sites. Marked regional differences in labeling were detected, indicating that the various carbohydrate components of these cells differ significantly along the length of the colon. Furthermore, the patterns of labeling components with each lectin appeared to vary depending on the fixation technique employed. Cryostat preparations generally resulted in a broader distribution of label and more intense staining with these lectins than fixed paraffin sections. While the reason(s) for these variations remain unclear at this time and will require further studies, the present data emphasize the importance of the fixation method when interpreting results obtained utilizing FITC-conjugated lectins.

Animals↗
PubMed1987

Differential endocytosis of fluorescein iso-thiocyanate-concanavalin A by normal and chronic myeloid leukemic granulocytes.

Isolated granulocytes from normal individuals and patients suffering from chronic myeloid leukemia (CML) displayed different fluorescent patterns on treatment with fluorescein isothiocyanate concanavalin A (Fl-Con A). The ligand was internalized by 86% of the normal granulocytes, while 80% of the leukemic granulocytes exhibited Fl-Con A localized on the cell periphery. In further experiments, pretreatment of the normal granulocytes with cytochalasin B, iodoacetamide, 2-deoxyglucose and sodium fluoride (but not with sodium azide or dinitrophenol) was found to drastically inhibit internalization of the ligand. However, pretreatment of granulocytes from CML patients with cytochalasin B and 2-deoxyglucose, caused only a little alteration in the pattern of Fl-Con A labelling relative to untreated cells. These results indicate that CML granulocytes are defective in their ability to endocytose Fl-Con A. We suggest that this differential interaction between Fl-Con A and normal and leukemic granulocytes is a convenient system to study the initial steps in receptor mediated endocytosis of Concanavalin A.

Azides↗
PubMed1983

Demonstration of specific receptors for fluoresceinated casein on human neutrophils and monocytes using flow cytometry.

Casein is chemotactic for human neutrophils (PMNs) and monocytes. The binding of fluorescein (FITC)-conjugated casein (mixture of alpha, beta, and kappa-casein) and purified alpha-casein to PMNs, monocytes, and lymphocytes was analyzed using flow cytometry. These studies demonstrate that 75-95% of PMNs and 46-85% of monocytes have membrane receptors for casein while lymphocytes lack these receptors. The binding of FITC-casein and FITC-alpha-casein was specific and was blocked only by unlabeled casein and alpha-casein, but not by ovalbumin, bovine or human serum albumin, beta-casomorphin, C5a, or formyl-methionyl-leucylphenylalanine (fMLP). The binding of FITC-casein was reversible when PMNs were stained with this fluorescent agent and subsequently incubated with unlabeled casein. Double-labeling studies of mononuclear cells using FITC-casein and the OKM1 monoclonal antibody in conjunction with a rhodamine conjugated anti-Ig second antibody demonstrate that mononuclear cells binding FITC-casein also stain with the OKM1 monoclonal antibody, indicating a specificity for monocytes.

Binding, Competitive↗
PubMed1985

Further studies on the surface saccharides in Trichomonas vaginalis strains by fluorescein-conjugated lectins.

Fluorescence emitted by individual cells of several Trichomonas vaginalis strains, nearly all of which were cloned, incubated with fluorescein-conjugated lectins in the absence (experimental) or presence (control) of inhibitory sugars, or else in phosphate-buffered saline alone (autofluorescence) was measured with a Leitz MPV Compact microfluorometer. Irrespective of whether the organisms were postfixed in formalin or glutaraldehyde, the relative fluorescence emitted by the cells was closely comparable, provided that appropriate neutral density filters were employed. However, autofluorescence was much higher for glutaraldehyde-fixed trichomonads. Therefore, although better preserved and more amenable to subsequent manipulations, such organisms were found unsuitable for use in "qualitative" titration of the fluorescence emitted by various strains. Provided that the necessary precautions were taken, comparable fluorescence readings were obtained with trichomonads affixed to glass slides by heat (41 degrees C, on a section spreader) or by a cytologic centrifuge (Cytospin 2). Large numbers of concanavalin A (Con A)-binding sites were present on organisms of all strains, irrespective of their virulence for human patients and as estimated by the subcutaneous mouse assay; these sites were shown with the aid of D-mannose to be mannose or mannose-related residues. More binding sites for soybean agglutinin (SBA) were found on the virulent than on avirulent strains. On the basis of inhibition experiments, the sugar residues mainly responsible for these differences appeared to be D-lactose residues. Similar differences were observed with Ricinus communis agglutinin Type I (RCA I), for which D-galactose was employed as the competing sugar. However, with two cloned strains the situation with regard to RCA I binding was reversed - more of the lectin bound to a mild than to a virulent strain. The results obtained with Ricinus communis Type II agglutinin (RCA II) were often similar to those noted for RCA I; however, in most instances the inhibition with N-acetyl-D-galactosamine (GalNAc) was lower. Furthermore, the results noted with the GalNAc-specific agglutinins from Dolichus biflorus and Helix pomatia suggested that only very few GalNAc residues were available for binding on the surfaces of all T. vaginalis strains examined in the course of this study. Statistical analyses of fluorescence emitted by four clones of each, Balt 42 (virulent) and JH31A (avirulent) T. vaginalis strain upon incubation with Con A and SBA revealed homogeneity of these strains with regard to the number of the specific surface saccharide residues, D-mannose and D-lactose.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1990

