scholarly journals Pregnancy-related changes in rat cervical glycosaminoglycans

1980 ◽  
Vol 192 (1) ◽  
pp. 1-8 ◽  
Author(s):  
A M Golichowski ◽  
S R King ◽  
K Mascaro
Keyword(s):  
Hyaluronic Acid ◽  
Gestational Age ◽  
Uronic Acid ◽  
Heparan Sulphate ◽  
Late Pregnancy ◽  
Dermatan Sulphate ◽  
Collagen Fibres ◽  
Cellulose Acetate Electrophoresis ◽  
Pregnant Rats ◽  
Pregnant State

Non-pregnant and pregnant rats of known gestational age were killed at intervals and their uterine cervices were excised and digested with papain. Glycosaminoglycans thus extracted were separated by cellulose acetate electrophoresis and stained with Alcian Blue. Glycosaminoglycans were identified by comparison with standards and by serial degradation with chondroitin ABC lyase, butyl nitrite and leech hyaluronidase. Dermatan sulphate, hyaluronic acid and heparan sulphate were identified and quantitative determined by densitometry. The overall concentration of glycosaminoglycans changed little during pregnancy. A 3-fold total increase in uronic acid paralleled the increase in cervical weight. Hyaluronate content, however, increased 17-fold, and rose from 6% of total glycosaminoglycans in the non-pregnant state to 33% at term. Furthermore, the ratio of hyaluronate to hydroxyproline increased 10-fold. These changes are consistent with an accumulation of hyaluronate in the interstices between collagen fibres, resulting in the softening of this tissue that is seen in late pregnancy.

scholarly journals The synthesis of dermatan sulphate proteoglycans by fetal and adult human articular cartilage

1989 ◽  
Vol 261 (2) ◽  
pp. 501-508 ◽  
Author(s):  
L I Melching ◽  
P J Roughley
Keyword(s):  
Hyaluronic Acid ◽  
Articular Cartilage ◽  
Uronic Acid ◽  
Degradation Products ◽  
Chondroitin Sulphate ◽  
Human Articular Cartilage ◽  
Dermatan Sulphate ◽  
Sulphate Proteoglycan ◽  
Adult Human

Non-aggregating dermatan sulphate proteoglycans can be extracted from both fetal and adult human articular cartilage. The dermatan sulphate proteoglycans appear to be smaller in the adult, this presumably being due to shorter glycosaminoglycan chains, and these chains contain a greater proportion of their uronic acid residues as iduronate. Both the adult and fetal dermatan sulphate proteoglycans contain a greater amount of 4-sulphation than 6-sulphation of the N-acetylgalactosamine residues, in contrast with the aggregating proteoglycans, which always show more 6-sulphation on their chondroitin sulphate chains. In the fetus the major dermatan sulphate proteoglycan to be synthesized is DS-PGI, though DS-PGII is synthesized in reasonable amounts. In the adult, however, DS-PGI synthesis is barely detectable relative to DS-PGII, which is still synthesized in substantial amounts. Purification of the dermatan sulphate proteoglycans from adult cartilage is hampered by the presence of degradation products derived from the large aggregating proteoglycans, which possess similar charge, size and density properties, but which can be distinguished by their ability to interact with hyaluronic acid.

scholarly journals Inhibition by heparin-modulated antithrombin III of amidolysis catalysed by mβ-acrosin

1984 ◽  
Vol 217 (3) ◽  
pp. 721-725
Author(s):  
W F Long ◽  
F B Williamson
Keyword(s):  
Hyaluronic Acid ◽  
Molecular Mass ◽  
Antithrombin Iii ◽  
Heparan Sulphate ◽  
Relative Molecular Mass ◽  
Dextran Sulphate ◽  
Dermatan Sulphate ◽  
Arginine Residues ◽  
Cellulose Sulphate

Purified m beta-acrosin catalysed amidolysis of several p-nitroanilides with C-terminal arginine residues. Antithrombin III inhibited amidolysis catalysed by the enzyme. This effect of antithrombin III was potentiated by heparin, and to a modest extent by heparan sulphate, cellulose sulphate, dextran sulphate and xylan sulphate. De-N-sulphated heparin, de-N-sulphated N-acetylated heparin, heparin of low relative molecular mass, chondroitin 4-sulphate, chondroitin 6-sulphate, dermatan sulphate and hyaluronic acid were ineffective.

scholarly journals Synthesis of glycosaminoglycans by human skin fibroblasts cultured on collagen gels

