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Showing papers on "Keratan sulfate published in 1988"


Journal ArticleDOI
TL;DR: Study of monomers newly synthesized by calf and steer chondrocytes suggests that the age related changes in monomer structure result largely from changes in proteoglycan synthesis or intracellular processing.

94 citations


Journal Article
TL;DR: It is concluded that corneoscleral explant organ culture is a useful tool for extracellular matrix studies within a time window from 7 to at least 14 days in culture.
Abstract: Human corneoscleral explants were maintained for several weeks in defined, serum-free media. Trabecular cell vitality, as judged by vital stain exclusion, is high for at least one month. Trabecular ultrastructure, as compared to that of fresh eyes, first shows minor cellular and extracellular matrix degradation after 3 weeks in culture. The biosynthetic profiles of trabecular glycosaminoglycans (GAGs) change significantly by 3 weeks in culture. Eyes that are stored at 5 degrees C for up to 48 hr postmortem exhibit changes in trabecular ultrastructure and in GAG profiles; both characteristics return to normal by 7 days in culture. The incorporation pattern of 35S-sulfate and 3H-glucosamine into the GAGs of the trabecular meshwork (TM) is distinct from corneal or scleral incorporation. The relative incorporation of 3H-glucosamine into trabecular GAGs, as determined by sequential enzymatic degradation, is: 22.3% hyaluronic acid (HA), 27.9% chondroitin sulfate (CS), 21.3% dermatan sulfate (DS), 5.9% keratan sulfate (KS), 17.7% heparan sulfate (HS) and 4.9% unidentified material. The relative incorporation of 35S-sulfate into trabecular GAGs is: 0% HA, 32.9% CS, 34.8% DS, 7.7% KS, 13.8% HS and 11.1% into unidentified material. This profile is in good agreement with the profile that was previously obtained for human and nonhuman primate meshworks prior to culture. We conclude that corneoscleral explant organ culture is a useful tool for extracellular matrix studies within a time window from 7 to at least 14 days in culture.

94 citations


Journal ArticleDOI
TL;DR: The mode of calcium binding to GAGs is consistent with a recently proposed mechanism of growth plate calcification which states that cartilage proteoglycan functions as a reservoir of calcium for calcification of epiphyseal cartilage.

92 citations


Journal Article
TL;DR: In this paper, the authors found that endothelial cells from fetal and neonate rabbit cornea and endothelium-derived fibroblasts from healing wounds of adult cornea synthesize and deposit type III collagen.
Abstract: Whole neonate rabbit corneas and adult corneas containing 2-week-old scars were incubated in the presence of [14C] glycine. Radiolabeled collagen extracted from the corneas and scar tissue were analyzed by sodium dodecylsulfate/polyacrylamide gel electrophoresis and fluorography to determine the types and relative quantity of collagen polypeptides present and synthesized by these tissues. In addition to other collagen types, type III was found in both neonate cornea and scar tissue from adult cornea, albeit in relatively small quantities. Type III collagen in normal cornea was associated with the residue after pepsin digestion and formic acid extraction of the tissue, and the same type of collagen was extracted from scar tissue after similar treatment. Type III collagen-specific monoclonal antibody bound to developing normal corneas and healing adult tissue sections, as determined by immunofluorescence. Antibody binding was localized to the endothelium and growing Descemet's membrane in fetal and neonate corneas, and restricted to the most posterior region of the corneal scar tissue. Although monoclonal antibody to keratan sulfate, used as a marker for stromal fibroblasts, bound to most of the scar tissue, the antibody failed to bind to the posterior scar tissue positive for type III collagen. We conclude that endothelial cells from fetal and neonate rabbit cornea and endothelium-derived fibroblasts from healing wounds of adult cornea synthesize and deposit type III collagen. Moreover, this collagen appears to be incorporated into the growing Descemet's membrane of normal corneas and narrow posterior portion of the scar tissue.(ABSTRACT TRUNCATED AT 250 WORDS)

72 citations


Journal ArticleDOI
TL;DR: It is concluded that serum keratan sulfate levels rise predictably following acute loss of proteoglycan from a single joint.
Abstract: Following injection of chymopapain into a single knee joint in rabbits, serum keratan sulfate levels rose sharply and remained elevated for at least 48 hours before returning to preinjection levels. These changes were accompanied by depletion of proteoglycans from articular cartilage in the injected joint. We conclude that serum keratan sulfate levels rise predictably following acute loss of proteoglycan from a single joint.

