[An N-phenyl-1-amino-8-sulfonaphthalene fluorescent probe for blood plasma albumin].
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Anilino Naphthalenesulfonates↗
Biomedical subjects
G E Dobretsov
Publications and source records attributed to G E Dobretsov.
Fluorescent probes based on styrylpyridinium derivatives: optical properties and membrane binding.
Synthesis and optical properties of the new fluorescent probes - styrylpyridinium derivatives - have been described. Various derivatives of styrylpyridinium, e.g. dimethylamino- (DSP-6 and DSP-12), hydroxy- (HSP-14) and sulphoxy-hydroxy- (SHSP-12) compounds show a long-wave absorption band in ethanol at lambda max abs 486, 402 and 383 nm, respectively. The spectra are sensitive to solvent polarity exhibiting negative solvatochromism by 30-40 nm. Moreover, HSP-14 in aprotic solvents has another absorption band around 600 nm, characteristic of the base. The fluorescence of the probes covers the spectral region 480-650 nm, that of HSP-14 being generated only in the absorption region of the main band (402 nm). The absorption, excitation and fluorescence spectra of these compounds in a liposome suspension have been recorded. The binding of probes to liposomes leads to an increase in fluorescence quantum yield as compared to organic solvents (DSP) and water. Energy transfer in the HSP-14----DSP-12 probe pair has been measured in a liposome suspension. The obtained results suggest that the chemical compounds in question may be useful as fluorescent probes. By varying substituents in the compounds, different spectral characteristics can be attained.
Fluorescent Dyes↗
[Evaluation of the degree of sensitization of hay fever patients using a rat thymocyte-fluorescent probe test system].
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Animals↗
The influence of cholesterol incorporation and removal on lipid-bilayer viscosity and electron transfer in rat-liver microsomes.
The incorporation and removal of cholesterol from rat liver microsomes was used as a methodical approach to investigate the molecular organization of microsomal redox-chains. It was shown that the incorporation of cholesterol in microsomes increases and removal of cholesterol decreases lipid bilayer viscosity as indicated from the rate of fluorescent probe-pyrene eximerisation in cholesterol-enriched and cholesterol-depleted microsomes. The increase of membrane viscosity slows down the initial rates and decreases the rate constants of cytochrome b5 reduction by NAD(P)H, whereas the decrease of membrane viscosity enhances the initial rates and increases the rate constants of these reactions. The rates of cytochrome P450 reduction by reduced pyridine nucleotides do not depend on the viscosity of lipid bilayer. The incorporation and removal of cholesterol from microsomes was not followed by any essential changes in the rates of dimethylaniline N-demethylation, aniline p-hydroxylation, p-nitroanisole O-demethylation, oxygen consumption, oxidation of NADH and NADPH. Thus the reduction of cytochrome b5 by NADH and NADPH is the diffusion-dependent reaction in the redox-chains of microsomes only.
Animals↗
[Spatial organization of low density lipoproteins of the human aorta (study using fluorescent probes)].
Low density lipids of the atherosclerosis-unaffected human aorta, unlike plasma lipoproteins belonging to the same class, contain lipid peroxidation products in the superficial layer. The viscosity of the lipid phase of the aorta and blood plasma is the same. In protein of aorta lipoproteins, tryptophan residues are outside the particle whereas in plasma lipoproteins, they are buried deep in the lipid phase. The data on energy transfer to the probes provide evidence in favour of the fact that in aorta lipoproteins, protein is lying on the particle surface in a thicker layer and occupies a 2-times lesser area than in plasma lipoproteins.
Adult↗
Membrane fluorescent probes for revealing lymphocyte population heterogeneity. II. T-and B-lymphocytes of human peripheral blood.
Lymphocytes of human peripheral blood were stained with a fluorescent probe 3-methoxybenzanthrone (MBA) and their fluorescence intensities measured using a cytofluorimeter. The lymphocytes are shown to be heterogeneous in fluorescence intensity. The fluorescence intensity of B-cells is 2-3 times higher than that of T-cells. Some data suggest that null-cells have an intermediate fluorescence intensity. Thus, with MBA it is possible to distinguish T- and B-lymphocytes in man.
Adult↗
A fluorescence study of apolipoprotein localization in relation to lipids in serum low density lipoproteins.
