Biosynthesis of promatrix metalloproteinase-9/chondroitin sulphate proteoglycan heteromer involves a Rottlerin-sensitive pathway.

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Nabin Malla

Full Text Available BACKGROUND: Previously we have shown that a fraction of the matrix metalloproteinase-9 (MMP-9 synthesized by the macrophage cell line THP-1 was bound to a chondroitin sulphate proteoglycan (CSPG core protein as a reduction sensitive heteromer. Several biochemical properties of the enzyme were changed when it was bound to the CSPG. METHODOLOGY/PRINCIPAL FINDINGS: By use of affinity chromatography, zymography, and radioactive labelling, various macrophage stimulators were tested for their effect on the synthesis of the proMMP-9/CSPG heteromer and its components by THP-1 cells. Of the stimulators, only PMA largely increased the biosynthesis of the heteromer. As PMA is an activator of PKC, we determined which PKC isoenzymes were expressed by performing RT-PCR and Western Blotting. Subsequently specific inhibitors were used to investigate their involvement in the biosynthesis of the heteromer. Of the inhibitors, only Rottlerin repressed the biosynthesis of proMMP-9/CSPG and its two components. Much lower concentrations of Rottlerin were needed to reduce the amount of CSPG than what was needed to repress the synthesis of the heteromer and MMP-9. Furthermore, Rottlerin caused a minor reduction in the activation of the PKC isoenzymes δ, ε, θ and υ (PKD3 in both control and PMA exposed cells. CONCLUSIONS/SIGNIFICANCE: The biosynthesis of the proMMP-9/CSPG heteromer and proMMP-9 in THP-1 cells involves a Rottlerin-sensitive pathway that is different from the Rottlerin sensitive pathway involved in the CSPG biosynthesis. MMP-9 and CSPGs are known to be involved in various physiological and pathological processes. Formation of complexes may influence both the specificity and localization of the enzyme. Therefore, knowledge about biosynthetic pathways and factors involved in the formation of the MMP-9/CSPG heteromer may contribute to insight in the heteromers biological function as well as pointing to future targets for therapeutic agents.

Basement membrane proteoglycans in glomerular morphogenesis: chondroitin sulfate proteoglycan is temporally and spatially restricted during development

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McCarthy, K J; Bynum, K; St John, P L

1993-01-01

We previously reported the presence of a basement membrane-specific chondroitin sulfate proteoglycan (BM-CSPG) in basement membranes of almost all adult tissues. However, an exception to this ubiquitous distribution was found in the kidney, where BM-CSPG was absent from the glomerular capillary......, the present study used light and electron microscopic immunohistochemistry to examine the distribution of BM-CSPG and basement membrane heparan sulfate proteoglycan (BM-HSPG) during prenatal and postnatal renal development in the rat. Our results show that the temporal and spatial pattern of expression of BM...

Basement membrane chondroitin sulfate proteoglycan alterations in a rat model of polycystic kidney disease

DEFF Research Database (Denmark)

Ehara, T; Carone, F A; McCarthy, K J

1994-01-01

of distal tubules and collecting ducts was observed by 4 days with phenol II treatment, but the morphology returned to normal after 7 days of subsequent normal diet. Staining of tissue sections with two mouse monoclonal antibodies to a recently described basement membrane chondroitin sulfate proteoglycan...... to chondroitin sulfate chains confirmed these changes in cystic tubule basement membranes. During the recovery stage, interstitial chondroitin sulfate (representing a CSPG other than BM-CSPG) was greatly increased around these tubules, along with the glycoprotein fibronectin. Staining with antibody to a basement...... membrane heparan sulfate proteoglycan core protein related to perlecan did not diminish but rather stained affected tubules intensely, whereas laminin, on the other hand, was apparently diminished in the basement membranes of the cystic tubules. Type IV collagen staining did not change through disease...

Molecular characterization and transcriptional analysis of the female-enriched chondroitin proteoglycan 2 of Toxocara canis.

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Ma, G X; Zhou, R Q; Hu, L; Luo, Y L; Luo, Y F; Zhu, H H

2018-03-01

Toxocara canis is an important but neglected zoonotic parasite, and is the causative agent of human toxocariasis. Chondroitin proteoglycans are biological macromolecules, widely distributed in extracellular matrices, with a great diversity of functions in mammals. However, there is limited information regarding chondroitin proteoglycans in nematode parasites. In the present study, a female-enriched chondroitin proteoglycan 2 gene of T. canis (Tc-cpg-2) was cloned and characterized. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to measure the transcription levels of Tc-cpg-2 among tissues of male and female adult worms. A 485-amino-acid (aa) polypeptide was predicted from a continuous 1458-nuleotide open reading frame and designated as TcCPG2, which contains a 21-aa signal peptide. Conserved domain searching indicated three chitin-binding peritrophin-A (CBM_14) domains in the amino acid sequence of TcCPG2. Multiple alignment with the inferred amino acid sequences of Caenorhabditis elegans and Ascaris suum showed that CBM_14 domains were well conserved among these species. Phylogenetic analysis suggested that TcCPG2 was closely related to the sequence of chondroitin proteoglycan 2 of A. suum. Interestingly, a high level of Tc-cpg-2 was detected in female germline tissues, particularly in the oviduct, suggesting potential roles of this gene in reproduction (e.g. oogenesis and embryogenesis) of adult T. canis. The functional roles of Tc-cpg-2 in reproduction and development in this parasite and related parasitic nematodes warrant further functional studies.

Modulation of the expression of chondroitin sulfate proteoglycan in stimulated human monocytes

International Nuclear Information System (INIS)

Uhlin-Hansen, L.; Eskeland, T.; Kolset, S.O.

1989-01-01

Proteoglycan biosynthesis was studied in human monocytes and monocyte-derived macrophages (MDM) after exposure to typical activators of the monocyte/macrophage system: interferon-gamma (IFN-gamma), lipopolysaccharide (LPS), and phorbol 12-myristate 13-acetate (PMA). By morphological examination, both monocytes and MDM were stimulated by these activators. Treatment with IFN-gamma resulted in a slight decrease in the expression of [35S]chondroitin sulfate proteoglycan (CSPG) in both monocytes and MDM, whereas LPS treatment increased the [35S]CSPG expression 1.8 and 2.2 times, respectively. PMA, in contrast, decreased the CSPG expression 0.4 times in monocytes, whereas MDM were stimulated to increase the biosynthesis 1.9 times. An increase in the sulfate density of the chondroitin sulfate chains was evident following differentiation of monocytes into MDM due to the expression of disulfated disaccharide units of the chondroitin sulfate E type (CS-E). However, monocytes exposed to PMA did also express disaccharides of the chondroitin sulfate E type. Furthermore, the expression of CS-E in MDM was increased 2 times following PMA treatment. An inactive phorbol ester, phorbol 12,13-diacetate, did not affect the expression of CS-E in either monocytes or MDM when compared with control cultures, suggesting that protein kinase C-dependent signal pathways may be involved in the regulation of sulfation of CSPG. Exposure to LPS or IFN-gamma did not lead to any changes in the sulfation of the chondroitin sulfate chains

Expression of the chondroitin sulphate proteoglycan molecular complex in six human melanoma xenograft lines studied by flow cytometry and immunohistochemistry.

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Nagelhus, T A; Rofstad, E K

1993-06-01

The expression of the chondroitin sulphate proteoglycan (CSP) molecular complex in six human melanoma xenograft lines (BEX-t, COX-t, HUX-t, ROX-t, SAX-t, WIX-t) was studied by flow cytometry and immunohistochemistry using the monoclonal antibodies 9.2.27, ME31.3, G7A5, and NKI.M6. The two methods and the four antibodies gave consistent results. The six melanoma lines could be divided into three distinct groups of two lines each; expression was high in the HUX-t and ROX-t lines and intermediate in the BEX-t and SAX-t lines, whereas the COX-t and WIX-t lines were negative. The mean number of epitopes per cell for 9.2.27 was approximately twice as high as for ME31.3, G7A5, and NKI.M6 and was estimated to range from 0.8 +/- 0.1 x 10(5) to 1.9 +/- 0.2 x 10(5) in the positive xenograft lines. The expression of the CSP complex was heterogeneous. The immunofluorescence histograms measured by flow cytometry were therefore broad for all tumour lines. A significant fraction of the HUX-t cells was negative or weakly stained. These cells appeared as clear negative patches in the immunohistochemical preparations. Moreover, most morphologically intact tumour cells adjacent to necrotic areas did not show significant expression of the CSP complex, irrespective of tumour line. These cells were probably hypoxic and thus resistant to radiation therapy. The expression of the CSP complex in the xenograft lines was similar to that reported for melanoma in man.

Ultrastructural immunocytochemical localization of chondroitin sulfate proteoglycan in Bruch's membrane of the rat

DEFF Research Database (Denmark)

Lin, W L; Essner, E; McCarthy, K J

1992-01-01

Two monoclonal antibodies (Mab 4D5 and 2D6) raised against the core protein of a basement membrane chondroitin sulfate proteoglycan from Reichert's membrane of the rat, were used for ultrastructural immunoperoxidase localization of this protein in Bruch's membrane of the rat. Immunoreactivity...

Human recombinant Fab fragment from combinatorial libraries of a B-cell lymphoma patient recognizes core protein of chondroitin sulphate proteoglycan 4.

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Egami, Yoko; Narushima, Yuta; Ohshima, Motohiro; Yoshida, Akira; Yoneta, Naruki; Masaki, Yasufumi; Itoh, Kunihiko

2018-01-01

CD antigens are well known as therapeutic targets of B-cell lymphoma. To isolate therapeutic antibodies that recognize novel targets other than CD antigens, we constructed a phage display combinatorial antibody Fab library from bone marrow lymphocytes of B-cell lymphoma patient. To eliminate antibodies reactive with known B-cell lymphoma antigen, non-hematopoietic and patient's sera reactive HeLaS3 cells was selected as a target of whole cell panning. Five rounds of panning against live HeLaS3 cells retrieved single Fab clone, termed AHSA (Antibody to HeLa Surface Antigen). Using phage display random peptide library, LSYLEP was identified as an epitope sequence of AHSA. LC-MS/MS analysis of AHSA-precipitated HeLaS3 cell lysates detected several fragments corresponding to the sequence of chondroitin sulphate proteoglycan 4 (CSPG4) core protein. Since LSYLEP sequence was at the position of 313-318 of CSPG4, we considered that CSPG4 was AHSA-associated antigen. Double staining of CSPG4-postive MDA-MB-435S cells with AHSA and anti-CSPG4 rabbit antibody showed identical staining position, and reduced AHSA reactivity was observed in CSPG4-siRNA treated MDA-MB-435S cells. In conclusion, we retrieved a human Fab from antibody library of B-cell lymphoma patient, and identified CSPG4 as a recognizing antigen. AHSA may have potential benefits for development of CSPG4-targeting theranostics for B-cell lymphoma. © The Authors 2017. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

SRPX2 is a novel chondroitin sulfate proteoglycan that is overexpressed in gastrointestinal cancer.

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Kaoru Tanaka

Full Text Available SRPX2 (Sushi repeat-containing protein, X-linked 2 has recently emerged as a multifunctional protein that is involved in seizure disorders, angiogenesis and cellular adhesion. Here, we analyzed this protein biochemically. SRPX2 protein was secreted with a highly posttranslational modification. Chondroitinase ABC treatment completely decreased the molecular mass of purified SRPX2 protein to its predicted size, whereas heparitinase, keratanase and hyaluroinidase did not. Secreted SRPX2 protein was also detected using an anti-chondroitin sulfate antibody. These results indicate that SRPX2 is a novel chondroitin sulfate proteoglycan (CSPG. Furthermore, a binding assay revealed that hepatocyte growth factor dose-dependently binds to SRPX2 protein, and a ligand-glycosaminoglycans interaction was speculated to be likely in proteoglycans. Regarding its molecular architecture, SRPX2 has sushi repeat modules similar to four other CSPGs/lecticans; however, the molecular architecture of SRPX2 seems to be quite different from that of the lecticans. Taken together, we found that SRPX2 is a novel CSPG that is overexpressed in gastrointestinal cancer cells. Our findings provide key glycobiological insight into SRPX2 in cancer cells and demonstrate that SRPX2 is a new member of the cancer-related proteoglycan family.

Recent Insights into Cell Surface Heparan Sulphate Proteoglycans and Cancer

DEFF Research Database (Denmark)

Couchman, John R; Multhaupt, Hinke; Sanderson, Ralph D

2016-01-01

behaviour. Here, we review some recent advances, emphasising that many tumour-related functions of proteoglycans are revealed only after their modification in processes subsequent to synthesis and export to the cell surface. These include enzymes that modify heparan sulphate structure, recycling of whole...

Developmental and functional significance of the CSF-1 proteoglycan chondroitin sulfate chain

OpenAIRE

Nandi, Sayan; Akhter, Mohammed P.; Seifert, Mark F.; Dai, Xu-Ming; Stanley, E. Richard

2006-01-01

The primary macrophage growth factor, colony-stimulating factor-1 (CSF-1), is homodimeric and exists in 3 biologically active isoforms: a membrane-spanning, cell-surface glycoprotein (csCSF-1) and secreted glycoprotein (sgCSF-1) and proteoglycan (spCSF-1) isoforms. To investigate the in vivo role of the chondroitin sulfate glycosaminoglycan (GAG) chain of spCSF-1, we created mice that exclusively express, in a normal tissue-specific and developmental manner, either the secreted precursor of s...

Intravesical Glycosaminoglycan Replacement with Chondroitin Sulphate (Gepan(®) instill) in Patients with Chronic Radiotherapy- or Chemotherapy-Associated Cystitis.

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Schwalenberg, Thilo; Berger, Frank Peter; Horn, Lars Christian; Thi, Phuc Ho; Stolzenburg, Jens-Uwe; Neuhaus, Jochen

2015-08-01

Intravesical instillation of glycosaminoglycans is a promising option for the treatment of chronic cystitis, as it supports the regeneration of the damaged urothelial layer. We investigated the efficacy of short-term intravesical chondroitin sulphate treatment (six courses of instillation) in patients with chronic radiotherapy- or chemotherapy-associated cystitis. This prospective, observational study included patients with chronic radiotherapy- or chemotherapy-associated cystitis, who received six once-weekly intravesical instillations of 0.2% chondroitin sulphate 40 mL. Every week, patients recorded their symptoms and their benefits and tolerance of treatment, using a self-completed questionnaire. The study included 16 patients (mean age 68.5 years; 50% male). During the study, a reduction in all evaluated parameters was observed. After one dose of chondroitin sulphate, symptom improvement was observed in 38% of patients, and after the second dose, an additional 31% of patients showed improvement. At week 6, 80% of patients had either improved or were symptom free, and significant improvements in urinary urgency (p = 0.0082), pollakisuria (p = 0.0022), urge frequency (p = 0.0033) and lower abdominal pain (p = 0.0449) were observed. Haematuria, present in 9 of the 16 patients at baseline, was completely resolved in all cases after 6 weeks. The majority of patients (93%) evaluated the tolerance of chondroitin sulphate as 'good' or 'very good'. No treatment-related adverse events were reported. Intravesical administration of chondroitin sulphate was effective for the treatment of radiotherapy- or chemotherapy-associated cystitis. Even short-term treatment appears to be effective in reducing symptoms and improving the quality of life of patients.

Mapping by monoclonal antibody detection of glycosaminoglycans in connective tissues

DEFF Research Database (Denmark)

Couchman, J R; Caterson, B; Christner, J E

1984-01-01

Chondroitin sulphate proteoglycans are widespread connective tissue components and chemical analysis of cartilage and other proteoglycans has demonstrated molecular speciation involving the degree and position of sulphation of the carbohydrate chains. This may, in turn, affect the properties...... of the glycosaminoglycan (GAG), particularly with respect to self-association and interactions with other extracellular matrix components. Interactions with specific molecules from different connective tissue types, such as the collagens and their associated glycoproteins, could be favoured by particular charge...... and dermatan sulphate. These provide novel opportunities to study the in vivo distribution of chondroitin sulphate proteoglycans. We demonstrate that chondroitin sulphates exhibit remarkable connective tissue specificity and furthermore provide evidence that some proteoglycans may predominantly carry only one...

Hair follicle proteoglycans

DEFF Research Database (Denmark)

Couchman, J R

1993-01-01

that are present in the epithelial and stromal compartments of hair follicles. However, the transmembrane proteoglycan syndecan may be important in follicle morphogenesis, both with respect to the epithelium and dermal papilla cells. Syndecan may possess both heparan and chondroitin sulfate chains, interacts...... basement membranes, including those surrounding the epithelial compartment of hair follicles. Additionally, and quite unlike the dermis, the dermal papilla is enriched in basement-membrane components, especially a chondroitin 6-sulfate-containing proteoglycan, BM-CSPG. The function of this proteoglycan...... is not known, but developmental studies indicate that it may have a role in stabilizing basement membranes. In the hair cycle, BM-CSPG decreases through catagen and is virtually absent from the telogen papilla. One or more heparan sulfate proteoglycans, including perlecan, are also present in papilla...

Differentiation of 3T3-L1 preadipocytes with 3-isobutyl-1-methylxanthine and dexamethasone stimulates cell-associated and soluble chondroitin 4-sulfate proteoglycans

International Nuclear Information System (INIS)

Calvo, J.C.; Rodbard, D.; Katki, A.; Chernick, S.; Yanagishita, M.

1991-01-01

The proteoglycans (cell-associated and culture media) in 3T3-L1 preadipocytes in culture were analyzed before and during differentiation into adipocytes. Cells were metabolically labeled with [35S]sulfate and [3H] glucosamine for 24 h and then extracted and analyzed. There was a 1.68 ± 0.07-fold increase in the 35S in medium proteoglycan during differentiation, whereas cell-associated proteoglycan radioactivity showed no increase. Analyses of radiolabeled molecules using ion-exchange chromatography, gel filtration, and high performance liquid chromatography after enzymatic or alkaline digestion indicated that all of the 35S label was recovered as two major species of chondroitin 4-sulfate proteoglycans (CSPG-I and CSPG-II) and 7% as heparan sulfate proteoglycan. CSPG-I has a mass of ∼ 970 kDa with multiple chondroitin sulfate chains (average of 50 kDa each) and a core protein of ∼ 370 kDa including oligosaccharides. CSPG-II has a mass of 140 kDa with one or two chondroitin sulfate chains (average of 68 kDa each) and a core protein of 41 kDa including oligosaccharides. CSPG-I appears to be similar to versican, whereas CSPG-II is similar to decorin and/or biglycan, found in other fibroblastic cells. Cell differentiation was associated with a specific increase in CSPG-I (4.0 ± 0.2-fold in media and 3.2 ± 0.5-fold in the cell-associated form). This system should facilitate study of the functional roles of proteoglycans during growth and differentiation

Pattern of chondroitin sulfate proteoglycan expression after ablation of the sensorimotor cortex of the neonatal and adult rat brain

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Dacić Sanja

2008-01-01

Full Text Available The central nervous system has a limited capacity for self-repair after damage. However, the neonatal brain has agreater capacity for recovery than the adult brain. These differences in the regenerative capability depend on local environmental factors and the maturational stage of growing axons. Among molecules which have both growth-promoting and growth-inhibiting activities is the heterogeneous class of chondroitin sulfate proteoglycans (CSPGs. In this paper, we investigated the chondroitin-4 and chondroitin-6 sulfate proteoglycan expression profile after left sensorimotor cortex ablation of the neonatal and adult rat brain. Immunohistochemical analysis revealed that compared to the normal uninjured cortex, lesion provoked up regulation of CSPGs showing a different pattern of expression in the neonatal vs. the adult brain. Punctuate and membrane-bound labeling was predominate after neonatal lesion, where as heavy deposition of staining in the extracellular matrix was observed after adult lesion. Heavy deposition of CSPG immunoreactivity around the lesionsite in adult rats, in contrast to a less CSPG-rich environment in neonatal rats, indicated that enhancement of the recovery process after neonatal injury is due to amore permissive environment.

Levels of Urinary Trypsin Inhibitor and Structure of Its Chondroitin Sulphate Moiety in Type 1 and Type 2 Diabetes

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Ucciferri, Nadia; Idini, Michela; De Muro, Pierina

2018-01-01

Background Diabetes mellitus is a global health problem representing the fifth leading cause of mortality and a major risk factor for cardiovascular diseases. In the last years, we reported an association among urinary trypsin inhibitor (UTI), a small proteoglycan that plays pleiotropic roles in many inflammatory processes, and both type 1 and 2 diabetes and developed a method for its direct quantitation and structural characterization. Methods Urine from 39 patients affected by type 1 diabetes, 32 patients with type 2 diabetes, and 52 controls were analysed. UTI was separated from the main glycosaminoglycans physiologically present in urine by anion exchange chromatography, treated for chondroitin sulphate (CS) chain complete depolymerisation, and analysed for both UTI content and CS structure. UTI identification was performed by nano-LC-MS/MS analysis. Results We evidenced increased UTI levels, as well as reduced sulphation of its CS moiety in association with diabetes, regardless of both age and medium-term glycaemic control. Furthermore, no association between UTI and albumin excretion rate was found. Conclusions Evidences suggest that UTI levels are not directly correlated with renal function or, otherwise, that they may increase before the onset of renal impairment in diabetes, representing a potential marker for the underlying inflammatory condition. PMID:29541644

Transforming growth factor (type beta) promotes the addition of chondroitin sulfate chains to the cell surface proteoglycan (syndecan) of mouse mammary epithelia

OpenAIRE

1989-01-01

Cultured monolayers of NMuMG mouse mammary epithelial cells have augmented amounts of cell surface chondroitin sulfate glycosaminoglycan (GAG) when cultured in transforming growth factor-beta (TGF-beta), presumably because of increased synthesis on their cell surface proteoglycan (named syndecan), previously shown to contain chondroitin sulfate and heparan sulfate GAG. This increase occurs throughout the monolayer as shown using soluble thrombospondin as a binding probe. However, comparison o...

EGFR Activation Mediates Inhibition of Axon Regeneration by Myelin and Chondroitin Sulfate Proteoglycans

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Koprivica, Vuk; Cho, Kin-Sang; Park, Jong Bae; Yiu, Glenn; Atwal, Jasvinder; Gore, Bryan; Kim, Jieun A.; Lin, Estelle; Tessier-Lavigne, Marc; Chen, Dong Feng; He, Zhigang

2005-10-01

Inhibitory molecules associated with myelin and the glial scar limit axon regeneration in the adult central nervous system (CNS), but the underlying signaling mechanisms of regeneration inhibition are not fully understood. Here, we show that suppressing the kinase function of the epidermal growth factor receptor (EGFR) blocks the activities of both myelin inhibitors and chondroitin sulfate proteoglycans in inhibiting neurite outgrowth. In addition, regeneration inhibitors trigger the phosphorylation of EGFR in a calcium-dependent manner. Local administration of EGFR inhibitors promotes significant regeneration of injured optic nerve fibers, pointing to a promising therapeutic avenue for enhancing axon regeneration after CNS injury.

cDNA cloning of the basement membrane chondroitin sulfate proteoglycan core protein, bamacan: a five domain structure including coiled-coil motifs

DEFF Research Database (Denmark)

Wu, R R; Couchman, J R

1997-01-01

Basement membranes contain several proteoglycans, and those bearing heparan sulfate glycosaminoglycans such as perlecan and agrin usually predominate. Most mammalian basement membranes also contain chondroitin sulfate, and a core protein, bamacan, has been partially characterized. We have now....... The protein sequence has low overall homology, apart from very small NH2- and COOH-terminal motifs. At the junctions between the distal globular domains and the coiled-coil regions lie glycosylation sites, with up to three N-linked oligosaccharides and probably three chondroitin chains. Three other Ser...

Structural characterization of proteoglycans produced by testicular peritubular cells and Sertoli cells

International Nuclear Information System (INIS)

Skinner, M.K.; Fritz, I.B.

1985-01-01

The structural characteristics of proteoglycans produced by seminiferous peritubular cells and by Sertoli cells are defined. Peritubular cells secrete two proteoglycans designated PC I and PC II. PC I is a high molecular mass protein containing chondroitin glycosaminoglycan (GAG) chains (maximum 70 kDa). PC II has a protein core of 45 kDa and also contains chondroitin GAG chains (maximum 70 kDa). Preliminary results imply that PC II may be a degraded or processed form of PC I. Sertoli cells secrete two different proteoglycans, designated SC I and SC II. SC I is a large protein containing both chondroitin (maximum 62 kDa) and heparin (maximum 15 kDa) GAG chains. Results obtained suggest that this novel proteoglycan contains both chondroitin and heparin GAG chains bound to the same core protein. SC II has a 50-kDa protein core and contains chondroitin (maximum 25 kDa) GAG chains. A proteoglycan obtained from extracts of Sertoli cells is described which contains heparin (maximum 48 kDa) GAG chains. In addition, Sertoli cells secrete a sulfoprotein, SC III, which is not a proteoglycan. The stimulation by follicle-stimulating hormone of the incorporation of [ 35 S]SO 2 ) -4 ) into moieties secreted by Sertoli cells is shown to represent an increased production or sulfation of SC III, and not an increased production or sulfation of proteoglycans. Results are discussed in relation to the possible functions of proteoglycans in the seminiferous tubule

Large-scale chondroitin sulfate proteoglycan digestion with chondroitinase gene therapy leads to reduced pathology and modulates macrophage phenotype following spinal cord contusion injury

NARCIS (Netherlands)

Bartus, Katalin; James, Nicholas D; Didangelos, Athanasios; Bosch, Karen D; Verhaagen, J.; Yáñez-Muñoz, Rafael J; Rogers, John H; Schneider, Bernard L; Muir, Elizabeth M; Bradbury, Elizabeth J

2014-01-01

Chondroitin sulfate proteoglycans (CSPGs) inhibit repair following spinal cord injury. Here we use mammalian-compatible engineered chondroitinase ABC (ChABC) delivered via lentiviral vector (LV-ChABC) to explore the consequences of large-scale CSPG digestion for spinal cord repair. We demonstrate

Site-specific identification of heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans.

Science.gov (United States)

Noborn, Fredrik; Gomez Toledo, Alejandro; Green, Anders; Nasir, Waqas; Sihlbom, Carina; Nilsson, Jonas; Larson, Göran

2016-10-03

Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. The polysaccharides often display opposite effects on cell functions with HS and CS structural motifs presenting unique binding sites for specific ligands. Still, the mechanisms by which glycan biosynthesis generates complex HS and CS polysaccharides required for the regulation of mammalian physiology remain elusive. Here we present a glycoproteomic approach that identifies and differentiates between HS and CS attachment sites and provides identity to the core proteins. Glycopeptides were prepared from perlecan, a complex proteoglycan known to be substituted with both HS and CS chains, further digested with heparinase or chondroitinase ABC to reduce the HS and CS chain lengths respectively, and thereafter analyzed by nLC-MS/MS. This protocol enabled the identification of three consensus HS sites and one hybrid site, carrying either a HS or a CS chain. Inspection of the amino acid sequence at the hybrid attachment locus indicates that certain peptide motifs may encode for the chain type selection process. This analytical approach will become useful when addressing fundamental questions in basic biology specifically in elucidating the functional roles of site-specific glycosylations of proteoglycans.

Chondroitin sulphate-guided construction of polypyrrole nanoarchitectures

Energy Technology Data Exchange (ETDEWEB)

Zhou, Zhengnan [School of Materials Science and Engineering, South China University of Technology, Guangzhou 510641 (China); Zhu, Wenjun [Department of Prosthodontics, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou 510055 (China); Liao, Jingwen [School of Materials Science and Engineering, South China University of Technology, Guangzhou 510641 (China); Huang, Shishu [State Key Laboratory of Oral Diseases, West China College of Stomatology, Sichuan University (China); Department of Orthopaedics and Traumatology, The University of Hong Kong (China); Chen, Junqi; He, Tianrui [School of Materials Science and Engineering, South China University of Technology, Guangzhou 510641 (China); Tan, Guoxin, E-mail: tanguoxin@126.com [Faculty of Light and Chemical, Guangdong University of Technology, Guangzhou 510006 (China); Ning, Chengyun, E-mail: imcyning@scut.edu.cn [School of Materials Science and Engineering, South China University of Technology, Guangzhou 510641 (China)

2015-03-01

Nanospheres, nanocones, and nanowires are three typical polypyrrole (PPy) nanoarchitectures and electrochemically polymerized with the dope of chondroitin sulphate (CS) in this study. CS, a functional biomacromolecule, guides the formation of PPy nanoarchitectures as the dopant and morphology-directing agent. Combined with our previous reported other PPy nanoarchitectures (such as nanotube arrays and nanowires), this work further proposed the novel mechanism of the construction of PPy/CS nanoarchitectures with the synergistic effect of CS molecular chains structure and the steric hindrance. Compared to the undoped PPy, MC3T3-E1 cells with PPy/CS nanoarchitectures possessed stronger proliferation and osteogenic differentiation capability. This suggests that PPy/CS nanoarchitectures have appropriate biocompatibility. Altogether, the nanoarchitectured PPy/CS may find application in the regeneration of bone defect. - Highlights: • The formation mechanism of PPy nanoarchitectures was proposed. • CS acted as biofunctional dopant and morphology-directing agent in PPy forming. • PPy-CS nanoarchitectures were dependent on the Py/CS ratio.

Prediction of the oversulphated chondroitin sulphate contamination of unfractionated heparin by ATR-IR spectrophotometry.

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Norwig, J; Beyer, T; Brinz, D; Holzgrabe, U; Diller, M; Manns, D

2009-03-01

The detection of a contamination of heparin with oversulphated chondroitin sulphate (OSCS) was first analysed in an unfractionated heparin batch supplied to the US API-market in April 2006. OSCS is a semi-synthetic derivative of the natural occuring glycosaminoglycan chondroitin sulphate. Moreover some spectroscopic characteristics of the substance overlap with those of heparin, so that the infrared (IR) spectra are visually difficult to distinguish whereas (1)H-NMR (Nuclear Magnetic Resonance) spectroscopy or capillary electrophoresis (CE) provides identification by a simple visual inspection of either the spectrum or the electropherogram respectively. However, applying special tools of Multivariate Data Analysis (MVA) to the IR spectra an identification of the contaminated samples is possible. In detail a rapid Attenuation Total Reflectance-Infrared (ATR-IR) measurement was selected, which does not require any sample preparation. The result (contaminated or not contaminated) is predicted within a few minutes. A method transfer to mobile ATR-IR spectrometers seems to be possible. The analysis is based on the fact that the fingerprint of the OSCS IR spectrum (1st derivative) complies with a theoretically calculated principal component in the MVA.

Formulation and Evaluation of Chitosan-Chondroitin Sulphate Based Nasal Inserts for Zolmitriptan

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Kirandeep Kaur

2013-01-01

Full Text Available Bioadhesive nasal dosage forms are an attractive method for overcoming rapid mucociliary clearance transport in the nose and for delivering the drug directly to brain. The present study was designed to formulate chondroitin sulphate (CS and chitosan (CH nasal inserts employing zolmitriptan, an antimigraine drug. The interpolymer complexes (IPC formed between –COO− and – groups of CS and group of CH were characterized by infrared spectroscopy (IR, differential scanning analysis (DSC, and zeta potential studies. The unloaded and loaded nasal inserts were evaluated for water uptake studies, and bioadhesive strength studies, scanning electron microscopic studies (SEM. The in vitro drug release and in situ permeation studies were carried out on loaded nasal inserts. The DSC and IR studies confirmed the formation of a complex between the two polymers. The results indicated that the formulation F1 (CH : CS; 30 : 70 was demonstrating the highest bioadhesive strength and zeta potential. The presence of porous structure in the nasal inserts was confirmed by the SEM analysis. Further, in vitro and in situ release studies demonstrated that formulations F9 and F11 (drug : polymer; 1 : 10 were releasing 90% and 98% zolmitriptan over a period of 8 h. It can be concluded that nasal inserts formulated from chitosan-chondroitin sulphate (CH-CS interpolymer complex (IPC can be used for delivery of antimigraine drug to brain.

Distribution of radioactivity in the mouse organism after administration of 35S-chondroitin sulphate

International Nuclear Information System (INIS)

Konador, A.; Kawiak, J.

1976-01-01

The follow-up of chondroitin sulphate (ChS) distribution in the organism after its administration by various routes is interesting in view of the possibility of clinical applications. Mice received intraperitoneally and intragastrically 35 S-ChS and the distribution of radioactivity was followed in 10 chosen organs. The dynamics of changes in radioactivity were found to differ in dependence on the type of organ and route of administration. (author)

Determination of Oversulphated Chondroitin Sulphate and Dermatan Sulphate in unfractionated heparin by (1)H-NMR - Collaborative study for quantification and analytical determination of LoD.

Science.gov (United States)

McEwen, I; Mulloy, B; Hellwig, E; Kozerski, L; Beyer, T; Holzgrabe, U; Wanko, R; Spieser, J-M; Rodomonte, A

2008-12-01

Oversulphated Chondroitin Sulphate (OSCS) and Dermatan Sulphate (DS) in unfractionated heparins can be identified by nuclear magnetic resonance spectrometry (NMR). The limit of detection (LoD) of OSCS is 0.1% relative to the heparin content. This LoD is obtained at a signal-to-noise ratio (S/N) of 2000:1 of the heparin methyl signal. Quantification is best obtained by comparing peak heights of the OSCS and heparin methyl signals. Reproducibility of less than 10% relative standard deviation (RSD) has been obtained. The accuracy of quantification was good.

Proteoglycan metabolism associated with mouse metanephric development: morphologic and biochemical effects of beta-D-xyloside

International Nuclear Information System (INIS)

Platt, J.L.; Brown, D.M.; Granlund, K.; Oegema, T.R.; Klein, D.J.

1987-01-01

Morphology and de novo incorporation of [ 35 S]sulfate into proteoglycans were studied in fetal mouse kidneys at the onset of organogenesis. Branching morphogenesis and nephron development in organ culture and in vivo were associated with de novo synthesis of chondroitin-SO 4 and heparan-SO 4 proteoglycans. The role of proteoglycan metabolism in metanephrogenesis was then studied by analysis of the effects of p-nitrophenyl-beta-D-xylopyranoside (beta-D-xyloside) on renal development and proteoglycan metabolism. Incubation of fetal kidneys in beta-D-xyloside at concentrations of 1.0 and 0.5 mM, but not at 0.1 mM, caused inhibition of ureteric branching and markedly diminished synthesis of a large Mr 2.0 X 10(6) Da chondroitin-SO 4 proteoglycan. Incorporation of [ 35 S]sulfate was stimulated at all beta-D-xyloside concentrations, reflecting synthesis of xyloside initiated dermatan- 35 SO 4 chains. In contrast to dramatic effects on chondroitin-SO 4 synthesis and ureteric branching, beta-D-xyloside had no effect on heparan-SO 4 synthesis or on development of the glomerulus and glomerular basement membrane. We thus characterize the proteoglycans synthesized early in the course of renal organogenesis and describe observations which suggest an association between metabolism of chondroitin-SO 4 proteoglycan and development of the ureter

Age-related changes in the incorporation of [35S]sulfate into two proteoglycan populations from human cartilage

International Nuclear Information System (INIS)

Triphaus, G.F.; Schmidt, A.; Buddecke, E.

1980-01-01

From human hyaline cartilage (processus xyphoid) preincubated in the presence of [ 35 S] sulfate, proteoglycans were extracted by 4M guanidinium chloride and divided into 6 age groups. Fractionation of proteoglycans by gel filtration under dissociative conditions resulted in two proteoglycan fractions (a and b) with different hydrodynamic volumes. The higher molecular weight fraction a contained chondroitin sulfate, the fraction b keratan sulfate as predominant glycosaminoglycan, the chondroitin sulfate/keratan sulfate ratio decreasing with increasing age in either fraction. The relative portion of proteoglycan fraction b and its 35 S-labelling increased with increasing age. From the specific 35 S radioactivities of the chondroitin sulfate and keratan sulfate preparations, the occurrence of two independent proteoglycan populations is suggested. A precursorproduct relationship between proteoglycan fraction a and b could be excluded. (orig.)

Age-related changes in the incorporation of (/sup 35/S)sulfate into two proteoglycan populations from human cartilage

Energy Technology Data Exchange (ETDEWEB)

Triphaus, G.F.; Schmidt, A.; Buddecke, E.

1980-12-01

From human hyaline cartilage (processus xyphoid) preincubated in the presence of (/sup 35/S) sulfate, proteoglycans were extracted by 4M guanidinium chloride and divided into 6 age groups. Fractionation of proteoglycans by gel filtration under dissociative conditions resulted in two proteoglycan fractions (a and b) with different hydrodynamic volumes. The higher molecular weight fraction a contained chondroitin sulfate, the fraction b keratan sulfate as predominant glycosaminoglycan, the chondroitin sulfate/keratan sulfate ratio decreasing with increasing age in either fraction. The relative portion of proteoglycan fraction b and its /sup 35/S-labelling increased with increasing age. From the specific /sup 35/S radioactivities of the chondroitin sulfate and keratan sulfate preparations, the occurrence of two independent proteoglycan populations is suggested. A precursorproduct relationship between proteoglycan fraction a and b could be excluded.

Histamine and chondroitin sulfate E proteoglycan released by cultured human colonic mucosa: indication for possible presence of E mast cells

International Nuclear Information System (INIS)

Eliakim, R.; Gilead, L.; Ligumsky, M; Okon, E.; Rachmilewitz, D.; Razin, E.

1986-01-01

An association between the release of histamine and chondroitin sulfate E proteoglycan (PG) was demonstrates in human colonic mucosa (HCM). Colonic biopsy samples incorporated [ 35 S]sulfate into PG, which was partially released into the culture medium during the incubation period. Ascending thin-layer chromatography of the released 35 S-labeled PG after its digestion by chondroitin ABC lyase (chondroitinase, EC 4.2.2.4) followed by autoradiography yielded three products that migrated in the position of monosulfated disaccharides of N-acetylgalactosamine 4-sulfate and N-acetylgalactosoamine 6-sulfate and of an oversulfated disaccharide possessing N-acetylgalatosamine 4,6-disulfate. Cultured colonic mucosa released 23.6 +/- 3.7ng of histamine per mg of wet tissue without any special trigger. Comparison by linear regression analysis of the release of histamine and chondroitin [ 35 S]sulfate E PG revealed a correlation coefficient (r) of 0.7. Histological examination of the colonic biopsies revealed the presence of many mast cells in various degrees of degranulation in the mucosa and submucosa. The above correlation, the observation that most of the mast cells showed various degrees of degranulation, and the lack of heparin synthesis as opposed to the synthesis and immunological release of chondroitin sulfate E strongly suggest that the E mast cell exists in the human colon

Proteoglycan biosynthesis in murine monocytic leukemic (M1) cells before and after differentiation

International Nuclear Information System (INIS)

McQuillan, D.J.; Yanagishita, M.; Hascall, V.C.; Bickel, M.

1989-01-01

Murine monocytic leukemic (M1) cells were cultured in the presence of [ 3 H]glucosamine and [ 35 S]sulfate. Labeled proteoglycans were purified by anion exchange chromatography and characterized by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis in combination with chemical and enzymatic degradation. M1 cells synthesize a single predominant species of proteoglycan which distributes almost equally between the cell and medium after 17 h labeling. The cell-associated proteoglycan has an overall size of about 135 kDa and contains three to five chondroitin sulfate chains (28-31 kDa each) attached to a chondroitinase-generated core protein of 28 kDa. The synthesis and subsequent secretion of this proteoglycan was enhanced 4-5-fold in cells induced to differentiate into macrophages. This was not a phenomenon of arrest in the G0/G1 stage of the cell cycle, since density inhibited undifferentiated cells arrested at this stage did not increase proteoglycan synthesis. The chondroitin sulfate chains contained exclusively chondroitin 4- and 6-sulfate; however, the ratio of these two disaccharides differed between the medium- and cell-associated proteoglycans, and changed during progression of the cells into a fully differentiated phenotype. Pulse-chase kinetics indicate the presence of two distinct pools of proteoglycan; one that is secreted very rapidly from the cell after a approximately 1-h lag, and a second pool that is turned over in the cell with a half-time of approximately 3.5 h. Subtle differences in the glycosylation patterns of the medium- and cell-associated species are consistent with synthesis of two pools. Papain digestion suggests that the chondroitin sulfate chains are clustered on a small protease resistant peptide. The data suggest that this proteoglycan is similar to the serglycin proteoglycan family

Structural studies on sulfated oligosaccharides derived from the carbohydrate-protein linkage region of chondroitin 6-sulfate proteoglycans of shark cartilage. (II.) Seven compounds containing 2 or 3 sulfate residues.

NARCIS (Netherlands)

Vliegenthart, J.F.G.; Waard, P. de; Harada, T.; Sugahara, K.

1992-01-01

Shark cartilage proteoglycans bear predominantly chondroitin 6-sulfate. After exhaustive protease digestion, reductive beta-elimination and subsequent chondroitinase ABC digestion, 13 hexasaccharide alditols were obtained from the carbohydrate-protein linkage region and six of them contain 0 or 1

Intravesical Glycosaminoglycan Replacement with Chondroitin Sulphate (Gepan? instill) in Patients with Chronic Radiotherapy- or Chemotherapy-Associated Cystitis

OpenAIRE

Schwalenberg, Thilo; Berger, Frank Peter; Horn, Lars Christian; Thi, Phuc Ho; Stolzenburg, Jens?Uwe; Neuhaus, Jochen

2015-01-01

Background and Objective Intravesical instillation of glycosaminoglycans is a promising option for the treatment of chronic cystitis, as it supports the regeneration of the damaged urothelial layer. We investigated the efficacy of short-term intravesical chondroitin sulphate treatment (six courses of instillation) in patients with chronic radiotherapy- or chemotherapy-associated cystitis. Methods This prospective, observational study included patients with chronic radiotherapy- or chemotherap...

A novel multistep method for chondroitin sulphate immobilization and its interaction with fibroblast cells

Energy Technology Data Exchange (ETDEWEB)

Ozaltin, Kadir; Lehocký, Marián, E-mail: lehocky@post.cz; Kuceková, Zdenka; Humpolíček, Petr; Sáha, Petr

2017-01-01

Polymeric biomaterials are widely used in medical applications owing to their low cost, processability and sufficient toughness. Surface modification by creating a thin film of bioactive agents is promising technique to enhance cellular interactions, regulate the protein adsorption and/or avoid bacterial infections. Polyethylene is one of the most used polymeric biomaterial but its hydrophobic nature impedes its further chemical modifications. Plasma treatment is unique method to increase its hydrophilicity by incorporating hydrophilic oxidative functional groups and tailoring the surface by physical etching. Furthermore, grafting of polymer brushes of amine group containing monomers onto the functionalized surface lead to strongly immobilized bioactive agents at the final step. Chondroitin sulphate is natural polysaccharide mainly found in connective cartilage tissue which used as a bioactive agent to immobilize onto polyethylene surface by multistep method in this study. - Highlights: • Attachment of chondroitin sulfate to polyethylene. • A robust way to modify surfaces using multistep approach. • The modified surfaces showed improved proliferation of mouse primary fibroblast cells.

Chondroitin Sulfate Perlecan Enhances Collagen Fibril Formation

DEFF Research Database (Denmark)

Kvist, A. J.; Johnson, A. E.; Mörgelin, M.

2006-01-01

in collagen type II fibril assembly by perlecan-null chondrocytes. Cartilage perlecan is a heparin sulfate or a mixed heparan sulfate/chondroitin sulfate proteoglycan. The latter form binds collagen and accelerates fibril formation in vitro, with more defined fibril morphology and increased fibril diameters...... produced in the presence of perlecan. Interestingly, the enhancement of collagen fibril formation is independent on the core protein and is mimicked by chondroitin sulfate E but neither by chondroitin sulfate D nor dextran sulfate. Furthermore, perlecan chondroitin sulfate contains the 4,6-disulfated...... disaccharides typical for chondroitin sulfate E. Indeed, purified glycosaminoglycans from perlecan-enriched fractions of cartilage extracts contain elevated levels of 4,6-disulfated chondroitin sulfate disaccharides and enhance collagen fibril formation. The effect on collagen assembly is proportional...

Digestion of proteoglycan by Bacteroides thetaiotaomicron.

OpenAIRE

Kuritza, A P; Salyers, A A

1983-01-01

It has been shown previously that Bacteroides thetaiotaomicron, a human colonic anaerobe, can utilize the tissue mucopolysaccharide chondroitin sulfate as a source of carbon and energy and that the enzymes involved in this utilization are all cell associated (A. A. Salyers and M. B. O'Brien, J. Bacteriol. 143:772-780, 1980). Since chondroitin sulfate does not generally occur in isolated form in tissue, but rather is bound covalently in proteoglycan, we investigated the extent to which chondro...

Production and characterization of monoclonal antibodies directed against connective tissue proteoglycans

DEFF Research Database (Denmark)

Caterson, B; Christner, J E; Baker, J R

1985-01-01

distribution of 4- and 6-sulfated and unsulfated proteoglycans in tissue sections of cartilage and other noncartilaginous tissues. Digestion with chondroitinase ABC or ACII can be used to differentiate between chondroitin sulfate and dermatan sulfate proteoglycan in different connective tissues. In addition...

Reduced sulfation of chondroitin sulfate but not heparan sulfate in kidneys of diabetic db/db mice.

Science.gov (United States)

Reine, Trine M; Grøndahl, Frøy; Jenssen, Trond G; Hadler-Olsen, Elin; Prydz, Kristian; Kolset, Svein O

2013-08-01

Heparan sulfate proteoglycans are hypothesized to contribute to the filtration barrier in kidney glomeruli and the glycocalyx of endothelial cells. To investigate potential changes in proteoglycans in diabetic kidney, we isolated glycosaminoglycans from kidney cortex from healthy db/+ and diabetic db/db mice. Disaccharide analysis of chondroitin sulfate revealed a significant decrease in the 4-O-sulfated disaccharides (D0a4) from 65% to 40%, whereas 6-O-sulfated disaccharides (D0a6) were reduced from 11% to 6%, with a corresponding increase in unsulfated disaccharides. In contrast, no structural differences were observed in heparan sulfate. Furthermore, no difference was found in the molar amount of glycosaminoglycans, or in the ratio of hyaluronan/heparan sulfate/chondroitin sulfate. Immunohistochemical staining for the heparan sulfate proteoglycan perlecan was similar in both types of material but reduced staining of 4-O-sulfated chondroitin and dermatan was observed in kidney sections from diabetic mice. In support of this, using qRT-PCR, a 53.5% decrease in the expression level of Chst-11 (chondroitin 4-O sulfotransferase) was demonstrated in diabetic kidney. These results suggest that changes in the sulfation of chondroitin need to be addressed in future studies on proteoglycans and kidney function in diabetes.

The endogenous proteoglycan-degrading enzyme ADAMTS-4 promotes functional recovery after spinal cord injury

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Tauchi Ryoji

2012-03-01

Full Text Available Abstract Background Chondroitin sulfate proteoglycans are major inhibitory molecules for neural plasticity under both physiological and pathological conditions. The chondroitin sulfate degrading enzyme chondroitinase ABC promotes functional recovery after spinal cord injury, and restores experience-dependent plasticity, such as ocular dominance plasticity and fear erasure plasticity, in adult rodents. These data suggest that the sugar chain in a proteoglycan moiety is essential for the inhibitory activity of proteoglycans. However, the significance of the core protein has not been studied extensively. Furthermore, considering that chondroitinase ABC is derived from bacteria, a mammalian endogenous enzyme which can inactivate the proteoglycans' activity is desirable for clinical use. Methods The degradation activity of ADAMTS-4 was estimated for the core proteins of chondroitin sulfate proteoglycans, that is, brevican, neurocan and phosphacan. To evaluate the biological significance of ADMATS-4 activity, an in vitro neurite growth assay and an in vivo neuronal injury model, spinal cord contusion injury, were employed. Results ADAMTS-4 digested proteoglycans, and reversed their inhibition of neurite outgrowth. Local administration of ADAMTS-4 significantly promoted motor function recovery after spinal cord injury. Supporting these findings, the ADAMTS-4-treated spinal cord exhibited enhanced axonal regeneration/sprouting after spinal cord injury. Conclusions Our data suggest that the core protein in a proteoglycan moiety is also important for the inhibition of neural plasticity, and provides a potentially safer tool for the treatment of neuronal injuries.