1980 ◽  
Vol 190 (2) ◽  
pp. 243-254 ◽  
Author(s):  
J T Gallagher ◽  
N Gasiunas ◽  
S L Schor
Keyword(s):  
Hyaluronic Acid ◽  
Salt Concentration ◽  
Heparan Sulphate ◽  
Collagen Gel ◽  
Collagen Gels ◽  
Human Skin Fibroblasts ◽  
Dermatan Sulphate ◽  
Sulphated Glycosaminoglycans ◽  
Sulphated Glycosaminoglycan ◽  
Spent Media

A comparison has been made of the synthesis of glycosaminoglycans by human skin fibroblasts cultured on plastic or collagen gel substrata. Confluent cultures were incubated with [3H]glucosamine and Na235SO4 for 48h. Radiolabelled glycosaminoglycans were then analysed in the spent media and trypsin extracts from cells on plastic and in the medium, trypsin and collagenase extracts from cells on collagen gels. All enzyme extracts and spent media contained hyaluronic acid, heparan sulphate and dermatan sulphate. Hyaluronic acid was the main 3H-labelled component in media and enzyme extracts from cells on both substrata, although it was distributed mainly to the media fractions. Heparan sulphate was the major [35S]sulphated glycosaminoglycan in trypsin extracts of cells on plastic, and dermatan sulphate was the minor component. In contrast, dermatan sulphate was the principal [35S]sulphated glycosaminoglycan in trypsin and collagenase extracts of cells on collagen gels. The culture substratum also influenced the amounts of [35S]sulphated glycosaminoglycans in media and enzyme extracts. With cells on plastic, the medium contained most of the heparan sulphate (75%) and dermatan sulphate (> 90%), whereas the collagenase extract was the main source of heparan sulphate (60%) and dermatan sulphate (80%) from cells on collagen gels; when cells were grown on collagen, the medium contained only 5-20% of the total [35S]sulphated glycosaminoglycans. Depletion of the medium pool was probably caused by binding of [35S]sulphated glycosaminoglycans to the network of native collagen fibres that formed the insoluble fraction of the collagen gel. Furthermore, cells on collagen showed a 3-fold increase in dermatan sulphate synthesis, which could be due to a positive-feedback mechanism activated by the accumulation of dermatan sulphate in the microenvironment of the cultured cells. For comparative structural analyses of glycosaminoglycans synthesized on different substrata labelling experiments were carried out by incubating cells on plastic with [3H]glucosamine, and cells on collagen gels with [14C]glucosamine. Co-chromatography on DEAE-cellulose of mixed media and enzyme extracts showed that heparan sulphate from cells on collagen gels eluted at a lower salt concentration than did heparan sulphate from cells on plastic, whereas with dermatan sulphate the opposite result was obtained, with dermatan sulphate from cells on collagen eluting at a higher salt concentration than dermatan sulphate from cells on plastic. These differences did not correspond to changes in the molecular size of the glycosaminoglycan chains, but they may be caused by alterations in polymer sulphation.

scholarly journals Composition and distribution of glycosaminoglycans in cultures of human normal and malignant glial cells

1978 ◽  
Vol 172 (3) ◽  
pp. 443-456 ◽  
Author(s):  
B Glimelius ◽  
B Norling ◽  
B Westermark ◽  
Å Wasteson
Keyword(s):  
Hyaluronic Acid ◽  
Malignant Glioma ◽  
Membrane Fraction ◽  
Heparan Sulphate ◽  
Glioma Cells ◽  
Gel Chromatography ◽  
Total Production ◽  
Dermatan Sulphate ◽  
Sulphated Glycosaminoglycans ◽  
Malignant Glioma Cells