69 citations


Journal Article
TL;DR: Findings support the proposal that the a and c bands were specific binding sites for keratan sulphate proteoglycan (Scott & Haigh, 1985b), and evidence from studies on cornea and cartilage suggests that keratan sulfate, rather than chondroitin sulphate is produced in conditions of O2 lack.
Abstract: Corneas from mouse, rat and rabbit were analysed quantitatively and/or qualitatively for collagen and acid glycosaminoglycans. They were examined by light and electron microscopy, using Alcian blue and Cupromeronic blue, in critical electrolyte concentration methods, with or without digestion by hyaluronidase, chondroitinases and keratanase, for their sulphated glycosaminoglycan distributions. Glycosaminoglycan patterns were very different in the three species. Mouse lacked chemically detectable keratan sulphate, which was present in considerable amounts in rat and rabbit stroma. Mouse corneal stroma proteoglycan filaments were located predominantly at the gap zone of the collagen fibrils, mainly at the d band, with few at the a and c bands. Rat and rabbit micrographs were more complicated, with many proteoglycan filaments at the a and c, as well as the d and e bands. These findings support the proposal that the a and c bands were specific binding sites for keratan sulphate proteoglycan (Scott & Haigh, 1985b). Evidence from studies on cornea and cartilage suggests that keratan sulphate, rather than chondroitin sulphate is produced in conditions of O2 lack. Metabolic mechanisms which could account for this balance are proposed The production of uridine diphosphate glucuronic acid is the key step, which is sensitive to hypoxia, lactate and NAD:NADH ratios.

68 citations


Journal ArticleDOI
TL;DR: Based on differences in the storage material, macular corneal dystrophy appears to manifest heterogeneity with at least two distinct varieties: keratan sulfate negative (type 1) and keratan sulphate positive (type 2).

68 citations


Journal ArticleDOI
TL;DR: The pattern of proteoglycan secretion observed in polarized cell cultures mimicked that observed for uterine cells, although the preferential increase in KSPG production by polarized cells could not be attributed to an estrogen response and provides evidence for a novel influence of cell polarity on the cell's ability to secrete sulfated glycoconjugates.
Abstract: We have studied proteoglycan secretion using a recently developed system for the preparing of polarized primary cultures of rat uterine epithelial cells. To mimic their native environment better and provide a system for discriminating apical from basolateral compartments, we cultured cells on semipermeable supports impregnated with biomatrix. Keratan sulfate proteoglycans (KSPG) as well as heparan sulfate-containing molecules (HS[PG]) were the major sulfated products synthesized and secreted by these cells. The ability of epithelial cells to secrete KSPG greatly increased in parallel with the development of cell polarity. Furthermore, KSPG secretion occurred preferentially to the apical medium in highly polarized cultures. In contrast, HS(PG) secretion did not increase along with development of polarity, although most HS(PG) (85%) were secreted apically as well. Pulse-chase studies indicated that highly polarized cultures secreted 80-90% of the sulfated macromolecules they synthesized, predominantly to the apical secretory compartment. The half-lives for KSPG and HS(PG) secretion were approximately 3 and 4 h, respectively. Parallel studies of cells cultured on tissue culture plastic-coated with biomatrix indicated that neither the state of confluency nor the biomatrix was primarily responsible for inducing the KSPG secretion observed in polarizing cultures. Experiments with uterine strips indicated that the steroid hormone, 17-beta-estradiol, markedly stimulated synthesis and secretion of sulfated macromolecules, but had no preferential effect on KSPG production. The ratio of KSPG to HS(PG) secretion from uterine strips was similar to that found in the apical medium of highly polarized cell cultures. Thus, the pattern of proteoglycan secretion observed in polarized cell cultures mimicked that observed for uterine cells, although the preferential increase in KSPG production by polarized cells could not be attributed to an estrogen response. Collectively, these studies describe the major sulfated molecules secreted by rat uterine epithelial cells under varying conditions and provide evidence for a novel influence of cell polarity on the cell9s ability to secrete sulfated glycoconjugates.