Human glomerular epithelial cell proteoglycans

International Nuclear Information System (INIS)

Thomas, G.J.; Jenner, L.; Mason, R.M.; Davies, M.

1990-01-01

Proteoglycans synthesized by cultures of human glomerular epithelial cells have been isolated and characterized. Three types of heparan sulfate were detected. Heparan sulfate proteoglycan I (HSPG-I; Kav 6B 0.04) was found in the cell layer and medium and accounted for 12% of the total proteoglycans synthesized. HSPG-II (Kav 6B 0.25) accounted for 18% of the proteoglycans and was located in the medium and cell layer. A third population (9% of the proteoglycan population), heparan sulfate glycosaminoglycan (HS-GAG; Kav 6B 0.4-0.8), had properties consistent with single glycosaminoglycan chains or their fragments and was found only in the cell layer. HSPG-I and HSPG-II from the cell layer had hydrophobic properties; they were released from the cell layer by mild trypsin treatment. HS-GAG lacked these properties, consisted of low-molecular-mass heparan sulfate oligosaccharides, and were intracellular. HSPG-I and -II released to the medium lacked hydrophobic properties. The cells also produced three distinct types of chondroitin sulfates. The major species, chondroitin sulfate proteoglycan I (CSPG-I) eluted in the excluded volume of a Sepharose CL-6B column, accounted for 30% of the proteoglycans detected, and was found in both the cell layer and medium. Cell layer CSPG-I bound to octyl-Sepharose. It was released from the cell layer by mild trypsin treatment. CSPG-II (Kav 6B 0.1-0.23) accounted for 10% of the total 35S-labeled macromolecules and was found predominantly in the culture medium. A small amount of CS-GAG (Kav 6B 0.25-0.6) is present in the cell extract and like HS-GAG is intracellular. Pulse-chase experiments indicated that HSPG-I and -II and CSPG-I and -II are lost from the cell layer either by direct release into the medium or by internalization where they are metabolized to single glycosaminoglycan chains and subsequently to inorganic sulfate

Chondroitin Sulfate Proteoglycan 4 and Its Potential As an Antibody Immunotherapy Target across Different Tumor Types

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Kristina M. Ilieva

2018-01-01

Full Text Available Overexpression of the chondroitin sulfate proteoglycan 4 (CSPG4 has been associated with the pathology of multiple types of such as melanoma, breast cancer, squamous cell carcinoma, mesothelioma, neuroblastoma, adult and pediatric sarcomas, and some hematological cancers. CSPG4 has been reported to exhibit a role in the growth and survival as well as in the spreading and metastasis of tumor cells. CSPG4 is overexpressed in several malignant diseases, while it is thought to have restricted and low expression in normal tissues. Thus, CSPG4 has become the target of numerous anticancer treatment approaches, including monoclonal antibody-based therapies. This study reviews key potential anti-CSPG4 antibody and immune-based therapies and examines their direct antiproliferative/metastatic and immune activating mechanisms of action.

Molecular polymorphism of a cell surface proteoglycan: distinct structures on simple and stratified epithelia.

Science.gov (United States)

Sanderson, R D; Bernfield, M

1988-12-01

Epithelial cells are organized into either a single layer (simple epithelia) or multiple layers (stratified epithelia). Maintenance of these cellular organizations requires distinct adhesive mechanisms involving many cell surface molecules. One such molecule is a cell surface proteoglycan, named syndecan, that contains both heparan sulfate and chondroitin sulfate chains. This proteoglycan binds cells to fibrillar collagens and fibronectin and thus acts as a receptor for interstitial matrix. The proteoglycan is restricted to the basolateral surface of simple epithelial cells, but is located over the entire surface of stratified epithelial cells, even those surfaces not contacting matrix. We now show that the distinct localization in simple and stratified epithelia correlates with a distinct proteoglycan structure. The proteoglycan from simple epithelia (modal molecular size, 160 kDa) is larger than that from stratified epithelia (modal molecular size, 92 kDa), but their core proteins are identical in size and immunoreactivity. The proteoglycan from simple epithelia has more and larger heparan sulfate and chondroitin sulfate chains than the proteoglycan from stratified epithelia. Thus, the cell surface proteoglycan shows a tissue-specific structural polymorphism due to distinct posttranslational modifications. This polymorphism likely reflects distinct proteoglycan functions in simple and stratified epithelia, potentially meeting the different adhesive requirements of the cells in these different organizations.

Perlecan and basement membrane-chondroitin sulfate proteoglycan (bamacan) are two basement membrane chondroitin/dermatan sulfate proteoglycans in the Engelbreth-Holm-Swarm tumor matrix

DEFF Research Database (Denmark)

Couchman, J R; Kapoor, R; Sthanam, M

1996-01-01

heparan sulfate proteoglycan, widespread in many basement membranes and connective tissues. We now identify two distinct proteoglycan species from this tumor source, which are substituted with galactosaminoglycans and which show basement membrane localization by immunohistochemistry. One species......The presence of proteoglycans bearing galactosaminoglycan chains has been reported, but none has been identified previously in the matrix of the Engelbreth-Holm-Swarm tumor, which is a source of several basement membrane components. This tumor matrix contains perlecan, a large, low buoyant density......-CSPG are distinct in core protein structure. Both are, however, basement membrane components, although there are tissue-specific differences in their distribution....

Distribution of two basement membrane proteoglycans through hair follicle development and the hair growth cycle in the rat

DEFF Research Database (Denmark)

Couchman, J R; King, J L; McCarthy, K J

1990-01-01

The distribution of two distinct populations of basement membrane proteoglycans has been monitored through hair growth development in the rat embryo and subsequent hair growth cycle. An antiserum against a small heparan sulfate proteoglycan uniformly stained the dermal-epidermal junction...... of embryonic rats throughout the period of hair follicle formation. On the other hand, monoclonal antibodies recognizing a basement membrane-specific chondroitin sulfate proteoglycan only weakly stained 16-d embryo dermal-epidermal junction, but strong staining was associated with hair follicle buds...... as they developed. Through the hair growth cycle, it was found that the heparan sulfate proteoglycan persisted around the follicles, while the chondroitin sulfate proteoglycan decreased in amount through catagen until it was undetectable at the base and dermal papilla of the telogen follicle. As anagen commenced...

Differential expression of proteoglycans in tissue remodeling and lymphangiogenesis after experimental renal transplantation in rats.

Directory of Open Access Journals (Sweden)

Heleen Rienstra

Full Text Available BACKGROUND: Chronic transplant dysfunction explains the majority of late renal allograft loss and is accompanied by extensive tissue remodeling leading to transplant vasculopathy, glomerulosclerosis and interstitial fibrosis. Matrix proteoglycans mediate cell-cell and cell-matrix interactions and play key roles in tissue remodeling. The aim of this study was to characterize differential heparan sulfate proteoglycan and chondroitin sulfate proteoglycan expression in transplant vasculopathy, glomerulosclerosis and interstitial fibrosis in renal allografts with chronic transplant dysfunction. METHODS: Renal allografts were transplanted in the Dark Agouti-to-Wistar Furth rat strain combination. Dark Agouti-to-Dark Agouti isografts and non-transplanted Dark Agouti kidneys served as controls. Allograft and isograft recipients were sacrificed 66 and 81 days (mean after transplantation, respectively. Heparan sulfate proteoglycan (collXVIII, perlecan and agrin and chondroitin sulfate proteoglycan (versican expression, as well as CD31 and LYVE-1 (vascular and lymphatic endothelium, respectively expression were (semi- quantitatively analyzed using immunofluorescence. FINDINGS: Arteries with transplant vasculopathy and sclerotic glomeruli in allografts displayed pronounced neo-expression of collXVIII and perlecan. In contrast, in interstitial fibrosis expression of the chondroitin sulfate proteoglycan versican dominated. In the cortical tubular basement membranes in both iso- and allografts, induction of collXVIII was detected. Allografts presented extensive lymphangiogenesis (p<0.01 compared to isografts and non-transplanted controls, which was associated with induced perlecan expression underneath the lymphatic endothelium (p<0.05 and p<0.01 compared to isografts and non-transplanted controls, respectively. Both the magnitude of lymphangiogenesis and perlecan expression correlated with severity of interstitial fibrosis and impaired graft function

Three distinct molecular species of proteoglycan synthesized by the rat limb bud at the prechondrogenic stage

International Nuclear Information System (INIS)

Matsui, F.; Oohira, A.; Shoji, R.; Nogami, H.

1989-01-01

To characterize proteoglycans in the prechondrogenic limb bud, proteoglycans were extracted with 4 M guanidine HCl containing a detergent and protease inhibitors from Day 13 fetal rat limb buds which had been labeled with [35S]sulfate for 3 h in vitro. About 90% of 35S-labeled proteoglycans was solubilized under the conditions used. The proteoglycan preparation was separated by DEAE-Sephacel column chromatography into three peaks; peak I eluted at 0.45 M NaCl concentration, peak II at 0.52 M, and peak III at 1.4 M. Peaks I and III were identified as proteoglycans bearing heparan sulfate side chains. The heparan sulfate proteoglycan in peak III was larger in hydrodynamic size than the proteoglycan in peak I. The heparan sulfate side chains of peak III proteoglycan were smaller in the size and more abundant in N-sulfated glucosamine than those of peak I proteoglycan. Peak II contained a chondroitin sulfate proteoglycan with a core protein of a doublet of Mr 550,000 and 500,000. The chondroitin sulfate proteoglycan was easily solubilized with a physiological salt solution and the heparan sulfate proteoglycan in peak I was partially solubilized with the physiological salt solution. The remainder of the proteoglycan in peak I and the heparan sulfate proteoglycan in peak III could be solubilized effectively only with a solution containing a detergent, such as nonanoyl-N-methylglucamide. This observation indicates the difference in the localization among these three proteoglycans in the developing rat limb bud

Iduronic Acid in chondroitin/dermatan sulfate affects directional migration of aortic smooth muscle cells

NARCIS (Netherlands)

Bartolini, B.; Thelin, M.A.; Svensson, L.; Ghiselli, G.; Kuppevelt, T.H. van; Malmstrom, A.; Maccarana, M.

2013-01-01

Aortic smooth muscle cells produce chondroitin/dermatan sulfate (CS/DS) proteoglycans that regulate extracellular matrix organization and cell behavior in normal and pathological conditions. A unique feature of CS/DS proteoglycans is the presence of iduronic acid (IdoA), catalyzed by two DS

Biphasic Role of Chondroitin Sulfate in Cardiac Differentiation of Embryonic Stem Cells through Inhibition of Wnt/beta-Catenin Signaling

NARCIS (Netherlands)

Prinz, R.D.; Willis, C.M.; Kuppevelt, T.H. van; Kluppel, M.

2014-01-01

The glycosaminoglycan chondroitin sulfate is a critical component of proteoglycans on the cell surface and in the extracellular matrix. As such, chondroitin sulfate side chains and the sulfation balance of chondroitin play important roles in the control of signaling pathways, and have a functional

Purification and partial characterization of glycosaminoglycans and proteoglycans from cultured rabbit smooth muscle cells

International Nuclear Information System (INIS)

Sabatino, R.D.

1985-01-01

Glycosaminoglycans synthesized by cultured rabbit smooth muscle cells were isolated after incorporation of [ 3 H]-glucosamine into glycosaminoglycans in the presence or absence of 10% fetal bovine serum. Glycosaminoglycans were quantitated by two-dimensional electrophoresis after proteolytic digestion of the cell layers and media. The results show that the presence of serum has no effect on the chondroitin sulfate, heparan sulfate and dermatan sulfate content of the cell layers. The incorporation of [ 3 H]-glucosamine into hyaluronic acid of the cell layers was three times higher in the presence of serum. In the medium , the quantity of hyaluronic was two times higher in the presence of serum while the other glycosaminoglycans remained unchanged. The incorporation of [ 3 H]-glucosamine into hyaluronic acid was unaffected by the presence of serum. Specific proteoglycans were isolated from medium after with [ 35 S]-sulfate and [ 3 H]-serine by isopycnic ultracentrifugation and chromatography on Sepharose CL-4B and DEAE-cellulose. Preparations contained a chondroitin sulfate proteoglycan, a condroitin sulfate-dermatan sulfate proteoglycan and a heparan sulfate proteoglycan. Glycosaminoglycans and proteoglycans synthesized by rabbit aorta smooth muscle cells are similar to those from human aorta

Macrophage secretory products selectively stimulate dermatan sulfate proteoglycan production in cultured arterial smooth muscle cells

International Nuclear Information System (INIS)

Edwards, I.J.; Wagner, W.D.; Owens, R.T.

1990-01-01

Arterial dermatan sulfate proteoglycan has been shown to increase with atherosclerosis progression, but factors responsible for this increase are unknown. To test the hypothesis that smooth muscle cell proteoglycan synthesis may be modified by macrophage products, pigeon arterial smooth muscle cells were exposed to the media of either cholesteryl ester-loaded pigeon peritoneal macrophages or a macrophage cell line P388D1. Proteoglycans radiolabeled with [35S]sulfate and [3H]serine were isolated from culture media and smooth muscle cells and purified following precipitation with 1-hexadecylpyridinium chloride and chromatography. Increasing concentrations of macrophage-conditioned media were associated with a dose-response increase in [35S]sulfate incorporation into secreted proteoglycans, but there was no change in cell-associated proteoglycans. Incorporation of [3H]serine into total proteoglycan core proteins was not significantly different (5.2 X 10(5) dpm and 5.5 X 10(5) disintegrations per minute (dpm) in control and conditioned media-treated cultures, respectively), but selective effects were observed on individual proteoglycan types. Twofold increases in dermatan sulfate proteoglycan and limited degradation of chondroitin sulfate proteoglycan were apparent based on core proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Immunoinhibition studies indicated that interleukin-1 was involved in the modulation of proteoglycan synthesis by macrophage-conditioned media. These data provide support for the role of macrophages in alteration of the matrix proteoglycans synthesized by smooth muscle cells and provide a mechanism to account for the reported increased dermatan sulfate/chondroitin sulfate ratios in the developing atherosclerotic lesion

Axonal transport of proteoglycans to the goldfish optic tectum

International Nuclear Information System (INIS)

Ripellino, J.A.; Elam, J.S.

1988-01-01

The study addressed the question of whether 35 SO 4 labeled molecules that have been delivered to the goldfish optic nerve terminals by rapid axonal transport include soluble proteoglycans. For analysis, tectal homogenates were subfractionated into a soluble fraction (soluble after centrifugation at 105,000 g), a lysis fraction (soluble after treatment with hypotonic buffer followed by centrifugation at 105,000 g) and a final 105,000 g pellet fraction. The soluble fraction contained 25.7% of incorporated radioactivity and upon DEAE chromatography was resolved into a fraction of sulfated glycoproteins eluting at 0-0.32 M NaCl and containing 39.5% of total soluble label and a fraction eluting at 0.32-0.60 M NaCl containing 53.9% of soluble label. This latter fraction was included on columns of Sepharose CL-6B with or without 4 M guanidine and after pronase digestion was found to have 51% of its radioactivity contained in the glycosaminoglycans (GAGs) heparan sulfate and chondroitin (4 or 6) sulfate in the ratio of 70% to 30%. Mobility of both intact proteoglycans and constituent GAGs on Sepharose CL-6B indicated a size distribution that is smaller than has been observed for proteoglycans and GAGs from cultured neuronal cell lines. Similar analysis of lysis fraction, containing 11.5% of incorporated 35 SO 4 , showed a mixture of heparan sulfate and chondroitin sulfate containing proteoglycans, apparent free heparan sulfate and few, if any, sulfated glycoproteins. Overall, the results support the hypothesis that soluble proteoglycans are among the molecules axonally transported in the visual system

MDA-MB-231 breast cancer cell viability, motility and matrix adhesion are regulated by a complex interplay of heparan sulfate, chondroitin-/dermatan sulfate and hyaluronan biosynthesis.

Science.gov (United States)

Viola, Manuela; Brüggemann, Kathrin; Karousou, Evgenia; Caon, Ilaria; Caravà, Elena; Vigetti, Davide; Greve, Burkhard; Stock, Christian; De Luca, Giancarlo; Passi, Alberto; Götte, Martin

2017-06-01

Proteoglycans and glycosaminoglycans modulate numerous cellular processes relevant to tumour progression, including cell proliferation, cell-matrix interactions, cell motility and invasive growth. Among the glycosaminoglycans with a well-documented role in tumour progression are heparan sulphate, chondroitin/dermatan sulphate and hyaluronic acid/hyaluronan. While the mode of biosynthesis differs for sulphated glycosaminoglycans, which are synthesised in the ER and Golgi compartments, and hyaluronan, which is synthesized at the plasma membrane, these polysaccharides partially compete for common substrates. In this study, we employed a siRNA knockdown approach for heparan sulphate (EXT1) and heparan/chondroitin/dermatan sulphate-biosynthetic enzymes (β4GalT7) in the aggressive human breast cancer cell line MDA-MB-231 to study the impact on cell behaviour and hyaluronan biosynthesis. Knockdown of β4GalT7 expression resulted in a decrease in cell viability, motility and adhesion to fibronectin, while these parameters were unchanged in EXT1-silenced cells. Importantly, these changes were associated with a decreased expression of syndecan-1, decreased signalling response to HGF and an increase in the synthesis of hyaluronan, due to an upregulation of the hyaluronan synthases HAS2 and HAS3. Interestingly, EXT1-depleted cells showed a downregulation of the UDP-sugar transporter SLC35D1, whereas SLC35D2 was downregulated in β4GalT7-depleted cells, indicating an intricate regulatory network that connects all glycosaminoglycans synthesis. The results of our in vitro study suggest that a modulation of breast cancer cell behaviour via interference with heparan sulphate biosynthesis may result in a compensatory upregulation of hyaluronan biosynthesis. These findings have important implications for the development of glycosaminoglycan-targeted therapeutic approaches for malignant diseases.

Proteoglycon synthesis by articular chondrocytes in agarose culture

International Nuclear Information System (INIS)

Sweet, M.B.E.; Grisillo, A.; Coehlo, A.; Schnitzler, C.M.

1987-01-01

Articular chondrocytes were isolated from knee joints of full-term bovine foetuses and grown in long-term agarose cultures. At intervals, cultures were labelled with 35 S-[sulphate] or D[6- 3 H] glucosamine. Newly synthesized proteoglycans were extracted with 4 M guanidine HCl and purified by isopycnic density gradient centrifugation or on DEAE cellulose in the presence of 8 M urea. Characterization of the proteoglycans revealed them to be identical in size to those present in the tissue and to be similarly capable of aggregation with hyaluronate. Newly synthesized chondroitin sulphate chains were identical in size, but newly synthesized keratan sulphate chains were somewhat larger than those present in the tissue. The newly synthesized proteoglycans were shown to contain the same range of O-linked oligosaccharides identified in proteoglycans of the Swarm rat chondrosarcoma. Cartilage-specific proteoglycan continued to be synthesized by the chondrocytes for up to 60 days; however, with time, proportionately more of a small non-aggregating proteoglycan appeared

Transforming growth factor-beta 1 stimulates synthesis of proteoglycan aggregates in calf articular cartilage organ cultures

International Nuclear Information System (INIS)

Morales, T.I.

1991-01-01

Previous work showed that transforming growth factor-beta 1 (TGF-beta 1), added alone to bovine cartilage organ cultures, stimulated [35S]sulfate incorporation into macromolecular material but did not investigate the fidelity of the stimulated system to maintain synthesis of cartilage-type proteoglycans. This paper provides evidence that chondrocytes synthesize the appropriate proteoglycan matrix under TGF-beta 1 stimulation: (1) there is a coordinated increase in hyaluronic acid and proteoglycan monomer synthesis, (2) link-stable proteoglycan aggregates are assembled, (3) the hybrid chondroitin sulfate/keratan sulfate monomeric species is synthesized, and (4) there is an increase in protein core synthesis. Some variation in glycosylation patterns was observed when proteoglycans synthesized under TGF-beta 1 stimulation were compared to those synthesized under basal conditions. Thus comparing TGF-beta 1 to basal samples respectively, the monomers were larger (Kav on Sepharose CL-2B = 0.29 vs 0.41), the chondroitin sulfate chains were longer by approximately 3.5 kDa, the percentage of total glycosaminoglycan in keratan sulfate increased slightly from approximately 4% (basal) to approximately 6%, and the unsulfated disaccharide decreased from 28% (basal) to 12%. All of these variations are in the direction of a more anionic proteoglycan. Since the ability of proteoglycans to confer resiliency to the cartilage matrix is directly related to their anionic nature, these changes would presumably have a beneficial effect on tissue function

Chondroitin sulfates do not impede axonal regeneration in goldfish spinal cord.

Science.gov (United States)

Takeda, Akihito; Okada, Soichiro; Funakoshi, Kengo

2017-10-15

Chondroitin sulfate proteoglycans produced in glial scar tissue are a major inhibitory factor for axonal regeneration after central nervous system injury in mammals. The inhibition is largely due to chondroitin sulfates, whose effects differ according to the sulfation pattern. In contrast to mammals, fish nerves spontaneously regenerate beyond the scar tissue after spinal cord injury, although the mechanisms that allow for axons to pass through the scar are unclear. Here, we used immunohistochemistry to examine the expression of two chondroitin sulfates with different sulfation variants at the lesion site in goldfish spinal cord. The intact spinal cord was immunoreactive for both chondroitin sulfate-A (CS-A) and chondroitin sulfate-C (CS-C), and CS-A immunoreactivity overlapped extensively with glial processes positive for glial fibrillary acidic protein. At 1week after inducing the spinal lesion, CS-A immunoreactivity was observed in the cell bodies and extracellular matrix, as well as in glial processes surrounding the lesion center. At 2weeks after the spinal lesion, regenerating axons entering the lesion center overtook the CS-A abundant area. In contrast, at 1week after lesion induction, CS-C immunoreactivity was significantly decreased, and at 2weeks after lesion induction, CS-C immunoreactivity was observed along the regenerating axons entering the lesion center. The present findings suggest that after spinal cord injury in goldfish, chondroitin sulfate proteoglycans are deposited in the extracellular matrix at the lesion site but do not form an impenetrable barrier to the growth of regenerating axons. Copyright © 2017 Elsevier B.V. All rights reserved.

Cell-surface proteoglycan in sea urchin primary mesenchyme cell migration

International Nuclear Information System (INIS)

Lane, M.C.

1989-01-01

Early in the development of the sea urchin embryo, the primary mesenchyme cells (PMC) migrate along the basal lamina of the blastocoel. Migration is inhibited in L. pictus embryos cultured in sulfate-free seawater and in S. purpuratus embryos exposed to exogenous β-D-xylosides. An in vitro assay was developed to test the migratory capacity of normal PMC on normal and treated blastocoelic matrix. Sulfate deprivation and exposure to exogenous xyloside render PMC nonmotile on either matrix. Materials removed from the surface of normal PMC by treatment with 1 M urea restored migratory ability to defective cells, whereas a similar preparation isolated from the surface of epithelial cells at the same stage did not. Migration also resumed when cells were removed from the xyloside or returned to normal seawater. The urea extract was partially purified and characterized by radiolabeling, gel electrophoresis, fluorography, ion exchange chromatography, and western blotting. The PMC synthesize a large chondroitin sulfate/dermatan sulfate proteoglycan that is present in an active fraction isolated by chromatography. Chondroitinase ABC digestion of live cells blocked migration reversibly, further supporting the identification of the chondroitin sulfate/dermatan sulfate proteoglycan as the active component in the urea extract. Much of the incorporated sulfate was distributed along the filopodia in 35 SO 4 -labelled PMC by autoradiography. The morphology of normal and treated S. purpuratus PMC was examined by scanning electron microscopy, and differences in spreading, particularly of the extensive filopodia present on the cells, was observed. A model for the role of the chondroitin sulfate/dermatan sulfate proteoglycan in cell detachment during migration is proposed

Amino acid sequence surrounding the chondroitin sulfate attachment site of thrombomodulin regulates chondroitin polymerization.

Science.gov (United States)

Izumikawa, Tomomi; Kitagawa, Hiroshi

2015-05-01

Thrombomodulin (TM) is a cell-surface glycoprotein and a critical mediator of endothelial anticoagulant function. TM exists as both a chondroitin sulfate (CS) proteoglycan (PG) form and a non-PG form lacking a CS chain (α-TM); therefore, TM can be described as a part-time PG. Previously, we reported that α-TM bears an immature, truncated linkage tetrasaccharide structure (GlcAβ1-3Galβ1-3Galβ1-4Xyl). However, the biosynthetic mechanism to generate part-time PGs remains unclear. In this study, we used several mutants to demonstrate that the amino acid sequence surrounding the CS attachment site influences the efficiency of chondroitin polymerization. In particular, the presence of acidic residues surrounding the CS attachment site was indispensable for the elongation of CS. In addition, mutants defective in CS elongation did not exhibit anti-coagulant activity, as in the case with α-TM. Together, these data support a model for CS chain assembly in which specific core protein determinants are recognized by a key biosynthetic enzyme involved in chondroitin polymerization. Copyright © 2015 Elsevier Inc. All rights reserved.

ELISA for the core protein of the cartilage large aggregating proteoglycan, aggrecan: comparison with the concentrations of immunogenic keratan sulphate in synovial fluid, serum and urine

DEFF Research Database (Denmark)

Møller, H J; Larsen, F S; Ingemann-Hansen, T

1994-01-01

ELISA. The within-assay and between-assay coefficients of variation were 4.9-8.9% and 11.1-13.0%, respectively. The mean concentrations of core protein in synovial fluid, serum and urine were 76.4 micrograms/ml, 104.0 ng/ml and 81.0 ng/ml, respectively. In synovial fluids the concentrations were closely......Immunological assays for fragments of the cartilage large aggregating proteoglycan, aggrecan, have been widely used to monitor cartilage turnover. These assays have commonly employed the monoclonal keratan sulphate antibody, 5D4. Keratan sulphate, however, is present in many tissues and 5D4...

[Efficacy of chondroitin sulphate in the treatment of elderly patients with gonarthrosis and coxarthrosis].

Science.gov (United States)

Lazebnik, L B; Drozdov, V N

2005-01-01

To study chondroitin sulphate (CS) efficacy, tolerance and response duration in elderly patients with osteoarthrosis (OA) with consideration of OA duration and stage. A total of 97 patients aged 65-85 years with stage II-IV OA by Kellgren-Lawrence received CS treatment. The treatment efficacy was assessed by functional Lecken's index, pain intensity at walking and rest, general functional condition, need in nonsteroid anti-inflammatory (NSAI) drugs. Standardization was achieved with visual analogue scale (VAS). Positive effects (pain relief, better functional parameters, lower intake of NSAI drugs) were more pronounced and stable in patients with OA stage 1 and 2. In OA of stage 3 and 4, a beneficial effect of a 6-month CS course was unstable. Side effects were at the level of mean statistics. CS (structum) is recommended for treatment of OA stage 1-4 by Kellgren-Lawrence. Duration of the treatment depends on severity of x-ray symptoms of the disease.

Brain heparan sulphate proteoglycans are altered in developing foetus when exposed to in-utero hyperglycaemia.

Science.gov (United States)

Sandeep, M S; Nandini, C D

2017-08-01

In-utero exposure of foetus to hyperglycaemic condition affects the growth and development of the organism. The brain is one of the first organs that start to develop during embryonic period and glycosaminoglycans (GAGs) and proteoglycans (PGs) are one of the key molecules involved in its development. But studies on the effect of hyperglycaemic conditions on brain GAGs/PGs are few and far between. We, therefore, looked into the changes in brain GAGs and PGs at various developmental stages of pre- and post-natal rats from non-diabetic and diabetic mothers as well as in adult rats induced with diabetes using a diabetogenic agent, Streptozotocin. Increased expression of GAGs especially that of heparan sulphate class in various developmental stages were observed in the brain as a result of in-utero hyperglycaemic condition but not in that of adult rats. Changes in disaccharides of heparan sulphate (HS) were observed in various developmental stages. Furthermore, various HSPGs namely, syndecans-1 and -3 and glypican-1 were overexpressed in offspring from diabetic mother. However, in adult diabetic rats, only glypican-1 was overexpressed. The offsprings from diabetic mothers became hyperphagic at the end of 8 weeks after birth which can have implications in the long run. Our results highlight the likely impact of the in-utero exposure of foetus to hyperglycaemic condition on brain GAGs/PGs compared to diabetic adult rats.

Chondroitin-4-sulfation negatively regulates axonal guidance and growth

Science.gov (United States)

Wang, Hang; Katagiri, Yasuhiro; McCann, Thomas E.; Unsworth, Edward; Goldsmith, Paul; Yu, Zu-Xi; Tan, Fei; Santiago, Lizzie; Mills, Edward M.; Wang, Yu; Symes, Aviva J.; Geller, Herbert M.

2008-01-01

Summary Glycosaminoglycan (GAG) side chains endow extracellular matrix proteoglycans with diversity and complexity based upon the length, composition, and charge distribution of the polysaccharide chain. Using cultured primary neurons, we show that specific sulfation in the GAG chains of chondroitin sulfate (CS) mediates neuronal guidance cues and axonal growth inhibition. Chondroitin-4-sulfate (CS-A), but not chondroitin-6-sulfate (CS-C), exhibits a strong negative guidance cue to mouse cerebellar granule neurons. Enzymatic and gene-based manipulations of 4-sulfation in the GAG side chains alter their ability to direct growing axons. Furthermore, 4-sulfated CS GAG chains are rapidly and significantly increased in regions that do not support axonal regeneration proximal to spinal cord lesions in mice. Thus, our findings provide the evidence showing that specific sulfation along the carbohydrate backbone carries instructions to regulate neuronal function. PMID:18768934

Effect of chondroitin sulfate proteoglycans on neuronal cell adhesion, spreading and neurite growth in culture

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Jingyu Jin

2018-01-01

Full Text Available As one major component of extracellular matrix (ECM in the central nervous system, chondroitin sulfate proteoglycans (CSPGs have long been known as inhibitors enriched in the glial scar that prevent axon regeneration after injury. Although many studies have shown that CSPGs inhibited neurite outgrowth in vitro using different types of neurons, the mechanism by which CSPGs inhibit axonal growth remains poorly understood. Using cerebellar granule neuron (CGN culture, in this study, we evaluated the effects of different concentrations of both immobilized and soluble CSPGs on neuronal growth, including cell adhesion, spreading and neurite growth. Neurite length decreased while CSPGs concentration arised, meanwhile, a decrease in cell density accompanied by an increase in cell aggregates formation was observed. Soluble CSPGs also showed an inhibition on neurite outgrowth, but it required a higher concentration to induce cell aggregates formation than coated CSPGs. We also found that growth cone size was significantly reduced on CSPGs and neuronal cell spreading was restrained by CSPGs, attributing to an inhibition on lamellipodial extension. The effect of CSPGs on neuron adhesion was further evidenced by interference reflection microscopy (IRM which directly demonstrated that both CGNs and cerebral cortical neurons were more loosely adherent to a CSPG substrate. These data demonstrate that CSPGs have an effect on cell adhesion and spreading in addition to neurite outgrowth.

Chondroitin sulphate extracted from antler cartilage using high hydrostatic pressure and enzymatic hydrolysis

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Chong-Tai Kim

2014-12-01

Full Text Available Chondroitin sulphate (CS, a major glycosaminoglycan, is an essential component of the extracellular matrix in cartilaginous tissues. Wapiti velvet antlers are a rich source of these molecules. The purpose of the present study was to develop an effective isolation procedure of CS from fresh velvet antlers using a combination of high hydrostatic pressure (100 MPa and enzymatic hydrolysis (papain. High CS extractability (95.1 ± 2.5% of total uronic acid was obtained following incubation (4 h at 50 °C with papain at pH 6.0 in 100 MPa compared to low extractability (19 ± 1.1% in ambient pressure (0.1 MPa. Antler CS fractions were isolated by Sephacryl S-300 chromatography and identified by western blot using an anti-CS monoclonal antibody. The antler CS fraction did not aggregate with hyaluronic acid in CL-2B chromatography and possessed DPPH radical scavenging activity at 78.3 ± 1.5%. The results indicated that high hydrostatic pressure and enzymatic hydrolysis procedure may be a useful tool for the isolation of CS from antler cartilaginous tissues.

Extracellular matrix in canine mammary tumors with special focus on versican, a versatile extracellular proteoglycan

NARCIS (Netherlands)

Erdélyi, Ildikó

2006-01-01

The extracellular matrix (ECM) research has become fundamental to understand cancer. This thesis focuses on the exploration of ECM composition and organization in canine mammary tumors, with a special interest in the large chondroitin-sulfate proteoglycan (PG), versican. Chapter 1 gives an

Human skin basement membrane-associated heparan sulphate proteoglycan: distinctive differences in ultrastructural localization as a function of developmental age

DEFF Research Database (Denmark)

Horiguchi, Y; Fine, J D; Couchman, J R

1991-01-01

was identical to that observed in neonatal and adult human skin. These findings demonstrate that active remodelling of the dermo-epidermal junction occurs during at least the first two trimesters, and affects not only basement membrane-associated structures but also specific antigens.......Recent studies have demonstrated that skin basement membrane components are expressed within the dermo-epidermal junction in an orderly sequence during human foetal development. We have investigated the ultrastructural localization of basement membrane-related antigens in human foetal skin...... at different developmental ages using two monoclonal antibodies to a well-characterized basement membrane-associated heparan sulphate proteoglycan. A series of foetal skin specimens (range, 54-142 gestational days) were examined using an immunoperoxidase immunoelectron microscopic technique. In specimens...

The effect of divalent salt in chondroitin sulfate solutions

Energy Technology Data Exchange (ETDEWEB)

Aranghel, D., E-mail: daranghe@nipne.ro [Horia Hulubei National Institute of Physics and Nuclear Engineering, Reactorului 30, RO-077125, POB-MG6, Magurele-Bucharest, Romania, daranghe@nipne.ro (Romania); Extreme Light Intrastructure Nuclear Physics (ELI-NP), Reactorului 30,RO-077125, POB-MG6, Magurele-Bucharest (Romania); Badita, C. R. [Horia Hulubei National Institute of Physics and Nuclear Engineering, Reactorului 30, RO-077125, POB-MG6, Magurele-Bucharest, Romania, daranghe@nipne.ro (Romania); University of Bucharest, Faculty of Physics, Atomiştilor 405, CP MG - 11, RO – 077125, Bucharest-Magurele (Romania); Radulescu, A. [Forschungszentrum Jülich GmbH, Jülich Centre for Neutron Science, 85747 Garching (Germany); Moldovan, L.; Craciunescu, O. [National Institute R& D for Biological Sciences, Splaiul Independenţei 296, sector 6, cod 060031, C.P. 17-16, Bucharest (Romania); Balasoiu, M. [Horia Hulubei National Institute of Physics and Nuclear Engineering, Reactorului 30, RO-077125, POB-MG6, Magurele-Bucharest, Romania, daranghe@nipne.ro (Romania); Joint Institute for Nuclear Research, 141980 Dubna, Moscow region (Russian Federation)

2016-03-25

Chondroitin-4 sulfate (CS4) is the main glycosaminoglycan extracted from bovine trachea. CS4 play an important role in osteoarthritis treatment, anticoagulant activity, reduces the degradation of cartilage matrix components, reduces necrosis and apoptosis of chondrocytes and reduces the activity of collagenase. Chondroitin sulfate is also responsible for proteoglycans degradation. Chondroitin sulfate can bind calcium ions with different affinities, depending on their sulfation position. The purpose of this study was to determine the structural properties and the influence of Ca{sup 2+} cations. We carried out measurements on CS4 solutions and mixtures of liquid CS4 with Ca{sup 2+} by Small-Angle Neutron Scattering (SANS). CS4 have a mass fractal behavior and the addition of a salt (CaCl{sub 2}) in CS4 solutions generates the appearance of a correlation peak due to local ordering between adjacent chains with inter-chain distances between 483 Å and 233 Å for a calcium concentration of 0.01% w/w.

The effect of divalent salt in chondroitin sulfate solutions

Science.gov (United States)

Aranghel, D.; Badita, C. R.; Radulescu, A.; Moldovan, L.; Craciunescu, O.; Balasoiu, M.

2016-03-01

Chondroitin-4 sulfate (CS4) is the main glycosaminoglycan extracted from bovine trachea. CS4 play an important role in osteoarthritis treatment, anticoagulant activity, reduces the degradation of cartilage matrix components, reduces necrosis and apoptosis of chondrocytes and reduces the activity of collagenase. Chondroitin sulfate is also responsible for proteoglycans degradation. Chondroitin sulfate can bind calcium ions with different affinities, depending on their sulfation position. The purpose of this study was to determine the structural properties and the influence of Ca2+ cations. We carried out measurements on CS4 solutions and mixtures of liquid CS4 with Ca2+ by Small-Angle Neutron Scattering (SANS). CS4 have a mass fractal behavior and the addition of a salt (CaCl2) in CS4 solutions generates the appearance of a correlation peak due to local ordering between adjacent chains with inter-chain distances between 483 Å and 233 Å for a calcium concentration of 0.01% w/w.

The effect of divalent salt in chondroitin sulfate solutions

International Nuclear Information System (INIS)

Aranghel, D.; Badita, C. R.; Radulescu, A.; Moldovan, L.; Craciunescu, O.; Balasoiu, M.

2016-01-01

Chondroitin-4 sulfate (CS4) is the main glycosaminoglycan extracted from bovine trachea. CS4 play an important role in osteoarthritis treatment, anticoagulant activity, reduces the degradation of cartilage matrix components, reduces necrosis and apoptosis of chondrocytes and reduces the activity of collagenase. Chondroitin sulfate is also responsible for proteoglycans degradation. Chondroitin sulfate can bind calcium ions with different affinities, depending on their sulfation position. The purpose of this study was to determine the structural properties and the influence of Ca"2"+ cations. We carried out measurements on CS4 solutions and mixtures of liquid CS4 with Ca"2"+ by Small-Angle Neutron Scattering (SANS). CS4 have a mass fractal behavior and the addition of a salt (CaCl_2) in CS4 solutions generates the appearance of a correlation peak due to local ordering between adjacent chains with inter-chain distances between 483 Å and 233 Å for a calcium concentration of 0.01% w/w.

Mast Cells Produce a Unique Chondroitin Sulfate Epitope.

Science.gov (United States)

Farrugia, Brooke L; Whitelock, John M; O'Grady, Robert; Caterson, Bruce; Lord, Megan S

2016-02-01

The granules of mast cells contain a myriad of mediators that are stored and protected by the sulfated glycosaminoglycan (GAG) chains that decorate proteoglycans. Whereas heparin is the GAG predominantly associated with mast cells, mast cell proteoglycans are also decorated with heparan sulfate and chondroitin sulfate (CS). This study investigated a unique CS structure produced by mast cells that was detected with the antibody clone 2B6 in the absence of chondroitinase ABC digestion. Mast cells in rodent tissue sections were characterized using toluidine blue, Leder stain and the presence of mast cell tryptase. The novel CS epitope was identified in rodent tissue sections and localized to cells that were morphologically similar to cells chemically identified as mast cells. The rodent mast cell-like line RBL-2H3 was also shown to express the novel CS epitope. This epitope co-localized with multiple CS proteoglycans in both rodent tissue and RBL-2H3 cultured cells. These findings suggest that the novel CS epitope that decorates mast cell proteoglycans may play a role in the way these chains are structured in mast cells. © 2016 The Histochemical Society.

Sulphate in Pregnancy

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Paul A. Dawson

2015-03-01

Full Text Available Sulphate is an obligate nutrient for healthy growth and development. Sulphate conjugation (sulphonation of proteoglycans maintains the structure and function of tissues. Sulphonation also regulates the bioactivity of steroids, thyroid hormone, bile acids, catecholamines and cholecystokinin, and detoxifies certain xenobiotics and pharmacological drugs. In adults and children, sulphate is obtained from the diet and from the intracellular metabolism of sulphur-containing amino acids. Dietary sulphate intake can vary greatly and is dependent on the type of food consumed and source of drinking water. Once ingested, sulphate is absorbed into circulation where its level is maintained at approximately 300 μmol/L, making sulphate the fourth most abundant anion in plasma. In pregnant women, circulating sulphate concentrations increase by twofold with levels peaking in late gestation. This increased sulphataemia, which is mediated by up-regulation of sulphate reabsorption in the maternal kidneys, provides a reservoir of sulphate to meet the gestational needs of the developing foetus. The foetus has negligible capacity to generate sulphate and thereby, is completely reliant on sulphate supply from the maternal circulation. Maternal hyposulphataemia leads to foetal sulphate deficiency and late gestational foetal death in mice. In humans, reduced sulphonation capacity has been linked to skeletal dysplasias, ranging from the mildest form, multiple epiphyseal dysplasia, to achondrogenesis Type IB, which results in severe skeletal underdevelopment and death in utero or shortly after birth. Despite being essential for numerous cellular and metabolic functions, the nutrient sulphate is largely unappreciated in clinical settings. This article will review the physiological roles and regulation of sulphate during pregnancy, with a particular focus on animal models of disturbed sulphate homeostasis and links to human pathophysiology.

Biosynthesis and function of chondroitin sulfate.

Science.gov (United States)

Mikami, Tadahisa; Kitagawa, Hiroshi

2013-10-01

Chondroitin sulfate proteoglycans (CSPGs) are principal pericellular and extracellular components that form regulatory milieu involving numerous biological and pathophysiological phenomena. Diverse functions of CSPGs can be mainly attributed to structural variability of their polysaccharide moieties, chondroitin sulfate glycosaminoglycans (CS-GAG). Comprehensive understanding of the regulatory mechanisms for CS biosynthesis and its catabolic processes is required in order to understand those functions. Here, we focus on recent advances in the study of enzymatic regulatory pathways for CS biosynthesis including successive modification/degradation, distinct CS functions, and disease phenotypes that have been revealed by perturbation of the respective enzymes in vitro and in vivo. Fine-tuned machineries for CS production/degradation are crucial for the functional expression of CS chains in developmental and pathophysiological processes. Control of enzymes responsible for CS biosynthesis/catabolism is a potential target for therapeutic intervention for the CS-associated disorders. Copyright © 2013 Elsevier B.V. All rights reserved.

The effect of a slightly acidic somatomedin peptide (ILAs) on the sulphation of proteoglycans from articular and growth plate chondrocytes in culture

International Nuclear Information System (INIS)

Corvol, M.-T.; Dumontier, M.-F.; Rappaport, R.; Guyda, H.; Posner, B.I.

1978-01-01

Chondrocyte cultures were prepared from rabbit growth plate (GPC) and articular (ARC) chondrocytes. These two cell types have distinct morphological characteristics. The cells reached maximum numbers by days 10 and 21 for ARC and GPC, respectively. The proteoglycans (PG) contained in the cellular pool were extracted and purified by DEAE cellulose chromatography. The effect of a partially purified somatomedin peptide with insulin-like activity on [ 35 S]sulphate incorporation into PG was evaluated. In both ARC and GPC a significant stimulation of [ 35 S]sulphate uptake into PG subunits was obtained with 1 ng Eq./ml of somatomedin peptide. In order to obtain the same stimulatory effect with porcine insulin, a 1000-fold greater concentration was required. The electrophoretic patterns of the PG subunits on acrylamide-agarose electrophoresis were identical on control incubations and after stimulation with the somatomedin peptide. These data demonstrate in vitro biological activity of this peptide on well differentiated articular and epiphyseal growth plate chondrocytes in culture. These cultures appear to provide a sensitive biological assay for somatomedin peptides. (author)

Rat mesangial cells in vitro synthesize a spectrum of proteoglycan species including those of the basement membrane and interstitium

DEFF Research Database (Denmark)

Thomas, G J; Shewring, L; McCarthy, K J

1995-01-01

is localized in the mesangium but is not found in the pericapillary glomerular basement membrane (GBM). Further characterization of the proteoglycans synthesized by RMC in vitro revealed: (i) a second large CSPG, identified as versican; (ii) two small dermatan sulphate proteoglycans identified as biglycan...

Brain ageing changes proteoglycan sulfation, rendering perineuronal nets more inhibitory.

Science.gov (United States)

Foscarin, Simona; Raha-Chowdhury, Ruma; Fawcett, James W; Kwok, Jessica C F

2017-06-28

Chondroitin sulfate (CS) proteoglycans in perineuronal nets (PNNs) from the central nervous system (CNS) are involved in the control of plasticity and memory. Removing PNNs reactivates plasticity and restores memory in models of Alzheimer's disease and ageing. Their actions depend on the glycosaminoglycan (GAG) chains of CS proteoglycans, which are mainly sulfated in the 4 (C4S) or 6 (C6S) positions. While C4S is inhibitory, C6S is more permissive to axon growth, regeneration and plasticity. C6S decreases during critical period closure. We asked whether there is a late change in CS-GAG sulfation associated with memory loss in aged rats. Immunohistochemistry revealed a progressive increase in C4S and decrease in C6S from 3 to 18 months. GAGs extracted from brain PNNs showed a large reduction in C6S at 12 and 18 months, increasing the C4S/C6S ratio. There was no significant change in mRNA levels of the chondroitin sulfotransferases. PNN GAGs were more inhibitory to axon growth than those from the diffuse extracellular matrix. The 18-month PNN GAGs were more inhibitory than 3-month PNN GAGs. We suggest that the change in PNN GAG sulfation in aged brains renders the PNNs more inhibitory, which lead to a decrease in plasticity and adversely affect memory.

Chondroitin sulfate proteoglycans regulate the growth, differentiation and migration of multipotent neural precursor cells through the integrin signaling pathway

Directory of Open Access Journals (Sweden)

Lü He-Zuo

2009-10-01

Full Text Available Abstract Background Neural precursor cells (NPCs are defined by their ability to proliferate, self-renew, and retain the potential to differentiate into neurons and glia. Deciphering the factors that regulate their behaviors will greatly aid in their use as potential therapeutic agents or targets. Chondroitin sulfate proteoglycans (CSPGs are prominent components of the extracellular matrix (ECM in the central nervous system (CNS and are assumed to play important roles in controlling neuronal differentiation and development. Results In the present study, we demonstrated that CSPGs were constitutively expressed on the NPCs isolated from the E16 rat embryonic brain. When chondroitinase ABC was used to abolish the function of endogenous CSPGs on NPCs, it induced a series of biological responses including the proliferation, differentiation and migration of NPCs, indicating that CSPGs may play a critical role in NPC development and differentiation. Finally, we provided evidence suggesting that integrin signaling pathway may be involved in the effects of CSPGs on NPCs. Conclusion The present study investigating the influence and mechanisms of CSPGs on the differentiation and migration of NPCs should help us to understand the basic biology of NPCs during CNS development and provide new insights into developing new strategies for the treatment of the neurological disorders in the CNS.

On the roles and regulation of chondroitin sulfate and heparan sulfate in zebrafish pharyngeal cartilage morphogenesis

DEFF Research Database (Denmark)

Holmborn, Katarina; Habicher, Judith; Kasza, Zsolt

2012-01-01

The present study addresses the roles of heparan sulfate (HS) proteoglycans and chondroitin sulfate (CS) proteoglycans in the development of zebrafish pharyngeal cartilage structures. uxs1 and b3gat3 mutants, predicted to have impaired biosynthesis of both HS and CS because of defective formation...... levels of CS than control larvae, whereas morpholino-mediated suppression of csgalnact1/csgalnact2 resulted in increased HS biosynthesis. Thus, the balance of the Extl3 and Csgalnact1/Csgalnact2 proteins influences the HS/CS ratio. A characterization of the pharyngeal cartilage element morphologies...