Selective labeling of alpha-bungarotoxin with fluorescein isothiocyanate and its use for the study of toxin-acetylcholine receptor interactions.

The main product of the reaction of fluorescein isothiocyanate (FITC) and bungarotoxin (Bgt) under near stoichiometric conditions is a monofluorescein derivative preferentially labeled at Lys 26, a highly conserved residue known to be involved in the binding (McDaniel, C.S., Manshouri, T., and Atassi, M.Z. (1987) J. Prot. Chem. 6, 455-461; Garcia-Borron, J.C., Bieber, A.L., and Martinez-Carrion, M. (1987) Biochemistry 26, 4295-4303) of postsynaptic neurotoxins specific for the nicotinic acetylcholine receptor (AcChR). The fluorescently labeled toxin retains a high affinity for the AcChR, and an unaltered specificity. Binding of FITC-Bgt to AcChR results in a significant decrease in the fluorescence intensity of the probe. This AcChR-mediated quenching of FITC-Bgt fluorescence allows for a continuous monitoring of the binding process. The quenching of free and bound FITC-Bgt by charged and neutral quenchers shows few fluorophore accessibility changes as induced by the toxin-bound state. The results are consistent with a model in which the positively charged concave surface of the toxin interacts with a negatively charged complementary surface in the receptor molecule.

Animals↗
PubMed1995

Influence of topical cyclosporine A and dissolvent on corneal epithelium permeability of fluorescein.

The corneal stroma is the major barrier to penetration for the lipophilic Cyclosporine A (CsA) molecule and prevents the use of the common ophthalmic solvents. At present, corn oil, castor oil and olive oil are the three most commonly used vehicles. The aim of this study was to determine the effect that topically applied CsA dissolved in different oils has on corneal epithelial permeability measured by fluorophotometry. Forty healthy volunteers, with absence of ocular or systemic disease and not receiving topical or systemic drugs were enrolled. Measurements were taken before and 45 min after the instillation of 40 microliters of a 2% aqueous solution of sodium fluorescein without preservatives. Basal corneal permeability and the permeability 24 h after the instillation of 2% CsA-olive oil, olive oil alone, 2% CsA-castor oil, castor oil alone, 2% CsA-corn oil and corn oil alone, were calculated. To prepare the topical 2% CsA, a Sandimmun oral solution (Sandoz, Basel, Switzerland) was employed under sterile conditions. We found that epithelial permeability 24 h after the instillation of any CsA formulations or solvents increased more than 6.62 times (p <0.001). No differences in corneal permeability values were found between any of the CsA formulations and the vehicles. We conclude that oils used to dissolve CsA are mainly responsible for the increased corneal epithelial permeability. No differences were found in the effects of the tested solvents on corneal epithelial permeability.

Administration, Topical↗
PubMed1988

Asymmetric incorporation of [14C]cyanate and of fluorescein isothiocyanate in mamillary body of conditioned rats.

A marked decrease in overall learning capacity has been observed in rats injected with cyanate. Therefore it was of interest to test whether learning influenced carbamylation of brain proteins. Incorporation of [14C]cyanate into proteins of the mamillary body was selectively modified following operant conditioning of the rat, so that trained rats showed an asymmetric image with higher levels of incorporation in the right side than in the left side, as compared to control rats. These results were confirmed using fluorescein isothiocyanate. The asymmetry persisted once the learning had been well established.

Animals↗
PubMed1981

Ethanol and acetaldehyde do not increase the blood-brain and blood-retinal barrier permeability to sodium fluorescein.

