Heparin and sulfated mucopolysaccharides—a micro system for quantitative differentiation and determination

1977 ◽  
Vol 55 (5) ◽  
pp. 1179-1189 ◽  
Author(s):  
Louis B. Jaques ◽  
Tak K. Sue ◽  
Norman M. McDuffie ◽  
Sandra M. Wice
Keyword(s):  
Aqueous Solution ◽  
Hyaluronic Acid ◽  
Optical Density ◽  
Toluidine Blue ◽  
Linear Relation ◽  
Cetylpyridinium Chloride ◽  
Agarose Gel ◽  
Azure A ◽  
Tissue Extracts ◽  
The Individual

Heparin (Hep), hyaluronic acid, chondroitins (sulfate) A, B, and C, and heparitins (sulfate) A, B, C, and D were subjected to microelectrophoresis in barbital–agarose gel, fixed with cetylpyridinium chloride and stained with toluidine blue. The optical densities of the resulting bands were compared with optical densities obtained upon reaction with azure A in aqueous solution and with the carbazole reagent. A linear relation was obtained between optical density and concentration of purified sulfated mucopolysaccharide (SMP). Less than 1 μg of Hep and 2 μg of other SMPs are required for measurement by electrophoresis, while about 30 μg of each is required with the carbazole reagent. The optical density of a mixture of SMPs was equal to the sum of the densities for the individual SMPs upon microelectrophoresis. It was demonstrated that the individual SMPs in mixtures were distinguished by reaction with specific enzymes and by changes in migration in agarose with barbital, phthalate, ethylenediamine, or propane-diamine buffers, permitting ready demonstration and quantitation of various SMP species. Examples are shown of the application of the procedure to measure the total SMPs and individual SMPs in tissue extracts. The method is sensitive, reproducible, flexible, and measures quantities [Formula: see text] of those measured colorimetrically, yet is relatively unaffected by protein, carbohydrate, or inorganic electrolytes.

Analysis of Heparin – Azure A Metachromasy in Agarose Gel

1972 ◽  
Vol 50 (2) ◽  
pp. 65-71 ◽  
Author(s):  
A. Wollin ◽  
L. B. Jaques
Keyword(s):  
Aqueous Solution ◽  
Salt Concentration ◽  
Low Ph ◽  
Agarose Gel ◽  
Azure A ◽  
Low Salt ◽  
Photometric Measurements

The effect of changes in pH, temperature, salt, and ethanol concentrations on the metachromatic heparin – azure A complex was studied in solution and in agarose gel photometrically. Salt, ethanol, and low pH inhibit the heparin – azure A interaction. With low salt concentration the inhibition is greater in aqueous solution than in agarose gel. Increasing ethanol concentrations interfere stepwise, with a plateau between 40–80% ethanol. Visual estimates of metachromasy do not coincide with the photometric measurements. Increase in temperature decreases the heparin – azure A complex in agarose. The responses to the parameter changes are similar in both media, with differences attributable to the different physical states.

An Improved Microprocedure for the Determination of Heparin

1973 ◽  
Vol 51 (12) ◽  
pp. 994-997 ◽  
Author(s):  
T. K. Sue ◽  
L. B. Jaques
Keyword(s):  
Toluidine Blue ◽  
Quaternary Ammonium ◽  
Cetylpyridinium Chloride ◽  
Agarose Gel ◽  
Complex Samples ◽  
Differential Effects ◽  
Electrophoretic Migration ◽  
Microscope Slides ◽  
Salt Fractionation

The microelectrophoresis method described by Jaques et al. (Can. J. Physiol. Pharmacol. 46, 351–360 (1968)) for identification and quantitation of microquantities of heparin was improved by using the differential effects of inorganic electrolytes on the solubility of quaternary ammonium – mucopolysaccharide complex. Samples were applied to agarose gel coated microscope slides for electrophoresis. The slides were then immersed in 0.1% cetylpyridinium chloride (CPC) in M NaCl at 35 °C with constant agitation for 5 h, washed thoroughly with 0.1% CPC, then stained with toluidine blue. As a result of this treatment, only heparin in concentrations of 0.25–2.0 μg produced optically dense spots on the slides for measurement. Heparin was identified by the combined information from electrophoretic migration, salt fractionation, and metachromatic activity.

Micellar catalysis of organic reactions. VIII. Kinetic studies of the hydrolysis of 2-acetyloxybenzoic acid (aspirin) in the presence of micelles

1982 ◽  
Vol 35 (7) ◽  
pp. 1357 ◽  
Author(s):  
TJ Broxton
Keyword(s):  
Aqueous Solution ◽  
Cetyltrimethylammonium Bromide ◽  
Cetylpyridinium Chloride ◽  
Kinetic Studies ◽  
Base Catalysis ◽  
Plateau Region ◽  
General Base ◽  
Mechanism Of Hydrolysis ◽  
Ph Range ◽  
Hydrolysis Of

The hydrolysis of 2-acetyloxybenzoic acid in the pH range 6-12 has been studied in the presence of micelles of cetyltrimethylammonium bromide (ctab) and cetylpyridinium chloride (cpc). In the plateau region (pH 6-8) the hydrolysis is inhibited by the presence of micelles, while in the region where the normal BAC2 hydrolysis (pH > 9) occurs the reaction is catalysed by micelles of ctab and cpc. The mechanism of hydrolysis in the plateau region is shown to involve general base catalysis by the adjacent ionized carboxy group both in the presence and absence of micelles. This reaction is inhibited in the presence of micelles because the substrate molecules are solubilized into the micelle and water is less available in this environment than in normal aqueous solution.