Fluorescence energy transfer studies were carried out on low density lipoproteins (LDL) containing pyrene, in order to investigate their structure. The results indicate that almost all of the LDL tryptophan residues are located in the same surroundings near the surface of the particle and are immersed in the lipid phase 10-20 A below the lipid/water interface. The data do not support a model of protein spikes protruding from the particle surface. Such spikes have been observed in LDL preparations only after long-term storage.
Apolipoproteins↗
[Nature of E. coli K12 resistance to tetracyclines studied by the fluorescent probe method].
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Cell Membrane↗
[Differences in leukocyte membranes detectable by a fluorescent probe in chronic leukemias].
The fluorescent intensities of the fluorescent probe 3-methoxybenzanthrone (MBA) were compared in lymphocytes of normal donors and patients with chronic lympholeukemia (CLL) and in cells of patients with chronic myeloleukemia (CML). The cells were obtained from the peripheral blood by lymphocytopheresis. In CML, the mean fluorescence intensity of a single cell is 2.2 times higher than in donors, the difference is observed in all patients. In CLL, the mean fluorescence of a single cell negligibly differs from normal. Mathematic analysis of histograms of the fluorescent cells has been shown that the cells may be presented as a sum of two cell groups. In CML and CLL, the relative portion of the cells with the higher fluorescence intensity increases. Possible reasons for changes in the probe fluorescence seen in the cells during leukemias that reflect changes in cell membranes are discussed.
Benz(a)Anthracenes↗
[Surface of a human low-density plasma lipoprotein: areas occupied by proteins and lipids].
The distribution pattern of the protein on the surface of low density lipoproteins (LDL) of blood plasma was studied by quantitative estimation of the efficiency of the inductive-resonance energy transfer between the fluorescent probes and between the proteins and probes. The fluorescent probes used were 4-dimethyl-aminochalcone, 6-(2-anthroyl)-hexane acid and 7-(2-anthryl)-heptane acid. The lipids and proteins were found to occupy approximately equal areas on the LDL surface. The average area of the protein layer on the surface of one LDL particle made up to 480 +/- 60 nm2, the layer width was about 1 nm. The protein was rather evenly distributed on th surface of LDL particle without forming large (diameter much greater than 4 nm) plaques.
Apolipoproteins↗
Membrane fluorescent probes for the demonstration of lymphocyte population heterogeneity. I T. and B lymphocytes of mice and rats.
Non-fixed lymphocytes of rats and mice were stained with the membrane fluorescent probe, 3-methoxybenzanthrone. The probe is capable of binding preferentially with lymphocyte membranes and fluoresces in the green spectral region. Microfluorometry of single cells showed that lymphocytes differ in all lymphoid organs and these may be a 3-10-fold variation in fluorescence intensity. Lymphocytes can be divided into two groups according to fluorescence intensity: "bright" and "dim". The proportions of "bright" and "dim" cells were determined in rats and mice for various lymphoid organs in the normal state, after thymectomy and cyclophosphamide treatment, and also after lymphocyte separation on a nylon wool column. In all cases the proportion of bright lymphocytes corresponded to the B-cell content, and the proportion of dim lymphocytes corresponded to the content of T-cells.
Animals↗
[Changes in spatial organization in sarcoplasmic reticulum membranes in rabbits with experimental thyrotoxicosis].
The structure of sarcoplasmic reticulum membranes (SR) of skeletal muscles from normal and thyrotoxic rabbits was studied with the use of the fluorescent probe pyrene. It was found that protein globules in SR preparations of thyrotoxic animals were submerged into the lipid bilayer to a greater extent than in the reticulum of normal animals. Electrophoresis in polyacrylamide gel revealed no detectable differences between SR of normal and thyrotoxic rabbits. The ratio protein/lipid in SR membranes remained 2:1 in rabbits with thyrotoxicosis.
Animals↗
[4-(n-Dimethylaminostyryl)-1-methylpyridinium fluorescence in a living cell].