Accumulation of neurocan, a brain chondroitin sulfate proteoglycan, in association with the retinal vasculature in RCS rats.

Science.gov (United States)

Zhang, Yiqin; Rauch, Uwe; Perez, Maria-Thereza R

2003-03-01

To examine whether and how the retinal distribution of the chondroitin sulfate proteoglycan neurocan is affected after photoreceptor cell loss and whether it correlates with the multiple secondary cellular changes that accompany the photoreceptor degeneration. Retinas from normal rats (Sprague-Dawley; postnatal days [P]0-P70), RCS rats with dystrophic retinas (P0-P300), RCS-rdy(+) congenic rats with nondystrophic retinas (P0-202), and rhodopsin mutant rats, P23H (P0-P257) and S334ter (P0-P220), were processed for immunohistochemistry using a polyclonal antibody to rat neurocan. The overall distribution of neurocan was similar in all retinas examined. Neurocan immunostaining was detected over the nerve fiber layer, the plexiform layers, the photoreceptor outer segments region, and the ciliary epithelium. With age, labeling throughout the plexiform layers decreased continuously. In RCS rats however, conspicuous labeling was also seen in association with retinal vessels, from P15 onward. Accumulation of neurocan in association with the retinal vasculature does not correlate with photoreceptor cell loss, because it was not observed in the rhodopsin mutant rats. During the earliest stages of the disease, accumulation of debris in the subretinal space in RCS rats may be sufficient per se to initiate a cascade of metabolic changes that result in accumulation of neurocan. With time, the neurocan accumulated perivascularly may, by interaction with other matrix molecules, modulate at least some of the vascular alterations observed in this animal model.

Conformational studies on five octasaccharides isolated from chondroitin sulfate using NMR spectroscopy and molecular modeling

NARCIS (Netherlands)

Blanchard, V.; Chevalier, F.; Imberty, A.; Leeflang, B.R.; Sugahara, K.; Kamerling, J.P.

2007-01-01

Chondroitin sulfate proteoglycans (CS-PG) are involved in the regulation of the central nervous system in vertebrates due to their presence on cell surfaces and in the extracellular matrix of tissues. The CS moieties are built up from repeating -4)GlcA(β 1-3)GalNAc(β 1- disaccharide units, partly

Nanostructured 3D constructs based on chitosan and chondroitin sulphate multilayers for cartilage tissue engineering.

Directory of Open Access Journals (Sweden)

Joana M Silva

Full Text Available Nanostructured three-dimensional constructs combining layer-by-layer technology (LbL and template leaching were processed and evaluated as possible support structures for cartilage tissue engineering. Multilayered constructs were formed by depositing the polyelectrolytes chitosan (CHT and chondroitin sulphate (CS on either bidimensional glass surfaces or 3D packet of paraffin spheres. 2D CHT/CS multi-layered constructs proved to support the attachment and proliferation of bovine chondrocytes (BCH. The technology was transposed to 3D level and CHT/CS multi-layered hierarchical scaffolds were retrieved after paraffin leaching. The obtained nanostructured 3D constructs had a high porosity and water uptake capacity of about 300%. Dynamical mechanical analysis (DMA showed the viscoelastic nature of the scaffolds. Cellular tests were performed with the culture of BCH and multipotent bone marrow derived stromal cells (hMSCs up to 21 days in chondrogenic differentiation media. Together with scanning electronic microscopy analysis, viability tests and DNA quantification, our results clearly showed that cells attached, proliferated and were metabolically active over the entire scaffold. Cartilaginous extracellular matrix (ECM formation was further assessed and results showed that GAG secretion occurred indicating the maintenance of the chondrogenic phenotype and the chondrogenic differentiation of hMSCs.

Nanostructured 3D constructs based on chitosan and chondroitin sulphate multilayers for cartilage tissue engineering.

Science.gov (United States)

Silva, Joana M; Georgi, Nicole; Costa, Rui; Sher, Praveen; Reis, Rui L; Van Blitterswijk, Clemens A; Karperien, Marcel; Mano, João F

2013-01-01

Nanostructured three-dimensional constructs combining layer-by-layer technology (LbL) and template leaching were processed and evaluated as possible support structures for cartilage tissue engineering. Multilayered constructs were formed by depositing the polyelectrolytes chitosan (CHT) and chondroitin sulphate (CS) on either bidimensional glass surfaces or 3D packet of paraffin spheres. 2D CHT/CS multi-layered constructs proved to support the attachment and proliferation of bovine chondrocytes (BCH). The technology was transposed to 3D level and CHT/CS multi-layered hierarchical scaffolds were retrieved after paraffin leaching. The obtained nanostructured 3D constructs had a high porosity and water uptake capacity of about 300%. Dynamical mechanical analysis (DMA) showed the viscoelastic nature of the scaffolds. Cellular tests were performed with the culture of BCH and multipotent bone marrow derived stromal cells (hMSCs) up to 21 days in chondrogenic differentiation media. Together with scanning electronic microscopy analysis, viability tests and DNA quantification, our results clearly showed that cells attached, proliferated and were metabolically active over the entire scaffold. Cartilaginous extracellular matrix (ECM) formation was further assessed and results showed that GAG secretion occurred indicating the maintenance of the chondrogenic phenotype and the chondrogenic differentiation of hMSCs.

Oncofetal chondroitin sulfate glycosaminoglycans are key players in integrin signaling and tumor cell motility

DEFF Research Database (Denmark)

Clausen, Thomas Mandel; Bento Ayres Pereira, Marina Maria; Al Nakouzi, Nader

2016-01-01

Many tumors express proteoglycans modified with oncofetal chondroitin sulfate glycosaminoglycan chains (ofCS), which are normally restricted to the placenta. However, the role of ofCS in cancer is largely unknown. The function of ofCS in cancer was analyzed using the recombinant ofCS-binding VAR2...... revealed an overrepresentation of proteins involved in cell motility and integrin signaling, such as integrin-β1 (ITGB1) and integrin-α4 (ITGA4). Saturating concentrations of rVAR2 inhibited downstream integrin signaling, which was mimicked by knockdown of the core chondroitin sulfate synthesis enzymes β-1......,3-glucuronyltransferase 1 (B3GAT1) and chondroitin sulfate N-acetylgalactosaminyltransferase 1 (CSGALNACT1). The ofCS modification was highly expressed in both human and murine metastatic lesions in situ and preincubation or early intravenous treatment of tumor cells with rVAR2 inhibited seeding and spreading of tumor...

In vivo turnover of the basement membrane and other heparan sulfate proteoglycans of rat glomerulus

International Nuclear Information System (INIS)

Beavan, L.A.; Davies, M.; Couchman, J.R.; Williams, M.A.; Mason, R.M.

1989-01-01

The metabolic turnover of rat glomerular proteoglycans in vivo was investigated. Newly synthesized proteoglycans were labeled during a 7-h period after injecting sodium [35S]sulfate intraperitoneally. At the end of the labeling period a chase dose of sodium sulfate was given. Subsequently at defined times (0-163 h) the kidneys were perfused in situ with 0.01% cetylpyridinium chloride in phosphate-buffered saline to maximize the recovery of 35S-proteoglycans. Glomeruli were isolated from the renal cortex and analyzed for 35S-proteoglycans by autoradiographic, biochemical, and immunochemical methods. Grain counting of autoradiographs revealed a complex turnover pattern of 35S-labeled macromolecules, commencing with a rapid phase followed by a slower phase. Biochemical analysis confirmed the biphasic pattern and showed that the total population of [35S]heparan sulfate proteoglycans had a metabolic half-life (t1/2) of 20 and 60 h in the early and late phases, respectively. Heparan sulfate proteoglycans accounted for 80% of total 35S-proteoglycans, the remainder being chondroitin/dermatan sulfate proteoglycans. Whole glomeruli were extracted with 4% 3-[(cholamidopropyl)dimethy-lammonio]-1-propanesulfonate-4 M guanidine hydrochloride, a procedure which solubilized greater than 95% of the 35S-labeled macromolecules. Of these 11-13% was immunoprecipitated by an antiserum against heparan sulfate proteoglycan which, in immunolocalization experiments, showed specificity for staining the basement membrane of rat glomeruli. Autoradiographic analysis showed that 18% of total radioactivity present at the end of the labeling period was associated with the glomerular basement membrane

Transforming growth factor β-induced expression of chondroitin sulfate proteoglycans is mediated through non-Smad signaling pathways.

Science.gov (United States)

Jahan, Naima; Hannila, Sari S

2015-01-01

The expression of chondroitin sulfate proteoglycans (CSPGs) by reactive astrocytes is a major factor contributing to glial scarring and regenerative failure after spinal cord injury, but the molecular mechanisms underlying CSPG expression remain largely undefined. One contributing factor is transforming growth factor β (TGFβ), which is upregulated after injury and has been shown to induce expression of CSPGs in vitro. TGFβ typically mediates its effects through the Smad2/3 signaling pathway, and it has been suggested that this pathway is responsible for CSPG expression. However, there is evidence that TGFβ can also activate non-Smad signaling pathways. In this study, we report that TGFβ-induced expression of three different CSPGs--neurocan, brevican, and aggrecan--is mediated through non-Smad signaling pathways. We observed significant increases in TGFβ-induced expression of neurocan, brevican, and aggrecan following siRNA knockdown of Smad2 or Smad4, which indicates that Smad signaling is not required for the expression of these CSPGs. In addition, we show that neurocan, aggrecan, and brevican levels are significantly reduced when TGFβ is administered in the presence of either the PI3K inhibitor LY294002 or the mTOR inhibitor rapamycin, but not the MEK1/2 inhibitor U0126. This suggests that TGFβ mediates this effect through non-Smad-dependent activation of the PI3K-Akt-mTOR signaling pathway, and targeting this pathway may therefore be an effective means of reducing CSPG expression in the injured CNS. Copyright © 2014 Elsevier Inc. All rights reserved.

Self-assembly of the hydrogel polymer chain consisting of chitosan and chondroitin sulphate in the presence of theophylline

International Nuclear Information System (INIS)

Lopes, Lais C.; Piai, Juliana F.; Fajardo, Andre R.; Rubira, Adley F.; Muniz, Edvani C.

2009-01-01

In this work, polyelectronic complex (PEC) consisting of two polysaccharides were developed. One is chitosan (QT), cationic polymer, produced by the chitin deacetylation and the other is chondroitin sulphate (CS), anionic polymer, extracted from bovine or porcine aorta. The PECs were prepared in the presence of theophylline (TEO) for evaluating the influence of this drug in the polymer chains reorganization, as well as, studying the mechanical properties and release of SC and TEO in aqueous solutions on different pH conditions. By the obtained results, it was observed that the 84QT/15SC/TEO (% in weight) hydrogel is pH responsive because the CS releasing is more effective at pH 8, while the release of the TEO is higher at pH 2. The hydrogel showed mechanical properties more resistant to pH 2, 8 and 10 and this was attributed to interactions between the polymer chains. Finally, the X-rays profile showed the presence of peaks associated to reorganization of the chains in the hydrogel is at times larger than the hydrogel in the absence of solute. (author)

Placental sequestration of Plasmodium falciparum malaria parasites is mediated by the interaction between VAR2CSA and chondroitin sulfate A on syndecan-1

DEFF Research Database (Denmark)

Ayres Pereira, Marina; Mandel Clausen, Thomas; Pehrson, Caroline

2016-01-01

During placental malaria, Plasmodium falciparum infected erythrocytes sequester in the placenta, causing health problems for both the mother and fetus. The specific adherence is mediated by the VAR2CSA protein, which binds to placental chondroitin sulfate (CS) on chondroitin sulfate proteoglycans......-down experiments using placental extracts from whole placenta or syncytiotrophoblast microvillous cell membranes showed three distinct CSPGs available for VAR2CSA adherence. Further examination of these three CSPGs by immunofluorescence and proximity ligation assays showed that syndecan-1 is the main receptor...... for VAR2CSA mediated placental adherence. We further show that the commonly used placental choriocarcinoma cell line, BeWo, express a different set of proteoglycans than those present on placental syncytiotrophoblast and may not be the most biologically relevant model to study placental malaria. Syncytial...

Decorin is one of the proteoglycans expressed in Walker 256 rat mammary carcinoma

Directory of Open Access Journals (Sweden)

S.M. Oba-Shinjo

2003-08-01

Full Text Available Proteoglycan and glycosaminoglycan content was analyzed in a model of rat mammary carcinoma to study the roles of these compounds in tumorigenesis. Hyaluronic acid and proteoglycans bearing chondroitin and/or dermatan sulfate chains were detected in solid tumors obtained after subcutaneous inoculation of Walker 256 rat carcinoma cells. About 10% of sulfated glycosaminoglycan chains corresponded to heparan sulfate. The small leucine-rich proteoglycan, decorin, was identified as one of the proteoglycans, in addition to others of higher molecular weight, by cross-reaction with an antiserum raised against pig laryngeal decorin and by N-terminal amino acid sequencing. Decorin was separated from other proteoglycans by hydrophobic chromatography and its complete structure was determined. It has a molecular weight of about 85 kDa and a dermatan chain of 45 kDa with 4-sulfated disaccharides. After degradation of the glycosaminoglycan chain, three core proteins of different molecular weight (36, 46 and 56 kDa were identified. The presence of hyaluronic acid and decorin has been reported in a variety of tumors and tumor cells. In the Walker 256 mammary carcinoma model, hyaluronic acid may play an important role in tumor progression, since it provides a more hydrated extracellular matrix. On the other hand, decorin, which is expressed by stromal cells, represents a host defense response to tumor growth.

Glycomics expression analysis of sulfated glycosaminoglycans of human colorectal cancer tissues and non-neoplastic mucosa by electrospray ionization mass spectrometry.

Science.gov (United States)

Marolla, Ana Paula Cleto; Waisberg, Jaques; Saba, Gabriela Tognini; Waisberg, Daniel Reis; Margeotto, Fernando Beani; Pinhal, Maria Aparecida da Silva

2015-01-01

To determine the presence of glycosaminoglycans in the extracellular matrix of connective tissue from neoplastic and non-neoplastic colorectal tissues, since it has a central role in tumor development and progression. Tissue samples from neoplastic and non-neoplastic colorectal tissues were obtained from 64 operated patients who had colorectal carcinoma with no distant metastases. Expressions of heparan sulphate, chondroitin sulphate, dermatan sulphate and their fragments were analyzed by electrospray ionization mass spectrometry, with the technique for extraction and quantification of glycosaminoglycans after proteolysis and electrophoresis. The statistical analysis included mean, standard deviation, and Student'st test. The glycosaminoglycans extracted from colorectal tissue showed three electrophoretic bands in agarose gel. Electrospray ionization mass spectrometry showed characteristic disaccharide fragments from glycosaminoglycans, indicating their structural characterization in the tissues analyzed. Some peaks in the electrospray ionization mass spectrometry were not characterized as fragments of sugars, indicating the presence of fragments of the protein structure of proteoglycans generated during the glycosaminoglycan purification. The average amount of chondroitin and dermatan increased in the neoplastic tissue compared to normal tissue (p=0.01). On the other hand, the average amount of heparan decreased in the neoplastic tissue compared to normal tissue (p= 0.03). The method allowed the determination of the glycosaminoglycans structural profile in colorectal tissue from neoplastic and non-neoplastic colorectal tissue. Neoplastic tissues showed greater amounts of chondroitin sulphate and dermatan sulphate compared to non-neoplastic tissues, while heparan sulphate was decreased in neoplastic tissues.

Glycomics expression analysis of sulfated glycosaminoglycans of human colorectal cancer tissues and non-neoplastic mucosa by electrospray ionization mass spectrometry

Energy Technology Data Exchange (ETDEWEB)

Marolla, Ana Paula Cleto [Universidade Federal de São Paulo, São Paulo, SP (Brazil); Waisberg, Jaques [Hospital do Servidor Público Estadual, São Paulo, SP (Brazil); Faculdade de Medicina do ABC, Santo André, SP (Brazil); Saba, Gabriela Tognini [Faculdade de Medicina do ABC, Santo André, SP (Brazil); Waisberg, Daniel Reis [Faculdade de Medicina da Universidade de São Paulo, São Paulo, SP (Brazil); Margeotto, Fernando Beani; Pinhal, Maria Aparecida da Silva [Faculdade de Medicina do ABC, Santo André, SP (Brazil)

2015-07-01

To determine the presence of glycosaminoglycans in the extracellular matrix of connective tissue from neoplastic and non-neoplastic colorectal tissues, since it has a central role in tumor development and progression. Tissue samples from neoplastic and non-neoplastic colorectal tissues were obtained from 64 operated patients who had colorectal carcinoma with no distant metastases. Expressions of heparan sulphate, chondroitin sulphate, dermatan sulphate and their fragments were analyzed by electrospray ionization mass spectrometry, with the technique for extraction and quantification of glycosaminoglycans after proteolysis and electrophoresis. The statistical analysis included mean, standard deviation, and Student’s t test. The glycosaminoglycans extracted from colorectal tissue showed three electrophoretic bands in agarose gel. Electrospray ionization mass spectrometry showed characteristic disaccharide fragments from glycosaminoglycans, indicating their structural characterization in the tissues analyzed. Some peaks in the electrospray ionization mass spectrometry were not characterized as fragments of sugars, indicating the presence of fragments of the protein structure of proteoglycans generated during the glycosaminoglycan purification. The average amount of chondroitin and dermatan increased in the neoplastic tissue compared to normal tissue (p=0.01). On the other hand, the average amount of heparan decreased in the neoplastic tissue compared to normal tissue (p= 0.03). The method allowed the determination of the glycosaminoglycans structural profile in colorectal tissue from neoplastic and non-neoplastic colorectal tissue. Neoplastic tissues showed greater amounts of chondroitin sulphate and dermatan sulphate compared to non-neoplastic tissues, while heparan sulphate was decreased in neoplastic tissues.

Glycomics expression analysis of sulfated glycosaminoglycans of human colorectal cancer tissues and non-neoplastic mucosa by electrospray ionization mass spectrometry

International Nuclear Information System (INIS)

Marolla, Ana Paula Cleto; Waisberg, Jaques; Saba, Gabriela Tognini; Waisberg, Daniel Reis; Margeotto, Fernando Beani; Pinhal, Maria Aparecida da Silva

2015-01-01

To determine the presence of glycosaminoglycans in the extracellular matrix of connective tissue from neoplastic and non-neoplastic colorectal tissues, since it has a central role in tumor development and progression. Tissue samples from neoplastic and non-neoplastic colorectal tissues were obtained from 64 operated patients who had colorectal carcinoma with no distant metastases. Expressions of heparan sulphate, chondroitin sulphate, dermatan sulphate and their fragments were analyzed by electrospray ionization mass spectrometry, with the technique for extraction and quantification of glycosaminoglycans after proteolysis and electrophoresis. The statistical analysis included mean, standard deviation, and Student’s t test. The glycosaminoglycans extracted from colorectal tissue showed three electrophoretic bands in agarose gel. Electrospray ionization mass spectrometry showed characteristic disaccharide fragments from glycosaminoglycans, indicating their structural characterization in the tissues analyzed. Some peaks in the electrospray ionization mass spectrometry were not characterized as fragments of sugars, indicating the presence of fragments of the protein structure of proteoglycans generated during the glycosaminoglycan purification. The average amount of chondroitin and dermatan increased in the neoplastic tissue compared to normal tissue (p=0.01). On the other hand, the average amount of heparan decreased in the neoplastic tissue compared to normal tissue (p= 0.03). The method allowed the determination of the glycosaminoglycans structural profile in colorectal tissue from neoplastic and non-neoplastic colorectal tissue. Neoplastic tissues showed greater amounts of chondroitin sulphate and dermatan sulphate compared to non-neoplastic tissues, while heparan sulphate was decreased in neoplastic tissues

Biphasic role of chondroitin sulfate in cardiac differentiation of embryonic stem cells through inhibition of Wnt/β-catenin signaling.

Directory of Open Access Journals (Sweden)

Robert D Prinz

Full Text Available The glycosaminoglycan chondroitin sulfate is a critical component of proteoglycans on the cell surface and in the extracellular matrix. As such, chondroitin sulfate side chains and the sulfation balance of chondroitin play important roles in the control of signaling pathways, and have a functional importance in human disease. In contrast, very little is known about the roles of chondroitin sulfate molecules and sulfation patterns during mammalian development and cell lineage specification. Here, we report a novel biphasic role of chondroitin sulfate in the specification of the cardiac cell lineage during embryonic stem cell differentiation through modulation of Wnt/beta-catenin signaling. Lineage marker analysis demonstrates that enzymatic elimination of endogenous chondroitin sulfates leads to defects specifically in cardiac differentiation. This is accompanied by a reduction in the number of beating cardiac foci. Mechanistically, we show that endogenous chondroitin sulfate controls cardiac differentiation in a temporal biphasic manner through inhibition of the Wnt/beta-catenin pathway, a known regulatory pathway for the cardiac lineage. Treatment with a specific exogenous chondroitin sulfate, CS-E, could mimic these biphasic effects on cardiac differentiation and Wnt/beta-catenin signaling. These results establish chondroitin sulfate and its sulfation balance as important regulators of cardiac cell lineage decisions through control of the Wnt/beta-catenin pathway. Our work suggests that targeting the chondroitin biosynthesis and sulfation machinery is a novel promising avenue in regenerative strategies after heart injury.

Biphasic role of chondroitin sulfate in cardiac differentiation of embryonic stem cells through inhibition of Wnt/β-catenin signaling.

Science.gov (United States)

Prinz, Robert D; Willis, Catherine M; van Kuppevelt, Toin H; Klüppel, Michael

2014-01-01

The glycosaminoglycan chondroitin sulfate is a critical component of proteoglycans on the cell surface and in the extracellular matrix. As such, chondroitin sulfate side chains and the sulfation balance of chondroitin play important roles in the control of signaling pathways, and have a functional importance in human disease. In contrast, very little is known about the roles of chondroitin sulfate molecules and sulfation patterns during mammalian development and cell lineage specification. Here, we report a novel biphasic role of chondroitin sulfate in the specification of the cardiac cell lineage during embryonic stem cell differentiation through modulation of Wnt/beta-catenin signaling. Lineage marker analysis demonstrates that enzymatic elimination of endogenous chondroitin sulfates leads to defects specifically in cardiac differentiation. This is accompanied by a reduction in the number of beating cardiac foci. Mechanistically, we show that endogenous chondroitin sulfate controls cardiac differentiation in a temporal biphasic manner through inhibition of the Wnt/beta-catenin pathway, a known regulatory pathway for the cardiac lineage. Treatment with a specific exogenous chondroitin sulfate, CS-E, could mimic these biphasic effects on cardiac differentiation and Wnt/beta-catenin signaling. These results establish chondroitin sulfate and its sulfation balance as important regulators of cardiac cell lineage decisions through control of the Wnt/beta-catenin pathway. Our work suggests that targeting the chondroitin biosynthesis and sulfation machinery is a novel promising avenue in regenerative strategies after heart injury.

Molecular interactions between chondroitin-dermatan sulfate and growth factors/receptors/matrix proteins.

Science.gov (United States)

Mizumoto, Shuji; Yamada, Shuhei; Sugahara, Kazuyuki

2015-10-01

Recent functional studies on chondroitin sulfate-dermatan sulfate (CS-DS) demonstrated its indispensable roles in various biological events including brain development and cancer. CS-DS proteoglycans exert their physiological activity through interactions with specific proteins including growth factors, cell surface receptors, and matrix proteins. The characterization of these interactions is essential for regulating the biological functions of CS-DS proteoglycans. Although amino acid sequences on the bioactive proteins required for these interactions have already been elucidated, the specific saccharide sequences involved in the binding of CS-DS to target proteins have not yet been sufficiently identified. In this review, recent findings are described on the interaction between CS-DS and some proteins which are especially involved in the central nervous system and cancer development/metastasis. Copyright © 2015. Published by Elsevier Ltd.

Specificity of the human proteoglycan radioimmunoassay

International Nuclear Information System (INIS)

Gysen, P.; Heynen, G.; Franchimont, P.

1981-01-01

The human articular cartilagineous proteoglycans (PG) R.I.A. is highly specific. The PG used as the standard and the 125 I labelled molecule appear to be pure. Under these conditions, all the potential interfering substances which have been tested show no cross reaction. For instance, the Ag-Ab equilibrium is not affected by adding human IgG, human albumin, hyaluronic acid, chondroitin sulfate, rat type II collagen or total human serum proteins. This R.I.A. also exhibits a species spcificity since there is no cross reaction with rat PG and negligible cross section with dog PG. The results obtained after addition of enzymes to the antigen demonstrate that the antigenic sites are localized on the protein region and not on the glycosaminoglycan region of the molecule [fr

Specificity of the human proteoglycan radioimmunoassay

Energy Technology Data Exchange (ETDEWEB)

Gysen, P.; Heynen, G.; Franchimont, P. (Liege Univ. (Belgium))

1981-01-01

The human articular cartilagineous proteoglycans (PG) R.I.A. is highly specific. The PG used as the standard and the /sup 125/I labelled molecule appear to be pure. Under these conditions, all the potential interfering substances which have been tested show no cross reaction. For instance, the Ag-Ab equilibrium is not affected by adding human IgG, human albumin, hyaluronic acid, chondroitin sulfate, rat type II collagen or total human serum proteins. This R.I.A. also exhibits a species spcificity since there is no cross reaction with rat PG and negligible cross section with dog PG. The results obtained after addition of enzymes to the antigen demonstrate that the antigenic sites are localized on the protein region and not on the glycosaminoglycan region of the molecule.

Low buoyant density proteoglycans from saline and dissociative extracts of embryonic chicken retinas

Energy Technology Data Exchange (ETDEWEB)

Morris, J.E.; Ting, Y.P.; Birkholz-Lambrecht, A.

1984-03-01

Retinas were labeled in culture with (/sup 3/H)glucosamine or (/sup 3/H)leucine and (/sup 35/S)sulfate and extracted sequentially with physiologically balanced saline and 4 M guanidine HCl. They were dialyzed into associative conditions (0.5 M NaCl) and chromatographed on agarose columns. Under these conditions, some of the proteoglycans were associated in massive complexes that showed low buoyant densities when centrifuged in CsCl density gradients under dissociative conditions (4 M guanidine HCl). Much of the label in these complexes was in molecules other than proteoglycans. Most of the proteoglycans, however, were included on the agarose columns, where they appeared to be constitutionally of low buoyant density. They resisted attempts to separate potential low buoyant density contaminants from the major proteoglycans by direct CsCl density gradient centrifugation or by the fractionation of saline or 8 M urea extracts on diethylaminoethyl-Sephacel. The diethylaminoethyl-Sephacel fractions were either subjected to CsCl density gradient centrifugation or were chromatographed on Sephacryl S-300, in both cases before and after alkaline cleavage, to confirm the presence of typical O-linked glycosaminoglycans. The medium and balanced salt extracts were enriched in chondroitin sulfate and other sulfated macromolecules, possibly highly sulfated oligosaccharides, that resisted digestion by chondroitinase ABC but were electrophoretically less mobile than heparan sulfate. Guanidine HCl or urea extracts of the residues were mixtures of high and low density proteoglycans that were enriched in heparan sulfate.

Molecular dissection of placental malaria protein VAR2CSA interaction with a chemo-enzymatically synthesized chondroitin sulfate library

DEFF Research Database (Denmark)

Sugiura, Nobuo; Clausen, Thomas Mandel; Shioiri, Tatsuasa

2016-01-01

with chondroitin sulfate (CS) proteoglycans present in the placental tissue. CS is a linear acidic polysaccharide composed of repeating disaccharide units of d-glucuronic acid and N-acetyl-d-galactosamine that are modified by sulfate groups at different positions. Previous reports have shown that placental......-adhering IEs were associated with an unusually low sulfated form of chondroitin sulfate A (CSA) and that a partially sulfated dodecasaccharide is the minimal motif for the interaction. However, the fine molecular structure of this CS chain remains unclear. In this study, we have characterized the CS chain...... that interacts with a recombinant minimal CS-binding region of VAR2CSA (rVAR2) using a CS library of various defined lengths and sulfate compositions. The CS library was chemo-enzymatically synthesized with bacterial chondroitin polymerase and recombinant CS sulfotransferases. We found that C-4 sulfation...

Medical Gains of Chondroitin Sulfate Upon Fucosylation.

Science.gov (United States)

Pomin, Vitor H

2015-01-01

Chondroitin sulfate (CS) is a glycosaminoglycan (GAG) composed of alternating N-acetyl galactosamine and glucuronic acid units within disaccharide building blocks. CS is a key functional component in proteoglycans of cartilaginous tissues. Owing to its numerous biological roles, CS is widely explored in the pharmaceutical market as nutraceutical ingredient commonly utilized against arthritis, osteoarthrosis, and sometimes osteoporosis. Tissues like shark cartilage and bovine trachea are common sources of CS. Nonetheless, a new CS type has been introduced and investigated in the last few decades in what regards its medical potentials. It is named fucosylated chondroitin sulfate (FucCS). This less common CS type is isolated exclusively from the body wall of sea cucumbers. The presence of fucosyl branching units in the holothurian FucCS gives to this unique GAG, therapeutic properties in various pathophysiological systems which are inexistent in the common CS explored in the market. Examples of these systems are coagulation, thrombosis, hemodialysis, atherosclerosis, cellular growth, angiogenesis, fibrosis, tumor growth, inflammation, viral and protozoan infections, hyperglycemia, diabetes-related pathological events and tissue damage. This report aims at describing the medical benefits gained upon fucosylation of CS. Clinical prospects of these medical benefits are also discussed herein.

Ice as a Green-Structure-Directing Agent in the Synthesis of Macroporous MWCNTs and Chondroitin Sulphate Composites

Science.gov (United States)

Nardecchia, Stefania; Serrano, María Concepción; García-Argüelles, Sara; Maia Da Costa, Marcelo E. H.; Ferrer, María Luisa; Gutiérrez, María C.

2017-01-01

The incorporation of multi-walled carbon nanotubes (MWCNTs) into chondroitin sulphate-based scaffolds and the effect on the structural, mechanical, conductive, and thermal properties of the resulting scaffolds is investigated. Three-dimensional hierarchical materials are prepared upon the application of the ice segregation-induced self-assembly (ISISA) process. The use of ice as structure-directing agents avoids chemicals typically used for this purpose (e.g., surfactants, block copolymers, etc.), hence, emphasising the green features of this soft-templating approach. We determine the critical parameters that control the morphology of the scaffolds formed upon ice-templating (i.e., MWCNTs type, freezing conditions, polymer and MWCNT concentration). MWCNTs are surface functionalized by acidic treatment. MWCNT functionalization is characterized by Raman, Fourier transfer infrared (FTIR) and X-ray Photoelectron (XPS) spectroscopies. Scanning electron microscopy (SEM) analysis and porosity studies reveal that MWCNT content modifies the morphology of the macroporous structure, which decreases by increasing MWCNT concentration. Differences in scaffold morphology should be translated into their conductivity and mechanical properties. As a general trend, the Young’s modulus and the electrical conductivity of the scaffolds increase with the MWCNT content. Preliminary biocompatibility tests with human osteoblast-like cells also reveal the capability of these structures to support cell growth. PMID:28772715

Symptom and structure modification in osteoarthritis with pharmaceutical-grade chondroitin sulfate: what's the evidence?

Science.gov (United States)

Hochberg, M; Chevalier, X; Henrotin, Y; Hunter, D J; Uebelhart, D

2013-03-01

Osteoarthritis is a chronic disease characterized by irreversible damage to joint structures, including loss of articular cartilage, osteophyte formation, alterations in the subchondral bone and synovial inflammation. It has been shown that chondroitin sulfate interferes with the progression of structural changes in joint tissues and is used in the management of patients with osteoarthritis. This review summarizes data from relevant reports describing the mechanisms of action of chondroitin sulfate that may explain the beneficial effects of the drug and examines the evidence for clinical efficacy of oral chondroitin sulfate in osteoarthritis. Data included in the review were derived from a literature search in PubMed. Literature searches were performed in PubMed using the search terms 'chondroitin sulfate', 'pharmaceutical-grade', 'osteoarthritis', 'randomized clinical trials', 'humans'. The MEDLINE database was searched from January 1996 through August 2012 for all randomized controlled trials, meta-analyses, systematic reviews, and review articles of chondroitin sulfate in osteoarthritis. Chondroitin sulfate exerts in vitro a beneficial effect on the metabolism of different cell lines: chondrocytes, synoviocytes and cells from subchondral bone, all involved in osteoarthritis. It increases type II collagen and proteoglycan synthesis in human articular chondrocytes and is able to reduce the production of some pro-inflammatory factors and proteases, to reduce the cellular death process, and improve the anabolic/catabolic balance of the extracellular cartilage matrix (ECM). Clinical trials have reported a beneficial effect of chondroitin sulfate on pain and function. The structure-modifying effects of chondroitin sulfate have been reported and analyzed in recent meta-analyses. The results in knee osteoarthritis demonstrate a small but significant reduction in the rate of decline in joint space width. Because chondroitin sulfate quality of several nutraceuticals has

Semi-synthesis of chondroitin sulfate-E from chondroitin sulfate-A

OpenAIRE

Cai, Chao; Solakyildirim, Kemal; Yang, Bo; Beaudet, Julie M.; Weyer, Amanda; Linhardt, Robert J.; Zhang, Fuming

2012-01-01

Chondroitin sulfate-E (chondroitin-4, 6-disulfate) was prepared from chondroitin sulfate-A (chondroitin-4 - sulfate) by regioselective sulfonation, performed using trimethylamine sulfur trioxide in formamide under argon. The structure of semi-synthetic chondroitin sulfate-E was analyzed by PAGE, 1H NMR, 13C NMR, 2D NMR and disaccharide analysis and compared with natural chondroitin sulfate-E. Both semi-synthetic and natural chondroitin sulfate-E were each biotinylated and immobilized on BIAco...

Oncofetal Chondroitin Sulfate Glycosaminoglycans are Key Players in Integrin Signaling and Tumor Cell Motility

Science.gov (United States)

Clausen, Thomas Mandel; Pereira, Marina Ayres; Al Nakouzi, Nader; Oo, Htoo Zarni; Agerbæk, Mette Ø; Lee, Sherry; Ørum-Madsen, Maj Sofie; Christensen, Anders Riis; El-Naggar, Amal; Grandgenett, Paul M.; Grem, Jean L.; Hollingsworth, Michael A.; Holst, Peter J.; Theander, Thor; Sorensen, Poul H.; Daugaard, Mads; Salanti, Ali

2016-01-01

Many tumors express proteoglycans modified with oncofetal chondroitin sulfate glycosaminoglycan chains (ofCS), which are normally restricted to the placenta. However, the role of ofCS in cancer is largely unknown. The function of ofCS in cancer was analyzed using the recombinant ofCS-binding VAR2CSA protein (rVAR2) derived from the malaria parasite, Plasmodium falciparum. We demonstrate that ofCS plays a key role in tumor cell motility by affecting canonical integrin signaling pathways. Binding of rVAR2 to tumor cells inhibited the interaction of cells with extracellular matrix (ECM) components, which correlated with decreased phosphorylation of Src kinase. Moreover, rVAR2 binding decreased migration, invasion and anchorage-independent growth of tumor cells in vitro. Mass spectrometry of ofCS-modified proteoglycan complexes affinity purified from tumor cell lines on rVAR2 columns, revealed an overrepresentation of proteins involved in cell motility and integrin signaling, such as integrin β1 (ITGB1) and integrin α4 (ITGA4). Saturating concentrations of rVAR2 inhibited downstream integrin signaling, which was mimicked by knockdown of the core CS synthesis enzymes Beta-1,3-Glucuronyltransferase 1 (B3GAT1) and Chondroitin Sulfate N-Acetylgalactosaminyltransferase 1 (CSGALNACT1). The ofCS modification was highly expressed in both human and murine metastatic lesions in situ and pre-incubation or early intravenous treatment of tumor cells with rVAR2 inhibited seeding and spreading of tumor cells in mice. This was associated with a significant increase in survival of the animals. These data functionally link ofCS modifications with cancer cell motility and further highlights ofCS as a novel therapeutic cancer target. Implications The cancer specific expression of oncofetal chondroitin sulfate aids in metastatic phenotypes and is a candidate target for therapy. PMID:27655130

The Effect of Chondroitin Sulphate and Hyaluronic Acid on Chondrocytes Cultured within a Fibrin-Alginate Hydrogel

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Christopher J. Little

2014-09-01

Full Text Available Osteoarthritis is a painful degenerative joint disease that could be better managed if tissue engineers can develop methods to create long-term engineered articular cartilage tissue substitutes. Many of the tissue engineered cartilage constructs currently available lack the chemical stimuli and cell-friendly environment that promote the matrix accumulation and cell proliferation needed for use in joint cartilage repair. The goal of this research was to test the efficacy of using a fibrin-alginate hydrogel containing hyaluronic acid (HA and/or chondroitin sulphate (CS supplements for chondrocyte culture. Neonatal porcine chondrocytes cultured in fibrin-alginate hydrogels retained their phenotype better than chondrocytes cultured in monolayer, as evidenced by analysis of their relative expression of type II versus type I collagen mRNA transcripts. HA or CS supplementation of the hydrogels increased matrix glycosaminoglycan (GAG production during the first week of culture. However, the effects of these supplements on matrix accumulation were not additive and were no longer observed after two weeks of culture. Supplementation of the hydrogels with CS or a combination of both CS and HA increased the chondrocyte cell population after two weeks of culture. Statistical analysis indicated that the HA and CS treatment effects on chondrocyte numbers may be additive. This research suggests that supplementation with CS and/or HA has positive effects on cartilage matrix production and chondrocyte proliferation in three-dimensional (3D fibrin-alginate hydrogels.

Acute Exacerbations of COPD Are Associated With Increased Expression of Heparan Sulfate and Chondroitin Sulfate in BAL.

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Papakonstantinou, Eleni; Klagas, Ioannis; Roth, Michael; Tamm, Michael; Stolz, Daiana

2016-03-01

Acute exacerbations of COPD (AECOPDs) are associated with accelerated aggravation of clinical symptoms and deterioration of pulmonary function. The mechanisms by which exacerbations may contribute to airway remodeling and declined lung function are poorly understood. We investigated whether AECOPDs are associated with differential expression of glycosaminoglycans in BAL in a cohort of 97 patients with COPD. Patients with COPD with either stable disease (n = 53) or AECOPD (n = 44) and undergoing diagnostic bronchoscopy were matched for demographics and lung function parameters. Levels of heparan sulfate, chondroitin sulfate, dermatan sulfate, and matrix metalloproteinases (MMPs) in BAL were measured by enzyme-linked immunosorbent assay. Heparan sulfate and chondroitin sulfate were significantly increased in BAL of patients during exacerbations. Levels of heparan sulfate were higher in the BAL of patients with microbial infections. Chondroitin sulfate was negatively correlated with FEV1 % predicted but not with diffusing capacity of lung for carbon monoxide % predicted, indicating that chondroitin sulfate is associated with airway remodeling, leading to obstruction rather than to emphysema. Furthermore, heparan sulfate and chondroitin sulfate were significantly correlated with MMP-9, MMP-2, and MMP-12 in BAL, indicating that they were cleaved from their respective proteoglycans by MMPs and subsequently washed out in BAL. During AECOPD, there is increased expression of heparan sulfate and chondroitin sulfate in BAL. These molecules are significantly correlated with MMPs in BAL, indicating that they may be associated with airway remodeling and may lead to lung function decline during exacerbations of COPD. Copyright © 2016 American College of Chest Physicians. Published by Elsevier Inc. All rights reserved.

Glycosaminoglycans and Proteoglycans

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Vitor H. Pomin

2018-02-01

Full Text Available In this editorial to MDPI Pharmaceuticals special issue “Glycosaminoglycans and Proteoglycans” we describe in outline the common structural features of glycosaminoglycans and the characteristics of proteoglycans, including the intracellular proteoglycan, serglycin, cell-surface proteoglycans, like syndecans and glypicans, and the extracellular matrix proteoglycans, like aggrecan, perlecan, and small leucine-rich proteoglycans. The context in which the pharmaceutical uses of glycosaminoglycans and proteoglycans are presented in this special issue is given at the very end.

The spatiotemporal relationships between chondroitin sulfate proteoglycans and terminations of calcitonin gene related peptide and parvalbumin immunoreactive afferents in the spinal cord of mouse embryos.

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Wang, Liqing; Yu, Chao; Wang, Jun; Zhao, Hui; Chan, Sun-On

2017-08-10

Chondroitin sulfate (CS) proteoglycans (PGs) are a family of complex molecules in the extracellular matrix and cell surface that regulate axon growth and guidance during development of the central nervous system. In this study, the expression of CSPGs was investigated in the mouse spinal cord at late embryonic and neonatal stages using CS-56 antibody. CS immunoreactivity was observed abundantly in ventral regions of spinal cord of embryonic day (E) 15 embryos. At E16 to E18, CS expression spread dorsally, but never reached the superficial layers of the dorsal horn. This pattern was maintained until postnatal day 4, the latest stage examined. Antibodies against calcitonin gene related peptide (CGRP) and parvalbumin (PV) were employed to label primary afferents from nociceptors and proprioceptors, respectively. CGRP-immunoreactive fibers terminated in the superficial regions of the dorsal horn where CSPGs were weakly expressed, whereas PV-immunoreactive fibers were found in CSPG-rich regions in the ventral horn. Therefore, we conclude that CS expression is spatiotemporally regulated in the spinal cord, which correlates to the termination of sensory afferents. This pattern suggests a role of CSPGs on patterning afferents in the spinal cord, probably through a differential response of axons to these growth inhibitory molecules. Copyright © 2017 Elsevier B.V. All rights reserved.

Iduronic acid in chondroitin/dermatan sulfate affects directional migration of aortic smooth muscle cells.

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Barbara Bartolini

Full Text Available Aortic smooth muscle cells produce chondroitin/dermatan sulfate (CS/DS proteoglycans that regulate extracellular matrix organization and cell behavior in normal and pathological conditions. A unique feature of CS/DS proteoglycans is the presence of iduronic acid (IdoA, catalyzed by two DS epimerases. Functional ablation of DS-epi1, the main epimerase in these cells, resulted in a major reduction of IdoA both on cell surface and in secreted CS/DS proteoglycans. Downregulation of IdoA led to delayed ability to re-populate wounded areas due to loss of directional persistence of migration. DS-epi1-/- aortic smooth muscle cells, however, had not lost the general property of migration showing even increased speed of movement compared to wild type cells. Where the cell membrane adheres to the substratum, stress fibers were denser whereas focal adhesion sites were fewer. Total cellular expression of focal adhesion kinase (FAK and phospho-FAK (pFAK was decreased in mutant cells compared to control cells. As many pathological conditions are dependent on migration, modulation of IdoA content may point to therapeutic strategies for diseases such as cancer and atherosclerosis.

Proteoglycan from salmon nasal cartridge promotes in vitro wound healing of fibroblast monolayers via the CD44 receptor

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Ito, Gen; Kobayashi, Takeshi; Takeda, Yoshie; Sokabe, Masahiro

2015-01-01

Highlights: • Proteoglycan from salmon nasal cartridge (SNC-PG) promoted wound healing in fibroblast monolayers. • SNC-PG stimulated both cell proliferation and cell migration. • Interaction between chondroitin sulfate-units and CD44 is responsible for the effect. - Abstract: Proteoglycans (PGs) are involved in various cellular functions including cell growth, adhesion, and differentiation; however, their physiological roles are not fully understood. In this study, we examined the effect of PG purified from salmon nasal cartilage (SNC-PG) on wound closure using tissue-cultured cell monolayers, an in vitro wound-healing assay. The results indicated that SNC-PG significantly promoted wound closure in NIH/3T3 cell monolayers by stimulating both cell proliferation and cell migration. SNC-PG was effective in concentrations from 0.1 to 10 μg/ml, but showed much less effect at higher concentrations (100–1000 μg/ml). The effect of SNC-PG was abolished by chondroitinase ABC, indicating that chondroitin sulfates (CSs), a major component of glycosaminoglycans (GAGs) in SNC-PG, are crucial for the SNC-PG effect. Furthermore, chondroitin 6-sulfate (C-6-S), a major CS of SNC-PG GAGs, could partially reproduce the SNC-PG effect and partially inhibit the binding of SNC-PG to cells, suggesting that SNC-PG exerts its effect through an interaction between the GAGs in SNC-PG and the cell surface. Neutralization by anti-CD44 antibodies or CD44 knockdown abolished SNC-PG binding to the cells and the SNC-PG effect on wound closure. These results suggest that interactions between CS-rich GAG-chains of SNC-PG and CD44 on the cell surface are responsible for the SNC-PG effect on wound closure

Proteoglycan from salmon nasal cartridge promotes in vitro wound healing of fibroblast monolayers via the CD44 receptor

Energy Technology Data Exchange (ETDEWEB)

Ito, Gen; Kobayashi, Takeshi; Takeda, Yoshie [Department of Physiology, Nagoya University Graduate School of Medicine, Nagoya, Aichi 466-8550 (Japan); Sokabe, Masahiro, E-mail: msokabe@med.nagoya-u.ac.jp [Department of Physiology, Nagoya University Graduate School of Medicine, Nagoya, Aichi 466-8550 (Japan); Mechanobiology Laboratory, Nagoya University Graduate School of Medicine, Nagoya, Aichi 466-8550 (Japan); Mechanobiology Institute Singapore, National University of Singapore, Singapore 117411 (Singapore)

2015-01-16

Highlights: • Proteoglycan from salmon nasal cartridge (SNC-PG) promoted wound healing in fibroblast monolayers. • SNC-PG stimulated both cell proliferation and cell migration. • Interaction between chondroitin sulfate-units and CD44 is responsible for the effect. - Abstract: Proteoglycans (PGs) are involved in various cellular functions including cell growth, adhesion, and differentiation; however, their physiological roles are not fully understood. In this study, we examined the effect of PG purified from salmon nasal cartilage (SNC-PG) on wound closure using tissue-cultured cell monolayers, an in vitro wound-healing assay. The results indicated that SNC-PG significantly promoted wound closure in NIH/3T3 cell monolayers by stimulating both cell proliferation and cell migration. SNC-PG was effective in concentrations from 0.1 to 10 μg/ml, but showed much less effect at higher concentrations (100–1000 μg/ml). The effect of SNC-PG was abolished by chondroitinase ABC, indicating that chondroitin sulfates (CSs), a major component of glycosaminoglycans (GAGs) in SNC-PG, are crucial for the SNC-PG effect. Furthermore, chondroitin 6-sulfate (C-6-S), a major CS of SNC-PG GAGs, could partially reproduce the SNC-PG effect and partially inhibit the binding of SNC-PG to cells, suggesting that SNC-PG exerts its effect through an interaction between the GAGs in SNC-PG and the cell surface. Neutralization by anti-CD44 antibodies or CD44 knockdown abolished SNC-PG binding to the cells and the SNC-PG effect on wound closure. These results suggest that interactions between CS-rich GAG-chains of SNC-PG and CD44 on the cell surface are responsible for the SNC-PG effect on wound closure.

Clinical efficacy and safety of glucosamine, chondroitin sulphate, their combination, celecoxib or placebo taken to treat osteoarthritis of the knee: 2-year results from GAIT.