67 citations


Journal Article
TL;DR: Corneal keratan sulfate proteoglycan from scar tissue of experimental penetrating corneal wounds in rabbits was analyzed 2-8 weeks after injury using three previously characterized antibodies and showed identical quantitative binding of antibodies against core protein and KS from normal and scar tissue.
Abstract: Corneal keratan sulfate proteoglycan (KSPG) from scar tissue of experimental penetrating corneal wounds in rabbits was analyzed 2-8 weeks after injury using three previously characterized antibodies. Keratan sulfate (KS) was identified in 2 week scars and normal corneal tissue by indirect immunofluorescence using a monoclonal antibody against sulfated KS epitopes. KSPG was measured in unfractionated extracts of scar and of normal corneal tissue using a "sandwich" enzyme-linked immunosorbent assay (ELISA). In extracts of 2 week scars, KSPG molecules reacting with two different anti-KS monoclonal antibodies were 55% and 82% as abundant as in normal tissue extracts. Ion exchange high performance liquid chromatography (HPLC) of tissue extracts found qualitatively similar elution profiles of KSPG antigens from both scar and normal tissues. Direct ELISA of the HPLC-purified KSPG showed identical quantitative binding of antibodies against core protein and KS from normal and scar tissue. KS in the HPLC-purified extracts was sensitive to digestion with endo-beta-galactosidase, whereas core protein antigens were not affected by this enzyme, as expected. Alteration of the antigenic characteristics of the KSPG of scars was detected with a competitive immunoassay using immobilized monoclonal antibodies against KS. KS in extracts from 2, 6, and 8 week scars competed only 5-11% as effectively as KS from normal cornea, although core protein antigens in the scar extracts competed 61-80% as well as those of normal cornea.(ABSTRACT TRUNCATED AT 250 WORDS)

67 citations


Journal ArticleDOI
Ronald L. Goldberg1
TL;DR: An enzyme-linked immunosorbent assay has been developed to measure hyaluronate concentrations in biological samples and can be used for measuring large numbers of biological samples from a variety of animal and tissue sources.

52 citations


Journal ArticleDOI
TL;DR: The results confirmed those recently presented by Yang et al that there may be subgroups of macular dystrophy that can be identified by immunohistochemical methods and identified at least three subgroups in the disease.
Abstract: • Macular corneal dystrophy is an autosomal recessive disorder in which abnormal deposits in the corneal stroma have been identified. We examined the corneal buttons of 12 patients, who had clinical features of macular dystrophy, by histochemical staining, transmission electron microscopy, and immunohistochemical techniques. All corneas exhibited positive staining with Muller Mowry's colloidal iron. Using monoclonal antibodies 1/20/ 5-D-4, J-10, J-19, and J-36 that recognize specific sites on the sulfated keratan sulfate molecule, we stained corneal sections by an avidin-biotin-peroxidase complex method and identified two groups of macular corneal dystrophy. One group consisting of four corneas reacted positively with all four antibodies, and the other group consisting of eight corneas did not react with any of the antibodies used. These results confirmed those recently presented by Yang et al that there may be subgroups of macular dystrophy that can be identified by immunohistochemical methods. Also, serum levels of sulfated keratan sulfate were determined in seven patients. One patient who displayed a normal level of serum keratan sulfate had positive corneal immunoreactivity. Of the six patients who lacked serum keratan sulfate, four showed negative and two had positive corneal immunostaining, suggesting at least three sub-groups in the disease. An attempt was made to correlate the clinical features, histochemical-staining characteristics, and ultrastructural morphology with the immunoreactivity to keratan sulfate antibodies, but no correlations could be made.