A fluorescent compound 4-(p-dimethylinostyryl)-1-methylpyridinium (DSM) has been synthesized, having the absorption maximum about 450 nm and the fluorescence maximum about 590 nm in a water solution. A considerable increase of its fluorescence intensity is found in DNA solutions. The binding of DSM with membranes leads to a shift of its fluorescence maximum to 550 nm. Polychromatic properties of DSM (green fluorescence in membranes, rich yellow - in energized mitochondria, red-orange - in nuclei) are found in DSM stained cells. DSM fluorescence is sensitive to changes in the energized state of cells; the uncupler dinitrophenol or respiration inhibitors-cyanide and amital-cause a strong decrease in the DSM fluorescence intensity in mitochondria. It is ascertained that DSM itself has a low toxicity with respect to cell energy: it had no influence on the mobility of Tetrahymena pyriformis during 23 hours after staining. Thus, DSM may be used as a fluorescent probe for live cells.
Animals↗
[Fluorescence probe analysis of plasma lipoprotein levels].
A method was derived for quantitative determination of the content of lipoproteins belonging to different classes with a fluorescent dye, 1-anilinonaphthalene-8-sulfonate. The method suggested is simple and highly sensitive, it allows one to measure plasma lipoprotein concentrations in man and in different animal species including rabbits with alimentary hypercholesterolemia.
Anilino Naphthalenesulfonates↗
[Stoichiometric protein-lipid interactions in liver microsomal membranes measured by the induction-resonance energy transfer method].
The stoichiometry of proteins and lipids in the microsomal membranes of the liver were studied by the induction-resonance energy transfer method. The energy was transferred between the fluorescent tryptophane residues of membrane proteins and fluorescent probes (pyrene and 4-dimethylaminochalcone) located in the microsomal lipid bilayer. It was assumed that the rate of protein fluorescence is correlated with protein mass. All microsomal proteins were arbitrarily divided into two groups, i.e. proteins located at a distance of more ("peripheral" proteins) and less than 30 A from the lipid bilayer surface. The former proteins made up to 14-24% of total microsomal protein, while microsomal ghosts--less than 10%. In the microsomes the bulk of other proteins is located at an average distance of about 19-26 A from the lipid bilayer surface, while the grosts--at about 4-9 A. Treatment of the ghosts with pronase results in approximation of the protein mass locus with the lipid by about 5 A. In this way the generation of ghosts and their treatment with pronase cause a loss of proteins predominantly from the peripheral parts of the microsomal membrane.
Animals↗
[Effect of incorporation and removal of cholesterol on the lipid bilayer viscosity and the rate of oxidative reactions in rat liver microsomal membranes].
The interrelationship between the lipid bilayer viscosity and the rate of oxidative reactions in rat liver microsomal membranes were investigated. The viscosity of the lipid bilayer is increased upon incorporation of cholesterol into the microsomes and is decreased upon its removal. This results in a corresponding decrease and increase of the rate of cytochrome b5 reduction by NAD(P)H. The rates of cytochrome P-450 reduction and oxidation of dimethylaniline, aniline, p-nitroanisol and NAD(P)H, as well as those of oxygen uptake and lipid peroxidation do not depend on the bilayer viscosity. Hence only the reactions of cytochrome b5 reduction by NADH- and NADPH-specific flavoproteins are diffusion-dependent processes in the microsomal oxidation chain.
Animals↗
[Correlation between antidepressant membrane affinity and their effects on neuromediator reuptake].
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Affinity Labels↗
[Differences between lymphocytes revealed by membrane fluorescent probes].
Interaction of fluorescent probes of 3-methoxybenzanthrone (MBA), 4-dimethylaminochalcone (DMC), and of 1-anilinonaphthalene-8-sulfonate (ANS) with suspensions of rat thymocytes and Peyer's patches lymphocytes has been investigated. By means of the MBA, shown earlier to be a maker of B- and T-lymphocytes, it was established that thymus lymphocytes populations consisted mainly of T-cells (95 +/- 3%), and those of Peyer's patch lymphocytes of B-cells (68 +/- 9%). Definition of parameters of binding and fluorescence of ANS and DMC has shown different physical structure of the thymus and Peyer's patch lymphocyte membranes, resp. This is suggested by differences in binding constant of ANS, and in anisotropy of fluorescence of DMC. The difference in the number of binding sites gives evidence in favour of the fact that Peyer's lymphocytes have a greater membrane mass than thymocytes. T- and B-lymphocyte cells are likely to vary in their own structure and in the number of their membranes.
Anilino Naphthalenesulfonates↗