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Sawitzke, Allen D; Shi, Helen; Finco, Martha F; Dunlop, Dorothy D; Harris, Crystal L; Singer, Nora G; Bradley, John D; Silver, David; Jackson, Christopher G; Lane, Nancy E; Oddis, Chester V; Wolfe, Fred; Lisse, Jeffrey; Furst, Daniel E; Bingham, Clifton O; Reda, Domenic J; Moskowitz, Roland W; Williams, H James; Clegg, Daniel O

2010-08-01

Knee osteoarthritis (OA) is a major cause of pain and functional limitation in older adults, yet longer-term studies of medical treatment of OA are limited. To evaluate the efficacy and safety of glucosamine and chondroitin sulphate (CS), alone or in combination, as well as celecoxib and placebo on painful knee OA over 2 years. A 24-month, double-blind, placebo-controlled study, conducted at nine sites in the US ancillary to the Glucosamine/chondroitin Arthritis Intervention Trial, enrolled 662 patients with knee OA who satisfied radiographic criteria (Kellgren/Lawrence grade 2 or 3 changes and baseline joint space width of at least 2 mm). This subset continued to receive their randomised treatment: glucosamine 500 mg three times daily, CS 400 mg three times daily, the combination of glucosamine and CS, celecoxib 200 mg daily, or placebo over 24 months. The primary outcome was a 20% reduction in Western Ontario and McMaster University Osteoarthritis Index (WOMAC) pain over 24 months. Secondary outcomes included an Outcome Measures in Rheumatology/Osteoarthritis Research Society International response and change from baseline in WOMAC pain and function. Compared with placebo, the odds of achieving a 20% reduction in WOMAC pain were celecoxib: 1.21, glucosamine: 1.16, combination glucosamine/CS: 0.83 and CS alone: 0.69, and were not statistically significant. Over 2 years, no treatment achieved a clinically important difference in WOMAC pain or function as compared with placebo. However, glucosamine and celecoxib showed beneficial but not significant trends. Adverse reactions were similar among treatment groups and serious adverse events were rare for all treatments.

Oncofetal Chondroitin Sulfate Glycosaminoglycans Are Key Players in Integrin Signaling and Tumor Cell Motility.

Science.gov (United States)

Clausen, Thomas Mandel; Pereira, Marina Ayres; Al Nakouzi, Nader; Oo, Htoo Zarni; Agerbæk, Mette Ø; Lee, Sherry; Ørum-Madsen, Maj Sofie; Kristensen, Anders Riis; El-Naggar, Amal; Grandgenett, Paul M; Grem, Jean L; Hollingsworth, Michael A; Holst, Peter J; Theander, Thor; Sorensen, Poul H; Daugaard, Mads; Salanti, Ali

2016-12-01

Many tumors express proteoglycans modified with oncofetal chondroitin sulfate glycosaminoglycan chains (ofCS), which are normally restricted to the placenta. However, the role of ofCS in cancer is largely unknown. The function of ofCS in cancer was analyzed using the recombinant ofCS-binding VAR2CSA protein (rVAR2) derived from the malaria parasite, Plasmodium falciparum We demonstrate that ofCS plays a key role in tumor cell motility by affecting canonical integrin signaling pathways. Binding of rVAR2 to tumor cells inhibited the interaction of cells with extracellular matrix (ECM) components, which correlated with decreased phosphorylation of Src kinase. Moreover, rVAR2 binding decreased migration, invasion, and anchorage-independent growth of tumor cells in vitro Mass spectrometry of ofCS-modified proteoglycan complexes affinity purified from tumor cell lines on rVAR2 columns revealed an overrepresentation of proteins involved in cell motility and integrin signaling, such as integrin-β1 (ITGB1) and integrin-α4 (ITGA4). Saturating concentrations of rVAR2 inhibited downstream integrin signaling, which was mimicked by knockdown of the core chondroitin sulfate synthesis enzymes β-1,3-glucuronyltransferase 1 (B3GAT1) and chondroitin sulfate N-acetylgalactosaminyltransferase 1 (CSGALNACT1). The ofCS modification was highly expressed in both human and murine metastatic lesions in situ and preincubation or early intravenous treatment of tumor cells with rVAR2 inhibited seeding and spreading of tumor cells in mice. This was associated with a significant increase in survival of the animals. These data functionally link ofCS modifications with cancer cell motility and further highlights ofCS as a novel therapeutic cancer target. The cancer-specific expression of ofCS aids in metastatic phenotypes and is a candidate target for therapy. Mol Cancer Res; 14(12); 1288-99. ©2016 AACR. ©2016 American Association for Cancer Research.

Specific genes involved in synthesis and editing of heparan sulfate proteoglycans show altered expression patterns in breast cancer

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Fernández-Vega, Iván; García, Olivia; Crespo, Ainara; Castañón, Sonia; Menéndez, Primitiva; Astudillo, Aurora; Quirós, Luis M

2013-01-01

The expression of a specific set of genes controls the different structures of heparan sulfate proteoglycans (HSPGs), which are involved in the growth, invasion and metastatic properties of cancerous cells. The purpose of this study is to increase knowledge of HSPG alterations in breast cancer. Twenty-three infiltrating ductal adenocarcinomas (IDCs), both metastatic and non-metastatic were studied. A transcriptomic approach to the structure of heparan sulfate (HS) chains was used, employing qPCR to analyze both the expression of the enzymes involved in their biosynthesis and editing, as well as the proteoglycan core proteins. Since some of these proteoglycans can also carry chondroitin sulfate chains, we extended the study to include the genes involved in the biosynthesis of these glycosaminoglycans. Histochemical techniques were also used to analyze tissular expression of particular genes showing significant expression differences, of potential interest. No significant change in transcription was detected in approximately 70% of analyzed genes. However, 13 demonstrated changes in both tumor types (40% showing more intense deregulation in the metastatic), while 5 genes showed changes only in non-metastatic tumors. Changes were related to 3 core proteins: overexpression of syndecan-1 and underexpression of glypican-3 and perlecan. HS synthesis was affected by lower levels of some 3-O-sulfotransferase transcripts, the expression of NDST4 and, only in non metastatic tumors, higher levels of extracellular sulfatases. Furthermore, the expression of chondroitin sulfate also was considerably affected, involving both the synthesis of the saccharidic chains and sulfations at all locations. However, the pro-metastatic enzyme heparanase did not exhibit significant changes in mRNA expression, although in metastatic tumors it appeared related to increased levels of the most stable form of mRNA. Finally, the expression of heparanase 2, which displays anti-metastatic features

Two faces of chondroitin sulfate proteoglycan in spinal cord repair: a role in microglia/macrophage activation.

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Asya Rolls

2008-08-01

Full Text Available BACKGROUND: Chondroitin sulfate proteoglycan (CSPG is a major component of the glial scar. It is considered to be a major obstacle for central nervous system (CNS recovery after injury, especially in light of its well-known activity in limiting axonal growth. Therefore, its degradation has become a key therapeutic goal in the field of CNS regeneration. Yet, the abundant de novo synthesis of CSPG in response to CNS injury is puzzling. This apparent dichotomy led us to hypothesize that CSPG plays a beneficial role in the repair process, which might have been previously overlooked because of nonoptimal regulation of its levels. This hypothesis is tested in the present study. METHODS AND FINDINGS: We inflicted spinal cord injury in adult mice and examined the effects of CSPG on the recovery process. We used xyloside to inhibit CSPG formation at different time points after the injury and analyzed the phenotype acquired by the microglia/macrophages in the lesion site. To distinguish between the resident microglia and infiltrating monocytes, we used chimeric mice whose bone marrow-derived myeloid cells expressed GFP. We found that CSPG plays a key role during the acute recovery stage after spinal cord injury in mice. Inhibition of CSPG synthesis immediately after injury impaired functional motor recovery and increased tissue loss. Using the chimeric mice we found that the immediate inhibition of CSPG production caused a dramatic effect on the spatial organization of the infiltrating myeloid cells around the lesion site, decreased insulin-like growth factor 1 (IGF-1 production by microglia/macrophages, and increased tumor necrosis factor alpha (TNF-alpha levels. In contrast, delayed inhibition, allowing CSPG synthesis during the first 2 d following injury, with subsequent inhibition, improved recovery. Using in vitro studies, we showed that CSPG directly activated microglia/macrophages via the CD44 receptor and modulated neurotrophic factor secretion by

Inhibition of chondroitin sulfate glycosaminoglycans incorporation affected odontoblast differentiation in cultured embryonic mouse molars.

Science.gov (United States)

Liu, Lipei; Chen, Weiting; Li, Lefeng; Xu, Fangfang; Jiang, Beizhan

2017-12-01

Chondroitin sulfate proteoglycan (CSPG) is an important component of extracellular matrix (ECM), it is composed of a core protein and one or more chondroitin sulfate glycosaminoglycan side chains (CS-GAGs). To investigate the roles of its CS-GAGs in dentinogenesis, the mouse mandibular first molar tooth germs at early bell stage were cultivated with or without β-xyloside. As expected, the CS-GAGs were inhibited on their incorporation to CSPGs by β-xyloside, accompanied by the change of morphology of the cultured tooth germs. The histological results and the transmission electron microscopy (TEM) investigation indicated that β-xyloside exhibited obvious inhibiting effects on odontoblasts differentiation compared with the control group. Meanwhile the results of immunohistochemistry, in situ hybridization and quantitative RT-PCR for type I collagen, dentin matrix acidic phosphoprotein 1 and dentin sialophosphoprotein, the products of differentiated odontoblasts, further proved that odontoblasts differentiation was inhibited. Collagen fibers detected in TEM decreased and arranged in disorder as well. Thus we conclude that the inhibition of CS-GAGs incorporation to CSPGs can affect odontoblast differentiation in cultured embryonic mouse molars.

Ultrastructural localization of the core protein of a basement membrane-specific chondroitin sulfate proteoglycan in adult rat skin

DEFF Research Database (Denmark)

McCarthy, K J; Horiguchi, Y; Couchman, J R

1990-01-01

Basement membranes are complex extracellular matrices present at epithelial/mesenchymal interfaces of tissues. The dermal-epidermal junction has been shown to contain numerous components, some of the most well known being laminin, types IV and VII collagens, heparan sulfate proteoglycan, fibronec...

High cell density cultivation of Escherichia coli K4 in a microfiltration bioreactor: a step towards improvement of chondroitin precursor production

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De Rosa Mario

2011-02-01

Full Text Available Abstract Background The bacteria Escherichia coli K4 produces a capsular polysaccharide (K4 CPS whose backbone is similar to the non sulphated chondroitin chain. The chondroitin sulphate is one of the major components of the extra-cellular matrix of the vertebrate connective tissues and a high value molecule, widely employed as active principle in the treatment of osteoarthritis. It is usually obtained by extraction from animal tissues, but the risk of virus contaminations, as well as the scarceness of raw material, makes this productive process unsafe and unable to satisfy the growing market demand. In previous studies a new biotechnological process to produce chondroitin from Escherichia coli K4 capsular polysaccharide was investigated and a 1.4 g·L-1 K4 CPS concentration was reached using fed-batch fermentation techniques. In this work, on the trail of these results, we exploited new fermentation strategies to further improve the capsular polysaccharide production. Results The inhibitory effect of acetate on the bacterial cells growth and K4 CPS production was studied in shake flask conditions, while a new approach, that combined the optimization of the feeding profiles, the improvement of aeration conditions and the use of a microfiltration bioreactor, was investigated in three different types of fermentation processes. High polysaccharide concentrations (4.73 ± 0.2 g·L-1, with corresponding average yields (0.13 ± 0.006 gK4 CPS·gcdw-1, were obtained; the increase of K4 CPS titre, compared to batch and fed-batch results, was of 16-fold and 3.3-fold respectively, while average yield was almost 3.5 and 1.4 fold higher. Conclusion The increase of capsular polysaccharide titre confirmed the validity of the proposed fermentation strategy and opened the way to the use of the microfiltration bioreactor for the biotechnological production of chondroitin.

Extracellular matrix of smooth muscle cells: interaction of collagen type V with heparan sulfate proteoglycan

International Nuclear Information System (INIS)

Gay, S.; Hoeoek, M.; Gay, R.E.; Magargal, W.W.; Reynertson, R.H.

1986-01-01

Alteration in the extracellular matrix produced by smooth muscle cells may play a role in the development of atherosclerotic lesions. Consequently the authors have initiated studies on the structural organization of the extracellular matrix produced by cultured smooth muscle cells. Immunohisotological examination of this matrix using well-characterized mono- and polyclonal antibodies showed a partial codistribution of heparan sulfate (HS) proteoglycans with a number of different matrix components including collagen types I, III, IV, V and VI, laminin and fibronectin. Subsequent binding studies between isolated matrix proteins and HS showed that the polysaccharide interacts strongly with type V collagen and to a lesser extent with fibronectin as well as collagen types III and VI. The interaction between type V and HS was readily inhibited by heparin and highly sulfated HS but not be dermatan sulfate, chondroitin sulfate or HS with a low sulfate content. Furthermore, [ 35 S]-HS proteoglycans isolated from cultured smooth muscle cells could be adsorbed on a column of sepharose conjugated with native type V collagen and eluted in a salt gradient. Hence, the interaction between type V and HS may play a major part in stabilizing the extracellular matrix of the vessel wall

Chondroitin 6-O-sulfate ameliorates experimental autoimmune encephalomyelitis.

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Miyamoto, Katsuichi; Tanaka, Noriko; Moriguchi, Kota; Ueno, Rino; Kadomatsu, Kenji; Kitagawa, Hiroshi; Kusunoki, Susumu

2014-05-01

Chondroitin sulfate proteoglycans (CSPGs) are the main component of the extracellular matrix in the central nervous system (CNS) and influence neuroplasticity. Although CSPG is considered an inhibitory factor for nerve repair in spinal cord injury, it is unclear whether CSPG influences the pathogenetic mechanisms of neuroimmunological diseases. We induced experimental autoimmune encephalomyelitis (EAE) in chondroitin 6-O-sulfate transferase 1-deficient (C6st1(-/-)) mice. C6ST1 is the enzyme that transfers sulfate residues to position 6 of N-acetylgalactosamine in the sugar chain of CSPG. The phenotypes of EAE in C6st1(-/-) mice were more severe than those in wild-type (WT) mice were. In adoptive-transfer EAE, in which antigen-reactive T cells from WT mice were transferred to C6st1(-/-) and WT mice, phenotypes were significantly more severe in C6st1(-/-) than in WT mice. The recall response of antigen-reactive T cells was not significantly different among the groups. Furthermore, the number of pathogenic T cells within the CNS was also not considerably different. When EAE was induced in C6ST1 transgenic mice with C6ST1 overexpression, the mice showed considerably milder symptoms compared with those in WT mice. In conclusion, the presence of sulfate at position 6 of N-acetylgalactosamine of CSPG may influence the effecter phase of EAE to prevent the progression of pathogenesis. Thus, modification of the carbohydrate residue of CSPG may be a novel therapeutic strategy for neuroimmunological diseases such as multiple sclerosis.

The Compact and Biologically Relevant Structure of Inter-α-inhibitor Is Maintained by the Chondroitin Sulfate Chain and Divalent Cations.

Science.gov (United States)

Scavenius, Carsten; Nikolajsen, Camilla Lund; Stenvang, Marcel; Thøgersen, Ida B; Wyrożemski, Łukasz; Wisniewski, Hans-Georg; Otzen, Daniel E; Sanggaard, Kristian W; Enghild, Jan J

2016-02-26

Inter-α-inhibitor is a proteoglycan of unique structure. The protein consists of three subunits, heavy chain 1, heavy chain 2, and bikunin covalently joined by a chondroitin sulfate chain originating at Ser-10 of bikunin. Inter-α-inhibitor interacts with an inflammation-associated protein, tumor necrosis factor-inducible gene 6 protein, in the extracellular matrix. This interaction leads to transfer of the heavy chains from the chondroitin sulfate of inter-α-inhibitor to hyaluronan and consequently to matrix stabilization. Divalent cations and heavy chain 2 are essential co-factors in this transfer reaction. In the present study, we have investigated how divalent cations in concert with the chondroitin sulfate chain influence the structure and stability of inter-α-inhibitor. The results showed that Mg(2+) or Mn(2+), but not Ca(2+), induced a conformational change in inter-α-inhibitor as evidenced by a decrease in the Stokes radius and a bikunin chondroitin sulfate-dependent increase of the thermodynamic stability. This structure was shown to be essential for the ability of inter-α-inhibitor to participate in extracellular matrix stabilization. In addition, the data revealed that bikunin was positioned adjacent to both heavy chains and that the two heavy chains also were in close proximity. The chondroitin sulfate chain interacted with all protein components and inter-α-inhibitor dissociated when it was degraded. Conventional purification protocols result in the removal of the Mg(2+) found in plasma and because divalent cations influence the conformation and affect function it is important to consider this when characterizing the biological activity of inter-α-inhibitor. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

NG2 proteoglycan increases mesangial cell proliferation and extracellular matrix production

International Nuclear Information System (INIS)

Xiong Jing; Wang Yang; Zhu, Zhonghua; Liu Jianshe; Wang Yumei; Zhang Chun; Hammes, Hans-Peter; Lang, Florian; Feng Yuxi

2007-01-01

As a membrane-spanning protein, NG2 chondroitin sulfate proteoglycan interacts with molecules on both sides of plasma membrane. The present study explored the role of NG2 in the pathogenesis of diabetic nephropathy. In the normal kidneys, NG2 was observed predominantly in glomerular mesangium, Bowman's capsule and interstitial vessels. Both mRNA and protein expression in kidneys was significantly higher in strepozotocin-induced diabetic rats than that in normal rats. In the cultured rat mesangial cell line HBZY-1, overexpression of NG2 promoted mesangial cell proliferation and extracellular matrix (ECM) production, such as type VI collagen and laminin. Furthermore, target knockdown of NG2 resulted in decreased cell proliferation and ECM formation. The observations suggest that NG2 is up-regulated in diabetic nephropathy. It actively participates in the development and progression of glomerulosclerosis by stimulating proliferation of mesangial cells and deposition of ECM

Chondroitin for osteoarthritis

Science.gov (United States)

Singh, Jasvinder A.; Noorbaloochi, Shahrzad; MacDonald, Roderick; Maxwell, Lara J.

2016-01-01

Background Osteoarthritis, a common joint disorder, is one of the leading causes of disability. Chondroitin has emerged as a new treatment. Previous meta-analyses have shown contradictory results on the efficacy of chondroitin. This, in addition to the publication of more trials, necessitates a systematic review. Objectives To evaluate the benefit and harm of oral chondroitin for treating osteoarthritis compared with placebo or a comparator oral medication including, but not limited to, nonsteroidal anti-inflammatory drugs (NSAIDs), analgesics, opioids, and glucosamine or other “herbal” medications. Search methods We searched seven databases up to November 2013, including the Cochrane Central Register of Controlled Trials (CENTRAL), Ovid MEDLINE, CINAHL, EMBASE, Science Citation Index (Web of Science) and Current Controlled Trials. We searched the US Food and Drug Administration (FDA) and European Medicines Agency (EMEA) websites for adverse effects. Trial registers were not searched. Selection criteria All randomized or quasi-randomized clinical trials lasting longer than two weeks, studying adults with osteoarthritis in any joint, and comparing chondroitin with placebo, an active control such as NSAIDs, or other “herbal” supplements such as glucosamine. Data collection and analysis Two review authors independently performed all title assessments, data extractions, and risk of bias assessments. Main results Forty-three randomized controlled trials including 4,962 participants treated with chondroitin and 4,148 participants given placebo or another control were included. The majority of trials were in knee OA, with few in hip and hand OA. Trial duration varied from 1 month to 3 years. Participants treated with chondroitin achieved statistically significantly and clinically meaningful better pain scores (0–100) in studies less than 6 months than those given placebo with an absolute risk difference of 10% lower (95% confidence interval (CI), 15% to 6% lower

Placental Sequestration of Plasmodium falciparum Malaria Parasites Is Mediated by the Interaction Between VAR2CSA and Chondroitin Sulfate A on Syndecan-1

Science.gov (United States)

Mao, Yang; Resende, Mafalda; Daugaard, Mads; Riis Kristensen, Anders; Damm, Peter; G. Theander, Thor; R. Hansson, Stefan; Salanti, Ali

2016-01-01

During placental malaria, Plasmodium falciparum infected erythrocytes sequester in the placenta, causing health problems for both the mother and fetus. The specific adherence is mediated by the VAR2CSA protein, which binds to placental chondroitin sulfate (CS) on chondroitin sulfate proteoglycans (CSPGs) in the placental syncytium. However, the identity of the CSPG core protein and the cellular impact of the interaction have remain elusive. In this study we identified the specific CSPG core protein to which the CS is attached, and characterized its exact placental location. VAR2CSA pull-down experiments using placental extracts from whole placenta or syncytiotrophoblast microvillous cell membranes showed three distinct CSPGs available for VAR2CSA adherence. Further examination of these three CSPGs by immunofluorescence and proximity ligation assays showed that syndecan-1 is the main receptor for VAR2CSA mediated placental adherence. We further show that the commonly used placental choriocarcinoma cell line, BeWo, express a different set of proteoglycans than those present on placental syncytiotrophoblast and may not be the most biologically relevant model to study placental malaria. Syncytial fusion of the BeWo cells, triggered by forskolin treatment, caused an increased expression of placental CS-modified syndecan-1. In line with this, we show that rVAR2 binding to placental CS impairs syndecan-1-related Src signaling in forskolin treated BeWo cells, but not in untreated cells. PMID:27556547

Effect of chondroitin sulphate on synovitis of knee osteoarthritic patients.

Science.gov (United States)

Tío, Laura; Orellana, Cristobal; Pérez-García, Selene; Piqueras, Laura; Escudero, Paula; Juarranz, Yasmina; Garcia-Giralt, Natalia; Montañés, Francisco; Farran, Aina; Benito, Pere; Gomariz, Rosa P; Sanz, María-Jesús; Monfort, Jordi

2017-07-07

To evaluate by ultrasonography the effect of chondroitin sulfate (CS) on synovitis in patients with knee osteoarthritis (KOA). To collaborate in the understanding of the biochemical mechanisms involved in the synovial inflammation process. Randomized, single-blind, controlled trial involving 70 patients with primary KOA treated for 6 months with CS or acetaminophen (ACT). Evaluation of KOA status at baseline, 6 weeks, 3 and 6 months included: ultrasonography to assess synovitis (following the OMERACT expertise group definition), visual analogue scale and Lequesne index to measure pain and function, and ELISA to quantify inflammatory mediators in serum and synovial fluid. Synovitis presence was reduced by 50% in the CS group while a 123% increase was observed in ACT group. Conversely, patients without initial synovitis and treated with ACT reached 85.71% synovitis onset, but only 25% in CS group. Both therapies improved articular function, but only CS resulted in significant pain improvement at the end of the treatment. Changes in RANTES and UCN synovial fluid concentration were associated with CS treatment. Treatment with CS had a sustained beneficial effect, preventing synovitis onset or reducing its presence as well as reducing KOA symptoms. ACT ameliorated clinical symptoms but had no effect on inflammation. The CS anti-inflammatory effect could be related to the observed changes in RANTES and UCN concentration. Copyright © 2017 Elsevier España, S.L.U. All rights reserved.

Use of chondroitin sulphate and glucosamine sulphate in degenerative changes in TMJ: a systematic review Utilização de sulfato de condroitina e sulfato de glicosamina nas alterações degenerativas da ATM: uma revisão sistematica

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Eduardo Machado

2012-08-01

Full Text Available INTRODUCTION: Degenerative changes in Temporomandibular Joint (TMJ have increased in prevalence and severity over the years. Within this context, it's necessary to obtain safe and effective therapies for control and management of the patient in cases of osteoarthritis and osteoarthrosis of the TMJ. Therapeutic options range from intra-articular infiltration protocols, occlusal splints, pharmacological therapies and physiotherapy and educational measures. The alternative treatment with structure-modifying agents, like as chondroitin and glucosamine sulphates, showed promising results, and especially safety. Thus, through a systematic literature review, this study aimed to analyze and discuss effectiveness and safety of chondroitin and glucosamine in degenerative changes of the TMJ. METHODS: Survey in research bases MEDLINE, Cochrane, EMBASE, Pubmed, Lilacs and BBO, between the years of 1966 and January 2009, with focus in randomized clinical trial (RCTs and quasi-randomized clinical trials, systematic reviews and meta-analysis. RESULTS: After application of the inclusion criteria 2 articles were selected, both randomized controlled double-blind clinical trials, which evaluated the effectiveness of chondroitin and glucosamine in degenerative changes of the TMJ. CONCLUSIONS: There is the necessity of further RCT, with representative samples and long follow-up time, to obtainment more precise cause-effect relationships and to achieve an effective and objective protocol involving chondroitin and glucosamine in cases of degenerative changes of the TMJ.INTRODUÇÃO: as alterações degenerativas da Articulação Temporomandibular (ATM têm aumentado em prevalência e em severidade ao longo dos anos. Dentro desse contexto, surge a necessidade de se obter terapêuticas efetivas e seguras para o controle e o manejo do paciente em situações de osteoartrite e osteoartrose da ATM. As opções terapêuticas variam desde protocolos de infiltra

Differentiation of EL4 lymphoma cells by tumoral environment is associated with inappropriate expression of the large chondroitin sulfate proteoglycan PG-M and the tumor-associated antigen HTgp-175.

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Rottiers, P; Verfaillie, T; Contreras, R; Revets, H; Desmedt, M; Dooms, H; Fiers, W; Grooten, J

1998-11-09

Progression to malignancy of transformed cells involves complex genetic alterations and aberrant gene expression patterns. While aberrant gene expression is often caused by alterations in individual genes, the contribution of the tumoral environment to the triggering of this gene expression is less well established. The stable but heterogeneous expression in cultured EL4/13 cells of a novel tumor-associated antigen, designated as HTgp-175, was chosen for the investigation of gene expression during tumor formation. Homogeneously HTgp-175-negative EL4/13 cells, isolated by cell sorting or obtained by subcloning, acquired HTgp-175 expression as a result of tumor formation. The tumorigenicity of HTgp-175-negative vs. HTgp-175-positive EL4 variants was identical, indicating that induction but not selection accounted for the phenotypic switch from HTgp-175-negative to HTgp-175-positive. Although mutagenesis experiments showed that the protein was not essential for tumor establishment, tumor-derived cells showed increased malignancy, linking HTgp-175 expression with genetic changes accompanying tumor progression. This novel gene expression was not an isolated event, since it was accompanied by ectopic expression of the large chondroitin sulfate proteoglycan PG-M and of normal differentiation antigens. We conclude that signals derived from the tumoral microenvironment contribute significantly to the aberrant gene expression pattern of malignant cells, apparently by fortuitous activation of differentiation processes and cause expression of novel differentiation antigens as well as of inappropriate tumor-associated and ectopic antigens.

Proteoglycan isolation and analysis

DEFF Research Database (Denmark)

Woods, A; Couchman, J R

2001-01-01

Proteoglycans can be difficult molecules to isolate and analyze due to large mass, charge, and tendency to aggregate or form macromolecular complexes. This unit describes detailed methods for purification of matrix, cell surface, and cytoskeleton-linked proteoglycans. Methods for analysis...

UniProt search blastx result: AK287474 [KOME

Lifescience Database Archive (English)

Full Text Available AK287474 J043022N04 P04917|PGSG_RAT Secretory granule proteoglycan core protein pre...cursor (Chondroitin sulfate proteoglycan core protein) (Proteoglycan 10K core protein) (PG19 core protein) (Cytolytic granule proteoglycan core protein) - Rattus norvegicus (Rat) 0 ...

Transmembrane Signaling Proteoglycans

DEFF Research Database (Denmark)

Couchman, John R

2010-01-01

Virtually all metazoan cells contain at least one and usually several types of transmembrane proteoglycans. These are varied in protein structure and type of polysaccharide, but the total number of vertebrate genes encoding transmembrane proteoglycan core proteins is less than 10. Some core prote...... proteins, including those of the syndecans, always possess covalently coupled glycosaminoglycans; others do not. Syndecan has a long evolutionary history, as it is present in invertebrates, but many other transmembrane proteoglycans are vertebrate inventions. The variety of proteins...... proteins has been obtained in mouse knockout experiments. Here some of the latest developments in the field are examined in hopes of stimulating further interest in this fascinating group of molecules. Expected final online publication date for the Annual Review of Cell and Developmental Biology Volume 26...

Periodate Oxidation for Sulfated Glycosaminoglycans, with Special Reference to the Position of Extra Sulfate Groups in Chondroitin Polysulfates, Chondroitin Sulfate D and Chondroitin Sulfate K

OpenAIRE

Seno, Nobuko; Murakami, Keiko; Shibusawa, Haru

1981-01-01

The optimum conditions for periodate oxidation of sulfated disaccharides were investigated to determine the position of extra sulfate groups on the saturated disulfated disaccharides obtained from chondroitin polysulfates, chondroitin sulfates D and K. Under the conditions: 2mM saturated disulfated disaccharide with 20mM sodium periodate at 37°in the dark, the uronic acid residue in the disulfated disaccharide from chondroitin sulfate D was rapidly and completely destroyed, whereas that in th...

Role of chondroitin sulphate tethered silk scaffold in cartilaginous disc tissue regeneration.

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Bhattacharjee, Maumita; Chawla, Shikha; Chameettachal, Shibu; Murab, Sumit; Bhavesh, Neel Sarovar; Ghosh, Sourabh

2016-04-12

Strategies for tissue engineering focus on scaffolds with tunable structure and morphology as well as optimum surface chemistry to simulate the anatomy and functionality of the target tissue. Silk fibroin has demonstrated its potential in supporting cartilaginous tissue formation both in vitro and in vivo. In this study, we investigate the role of controlled lamellar organization and chemical composition of biofunctionalized silk scaffolds in replicating the structural properties of the annulus region of an intervertebral disc using articular chondrocytes. Covalent attachment of chondroitin sulfate (CS) to silk is characterized. CS-conjugated silk constructs demonstrate enhanced cellular metabolic activity and chondrogenic redifferentiation potential with significantly improved mechanical properties over silk-only constructs. A matrix-assisted laser desorption ionization-time of flight analysis and protein-protein interaction studies help to generate insights into how CS conjugation can facilitate the production of disc associated matrix proteins, compared to a silk-only based construct. An in-depth understanding of the interplay between such extra cellular matrix associated proteins should help in designing more rational scaffolds for cartilaginous disc regeneration needs.

Role of chondroitin sulphate tethered silk scaffold in cartilaginous disc tissue regeneration

International Nuclear Information System (INIS)

Bhattacharjee, Maumita; Chawla, Shikha; Chameettachal, Shibu; Murab, Sumit; Ghosh, Sourabh; Bhavesh, Neel Sarovar

2016-01-01

Strategies for tissue engineering focus on scaffolds with tunable structure and morphology as well as optimum surface chemistry to simulate the anatomy and functionality of the target tissue. Silk fibroin has demonstrated its potential in supporting cartilaginous tissue formation both in vitro and in vivo. In this study, we investigate the role of controlled lamellar organization and chemical composition of biofunctionalized silk scaffolds in replicating the structural properties of the annulus region of an intervertebral disc using articular chondrocytes. Covalent attachment of chondroitin sulfate (CS) to silk is characterized. CS-conjugated silk constructs demonstrate enhanced cellular metabolic activity and chondrogenic redifferentiation potential with significantly improved mechanical properties over silk-only constructs. A matrix-assisted laser desorption ionization-time of flight analysis and protein–protein interaction studies help to generate insights into how CS conjugation can facilitate the production of disc associated matrix proteins, compared to a silk-only based construct. An in-depth understanding of the interplay between such extra cellular matrix associated proteins should help in designing more rational scaffolds for cartilaginous disc regeneration needs. (paper)

Biomimetic Proteoglycan Interactions with Type I Collagen Investigated via 2D and 3D TEM

Science.gov (United States)

Moorehead, Carli

Collagen is one of the leading components in extracellular matrix (ECM), providing durability, structural integrity, and functionality for many tissues. Regulation of collagen fibrillogenesis and degradation is important in the treatment of a number of diseases from orthopedic injuries to genetic deficiencies. Recently, novel, biocompatible, semi-synthetic biomimetic proteoglycans (BPGs) were developed, which consist of an enzymatically resistant synthetic polymer core and natural chondroitin sulfate bristles. It was demonstrated that BPGs affect type I collagen fibrillogenesis in vitro, as reflected by their impact delaying the kinetic formation of gels similar to native PGs. This indicates that the morphology of collagen scaffolds as well as endogenous ECM could also be modulated by these proteoglycan mimics. However, the imaging modality used previously, reflectance confocal microscopy, did not yield the resolution necessary to spatially localize BPGs within the collagen network or investigate the effect of BPGs on the quality of collagen fibrils produced in an in vitro fibrillogenesis model which is important for understanding the method of interaction. Consequently, a histological technique, electron tomography, was adapted and utilized to 3D image the nano-scale structures within this simplified tissue model. BPGs were found to aid in lateral growth and enhance fibril banding periodicity resulting in structures more closely resembling those in tissue, in addition to attaching to the collagen surface despite the lack of a protein core.

Chondroitin / dermatan sulfate modification enzymes in zebrafish development.

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Judith Habicher

Full Text Available Chondroitin/dermatan sulfate (CS/DS proteoglycans consist of unbranched sulfated polysaccharide chains of repeating GalNAc-GlcA/IdoA disaccharide units, attached to serine residues on specific proteins. The CS/DS proteoglycans are abundant in the extracellular matrix where they have essential functions in tissue development and homeostasis. In this report a phylogenetic analysis of vertebrate genes coding for the enzymes that modify CS/DS is presented. We identify single orthologous genes in the zebrafish genome for the sulfotransferases chst7, chst11, chst13, chst14, chst15 and ust and the epimerase dse. In contrast, two copies were found for mammalian sulfotransferases CHST3 and CHST12 and the epimerase DSEL, named chst3a and chst3b, chst12a and chst12b, dsela and dselb, respectively. Expression of CS/DS modification enzymes is spatially and temporally regulated with a large variation between different genes. We found that CS/DS 4-O-sulfotransferases and 6-O-sulfotransferases as well as CS/DS epimerases show a strong and partly overlapping expression, whereas the expression is restricted for enzymes with ability to synthesize di-sulfated disaccharides. A structural analysis further showed that CS/DS sulfation increases during embryonic development mainly due to synthesis of 4-O-sulfated GalNAc while the proportion of 6-O-sulfated GalNAc increases in later developmental stages. Di-sulfated GalNAc synthesized by Chst15 and 2-O-sulfated GlcA/IdoA synthesized by Ust are rare, in accordance with the restricted expression of these enzymes. We also compared CS/DS composition with that of heparan sulfate (HS. Notably, CS/DS biosynthesis in early zebrafish development is more dynamic than HS biosynthesis. Furthermore, HS contains disaccharides with more than one sulfate group, which are virtually absent in CS/DS.

A cell surface chondroitin sulfate proteoglycan, immunologically related to CD44, is involved in type I collagen-mediated melanoma cell motility and invasion

DEFF Research Database (Denmark)

Faassen, A E; Schrager, J A; Klein, D J

1992-01-01

The metastatic spread of tumor cells occurs through a complex series of events, one of which involves the adhesion of tumor cells to extracellular matrix (ECM) components. Multiple interactions between cell surface receptors of an adherent tumor cell and the surrounding ECM contribute to cell...... collagen could also be inhibited by removing cell surface chondroitin sulfate with chondroitinase. In contrast, type I collagen-mediated melanoma cell adhesion and spreading were not affected by either beta-D-xyloside or chondroitinase treatments. These results suggest that mouse melanoma CSPG...... was shown to be mediated, at least in part, by chondroitin sulfate. Additionally we have determined that mouse melanoma CSPG is composed of a 110-kD core protein that is recognized by anti-CD44 antibodies on Western blots. Collectively, our data suggests that interactions between a cell surface CD44-related...

Chondroitin Sulfate Is Indispensable for Pluripotency and Differentiation of Mouse Embryonic Stem Cells

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Izumikawa, Tomomi; Sato, Ban; Kitagawa, Hiroshi

2014-01-01

Chondroitin sulfate (CS) proteoglycans are present on the surfaces of virtually all cells and in the extracellular matrix and are required for cytokinesis at early developmental stages. Studies have shown that heparan sulfate (HS) is essential for maintaining mouse embryonic stem cells (ESCs) that are primed for differentiation, whereas the function of CS has not yet been elucidated. To clarify the role of CS, we generated glucuronyltransferase-I-knockout ESCs lacking CS. We found that CS was required to maintain the pluripotency of ESCs and promoted initial ESC commitment to differentiation compared with HS. In addition, CS-A and CS-E polysaccharides, but not CS-C polysaccharides, bound to E-cadherin and enhanced ESC differentiation. Multiple-lineage differentiation was inhibited in chondroitinase ABC-digested wild-type ESCs. Collectively, these results suggest that CS is a novel determinant in controlling the functional integrity of ESCs via binding to E-cadherin.

Synthesis and Characterization of a Chondroitin Sulfate Based Hybrid Bio/Synthetic Biomimetic Aggrecan Macromolecule

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Sarkar, Sumona

Lower back pain resulting from intervertebral disc degeneration is one of the leading musculoskeletal disorders confronting our health system. In order to mechanically stabilize the disc early in the degenerative cascade and prevent the need for spinal fusion surgeries, we have proposed the development of a hybrid-bio/synthetic biomimetic proteoglycan macromolecule for injection into the disc in the early stages of degeneration. The goal of this thesis was to incorporate natural chondroitin sulfate (CS) chains into bottle brush polymer synthesis strategies for the fabrication of CS-macromolecules which mimic the proteoglycan structure and function while resisting enzymatic degradation. Both the "grafting-to" and "grafting-through" techniques of bottle brush synthesis were explored. CS was immobilized via a terminal primary amine onto a model polymeric backbone (polyacrylic acid) for investigation of the "grafting-to" strategy and an epoxy-amine step-growth polymerization technique was utilized for the "grafting-through" synthesis of CS-macromolecules with polyethylene glycol backbone segments. Incorporation of a synthetic polymeric backbone at the terminal amine of CS was confirmed via biochemical assays, 1H-NMR and FTIR spectroscopy, and CS-macromolecule size was demonstrated to be higher than that of natural CS via gel permeation chromatography, transmission electron microscopy and viscosity measurements. Further analysis of CS-macromolecule functionality indicated maintenance of natural CS properties such as high fixed charge density, high osmotic potential and low cytotoxicity with nucleus pulposus cells. These studies are the first attempt at the incorporation of natural CS into biomimetic bottle brush structures. CS-macromolecules synthesized via the methods developed in these studies may be utilized in the treatment and prevention of debilitating back pain as well as act as mimetics for other proteoglycans implicated in cartilage, heart valve, and nervous

SwissProt search result: AK110012 [KOME

Lifescience Database Archive (English)

Full Text Available ulfate proteoglycan 6) (Chromosome segregation protein SmcD) (Bamacan) (Basement membrane-associated chondroitin proteoglycan) (Mad member-interacting protein 1) SMC3_MOUSE 1e-14 ...

SwissProt search result: AK105135 [KOME

Lifescience Database Archive (English)

Full Text Available ulfate proteoglycan 6) (Chromosome segregation protein SmcD) (Bamacan) (Basement membrane-associated chondroitin proteoglycan) (Mad member-interacting protein 1) SMC3_MOUSE 3e-61 ...

SwissProt search result: AK065733 [KOME

Lifescience Database Archive (English)

Full Text Available lfate proteoglycan 6) (Chromosome segregation protein SmcD) (Bamacan) (Basement membrane-associated chondroitin proteoglycan) (Mad member-interacting protein 1) SMC3_MOUSE 4e-18 ...

SwissProt search result: AK242597 [KOME

Lifescience Database Archive (English)

Full Text Available lfate proteoglycan 6) (Chromosome segregation protein SmcD) (Bamacan) (Basement membrane-associated chondroitin proteoglycan) (Mad member-interacting protein 1) SMC3_MOUSE 5e-19 ...

SwissProt search result: AK103514 [KOME

Lifescience Database Archive (English)

Full Text Available lfate proteoglycan 6) (Chromosome segregation protein SmcD) (Bamacan) (Basement membrane-associated chondroitin proteoglycan) (Mad member-interacting protein 1) SMC3_MOUSE 1e-33 ...

SwissProt search result: AK120333 [KOME

Lifescience Database Archive (English)

Full Text Available lfate proteoglycan 6) (Chromosome segregation protein SmcD) (Bamacan) (Basement membrane-associated chondroitin proteoglycan) (Mad member-interacting protein 1) SMC3_MOUSE 2e-18 ...

SwissProt search result: AK064293 [KOME

Lifescience Database Archive (English)

Full Text Available ulfate proteoglycan 6) (Chromosome segregation protein SmcD) (Bamacan) (Basement membrane-associated chondroitin proteoglycan) (Mad member-interacting protein 1) SMC3_MOUSE 2e-15 ...

Dynamic expression of a native chondroitin sulfate epitope reveals microheterogeneity of extracellular matrix organization in the embryonic chick heart.

Science.gov (United States)

Capehart, A A; Mjaatvedt, C H; Hoffman, S; Krug, E L

1999-02-01

TC2 is a novel monoclonal antibody produced by in vitro immunization of splenocytes with a peanut agglutinin-positive fraction from extracts of prechondrogenic micromass cultures of chick limb mesenchyme. ELISA results demonstrated TC2 reactivity with a native epitope on a glycosaminoglycan (GAG) enriched in chondroitin-4-sulfate and with multiple intact proteoglycans, but not with other GAGs tested. TC2 immunohistochemical reactivity was abolished by pretreatment of sections with chondroitinase AC or preadsorption with chondroitin-4-sulfate GAG. Strong TC2 localization occurred throughout the developing heart at stage 9. As looping ensued, a graded reactivity was observed from lowest in the atrium to highest in the conotruncus that correlated well with versican localization. The superior atrioventricular cushion stained preferentially with TC2 as compared to the inferior cushion at stages 16-18. At these later stages TC2 patterns did not agree completely with anti-versican reactivity. By stage 23 there was a marked reduction in TC2 localization in the heart, however, strong reactivity remained at certain sites, including the conotruncus and in subcompartments of both atrioventricular cushions. A heterogeneous distribution of other native chondroitin sulfate glycosaminoglycan epitopes recognized by monoclonal antibodies d1C4 and CS-56 was observed as well. The distribution of the TC2 epitope usually did not overlap with d1C4 or CS-56 localization at the stages examined. Overall, the spatiotemporal characteristics of TC2 reactivity in the developing chick heart appear to correlate with subdomains of the endocardial cushions as well as with trabecular and atrial septal formation.

Genetic variations in genes involved in heparan sulphate biosynthesis are associated with Plasmodium falciparum parasitaemia: a familial study in Burkina Faso

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Atkinson Alexandre

2012-04-01

Full Text Available Abstract Background There is accumulating evidence that host heparan sulphate proteoglycans play an important role in the life cycle of Plasmodium through their heparan sulphate chains, suggesting that genetic variations in genes involved in heparan sulphate biosynthesis may influence parasitaemia. Interestingly, Hs3st3a1 and Hs3st3b1 encoding enzymes involved in the biosynthesis of heparan sulphate are located within a chromosomal region linked to Plasmodium chabaudi parasitaemia in mice. This suggests that HS3ST3A1 and HS3ST3B1 may influence P. falciparum parasitaemia in humans. Methods Polymorphisms within HS3ST3A1 and HS3ST3B1 were identified in 270 individuals belonging to 44 pedigrees and living in Burkina Faso. Linkage and association between parasitaemia and the polymorphisms were assessed with MERLIN and FBAT. A genetic interaction analysis was also conducted based on the PGMDR approach. Results Linkage between P. falciparum parasitaemia and the chromosomal region containing HS3ST3A1 and HS3ST3B1 was detected on the basis of the 20 SNPs identified. In addition, rs28470223 located within the promoter of HS3ST3A1 was associated with P. falciparum parasitaemia, whereas the PGMDR analysis revealed a genetic interaction between HS3ST3A1 and HS3ST3B1. Seventy-three significant multi-locus models were identified after correcting for multiple tests; 37 significant multi-locus models included rs28470223, whereas 38 multi-locus models contained at least one mis-sense mutation within HS3ST3B1. Conclusion Genetic variants of HS3ST3A1 and HS3ST3B1 are associated with P. falciparum parasitaemia. This suggests that those variants alter both the function of heparan sulphate proteoglycans and P. falciparum parasitaemia.

Development of affinity-based delivery of NGF from a chondroitin sulfate biomaterial.

Science.gov (United States)

Butterfield, Karen Chao; Conovaloff, Aaron W; Panitch, Alyssa

2011-01-01

Chondroitin sulfate is a major component of the extracellular matrix in both the central and peripheral nervous systems. Chondroitin sulfate is upregulated at injury, thus methods to promote neurite extension through chondroitin sulfate-rich matrices and synthetic scaffolds are needed. We describe the use of both chondroitin sulfate and a novel chondroitin sulfate-binding peptide to control the release of nerve growth factor. Interestingly, the novel chondroitin sulfate-binding peptide enhances the controlled release properties of the chondroitin sulfate gels. While introduction of chondroitin sulfate into a scaffold inhibits primary cortical outgrowth, the combination of chondroitin sulfate, chondroitin sulfate-binding peptide and nerve growth factor promotes primary cortical neurite outgrowth in chondroitin sulfate gels.

Heparan sulfate proteoglycans undergo differential expression alterations in right sided colorectal cancer, depending on their metastatic character

International Nuclear Information System (INIS)

Fernández-Vega, Iván; García-Suárez, Olivia; García, Beatriz; Crespo, Ainara; Astudillo, Aurora; Quirós, Luis M.

2015-01-01

Heparan sulfate proteoglycans (HSPGs) are complex molecules involved in the growth, invasion and metastatic properties of cancerous cells. This study analyses the alterations in the expression patterns of these molecules in right sided colorectal cancer (CRC), both metastatic and non-metastatic. Twenty right sided CRCs were studied. A transcriptomic approach was used, employing qPCR to analyze both the expression of the enzymes involved in heparan sulfate (HS) chains biosynthesis, as well as the proteoglycan core proteins. Since some of these proteoglycans can also carry chondroitin sulfate (CS) chains, we include the study of the genes involved in the biosynthesis of these glycosaminoglycans. Immunohistochemical techniques were also used to analyze tissue expression of particular genes showing significant expression differences, of potential interest. Changes in proteoglycan core proteins differ depending on their location; those located intracellularly or in the extracellular matrix show very similar alteration patterns, while those located on the cell surface vary greatly depending on the nature of the tumor: glypicans 1, 3, 6 and betaglycan are affected in the non-metastatic tumors, whereas in the metastatic, only glypican-1 and syndecan-1 are modified, the latter showing opposing alterations in levels of RNA and of protein, suggesting post-transcriptional regulation in these tumors. Furthermore, in non-metastatic tumors, polymerization of glycosaminoglycan chains is modified, particularly affecting the synthesis of the tetrasaccharide linker and the initiation and elongation of CS chains, HS chains being less affected. Regarding the enzymes responsible for the modificaton of the HS chains, alterations were only found in non-metastatic tumors, affecting N-sulfation and the isoforms HS6ST1, HS3ST3B and HS3ST5. In contrast, synthesis of the CS chains suggests changes in epimerization and sulfation of the C4 and C2 in both types of tumor. Right sided CRCs show

An immunohistochemical study of the triangular fibrocartilage complex of the wrist: regional variations in cartilage phenotype

Science.gov (United States)

Milz, S; Sicking, B; Sprecher, C M; Putz, R; Benjamin, M

2007-01-01

The triangular fibrocartilage complex (TFCC) transmits load from the wrist to the ulna and stabilizes the distal radioulnar joint. Damage to it is a major cause of wrist pain. Although its basic structure is well established, little is known of its molecular composition. We have analysed the immunohistochemical labelling pattern of the extracellular matrix of the articular disc and the meniscal homologue of the TFCC in nine elderly individuals (age range 69–96 years), using a panel of monoclonal antibodies directed against collagens, glycosaminoglycans, proteoglycans and cartilage oligomeric matrix protein (COMP). Although many of the molecules (types I, III and VI collagen, chondroitin 4 sulphate, dermatan sulphate and keratan sulphate, the oversulphated epitope of chondroitin 6 sulphate, versican and COMP) were found in all parts of the TFCC, aggrecan, link protein and type II collagen were restricted to the articular disc and to entheses. They were thus not a feature of the meniscal homologue. The shift in tissue phenotype within the TFCC, from a fibrocartilaginous articular disc to a more fibrous meniscal homologue, correlates with biomechanical data suggesting that the radial region is stiff and subject to considerable stress concentration. The presence of aggrecan, link protein and type II collagen in the articular disc could explain why the TFCC is destroyed in rheumatoid arthritis, given that it has been suggested that autoimmunity to these antigens results in the destruction of articular cartilage. The differential distribution of aggrecan within the TFCC is likely to be reflected by regional differences in water content and mobility on the radial and ulnar side. This needs to be taken into account in the design of improved MRI protocols for visualizing this ulnocarpal complex of the wrist. PMID:17532798

Age- and gender-related alteration in plasma advanced oxidation protein products (AOPP) and glycosaminoglycan (GAG) concentrations in physiological ageing.