Journal ArticleDOI
TL;DR: It is suggested that male STR/IN mice are particularly useful for studying two different types of osteoarthrosis, one due to a biomechanically induced instability (patellar luxation) and oneDue to biochemical changes (absence of keratan sulfate) of still unknown pathogenesis.
Abstract: The development of osteoarthrotic cartilage lesions in the knee joints of male STR/IN mice was studied with respect to their histologic appearance and their various localizations in the joint. Spontaneous articular cartilage degeneration on the medial portion of the tibial plateau was considered to be the initial event. Continued loss of cartilage subsequently led to a pronounced instability of the knee joint, with a varus deformity. This was followed by medial patellar luxation with corresponding osteoarthrotic lesions at the facies patellaris femoris. The most marked osteoarthrotic cartilage degeneration developed on the medial tibial condyle and at the facies patellaris femoris of the femoropatellar joint. Histologic examination of the osteoarthrotic defects in these two regions revealed distinct morphologic differences with respect to formation of chondrocyte clusters, tendency to regeneration, and proliferation reactions. Lectin binding experiments in normal articular cartilage revealed regional differences regarding the presence or absence of keratan sulfate in the extracellular matrix. The lack of keratan sulfate in tibial cartilage might reflect its tendency to degenerate spontaneously. It is therefore suggested that male STR/IN mice are particularly useful for studying two different types of osteoarthrosis, one due to a biomechanically induced instability (patellar luxation) and one due to biochemical changes (absence of keratan sulfate) of still unknown pathogenesis.

Journal ArticleDOI
TL;DR: Quantitation of the glycosaminoglycans in both extracts revealed chondroitin sulfate to represent the major species present and hyaluronic acid was observed predominantly in the guanidine/EDTA extract while dermatan sulfate was a quantitative minor component of both extracts.
Abstract: The glycosaminoglycans in human cementum have been studied. Following proteolytic digestion of guanidine/EDTA and collagenase extracts of cementum, glycosaminoglycans were isolated and then separated by cellulose acetate membrane electrophoresis. After specific elimination by enzymatic and chemical treatments the glycosaminoglycans were identified as hyaluronic acid, chondroitin sulfate and dermatan sulfate. Neither heparan sulfate nor keratan sulfate were observed. Quantitation of the glycosaminoglycans in both extracts revealed chondroitin sulfate to represent the major species present. Hyaluronic acid was observed predominantly in the guanidine/EDTA extract while dermatan sulfate was a quantitative minor component of both extracts.

Journal ArticleDOI
TL;DR: It is suggested that the resistance of the disc KS to these digestive procedures arises from branching and/or sites of multisulfation on the polysaccharide chain andarose gel electrophoresis and compositional analyses of the keratan sulfates supported such an interpretation.

Journal ArticleDOI
TL;DR: Modulatory alterations in the knee articular cartilages of femoral and tibial medial condyles following 1-8 weeks of moderate running exercise of 4- to 6-month-old rabbits probably enhance the stability and elastic stiffness of the PG matrix.
Abstract: Proteoglycans (PGs) and collagen were quantified in the knee articular cartilages of femoral and tibial medial condyles following 1-8 weeks of moderate running exercise of 4- to 6-month-old rabbits. The total content of PGs extractable with 4 mol/l guanidium chloride was elevated in the weight-bearing cartilage of the tibial medial condyle, while their concentration, expressed as uronic acid per wet weight, and collagen remained unchanged. The content of glucosamine (GlcN) and its ratio to galactosamine (GalN) was elevated in femoral cartilage PGs purified by centrifugation in dissociative CsCl gradients, indicating an increase in keratan sulfate. After 8 weeks of running, the chondroitin sulfate chains of PGs from tibial medial condyle contained less unsulfated disaccharide units. The content of chondroitin sulfate was elevated in the nonextractable residue of the tibial medial condyle as indicated by uronic acid and GaN assays. The content of nonextractable GlcN was increased even more, both in tibial medial and femoral cartilages. Moderate running thus increased (1) keratan sulfate-rich PGs, (2) the degree of sulfation of the chondroitin sulfate chains, and (3) nonextractable PGs; these modulatory alterations probably enhance the stability and elastic stiffness of the PG matrix.

Journal ArticleDOI
TL;DR: Immunohistochemical localization utilizing monoclonal antibodies confirms the localization of chondroitin sulfate and keratan sulfate in the compressional regions and its absence in tensional areas, indicating that adult flexor tendon cells in culture continue to express their region-specific phenotypes.
Abstract: Rabbit flexor tendons have two distinct biomechanical regions: a compressional region which is characterized by chondrocyte-like cells and abundant matrix, and a tensional region which has a typical tendon morphology with elongated cells, sparse matrix and parallel bundles of collagen fibers. Tissue culture of these regions yields two distinct populations of cells. The compressional cells in vitro synthesize high molecular weight chondroitin sulfate proteoglycan, while the tensional cells synthesize a dermatan sulfate rich, low molecular weight proteoglycan. Immunohistochemical localization utilizing monoclonal antibodies confirms the localization of chondroitin sulfate and keratan sulfate in the compressional regions and its absence in tensional areas. These observations indicate that adult flexor tendon cells in culture continue to express their region-specific phenotypes.

