scholarly journals The biosynthesis in vitro of keratan sulphate in bovine cornea

1972 ◽  
Vol 128 (2) ◽  
pp. 205-213 ◽  
Author(s):  
C. J. Handley ◽  
C. F. Phelps
Keyword(s):  
Uronic Acid ◽  
Cetylpyridinium Chloride ◽  
Corneal Stroma ◽  
Turnover Rates ◽  
Keratan Sulphate ◽  
Tissue Content ◽  
Dowex 1

1. Bovine corneas were incubated in vitro with [U-14C]glucose. 2. The glycosaminoglycans of corneal stroma were isolated and fractionated on cetylpyridinium chloride–cellulose columns. The major components were keratan sulphate (71%), chondroitin 4-sulphate (17%) and chondroitin 6-sulphate (4%). 3. The acid-soluble nucleotides and intermediates of glycosaminoglycan biosynthesis of corneal stroma were separated on Dowex 1 (formate form) and the tissue content and cellular concentrations were determined. 4. The rates of synthesis of the intermediates of glycosaminoglycan biosynthesis in corneal stroma were determined. 5. The incorporation of radioactivity from [U-14C]glucose into the uronic acid and hexosamine components of the glycosaminoglycans present in corneal stroma were measured and the turnover rates of these polymers were calculated. It was found that keratan sulphate was turning over in about 723h and chondroitin 6-sulphate in 251h.

scholarly journals The biosynthesis in vitro of chondroitin sulphate in neonatal rat epiphysial cartilage

1972 ◽  
Vol 126 (2) ◽  
pp. 417-432 ◽  
Author(s):  
C. J. Handley ◽  
C. F. Phelps
Keyword(s):  
Hyaluronic Acid ◽  
Chondroitin Sulphate ◽  
Cetylpyridinium Chloride ◽  
Neonatal Rat ◽  
Turnover Rates ◽  
Glycosaminoglycan Synthesis ◽  
Epiphysial Cartilage ◽  
Sulphated Glycosaminoglycans ◽  
Tissue Contents

1. A system is described, which was used to incubate neonatal rat epiphysial cartilage in vitro with [U-14C]glucose and [35S]sulphate. 2. The acid glycosaminoglycans of neonatal rat epiphyses were extracted and fractionated on cetylpyridinium chloride–cellulose columns. The major components were chondroitin 4-sulphate (65%), chondroitin 6-sulphate (15%), hyaluronic acid (4%) and keratan sulphate (2%). 3. The acid-soluble nucleotides and intermediates of glycosaminoglycan synthesis were separated on a Dowex 1 (formate) system. The tissue contents and cellular concentrations of these metabolites were determined. 4. The rates of synthesis of UDP-glucuronic acid and UDP-N-acetyl-hexosamine from [U-14C]glucose were found to be 0.79±0.16 and 3.2±0.08nmol/min per g wet wt. respectively. 5. The incorporation of [U-14C]glucose into the uronic acid and hexosamine moieties of the polymers was also measured and the turnover rates of the glycosaminoglycans were calculated. It was found that chondroitin sulphate was turning over in about 70h and hyaluronic acid in about 120h. 6. The relative rates of synthesis of the sulphated glycosaminoglycans were calculated from [35S]sulphate incorporation and were found to be in good agreement with those obtained from [U-14C]glucose labelling.

scholarly journals The glycosaminoglycans of human tracheobronchial cartilage

1970 ◽  
Vol 120 (4) ◽  
pp. 777-785 ◽  
Author(s):  
R. M. Mason ◽  
F. S. Wusteman
Keyword(s):  
Molecular Weight ◽  
Potassium Hydroxide ◽  
Chondroitin Sulphate ◽  
Cetylpyridinium Chloride ◽  
Exchange Chromatography ◽  
Chemical Heterogeneity ◽  
Keratan Sulphate ◽  
Sulphate Content ◽  
Age Dependent ◽  
Dowex 1