Science.gov (United States)

Komosinska-Vassev, Katarzyna; Olczyk, Pawel; Winsz-Szczotka, Katarzyna; Kuznik-Trocha, Kornelia; Klimek, Katarzyna; Olczyk, Krystyna

2012-02-13

The authors studied the role of increased oxidative stress in the development of oxidative protein damage and extracellular matrix (ECM) components in ageing. The age- and gender-associated disturbances in connective tissue metabolism were evaluated by the plasma chondroitin sulphated glycosaminoglycans (CS-GAG) and non-sulphated GAG-hyaluronan (HA) measurements. Plasma concentration of advanced oxidation protein products (AOPP) was analysed in order to assess oxidative protein damage and evaluate the possible deleterious role of oxidative phenomenon on tissue proteoglycans' metabolism during the physiological ageing process. Sulphated and non-sulphated GAGs as well as AOPP were quantified in plasma samples from 177 healthy volunteers. A linear age-related decline of plasma CS-GAG level was found in this study (r=-0.46; page (r=0.44; page-dependent relationship has been shown in regard to AOPP. AOPP levels significantly increased with age (r=0.63; pphysiological ageing. A significant correlation was found between the concentrations of AOPP and both CS-GAG (r=-0.31; page changes in the ECM are reflected by CS-GAG and HA plasma levels. Strong correlations between AOPP and ECM components indicate that oxidative stress targets protein and non-protein components of the connective tissue matrix during human ageing.

INTRACELLULAR SYNTHESIS OF CHONDROITIN SULFATE

Science.gov (United States)

Dziewiatkowski, Dominic D.

1962-01-01

In autoradiograms of slices of costal cartilage, incubated for 4 hours in a salt solution containing S35-sulfate and then washed extensively and dehydrated, about 85 per cent of the radioactivity was assignable to the chondrocytes. From alkaline extracts of similarly prepared slices of cartilage, 64 to 83 per cent of the total sulfur-35 in the slices was isolated as chondroitin sulfate by chromatography on an anion-exchange resin. In view of the estimate that only about 15 per cent of the radioactivity was in the matrix, the isolation of 64 to 83 per cent of the total sulfur-35 as chondroitin sulfate is a strong argument that the chondrocytes are the loci in which chondroitin sulfate(s) is synthesized. PMID:13888910

Chondroitin 6-O-sulfotransferases are required for morphogenesis of the notochord in the ascidian embryo.

Science.gov (United States)

Nakamura, Jun; Yoshida, Keita; Sasakura, Yasunori; Fujiwara, Shigeki

2014-12-01

Chondroitin sulfate (CS) is a sulfated polysaccharide chain that binds to various core proteins to form proteoglycans. The amount and position of sulfate groups in CS are variable among different tissues, and are determined by specific sulfotransferases. Although the ascidians are the closest relatives of vertebrates, the functions of their sulfotransferases have not been studied. The genome of the ascidian Ciona intestinalis contains eight genes encoding proteins similar to chondroitin 6-O-sulfotransferases (C6STs), which appear to have independently diverged in the ascidian lineage during evolution. Among them, Ci-C6ST-like1 and Ci-C6ST-like7 were predominantly expressed in the developing notochord. In addition, they were weakly expressed in the neural tube. The disruption of either one of them affected the convergent extension movement of notochordal cells. Presumptive notochord cells coming from both sides of the embryo did not intercalate. The results suggest that both of them are necessary. In some cases, the anterior neural tube failed to close. Forced expression of Ci-C6ST-like1 or Ci-C6ST-like7 in the notochord restored the normal intercalation of notochordal cells, indicating that the effects of morpholino oligos are specific. Ci-C6ST-like1 and Ci-C6ST-like7 are required for the morphogenesis of the notochord in the ascidian embryo. © 2014 Wiley Periodicals, Inc.

A novel chondroitin sulfate hydrogel for nerve repair

Science.gov (United States)

Conovaloff, Aaron William

Brachial plexus injuries affect numerous patients every year, with very debilitating results. The majority of these cases are very severe, and involve damage to the nerve roots. To date, repair strategies for these injuries address only gross tissue damage, but do not supply cells with adequate regeneration signals. As a result, functional recovery is often severely lacking. Therefore, a chondroitin sulfate hydrogel that delivers neurotrophic signals to damaged neurons is proposed as a scaffold to support nerve root regeneration. Capillary electrophoresis studies revealed that chondroitin sulfate can physically bind with a variety of neurotrophic factors, and cultures of chick dorsal root ganglia demonstrated robust neurite outgrowth in chondroitin sulfate hydrogels. Outgrowth in chondroitin sulfate gels was greater than that observed in control gels of hyaluronic acid. Furthermore, the chondroitin sulfate hydrogel's binding activity with nerve growth factor could be enhanced by incorporation of a synthetic bioactive peptide, as revealed by fluorescence recovery after photobleaching. This enhanced binding was observed only in chondroitin sulfate gels, and not in hyaluronic acid control gels. This enhanced binding activity resulted in enhanced dorsal root ganglion neurite outgrowth in chondroitin sulfate gels. Finally, the growth of regenerating dorsal root ganglia in these gels was imaged using label-free coherent anti-Stokes scattering microscopy. This technique generated detailed, high-quality images of live dorsal root ganglion neurites, which were comparable to fixed, F-actin-stained samples. Taken together, these results demonstrate the viability of this chondroitin sulfate hydrogel to serve as an effective implantable scaffold to aid in nerve root regeneration.

Glucosamine exposure reduces proteoglycan synthesis in primary human endothelial cells in vitro

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Trine M. Reine

2016-09-01

Full Text Available Purpose: Glucosamine (GlcN supplements are promoted for medical reasons, for example, for patients with arthritis and other joint-related diseases. Oral intake of GlcN is followed by uptake in the intestine, transport in the circulation and thereafter delivery to chondrocytes. Here, it is postulated to have an effect on synthesis and turnover of extracellular matrix constituents expressed by these cells. Following uptake in the intestine, serum levels are transiently increased, and the endothelium is exposed to increased levels of GlcN. We investigated the possible effects of GlcN on synthesis of proteoglycans (PGs, an important matrix component, in primary human endothelial cells. Methods: Primary human endothelial cells were cultured in vitro in medium with 5 mM glucose and 0–10 mM GlcN. PGs were recovered and analysed by western blotting, or by SDS-PAGE, gel chromatography or ion-exchange chromatography of 35S-PGs after 35S-sulphate labelling of the cells. Results: The synthesis and secretion of 35S-PGs from cultured endothelial cells were reduced in a dose- and time-dependent manner after exposure to GlcN. PGs are substituted with sulphated glycosaminoglycan (GAG chains, vital for PG function. The reduction in 35S-PGs was not related to an effect on GAG chain length, number or sulphation, but rather to the total expression of PGs. Conclusion: Exposure of endothelial cells to GlcN leads to a general decrease in 35S-PG synthesis. These results suggest that exposure to high levels of GlcN can lead to decreased matrix synthesis, contrary to what has been claimed by supporters of such supplements.

Chondroitin sulfate synthase-2 is necessary for chain extension of chondroitin sulfate but not critical for skeletal development.

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Ogawa, Hiroyasu; Hatano, Sonoko; Sugiura, Nobuo; Nagai, Naoko; Sato, Takashi; Shimizu, Katsuji; Kimata, Koji; Narimatsu, Hisashi; Watanabe, Hideto

2012-01-01

Chondroitin sulfate (CS) is a linear polysaccharide consisting of repeating disaccharide units of N-acetyl-D-galactosamine and D-glucuronic acid residues, modified with sulfated residues at various positions. Based on its structural diversity in chain length and sulfation patterns, CS provides specific biological functions in cell adhesion, morphogenesis, neural network formation, and cell division. To date, six glycosyltransferases are known to be involved in the biosynthesis of chondroitin saccharide chains, and a hetero-oligomer complex of chondroitin sulfate synthase-1 (CSS1)/chondroitin synthase-1 and chondroitin sulfate synthase-2 (CSS2)/chondroitin polymerizing factor is known to have the strongest polymerizing activity. Here, we generated and analyzed CSS2(-/-) mice. Although they were viable and fertile, exhibiting no overt morphological abnormalities or osteoarthritis, their cartilage contained CS chains with a shorter length and at a similar number to wild type. Further analysis using CSS2(-/-) chondrocyte culture systems, together with siRNA of CSS1, revealed the presence of two CS chain species in length, suggesting two steps of CS chain polymerization; i.e., elongation from the linkage region up to Mr ∼10,000, and further extension. There, CSS2 mainly participated in the extension, whereas CSS1 participated in both the extension and the initiation. Our study demonstrates the distinct function of CSS1 and CSS2, providing a clue in the elucidation of the mechanism of CS biosynthesis.

THYROID HORMONE TREATED ASTROCYTES INDUCE MATURATION OF CEREBRAL CORTICAL NEURONS THROUGH MODULATION OF PROTEOGLYCAN LEVELS

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Romulo Sperduto Dezonne

2013-08-01

Full Text Available Proper brain neuronal circuitry formation and synapse development is dependent on specific cues, either genetic or epigenetic, provided by the surrounding neural environment. Within these signals, thyroid hormones (T3 and T4 play crucial role in several steps of brain morphogenesis including proliferation of progenitor cells, neuronal differentiation, maturation, migration, and synapse formation. The lack of thyroid hormones during childhood is associated with several impair neuronal connections, cognitive deficits, and mental disorders. Many of the thyroid hormones effects are mediated by astrocytes, although the mechanisms underlying these events are still unknown. In this work, we investigated the effect of 3, 5, 3’-triiodothyronine-treated (T3-treated astrocytes on cerebral cortex neuronal differentiation. Culture of neural progenitors from embryonic cerebral cortex mice onto T3-treated astrocyte monolayers yielded an increment in neuronal population, followed by enhancement of neuronal maturation, arborization and neurite outgrowth. In addition, real time PCR assays revealed an increase in the levels of the heparan sulfate proteoglycans, Glypican 1 (GPC-1 and Syndecans 3 e 4 (SDC-3 e SDC-4, followed by a decrease in the levels of the chondroitin sulfate proteoglycan, Versican. Disruption of glycosaminoglycan chains by chondroitinase AC or heparanase III completely abolished the effects of T3-treated astrocytes on neuronal morphogenesis. Our work provides evidence that astrocytes are key mediators of T3 actions on cerebral cortex neuronal development and identified potential molecules and pathways involved in neurite extension; which might eventually contribute to a better understanding of axonal regeneration, synapse formation and neuronal circuitry recover.

212Pb-Labeled Antibody 225.28 Targeted to Chondroitin Sulfate Proteoglycan 4 for Triple-Negative Breast Cancer Therapy in Mouse Models

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Benjamin B. Kasten

2018-03-01

Full Text Available Triple-negative breast cancer (TNBC is an aggressive subtype of breast cancer with a poor prognosis. There is a clinical need for effective, targeted therapy strategies that destroy both differentiated TNBC cells and TNBC cancer initiating cells (CICs, as the latter are implicated in the metastasis and recurrence of TNBC. Chondroitin sulfate proteoglycan 4 (CSPG4 is overexpressed on differentiated tumor cells and CICs obtained from TNBC patient specimens, suggesting that CSPG4 may be a clinically relevant target for the imaging and therapy of TNBC. The purpose of this study was to determine whether α-particle radioimmunotherapy (RIT targeting TNBC cells using the CSPG4-specific monoclonal antibody (mAb 225.28 as a carrier was effective at eliminating TNBC tumors in preclinical models. To this end, mAb 225.28 labeled with 212Pb (212Pb-225.28 as a source of α-particles for RIT was used for in vitro Scatchard assays and clonogenic survival assays with human TNBC cells (SUM159 and 2LMP grown as adherent cells or non-adherent CIC-enriched mammospheres. Immune-deficient mice bearing orthotopic SUM159 or 2LMP xenografts were injected i.v. with the targeted (225.28 or irrelevant isotype-matched control (F3-C25 mAbs, labeled with 99mTc, 125I, or 212Pb for in vivo imaging, biodistribution, or tumor growth inhibition studies. 212Pb-225.28 bound to adherent SUM159 and 2LMP cells and to CICs from SUM159 and 2LMP mammospheres with a mean affinity of 0.5 nM. Nearly ten times more binding sites per cell were present on SUM159 cells and CICs compared with 2LMP cells. 212Pb-225.28 was six to seven times more effective than 212Pb-F3-C25 at inhibiting SUM159 cell and CIC clonogenic survival (p < 0.05. Radiolabeled mAb 225.28 showed significantly higher uptake than radiolabeled mAb F3-C25 in SUM159 and 2LMP xenografts (p < 0.05, and the uptake of 212Pb-225.28 in TNBC xenografts was correlated with target epitope expression. 212Pb-225.28 caused dose

Development of affinity-based delivery of NGF from a chondroitin sulfate biomaterial

OpenAIRE

Butterfield, Karen Chao; Conovaloff, Aaron W.; Panitch, Alyssa

2011-01-01

Chondroitin sulfate is a major component of the extracellular matrix in both the central and peripheral nervous systems. Chondroitin sulfate is upregulated at injury, thus methods to promote neurite extension through chondroitin sulfate-rich matrices and synthetic scaffolds are needed. We describe the use of both chondroitin sulfate and a novel chondroitin sulfate-binding peptide to control the release of nerve growth factor. Interestingly, the novel chondroitin sulfate-binding peptide enhanc...

Molecular dissection of placental malaria protein VAR2CSA interaction with a chemo-enzymatically synthesized chondroitin sulfate library.

Science.gov (United States)

Sugiura, Nobuo; Clausen, Thomas Mandel; Shioiri, Tatsumasa; Gustavsson, Tobias; Watanabe, Hideto; Salanti, Ali

2016-12-01

Placental malaria, a serious infection caused by the parasite Plasmodium falciparum, is characterized by the selective accumulation of infected erythrocytes (IEs) in the placentas of the pregnant women. Placental adherence is mediated by the malarial VAR2CSA protein, which interacts with chondroitin sulfate (CS) proteoglycans present in the placental tissue. CS is a linear acidic polysaccharide composed of repeating disaccharide units of D-glucuronic acid and N-acetyl-D-galactosamine that are modified by sulfate groups at different positions. Previous reports have shown that placental-adhering IEs were associated with an unusually low sulfated form of chondroitin sulfate A (CSA) and that a partially sulfated dodecasaccharide is the minimal motif for the interaction. However, the fine molecular structure of this CS chain remains unclear. In this study, we have characterized the CS chain that interacts with a recombinant minimal CS-binding region of VAR2CSA (rVAR2) using a CS library of various defined lengths and sulfate compositions. The CS library was chemo-enzymatically synthesized with bacterial chondroitin polymerase and recombinant CS sulfotransferases. We found that C-4 sulfation of the N-acetyl-D-galactosamine residue is critical for supporting rVAR2 binding, whereas no other sulfate modifications showed effects. Interaction of rVAR2 with CS is highly correlated with the degree of C-4 sulfation and CS chain length. We confirmed that the minimum structure binding to rVAR2 is a tri-sulfated CSA dodecasaccharide, and found that a highly sulfated CSA eicosasaccharide is a more potent inhibitor of rVAR2 binding than the dodecasaccharides. These results suggest that CSA derivatives may potentially serve as targets in therapeutic strategies against placental malaria.

Self-assembly of the hydrogel polymer chain consisting of chitosan and chondroitin sulphate in the presence of theophylline;Propriedades de higrogeis constituidos de quitosana e sulfato decondroitina na presenca de teofilina intumescidos em diferentes pHs

Energy Technology Data Exchange (ETDEWEB)

Lopes, Lais C.; Piai, Juliana F.; Fajardo, Andre R.; Rubira, Adley F.; Muniz, Edvani C., E-mail: ecmuniz@uem.b [Universidade Estadual de Maringa (GMPC/UEM), PR (Brazil). Grupo de Materiais Polimericos e Compositos

2009-07-01

In this work, polyelectronic complex (PEC) consisting of two polysaccharides were developed. One is chitosan (QT), cationic polymer, produced by the chitin deacetylation and the other is chondroitin sulphate (CS), anionic polymer, extracted from bovine or porcine aorta. The PECs were prepared in the presence of theophylline (TEO) for evaluating the influence of this drug in the polymer chains reorganization, as well as, studying the mechanical properties and release of SC and TEO in aqueous solutions on different pH conditions. By the obtained results, it was observed that the 84QT/15SC/TEO (% in weight) hydrogel is pH responsive because the CS releasing is more effective at pH 8, while the release of the TEO is higher at pH 2. The hydrogel showed mechanical properties more resistant to pH 2, 8 and 10 and this was attributed to interactions between the polymer chains. Finally, the X-rays profile showed the presence of peaks associated to reorganization of the chains in the hydrogel is at times larger than the hydrogel in the absence of solute. (author)

Sulphate solubility and sulphate diffusion in oxide glasses: implications for the containment of sulphate-bearing nuclear wastes

International Nuclear Information System (INIS)

Lenoir, M.

2009-09-01

The thesis deals with sulphate solubility and sulphate diffusion in oxide glasses, in order to control sulphate incorporation and sulphate volatilization in nuclear waste glasses. It was conducted on simplified compositions, in the SiO 2 -B 2 O 3 -R 2 O (R = Li, Na, K, Cs), SiO 2 -B 2 O 3 -BaO and V 2 O 5 -B 2 O 3 -BaO systems. These compositions allowed us to study the influence of the nature of network-modifying ions (Li + , Na + , K + , Cs + or Ba 2+ ) and also of former elements (Si, B, V), on structure and properties of glasses. Sulphate volatility is studied in sodium borosilicate melts using an innovative technique of sulphate quantitation with Raman spectroscopy. This technique is useful to obtain kinetic curves of sulphate volatilization. The establishment of a model to fit these curves leads to the determination of diffusion coefficients of sulphate. These diffusion coefficients can thus be compared to diffusion coefficients of other species, determined by other techniques and presented in the literature. They are also linked to diffusion coefficients in relation with the viscosity of the melts. Concerning sulphate solubility in glasses, it depends on glass composition and on the nature of sulphate incorporated. Sulphate incorporation in alkali borosilicate glasses leads to the formation of a sulphate layer floating on top of the melt. Sulphate incorporation in barium borosilicate and boro-vanadate glasses leads to the crystallization of sulphate species inside the vitreous matrix. Moreover, sulphate solubility is higher in these glasses than in alkali borosilicates. Finally, exchanges between cations present in glasses and cations present in the sulphate phase are also studied. (author)

Bacterial Reduction Of Barium Sulphate By Sulphate-Reducing Bacteria

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Luptáková Alena

2015-12-01

Full Text Available Acid mine drainage (AMD is a worldwide problem leading to contamination of water sources. AMD are characterized by low pH and high content of heavy metals and sulphates. The barium salts application presents one of the methods for the sulphates removing from AMD. Barium chloride, barium hydroxide and barium sulphide are used for the sulphates precipitation in the form of barium sulphate. Because of high investment costs of barium salts, barium sulphide is recycled from barium sulphate precipitates. It can be recycled by thermic or bacterial reduction of barium sulphate. The aim of our study was to verify experimentally the possibility of the bacterial transformation of BaSO4 to BaS by sulphate-reducing bacteria. Applied BaSO4 came from experiments of sulphates removal from Smolnik AMD using BaCl2.

Removal of sulphates from waste waters by sulphate-reducing bacteria

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Luptáková Alena

2002-03-01

Full Text Available are present in almost all types of water, usually as a simple anion SO42-. The sulphates together with hydrogencarbonates and chlorides are principal anions in natural waters. In typical underground and surface waters, the concentration of sulphates is in the range from ten to hundreds milligrams per litre.Nowadays, the importance of the control of sulphate concentration in waste waters increases. According to the Slovak legislation the limit concentration of sulphates in surface and drinking waters is 250 mg.l-1 . In rivers the contents of sulphates increases mainly by the discharge of waste waters, which are coming mainly from chemical, textile, metallurgical, pharmaceutical, paper and mining industry. The concentration of sulphates in these waters is in the order of grams per litre.Many technologies for the sulphates removal from waste waters exist, including biologico-chemical processes. The principle of one of these methods is the reduction of sulphates by sulphate-reducing bacteria to hydrogen-sulphide.The objective of this work was to study the effect of initial sulphates concentration on the activity of anaerobic sulphate reducers as well as the kinetics of the anaerobic sulphate reduction. The batch reactor was used at temperature of 30°C and pH 7,5. Lactate was used as the carbon source.

[Profile of sulphated glycosaminoglycans content in the murine uterus during the different phases of the estrous cycle].

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Gomes, Regina Célia Teixeira; Simões, Ricardo Santos; Soares, José Maria; Nader, Helena Bonciani; Simões, Manuel de Jesus; Baracat, Edmund C

2007-01-01

Identification and quantitation of sulphated glycosaminoglycans (GAGs) in the uterus of female mice during the estrous cycle. Four groups (n = 10 each) of virgin, 100-day old female mice were assembled according to the estrous cycle phase: proestrus, estrus, metaestrus and diestrus. Samples of the median portion of uterine horns were processed for light microscopy examination (H/E and Alcian blue + PAS). The GAGs were extracted and characterized by agarose gel electrophoresis. Data were analyzed by the unpaired Student's t-test. At light microscopy GAGs appear in all layers of the uterus, especially in the endometrium, between collagen fibers, in the basal membrane and around fibroblasts. Biochemical analyses disclosed presence of dermatan sulphate (DS), chondroitin sulphate (CS and heparan sulphate (HS) during all estral cycle phases. There was no clear electrophoretic separation between DS and CS, thus these two GAGs were considered together (DS+CS) (proestrus = 0.854 +/- 0.192; estrus = 1.073 +/- 0.254; metaestrus = 1.003 +/- 0.255; diestrus = 0.632 +/- 0.443 microg/mg). HS was as follows: proestrus = 0.092 +/- 0.097; estrus = 0.180 +/- 0.141; metaestrus = 0.091 +/- 0.046; diestrus = 0.233 +/- 0.147 microg/mg. The uterine content of DS+CS peaked at estrus (estrogenic action) and that of HS at diestrus (progestagen action). Due to a constant turnover process, there are definite alterations in the uterine profile of GAGs content during the estrous cycle in mice, which may be modulated by female sex hormones.

Chondroitin sulfate effects on neural stem cell differentiation.

Science.gov (United States)

Canning, David R; Brelsford, Natalie R; Lovett, Neil W

2016-01-01

We have investigated the role chondroitin sulfate has on cell interactions during neural plate formation in the early chick embryo. Using tissue culture isolates from the prospective neural plate, we have measured neural gene expression profiles associated with neural stem cell differentiation. Removal of chondroitin sulfate from stage 4 neural plate tissue leads to altered associations of N-cadherin-positive neural progenitors and causes changes in the normal sequence of neural marker gene expression. Absence of chondroitin sulfate in the neural plate leads to reduced Sox2 expression and is accompanied by an increase in the expression of anterior markers of neural regionalization. Results obtained in this study suggest that the presence of chondroitin sulfate in the anterior chick embryo is instrumental in maintaining cells in the neural precursor state.

Aortic smooth muscle cell proteoglycan synthesis in relation to atherosclerosis

International Nuclear Information System (INIS)

Edwards, I.J.

1989-01-01

Proteoglycans (PG) are implicated in atherogenesis by their effects on tissue permeability and cell proliferation and their interaction with plasma low density lipoproteins. Using the pigeon model in which an atherosclerosis-susceptible (WC) and -resistant (SR) breed can be compared, PG synthesis by cultured aortic smooth muscle cells was examined by the use of [ 35 S]-sodium sulfate and [ 3 H]-serine or [ 3 H]-glucosamine as labeling precursors. In both SR and WC cells, the majority of newly synthesized PG were secreted into the media. Chondroitin sulfate (CS) PG and dermatan sulfate (DS) PG were the major PG produced. Total PG production was consistently lower in WC compared to SR cultures due in part to reduce PG synthesis but also to degradation of newly synthesized PG. Since increased DS-PG accompanines atherosclerosis progression, experiments were designed to test the hypothesis that macrophages modulate smooth muscle cell metabolism to cause increase DS-PG production. Cultured WC aortic smooth muscle cells were exposed to the media of cholesteryl ester-loaded pigeon peritoneal macrophages or a macrophage cell line P388D1 and the production of PG examined. Increasing concentration of conditioned media from both types of macrophages caused increased incorporation of 35 S-sulfate into secreted PG, but no change in cell-associated PG. Lipopolysaccharide activation of P388D1 cells enhanced the effect

Chondroitin sulfate-derivatized agarose beads: a new system for studying cation binding to glycosaminoglycans

International Nuclear Information System (INIS)

Hunter, G.K.

1987-01-01

Chondroitin sulfate (CS) has been covalently attached to aminoethyl-agarose beads in a carbodiimide-catalyzed reaction. In this process, an amide bond is formed between carboxylate groups on the glycosaminoglycan (GAG) and the primary amine groups of the beads. Under optimal conditions, up to 160 micrograms of CS is attached per milligram of beads. CS-agarose beads have been used to study Ca binding to GAGs. The beads are mixed with a solution containing CaCl 2 and 45 Ca and allowed to sediment under unit gravity. An aliquot of supernatant is then removed and 45 Ca activity is determined to quantitate remaining (free) Ca. Using this system, it was shown that CS binds approximately 0.7 Ca/disaccharide unit at saturation. Under the conditions used, the apparent association constant (KA) is approximately 14 mM. In principle, this derivatization protocol may be used to attach any proteoglycan or GAG (except keratan sulfate) to an insoluble support. CS-agarose beads provide a rapid, simple, and relatively artifact-free system for studying cation-GAG interactions

Chondroitin Sulfate-E Binds to Both Osteoactivin and Integrin αVβ3 and Inhibits Osteoclast Differentiation.

Science.gov (United States)

Miyazaki, Tatsuya; Miyauchi, Satoshi; Anada, Takahisa; Tawada, Akira; Suzuki, Osamu

2015-10-01

Integrins and their ligands have been suggested to be associated with osteoclast-mediated bone resorption. The present study was designed to investigate whether chondroitin sulfate E (CS-E), which is one of the sulfated glycosaminoglycans (GAGs), is involved in osteoactivin (OA) activity, and osteoclast differentiation. The binding affinity of sulfated GAGs to integrin and its ligand was measured using biotin-labeled CS-E, and the osteoclast differentiation was evaluated by tartrate-resistant acid phosphatase staining and a pit formation assay. CS-E as well as CS-B, synthetic chondroitin polysulfate, and heparin inhibited osteoclast differentiation of bone marrow-derived macrophages. Pre-coating of OA to synthetic calcium phosphate-coated plates enhanced the osteoclastic differentiation of RAW264 cells, and addition of a neutralizing antibody to OA inhibited its differentiation. CS-E bound not only to OA, fibronectin, and vitronectin, but also to its receptor integrin αVβ3, and inhibited the direct binding of OA to integrin αVβ3. Furthermore, CS-E blocked the binding of OA to cells and inhibited OA-induced osteoclastic differentiation. On the other hand, heparinase treatment of RAW264 cells inhibited osteoclastic differentiation. Since binding of OA to the cells was inhibited by the presence of heparan sulfate or heparinase treatment of cells, heparan sulfate proteoglycan (HSPG) was also considered to be an OA receptor. Taken together, the present results suggest that CS-E is capable of inhibiting OA-induced osteoclast differentiation by blocking the interaction of OA to integrin αVβ3 and HSPG. © 2015 Wiley Periodicals, Inc.

Inhibitory effect of chondroitin sulfate oligosaccharides on bovine testicular hyaluronidase.

Science.gov (United States)

Kakizaki, Ikuko; Koizumi, Hideyo; Chen, Fengchao; Endo, Masahiko

2015-05-05

Hyaluronan and chondroitin sulfates are prominent components of the extracellular matrices of animal tissues; however, their functions in relation to their oligosaccharide structures have not yet been fully elucidated. The oligosaccharides of hyaluronan and chondroitin sulfate were prepared and used to investigate their effects on the hydrolysis and transglycosylation reactions of bovine testicular hyaluronidase when hyaluronan was used as a substrate. Hydrolysis and transglycosylation activities were assessed in independent reaction systems by analyzing the products by HPLC. The hydrolysis and transglycosylation reactions of bovine testicular hyaluronidase were dose-dependently inhibited by chondroitin sulfate oligosaccharides, but not by hyaluronan or chondroitin oligosaccharides. A kinetic analysis of the hydrolysis reaction using hyaluronan octasaccharide revealed that the inhibition mode by chondroitin sulfate oligosaccharides was competitive. Copyright © 2015 Elsevier Ltd. All rights reserved.

Immunological methods for the detection and determination of connective tissue proteoglycans

DEFF Research Database (Denmark)

Caterson, B; Baker, J R; Christner, J E

1982-01-01

In this paper we report the use of immunological methods for specifically detecting and determining proteoglycan in cartilage and other connective tissues. Antibodies (polyclonal and monoclonal) have been raised against specific components of cartilage proteoglycan aggregates (i.e., proteoglycan...... surrounding invaginating hair follicles. These immunological procedures are currently being used to complement conventional biochemical analyses of proteoglycans found in different connective tissue matrices....

Characterization of a chondroitin sulfate hydrogel for nerve root regeneration

Science.gov (United States)

Conovaloff, Aaron; Panitch, Alyssa

2011-10-01

Brachial plexus injury is a serious medical problem that affects many patients annually, with most cases involving damage to the nerve roots. Therefore, a chondroitin sulfate hydrogel was designed to both serve as a scaffold for regenerating root neurons and deliver neurotrophic signals. Capillary electrophoresis showed that chondroitin sulfate has a dissociation constant in the micromolar range with several common neurotrophins, and this was determined to be approximately tenfold stronger than with heparin. It was also revealed that nerve growth factor exhibits a slightly stronger affinity for hyaluronic acid than for chondroitin sulfate. However, E8 chick dorsal root ganglia cultured in the presence of nerve growth factor revealed that ganglia cultured in chondroitin sulfate scaffolds showed more robust growth than those cultured in control gels of hyaluronic acid. It is hypothesized that, despite the stronger affinity of nerve growth factor for hyaluronic acid, chondroitin sulfate serves as a better scaffold for neurite outgrowth, possibly due to inhibition of growth by hyaluronic acid chains.

Hexagonal-shaped chondroitin sulfate self-assemblies have exalted anti-HSV-2 activity.

Science.gov (United States)

Galus, Aurélia; Mallet, Jean-Maurice; Lembo, David; Cagno, Valeria; Djabourov, Madeleine; Lortat-Jacob, Hugues; Bouchemal, Kawthar

2016-01-20

The initial step in mucosal infection by the herpes simplex virus type 2 (HSV-2) requires its binding to certain glycosaminoglycans naturally present on host cell membranes. We took advantage of this interaction to design biomimetic supramolecular hexagonal-shaped nanoassemblies composed of chondroitin sulfate having exalted anti-HSV-2 activity in comparison with native chondroitin sulfate. Nanoassemblies were formed by mixing hydrophobically-modified chondroitin sulfate with α-cyclodextrin in water. Optimization of alkyl chain length grafted on chondroitin sulfate and the ratio between hydrophobically-modified chondroitin sulfate and α-cyclodextrin showed that more cohesive and well-structured nanoassemblies were obtained using higher α-cyclodextrin concentration and longer alkyl chain lengths. A structure-activity relationship was found between anti-HSV-2 activity and the amphiphilic nature of hydrophobically-modified chondroitin sulfate. Also, antiviral activity of hexagonal nanoassemblies against HSV-2 was further improved in comparison with hydrophobically-modified chondroitin sulfate. This work suggests a new biomimetic formulation approach that can be extended to other heparan-sulfate-dependent viruses. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effect of depletion of interstitial hyaluronan on hydraulic conductance in rabbit knee synovium

Science.gov (United States)

Coleman, P J; Scott, D; Abiona, A; Ashhurst, D E; Mason, R M; Levick, J R

1998-01-01

The hydraulic resistance of the synovial lining to fluid outflow from a joint cavity () is important for the retention of intra-articular lubricant. The resistance has been attributed in part to extracellular glycosaminoglycans, including hyaluronan and chondroitin sulphates. Increased permeability in joints infused with testicular hyaluronidase, which digests both chondroitin sulphates and hyaluronan, supports this view. In this study the importance of interstitial hyaluronan per se was assessed using leech and Streptomyces hyaluronidases, which degrade only hyaluronan. Ringer solution was infused into the knee joint cavity of anaesthetized rabbits for 30 min, with or without hyaluronidase, after which intra-articular pressure (Pj) was raised and the relation between pressure and outflow determined. Treatment with Streptomyces, leech or testicular hyaluronidases increased the fluid escape rates by similar factors, namely 4- to 6-fold. After Streptomyces hyaluronidase treatment the slope d/dPj, which at low pressures represents synovial hydraulic conductance, increased from a control of 0.90 ± 0.20 μl min−1 cmH2O−1 (mean ± s.e.m., n = 6) to 4.52 ± 0.70 μl min−1 cmH2O−1. The slope d/dPj increased to a similar level after testicular hyaluronidase, namely to 4.14 ± 1.06 μl min−1 cmH2O−1 (control, 0.54 ± 0.24 μl min−1 cmH2O−1). Streptomyces and leech hyaluronidases were as effective as testicular hyaluronidase (no statistically significant differences) despite differences in substrate specificity. It was shown using histochemical and immunohistochemical techniques that hyaluronan was removed from the synovium by leech, Streptomyces and testicular hyaluronidases. The binding of antibodies 2-B-6 and 3-B-3 showed that the core proteins of the chondroitin sulphate proteoglycans remained intact after treatment with hyaluronidases, and the binding of 5-D-4 showed that keratan sulphate was unaffected. An azocasein digestion assay confirmed that the

Biological functions of iduronic acid in chondroitin/dermatan sulfate.

Science.gov (United States)

Thelin, Martin A; Bartolini, Barbara; Axelsson, Jakob; Gustafsson, Renata; Tykesson, Emil; Pera, Edgar; Oldberg, Åke; Maccarana, Marco; Malmstrom, Anders

2013-05-01

The presence of iduronic acid in chondroitin/dermatan sulfate changes the properties of the polysaccharides because it generates a more flexible chain with increased binding potentials. Iduronic acid in chondroitin/dermatan sulfate influences multiple cellular properties, such as migration, proliferation, differentiation, angiogenesis and the regulation of cytokine/growth factor activities. Under pathological conditions such as wound healing, inflammation and cancer, iduronic acid has diverse regulatory functions. Iduronic acid is formed by two epimerases (i.e. dermatan sulfate epimerase 1 and 2) that have different tissue distribution and properties. The role of iduronic acid in chondroitin/dermatan sulfate is highlighted by the vast changes in connective tissue features in patients with a new type of Ehler-Danlos syndrome: adducted thumb-clubfoot syndrome. Future research aims to understand the roles of the two epimerases and their interplay with the sulfotransferases involved in chondroitin sulfate/dermatan sulfate biosynthesis. Furthermore, a better definition of chondroitin/dermatan sulfate functions using different knockout models is needed. In this review, we focus on the two enzymes responsible for iduronic acid formation, as well as the role of iduronic acid in health and disease. © 2013 The Authors Journal compilation © 2013 FEBS.

Ultrastructure Organization of Collagen Fibrils and Proteoglycans of Stingray and Shark Corneal Stroma

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Saud A. Alanazi

2015-01-01

Full Text Available We report here the ultrastructural organization of collagen fibrils (CF and proteoglycans (PGs of the corneal stroma of both the stingray and the shark. Three corneas from three stingrays and three corneas from three sharks were processed for electron microscopy. Tissues were embedded in TAAB 031 resin. The corneal stroma of both the stingray and shark consisted of parallel running lamellae of CFs which were decorated with PGs. In the stingray, the mean area of PGs in the posterior stroma was significantly larger than the PGs of the anterior and middle stroma, whereas, in the shark, the mean area of PGs was similar throughout the stroma. The mean area of PGs of the stingray was significantly larger compared to the PGs, mean area of the shark corneal stroma. The CF diameter of the stingray was significantly smaller compared to the CF diameter in the shark. The ultrastructural features of the corneal stroma of both the stingray and the shark were similar to each other except for the CFs and PGs. The PGs in the stingray and shark might be composed of chondroitin sulfate (CS/dermatan sulfate (DS PGs and these PGs with sutures might contribute to the nonswelling properties of the cornea of the stingray and shark.

Purification and characterization of a 3'-phosphoadenylylsulfate:chondroitin 6-sulfotransferase from arterial tissue.

Science.gov (United States)

Hollmann, J; Niemann, R; Buddecke, E

1986-01-01

A 3'-phosphoadenylylsulfate:chondroitin sulfotransferase (EC 2.8.2.5) was purified to homogeneity (about 760-fold) from the cytosolic fraction of calf arterial tissue by Con A-Sepharose, ion exchange and affinity chromatography. The enzyme has a molecular mass of 38000 Da, optimal activity at pH 6.0 (100%) and 7.25 (75%), requires divalent cations for maximal activity (Mn2+ greater than Mg2+, Ca2+) and exhibits specificity towards desulfated chondroitin sulfate and oligosaccharides derived therefrom. The enzyme transfers sulfate groups from [35S]phosphoadenylylsulfate exclusively to C-6 OH groups of N-acetylgalactosamine units of the acceptor substrates. Maximal sulfate transfer occurs at 2mM chondroitin disaccharide units (100%), the transfer rates decreasing with decreasing chain length in the order deca (55%), octa (17%) and hexasaccharides (4%). Lineweaver-Burk plots revealed equal maximal velocities for chondroitin, deca-, octa- and hexasaccharide, but decreasing Km values. Chondroitin 4-sulfate has 21% of the acceptor potency exhibited by chondroitin, whereas dermatan sulfate, heparan sulfate and hyaluronate and the chondroitin tetrasaccharide showed no acceptor properties. Analysis of the reaction products formed by prolonged enzymatic sulfation of a reduced chondroitin hexasaccharide [GlcA-GalNAc]2-GlcA-GalNAc-ol revealed that the preterminal N-acetylgalactosamine from the non-reducing end and the internal N-acetylgalactosamine but not the N-acetylgalactosaminitol were sulfated and that no hexasaccharide disulfate was formed by the action of chondroitin 6-sulfotransferase. Chondroitin 6-sulfotransferase is considered to possess a binding region capable of accommodating a nonsulfated oligosaccharide sequence of at least six sugars and is believed to act in the course of chondroitin sulfate synthesis in cooperation with, but shortly after, the enzymes involved in the chain elongation reaction.

Effect of retinoic acid on proteoglycan turnover in bovine articular cartilage cultures

International Nuclear Information System (INIS)

Campbell, M.A.; Handley, C.J.

1987-01-01

This paper describes proteoglycan catabolism by adult bovine articular cartilage treated with retinoic acid as a means of stimulating the loss of this macromolecule from the extracellular matrix of cartilage. Addition of retinoic acid (10(-12)-10(-6) M) to adult bovine articular cartilage which had been labeled with [ 35 S]sulfate for 6 h after 5 days in culture, resulted in a dose-dependent increase in the rate of loss of 35 S-labeled proteoglycans from the matrix of the tissue. Concomitant with this loss was a decrease in the proteoglycan content of the tissue. Incubation of cultures treated with 1 microM retinoic acid, at 4 degrees C, or with 0.5 mM cycloheximide, resulted in a significant decrease in the rate of retinoic acid-induced loss of proteoglycans and demonstrated cellular involvement in this process. Analysis of the 35 S-labeled proteoglycans remaining in the matrix showed that the percentage of radioactivity associated with the small proteoglycan species extracted from the matrix of articular cartilage explants labeled with [ 35 S]sulfate after 5 days in culture was 15% and this increased to 22% in tissue maintained in medium alone. In tissue treated with 1 microM retinoic acid for 6 days, the percentage of radioactivity associated with the small proteoglycan was 58%. Approximately 93% of the 35 S-labeled proteoglycans released into the medium of control and retinoic acid-treated cultures was recovered in high density fractions after CsCl gradient centrifugation and eluted on Sepharose CL-2B as a broad peak with a Kav of 0.30-0.37. Less than 17% of these proteoglycans was capable of aggregating with hyaluronate. These results indicate that in both control and retinoic acid-treated cultures the larger proteoglycan species is lost to the medium at a greater rate than the small proteoglycan species. The effect of retinoic acid on proteoglycan turnover was shown to be reversible

Comparison of proteins involved in chondroitin sulfate utilization by three colonic Bacteroides species.

OpenAIRE

Lipeski, L; Guthrie, E P; O'Brien, M; Kotarski, S F; Salyers, A A

1986-01-01

Three species of colonic bacteria can ferment the mucopolysaccharide chondroitin sulfate: Bacteroides ovatus, Bacteroides sp. strain 3452A (an unnamed DNA homology group), and B. thetaiotaomicron. Proteins associated with the utilization of chondroitin sulfate by B. thetaiotaomicron have been characterized previously. In this report we compare chondroitin lyases and chondroitin sulfate-associated outer membrane polypeptides of B. ovatus and Bacteroides sp. strain 3452A with those of B. thetai...

Chlorate: a reversible inhibitor of proteoglycan sulfation

International Nuclear Information System (INIS)

Humphries, D.E.; Silbert, J.E.

1988-01-01

Bovine aorta endothelial cells were cultured in medium containing [ 3 H]glucosamine, [ 35 S]sulfate, and various concentrations of chlorate. Cell growth was not affected by 10 mM chlorate, while 30 mM chlorate had a slight inhibitory effect. Chlorate concentrations greater than 10 mM resulted in significant undersulfation of chondroitin. With 30 mM chlorate, sulfation of chondroitin was reduced to 10% and heparan to 35% of controls, but [ 3 H]glucosamine incorporation on a per cell basis did not appear to be inhibited. Removal of chlorate from the culture medium of cells resulted in the rapid resumption of sulfation

Chondroitin sulfate addition to CD44H negatively regulates hyaluronan binding

International Nuclear Information System (INIS)

Ruffell, Brian; Johnson, Pauline

2005-01-01

CD44 is a widely expressed cell adhesion molecule that binds hyaluronan, an extracellular matrix glycosaminoglycan, in a tightly regulated manner. This regulated interaction has been implicated in inflammation and tumor metastasis. CD44 exists in the standard form, CD44H, or as higher molecular mass isoforms due to alternative splicing. Here, we identify serine 180 in human CD44H as the site of chondroitin sulfate addition and show that lack of chondroitin sulfate addition at this site enhances hyaluronan binding by CD44. A CD44H-immunoglobulin fusion protein expressed in HEK293 cells, and CD44H expressed in murine L fibroblast cells were modified by chondroitin sulfate, as determined by reduced sulfate incorporation after chondroitinase ABC treatment. Mutation of serine 180 or glycine 181 in CD44H reduced chondroitin sulfate addition and increased hyaluronan binding, indicating that serine 180 is the site for chondroitin sulfate addition in CD44H and that this negatively regulates hyaluronan binding

Multinuclear nuclear magnetic resonance spectroscopic study of cartilage proteoglycans

Energy Technology Data Exchange (ETDEWEB)

Lerner, L.

1985-01-01

Hyaline cartilage is a composite material whose major function is to withstand compression while retaining flexibility. Its mechanical properties are affected by tissue hydration and ionic composition. Models of the mechanical behavior of cartilage have incorporated certain assumptions about the interactions of the major components of cartilage: collagen, proteoglycans, water, and cations. To determine the validity of these assumption, the authors have used nuclear magnetic resonance spectroscopy (NMR). Two approaches have been used: (a) natural abundance carbon-13 NMR; and (b) NMR of sodium-23, potassium-39, magnesium-25, and calcium-43. Evidence from studies in intact tissues are reinforced by extensive measurements on solutions of proteoglycans and other relevant macromolecules. Based on the measurements of NMR relaxation rates and lineshapes reported here, it is concluded that neither sodium nor potassium interact strongly with bovine nasal proteoglycan aggregates or their substituent glycosaminoglycan chains in solution. Proteoglycans do bind magnesium and calcium. Therefore there is a qualitative difference between monovalent and divalent cations, which is not taken into account by polyelectrolyte models or models for the ionic dependence of mechanical properties. Cation binding to heparin, which has a higher charge density than cartilage proteoglycans, was also studied. The results presented here establish that heparin binds sodium, magnesium, and calcium.

99mTc-labeled chondroitin sulfate-uptake by chondrocytes and cartilage. Potential agent for osteoarthritis imaging?

International Nuclear Information System (INIS)

Sobal, G.; Sinzinger, H.; Menzel, J.

2002-01-01

Aim: Chondroitin sulfate (CS) is an endogenous component of cartilage proteoglycan which could monitor osteoarthritic cartilage degradation after radiolabeling. This substance is used in the treatment of human osteoarthritis as a slow acting symptomatic drug (CONDROSULF; Sanova Pharma, Vienna; Ibsa, Switzerland). Material and Methods: Radiolabeling of CS was performed using 99m TcO 4 -/stannous chloride in 0.50 M sodium acetate buffer at pH 5.0. The quality control of the tracer was performed using ITLC-SG chromatography and 0.2 M saline in 10% ethanol as solvent to detect colloid content. Aluminium oxide IB-F TLC-sheets and ethanol as solvent were used to estimate free pertechnetate. For uptake studies cultured human chondrocytes and age-matched cartilage were used. Uptake of the tracer in chondrocytes was studied in monolayer and in suspension cultures at 37 0 C. Uptake was monitored for a total of 120-180 minutes, samples being drawn every 10 minutes. Because the commercially available drug Condrosulf contains calcium stearate as additive to improve the resorption of the drug, we investigated also the uptake with and without additive. Results: The tracer was stable over 6h period after labeling (95% of the radiochemical purity). In plasma the stability was lower amounting to 75%. Viability of chondrocytes after incubation with either CS-preparation was found by trypan blue exclusion to be above 95 %. Uptake of the tracer performed in monolayer ± additives was low and amounted to 0.5%±0.05%, n=6. The cells were saturated already after an incubation interval of 10 minutes. In suspension cultures a maximal uptake of 1.0%±0.1%, n=6 and 5.9%±0.65%, n=6 was found, without and with additives, respectively, the saturation was achieved after 100 min. Thus, not only the resorption of the drug is enhanced by Ca-stearate, but also uptake increases in presence of this additive. Using human rib cartilage the uptake of the tracer was much higher amounting to 4.9%±2.3%, n

Holothurian Fucosylated Chondroitin Sulfate

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Vitor H. Pomin

2014-01-01

Full Text Available Fucosylated chondroitin sulfate (FucCS is a structurally distinct glycosaminoglycan found in sea cucumber species. It has the same backbone composition of alternating 4-linked glucuronic acid and 3-linked N-acetyl galactosamine residues within disaccharide repeating units as regularly found in mammalian chondroitin sulfates. However, FucCS has also sulfated fucosyl branching units 3-O-linked to the acid residues. The sulfation patterns of these branches vary accordingly with holothurian species and account for different biological actions and responses. FucCSs may exhibit anticoagulant, antithrombotic, anti-inflammatory, anticancer, antiviral, and pro-angiogenic activities, besides its beneficial effects in hemodialysis, cellular growth modulation, fibrosis and hyperglycemia. Through an historical overview, this document covers most of the science regarding the holothurian FucCS. Both structural and medical properties of this unique GAG, investigated during the last 25 years, are systematically discussed herein.