Journal Article
TL;DR: Results show that elongation of both chondroitin sulfate and keratan sulfate glycosaminoglycans takes place in the same Golgi compartments, which include the middle cisternae and probably also the trans cisternAE and tubules.

Journal ArticleDOI
TL;DR: Fragments of large monomers containing keratan sulfate and hyaluronic acid binding region are the major component of similar fractions from old cartilage.
Abstract: Proteoglycans extracted from articular cartilage of large joints of humans aged 4, 11, 70 and 75, were fractionated on associative density gradients. The top fraction (A3) was purified by ion-exchange chromatography and subsequent gel filtration on Sepharose CL 4B in 4 M GuCl, 0.5% Triton x 100. Proteoglycans from young cartilages yielded a narrow rapid migrating band on gel electrophoresis, had a Kav of 0.43 and 0.44 on Sepharose CL 4B, a glucosamine/galactosamine ratio of 0.11 and 0.12 and a glycoprotein core rich in aspartic acid and leucine with a Mr of about 47,000. Proteoglycans from old cartilages gave a wider and slower migrating band on gel electrophoresis, had a wide peak with a Kav of 0.38 and 0.40 on Sepharose CL 4B, a glucosamine/galactosamine ratio of 5.1 and 3.2, a glycoprotein core rich in glutamic acid and glycine, and with a Mr of about 170,000-180,000. Analysis using monoclonal antibodies detected epitopes of keratarn sulfate and of hyaluronic acid binding region in the fractions from old but not in those from young cartilages. Small proteoglycans not derived from the large monomers are the major component of low-buoyant-density fractions of proteoglycans from young cartilages. Fragments of large monomers containing keratan sulfate and hyaluronic acid binding region are the major component of similar fractions from old cartilage.

Journal ArticleDOI
TL;DR: Proteoglycans were isolated from leptocephalous larvae of the bonefish, which were in the early stages of metamorphosis, using both associative and dissociative conditions in the presence of protease inhibitors to achieve an extraction efficiency of 75% and 85–90% respectively.
Abstract: Proteoglycans (PGs) were isolated from leptocephalous larvae of the bonefish (Albula sp.), which were in the early stages of metamorphosis, using both associative and dissociative conditions in the presence of protease inhibitors. The procedure was rapid and resulted in an extraction efficiency of 75% (associative) and 85-90% (dissociative). The majority of co-extracted protein could be effectively separated from the PGs by utilizing either Sepharose CL-2B or CL-6B gel chromatography. Sepharose CL-2B chromatography of extracted PGs after treatment with bacterial keratan sulfate-endo-β-galactosidase (keratanase) showed that most of the high molecular weight (M r) carbohydrate was degraded. Free keratan sulfate (KS) chains were prepared from whole-larva extracts (which also contain small amounts of chondroitin sulfate) by both chondroitinase ABC treatment and ethanol fractionation. Sepharose CL-6B chromatography under dissociative conditions showed that larval KS chains were much larger (M r∼55,000) than those from cornea. These chains tended to aggregate when chromatographed under associative conditions. Larval KS was degraded by keratanase and resistant to chondroitinase, ABC and testicular hyaluronidase. Differences were also noted in the oligosaccharides produced by keratanase treatment of the two preparations. However, biochemical composition of larval and corneal KS was similar.