No extravasation of sodium fluorescein from capillaries of the cerebral cortex and retina could be demonstrated in rats during severe nonlethal ethanol and/or acetaldehyde poisoning. The results do not exclude the possibility that repeated intoxications may gradually open the blood-brain and blood-retinal barriers, but do indicate the absence of an effect during acute intoxication.

Acetaldehyde↗
PubMed1981

Fluorescein-concanavalin A conjugates distinguish between normal and malignant human cells: a preliminary report.

A method is described for using a fluorescein isothiocyanate concanavalin A conjugate to stain human cell membranes in formalin fixed paraffin embedded tissue. 57 neoplastic and normal tissue sites were examined. In 54 malignant tumours, bright green fluorescence was confined to the cell membranes while in 23 benign tumours and normal tissue sites, the membranes were unstained or showed a diminished level of fluorescence. The distinction between malignant and hyperplastic or normal cells was clear cut and definite.

Breast Neoplasms↗
PubMed1984

Binding of fluorescein isothiocyanate conjugated lectins to rat spermatogenic cells in tissue sections. Enhancement of lectin fluorescence obtained by fixation in Bouin's fluid.

The testes from three months old Sprague-Dawley rats were fixed in Bouin's fluid or neutral buffered 10% formalin, embedded in paraffin, sectioned and after deparaffination stained with the following fluorescein isothiocyanate coupled lectins: PNA, WGA, Con A, RCA, SBA, DBA and UEA. The results show that there are considerable differences in the staining pattern of various spermatogenic cells between different lectins. The fixation in Bouin's fluid enhanced the staining of all the lectins compared to formalin fixation in which only a weak staining could be seen in the acrosomes of spermatids after WGA or PNA staining. PNA and WGA stained specifically the acrosome of the developing spermatids, which was seen from the beginning of the acrosome formation and lasted up to late spermiogenesis. However, the staining with PNA decreased in the late spermatids whereas the intensity of the staining remained unchanged with WGA. Con A did not stain the acrosome but stained unspecifically the cytoplasm of all spermatogenic cells. RCA stained faintly the acrosome throughout the spermatid differentiation. DBA and UEA stained specifically the chromosomes of B spermatogonia. DBA also faintly stained the cell membranes of early spermatids. SBA did not show any specific staining of the spermatogenic cells. Based on this it is suggested that the carbohydrates and glycoproteins which are known to be present in the acrosome are formed already in the beginning of the acrosome formation. The decrease in the PNA staining in late spermatids possibly reflects the fact that the receptor molecules are not synthesized in late spermatids but are formed in earlier developmental stages and are thereafter preserved in the acrosome.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Fluorescein endoscopy. A technique for noninvasive assessment of intestinal ischemia.

Late diagnosis contributes significantly to the mortality and morbidity of mesenteric ischemia. Although flexible endoscopy permits noninvasive assessment of the colon, ischemic colitis is often difficult to differentiate from nonspecific proctitis/colitis or mucosal changes seen in inflammatory bowel diseases. Intravenously administered fluorescein sodium (FSC) has long been used intraoperatively to assess bowel viability because its uptake is inversely proportional to the degree of intestinal ischemia. The authors wished to determine if FSC could be used to augment conventional endoscopy in an attempt to identify and monitor ischemic areas of the bowel within reach of the endoscope. Segmental mesenteric ligation of canine rectum was performed, and serial transanal biopsies obtained. Endoscopy was performed after intravenous FSC administration, using a rigid proctoscope, with a long-wave ultraviolet light source inserted into the lumen of the scope. Areas of nonspecific-appearing proctitis did not take up FSC and were easily differentiated from well-perfused areas by their failure to fluoresce. Ischemic areas were monitored in this manner over a 28-day period. Complete resolution of ischemic mucosal changes occurred with late stricture formation. In humans, the sigmoid and left colon are the areas most frequently affected by ischemic colitis. Because long-wave ultraviolet light is not transmitted through glass fibers, a different light source was needed for potential clinical application. To this end, a single fiber of an argon laser was inserted through the instrument channel of a colonoscope, resulting in greater fluorescence than seen with the ultraviolet light. FSC endoscopy appears to be a sensitive and accurate method of detecting and monitoring ischemic colitis and has many potential clinical applications.

Adult↗
PubMed1996

Uptake and distribution of fluorescein-labeled D2 dopamine receptor antisense oligodeoxynucleotide in mouse brain.