Effects of glucosamine on proteoglycan loss by tendon, ligament and joint capsule explant cultures.

Science.gov (United States)

Ilic, M Z; Martinac, B; Samiric, T; Handley, C J

2008-12-01

To investigate the effect of glucosamine on the loss of newly synthesized radiolabeled large and small proteoglycans by bovine tendon, ligament and joint capsule. The kinetics of loss of (35)S-labeled large and small proteoglycans from explant cultures of tendon, ligament and joint capsule treated with 10mM glucosamine was investigated over a 10-day culture period. The kinetics of loss of (35)S-labeled small proteoglycans and the formation of free [(35)S]sulfate were determined for the last 10 days of a 15-day culture period. The proteoglycan core proteins were analyzed by gel electrophoresis followed by fluorography. The metabolism of tendon, ligament and joint capsule explants exposed to 10mM glucosamine was evaluated by incorporation of [(3)H]serine and [(35)S]sulfate into protein and glycosaminoglycans, respectively. Glucosamine at 10mM stimulated the loss of small proteoglycans from ligament explant cultures. This was due to the increased loss of both macromolecular and free [(35)S]sulfate to the medium indicating that glucosamine affected the release of small proteoglycans as well as their intracellular degradation. The degradation pattern of small proteoglycans in ligament was not affected by glucosamine. In contrast, glucosamine did not have an effect on the loss of large or small proteoglycans from tendon and joint capsule or large proteoglycans from ligament explant cultures. The metabolism of cells in tendon, ligament and joint capsule was not impaired by the presence of 10mM glucosamine. Glucosamine stimulated the loss of small proteoglycans from ligament but did not have an effect on small proteoglycan catabolism in joint capsule and tendon or large proteoglycan catabolism in ligament, tendon or synovial capsule. The consequences of glucosamine therapy at clinically relevant concentrations on proteoglycan catabolism in joint fibrous connective tissues need to be further assessed in an animal model.

Magnetic resonance imaging reflects cartilage proteoglycan degradation in the rabbit knee

International Nuclear Information System (INIS)

Paul, P.K.; O'Byrne, E.; Blancuzzi, V.; Wilson, D.; Gunson, D.; Douglas, F.L.; Wang Jinzhao; Mezrich, R.S.

1991-01-01

Cartilage degeneration in osteoarthritis is initiated by a loss of proteoglycan. Intra-articular injection of papain causes a reversible loss of proteoglycan in rabbit knees. Rabbits were scanned with magnetic resonance imaging (MRI), using a 1.5T Signa superconducting magnet with 3 inch surface coil. Spin echo sequences were performed in the coronal and sagittal planes at 0, 24, 48, and 72 h after intra-articular injection of papain to abtain T 1 , proton density, and T 2 -weighted images. Cartilage proteoglycan content was measured biochemically and histochemically. Reduced articular cartilage thickness in the MR images of papain-treated knees corresponded to changes in cartilage proteoglycan content. (orig.)

HYDROGELS BASED ON POLYMERS OF DEXTRAN TYRAMINE AND TYRAMINE CONJUGATES OF NATURAL POLYMERS

NARCIS (Netherlands)

Feijen, Jan; Karperien, Marcel; Jin, R.; Moreira Teixeira, Liliana; Dijkstra, Pieter J.

2012-01-01

The invention relates to composition comprising a dextran-tyramine conjugate and a conjugate selected from the group consisting of chondroitin sulphate-tyramine, collagen-tyramine, chitosan-tyramine, chitosan-phloretic acid, gelatine-tyramine, heparan sulphate-tyramine, keratin sulphate-tyramine,

HYDROGELS BASED ON POLYMERS OF DEXTRAN TYRAMINE AND TYRAMINE CONJUGATES OF NATURAL POLYMERS

NARCIS (Netherlands)

Karperien, H.B.J.; Jin, R.; Moreira Teixeira, Liliana; Feijen, Jan; Dijkstra, Pieter J.

2011-01-01

The invention relates to composition comprising a dextran-tyramine conjugate and a conjugate selected from the group consisting of chondroitin sulphate-tyramine, collagen-tyramine, chitosan-tyramine, chitosan-phloretic acid, gelatine-tyramine, heparan sulphate-tyramine, keratin sulphate tyramine,

Reduction of the concentration and total amount of keratan sulphate in synovial fluid from patients with osteoarthritis during treatment with piroxicam.

Science.gov (United States)

Carroll, G J; Bell, M C; Laing, B A; McCappin, S; Blumer, C; Leslie, A

1992-01-01

To study the effects of piroxicam on cartilage metabolism in vivo, a three phase (placebo/piroxicam 20 mg/day by mouth/placebo) double blind controlled trial was conducted in patients with osteoarthritis of the knee joint. Twenty one patients were recruited, 19 of whom (11 women, eight men, median age 70 years) completed the treatment schedule. The knee joint under study was aspirated to dryness at four week intervals. Treatment with piroxicam was accompanied by a decrease in the pain score, an improvement in the functional index, and an increased range of movement. Reductions in the concentration (mean (SEM) 120 (6) to 110 (8) micrograms/ml) and the total amount (1.22 (0.34) to 0.99 (0.37) mg) of keratan sulphate, but not the effusion volume (9.4 (2.5) to 8.3 (2.6) ml) were observed during treatment with piroxicam. These findings are consistent with decreased proteoglycan catabolism during treatment with piroxicam. Neither depressed synthesis nor enhanced clearance of degraded proteoglycan fragments can be excluded, however. PMID:1632658

An introduction to proteoglycans and their localization

DEFF Research Database (Denmark)

Couchman, John R; Pataki, Andreea Csilla

2012-01-01

and in vivo location, and have important roles in invertebrate and vertebrate development, maintenance, and tissue repair. Many biologically potent small proteins can bind glycosaminoglycan chains as a key part of their function in the extracellular matrix, at the cell surface, and also in some intracellular...... locations. Therefore, the participation of proteoglycans in disease is receiving increased attention. In this short review, proteoglycan structure, function, and localizations are summarized, with reference to accompanying reviews in this issue as well as other recent literature. Included are some remarks...

[Analysis of chondroitin sulfate content of Cervi Cornu Pantotrichum with different processing methods and different parts].

Science.gov (United States)

Gong, Rui-Ze; Wang, Yan-Hua; Sun, Yin-Shi

2018-02-01

The differences and the variations of chondroitin sulfate content in different parts of Cervi Cornu Pantotrichum(CCP) with different processing methods were investigated. The chondroitin sulfate from velvet was extracted by dilute alkali-concentrated salt method. Next， the chondroitin sulfate was digested by chondroitinase ABC.The contents of total chondroitin sulfate and chondroitin sulfate A， B and C in the samples were determined by high performance liquid chromatography(HPLC).The content of chondroitin sulfate in wax，powder，gauze，bone slices of CCP with freeze-drying processing is 14.13，11.99，1.74，0.32 g·kg⁻¹， respectively. The content of chondroitin sulfate in wax，powder，gauze，bone slices of CCP with boiling processing is 10.71，8.97，2.21，1.40 g·kg⁻¹， respectively. The content of chondroitin sulfate in wax，powder，gauze，bone slices of CCP without blood is 12.47，9.47，2.64，0.07 g·kg⁻¹， respectively. And the content of chondroitin sulfate in wax，powder，gauze，bone slices of CCP with blood is 8.22，4.39，0.87，0.28 g·kg⁻¹ respectively. The results indicated that the chondroitin sulfate content in different processing methods was significantly different.The content of chondroitin sulfate in CCP with freeze-drying is higher than that in CCP with boiling processing.The content of chondroitin sulfate in CCP without blood is higher than that in CCP with blood. The chondroitin sulfate content in differerent paris of the velvet with the same processing methods was arranged from high to low as： wax slices， powder， gauze slices， bone slices. Copyright© by the Chinese Pharmaceutical Association.

Discrepancies in composition and biological effects of different formulations of chondroitin sulfate.

Science.gov (United States)

Martel-Pelletier, Johanne; Farran, Aina; Montell, Eulàlia; Vergés, Josep; Pelletier, Jean-Pierre

2015-03-06

Osteoarthritis is a common, progressive joint disease, and treatments generally aim for symptomatic improvement. However, SYmptomatic Slow-Acting Drugs in Osteoarthritis (SYSADOAs) not only reduce joint pain, but slow structural disease progression. One such agent is chondroitin sulfate-a complex, heterogeneous polysaccharide. It is extracted from various animal cartilages, thus has a wide range of molecular weights and different amounts and patterns of sulfation. Chondroitin sulfate has an excellent safety profile, and although various meta-analyses have concluded that it has a beneficial effect on symptoms and structure, others have concluded little or no benefit. This may be due, at least partly, to variations in the quality of the chondroitin sulfate used for a particular study. Chondroitin sulfate is available as pharmaceutical- and nutraceutical-grade products, and the latter have great variations in preparation, composition, purity and effects. Moreover, some products contain a negligible amount of chondroitin sulfate and among samples with reasonable amounts, in vitro testing showed widely varying effects. Of importance, although some showed anti-inflammatory effects, others demonstrated weak effects, and some instances were even pro-inflammatory. This could be related to contaminants, which depend on the origin, production and purification process. It is therefore vitally important that only pharmaceutical-grade chondroitin sulfate be used for treating osteoarthritis patients.

Effects of epidermal growth factor on neural crest cells in tissue culture

International Nuclear Information System (INIS)

Erickson, C.A.; Turley, E.A.

1987-01-01

Epidermal growth factor (EGF) stimulates the release of hyaluronic acid (HA) and chondroitin sulfate proteoglycan (CSPG) from quail trunk neural crest cultures in a dose-dependent fashion. It also promotes the expression of cell-associated heparan sulfate proteoglycan (HSPG) as detected by immunofluorescence and immunoprecipitation of the 3 H-labeled proteoglycan. Furthermore, EGF stimulates [ 3 H]thymidine incorporation into total cell DNA. These results raise the possibility that EGF or an analogous growth factor is involved in regulation of neural crest cell morphogenesis

Versican V1 Overexpression Induces a Myofibroblast-Like Phenotype in Cultured Fibroblasts.

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Jon M Carthy

Full Text Available Versican, a chondroitin sulphate proteoglycan, is one of the key components of the provisional extracellular matrix expressed after injury. The current study evaluated the hypothesis that a versican-rich matrix alters the phenotype of cultured fibroblasts.The full-length cDNA for the V1 isoform of human versican was cloned and the recombinant proteoglycan was expressed in murine fibroblasts. Versican expression induced a marked change in fibroblast phenotype. Functionally, the versican-expressing fibroblasts proliferated faster and displayed enhanced cell adhesion, but migrated slower than control cells. These changes in cell function were associated with greater N-cadherin and integrin β1 expression, along with increased FAK phosphorylation. The versican-expressing fibroblasts also displayed expression of smooth muscle α-actin, a marker of myofibroblast differentiation. Consistent with this observation, the versican fibroblasts displayed increased synthetic activity, as measured by collagen III mRNA expression, as well as a greater capacity to contract a collagen lattice. These changes appear to be mediated, at least in part, by an increase in active TGF-β signaling in the versican expressing fibroblasts, and this was measured by phosphorylation and nuclear accumulation of SMAD2.Collectively, these data indicate versican expression induces a myofibroblast-like phenotype in cultured fibroblasts.

Biological functions of iduronic acid in chondroitin/dermatan sulfate

OpenAIRE

Thelin, Martin A; Bartolini, Barbara; Axelsson, Jakob; Gustafsson, Renata; Tykesson, Emil; Pera, Edgar; Oldberg, ?ke; Maccarana, Marco; Malmstrom, Anders

2013-01-01

The presence of iduronic acid in chondroitin/dermatan sulfate changes the properties of the polysaccharides because it generates a more flexible chain with increased binding potentials. Iduronic acid in chondroitin/dermatan sulfate influences multiple cellular properties, such as migration, proliferation, differentiation, angiogenesis and the regulation of cytokine/growth factor activities. Under pathological conditions such as wound healing, inflammation and cancer, iduronic acid has diverse...

Extracellular matrix of cultured glial cells: Selective expression of chondroitin 4-sulfate by type-2 astrocytes and their progenitors

International Nuclear Information System (INIS)

Gallo, V.; Bertolotto, A.

1990-01-01

We have studied the extracellular matrix composition of cultured glial cells by immunocytochemistry with different monoclonal and polyclonal antibodies. Double immunofluorescence experiments and metabolic labeling with [3H]glucosamine performed in different types of cerebellar and cortical cultures showed that bipotential progenitors for type-2 astrocytes and for oligodendrocytes synthesize chondroitin sulfate (CS) and deposit this proteoglycan in their extracellular matrix. The distribution of the various [3H]glucosamine-labeled glycosaminoglycans between the intracellular and the extracellular space was different. CS was present both within the cells and in the culture medium, although in different amounts. Bi-potential progenitors became also O4-positive during their development in vitro. At the stage of O4-positivity they were still stained with antibodies against CS. However, when the progenitor cells were maintained in serum-free medium and differentiated into Gal-C-positive oligodendrocytes, they became CS-negative. In the presence of fetal calf serum in the culture medium, the bipotential progenitors differentiated into GFAP-positive type-2 astrocytes. These cells still expressed CS: their Golgi area and their surface were stained with anti-CS antibodies. Staining with monoclonal antibodies specific for different types of CS (4-sulfate, 6-sulfate, and unsulfated) revealed that both bipotential progenitors and type-2 astrocytes synthesized only chondroitin 4-sulfate. Type-1 astrocytes were negative for both the polyclonal and the monoclonal anti-CS antibodies. Finally, type-2 astrocytes and their progenitors were weakly stained with anti-laminin antibodies and unstained with anti-fibronectin. Type-1 astrocytes were positive for both anti-laminin and anti-fibronectin antibodies and appeared to secrete fibronectin in the extracellular space

The Reclamation of Industrial Wastes Inclusive Sulphates by Sulphate-Reducing Bacteria

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Mária Kušnierová

2004-12-01

Full Text Available The objective of our study was to verify experimentally the possibility of using coal mine drainage and gypsum from the „stabilizate“ (the final product from the combustion desulphurisation as the source of sulphate for the cultivation of SRB with the prospect of: purging of mine waste waters inclusive sulphates, recycling of desulphurisation agent (limestone and production of elemental sulphur from hydrogen sulphide. The results confirmed the theoretical assumptions on the use of gypsum, which forms the substantial component of „stabilizate“, as the source of sulphate for sulphate-reducing bacteria, which produce hydrogen sulphide in the process of bacterial reduction of sulphates. They also showed the possibility of recycling the desulphurisation agent – limestone, as well as the realistic alternative of using „stabilizate“ in the production of elemental sulphur which still represents an important raw material needed in chemical, paper or other industries.

Structural elucidation of fucosylated chondroitin sulfates from sea cucumber using FTICR-MS/MS.

Science.gov (United States)

Agyekum, Isaac; Pepi, Lauren; Yu, Yanlei; Li, Junhui; Yan, Lufeng; Linhardt, Robert J; Chen, Shiguo; Amster, I Jonathan

2018-02-01

Fucosylated chondroitin sulfates are complex polysaccharides extracted from sea cucumber. They have been extensively studied for their anticoagulant properties and have been implicated in other biological activities. While nuclear magnetic resonance spectroscopy has been used to extensively characterize fucosylated chondroitin sulfate oligomers, we herein report the first detailed mass characterization of fucosylated chondroitin sulfate using high-resolution Fourier transform ion cyclotron resonance mass spectrometry. The two species of fucosylated chondroitin sulfates considered for this work include Pearsonothuria graeffei (FCS-Pg) and Isostichopus badionotus (FCS-Ib). Fucosylated chondroitin sulfate oligosaccharides were prepared by N-deacetylation-deaminative cleavage of the two fucosylated chondroitin sulfates and purified by repeated gel filtration. Accurate mass measurements obtained from electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry measurements confirmed the oligomeric nature of these two fucosylated chondroitin sulfate oligosaccharides with each trisaccharide repeating unit averaging four sulfates per trisaccharide. Collision-induced dissociation of efficiently deprotonated molecular ions through Na/H + exchange proved useful in providing structurally relevant glycosidic and cross-ring product ions, capable of assigning the sulfate modifications on the fucosylated chondroitin sulfate oligomers. Careful examination of the tandem mass spectrometry of both species deferring in the positions of sulfate groups on the fucose residue (FCS-Pg-3,4- OS) and (FCS-Ib-2,4- OS) revealed cross-ring products 0,2 A αf and 2,4 X 2αf which were diagnostic for (FCS-Pg-3,4- OS) and 0,2 X 2αf diagnostic for (FCS-Ib-2,4- OS). Mass spectrometry and tandem mass spectrometry data acquired for both species varying in oligomer length (dp3-dp15) are presented.

The identification of proteoglycans and glycosaminoglycans in archaeological human bones and teeth.

Science.gov (United States)

Coulson-Thomas, Yvette M; Coulson-Thomas, Vivien J; Norton, Andrew L; Gesteira, Tarsis F; Cavalheiro, Renan P; Meneghetti, Maria Cecília Z; Martins, João R; Dixon, Ronald A; Nader, Helena B

2015-01-01

Bone tissue is mineralized dense connective tissue consisting mainly of a mineral component (hydroxyapatite) and an organic matrix comprised of collagens, non-collagenous proteins and proteoglycans (PGs). Extracellular matrix proteins and PGs bind tightly to hydroxyapatite which would protect these molecules from the destructive effects of temperature and chemical agents after death. DNA and proteins have been successfully extracted from archaeological skeletons from which valuable information has been obtained; however, to date neither PGs nor glycosaminoglycan (GAG) chains have been studied in archaeological skeletons. PGs and GAGs play a major role in bone morphogenesis, homeostasis and degenerative bone disease. The ability to isolate and characterize PG and GAG content from archaeological skeletons would unveil valuable paleontological information. We therefore optimized methods for the extraction of both PGs and GAGs from archaeological human skeletons. PGs and GAGs were successfully extracted from both archaeological human bones and teeth, and characterized by their electrophoretic mobility in agarose gel, degradation by specific enzymes and HPLC. The GAG populations isolated were chondroitin sulfate (CS) and hyaluronic acid (HA). In addition, a CSPG was detected. The localization of CS, HA, three small leucine rich PGs (biglycan, decorin and fibromodulin) and glypican was analyzed in archaeological human bone slices. Staining patterns were different for juvenile and adult bones, whilst adolescent bones had a similar staining pattern to adult bones. The finding that significant quantities of PGs and GAGs persist in archaeological bones and teeth opens novel venues for the field of Paleontology.

Local changes in proteoglycan synthesis during culture are different for normal and osteoarthritic cartilage

NARCIS (Netherlands)

Lafeber, F. P.; van der Kraan, P. M.; van Roy, H. L.; Vitters, E. L.; Huber-Bruning, O.; van den Berg, W. B.; Bijlsma, J. W.

1992-01-01

Proteoglycan synthesis of mild-to-moderate osteoarthritic human knee cartilage was compared with that of normal cartilage of the same donor. Immediately after cartilage was obtained, the synthesis rate of proteoglycans was higher for osteoarthritic cartilage than for normal cartilage. Proteoglycan

Perlecan domain 1 recombinant proteoglycan augments BMP-2 activity and osteogenesis

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DeCarlo Arthur A

2012-09-01

Full Text Available Abstract Background Many growth factors, such as bone morphogenetic protein (BMP-2, have been shown to interact with polymers of sulfated disacharrides known as heparan sulfate (HS glycosaminoglycans (GAGs, which are found on matrix and cell-surface proteoglycans throughout the body. HS GAGs, and some more highly sulfated forms of chondroitin sulfate (CS, regulate cell function by serving as co-factors, or co-receptors, in GF interactions with their receptors, and HS or CS GAGs have been shown to be necessary for inducing signaling and GF activity, even in the osteogenic lineage. Unlike recombinant proteins, however, HS and CS GAGs are quite heterogenous due, in large part, to post-translational addition, then removal, of sulfate groups to various positions along the GAG polymer. We have, therefore, investigated whether it would be feasible to deliver a DNA pro-drug to generate a soluble HS/CS proteoglycan in situ that would augment the activity of growth-factors, including BMP-2, in vivo. Results Utilizing a purified recombinant human perlecan domain 1 (rhPln.D1 expressed from HEK 293 cells with HS and CS GAGs, tight binding and dose-enhancement of rhBMP-2 activity was demonstrated in vitro. In vitro, the expressed rhPln.D1 was characterized by modification with sulfated HS and CS GAGs. Dose-enhancement of rhBMP-2 by a pln.D1 expression plasmid delivered together as a lyophilized single-phase on a particulate tricalcium phosphate scaffold for 6 or more weeks generated up to 9 fold more bone volume de novo on the maxillary ridge in a rat model than in control sites without the pln.D1 plasmid. Using a significantly lower BMP-2 dose, this combination provided more than 5 times as much maxillary ridge augmentation and greater density than rhBMP-2 delivered on a collagen sponge (InFuse™. Conclusions A recombinant HS/CS PG interacted strongly and functionally with BMP-2 in binding and cell-based assays, and, in vivo, the pln.247 expression plasmid

Sulfation of chondroitin. Specificity, degree of sulfation, and detergent effects with 4-sulfating and 6-sulfating microsomal systems

International Nuclear Information System (INIS)

Sugumaran, G.; Silbert, J.E.

1988-01-01

Microsomal preparations from chondroitin 6-sulfate-producing chick embryo epiphyseal cartilage, and from chondroitin 4-sulfate-producing mouse mastocytoma cells, were incubated with UDP-[14C]glucuronic acid and UDP-N-acetylgalactosamine to form non-sulfated proteo[14C]chondroitin. Aliquots of the incubations were then incubated with 3'-phosphoadenylylphosphosulfate (PAPS) in the presence or absence of various detergents. In the absence of detergents, there was good sulfation of this endogenous proteo[14C]chondroitin by the original microsomes from both sources. Detergents, with the exception of Triton X-100, markedly inhibited sulfation in the mast cell system but not in the chick cartilage system. These results indicate that sulfation and polymerization are closely linked on cell membranes and that in some cases this organization can be disrupted by detergents. When aliquots of the original incubation were heat inactivated, and then reincubated with new microsomes from chick cartilage and/or mouse mastocytoma cells plus PAPS, there was no significant sulfation of this exogenous proteo[14C] chondroitin with either system unless Triton X-100 was added. Sulfation of exogenous chondroitin and chondroitin hexasaccharide was compared with sulfation of endogenous and exogenous proteo[14C]chondroitin. Sulfate incorporation into hexasaccharide and chondroitin decreased as their concentrations (based on uronic acid) approached that of the proteo[14C]chondroitin. At the same time, the degree of sulfation in percent of substituted hexosamine increased. However, the degree of sulfation did not reach that of the endogenous proteo[14C]chondroitin. Hexasaccharide and chondroitin sulfation were stimulated by the presence of Triton X-100. However, in contrast to the exogenous proteo[14C]chondroitin, there was some sulfation of hexasaccharide and chondroitin in the absence of this detergent

Effect of compounds with antibacterial activities in human milk on respiratory syncytial virus and cytomegalovirus in vitro.

Science.gov (United States)

Portelli, J; Gordon, A; May, J T

1998-11-01

The effect of some antibacterial compounds present in human milk were tested for antiviral activity against respiratory syncytial virus, Semliki Forest virus and cytomegalovirus. These included the gangliosides GM1, GM2 and GM3, sialyl-lactose, lactoferrin and chondroitin sulphate A, B and C, which were all tested for their ability to inhibit the viruses in cell culture. Of the compounds tested, only the ganglioside GM2, chondroitin sulphate B and lactoferrin inhibited the absorption and growth of respiratory syncytial virus in cell culture, and none inhibited the growth of Semliki Forest virus, indicating that lipid antiviral activity was not associated with any of the gangliosides. While the concentrations of these two compounds required to inhibit respiratory syncytial virus were in excess of those present in human milk, sialyl-lactose concentrations similar to those present in human milk increased the growth of cytomegalovirus. Lactoferrin was confirmed as inhibiting both respiratory syncytial virus and cytomegalovirus growth in culture even when used at lower concentrations than those present in human milk. The antiviral activities of GM2, chondroitin sulphate B and lactoferrin were tested when added to an infant formula. Lactoferrin continued to have antiviral activity against cytomegalovirus, but a lower activity against respiratory syncytial virus; ganglioside GM2 and chondroitin sulphate B still maintained antiviral activity against respiratory syncytial virus.

Galactosaminoglycan Function and Oligosaccharide Structure Determination

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Daniela G. Seidler

2007-01-01

Full Text Available This review will discuss the importance of sequencing long chondroitin sulfate and dermatan sulfate chains specifically derived from decorin. Decorin is a member of the small leucine-rich repeat proteoglycans and ubiquitously expressed primarily in the skin. Sequence information and diverse function of glycosaminoglycans is further influenced by variable expression through the core protein indicating the importance to analyse glycosaminoglycans from specific proteoglycans.

Metabolism of Cartilage Proteoglycans in Health and Disease

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Demitrios H. Vynios

2014-01-01

Full Text Available Cartilage proteoglycans are extracellular macromolecules with complex structure, composed of a core protein onto which a variable number of glycosaminoglycan chains are attached. Their biosynthesis at the glycosaminoglycan level involves a great number of sugar transferases well-orchestrated in Golgi apparatus. Similarly, their degradation, either extracellular or intracellular in lysosomes, involves a large number of hydrolases. A deficiency or malfunction of any of the enzymes participating in cartilage proteoglycan metabolism may lead to severe disease state. This review summarizes the findings regarding this topic.

Glycoprotein and proteoglycan techniques

International Nuclear Information System (INIS)

Beeley, J.G.

1985-01-01

The aim of this book is to describe techniques which can be used to answer some of the basic questions about glycosylated proteins. Methods are discussed for isolation, compositional analysis, and for determination of the primary structure of carbohydrate units and the nature of protein-carbohydrate linkages of glycoproteins and proteoglycans. High resolution NMR is considered, as well as radioactive labelling techniques. (Auth.)

Heparan sulfate proteoglycans on the cell surface: versatile coordinators of cellular functions

DEFF Research Database (Denmark)

Tumova, S; Woods, A; Couchman, J R

2000-01-01

Heparan sulfate proteoglycans are complex molecules composed of a core protein with covalently attached glycosaminoglycan chains. While the protein part determines localization of the proteoglycan on the cell surfaces or in the extracellular matrix, the glycosaminoglycan component, heparan sulfate......, mediates interactions with a variety of extracellular ligands such as growth factors and adhesion molecules. Through these interactions, heparan sulfate proteoglycans participate in many events during cell adhesion, migration, proliferation and differentiation. We are determining the multitude...... of proteoglycan functions, as their intricate roles in many pathways are revealed. They act as coreceptors for growth factors, participate in signalling during cell adhesion, modulate the activity of a broad range of molecules, and partake in many developmental and pathological processes, including tumorigenesis...

Effects of chondroitin sulfate and glucosamine in adult patients with Kaschin-Beck disease

DEFF Research Database (Denmark)

Zhang, Ya-xu; Dong, Wei; Liu, Hui

2010-01-01

The purpose is to investigate the effects of chondroitin sulfate and glucosamine on adult patients with Kaschin-Beck disease (KBD). A total of 80 patients, aged over 40 years, were randomized into two groups receiving either 1,600 mg oral mixture of chondroitin sulfate and glucosamine or placebo......). But the overall mean change in joint space was significant between the two groups (P chondroitin sulfate and glucosamine might play a protective role in preserving articular cartilage and provide...

Insights into the key roles of proteoglycans in breast cancer biology and translational medicine

DEFF Research Database (Denmark)

Theocharis, Achilleas D.; Skandalis, Spyros S.; Neill, Thomas

2015-01-01

of proteoglycans on tumor and stromal cell membranes affects cancer cell signaling, growth and survival, cell adhesion, migration and angiogenesis. Despite the high complexity and heterogeneity of breast cancer, the rapid evolution in our knowledge that proteoglycans are among the key players in the breast tumor...... in the proteoglycans that will be presented herein provides the potential for multiple layers of regulation of breast tumor behavior. This review summarizes recent developments concerning the biology of selected proteoglycans in breast cancer, and presents potential targeted therapeutic approaches based on their novel...

Combined gelatin-chondroitin sulfate hydrogels for controlled release of cationic antibacterial proteins

NARCIS (Netherlands)

Kuijpers, A. J.; Engbers, G. H. M.; Meyvis, T. K. L.; de Smedt, S. S. C.; Demeester, J.; Krijgsveld, J.; Zaat, S. A. J.; Dankert, J.; Feijen, J.

2000-01-01

Chemically cross-linked gelatin-chondroitin sulfate (ChS) hydrogels were prepared for the controlled release of small cationic proteins. The amount of chondroitin sulfate in the gelatin gels varied between 0 and 20 wt %. The chemical cross-link density, the degree of swelling, and the rheological

Arabidopsis CDS blastp result: AK105135 [KOME

Lifescience Database Archive (English)

Full Text Available AK105135 001-102-A05 At2g27170.1 structural maintenance of chromosomes (SMC) family protein similar to basem...ent membrane-associated chondroitin proteoglycan Bamacan [Rattus norvegicus] GI:178

The identification of proteoglycans and glycosaminoglycans in archaeological human bones and teeth.

Directory of Open Access Journals (Sweden)

Yvette M Coulson-Thomas

Full Text Available Bone tissue is mineralized dense connective tissue consisting mainly of a mineral component (hydroxyapatite and an organic matrix comprised of collagens, non-collagenous proteins and proteoglycans (PGs. Extracellular matrix proteins and PGs bind tightly to hydroxyapatite which would protect these molecules from the destructive effects of temperature and chemical agents after death. DNA and proteins have been successfully extracted from archaeological skeletons from which valuable information has been obtained; however, to date neither PGs nor glycosaminoglycan (GAG chains have been studied in archaeological skeletons. PGs and GAGs play a major role in bone morphogenesis, homeostasis and degenerative bone disease. The ability to isolate and characterize PG and GAG content from archaeological skeletons would unveil valuable paleontological information. We therefore optimized methods for the extraction of both PGs and GAGs from archaeological human skeletons. PGs and GAGs were successfully extracted from both archaeological human bones and teeth, and characterized by their electrophoretic mobility in agarose gel, degradation by specific enzymes and HPLC. The GAG populations isolated were chondroitin sulfate (CS and hyaluronic acid (HA. In addition, a CSPG was detected. The localization of CS, HA, three small leucine rich PGs (biglycan, decorin and fibromodulin and glypican was analyzed in archaeological human bone slices. Staining patterns were different for juvenile and adult bones, whilst adolescent bones had a similar staining pattern to adult bones. The finding that significant quantities of PGs and GAGs persist in archaeological bones and teeth opens novel venues for the field of Paleontology.

Two fucosylated chondroitin sulfates from the sea cucumber Eupentacta fraudatrix.

Science.gov (United States)

Ustyuzhanina, Nadezhda E; Bilan, Maria I; Dmitrenok, Andrey S; Nifantiev, Nikolay E; Usov, Anatolii I

2017-05-15

Two fucosylated chondroitin sulfates EF1 and EF2 were isolated from the sea cucumber Eupentacta fraudatrix. Separation of the polysaccharides was performed using anion-exchange chromatography on DEAE-Sephacel by elution of 0.75M and 1.0M NaCl solutions. The structures of biopolymers were determined by chemical and NMR spectroscopic methods. The backbone of EF1 was found to be composed of chondroitin sulfate A and E units in a ratio of about 1:1. The core of EF2 along with chondroitin sulfate A and E fragments contained unusual disaccharide repeating units →4)-β-d-GlcpA2S3S-(1→3)-β-d-GalpNAc6S-(1→. The main type of branches in both polysaccharides was α-l-Fucp3S4S unit attached to O-3 of GlcA residues. Another type of branches was found to be the disaccharide fragment α-l-Fucp-(1→2)-α-l-Fucp3S4S-(1→ linked to O-3 of GlcA. The presence of structurally different fucosylated chondroitin sulfates in one species of sea cucumber is rather unusual and has not been described previously. Copyright © 2017 Elsevier Ltd. All rights reserved.

Photorefractive keratectomy: measuring the matrix metalloproteinase activity and chondroitin sulfate concentration in tear fluid

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Tetsuya Mutoh

2010-09-01

Full Text Available Tetsuya Mutoh, Masaya Nishio, Yukihiro Matsumoto, Kiyomi Arai, Makoto ChikudaDepartment of Ophthalmology, Dokkyo Medical University Koshigaya Hospital, Saitama, JapanAbstract: We herein report the case of a 20-year-old man who underwent a photorefractive keratectomy (PRK. We measured matrix metalloproteinase-9 (MMP-9 activity and chondroitin 4 sulfate and chondroitin 6 sulfate concentrations in tear fluid. Tear fluid was collected preoperatively via microcapillary tube, and was collected postoperatively on the first and fourth days, and after one week, one month, three months, and six months. Samples were formulated by dilution with 200 µL of saline. MMP-9 activity was analyzed by an enzyme immunocapture activity assay, and the concentrations of chondroitin sulfate were analyzed by enzyme-linked immunosorbent assay. No complications were observed after surgery, except for a minimal subepithelial haze. Although MMP-9 activity changed on the fourth postoperative day, the activity changed only minimally at this time. Chondroitin 4 sulfate concentrations in tear fluid increased dramatically from one week to one month, decreased transiently at three months, and increased by six months. The chondroitin 6 sulfate concentration did not normalize within one week, and decreased from one week to three months compared with the preoperative score, and was close to the preoperative score at six months. We conclude that corneal wound healing was still incomplete six months after PRK, and chondroitin 4 sulfate appears to be critical in this process.Keywords: matrix metalloproteinase, chondroitin sulfate, human tear fluid, photorefractive keratectomy, corneal wound healing

Effects of oversulfated and fucosylated chondroitin sulfates on coagulation. Challenges for the study of anticoagulant polysaccharides.

Science.gov (United States)

Fonseca, Roberto J C; Oliveira, Stephan-Nicollas M C G; Pomin, Vitor H; Mecawi, André S; Araujo, Iracema G; Mourão, Paulo A S

2010-05-01

We report the effects of a chemically oversulfated chondroitin sulfate and a naturally fucosylated chondroitin sulfate on the coagulation system. The former has been recently identified as a contaminant of heparin preparations and the latter has been proposed as an alternative anticoagulant. The mechanism of action of these polymers on coagulation is complex and target different components of the coagulation system. They have serpin-independent anticoagulant activity, which preponderates in plasma. They also have serpin-dependent anticoagulant activity but differ significantly in the target coagulation protease and preferential serpin. Their anticoagulant effects differ even more markedly when tested as inhibitors of coagulation proteases using plasma as a source of serpins. It is possible that the difference is due to the high availability of fucosylated chondroitin sulfate whereas oversulfated chondroitin sulfate has strong unspecific binding to plasma protein and low availability for the binding to serpins. When tested using a venous thrombosis experimental model, oversulfated chondroitin sulfate is less potent as an antithrombotic agent than fucosylated chondroitin sulfate. These highly sulfated chondroitin sulfates activate factor XII in in vitro assays, based on kallikrein release. However, only fucosylated chondroitin sulfate induces hypotension when intravenously injected into rats. In conclusion, the complexity of the regulatory mechanisms involved in the action of highly sulfated polysaccharides in coagulation requires their analysis by a combination of in vitro and in vivo assays. Our results are relevant due to the urgent need for new anticoagulant drugs or alternative sources of heparin.

Variations in the Peritrophic Matrix Composition of Heparan Sulphate from the Tsetse Fly, Glossina morsitans morsitans

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Evelyn Rogerson

2018-03-01

Full Text Available Tsetse flies are the principal insect vectors of African trypanosomes—sleeping sickness in humans and Nagana in cattle. One of the tsetse fly species, Glossina morsitans morsitans, is host to the parasite, Trypanosoma brucei, a major cause of African trypanosomiasis. Precise details of the life cycle have yet to be established, but the parasite life cycle involves crossing the insect peritrophic matrix (PM. The PM consists of the polysaccharide chitin, several hundred proteins, and both glycosamino- and galactosaminoglycan (GAG polysaccharides. Owing to the technical challenges of detecting small amounts of GAG polysaccharides, their conclusive identification and composition have not been possible until now. Following removal of PMs from the insects and the application of heparinases (bacterial lyase enzymes that are specific for heparan sulphate (HS GAG polysaccharides, dot blots with a HS-specific antibody showed heparan sulphate proteoglycans (HSPGs to be present, consistent with Glossina morsitans morsitans genome analysis, as well as the likely expression of the HSPGs syndecan and perlecan. Exhaustive HS digestion with heparinases, fluorescent labeling of the resulting disaccharides with BODIPY fluorophore, and separation by strong anion exchange chromatography then demonstrated the presence of HS for the first time and provided the disaccharide composition. There were no significant differences in the type of disaccharide species present between genders or between ages (24 vs. 48 h post emergence, although the HS from female flies was more heavily sulphated overall. Significant differences, which may relate to differences in infection between genders or ages, were evident, however, in overall levels of 2-O-sulphation between sexes and, for females, between 24 and 48 h post-emergence, implying a change in expression or activity for the 2-O-sulphotransferase enzyme. The presence of significant quantities of disaccharides containing the

Anti-tumor Study of Chondroitin Sulfate-Methotrexate Nanogels

Science.gov (United States)

Wang, Jinyu; Zhao, Weibo; Chen, Haixiao; Qin, An; Zhu, Peizhi

2017-10-01

Self-assembly nanogels (NGs) were formed by bioconjugating methotrexate (MTX) with chondroitin sulfate (CS). MTX-CS NGs can greatly enhance the solubility and improve the delivery efficacy of MTX due to the CD44 binding property of CS. Vivo experiments revealed that MTX-CS NGs showed less toxicity than MTX. MTX-CS NGs can improve the anti-tumor effect while reducing the side effects of MTX. Due to their CD44 binding property, chondroitin sulfate-drug conjugates could be a promising and efficient platform for improving the solubility of sparingly soluble drug molecules as well as targeted delivery to cancer cells and tumor tissues.

Glucosamine and chondroitin use in canines for osteoarthritis: A review

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Angel Bhathal

2017-02-01

Full Text Available Osteoarthritis is a slowly progressive and debilitating disease that affects canines of all breeds. Pain and decreased mobility resulting from osteoarthritis often have a negative impact on the affected canine’s quality of life, level of comfort, daily functioning, activity, behaviour, and client-pet companionship. Despite limited and conflicting evidence, the natural products glucosamine hydrochloride (HCl and chondroitin sulfate are commonly recommended by veterinarians for treating osteoarthritis in dogs. There is a paucity of well-designed clinical veterinary studies investigating the true treatment effect of glucosamine and chondroitin. The purposes of this review article are to provide a brief background on glucosamine and chondroitin use in canine osteoarthritis and to critically review the available literature on the role of these products for improving clinical outcomes. Based on critical review, recommendations for practice are suggested and a future study design is proposed.

Cartilage proteoglycans inhibit fibronectin-mediated adhesion

Science.gov (United States)

Rich, A. M.; Pearlstein, E.; Weissmann, G.; Hoffstein, S. T.

1981-09-01

Normal tissues and organs show, on histological examination, a pattern of cellular and acellular zones that is characteristic and unique for each organ or tissue. This pattern is maintained in health but is sometimes destroyed by disease. For example, in mobile joints, the articular surfaces consist of relatively acellular hyaline cartilage, and the joint space is enclosed by a capsule of loose connective tissue with a lining of fibroblasts and macrophages. In the normal joint these cells are confined to the synovial lining and the articular surface remains acellular. In in vitro culture, macrophages and their precursor monocytes are very adhesive, and fibroblasts can migrate and overgrow surfaces such as collagen or plastic used for tissue culture. The fibroblasts adhere to collagen by means of fibronectin, which they synthesize and secrete1. Because the collagen of cartilage is capable of binding serum fibronectin2 and fibronectin is present in cartilage during its development3, these cells should, in theory, slowly migrate from the synovial lining to the articular surface. It is their absence from the articular cartilage in normal circumstances, and then presence in such pathological states as rheumatoid arthritis, that is striking. We therefore set out to determine whether a component of cartilage could prevent fibroblast adherence in a defined adhesion assay. As normal cartilage is composed of 50% proteoglycans and 50% collagen by dry weight4, we tested the possibility that the proteoglycans in cartilage inhibit fibroblast adhesion to collagen. We present here evidence that fibroblast spreading and adhesion to collagenous substrates is inhibited by cartilage proteoglycans.

Properties of aqueous dispersion of chitosan and chondroitin sulfate complex derived from aquatic organisms

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Novikov V. Yu.

2016-09-01

Full Text Available Investigation of production of chondroitin sulfate, chitosan and polyelectrolyte complexes based on them received from the local marine raw materials is relevant from the point of view of developing a comprehensive waste-free technology for natural raw materials processing. The objects of study are chitosan derived from the shell of the Kamchatka crab Paralithodes camtschaticus and chondroitin sulfate derived from cartilage of salmon Salmon salar. To determine the surface tension of polyelectrolyte complex solutions and dispersions the Wilhelmy method has been used, the effective radius of particle dispersion has been calculated by light scattering, measurements of effective viscosity have been carried out under shear deformation. The conditions of formation, surface and rheological properties of the chitosan and chondroitin sulfate complex extracted from aquatic organisms in the Barents Sea have been studied. Obtaining conditions and molar ratios of these polyelectrolytes in which the aqueous dispersion of the complex remains stable for a long time have been established. It has been found that by addition of chondroitin sulfate solution to chitosan solution in molar ratios of 1 : 3; 1 : 6 the dispersion of the polyelectrolyte complex stable for 2 to 3 days has been formed. The polyelectrolyte complex dispersions behave as non-Newtonian pseudoplastic liquid. When the molar ratio of the mixed solution is 1 : 1 (regardless of the sequence of mixing suspension of the polyelectrolyte complex has been formed, then there is precipitation. Equilibrium surface tension of the aqueous dispersion of the polyelectrolyte complex is higher than that of solutions of chondroitin sulfate and chitosan. The effective radius of particles in the complex dispersion has been determined. The effective radius of the particles in the complex dispersion depends on the molar ratio of chondroitin sulfate : chitosan. A qualitative scheme of formation of polyelectrolyte

Stimulation of proteoglycans by IGF I and II in microvessel and large vessel endothelial cells

International Nuclear Information System (INIS)

Bar, R.S.; Dake, B.L.; Stueck, S.

1987-01-01

Endothelial cells were cultured from bovine capillaries and pulmonary arteries, and the effect of insulinlike growth factor (IGF) I and II (multiplication-stimulating activity) and insulin on the synthesis of proteoglycans was determined. IGF I and II stimulated 35 SO 4 incorporation into proteoglycans in a dose-dependent manner in both microvessel and pulmonary artery endothelial cells with maximum threefold increases. In pulmonary artery cells, the IGFs caused a general stimulation of all classes of glycosaminoglycan-containing proteoglycans. In microvessel endothelial cells, the IGFs appeared to preferentially increase heparan sulfate-containing proteoglycans. Insulin, at concentrations up to 10 -6 M, had no effect on the synthesis of proteoglycans in either microvessel or pulmonary arterial endothelial cells. Thus, the IGFs stimulate the synthesis of proteoglycans in both microvessel and large vessel endothelial cells, a property that is not mimicked by insulin. Because vascular endothelial cells are bathed by IGFs in vivo, such IGF-mediated functions are likely to be significant in both the normal physiology of vascular endothelium and in disease states such as diabetes mellitus

Thermodynamic analysis of dust sulphation reactions

Energy Technology Data Exchange (ETDEWEB)

Yang Yongxiang; Jokilaakso, A.

1997-12-31

Sulphation reactions of metal oxides with SO{sub 2} and O. or SO{sub 3} play significant roles in sulphation roasting of sulphide and oxide minerals as well as in desulphurisation process of combustion gases. In metallurgical waste-heat boilers for sulphide smelting, the sulphation of the oxidic flue dust in the atmosphere containing sulphur oxides is an unavoidable process, and the sulphation reactions have to be guided in a controlled way in the proper parts of the gas handling equipment. In this report, some thermodynamic analyses were conducted for the oxide sulphation reactions in relation to sulphide smelting processes. The phase stability of Me-S-O systems especially for oxides - sulphates equilibrium was studied under different thermodynamic conditions of gas compositions and temperatures. The sulphate stability was analysed for an example of gas compositions in the copper flash smelter of Outokumpu Harjavalta Metals Oy, in relation to temperature. In the report, most of the information was from literature. Moreover, a number of thermodynamic computations were carried out with the HSC program, and the constructed phase stability diagrams were compared with those from the literature whenever possible. The maximum temperatures for stable sulphates under normal operating conditions of the waste-heat boilers in sulphide smelting processes were obtained. This report will serve as the basis for the kinetic studies of the sulphation reactions and the sulphation reaction modelling in pyrometallurgical processes. (orig.) SULA 2 Programme. 36 refs.

Basement membrane heparan sulfate proteoglycan from the L2 rat yolk sac carcinoma

DEFF Research Database (Denmark)

Fenger, M; Wewer, U; Albrechtsen, R

1984-01-01

Heparan sulfate proteoglycan from the L2 rat yolk sac carcinoma has been purified and partially characterized. The proteoglycan has an apparent Mr of 750 000, 35% of which represents the core protein. The core protein seems to be homogeneous, whereas the heparan sulfate chains are heterogeneous...... with an Mr of about 50 000-70 000, with 30% of the glucosamine being N-sulfated. Antibodies raised against the core protein of the heparan sulfate proteoglycan reacted with basement membranes of various rat and human tissue....

Chondroitin Sulfate (CS) Lyases: Structure, Function and Application in Therapeutics.