Journal ArticleDOI
TL;DR: It is indicated that chondroitin sulfate and keratan sulfate chains are both present in the proteoglycan extract, and within limits of detection, all core proteins belong to the high-molecular-weight category.
Abstract: Monoclonal antibodies directed against specific carbohydrate epitopes on chondroitin 4-/dermatan sulfate, chondroitin 6-sulfate, keratan sulfate, and a monoclonal antibody directed against the hyaluronate binding region were used to characterize proteoglycans extracted from embryonic chick bone marrow. About half of the proteoglycans separate into the high density fraction on a CsCl gradient. Glycosaminoglycan-specific antibodies recognize proteoglycans from all fractions; this includes an antibody directed against keratan sulfate. Some proteoglycans, principally in the high buoyant density fraction, contain sites recognized by the antibody specific for the hyaluronate binding region. Within limits of detection, all core proteins belong to the high-molecular-weight category, with weights in excess of 212 kD. Antibodies directed against chondroitin 4-/dermatan sulfate and against keratan sulfate primarily bind to extracellular matrix material located in the extracellular spaces and to matrix elements in the pericellular regions of fibroblastic stromal cells. The antibody that recognizes chondroitin 6-sulfate binds to sites on surfaces of fibroblastic stromal cells and also to extracellular matrix material. Little or no antibody binding is detected on surfaces of granulocytic cells. These studies indicate that chondroitin sulfate and keratan sulfate chains are both present in the proteoglycan extract.

Journal ArticleDOI
TL;DR: The results of this study showed that this new sensitive approach, which requires as little as 200 μg wet cartilage as starting material, provides important qualitative and quantitative information about the major constituents of the matrix.
Abstract: Very thin slices of the superficial and deep layers of osteophytic and apparently normal articular cartilage from six human osteoarthritic femoral heads were digested with papain. The digests were analyzed for keratan sulfate content using an enzyme-linked immunosorbent assay (ELISA) with inhibition step, and for chondroitin sulfate and collagen contents using biochemical assays. Although there were marked differences in Safranin-O staining of the superficial and deep layers of osteophytic cartilage, these two layers had identical high ratios of chondroitin sulfate/collagen. Keratan sulfate was present only in small amounts in osteophytic cartilage. However, the deeper layer contained significantly more of this glycosaminoglycan. The deeper layer of articular cartilage contained approximately twice as much chondroitin sulfate and six times more keratan sulfate relative to collagen than the superficial layer. The results of this study showed that this new sensitive approach, which requires as little as 200 μg wet cartilage as starting material, provides important qualitative and quantitative information about the major constituents of the matrix.

Journal ArticleDOI
TL;DR: Rabbit meniscal fibrochondrocytes grown in vitro under culture conditions previously shown to foster growth of this cell type synthesized sulfated proteoglycans which could be differentiated by their solubility when dialyzed against water, demonstrating their differentiated phenotype in short-term monolayer cell culture.

Journal ArticleDOI
TL;DR: Human serum amyloid P component was found to agglutinate erythrocytes in the presence of calcium ion and the hemagglutination was strongly inhibited by hyaluronic acid as well as by heparan sulfate and dermatan sulfates, but not by chondroitin 4-sulfate and keratan sulfate.

Journal Article
TL;DR: The contribution made by notochordal-derived keratan sulphate to the glycosaminoglycan content of the mature intervertebral disc may differ in man from that of other animal species.
Abstract: Intervertebral discs, formed from notochord cell expansions during embryogenesis, are known to contain proteoglycans bearing keratan sulphate chains. Keratan sulphate has previously been demonstrated in Xenopus and chick notochords and in human fetal cartilage. In contrast, we have been unable to demonstrate keratan sulphate in human fetal notochord using two monoclonal antibodies, MZ15 and 5-D-4. The contribution made by notochordal-derived keratan sulphate to the glycosaminoglycan content of the mature intervertebral disc may differ in man from that of other animal species.