To determine the uptake and distribution of oligodeoxynucleotides in brain, a 20-mer phosphorothioated oligodeoxynucleotide complementary to a portion of the D2 dopamine receptor mRNA was fluorescently labeled with fluorescein isothiocyanate (FITC) and injected into the lateral cerebral ventricles of mice. At various survival times after the injection, the brains were removed, fixed, sectioned, and viewed under a fluorescent microscope. The results showed that the oligodeoxynucleotide was rapidly taken up into the brain. Initially the label was relatively diffusely spread throughout the interstitial spaces of the brain, then became redistributed to the cellular compartments. The signal extended from those forebrain nuclei located immediately in contact with the ventricles, such as the corpus striatum, septum, and hippocampus, to areas further removed from the ventricles, such as the cerebral cortex, nucleus accumbens, and substantia nigra. When the FITC-labeled D2 antisense oligodeoxynucleotide was given once daily for 4 d, the signal intensity seen 24 h after the last injection appeared to be of greater intensity overall compared to that seen after a single injection. At early time-points the oligodeoxynucleotide signals appeared to be punctuated and were found in cell bodies as well as in proximal dendritic processes. However, not all cells were equally labeled, suggesting an uneven uptake and accumulation of the D2 antisense into the various cell types. At later time-points the fluorescent signal appeared granular; at these times the injected material was largely degraded. These studies show that a D2 dopamine receptor antisense oligodeoxynucleotide is rapidly taken up from cerebral ventricles into brain, becomes widely distributed throughout the brain tissue to areas far removed from direct contact with the ventricles, and appears to accumulate to a different extent in the various brain areas and cell types.

Animals↗
PubMed1997

Labeling of oligonucleotides with fluorescein.

The labeling of oligonucleotide probes using a fluorescein-labeled nucleotide is described. The reaction is characterized by careful control of the nucleotide and probe molar ratio in order to produce a tail that gives good detection sensitivity without compromising hybridization stringency control of the probe sequence. The labeling reaction can be easily monitored for incorporation of the fluorescent label and the probes can be used in many applications.

DNA↗
PubMed2005

Direct fluorometric analysis of a newly synthesised fluorescein-labelled marker for glomerular filtration rate.

There is an obvious and growing medical need for an accurate determination of kidney function in the diagnosis and management of renal diseases. The glomerular filtration rate (GFR) is the accepted gold standard measurement of kidney function. Several approaches to estimate the GFR are available, but most of them are inconvenient and, therefore, of limited acceptance. A new method of quantification with fluorescein-isothiocyanate (FITC) sinistrin (FS), a novel GFR marker, has been evaluated. The method is based on the fluorescence label of FS and can be performed with a standard fluorometer. To control the interference of protein with the fluorescence signal, a calibration function was developed. The accuracy of the fluorometric method established is comparable to the so-called "gold standard" of enzymatic determination of polyfructosan. Moreover, FS is easy to handle and requires low-cost instruments. Our results demonstrate the potential of the direct fluorometric analysis of the new FITC-labelled marker of being a precise, simple, rapid and cost-effective method for diagnosing disturbed kidney function and monitoring its treatment efficacy. The dramatic decrease in analytical effort will result in a significantly higher acceptability of GFR determination.

Animals↗
PubMed2002

[Pigment epithelium detachments in AMD (age-associated macular degeneration) and "polypoid choroidal vasculopathy". A fluorescein and indocyanine green angiography study].

AIMS: A prospective analysis to differentiate the angiographic appearance of pigment epithelial detachments (PED) in age-related macular degeneration (AMD) was performed. The aim was to verify if characteristics of polypoidal choroidal vasculopathy (PCV) can be observed in elderly patients with Central European background. PATIENTS AND METHODS: Fluorescein and indocyanine green angiography was performed in 101 consecutive patients (53-87 years, 63 females, 38 males) with clinical signs of PED and drusen. RESULTS: Different types of PED could be differentiated: PCV-associated PED in 14 patients (13.9%), vascularized PED in 72 patients (71.2%), and nonvascularized PED in 15 patients (14.9%). CONCLUSIONS: The clinical diagnosis of PED in AMD can be differentiated by angiography into PCV-associated PED, vascularized PED, and nonvascularized PED. This differentiation is important because PCV-associated PED may have a pathogenetic and genetic background other than age-related changes in Bruch's membrane. These patients also may share a natural course and treatment options different from PED in AMD.

Aged↗
PubMed1976

Acute pulmonary edema following intravenous fluorescein angiography.

A 64-year-old white man with no history of cardiac or bronchopulmonary disease developed acute pulmonary edema shortly after undergoing diagnostic intravenous fluorescein angiography. The patient responded quickly to a course of oxygen, positive pressure breathing, diuretics, and sedation.

Fluorescein Angiography↗