Science.gov (United States)

Rani, Aruna; Patel, Seema; Goyal, Arun

2018-01-01

Glycosaminoglycans (GAGs) such as chondroitin sulfate (CS) are the chief natural polysaccharides which reside in biological tissues mainly in extracellular matrix. These CS along with adhesion molecules and growth factors are involved in central nervous system (CNS) development, cell progression and pathogenesis. The chondroitin lyases are the enzyme that degrade and alter the CS chains and hence modify various signalling pathways involving CS chains. These CS lyases are substrate specific, can precisely manipulate the CS polysaccharides and have various biotechnological, medical and therapeutic applications. These enzymes can be used to produce the unsaturated oligosaccharides, which have immune-modulatory, anti-inflammatory and neuroprotective properties. This review focuses on the major breakthrough of the chondroitin sulfate degrading enzymes, their structures and functioning mechanism. This also provides comprehensive information regarding production, purification, characterization of CS lyases and their major applications, both established as well as emerging ones such as neural development. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Evaluation of influence of proteoglycans on hydration of articular cartilage with the use of ultrasound

Directory of Open Access Journals (Sweden)

Yi-yi YANG

2015-04-01

Full Text Available Objective　To monitor the changes in hydration behaviour of articular cartilage induced by degradation of proteoglycans, and to explore the effect of proteoglycans on hydration behaviour of articular cartilage by using high-frequency ultrasound. Methods　Twelve porcine patellae with smooth cartilage surface were prepared and equally divided into two groups: normal group without any enzyme treatment, and trypsin group they were treated with 0.25% trypsin for 8h to digest proteoglycan in the cartilage. The hydration behaviour of the cartilage tissue was scanned by high-frequency ultrasound system with a central frequency of 25MHz. Parameters including cartilage hydration strain and cartilage thickness were measured. The histopathological changes in the articular cartilage were observed under a light microscope. Results　It took approximately 20min to reach equilibrium during the hydration process in the normal cartilages, while proteoglycan-degraded cartilage took only about 5min to achieve equilibrium. The equilibrium strain of normal cartilage was 3.5%±0.5%. The degradation of proteoglycans induced a significant decrease in equilibrium strain (1.8%±0.2%, P0.05. Conclusion　Proteoglycans play an important role in hydration behaviour of articular cartilage. The degradation of proteoglycans could induce degeneration of cartilage structure and decrease in hydration behaviour after dehydration. DOI: 10.11855/j.issn.0577-7402.2015.03.03

Intermittent hydrostatic compressive force stimulates exclusively the proteoglycan synthesis of osteoarthritic human cartilage

NARCIS (Netherlands)

Lafeber, F.; Veldhuijzen, J. P.; Vanroy, J. L.; Huber-Bruning, O.; Bijlsma, J. W.

1992-01-01

In paired observations the in vitro proteoglycan turnover was studied of human normal and osteoarthritic cartilage in the absence and presence of intermittent hydrostatic compressive force. Shortly after collection, osteoarthritic cartilage showed a higher proteoglycan synthesis rate than normal

Sulphate removal from sodium sulphate-rich brine and recovery of barium as a barium salt mixture.

Science.gov (United States)

Vadapalli, Viswanath R K; Zvimba, John N; Mulopo, Jean; Motaung, Solly

2013-01-01

Sulphate removal from sodium sulphate-rich brine using barium hydroxide and recovery of the barium salts has been investigated. The sodium sulphate-rich brine treated with different dosages of barium hydroxide to precipitate barium sulphate showed sulphate removal from 13.5 g/L to less than 400 mg/L over 60 min using a barium to sulphate molar ratio of 1.1. The thermal conversion of precipitated barium sulphate to barium sulphide achieved a conversion yield of 85% using coal as both a reducing agent and an energy source. The recovery of a pure mixture of barium salts from barium sulphide, which involved dissolution of barium sulphide and reaction with ammonium hydroxide resulted in recovery of a mixture of barium carbonate (62%) and barium hydroxide (38%), which is a critical input raw material for barium salts based acid mine drainage (AMD) desalination technologies. Under alkaline conditions of this barium salt mixture recovery process, ammonia gas is given off, while hydrogen sulfide is retained in solution as bisulfide species, and this provides basis for ammonium hydroxide separation and recovery for reuse, with hydrogen sulfide also recoverable for further industrial applications such as sulfur production by subsequent stripping.

Inflammatory microRNA-194 and -515 attenuate the biosynthesis of chondroitin sulfate during human intervertebral disc degeneration.

Science.gov (United States)

Hu, Bo; Xu, Chen; Tian, Ye; Shi, Changgui; Zhang, Ying; Deng, Lianfu; Zhou, Hongyu; Cao, Peng; Chen, Huajiang; Yuan, Wen

2017-07-25

Intervertebral disc degeneration (IDD) is characterized by dehydration and loss of extracellular matrixes in the nucleus pulposus region. Chondroitin sulfate has been found to be the water-binding molecule that played a key role in IDD. Although investigators have reported that inflammatory cytokines are involved in the reduction of chondroitin sulfate in IDD, but the underlying mechanism is unrevealed. Since chondroitin sulfate synthesis is controlled by chondroitin sulfate glycosyltransferases CHSY-1/2/3 and CSGALNACT-1/2, their functional role and regulatory mechanism in IDD is not fully studied. Here, we set out to investigate the function and regulatory roles of these factors during IDD development. We found that among these chondroitin sulfate glycosyltransferases, CHSY-1/2/3 are significantly down-regulated in severe IDD samples than mild IDD samples. In vitro experiments revealed that Interleukin-1β and Tumor Necrosis Factor-α stimulation led to significant reduction of CHSY-1/2/3 at protein level than mRNA level in NP cells, indicating a post-transcriptional regulatory mechanisms are involved. By computational prediction and analysis, we found that inflammatory cytokines stimulated microRNA-194 and -515 target CHSY-1/2/3 mRNA and significantly interrupt their translation and downstream chondroitin sulfate deposition. Inhibition of microRNA-194 and -515 however, significantly rescued CHSY-1/2/3 expressions and chondroitin sulfate deposition. These findings together demonstrated a vital role of inflammatory stimulated microRNAs in promoting intervertebral disc degeneration by interrupt chondroitin sulfate synthesis, which may provide new insights into the mechanism and therapeutic approaches in IDD.

Platelet lysate and chondroitin sulfate loaded contact lenses to heal corneal lesions.

Science.gov (United States)

Sandri, Giuseppina; Bonferoni, Maria Cristina; Rossi, Silvia; Delfino, Alessio; Riva, Federica; Icaro Cornaglia, Antonia; Marrubini, Giorgio; Musitelli, Giorgio; Del Fante, Claudia; Perotti, Cesare; Caramella, Carla; Ferrari, Franca

2016-07-25

Hemoderivative tear substitutes contain various ephiteliotrophic factors, such as growth factors (GF), involved in ocular surface homeostasis without immunogenic properties. The aim of the present work was the loading of platelet lysate into contact lenses to improve the precorneal permanence of platelet lysate growth factors on the ocular surface to enhance the treatment of corneal lesions. To this purpose, chondroitin sulfate, a sulfated glycosaminoglycan, which is normally present in the extracellular matrix, was associated with platelet lysate. In fact, chondroitin sulfate is capable of electrostatic interaction with positively charged growth factors, in particular, with bFGF, IGF, VEGF, PDGF and TGF-β, resulting in their stabilization and reduced degradation in solution. In the present work, various types of commercially available contact lenses have been loaded with chondroitin sulfate or chondroitin sulfate in association with platelet lysate to achieve a release of growth factors directly onto the corneal surface lesions. One type of contact lenses (PureVision(®)) showed in vitro good proliferation properties towards corneal cells and were able to enhance cut closure in cornea constructs. Copyright © 2016 Elsevier B.V. All rights reserved.

The combined therapy with chondroitin sulfate plus glucosamine sulfate or chondroitin sulfate plus glucosamine hydrochloride does not improve joint damage in an experimental model of knee osteoarthritis in rabbits.

Science.gov (United States)

Roman-Blas, Jorge A; Mediero, Aránzazu; Tardío, Lidia; Portal-Nuñez, Sergio; Gratal, Paula; Herrero-Beaumont, Gabriel; Largo, Raquel

2017-01-05

Osteoarthritis is the most common chronic joint disorder especially during aging. Although with controversies, glucosamine, both in its forms of sulfate and hydrochloride, and chondroitin sulfate are commonly employed to treat osteoarthritis. Due to the modest improve in the symptoms observed in patients treated with these drugs alone, a formulation combining both agents has been considered. The discrepant results achieved for pain control or structural improvement in osteoarthritis patients has been attributed to the quality of chemical formulations or different bias in clinical studies. The current study has been designed to test the effects of two different combined formulations with adequate pharmaceutical grade of these drugs in osteoarthritic joints, and to explore the underlying mechanisms modulated by both formulations in different osteoarthritis target tissues. Knee osteoarthritis was surgically induced in experimental rabbits. Some animals received the combined therapy (CT)1, (chondroitin sulfate 1200mg/day + glucosamine sulfate 1500mg/day), or the CT2 ((chondroitin sulfate 1200mg/day + glucosamine hydrochloride 1500mg/day). Neither CT1 nor CT2 significantly modified the cartilage damage or the synovial inflammation observed in osteoarthritic animals. Treatments were also unable to modify the presence of pro-inflammatory mediators, and the synthesis of metalloproteinases in the cartilage or in the synovium of osteoarthritic animals. Combined therapies did not modify the decrease in the subchondral bone mineral density observed in osteoarthritic rabbits. Therapies of chondroitin sulfate plus glucosamine sulfate or chondroitin sulfate plus glucosamine hydrochloride failed to improve structural damage or to ameliorate the inflammatory profile of joint tissues during experimental osteoarthritis. Published by Elsevier B.V.

Collageneous matrix coatings on titanium implants modified with decorin and chondroitin sulfate: characterization and influence on osteoblastic cells.

Science.gov (United States)

Bierbaum, Susanne; Douglas, Timothy; Hanke, Thomas; Scharnweber, Dieter; Tippelt, Sonja; Monsees, Thomas K; Funk, Richard H W; Worch, Hartmut

2006-06-01

Studies in developmental and cell biology have established the fact that responses of cells are influenced to a large degree by morphology and composition of the extracellular matrix. Goal of this work is to use this basic principle to improve the biological acceptance of implants by modifying the surfaces with components of the extracellular matrix (ECM), utilizing the natural self-assembly potential of collagen in combination with further ECM components in close analogy to the situation in vivo. Aiming at load-bearing applications in bone contact, collagen type I in combination with the proteoglycan decorin and the glycosaminoglycan chondroitin sulfate (CS) was used; fibrillogenesis, fibril morphology, and adsorption of differently composed fibrils onto titanium were assessed. Both decorin and CS could be integrated into the fibrils during fibrillogenesis, the amount bound respectively desorbed depending on the ionic strength of fibrillogenesis buffer. Including decorin always resulted in a significant decrease of fibril diameter, CS in only a slight decrease or even increase, depending on the collagen preparation used. No significant changes in adsorption to titanium could be detected. Osteoblastic cells showed different reactions for cytoskeletal arrangement and osteopontin expression depending on the composition of the ECM, with CS enhancing the osteoblast phenotype.

Sulphate reduction in the Aespoe HRL tunnel

International Nuclear Information System (INIS)

Gustafson, G.; Pedersen, K.; Tullborg, E.L.; Wallin, B.; Wikberg, P.

1995-12-01

Evidence and indications of sulphate reduction based on geological, hydrogeological, groundwater, isotope and microbial data gathered in and around the Aespoe Hard Rock Laboratory tunnel have been evaluated. This integrated investigation showed that sulphate reduction had taken place in the past but is most likely also an ongoing process. Anaerobic sulphate-reducing bacteria can live in marine sediments, in the tunnel sections under the sea and in deep groundwaters, since there is no access to oxygen. The sulphate-reducing bacteria seem to thrive when the Cl - concentration of the groundwater is 4000-6000 mg/l. Sulphate reduction is an in situ process but the resulting hydrogen-sulphide rich water can be transported to other locations. A more vigorous sulphate reduction takes place when the organic content in the groundwater is high (>10 mg/l DOC) which is the case in the sediments and in the groundwaters under the sea. Some bacteria use hydrogen as an electron donor instead of organic carbon and can therefore live in deep environments where access to organic material is limited. The sulphate-reducing bacteria seem to adapt to changing flow situations caused by the tunnel construction relatively fast. Sulphate reduction seems to have occurred and will probably occur where conditions are favourable for the sulphate-reducing bacteria such as anaerobic brackish groundwater with dissolved sulphate and organic carbon or hydrogen. 59 refs, 37 figs, 6 tabs

Sequence determination of synthesized chondroitin sulfate dodecasaccharides.

Science.gov (United States)

Shioiri, Tatsumasa; Tsuchimoto, Jun; Watanabe, Hideto; Sugiura, Nobuo

2016-06-01

Chondroitin sulfate (CS) is a linear acidic polysaccharide composed of repeating disaccharide units of glucuronic acid and N-acetyl-d-galactosamine. The polysaccharide is modified with sulfate groups at different positions by a variety of sulfotransferases. CS chains exhibit various biological and pathological functions by interacting with cytokines and growth factors and regulating their signal transduction. The fine structure of the CS chain defines its specific biological roles. However, structural analysis of CS has been restricted to disaccharide analysis, hampering the understanding of the structure-function relationship of CS chains. Here, we chemo-enzymatically synthesized CS dodecasaccharides having various sulfate modifications using a bioreactor system of bacterial chondroitin polymerase mutants and various CS sulfotransferases. We developed a sequencing method for CS chains using the CS dodecasaccharides. The method consists of (i) labeling a reducing end with 2-aminopyridine (PA), (ii) partial digestion of CS with testicular hyaluronidase, followed by separation of PA-conjugated oligosaccharides with different chain lengths, (iii) limited digestion of these oligosaccharides with chondroitin lyase AC II into disaccharides, followed by labeling with 2-aminobenzamide, (iv) CS disaccharide analysis using a dual-fluorescence HPLC system (reversed-phase ion-pair and ion-exchange chromatography), and (v) estimation of the composition by calculating individual disaccharide ratios. This CS chain sequencing allows characterization of CS-modifying enzymes and provides a useful tool toward understanding the structure-function relationship of CS chains. © The Author 2016. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Macromolecular basis for homocystein-induced changes in proteoglycan structure in growth and arteriosclerosis.

Science.gov (United States)

McCully, K S

1972-01-01

Cell culture monolayers deficient in cystathionine synthetase bound more inorganic sulfate than normal cell monolayers during growth to confluence; this was correlated with the production of granular proteoglycan by the abnormal cells and fibrillar proteoglycan by normal cells. Homocysteine was demonstrated to be an active precursor of esterified sulfate, confirming our previous finding of this sulfation pathway in liver. The cell cultures deficient in cystathionine synthetase were found to assume an abnormal cellular distribution on the surface of the culture dish, resembling the distribution assumed by neoplastic cells with loss of contact inhibition; the degree of abnormality of the cellular distribution was correlated with the amount of granular proteoglycan produced by the cells and the amount of inorganic sulfate binding by the cell monolayers. Pyridoxine was found to increase the growth rate of cell cultures from a patient with pyridoxineresponsive homocystinuria and to increase the production of fibrillar proteoglycan by the cells; no effect of pyridoxine was observed in the cell cultures from a patient who failed to respond to pyridoxine therapy. The findings suggest that the change in macromolecular conformation of cellular proteoglycans from fibrillar to granular is due to increased sulfation of the carbohydrate envelope of the molecule. The significance of the findings is related to the pathogenesis of homocystinuria, the phenomenon of contact inhibition, the action of growth hormone and initiation of arteriosclerotic plaques.

Copper sulphate poisoning in horses

Energy Technology Data Exchange (ETDEWEB)

Bauer, M

1975-01-01

In the archives of the Clinic for Internal Diseases of Domestic Animals at the Veterinary Faculty of Zagreb University some thirty cases of horse disease diagnosed as copper sulphate poisoning were noted. The data correspond in many respects to the clinical findings of copper sulphate poisoning in other domestic animals. A series of experimental horse poisonings were undertaken in order to determine the toxicity of copper sulphate. The research results are as follows: Horses are sensitive to copper sulphate. Even a single application of 0.125 g/kg body weight in 1% concentration by means of incubation into the stomach causes stomach and gut disturbances and other poisoning symptoms. Poisoning occurs in two types: acute and chronic. The former appears after one to three applications of copper sulphate solution and is characterized by gastroenteritis, haemolysis, jaundice and haemoglobinuria with signs of consecutive damage of kidney, liver and other organs. The disease, from the first application to death lasts for two weeks. Chronic poisoning is caused by ingestion of dry copper sulphate in food (1% solution dried on hay or clover) for two or more months. There are chronic disturbances of stomach and gut and loss of weight, and consecutive (three to four) haemolytic crises similar to those of acute poisoning. From the beginning of poisoning to death six or more months can elapse.

Rabbit chondrocytes maintained in serum-free medium. I. Synthesis and secretion of hydrodynamically-small proteoglycans

International Nuclear Information System (INIS)

Malemud, C.J.; Papay, R.S.

1986-01-01

The biosynthesis of sulfated proteoglycan in vitro by rabbit articular chondrocytes in first passage monolayer culture maintained in fetal bovine serum (FBS) or in serum-free conditions was compared. Neosynthesized proteoglycan in the culture medium in the most dense fraction of an associative CsCl density gradient (fraction dAl) declined with increasing time under serum-free conditions, but not when cells were maintained in the presence of serum. After one day, the major peak of incorporated 35 SO 4 in medium fraction dAl eluted as a retarded peak on Sepharose CL-2B, whether cells were maintained under serum-free or serum-containing conditions. The hydrodynamic size of proteoglycan monomer fraction dAlDl obtained after one day of exposure to serum-free culture media was smaller than dAlDl from serum-containing cultures. The hydrodynamic size of dAlDl obtained from serum-free culture media became even progressively smaller after 2 and 3 days' exposure to these conditions. Hydrodynamically small sulfated proteoglycans were identified in the cell-associated dAlDl fraction as early as one day after switching chondrocytes from serum-containing to serum-free medium. Proteoglycan monomer from serum-containing medium reaggregated more efficiently under both conditions. No change in the size of glycosaminoglycan chains was seen in the smaller proteoglycan subpopulations, nor was there any indication of marked changes in the glycosaminoglycan types

In vivo turnover of the basement membrane and other heparan sulfate proteoglycans of rat glomerulus

DEFF Research Database (Denmark)

Beavan, L A; Davies, M; Couchman, J R

1989-01-01

The metabolic turnover of rat glomerular proteoglycans in vivo was investigated. Newly synthesized proteoglycans were labeled during a 7-h period after injecting sodium [35S]sulfate intraperitoneally. At the end of the labeling period a chase dose of sodium sulfate was given. Subsequently......-propanesulfonate-4 M guanidine hydrochloride, a procedure which solubilized greater than 95% of the 35S-labeled macromolecules. Of these 11-13% was immunoprecipitated by an antiserum against heparan sulfate proteoglycan which, in immunolocalization experiments, showed specificity for staining the basement membrane...

Syndecan proteoglycans and cell adhesion

DEFF Research Database (Denmark)

Woods, A; Oh, E S; Couchman, J R

1998-01-01

It is now becoming clear that a family of transmembrane proteoglycans, the syndecans, have important roles in cell adhesion. They participate through binding of matrix ligand to their glycosaminoglycan chains, clustering, and the induction of signaling cascades to modify the internal microfilament...... organization. Syndecans can modulate the type of adhesive responses induced by other matrix ligand-receptor interactions, such as those involving the integrins, and so contribute to the control of cell morphology, adhesion and migration....

Dermatan Sulfate Epimerase 1-Deficient Mice Have Reduced Content and Changed Distribution of Iduronic Acids in Dermatan Sulfate and an Altered Collagen Structure in Skin

DEFF Research Database (Denmark)

Maccarana, M.; Kalamajski, S.; Kongsgaard, M.

2009-01-01

Dermatan sulfate epimerase 1 (DS-epi1) and DS-epi2 convert glucuronic acid to iduronic acid in chondroitin/dermatan sulfate biosynthesis. Here we report on the generation of DS-epi1-null mice and the resulting alterations in the chondroitin/dermatan polysaccharide chains. The numbers of long blocks...... of adjacent iduronic acids are greatly decreased in skin decorin and biglycan chondroitin/dermatan sulfate, along with a parallel decrease in iduronic-2-O-sulfated-galactosamine-4-O-sulfated structures. Both iduronic acid blocks and iduronic acids surrounded by glucuronic acids are also decreased in versican......-derived chains. DS-epi1-deficient mice are smaller than their wild-type littermates but otherwise have no gross macroscopic alterations. The lack of DS-epi1 affects the chondroitin/dermatan sulfate in many proteoglycans, and the consequences for skin collagen structure were initially analyzed. We found...

Radioprotective effects of dextran sulphate in mice

International Nuclear Information System (INIS)

Vacek, A.; Bartonickova, A.; Rotkovska, D.; Palyga, G.F.; Zhukova, N.A.

1981-01-01

Influence of a single i.p. injection of dextran sulphate on radiosensitivity of mice was investigated. The administration of dextran sulphate 24, 48 and 72 hours prior to irradiation increased formation of endogenous colonies of the hemopoietic tissue on the surface of the spleen. DRF calculated from an equieffective exposure for 5 colonies was 1.96 when dextran sulphate was administered 24 hours before irradiation, and 2.25 when dextran sulphate was administered 72 hours before irradiation. The radioprotective effects of dextran sulphate were manifested also in the survival of animals exposed to lethal doses of short-termed as well as long-termed gamma radiation. (orig.) [de

A multi-analytical approach to better assess the keratan sulfate contamination in animal origin chondroitin sulfate

Energy Technology Data Exchange (ETDEWEB)

Restaino, Odile Francesca, E-mail: odilefrancesca.restaino@unina2.it [Department of Experimental Medicine, Section of Biotechnology and Molecular Biology, University of Campania-L.Vanvitelli, ex Second University of Naples, Via De Crecchio 7, 80138, Naples (Italy); Finamore, Rosario, E-mail: rosario.finamore@unina2.it [Department of Experimental Medicine, Section of Biotechnology and Molecular Biology, University of Campania-L.Vanvitelli, ex Second University of Naples, Via De Crecchio 7, 80138, Naples (Italy); Diana, Paola, E-mail: paola.diana@unina2.it [Department of Experimental Medicine, Section of Biotechnology and Molecular Biology, University of Campania-L.Vanvitelli, ex Second University of Naples, Via De Crecchio 7, 80138, Naples (Italy); Marseglia, Mariacarmela, E-mail: marimars84@hotmail.it [Department of Experimental Medicine, Section of Biotechnology and Molecular Biology, University of Campania-L.Vanvitelli, ex Second University of Naples, Via De Crecchio 7, 80138, Naples (Italy); Vitiello, Mario, E-mail: mariovitiello.ita@gmail.com [Department of Experimental Medicine, Section of Biotechnology and Molecular Biology, University of Campania-L.Vanvitelli, ex Second University of Naples, Via De Crecchio 7, 80138, Naples (Italy); Casillo, Angela, E-mail: angela.casillo@unina.it [Department of Chemical Sciences, University of Naples Federico II, Complesso Universitario Monte S. Angelo, Via Cintia 4, 80126, Naples (Italy); Bedini, Emiliano, E-mail: emiliano.bedini@unina.it [Department of Chemical Sciences, University of Naples Federico II, Complesso Universitario Monte S. Angelo, Via Cintia 4, 80126, Naples (Italy); Parrilli, Michelangelo, E-mail: michelangelo.parrilli@unina.it [Department of Biology, University of Naples Federico II, Complesso Universitario Monte S. Angelo, Via Cintia 4, 80126, Naples (Italy); and others

2017-03-15

Chondroitin sulfate is a glycosaminoglycan widely used as active principle of anti-osteoarthritis drugs and nutraceuticals, manufactured by extraction from animal cartilaginous tissues. During the manufacturing procedures, another glycosaminoglycan, the keratan sulfate, might be contemporarily withdrawn, thus eventually constituting a contaminant difficult to be determined because of its structural similarity. Considering the strict regulatory rules on the pureness of pharmaceutical grade chondrotin sulfate there is an urgent need and interest to determine the residual keratan sulfate with specific, sensitive and reliable methods. To pursue this aim, in this paper, for the first time, we set up a multi-analytical and preparative approach based on: i) a newly developed method by high performance anion-exchange chromatography with pulsed amperometric detection, ii) gas chromatography-mass spectrometry analyses, iii) size exclusion chromatography analyses coupled with triple detector array module and on iv) strong anion exchange chromatography separation. Varied KS percentages, in the range from 0.1 to 19.0% (w/w), were determined in seven pharmacopeia and commercial standards and nine commercial samples of different animal origin and manufacturers. Strong anion exchange chromatography profiles of the samples showed three or four different peaks. These peaks analyzed by high performance anion-exchange with pulsed amperometric detection and size exclusion chromatography with triple detector array, ion chromatography and by mono- or two-dimensional nuclear magnetic resonance revealed a heterogeneous composition of both glycosaminoglycans in terms of sulfation grade and molecular weight. High molecular weight species (>100 KDa) were also present in the samples that counted for chains still partially linked to a proteoglycan core. - Highlights: • A multi-analytical approach was set up, for the first time, for the determination of the residual keratan sulfate

A multi-analytical approach to better assess the keratan sulfate contamination in animal origin chondroitin sulfate

International Nuclear Information System (INIS)

Restaino, Odile Francesca; Finamore, Rosario; Diana, Paola; Marseglia, Mariacarmela; Vitiello, Mario; Casillo, Angela; Bedini, Emiliano; Parrilli, Michelangelo

2017-01-01

Chondroitin sulfate is a glycosaminoglycan widely used as active principle of anti-osteoarthritis drugs and nutraceuticals, manufactured by extraction from animal cartilaginous tissues. During the manufacturing procedures, another glycosaminoglycan, the keratan sulfate, might be contemporarily withdrawn, thus eventually constituting a contaminant difficult to be determined because of its structural similarity. Considering the strict regulatory rules on the pureness of pharmaceutical grade chondrotin sulfate there is an urgent need and interest to determine the residual keratan sulfate with specific, sensitive and reliable methods. To pursue this aim, in this paper, for the first time, we set up a multi-analytical and preparative approach based on: i) a newly developed method by high performance anion-exchange chromatography with pulsed amperometric detection, ii) gas chromatography-mass spectrometry analyses, iii) size exclusion chromatography analyses coupled with triple detector array module and on iv) strong anion exchange chromatography separation. Varied KS percentages, in the range from 0.1 to 19.0% (w/w), were determined in seven pharmacopeia and commercial standards and nine commercial samples of different animal origin and manufacturers. Strong anion exchange chromatography profiles of the samples showed three or four different peaks. These peaks analyzed by high performance anion-exchange with pulsed amperometric detection and size exclusion chromatography with triple detector array, ion chromatography and by mono- or two-dimensional nuclear magnetic resonance revealed a heterogeneous composition of both glycosaminoglycans in terms of sulfation grade and molecular weight. High molecular weight species (>100 KDa) were also present in the samples that counted for chains still partially linked to a proteoglycan core. - Highlights: • A multi-analytical approach was set up, for the first time, for the determination of the residual keratan sulfate

Glycosaminoglycan synthesis by human chondrosarcoma

International Nuclear Information System (INIS)

Thonar, E.J-M.A.; Lyons, G.; Sweet, M.B.; Immelman, A.R.

1979-01-01

Human chondrosarcoma of low-grade malignancy was cultured in the presence of 35 S-sulphate and 3 H-glucosamine. The glycosaminoglycans isolated were fractioned on Ecteola cellulose and electrophoresed on cellulose acetate membranes before and after treatment with chondroitinase AC or Streptomyces hyaluronidase. The results demonstrated the in vitro synthesis of hyaluronate, chondroitin sulphate and keratan sulphate. The presence of keratan sulphate of large average chain length (approximately equal to 15 monosaccharides) supports the contention that chain length of keratan sulphate is inversely proportional to the degree of malignancy

Effects of sulfate deprivation on the production of chondroitin/dermatan sulfate by cultures of skin fibroblasts from normal and diabetic individuals

International Nuclear Information System (INIS)

Silbert, C.K.; Humphries, D.E.; Palmer, M.E.; Silbert, J.E.

1991-01-01

Human skin fibroblast monolayer cultures from two normal men, three Type I diabetic men, and one Type I diabetic woman were incubated with [3H]glucosamine in the presence of diminished concentrations of sulfate. Although total synthesis of [3H]chondroitin/dermatan glycosaminoglycans varied somewhat between cell lines, glycosaminoglycan production was not affected within any line when sulfate levels were decreased from 0.3 mM to 0.06 mM to 0.01 mM to 0 added sulfate. Lowering of sulfate concentrations resulted in diminished sulfation of chondroitin/dermatan in a progressive manner, so that overall sulfation dropped to as low as 19% for one of the lines. Sulfation of chondroitin to form chondroitin 4-sulfate and chondroitin 6-sulfate was progressively and equally affected by decreasing the sulfate concentration in the culture medium. However, sulfation to form dermatan sulfate was preserved to a greater degree, so that the relative proportion of dermatan sulfate to chondroitin sulfate increased. Essentially all the nonsulfated residues were susceptible to chondroitin AC lyase, indicating that little epimerization of glucuronic acid residues to iduronic acid had occurred in the absence of sulfation. These results confirm the previously described dependency of glucuronic/iduronic epimerization on sulfation, and indicate that sulfation of the iduronic acid-containing disaccharide residues of dermatan can take place with sulfate concentrations lower than those needed for 6-sulfation and 4-sulfation of the glucuronic acid-containing disaccharide residues of chondroitin. There were considerable differences among the six fibroblast lines in susceptibility to low sulfate medium and in the proportion of chondroitin 6-sulfate, chondroitin 4-sulfate, and dermatan sulfate. However, there was no pattern of differences between normals and diabetics

Presynaptic proteoglycans: sweet organizers of synapse development.

Science.gov (United States)

Song, Yoo Sung; Kim, Eunjoon

2013-08-21

Synaptic adhesion molecules control neuronal synapse development. In this issue of Neuron, Siddiqui et al. (2013) and de Wit et al. (2013) demonstrate that LRRTM4, a postsynaptic adhesion molecule, trans-synaptically interacts with presynaptic heparan sulfate proteoglycans (HSPGs) to promote synapse development. Copyright © 2013 Elsevier Inc. All rights reserved.

Sulphate solubility and sulphate diffusion in oxide glasses: implications for the containment of sulphate-bearing nuclear wastes; Solubilite et cinetiques de diffusion des sulfates dans differents verres d'oxydes: application au conditionnement des dechets nucleaires sulfates

Energy Technology Data Exchange (ETDEWEB)

Lenoir, M.

2009-09-15

The thesis deals with sulphate solubility and sulphate diffusion in oxide glasses, in order to control sulphate incorporation and sulphate volatilization in nuclear waste glasses. It was conducted on simplified compositions, in the SiO{sub 2}-B{sub 2}O{sub 3}-R{sub 2}O (R = Li, Na, K, Cs), SiO{sub 2}-B{sub 2}O{sub 3}-BaO and V{sub 2}O{sub 5}-B{sub 2}O{sub 3}-BaO systems. These compositions allowed us to study the influence of the nature of network-modifying ions (Li{sup +}, Na{sup +}, K{sup +}, Cs{sup +} or Ba{sup 2+}) and also of former elements (Si, B, V), on structure and properties of glasses. Sulphate volatility is studied in sodium borosilicate melts using an innovative technique of sulphate quantitation with Raman spectroscopy. This technique is useful to obtain kinetic curves of sulphate volatilization. The establishment of a model to fit these curves leads to the determination of diffusion coefficients of sulphate. These diffusion coefficients can thus be compared to diffusion coefficients of other species, determined by other techniques and presented in the literature. They are also linked to diffusion coefficients in relation with the viscosity of the melts. Concerning sulphate solubility in glasses, it depends on glass composition and on the nature of sulphate incorporated. Sulphate incorporation in alkali borosilicate glasses leads to the formation of a sulphate layer floating on top of the melt. Sulphate incorporation in barium borosilicate and boro-vanadate glasses leads to the crystallization of sulphate species inside the vitreous matrix. Moreover, sulphate solubility is higher in these glasses than in alkali borosilicates. Finally, exchanges between cations present in glasses and cations present in the sulphate phase are also studied. (author)

Characterization of a dermatan sulfate proteoglycan synthesized by murine parietal yolk sac (PYS-2) cells

DEFF Research Database (Denmark)

Couchman, J R; Woods, A; Höök, M

1985-01-01

-polyacrylamide gel electrophoresis of chondroitinase ABC-treated 125I-labeled proteoglycan reveals two polypeptides with molecular weights of 34,000 and 27,000. Results from papain digestion of the proteoglycan suggest that most of the polysaccharide chains are clustered at a papain-resistant segment of the core...

Dermatan Sulfate Epimerase 1-Deficient Mice Have Reduced Content and Changed Distribution of Iduronic Acids in Dermatan Sulfate and an Altered Collagen Structure in Skin

DEFF Research Database (Denmark)

Maccarana, M.; Kalamajski, S.; Kongsgaard, M.

2009-01-01

Dermatan sulfate epimerase 1 (DS-epi1) and DS-epi2 convert glucuronic acid to iduronic acid in chondroitin/dermatan sulfate biosynthesis. Here we report on the generation of DS-epi1-null mice and the resulting alterations in the chondroitin/dermatan polysaccharide chains. The numbers of long blocks......-derived chains. DS-epi1-deficient mice are smaller than their wild-type littermates but otherwise have no gross macroscopic alterations. The lack of DS-epi1 affects the chondroitin/dermatan sulfate in many proteoglycans, and the consequences for skin collagen structure were initially analyzed. We found...... that the skin collagen architecture was altered, and electron microscopy showed that the DS-epi1-null fibrils have a larger diameter than the wild-type fibrils. The altered chondroitin/dermatan sulfate chains carried by decorin in skin are likely to affect collagen fibril formation and reduce the tensile...

Modulators of axonal growth and guidance at the brain midline with special reference to glial heparan sulfate proteoglycans

Directory of Open Access Journals (Sweden)

CAVALCANTE LENY A.

2002-01-01

Full Text Available Bilaterally symmetric organisms need to exchange information between the left and right sides of their bodies to integrate sensory input and to coordinate motor control. Thus, an important choice point for developing axons is the Central Nervous System (CNS midline. Crossing of this choice point is influenced by highly conserved, soluble or membrane-bound molecules such as the L1 subfamily, laminin, netrins, slits, semaphorins, Eph-receptors and ephrins, etc. Furthermore, there is much circumstantial evidence for a role of proteoglycans (PGs or their glycosaminoglycan (GAG moieties on axonal growth and guidance, most of which was derived from simplified models. A model of intermediate complexity is that of cocultures of young neurons and astroglial carpets (confluent cultures obtained from medial and lateral sectors of the embryonic rodent midbrain soon after formation of its commissures. Neurite production in these cocultures reveals that, irrespective of the previous location of neurons in the midbrain, medial astrocytes exerted an inhibitory or non-permissive effect on neuritic growth that was correlated to a higher content of both heparan and chondroitin sulfates (HS and CS. Treatment with GAG lyases shows minor effects of CS and discloses a major inhibitory or non-permissive role for HS. The results are discussed in terms of available knowledge on the binding of HSPGs to interative proteins and underscore the importance of understanding glial polysaccharide arrays in addition to its protein complement for a better understanding of neuron-glial interactions.

Removal heavy metals and sulphate from waste waters by sulphate-reducing bacteria

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Kušnierová Mária

2000-09-01

Full Text Available This article is devoted to the process of bacterial sulphate reduction, which is used to removal of heavy metals and sulphate ions from waste waters.The life of animals and plants depends on the existence of microscopic organisms – microorganisms (MO, which play an important role in cycle changes of biogenic elements on the earth. The sulphur cycle in the nature is considered as one of the oldest and most significant biological systems (Fig. 1. The sulphate-reducing bacteria (SRB miss the assimilatory part of the cycle and produce sulphides. The microbial population of this dissimilatory part is called “sulfuretum”. The SRB can be found in anaerobic mud and sediments of freshwater, thermal or non-thermal sulphur springs, mining waters from sulphide deposits, oil deposits, sea and ocean beds, and in the gastrointestinal tract of man and animals. The SRB represent a group of chemoorganotrophic, strictly anaerobic and gramnegative bacteria, which exhibit a great morphological and physiological diversity. Despite of their considerable morphological variety, they have one property in common, which is the ability to utilise preferentially sulphates (occasionally sulphites, thiosulphates, tetrathionates as electron acceptors, which are reduced to sulphides, during anaerobic respiration. The electron donors in these processes are simple organic compounds as lactate, malate, etc.,(heterotrophically reduction or gaseous hydrogen (autotrophically reduction. SRB can produce a considerable amount of hydrogen sulphide, which reacts easily in aqueous solution with the cations of heavy metals, forming metal sulphides that have low solubility. The bacterial sulphate reduction can be used for the treatment of acid mine drainage waters, which is considered to be the major problem associated with mining activities.In order to remove heavy metals from waste waters, e.g., from galvanizing plants, mine waters (Smolnik, Šobov locality and metallurgic plants (works

Removal of Sulphate and Manganese on Synthetic Wastewater in Sulphate Reducing Bioreactor Using Indonesian Natural Zeolite

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Endah Retnaningrum

2017-07-01

Full Text Available The present research was conducted to investigate sulphate and manganese removal from synthetic wastewater. The continuous laboratory scale of down-flow fluidized-bed reactor (DFBR using sulphate reducing bacteria (SRB consortium and Indonesian natural zeolite as a bacterial support material was designed. At 9 days operation, maximum sulphate and manganese removal was observed to be 23% and 15.4%, respectively. The pH values were also changed to neutral. The population of SRB increased which effect on the raising of their activity for removing sulphate and manganese. Using the scanning electronic microscopy (SEM, it was observed that natural zeolite possesses excellent physical characteristics as a bacterial support material in DFBR. The imaging SEM result of SRB consortium on zeolite surface clearly showed the developed SRB biofilm on that particle. Analysis result of EDX confirmed that manganese was precipitated as manganese–sulfides.

Effectivness of dexametasone vs. Magnesium sulphate in postoperative analgesia: Dexametasone vs. Magnesium sulphate

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Dautaj Brikena

2016-01-01

Full Text Available Introduction: Preoperative use of additive substances may be very helpful in perioperative acute pain management. Intravenous administration of dexametasone in preoperative period prevents postoperative nausea and vomiting but also provides better pain relief. It is also well known that magnesium sulphate (the NMDA receptor's antagonist by its central mechanism of action may be effective in postoperative pain control. Aim: The purpose of this study was to evaluate the effect of dexametasone and magnesium sulphate on postoperative pain management in patients undergoing abdominal surgery (open cholecystectomy. Methods: Seventy eight patients scheduled for elective surgery (open cholecystectomy were included in this study. This was the prospective cohort randomized placebo- controlled study. A total of 78 patients were randomized into three groups. Each group had twenty six patients. The group D, received dexametasone 0.1 mg/ kg iv 30 minutes before surgery. The group M received magnesium sulphate 3 mg/kg iv 30 minutes before surgery. The third group S was placebo group and patients in this group received saline in the same volume for each patient. For pain control after surgery all patients received tramadol 0.9-1.2 mg/kg and diklophenac 1.76 mg/kg. When necessary (VAS ≥ 7, morphine sulphate in dose 0.15 mg/kg was administred subcutaneously For treatment of emetic episodes metoclopramid 10 mg iv. was used. The patients were observed for intensity of pain measured VAS 0-10, pain relief and satisfaction with therapy, sedation, adverse events, emetic episodes and hemodynamic parameters. Results: There was no difference between groups regarding demographic data (age, gender, body weight, ASA score, comorbidity, duration of surgery and anesthesia and amount of fentanyl received during surgery. In group D 11.54% of patients received additional analgesia (morphine sulphate 0.15 mg/kg sc in the first 4 hours and 27% of patients in the first 24 hours

Selective rab11 transport and the intrinsic regenerative ability of CNS axons

Czech Academy of Sciences Publication Activity Database

Koseki, H.; Donegá, M.; Lam, B.Y.H.; Petrová, V.; van Erp, S.; Yeo, G.S.H.; Kwok, Jessica; Ffrench-Constant, Ch.; Eva, R.; Fawcett, James

2017-01-01

Roč. 6, aug (2017), e26956 ISSN 2050-084X R&D Projects: GA MŠk(CZ) EF15_003/0000419 Institutional support: RVO:68378041 Keywords : spinal-cord-injury * chondroitin sulfate proteoglycans * growth cone Subject RIV: FH - Neurology OBOR OECD: Neuroscience s (including psychophysiology Impact factor: 7.725, year: 2016

Chondroitin 4-O-Sulfotransferase Is Indispensable for Sulfation of Chondroitin and Plays an Important Role in Maintaining Normal Life Span and Oxidative Stress Responses in Nematodes*

Science.gov (United States)

Izumikawa, Tomomi; Dejima, Katsufumi; Watamoto, Yukiko; Nomura, Kazuko H.; Kanaki, Nanako; Rikitake, Marika; Tou, Mai; Murata, Daisuke; Yanagita, Eri; Kano, Ai; Mitani, Shohei; Nomura, Kazuya; Kitagawa, Hiroshi

2016-01-01

Chondroitin sulfate (CS)/chondroitin (Chn) chains are indispensable for embryonic cell division and cytokinesis in the early developmental stages in Caenorhabditis elegans and mice, whereas heparan sulfate (HS) is essential for axon guidance during nervous system development. These data indicate that the fundamental functions of CS and HS are conserved from worms to mammals and that the function of CS/Chn differs from that of HS. Although previous studies have shown that C. elegans produces HS and non-sulfated Chn, whether the organism produces CS remains unclear. Here, we demonstrate that C. elegans produces a small amount of 4-O-sulfated Chn and report the identification of C41C4.1, an orthologue of the human chondroitin 4-O-sulfotransferase gene. Loss of C41C4.1 in C. elegans resulted in a decline in 4-O-sulfation of CS and an increase in the number of sulfated units in HS. C41C4.1 deletion mutants exhibited reduced survival rates after synchronization with sodium hypochlorite. Collectively, these results show for the first time that CS glycans are present in C. elegans and that the Chn 4-O-sulfotransferase responsible for the sulfation plays an important role in protecting nematodes from oxidative stress. PMID:27645998

Basement membrane proteoglycans are of epithelial origin in rodent skin

DEFF Research Database (Denmark)

Yamane, Y; Yaoita, H; Couchman, J R

1996-01-01

. For in vivo experiments, pieces of newborn rat epidermis obtained by dispase treatment were grafted onto athymic nude mice. Three and six weeks after grafting, immunofluorescence analysis of the grafted skin was carried out, using monoclonal antibodies specific for rat basement membrane chondroitin sulfate...

Spatiotemporal expression of chondroitin sulfate sulfotransferases in the postnatal developing mouse cerebellum.

Science.gov (United States)

Ishii, Maki; Maeda, Nobuaki

2008-08-01

Chondroitin sulfate (CS) proteoglycans are major components of the cell surface and the extracellular matrix in the developing brain and bind to various proteins via CS chains in a CS structure-dependent manner. This study demonstrated the expression pattern of three CS sulfotransferase genes, dermatan 4-O-sulfotransferase (D4ST), uronyl 2-O-sulfotransferase (UST), and N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase (GalNAc4S-6ST), in the mouse postnatal cerebellum. These sulfotransferases are responsible for the biosynthesis of oversulfated structures in CS chains such as B, D, and E units, which constitute the binding sites for various heparin-binding proteins. Real-time reverse transcription-polymerase chain reaction analysis indicated that the expression of UST increased remarkably during cerebellar development. The amounts of B and D units, which are generated by UST activity, in the cerebellar CS chains also increased during development. In contrast, the expression of GalNAc4S-6ST and its biosynthetic product, E unit, decreased during postnatal development. In situ hybridization experiments revealed the levels of UST and GalNAc4S-6ST mRNAs to correlate inversely in many cells including Purkinje cells, granule cells in the external granular layer, and inhibitory interneurons. In these neurons, the expression of UST increased and that of GalNAc4S-6ST decreased during development and/or maturation. D4ST was also expressed by many neurons, but its expression was not simply correlated with development, which might contribute to the diversification of CS structures expressed by distinct neurons. These results suggest that the CS structures of various cerebellar neurons change during development and such changes of CS are involved in the regulation of various signaling pathways.

Analysis of oversulfation in biglycan chondroitin/dermatan sulfate oligosaccharides by chip-based nanoelectrospray ionization multistage mass spectrometry.

Science.gov (United States)

Flangea, Corina; Sisu, Eugen; Seidler, Daniela G; Zamfir, Alina D

2012-01-15

Biglycan (BGN) is a small proteoglycan that consists of a protein core containing leucine-rich repeat regions and two glycosaminoglycan (GAG) chains of either chondroitin sulfate (CS) or dermatan sulfate (DS) type. The development of novel, highly efficient analytical methods for structural identification of BGN-derived CS/DS motifs, possibly implicated in biological events, is currently the focus of research. In this work, an improved analytical method based on fully automated chip-nanoelectrospray ionization (nanoESI) in conjunction with high-capacity ion trap (HCT) multistage mass spectrometry (MS) by collision-induced dissociation (CID) was for the first time applied to BGN CS/DS oligosaccharide analysis. The CS/DS chains were released from transfected 293 BGN by β-elimination. The chain was digested with AC I lyase, and the resulting mixture was purified and subsequently separated by size exclusion chromatography (SEC). Di- and tetrasaccharide fractions were pooled and characterized in detail using the developed chip-nanoESI protocol. The chip-nanoESI MS profile in the negative ion mode revealed the presence of under-, regularly, and oversulfated species in both di- and tetrasaccharide fractions. CID MS(2)-MS(3) yielded sequence patterns consistent with unusual oversulfated 4,5-Δ-GlcA(2S)-GalNAc(4S) and 4,5-Δ-GlcA(2S)-GalNAc(6S)-IdoA(2S)-GalNAc(6S) motifs. Copyright © 2011 Elsevier Inc. All rights reserved.

Utilization of Glycosaminoglycans/Proteoglycans as Carriers for Targeted Therapy Delivery

Science.gov (United States)

Misra, Suniti; Hascall, Vincent C.; Atanelishvili, Ilia; Moreno Rodriguez, Ricardo; Markwald, Roger R.; Ghatak, Shibnath

2015-01-01

The outcome of patients with cancer has improved significantly in the past decade with the incorporation of drugs targeting cell surface adhesive receptors, receptor tyrosine kinases, and modulation of several molecules of extracellular matrices (ECMs), the complex composite of collagens, glycoproteins, proteoglycans, and glycosaminoglycans that dictates tissue architecture. Cancer tissue invasive processes progress by various oncogenic strategies, including interfering with ECM molecules and their interactions with invasive cells. In this review, we describe how the ECM components, proteoglycans and glycosaminoglycans, influence tumor cell signaling. In particular this review describes how the glycosaminoglycan hyaluronan (HA) and its major receptor CD44 impact invasive behavior of tumor cells, and provides useful insight when designing new therapeutic strategies in the treatment of cancer. PMID:26448753

Extraction of sulphates by long chain amines

International Nuclear Information System (INIS)

Boirie, Ch.

1959-05-01

The extraction of sulphuric acid by long chain amines in organic solution has been studied with a view to determining the value of the stability constants of the amine sulphates and bi-sulphates formed. We have concentrated chiefly on uranium sulphate and thorium sulphate. The formulae of the complexes extractable with amines have been established, as well as the corresponding dissociation constants. We have observed that for uranium sulphate the formula of the complex depends only on the nature of the amine, whereas for thorium this formula varies with the amine structure. From the formulae determined and the value of the constants calculated, we have been able to establish the best conditions for uranium and thorium extraction and also for a separation of these two elements. Finally we propose an application of this study to the determination of uranium in ores, where the separation of uranium by this method is particularly easy and complete. (author) [fr

Reactivating the extracellular matrix synthesis of sulfated glycosaminoglycans and proteoglycans to improve the human skin aspect and its mechanical properties

Directory of Open Access Journals (Sweden)

Chajra H

2016-12-01

, and versican and a stimulation of their respective sGAGs, such as chondroitin sulfate and heparan sulfate, were found on skin explants. The biosynthesis of macromolecules seems to be correlated at the microscopic level to a better organization and quality of the dermis, with collagen fibrils having homogenous diameters. The dermis seems to be compacted as observed on images obtained by two-photon microscopy and ultrasound imaging. At the macroscopic level, this dermis organization shows a smoothed profile similar to a younger skin, with improved mechanical properties such as firmess. Conclusion: The obtained results demonstrate that the defined cosmetic composition induces the synthesis of sGAGs and proteoglycans, which contributes to the overall dermal reorganization. This activity in the dermis in turn impacts the surface and mechanical properties of the skin. Keywords: cosmetic composition, decorin, dermis, polysaccharide

Sulphate deposition by precipitation into Lake Ontario

Energy Technology Data Exchange (ETDEWEB)

Shaw, R W; Whelpdale, D M

1973-01-01

Measurements of sulphate concentration in precipitation from individual snow storms of several hours duration in the western Lake Ontario region indicate that approximately 9-66 mg/M/sub 2/ of SO/sub 4//sup 2 -/ is being deposited into the lake per storm. This amount is up to several times more than daily average values over long periods found by other workers. Using a mean sulphate concentration of 4 mg/l and an annual accumulation of precipitation of 760 mm, the yearly sulphate deposition by precipitation is about 0.1% of the total mass of sulphate in the lake; however, more significantly, it is of the same order of magnitude as that discharged directly into the lake by industry.