Journal ArticleDOI
01 Jun 1988-Spine
TL;DR: Measurements of blood levels of keratan sulfate could prove useful in monitoring effective degradation of disc proteoglycans in chemonucleolysis in man and help discriminate between ineffective enzyme placement, and alternative mechanisms of treatment failure.
Abstract: Chymopapain (1 mg) was injected into each of four-lumbar intervertebral discs of adult mongrel dogs. As expected, at 2 weeks, all injected discs exhibited marked loss of height (mean: 50% of original height) indicative of severe proteoglycan depletion. The appearance of keratan sulfate-bearing fragments in plasma was monitored by an ELISA-inhibition assay which uses a monoclonal antibody (1/20/5-D-4) specific for an epitope present only in the longest keratan sulfate chains. Levels of plasma keratan sulfate rose within 30 minutes and reached a maximum between 24 and 72 hours later. Levels then declined progressively but were still elevated at 2 weeks postinjection. Keratan sulfate-bearing fragments in plasma were purified by ion exchange chromatography on DEAE-Sephacryl and fractionated by sieve chromatography on Sepharose CL-6B. These plasma keratan sulfate-bearing fragments were found to be similar in size to keratan sulfate-bearing fragments generated by chymopapain digestion of dog nucleus pulposus proteoglycans, but slightly larger than single keratan sulfate chains obtained by alkaline borohydride treatment of dog nucleus pulposus proteoglycans. The results of this study show that measurements of blood levels of keratan sulfate could prove useful in monitoring effective degradation of disc proteoglycans in chemonucleolysis in man and help discriminate between ineffective enzyme placement, and alternative mechanisms of treatment failure.

Journal ArticleDOI
TL;DR: The total GAG accumulation in the medium was much greater than that of the cell layer indicating that the cells are synthesizing relatively large amounts of GAGs, although incorporation of these macromolecules into the extracellular matrix was consistently low.

Journal ArticleDOI
TL;DR: It was suggested that heparin potentiates the lytic activity of perforin and acid mucopolysaccharides may actually be involved in target cell lysis by CTL.
Abstract: A cytolytic protein (perforin) was rapidly purified from a cell line of mouse cytotoxic T-lymphocytes (CTL) by DEAE-cellulose, heparin-Sepharose, and phenyl-Sepharose chromatographies. The purified perforin was activated by heparin, the half maximal concentration being 3-10 ng/ml, depending on the calcium concentration. Other acid mucopolysaccharides, such as chondroitin sulfates A and C, keratan polysulfate, and heparin sulfate, also enhanced the lysis of erythrocytes by perforin, but the concentrations required for activation were more than 100-fold higher than that of heparin. Chondroitin, hyaluronic acid, and keratan sulfate, however, had no effect on the perforin activity. It was suggested that heparin potentiates the lytic activity of perforin and acid mucopolysaccharides may actually be involved in target cell lysis by CTL.

Journal ArticleDOI
01 Jan 1988-Bone
TL;DR: In an attempt to elucidate the biochemical defect in pseudoachondroplasia, proteoglycan metabolism was investigated in cartilage from a patient with the dominant form of this condition and no differences between the proteoglycans of pseudoachONDroplastic and normal cartilage were detected.

DOI
01 Jan 1988
TL;DR: Use of a monoclonal antibody to the glycosaminoglycan moiety of a keratan sulfate proteoglycan demonstrated (at the light microscopic level) that CA contains antigenic sites for keratan oxide proteoglycans.
Abstract: Corpora amylacea (CA) are spherical bodies, varying in diameter (10–100 microns), which accumulate in the brain during normal aging and in a number of diseases. At the ultrastructural level CA contain masses of randomly oriented short linear fibrillar structures (14). In many instances the complete CA granule is surrounded by glial filaments. The presence of CA in the brain has been recognized for over a century, however its definite composition and source of origin is relatively unknown. CA tend to accumulate in subependymal, subpial and perivascular regions and are very prominent in the olfactory tract during normal senescence. Early investigations into the nature of CA suggested the presence of glucose polymers, maltose (15) and other types of carbohydrates (1), with small amounts of protein (15). A more recent study using various lectins to stain CA suggested the presence of D-glucose, D-mannose, D-galactose, alpha-L-fucose and N-acetylgalactosamine (12). In addition use of a monoclonal antibody to the glycosaminoglycan moiety of a keratan sulfate proteoglycan demonstrated (at the light microscopic level) that CA contains antigenic sites for keratan sulfate proteoglycans (12).

Book ChapterDOI
01 Jan 1988
TL;DR: The basic dye: 1,9-dimethylmethylene blue (DMB) is one of the most sensitive thiazine compounds, giving a more marked metachromatic shift than other thiazines when mixed with glycosaminoglycans (GAGs).
Abstract: The basic dye: 1,9-dimethylmethylene blue (DMB) is one of the most sensitive thiazine compounds, giving a more marked metachromatic shift than other thiazine dyes when mixed with glycosaminoglycans (GAGs).