Inhibin and follistatin concentrations in fetal tissues and fluids during gestation in sheep: evidence for activin in amniotic fluid

1994 ◽  
Vol 141 (2) ◽  
pp. 219-229 ◽  
Author(s):  
S Wongprasartsuk ◽  
G Jenkin ◽  
J R McFarlane ◽  
M Goodman ◽  
D M de Kretser
Keyword(s):  
Amniotic Fluid ◽  
Adrenal Glands ◽  
Late Gestation ◽  
Pituitary Cells ◽  
Fetal Sheep ◽  
Fetal Testis ◽  
Cells In Culture ◽  
Tissue Extracts ◽  
Significant Difference ◽  
The Individual

Abstract The concentrations of inhibin and follistatin in amniotic fluid and in tissue extracts from the placenta, gonads and adrenals of fetal sheep were measured using radioimmunoassays. These tissue extracts were from whole fetuses from days 16 to 45 and from the individual organs from day 46 to 145 (term) and were assayed at multiple dilutions. The capacity of these extracts to alter FSH production of rat anterior pituitary cells in culture was also assessed at multiple dilutions. Immunoactive inhibin concentrations in amniotic fluid from both sexes increased during gestation and levels were significantly greater in males than females. Peak concentrations of immunoreactive inhibin of 11·2±1·9 ng/ml were found in males at 116–125 days of gestation. Follistatin concentrations did not change throughout gestation and no significant difference was noted between sexes. Mean follistatin levels throughout gestation were 3·0±0·9 ng/ml for males and 3·7±0·9 ng/ml for females. Despite the potential for FSH inhibition by inhibin and follistatin, amniotic fluid from both sexes at all stages of gestation stimulated FSH secretion in the pituitary cell bioassays, suggesting the presence of activin which was confirmed by the measurement of immunoactive activin (13·3±2·5 ng/ml) in a specific radioimmunoassay. Maximum concentrations of immunoactive and bioactive inhibin in placental extracts were observed in late gestation (2·2 ±0·6 and 3·8±1·6 ng/g respectively) and there was no significant difference between sexes. Follistatin concentrations in placental cotyledons ranged from 11·5 to 27·1 ng/g with no significant difference between sexes. In view of the higher follistatin concentrations compared with inhibin, it is likely that the capacity of placental extracts to suppress FSH production by pituitary cells in culture is due predominantly to follistatin. Immunoactive inhibin was observed in high concentrations in the fetal testis throughout gestation; with concentrations increasing to a maximum of 1993·0± 519·7 ng/g at 126–135 days of gestation with a ratio of bioactive: immunoactive inhibin of 1:20. Although bioactive and immunoactive inhibin was also observed in fetal ovaries and adrenals from both male and female fetuses, concentrations were lower than those observed in fetal testes. Follistatin concentrations in the fetal testis were elevated between 70 and 95 days (97·6 ng/g) and then declined. Similar concentrations were found in the adrenal glands of both sexes (males 83·5–103·3 ng/g: females 55·3–95·8 ng/g). In both males and females, immunoactive inhibin concentrations in fetal adrenals increased during gestation peaking at levels of 34·4±16·5 and 27·8± 9·0 ng/g respectively. These data suggest that the capacity of adrenal extracts to suppress FSH production by pituitary cells is due to both inhibin and follistatin. These studies demonstrated that significant concentrations of immunoactive inhibin and follistatin are present in amniotic fluid, and the fetal gonads, adrenal glands and placenta in sheep. The role of these proteins during fetal development requires further study. Journal of Endocrinology (1994) 141, 219–229

Removal of toluidine blue from aqueous solution using orange peel waste (OPW)

2014 ◽  
Vol 56 (10) ◽  
pp. 2754-2765
Author(s):  
Ridha Lafi ◽  
Souad Rezma ◽  
Amor Hafiane
Keyword(s):  
Aqueous Solution ◽  
Toluidine Blue ◽  
Orange Peel ◽  
Orange Peel Waste

scholarly journals Определение коэффициента диффузии растворов метиленового синего в дентине зуба человека с помощью спектроскопии отражения и их антибактериальная активность при лазерном воздействии-=SUP=-*-=/SUP=-

2019 ◽  
Vol 126 (6) ◽  
pp. 832
Author(s):  
А.А. Селифонов ◽  
О.Г. Шаповал ◽  
А.Н. Микеров ◽  
В.В. Тучин
Keyword(s):  
Aqueous Solution ◽  
Methylene Blue ◽  
Laser Radiation ◽  
Micellar Solution ◽  
Cetylpyridinium Chloride ◽  
Suppressive Effect ◽  
Human Tooth ◽  
Solution Studies

The work is devoted to the determination of the diffusion coefficients of methylene blue in pure aqueous solution and in a micellar solution of a cationic surfactant in human tooth dentinal sections in vitro using diffuse reflectance optical spectroscopy and the free diffusion model. The determination of the diffusion coefficient of methylene blue for an aqueous solution is (6.74 ± 1.32) • 10−6 cm2/s and (1.93 ± 0.24) • 10−6 cm2/s for methylene blue in micellar solution. Studies of toxicity in daylight in the absence of laser radiation of methylene blue solutions in water and in solution of cetylpyridinium chloride, as well as the photodynamic effect of laser radiation (662 nm) on cells of Candida albicans, Staphylococcus aureus FDA 209P and Lactobacillus were carried out. The effect of laser radiation has a pronounced suppressive effect on all the studied microbial strains.

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