1. The glycosaminoglycans of human tracheobronchial cartilages from subjects of various ages were liberated by proteolysis of the tissue and purified by ion-exchange chromatography. Purified glycosaminoglycans were fractionated on Dowex 1 resin and cetylpyridinium chloride was used to separate chondroitin sulphates and keratan sulphates occurring in the same fraction. 2. The total chondroitin sulphate content of the cartilages decreased linearly with increasing age. Age-dependent changes in the chemical heterogeneity of chondroitin sulphate were observed, a low-sulphated compound making up 25% of the total glycosaminoglycan at birth but rapidly diminishing in content during the first 6 months of life. Of the total chondroitin sulphate the 6-isomer became rather more prominent than the 4-isomer with increasing age. 3. The total keratan sulphate content of the cartilages increased from trace amounts only at birth to a plateau value by the beginning of the fifth decade. Of the total keratan sulphate approx. 70% was due to a high-molecular-weight compound with a sulphate/hexosamine ratio of 1.5–1.8: 1.0. The degree of sulphation varied between compounds isolated from different individuals. The remaining 30% of the keratan sulphate appeared to be intimately associated with chondroitin 6-sulphate and could only be separated from it after treatment with 0.45m-potassium hydroxide. The hybrid glycosaminoglycans were of lower molecular weight and had a lower sulphate/hexosamine ratio than the major keratan sulphate compound.

Immunolocalization of keratan sulphate in the human embryonic cornea and other human foetal organs

1986 ◽  
Vol 80 (1) ◽  
pp. 181-191
Author(s):  
L. Hyldahl ◽  
R. Aspinall ◽  
F.M. Watt
Keyword(s):  
Stromal Cells ◽  
Corneal Endothelium ◽  
Corneal Stroma ◽  
Cell Type ◽  
Keratan Sulphate ◽  
Suprarenal Glands ◽  
Embryonic Cartilage ◽  
Yolk Sack ◽  
Foetal Intestine

An antibody raised against keratan sulphate was used to localize keratan sulphate in the human embryonic cornea and other human foetal organs. It was found that keratan sulphate was present in the corneal endothelium, as well as in the corneal stroma. In contrast, no keratan sulphate could be demonstrated in the corneal epithelium or in any posterior parts of the human foetal eye. Human corneal stromal cells grown in monolayers in tissue culture lost their capacity to bind this antibody. This suggests that explantation in vitro decreases or alters the expression of keratan sulphate in this cell type. Keratan sulphate was found to be present in abundant quantities in human embryonic cartilage and to a lesser extent in placenta. In contrast, keratan sulphate could not be detected in the foetal intestine, liver, suprarenal glands, the umbilical cord or in the foetal yolk sack.

Collagen fibrillogenesis in vitro: interaction of types I and V collagen

1986 ◽  
Vol 44 ◽  
pp. 210-211
Author(s):  
Arthur J. Wasserman ◽  
Kathy C. Kloos ◽  
David E. Birk
Keyword(s):  
Type I Collagen ◽  
Corneal Stroma ◽  
Minor Component ◽  
Homogeneous Distribution ◽  
Type I ◽  
Type V Collagen ◽  
Fibril Morphology ◽  
Type V ◽  
In Vitro Interaction

Type I collagen is the predominant collagen in the cornea with type V collagen being a quantitatively minor component. However, the content of type V collagen (10-20%) in the cornea is high when compared to other tissues containing predominantly type I collagen. The corneal stroma has a homogeneous distribution of these two collagens, however, immunochemical localization of type V collagen requires the disruption of type I collagen structure. This indicates that these collagens may be arranged as heterpolymeric fibrils. This arrangement may be responsible for the control of fibril diameter necessary for corneal transparency. The purpose of this work is to study the in vitro assembly of collagen type V and to determine whether the interactions of these collagens influence fibril morphology.

scholarly journals Degradation of keratan sulphate by β-N-acetylhexosaminidases A and B

1981 ◽  
Vol 193 (3) ◽  
pp. 811-818 ◽  
Author(s):  
T Ludolph ◽  
E Paschke ◽  
J Glössl ◽  
H Kresse
Keyword(s):  
Isoelectric Focusing ◽  
Human Liver ◽  
Thermal Inactivation ◽  
Sandhoff Disease ◽  
Chromogenic Substrate ◽  
Polymeric Substrate ◽  
Keratan Sulphate ◽  
Human Enzyme ◽  
Tay Sachs Disease