The glycosaminoglycans of human cultured normal glial and malignant glioma cells were studied. [35S]Sulphate or [3H]glucosamine added to the culture medium was incorporated into glycosaminoglycans; labelled glycosaminoglycans were isolated by DEAE-cellulose chromatography or gel chromatography. A simple procedure was developed for measurement of individual sulphated glycosaminoglycans in cell-culture fluids. In normal cultures the glycosaminoglycans of the pericellular pool (trypsin-susceptible material), the membrane fraction (trypsin-susceptible material of EDTA-detached cells) and the substrate-attached material consisted mainly of heparan sulphate. The intra- and extra-cellular pools showed a predominance of dermatan sulphate. The net production of hyaluronic acid was low. The accumulation of 35S-labelled glycosaminoglycans in the extracellular pool was essentially linear with time up to 72h. The malignant glioma cells differed in most aspects tested. The total production of glycosaminoglycans was much greater owing to a high production of hyaluronic acid and hyaluronic acid was the major cell-surface-associated glycosaminoglycan in these cultures. Among the sulphated glycosaminoglycans chondroitin sulphate, rather than heparan sulphate, was the predominant species of the pericellular pool. This was also true for the membrane fraction and substrate-attached material. Furthermore, the accumulation of extracellular 35S-labelled glycosaminoglycans was initially delayed for several hours and did not become linear with time until after 24 h of incubation. The glioma cells produced little dermatan sulphate and the dermatan sulphate chains differed from those of normal cultures with respect to the distribution of iduronic acid residues. The observed differences between normal glial and malignant glioma cells were not dependent on cell density; rather they were due to the malignant transformation itself.

The effect of 1-thyroxine treatment on skin accumulation of acid glycosaminoglycans in primary myxoedema

1986 ◽  
Vol 113 (1) ◽  
pp. 56-58 ◽  
Author(s):  
P. Lund ◽  
K. Hørslev-Petersen ◽  
P. Helin ◽  
H.-H. Parving
Keyword(s):  
Hyaluronic Acid ◽  
Binding Capacity ◽  
Heparan Sulphate ◽  
Dermatan Sulphate ◽  
Water Binding ◽  
Control Value ◽  
Thyroxine Treatment ◽  
Skin Accumulation ◽  
Skin Biopsies ◽  
Acid Glycosaminoglycans

Abstract. We assessed skin accumulation of acid glycosaminoglycans in ten patients with primary myxoedema (median age 70 years) before and during 1-thyroxine treatment (median 7 months). Eight subjects matched for sex and age served as controls. Acid glycosaminoglycans were determined biochemically on small skin biopsies. Hyaluronic acid was elevated in untreated myxoedema, median 0.60, range 0.39–0.90 μg hexosamine/mg dried defatted tissue compared to a median control value of 0.52, range 0.43–0.65 μg hexosamine/mg dried defatted tissue, P < 0.02. Chondroitin-4,6-sulphate and dermatan sulphate showed no elevation, while heparan sulphate was actually reduced in myxoedema, P < 0.01. 1-thyroxine treatment induced a significant reduction in hyaluronic acid, median 0.41, range 0.24–0.71 μg hexosamine/mg dried defatted tissue, while no consistent changes occurred in the remaining three acid glycosamine glycans. The accumulation of hyaluronic acid might contribute to the peculiar non-pitting oedema of the skin in myxoedema, due to the strong water-binding capacity of the substance.

scholarly journals The synthesis of glycosaminoglycans by cultures of rabbit corneal endothelial and stromal cells

1976 ◽  
Vol 158 (3) ◽  
pp. 567-573 ◽  
Author(s):  
B Y J T Yue ◽  
J L Baum ◽  
J E Silbert
Keyword(s):  
Hyaluronic Acid ◽  
Stromal Cells ◽  
Heparan Sulphate ◽  
Deae Cellulose ◽  
Dermatan Sulphate ◽  
Keratan Sulphate ◽  
Monolayer Cultures ◽  
Sulphated Glycosaminoglycans ◽  
Cellulose Column ◽  
Sulphated Glycosaminoglycan

Confluent monolayer cultures of rabbit corneal endothelial and stromal cells were incubated independently with [35S]sulphate and [3H]glucosamine for 3 days. AFter incubation, labelled glycosaminoglycans were isolated from the growth medium and from a cellular fraction. These glycosaminoglycans were further characterized by DEAE-cellulose column chromatography and by sequential treatment with various glycosamino-glycan-degrading enzymes. Both endothelial and stromal cultures synthesized hyaluronic acid as the principal product. The cell fraction from the stromal cultures, however, had significantly less hyaluronic acid than that from the endothelial cultures. In addition, both types of cells synthesized a variety of sulphated glycosaminoglycans. The relative amounts of each sulphated glycosaminoglycan in the two cell lines were similar, with chondroitin 4-sulphate, chondroitin 6-sulphate and dermatan sulphate as the major components. Heparan sulphate was present in smaller amounts. Keratan sulphate was also identified, but only in very small amounts (1-3%). The presence of dermatan sulphate and the high content of hyaluronic acid are similar to the pattern of glycosaminoglycans seen in regenerating or developing tissues, including cornea.