Enzymic cleavage of beta-N-acetylglucosamine residues of keratan sulphate was studied in vitro by using substrate a [3H]glucosamine-labelled desulphated keratan sulphate with N-acetylglucosamine residues at the non-reducing end. Both lysosomal beta-N-acetylhexosaminidases A and B are proposed to participate in the degradation of keratan sulphate on the basis of the following observations. Homogenates of fibroblasts from patients with Sandhoff disease, but not those from patients with Tay–Sachs disease, were unable to release significant amounts of N-acetyl[3H]glucosamine. On isoelectric focusing of beta-N-acetylhexosaminidase from human liver the peaks of keratan sulphate-degrading activity coincided with the activity towards p-nitrophenyl beta-N-acetylglucosaminide. A monospecific antibody against the human enzyme reacted with both enzyme forms and precipitated the keratan sulphate-degrading activity. Both isoenzymes had the same apparent Km of 4mM, but the B form was approximately twice as active as the A form when compared with the activity towards a chromogenic substrate. Differences were noted in the pH–activity profiles of both isoenzymes. Thermal inactivation of isoenzyme B was less pronounced towards the polymeric substrate than towards the p-nitrophenyl derivative.

scholarly journals Relative rates of biosynthesis of collagen type I, type V and type VI in calf cornea

1991 ◽  
Vol 274 (2) ◽  
pp. 615-617 ◽  
Author(s):  
P Kern ◽  
M Menasche ◽  
L Robert
Keyword(s):  
Type I Collagen ◽  
Collagen Type ◽  
Corneal Stroma ◽  
Collagen Type I ◽  
Type I ◽  
Type Vi Collagen ◽  
Sds Page ◽  
Proline Incorporation ◽  
Type V

The biosynthesis of type I, type V and type VI collagens was studied by incubation of calf corneas in vitro with [3H]proline as a marker. Pepsin-solubilized collagen types were isolated by salt fractionation and quantified by SDS/PAGE. Expressed as proportions of the total hydroxyproline solubilized, corneal stroma comprised 75% type I, 8% type V and 17% type VI collagen. The rates of [3H]proline incorporation, linear up to 24 h for each collagen type, were highest for type VI collagen and lowest for type I collagen. From pulse-chase experiments, the calculated apparent half-lives for types I, V and VI collagens were 36 h, 10 h and 6 h respectively.

scholarly journals Determination of the maximum inhibitory dilution of cetylpyridinium chloride-based mouthwashes against staphylococcus aureus: an in vitro study

2008 ◽  
Vol 16 (4) ◽  
pp. 275-279 ◽  
Author(s):  
Evandro Watanabe ◽  
Juliane Maria Guerreiro Tanomaru ◽  
Andresa Piacezzi Nascimento ◽  
Fumio Matoba-Júnior ◽  
Mario Tanomaru-Filho ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Cetylpyridinium Chloride ◽  
In Vitro Study ◽  
Vitro Study

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.

scholarly journals Hemidesmosome formation in vitro.

1983 ◽  
Vol 97 (3) ◽  
pp. 849-857 ◽  
Author(s):  
I K Gipson ◽  
S M Grill ◽  
S J Spurr ◽  
S J Brennan
Keyword(s):  
Basal Lamina ◽  
Mammalian Species ◽  
Corneal Stroma ◽  
Adult Rabbit ◽  
Lamina Densa ◽  
Nucleation Sites ◽  
Epithelial Sheets ◽  
Free Media ◽  
Molecular Components

Intact, viable sheets of adult rabbit corneal epithelium, 9 mm in diameter, were prepared by the Dispase II method (Gipson, I. K., and S. M. Grill, 1982, Invest. Ophthalmol. Vis. Sci. 23:269-273). The sheets, freed of the basal lamina, retained their desmosomes and stratified epithelial characteristics, but lacked hemidesmosomes (HD). Epithelial sheets were placed on fresh segments of corneal stroma with denuded basal laminae and incubated in serum-free media for 1, 3, 6, 18, or 24 h. Tissue was processed for electron microscopy, and the number of HD/micron membrane, the number of HDs with anchoring fibrils directly across the lamina densa from them, and the number of anchoring fibrils not associated with HDs were counted. After 6 h in culture, the number of newly formed HD was 82% of controls (normal rabbit corneas), and by 24 h the number had reached 95% of controls. At all time periods studied, 80-86% of HDs had anchoring fibrils directly across the lamina densa from them. Anchoring fibrils not associated with HDs decreased with culture time. These data indicate that the sites where anchoring fibrils insert into the lamina densa may be nucleation sites for new HD formation. Corneal epithelial sheets placed on two other ocular basal laminae, Descemet's membrane and lens capsule, had not formed HDs after 24 h in culture. These two laminae do not have anchoring fibrils associated with them. Rabbit epithelial sheets placed on the denuded epithelial basal lamina of rat and human corneas formed new HDs. Thus, at least in these mammalian species, HD formation may involve some of the same molecular components.

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