scholarly journals The glycosaminoglycans of canine menisci

1981 ◽  
Vol 197 (2) ◽  
pp. 385-389 ◽  
Author(s):  
M E Adams ◽  
H Muir
Keyword(s):  
Hyaluronic Acid ◽  
Articular Cartilage ◽  
Uronic Acid ◽  
Mechanical Function ◽  
Joint Degeneration ◽  
Dermatan Sulphate

The semilunar menisci of the knee have an important mechanical function and are commonly involved in joint degeneration. However, previously published analyses of the compositions of normal and degenerate human menisci vary widely. In the present study the glycosaminoglycan content and composition of selected areas of the menisci of eight normal knees of working foxhounds were determined. The menisci contained 10% less water and abut 8-fold less glucosaminoglycan than did the articular cartilage of these animals. Although the glycosaminoglycan composition was the same in different regions of the menisci, the total amounts varied considerably. Of the chondroitinase digestible material, approx. 60% was chondroitin 6-sulphate, 25% chondroitin 4-sulphate, 10% chondroitin and 5% dermatan sulphate. Hyaluronic acid accounted for about 6% of the total uronic acid.

Glycosaminoglycans from growing antlers of wapiti (Cervus elaphus)

1997 ◽  
Vol 77 (4) ◽  
pp. 715-721 ◽  
Author(s):  
H. H. Sunwoo ◽  
L. Y. M. Sim ◽  
T. Nakano ◽  
R. J. Hudson ◽  
J. S. Sim
Keyword(s):  
Hyaluronic Acid ◽  
Chondroitin Sulfate ◽  
Cervus Elaphus ◽  
Molecular Size ◽  
Enzymatic Digestion ◽  
Gel Chromatography ◽  
Cellulose Acetate Electrophoresis ◽  
Cartilaginous Tissues ◽  
Chondroitin Sulfates ◽  
Velvet Antler

The emerging wapiti industry in North America is based largely on markets for velvet antlers which are used in oriental medicine. Despite the economic opportunity, enthusiasm has been dampened by incomplete understanding of the chemical and pharmacological properties of velvet antler. This study characterizes polysaccharide constituents of glycosaminoglycans in growing antler of wapiti (Cervus elaphus). Glycosaminoglycans were isolated from four sections (tip, upper, middle and base) of growing antlers, and were studied using cellulose acetate electrophoresis, gel electrophoresis, enzymatic digestion and gel chromatography. The tip and upper sections of the antler which are rich in cartilaginous tissues contained chondroitin sulfate as a major glycosaminoglycan with small amounts of hyaluronic acid. In the middle and base sections containing bone and bone marrow, chondroitin sulfate was also a major glycosaminoglycan with small amounts of hyaluronic acid and chondroitinase-ACI resistant materials. More than half of chondroitin sulfate from the middle and base sections had larger molecular size than did the chondroitin sulfates from the tip and upper sections. Key words: Glycosaminoglycans, chondroitin sulfate, antler, wapiti

scholarly journals Immunochemical analysis of cartilage proteoglycans. Radioimmunoassay of the molecules and the substructures

1980 ◽  
Vol 187 (3) ◽  
pp. 687-694 ◽  
Author(s):  
J Wieslander ◽  
D Heinegárd
Keyword(s):  
Hyaluronic Acid ◽  
Uronic Acid ◽  
Chondroitin Sulphate ◽  
Relative Content ◽  
Binding Region ◽  
Rich Region ◽  
Link Protein ◽  
Specific Radioimmunoassay ◽  
Cartilage Proteoglycans ◽  
Hyaluronic Acid Binding

Antibodies specifically reacting with the link proteins, the hyaluronic acid-binding region and chondroitin sulphate-peptides were used to design specific radioimmunoassay procedures. The sensitivity of the method used for the link protein was about 20 ng/ml, and the other two components could be determined at concentrations of about 2 ng/ml. The radioimmunoassay procedures were tested by using proteoglycan subfractions or fragments thereof. The procedures used to quantify link protein and hyaluronic acid-binding region showed no cross-interference. Fragments of trypsin-digested proteoglycan monomers still reacted in the radioimmunoassay for hyaluronic acid-binding region. Subfractions of proteoglycan monomers separated according to size had a gradually higher relative content of the hyaluronic acid-binding region compared with both chondroitin sulphate-peptides and uronic acid, when the molecules were smaller. The proteoglycans therefore may contain a variably large chondroitin sulphate-rich region, which has a constant substitution with polysaccharide side chains.

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