A potential role for chondroitin sulfate/dermatan sulfate in arm regeneration in Amphiura filiformis.

Science.gov (United States)

Ramachandra, Rashmi; Namburi, Ramesh B; Dupont, Sam T; Ortega-Martinez, Olga; van Kuppevelt, Toin H; Lindahl, Ulf; Spillmann, Dorothe

2017-05-01

Glycosaminoglycans (GAGs), such as chondroitin sulfate (CS) and dermatan sulfate (DS) from various vertebrate and invertebrate sources are known to be involved in diverse cellular mechanisms during repair and regenerative processes. Recently, we have identified CS/DS as the major GAG in the brittlestar Amphiura filiformis, with high proportions of di- and tri-O-sulfated disaccharide units. As this echinoderm is known for its exceptional regeneration capacity, we aimed to explore the role of these GAG chains during A. filiformis arm regeneration. Analysis of CS/DS chains during the regeneration process revealed an increase in the proportion of the tri-O-sulfated disaccharides. Conversely, treatment of A. filiformis with sodium chlorate, a potent inhibitor of sulfation reactions in GAG biosynthesis, resulted in a significant reduction in arm growth rates with total inhibition at concentrations higher than 5 mM. Differentiation was less impacted by sodium chlorate exposure or even slightly increased at 1-2 mM. Based on the structural changes observed during arm regeneration we identified chondroitin synthase, chondroitin-4-O-sulfotransferase 2 and dermatan-4-O-sulfotransferase as candidate genes and sought to correlate their expression with the expression of the A. filiformis orthologue of bone morphogenetic factors, AfBMP2/4. Quantitative amplification by real-time PCR indicated increased expression of chondroitin synthase and chondroitin-4-O-sulfotransferase 2, with a corresponding increase in AfBMP2/4 during regeneration relative to nonregenerating controls. Our findings suggest that proper sulfation of GAGs is important for A. filiformis arm regeneration and that these molecules may participate in mechanisms controlling cell proliferation. © The Author 2017. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Mortar and concrete based on calcium sulphate binders

NARCIS (Netherlands)

Bakker, J.J.F.; Brouwers, H.J.H.; Fischer, H.B

2006-01-01

In this study both hemi-hydrate and anhydrite are tested as calcium sulphate binders for structural mortar and concrete. The advantage of using calcium sulphates instead of cement as a binder is the fact that the production of calcium sulphate is more environmental friendly than that of cement. For

Mammalian tissue distribution of a large heparan sulfate proteoglycan detected by monoclonal antibodies

DEFF Research Database (Denmark)

Couchman, J R; Ljubimov, A V

1989-01-01

muscle, endothelia, peripheral nerve fibers and epithelia so far examined. In addition, two of the monoclonal antibodies show cross-species reactivity, staining bovine and human basement membranes, and immunoprecipitating proteoglycans from human endothelial cell cultures. These antibodies do not......A panel of nine monoclonal antibodies has been characterized, all of which have reactivity with the core protein of a large heparan sulfate proteoglycan derived from the murine EHS tumor matrix. These rat monoclonal antibodies stained mouse basement membranes intensely, including those of all...

Physicochemical and in vitro deposition properties of salbutamol sulphate/ipratropium bromide and salbutamol sulphate/excipient spray dried mixtures for use in dry powder inhalers.

Science.gov (United States)

Corrigan, Deirdre O; Corrigan, Owen I; Healy, Anne Marie

2006-09-28

The physicochemical and aerodynamic properties of spray dried powders of the drug/drug mixture salbutamol sulphate/ipratropium bromide were investigated. The in vitro deposition properties of spray dried salbutamol sulphate and the spray dried drug/excipient mixtures salbutamol sulphate/lactose and salbutamol sulphate/PEG were also determined. Spray drying ipratropium bromide monohydrate resulted in a crystalline material from both aqueous and ethanolic solution. The product spray dried from aqueous solution consisted mainly of ipratropium bromide anhydrous. There was evidence of the presence of another polymorphic form of ipratropium bromide. When spray dried from ethanolic solution the physicochemical characterisation suggested the presence of an ipratropium bromide solvate with some anhydrous ipratropium bromide. Co-spray drying salbutamol sulphate with ipratropium bromide resulted in amorphous composites, regardless of solvent used. Particles were spherical and of a size suitable for inhalation. Twin impinger studies showed an increase in the fine particle fraction (FPF) of spray dried salbutamol sulphate compared to micronised salbutamol sulphate. Co-spray dried salbutamol sulphate:ipratropium bromide 10:1 and 5:1 systems also showed an increase in FPF compared to micronised salbutamol sulphate. Most co-spray dried salbutamol sulphate/excipient systems investigated demonstrated FPFs greater than that of micronised drug alone. The exceptions to this were systems containing PEG 4000 20% or PEG 20,000 40% both of which had FPFs not significantly different from micronised salbutamol sulphate. These two systems were crystalline unlike most of the other spray dried composites examined which were amorphous in nature.

Antiaging effects of a novel facial serum containing L-ascorbic acid, proteoglycans, and proteoglycan-stimulating tripeptide: ex vivo skin explant studies and in vivo clinical studies in women

Directory of Open Access Journals (Sweden)

Garre A

2018-05-01

Full Text Available Aurora Garre,1 Mridvika Narda,1 Palmira Valderas-Martinez,1 Jaime Piquero,2 Corinne Granger1 1Innovation and Development, ISDIN SA, Barcelona, Spain; 2Dermik Clinic, Barcelona, Spain Background: With age, decreasing dermal levels of proteoglycans, collagen, and elastin lead to the appearance of aged skin. Oxidation, largely driven by environmental factors, plays a central role.Aim: The aim of this study was to assess the antiaging efficacy of a topical serum containing l-ascorbic acid, soluble proteoglycans, low molecular weight hyaluronic acid, and a tripeptide in ex vivo and in vivo clinical studies.Methods: Photoaging and photo-oxidative damage were induced in human skin explants by artificial solar radiation. Markers of oxidative stress – reactive oxygen species (ROS, total glutathione (GSH, and cyclobutane pyrimidine dimers (CPDs – were measured in serum-treated explants and untreated controls. Chronological aging was simulated using hydrocortisone. In both ex vivo studies, collagen, elastin, and proteoglycans were determined as measures of dermal matrix degradation. In women aged 21–67 years, hydration was measured up to 24 hours after a single application of serum, using Corneometer and hygrometer. Subjects’ perceptions of efficacy and acceptability were assessed via questionnaire after once-daily serum application for 4 weeks. Studies were performed under the supervision of a dermatologist.Results: In the photoaging study, irradiation induced changes in ROS, CPD, GSH, collagen, and elastin levels; these changes were reversed by topical serum application. The serum also protected against hydrocortisone-induced reduction in collagen, elastin, and proteoglycan levels, which were significantly higher in the serum-treated group vs untreated hydrocortisone-control explants. In clinical studies, serum application significantly increased skin moisture for 6 hours. Healthy volunteers perceived the product as efficient in making the

Sulphate reduction and vertical distribution of sulphate-reducing bacteria quantified by rRNA slot-blot hybridization in a coastal marine sediment

DEFF Research Database (Denmark)

Sahm, K.; MacGregor, BJ; Jørgensen, BB

1999-01-01

In the past, enumeration of sulphate-reducing bacteria (SRB) by cultivation-based methods generally contradicted measurements of sulphate reduction, suggesting unrealistically high respiration rates per cell. Here, we report evidence that quantification of SRB rRNA by slot-blot hybridization......, directly above the sulphate reduction maximum. Cell numbers calculated by converting the relative contribution of SRB rRNA to the percentage of DAPI-stained cells indicated a population size for SRB of 2.4-6.1 x 10(8) cells cm(-3) wet sediment. Cellular sulphate reduction rates calculated on the basis...

Chondroitin 4-O-Sulfotransferase Is Indispensable for Sulfation of Chondroitin and Plays an Important Role in Maintaining Normal Life Span and Oxidative Stress Responses in Nematodes.

Science.gov (United States)

Izumikawa, Tomomi; Dejima, Katsufumi; Watamoto, Yukiko; Nomura, Kazuko H; Kanaki, Nanako; Rikitake, Marika; Tou, Mai; Murata, Daisuke; Yanagita, Eri; Kano, Ai; Mitani, Shohei; Nomura, Kazuya; Kitagawa, Hiroshi

2016-10-28

Chondroitin sulfate (CS)/chondroitin (Chn) chains are indispensable for embryonic cell division and cytokinesis in the early developmental stages in Caenorhabditis elegans and mice, whereas heparan sulfate (HS) is essential for axon guidance during nervous system development. These data indicate that the fundamental functions of CS and HS are conserved from worms to mammals and that the function of CS/Chn differs from that of HS. Although previous studies have shown that C. elegans produces HS and non-sulfated Chn, whether the organism produces CS remains unclear. Here, we demonstrate that C. elegans produces a small amount of 4-O-sulfated Chn and report the identification of C41C4.1, an orthologue of the human chondroitin 4-O-sulfotransferase gene. Loss of C41C4.1 in C. elegans resulted in a decline in 4-O-sulfation of CS and an increase in the number of sulfated units in HS. C41C4.1 deletion mutants exhibited reduced survival rates after synchronization with sodium hypochlorite. Collectively, these results show for the first time that CS glycans are present in C. elegans and that the Chn 4-O-sulfotransferase responsible for the sulfation plays an important role in protecting nematodes from oxidative stress. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

High-oleic canola oil consumption enriches LDL particle cholesteryl oleate content and reduces LDL proteoglycan binding in humans.

Science.gov (United States)

Jones, Peter J H; MacKay, Dylan S; Senanayake, Vijitha K; Pu, Shuaihua; Jenkins, David J A; Connelly, Philip W; Lamarche, Benoît; Couture, Patrick; Kris-Etherton, Penny M; West, Sheila G; Liu, Xiaoran; Fleming, Jennifer A; Hantgan, Roy R; Rudel, Lawrence L

2015-02-01

Oleic acid consumption is considered cardio-protective according to studies conducted examining effects of the Mediterranean diet. However, animal models have shown that oleic acid consumption increases LDL particle cholesteryl oleate content which is associated with increased LDL-proteoglycan binding and atherosclerosis. The objective was to examine effects of varying oleic, linoleic and docosahexaenoic acid consumption on human LDL-proteoglycan binding in a non-random subset of the Canola Oil Multi-center Intervention Trial (COMIT) participants. COMIT employed a randomized, double-blind, five-period, cross-over trial design. Three of the treatment oil diets: 1) a blend of corn/safflower oil (25:75); 2) high oleic canola oil; and 3) DHA-enriched high oleic canola oil were selected for analysis of LDL-proteoglycan binding in 50 participants exhibiting good compliance. LDL particles were isolated from frozen plasma by gel filtration chromatography and LDL cholesteryl esters quantified by mass-spectrometry. LDL-proteoglycan binding was assessed using surface plasmon resonance. LDL particle cholesterol ester fatty acid composition was sensitive to the treatment fatty acid compositions, with the main fatty acids in the treatments increasing in the LDL cholesterol esters. The corn/safflower oil and high-oleic canola oil diets lowered LDL-proteoglycan binding relative to their baseline values (p = 0.0005 and p = 0.0012, respectively). At endpoint, high-oleic canola oil feeding resulted in lower LDL-proteoglycan binding than corn/safflower oil (p = 0.0243) and DHA-enriched high oleic canola oil (p = 0.0249), although high-oleic canola oil had the lowest binding at baseline (p = 0.0344). Our findings suggest that high-oleic canola oil consumption in humans increases cholesteryl oleate percentage in LDL, but in a manner not associated with a rise in LDL-proteoglycan binding. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Large proteoglycan complexes and disturbed collagen architecture in the corneal extracellular matrix of mucopolysaccharidosis type VII (Sly syndrome).

Science.gov (United States)

Young, Robert D; Liskova, Petra; Pinali, Christian; Palka, Barbara P; Palos, Michalis; Jirsova, Katerina; Hrdlickova, Enkela; Tesarova, Marketa; Elleder, Milan; Zeman, Jiri; Meek, Keith M; Knupp, Carlo; Quantock, Andrew J

2011-08-24

Deficiencies in enzymes involved in proteoglycan (PG) turnover underlie a number of rare mucopolysaccharidoses (MPS), investigations of which can considerably aid understanding of the roles of PGs in corneal matrix biology. Here, the authors analyze novel pathologic changes in MPS VII (Sly syndrome) to determine the nature of PG-collagen associations in stromal ultrastructure. Transmission electron microscopy and electron tomography were used to investigate PG-collagen architectures and interactions in a cornea obtained at keratoplasty from a 22-year-old man with MPS VII, which was caused by a compound heterozygous mutation in the GUSB gene. Transmission electron microscopy showed atypical morphology of the epithelial basement membrane and Bowman's layer in MPS VII. Keratocytes were packed with cytoplasmic vacuoles containing abnormal glycosaminoglycan (GAG) material, and collagen fibrils were thinner than in normal cornea and varied considerably throughout anterior (14-32 nm), mid (13-42 nm), and posterior (17-39 nm) regions of the MPS VII stroma. PGs viewed in three dimensions were striking in appearance in that they were significantly larger than PGs in normal cornea and formed highly extended linkages with multiple collagen fibrils. Cellular changes in the MPS VII cornea resemble those in other MPS. However, the wide range of collagen fibril diameters throughout the stroma and the extensive matrix presence of supranormal-sized PG structures appear to be unique features of this disorder. The findings suggest that the accumulation of stromal chondroitin-, dermatan-, and heparan-sulfate glycosaminoglycans in the absence of β-glucuronidase-mediated degradation can modulate collagen fibrillogenesis.

Sulphate reduction and vertical distribution of sulphate-reducing bacteria quantified by rRNA slot-blot hybridization in a coastal marine sediment

DEFF Research Database (Denmark)

Sahm, K.; MacGregor, BJ; Jørgensen, BB

1999-01-01

In the past, enumeration of sulphate-reducing bacteria (SRB) by cultivation-based methods generally contradicted measurements of sulphate reduction, suggesting unrealistically high respiration rates per cell. Here, we report evidence that quantification of SRB rRNA by slot-blot hybridization...... between 18% and 25% to the prokaryotic rRNA pool. The dominant SRB were related to complete oxidizing genera (Desulphococcus, Desulphosarcina and Desulphobacterium), while Desulpho-bacter could not be detected. The vertical profile and quantity of rRNA from SRB was compared with sulphate reduction rates......, directly above the sulphate reduction maximum. Cell numbers calculated by converting the relative contribution of SRB rRNA to the percentage of DAPI-stained cells indicated a population size for SRB of 2.4-6.1 x 10(8) cells cm(-3) wet sediment. Cellular sulphate reduction rates calculated on the basis...

Identification of proteinaceous material in the bone of the dinosaur Iguanodon.

Science.gov (United States)

Embery, Graham; Milner, Angela C; Waddington, Rachel J; Hall, Rachel C; Langley, Martin S; Milan, Anna M

2003-01-01

This study has directed attention at the search for bone-related proteins in an extract of demineralized rib bone of the 120 mya Iguanodon. The inner compact bone was demineralized and the GuCl extract resolved into 11 fractions using anion exchange chromatography, which all contained silver-reactive proteins with various amino acid profiles. Two specific fractions, iv and xi, revealed characteristics typical of contemporary phosphoproteins and proteoglycans, respectively. Fraction iv, 43-57 kDa, contained a high ratio of aspartate and serine, although no phosphate was discernable. Fraction xi contained a band of 41-47 kDa and was rich in chondroitin sulphate and hyaluronan. In addition an early eluting fraction was immunoreactive with an antibody against osteocalcin. A cancellous bone fraction from the same bone sample was also analyzed using N-terminal sequencing and revealed potential similarities with cystatin. While we do not claim to have identified the presence of intact proteins, this study has value in demonstrating that extruded extracellular matrix is protected by its capacity to induce mineralization, which subsequently is important in conserving detectable protein products in ancient skeletal tissues.

Is chondroitin sulfate responsible for the biological effects attributed to the GC protein-derived Macrophage Activating Factor (GcMAF)?

Science.gov (United States)

Ruggiero, Marco; Reinwald, Heinz; Pacini, Stefania

2016-09-01

We hypothesize that a plasma glycosaminoglycan, chondroitin sulfate, may be responsible for the biological and clinical effects attributed to the Gc protein-derived Macrophage Activating Factor (GcMAF), a protein that is extracted from human blood. Thus, Gc protein binds chondroitin sulfate on the cell surface and such an interaction may occur also in blood, colostrum and milk. This interpretation would solve the inconsistencies encountered in explaining the effects of GcMAF in vitro and in vivo. According to our model, the Gc protein or the GcMAF bind to chondroitin sulfate both on the cell surface and in bodily fluids, and the resulting multimolecular complexes, under the form of oligomers trigger a transmembrane signal or, alternatively, are internalized and convey the signal directly to the nucleus thus eliciting the diverse biological effects observed for both GcMAF and chondroitin sulfate. Copyright © 2016 Elsevier Ltd. All rights reserved.

Sulphate rocks as an arena for karst development

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Andrejchuk V.

1996-01-01

Full Text Available The rocks in which karst systems develop are most commonly composed of carbonate sulphate and chloride minerals. The sulphate minerals are quite numerous, but only gypsum and anhydrite form extensive masses in sedimentary sequences. Other minerals, which represent sulphates of K, Mg and Na, normally occur as minor beds (0.1-5.0 m, or as inclusions associated with chloride rocks. However some minerals precipitated in salt-generating basins, such as mirabilite and glauberite (typically formed in the Kara-Bogaz-Gol Gulf, salt lakes of Siberia and in China, form sequences up to 5-10 m thick where karst may develop. Due to the very high solubility of Na -sulphates, karst processes and features occurring in these rocks resemble salt karst. Thus, the term sulphate karst, although not strictly correct, is used mainly to indicate karst developed in gypsum and anhydrite.

A 3D-structural model of unsulfated chondroitin from high-field NMR: 4-sulfation has little effect on backbone conformation

Science.gov (United States)

Sattelle, Benedict M.; Shakeri, Javad; Roberts, Ian S.; Almond, Andrew

2010-01-01

The glycosaminoglycan chondroitin sulfate is essential in human health and disease but exactly how sulfation dictates its 3D-strucutre at the atomic level is unclear. To address this, we have purified homogenous oligosaccharides of unsulfated chondroitin (with and without 15N-enrichment) and analysed them by high-field NMR to make a comparison published chondroitin sulfate and hyaluronan 3D-structures. The result is the first full assignment of the tetrasaccharide and an experimental 3D-model of the hexasaccharide (PDB code 2KQO). In common with hyaluronan, we confirm that the amide proton is not involved in strong, persistent inter-residue hydrogen bonds. However, in contrast to hyaluronan, a hydrogen bond is not inferred between the hexosamine OH-4 and the glucuronic acid O5 atoms across the β(1→3) glycosidic linkage. The unsulfated chondroitin bond geometry differs slightly from hyaluronan by rotation about the β(1→3) ψ dihedral (as previously predicted by simulation), while the β(1→4) linkage is unaffected. Furthermore, comparison shows that this glycosidic linkage geometry is similar in chondroitin-4-sulfate. We therefore hypothesise that both hexosamine OH-4 and OH-6 atoms are solvent exposed in chondroitin, explaining why it is amenable to sulfation and hyaluronan is not, and also that 4-sulfation has little effect on backbone conformation. Our conclusions exemplify the value of the 3D-model presented here and progress our understanding of glycosaminoglycan molecular properties. PMID:20022001

Sulphate reduction experiment: SURE-1

International Nuclear Information System (INIS)

Pedersen, K.; Arlinger, J.; Bengtsson, A.; Edlund, J.; Eriksson, L.; Hallbeck, L.; Johansson, J.; Paeaejaervi, A.; Rabe, L.

2013-11-01

It was previously concluded that opposing gradients of sulphate and methane, observations of 16S rDNA sequences displaying great similarity to those of anaerobic methane-oxidizing Archaea, and a peak in sulphide concentration in groundwater from a depth of 250-350 m in Olkiluoto, Finland, indicated proper conditions for methane oxidation with sulphate. In the present research (SURE-1), pressure-resistant, gas-tight circulating systems were constructed to enable the investigation of attached and unattached anaerobic microbial populations from a depth of 327 m in Olkiluoto under in situ pressure (2.4 MPa), diversity, dissolved gas, and hydrochemical conditions of groundwater station ONKPVA6. Three parallel flow cell cabinets were configured to allow observation of the influence on microbial metabolic activity of 11 mM methane, 11 mM methane plus 10 mM H 2 , or 2.1 mM O 2 plus 7.9 mM N 2 (i.e., air). The concentrations of these gases and of organic acids and carbon, sulphur chemistry, pH and E h , ATP, numbers of cultivable microorganisms, and total numbers of cells and bacteriophages were subsequently recorded under batch conditions for 105 d. The system containing H 2 and methane displayed microbial reduction of 0.7 mM sulphate to sulphide, while the system containing only methane produced 0.2 mM reduced sulphate. The system containing added air became inhibited and displayed no signs of microbial activity. Added H 2 and methane induced increasing numbers of lysogenic bacteriophages per cell. It appears possible that a microbial anaerobic methane-oxidizing process coupled to acetate formation and sulphate reduction may be ongoing in aquifers at a depth of 250-350 m in Olkiluoto, but clear evidence of such an AOM process was not obtained. (orig.)

Sulphate reduction experiment: SURE-1

Energy Technology Data Exchange (ETDEWEB)

Pedersen, K.; Arlinger, J.; Bengtsson, A.; Edlund, J.; Eriksson, L.; Hallbeck, L.; Johansson, J.; Paeaejaervi, A.; Rabe, L. [Microbial Analytics Sweden AB, Moelnlycke (Sweden)

2013-11-15

It was previously concluded that opposing gradients of sulphate and methane, observations of 16S rDNA sequences displaying great similarity to those of anaerobic methane-oxidizing Archaea, and a peak in sulphide concentration in groundwater from a depth of 250-350 m in Olkiluoto, Finland, indicated proper conditions for methane oxidation with sulphate. In the present research (SURE-1), pressure-resistant, gas-tight circulating systems were constructed to enable the investigation of attached and unattached anaerobic microbial populations from a depth of 327 m in Olkiluoto under in situ pressure (2.4 MPa), diversity, dissolved gas, and hydrochemical conditions of groundwater station ONKPVA6. Three parallel flow cell cabinets were configured to allow observation of the influence on microbial metabolic activity of 11 mM methane, 11 mM methane plus 10 mM H{sub 2}, or 2.1 mM O{sub 2} plus 7.9 mM N{sub 2} (i.e., air). The concentrations of these gases and of organic acids and carbon, sulphur chemistry, pH and E{sub h}, ATP, numbers of cultivable microorganisms, and total numbers of cells and bacteriophages were subsequently recorded under batch conditions for 105 d. The system containing H{sub 2} and methane displayed microbial reduction of 0.7 mM sulphate to sulphide, while the system containing only methane produced 0.2 mM reduced sulphate. The system containing added air became inhibited and displayed no signs of microbial activity. Added H{sub 2} and methane induced increasing numbers of lysogenic bacteriophages per cell. It appears possible that a microbial anaerobic methane-oxidizing process coupled to acetate formation and sulphate reduction may be ongoing in aquifers at a depth of 250-350 m in Olkiluoto, but clear evidence of such an AOM process was not obtained. (orig.)

Isotopic evolution of aqueous sulphate in northern Chile water

International Nuclear Information System (INIS)

Aravena, R.; Suzuki, O.; Fritz, P.; Pena, H.; Rauert, W.

1987-01-01

Full text: The extremely arid condition of northern Chile is the main obstacle to the present and future development of urban centres and economic activities in the region. The existing water resources are scarce. During the last decade, isotope techniques have been applied to investigate aspects such as the origin and residence time of the groundwater, these being extremely important for water development and water management. This paper presents 18 O and 34 S data in aqueous sulphate, as well as 18 O, 2 H, and 3 H from springs, rivers and groundwater samples of the Pampa del Tamarugal and Salar de Llamara basins. The principal aim of this study was to investigate the isotope evolution of the sulphate, the origin of the sulphur, the groundwater flow path, and the possibility of using the 18 O of the sulphate as a tracer for estimation of the residence time of the groundwater. Springs that have their recharge area in the high Altiplano (Salar del Huasco basin) show δ values between +5.0 per mille and +6.0 per mille for the 18 O and +5.0 per mille and +9.0 per mille for the 34 S isotope. Springs from lower altitude, show an isotope content between + 8.6 per mille and + 10.6 per mille for 18 O and +7.4 per mille and + 11.7 per mille for 34 S. The groundwaters in the Pampa aquifers, based on their hydrogen and oxygen isotope composition, are associated with different recharge areas. However, these waters show an isotope range for the sulphate similar to .he one of the springs, and no clear relationships are observed between isotope content, flow path and residence time. This pattern could be related to the poorly defined aquifer systems present in the Pampa. The Llamara groundwaters have a uniform isotope content and are the most enriched in the region (δ 18 O = -6.0 per mille and δ 2 H = -50 per mille). The isotope composition of their sulphate compares well with the Pampa groundwater, indicating a similar sulphate source. The isotope composition of gypsum

Mycobacterial antigens stimulate rheumatoid mononuclear cells to cartilage proteoglycan depletion

NARCIS (Netherlands)

Wilbrink, B.; Bijlsma, J. W.; Huber-Bruning, O.; van Roy, J. L.; den Otter, W.; van Eden, W.

1990-01-01

In a coculture with porcine articular cartilage explants unstimulated blood mononuclear cells (BMC) from patients with rheumatoid arthritis (RA), but not from healthy controls, induced proteoglycan depletion of dead cartilage. Specific stimulation of the RA BMC with Mycobacterium tuberculosis (MT),

Sulphate Removal from Water by Carbon Residue from Biomass Gasification: Effect of Chemical Modification Methods on Sulphate Removal Efficiency

Directory of Open Access Journals (Sweden)

Hanna Runtti

2016-02-01

Full Text Available Sulphate removal from mine water is a problem because traditional chemical precipitation does not remove all sulphates. In addition, it creates lime sediment as a secondary waste. Therefore, an inexpensive and environmental-friendly sulphate removal method is needed in addition to precipitation. In this study, carbon residues from a wood gasification process were repurposed as precursors to a suitable sorbent for SO42- ion removal. The raw material was modified using ZnCl2, BaCl2, CaCl2, FeCl3, or FeCl2. Carbon residues modified with FeCl3 were selected for further consideration because the removal efficiency toward sulphate was the highest. Batch sorption experiments were performed to evaluate the effects of the initial pH, initial SO42- ion concentration, and contact time on sulphate removal. The removal of SO42- ions using Fe-modified carbon residue was notably higher compared with unmodified carbon residue and commercially available activated carbon. The sorption data exhibited pseudo-second-order kinetics. The isotherm analysis indicated that the sorption data of Fe-modified carbon residues can be represented by the bi-Langmuir isotherm model.

Chemical sulphate removal for treatment of construction and demolition debris leachate.

Science.gov (United States)

Kijjanapanich, Pimluck; Annachhatre, Ajit P; Esposito, Giovanni; Lens, Piet N L

2014-08-01

Construction and demolition debris (CDD) is a product of construction, renovation or demolition activities. It has a high gypsum content (52.4% of total gypsum), concentrated in the CDD sand (CDDS) fraction. To comply with the posed limit of the maximum amount of sulphate present in building sand, excess sulphate needs to be removed. In order to enable reuse of CDDS, a novel treatment process is developed based on washing of the CDDS to remove most of the gypsum, and subsequent sulphate removal from the sulphate-rich CDDS leachate. This study aims to assess chemical techniques, i.e. precipitation and adsorption, for sulphate removal from the CDDS leachate. Good sulphate removal efficiencies (up to 99.9%) from the CDDS leachate can be achieved by precipitation with barium chloride (BaCl2) and lead(II) nitrate (Pb(NO3)2). Precipitation with calcium chloride (CaCl2), calcium carbonate (CaCO3) and calcium oxide (CaO) gave less efficient sulphate removal. Adsorption of sulphate to aluminium oxide (Al2O3) yielded a 50% sulphate removal efficiency, whereas iron oxide-coated sand as adsorbent gave only poor (10%) sulphate removal efficiencies.

Uranium and sulphate values from carbonate leach process

International Nuclear Information System (INIS)

Berger, B.

1983-01-01

The process concerns the recovery of uraniferous and sulphur values from liquor resulting from the attack of sulphur containing uraniferous ores by an alkaline solution of sodium carbonate and/or bicarbonate. Ammonia is introduced into the liquor to convert any HCO 3 - to CO 3 2- . The neutralised liquor from this step is then contacted with an anion exchange resin to fix the uranium and sulphate ions, leaving a liquor containing ammonia, sodium carbonate and/or bicarbonate in solution. Uranium and sulphate ions are eluted with an ammonia carbonate and/or bicarbonate solution to yield a solution of ammonium uranyl carbonate complex and ammonium sulphate. The solution is subjected to thermal treatment until a suspension of precipitated ammonium uranate and/or diuranate is obtained in a solution of the ammonium sulphate. Carbon dioxide, ammonia and water vapor are driven off. The precipitated ammonium uranate and/or diuranate is then separated from the solution of ammonium sulphate and the precipitate is calcined to yield uranium trioxide and ammonia

Extraction and determination of chondroitin sulfate from fish processing byproducts

Science.gov (United States)

Chondroitin sulfate (CS) refers to a group of sulfated glycosaminoglycan containing a chain of alternating N-acetylgalactosamine and glucuronic acid sugars. It is a major component of the extracellular matrix of cartilage and attached to proteins. CS is usually an over the counter dietary supplement...

Visualization of liver on bone scintigraphy during treatment with iron chondroitin sulfate colloid

Energy Technology Data Exchange (ETDEWEB)

Tanaka, Shigeko; Sawa, Hisashi; Fukuda, Teruo

1983-09-01

In order to see its probable cause in the incidental liver delineation on the bone scan with sup(99m)Tc-MDP, patients and experimental rabbits were intravenously injected with Blutal (iron chondroitin sulfate) at various times following the intravenous injection of sup(99m)Tc-MDP. The liver images were obtained during the early periods. An injection of Blutal-sup(99m)Tc pertechnetate mixture did not result in any appreciable hepatic delineation. Stannous chloride in the MDP kit could have enticed formation of radiocolloids in the presence of iron chondroitin sulfate. Diagnostic problems can be avoided by carefully planning the bone scintigraphy in relation to the Blutal administration. (author).

Structural Variation of Chondroitin Sulfate Chains Contributes to the Molecular Heterogeneity of Perineuronal Nets

Directory of Open Access Journals (Sweden)

Shinji Miyata

2018-02-01

Full Text Available Aggrecan, a chondroitin sulfate (CS proteoglycan, forms lattice-like extracellular matrix structures called perineuronal nets (PNNs. Neocortical PNNs primarily ensheath parvalbumin-expressing inhibitory neurons (parvalbumin, PV cells late in brain development. Emerging evidence indicates that PNNs promote the maturation of PV cells by enhancing the incorporation of homeobox protein Otx2 and regulating experience-dependent neural plasticity. Wisteria floribunda agglutinin (WFA, an N-acetylgalactosamine-specific plant lectin, binds to the CS chains of aggrecan and has been widely used to visualize PNNs. Although PNNs show substantial molecular heterogeneity, the importance of this heterogeneity in neural plasticity remains unknown. Here, in addition to WFA lectin, we used the two monoclonal antibodies Cat315 and Cat316, both of which recognize the glycan structures of aggrecan, to investigate the molecular heterogeneity of PNNs. WFA detected the highest number of PNNs in all cortical layers, whereas Cat315 and Cat316 labeled only a subset of PNNs. WFA+, Cat315+, and Cat316+ PNNs showed different laminar distributions in the adult visual cortex. WFA, Cat315 and Cat316 detected distinct, but partially overlapping, populations of PNNs. Based on the reactivities of these probes, we categorized PNNs into four groups. We found that two subpopulation of PNNs, one with higher and one with lower WFA-staining are differentially labeled by Cat316 and Cat315, respectively. CS chains recognized by Cat316 were diminished in mice deficient in an enzyme involved in the initiation of CS-biosynthesis. Furthermore, WFA+ and Cat316+ aggrecan were spatially segregated and formed microdomains in a single PNN. Otx2 co-localized with Cat316+ but not with WFA+ aggrecan in PNNs. Our results suggest that the heterogeneity of PNNs around PV cells may affect the functional maturation of these cells.

Sulphate Incorporation in Borosilicate Glasses and Melts: a Kinetic Approach

Energy Technology Data Exchange (ETDEWEB)

Lenoir, M. [CEA, DEN, Laboratoire d' etude et de Developpement de Matrices de Conditionnement, Centre de Marcoule, 30207 Bagnols-sur-Ceze (France); Physique des Mineraux et Magmas, UMR 7047, CNRS- Institut de Physique du Globe de Paris, 7 place Jussieu, 75252 Paris 05 (France); Grandjean, A. [Institut de Chimie Separative de Marcoule, UMR 5257, Laboratoire des Nanomateriaux Autoreparants, Marcoule, 30207 Bagnols-sur-Ceze (France); Dussossoy, J.L. [CEA, DEN, Laboratoire d' etude et de Developpement de Matrices de Conditionnement, Centre de Marcoule, 30207 Bagnols-sur-Ceze (France); Neuville, D.R. [Physique des Mineraux et Magmas, UMR 7047, CNRS- Institut de Physique du Globe de Paris, 7 place Jussieu, 75252 Paris 05 (France)

2008-07-01

The kinetics of sulphate departure in a sodium borosilicate melt were studied using in situ Raman spectroscopy. This technique allows the quantification of the amount of sulphate dissolved in a borosilicate glass as a function of heating time by comparison with measurements obtained by microprobe wavelength dispersive spectrometry. To quantify the sulphate content obtained with Raman spectroscopy, the integrated intensity of the sulphate band at 990 cm{sup -1} was scaled to the sum of the integrated bands between 800 and 1200 cm{sup -1}, bands that are assigned to Qn silica units on the basis of previous literature. Calibration curves were then determined for two different samples. An evaluation of the kinetics of departure of sulphate could thus be made as a function of the viscosity of the borosilicate glass, showing that the kinetics were controlled by the diffusion of sulphate and its volatilization from the melt. This experimental method allows in situ measurements of sulphate content at high temperature which cannot be obtained by any other simple technique. (authors)

The effect of desulfation of chondroitin sulfate on interactions with positively charged growth factors and upregulation of cartilaginous markers in encapsulated MSCs.

Science.gov (United States)

Lim, Jeremy J; Temenoff, Johnna S

2013-07-01

Sulfated glycosaminoglycans (GAGs) are known to interact electrostatically with positively charged growth factors to modulate signaling. Therefore, regulating the degree of sulfation of GAGs may be a promising approach to tailor biomaterial carriers for controlled growth factor delivery and release. For this study, chondroitin sulfate (CS) was first desulfated to form chondroitin, and resulting crosslinked CS and chondroitin hydrogels were examined in vitro for release of positively charged model protein (histone) and for their effect on cartilaginous differentiation of encapsulated human mesenchymal stem cells (MSCs). Desulfation significantly increased the release of histone from chondroitin hydrogels (30.6 ± 2.3 μg released over 8 days, compared to natively sulfated CS with 20.2 ± 0.8 μg), suggesting that sulfation alone plays a significant role in modulating protein interactions with GAG hydrogels. MSCs in chondroitin hydrogels significantly upregulated gene expression of collagen II and aggrecan by day 21 in chondrogenic medium (115 ± 100 and 23.1 ± 7.9 fold upregulation of collagen II and aggrecan, respectively), compared to CS hydrogels and PEG-based swelling controls, indicating that desulfation may actually enhance the response of MSCs to soluble chondrogenic cues, such as TGF-β1. Thus, desulfated chondroitin materials present a promising biomaterial tool to further investigate electrostatic GAG/growth factor interactions, especially for repair of cartilaginous tissues. Copyright © 2013 Elsevier Ltd. All rights reserved.

Gastrointestinal tract wall visualization and distention during abdominal and pelvic multidetector CT with a neutral barium sulphate suspension: comparison with positive barium sulphate suspension and with water.

Science.gov (United States)

Oliva, M R; Erturk, S M; Ichikawa, T; Rocha, T; Ros, P R; Silverman, S G; Mortele, K J

2012-01-01

When examining patients with contrast-enhanced multidetector-row CT, we determined if the stomach and small bowel were visualized and distended better with a neutral barium sulphate suspension than with positive barium sulphate suspension or water. After obtaining approval from our institutional review board, 156 patients (women: 84; mean age: 54 yrs) with no history of gastrointestinal tract disease were randomized prospectively to receive orally either 900 ml of neutral (0.1% w/v) barium sulphate suspension (n = 53), 900 ml of positive (2.1% w/v) barium sulphate suspension (n = 53), or 900 ml of water (n = 50), prior to undergoing contrast-enhanced abdominal and pelvic multidetector-row CT. Two independent radiologists evaluated the stomach, and small bowel, for luminal distension and wall visualization, using a five point scale. Results were compared using Kruskal-Wallis and Mann-Whitney U tests. The walls of the stomach, and small bowel were visualized better in patients who were administered neutral barium sulphate suspension than those who were administered either positive barium sulphate suspension (p barium sulphate suspension, the stomach and small bowel were distended better compared to patients administered water (p barium sulphate suspension (p contrast-enhanced abdominal and pelvic multidetector-row CT, orally administered neutral barium sulphate suspension allows the gastrointestinal tract to be visualized and distended better than either positive barium sulphate suspension, or water.

Manipulating the extracellular matrix: an animal model of the bladder pain syndrome.

Science.gov (United States)

Offiah, Ifeoma; Didangelos, Athanasios; OʼReilly, Barry A; McMahon, Stephen B

2017-01-01

Bladder pain syndrome (BPS) is associated with breakdown of the protective uroepithelial barrier of the urinary bladder allowing urinary constituents access to bladder sensory neurons. Although there are several animal models of cystitis, none specifically relates to BPS. Here, we aimed to create such a model using enzymatic digestion of the barrier proteoglycans (PGs) in the rat. Twenty female Wistar rats were anaesthetized and transurethrally catheterized. Ten animals were treated with 0.25IU of intravesical chondroitinase ABC and heparanase III to digest chondroitin sulphate and heparin sulphate PGs, respectively. Ten animals received saline. Following PG deglycosylation, bladders showed irregular loss of the apical uroplakin and a significant increase in neutrophils, not evident in the control group. Spinal cord sections were also collected for c-fos analysis. A large and significant increase in fos immunoreactivity in the L6/S1 segments in the treatment vs control bladders was observed. Cystometry was performed on 5 treatment and 5 control animals. Analysis revealed a significant increase in micturition reflex excitability postdeglycosylation. On a further group of 10 animals, von Frey mechanical withdrawal thresholds were tested on abdominal skin before and after PG digestions. There was a significant decrease in abdominal mechanical withdrawal threshold postdeglycosylation compared with controls. The results of this animal study suggest that many of the clinical features of BPS are seen after PG digestion from the bladder lumen. This model can be used to further understand mechanisms of pain in patients with BPS and to test new therapeutic strategies.

Effects of agmatine sulphate on facial nerve injuries.

Science.gov (United States)

Surmelioglu, O; Sencar, L; Ozdemir, S; Tarkan, O; Dagkiran, M; Surmelioglu, N; Tuncer, U; Polat, S

2017-03-01

To evaluate the effect of agmatine sulphate on facial nerve regeneration after facial nerve injury using electron and light microscopy. The study was performed on 30 male Wistar albino rats split into: a control group, a sham-treated group, a study control group, an anastomosis group, and an anastomosis plus agmatine sulphate treatment group. The mandibular branch of the facial nerve was dissected, and a piece was removed for histological and electron microscopic examination. Regeneration was better in the anastomosis group than in the study control group. However, the best regeneration findings were seen in the agmatine sulphate treatment group. There was a significant difference between the agmatine group and the others in terms of median axon numbers (p Agmatine sulphate treatment with anastomosis in traumatic facial paralysis may enhance nerve regeneration.

Similarity of recombinant human perlecan domain 1 by alternative expression systems bioactive heterogenous recombinant human perlecan D1

DEFF Research Database (Denmark)

Ellis, April L; Pan, Wensheng; Yang, Guang

2010-01-01

BACKGROUND: Heparan sulfate glycosaminoglycans are diverse components of certain proteoglycans and are known to interact with growth factors as a co-receptor necessary to induce signalling and growth factor activity. In this report we characterize heterogeneously glycosylated recombinant human...... perlecan domain 1 (HSPG2 abbreviated as rhPln.D1) synthesized in either HEK 293 cells or HUVECs by transient gene delivery using either adenoviral or expression plasmid technology. RESULTS: By SDS-PAGE analysis following anion exchange chromatography, the recombinant proteoglycans appeared to possess...... glycosaminoglycan chains ranging, in total, from 6 kDa to >90 kDa per recombinant. Immunoblot analysis of enzyme-digested high Mr rhPln.D1 demonstrated that the rhPln.D1 was synthesized as either a chondroitin sulfate or heparan sulfate proteoglycan, in an approximately 2:1 ratio, with negligible hybrids. Secondary...

A multi-analytical approach to better assess the keratan sulfate contamination in animal origin chondroitin sulfate.

Science.gov (United States)

Restaino, Odile Francesca; Finamore, Rosario; Diana, Paola; Marseglia, Mariacarmela; Vitiello, Mario; Casillo, Angela; Bedini, Emiliano; Parrilli, Michelangelo; Corsaro, Maria Michela; Trifuoggi, Marco; De Rosa, Mario; Schiraldi, Chiara

2017-03-15

Chondroitin sulfate is a glycosaminoglycan widely used as active principle of anti-osteoarthritis drugs and nutraceuticals, manufactured by extraction from animal cartilaginous tissues. During the manufacturing procedures, another glycosaminoglycan, the keratan sulfate, might be contemporarily withdrawn, thus eventually constituting a contaminant difficult to be determined because of its structural similarity. Considering the strict regulatory rules on the pureness of pharmaceutical grade chondrotin sulfate there is an urgent need and interest to determine the residual keratan sulfate with specific, sensitive and reliable methods. To pursue this aim, in this paper, for the first time, we set up a multi-analytical and preparative approach based on: i) a newly developed method by high performance anion-exchange chromatography with pulsed amperometric detection, ii) gas chromatography-mass spectrometry analyses, iii) size exclusion chromatography analyses coupled with triple detector array module and on iv) strong anion exchange chromatography separation. Varied KS percentages, in the range from 0.1 to 19.0% (w/w), were determined in seven pharmacopeia and commercial standards and nine commercial samples of different animal origin and manufacturers. Strong anion exchange chromatography profiles of the samples showed three or four different peaks. These peaks analyzed by high performance anion-exchange with pulsed amperometric detection and size exclusion chromatography with triple detector array, ion chromatography and by mono- or two-dimensional nuclear magnetic resonance revealed a heterogeneous composition of both glycosaminoglycans in terms of sulfation grade and molecular weight. High molecular weight species (>100 KDa) were also present in the samples that counted for chains still partially linked to a proteoglycan core. Copyright © 2016 The Author(s). Published by Elsevier B.V. All rights reserved.

The reducibility of sulphuric acid and sulphate in aqueous solution

International Nuclear Information System (INIS)

Grauer, R.

1991-07-01

In connection with the Swedish project for final storage of spent fuel elements it was necessary to assess whether dissolved sulphate can corrode the copper canister. A simple reaction between copper and sulphate is thermodynamically impossible, but copper can react to give copper sulphide if an additional electron donor such as iron(II) is available. The problem was extended to the more general question of the reducibility of sulphur(VI) in dilute aqueous solution. Chemical reduction of sulphate does not take place in dilute solution at temperatures below 100 o C. In experiments on the reduction of sulphates under hydrothermal conditions a reaction only takes place at temperatures above 275-300 o C. The oxidising action of sulphuric acid on metals becomes perceptible only at acid concentrations over 45-50%. In experiments on the cathodic reduction of 74% sulphuric acid the formation of hydrogen sulphide and elementary sulphur starts, depending on the current density, at 50-130 o C, and polarographic measurements suggest that the reducible species is not the hydrogen sulphate ion but molecular sulphuric acid. The resistance of copper to oxygen-free sulphuric acid up to a concentration of 60% is well-known. Numerous processes in industrial electrochemistry take place in sulphuric acid or sulphate electrolytes. The reversible metal/metal-sulphate electrodes of lead and cadmium are unstable relative to the corresponding metal sulphides. Nevertheless the reversible lead sulphate electrode does not fail from sulphide formation. All these facts confirm that sulphur(VI) in dilute solution is completely inert towards chemical reducing agents and also to cathodic reduction. Thus corrosion of copper by sulphate under final-storage conditions and in the absence of sulphate reducing bacteria can be almost certainly be ruled out. (author) 5 figs., 85 refs

Effectiveness of intra-articular injections of sodium hyaluronate-chondroitin sulfate in knee osteoarthritis: a multicenter prospective study.

Science.gov (United States)

Rivera, Fabrizio; Bertignone, Luca; Grandi, Giancarlo; Camisassa, Roberto; Comaschi, Guido; Trentini, Diego; Zanone, Marco; Teppex, Giuseppe; Vasario, Gabriele; Fortina, Giorgio

2016-03-01

Intra-articular injection of hyaluronic acid is a well-established therapy for the treatment of knee osteoarthritis. The aim of the study was to assess the effectiveness and safety of the use of Arthrum HCS(®) (40 mg hyaluronic acid and 40 mg chondroitin sulfate in 2 mL). This was an open, multicenter, prospective study. Men or women over 40 years of age with documented knee osteoarthritis and WOMAC subscore A (severity of pain) ≥25 were enrolled. They received three weekly intra-articular injections of sodium hyaluronate 2 % and chondroitin sulfate 2 % in combination. WOMAC subscore A was assessed at 1, 3 and 6 months after the last injection. One hundred and twelve patients were included (women, 66 %). The mean (SD) WOMAC subscore A decreased from 52.1 (15.2) at inclusion to 20.5 (19.7) at month 6 (P chondroitin sulfate in reducing pain (77 %), improving mobility (78 %) and reducing the consumption of analgesics (74 %). Only one adverse effect was reported by one patient (knee tumefaction). These results suggest that intra-articular injections of Arthrum HCS(®) (sodium hyaluronate plus chondroitin sulfate) in patients with knee osteoarthritis are efficient and safe. These results should be confirmed in a randomized controlled study. IV.

Protective effect of exogenous chondroitin 4,6-sulfate in the acute degradation of articular cartilage in the rabbit.

Science.gov (United States)

Uebelhart, D; Thonar, E J; Zhang, J; Williams, J M

1998-05-01

The injection of 2.0 mg chymopapain into the adolescent rabbit knee causes severe loss of articular cartilage proteoglycans (PG). Although chondrocytes attempt to restore lost PG, failure to repair ensues. Pure chondroitin 4,6-sulfate (Condrosulf, IBSA Lugano, Switzerland) has been used in clinical studies of human osteoarthritis (OA) as a slow-acting drug for OA (SYSADOA). Using our model of articular cartilage injury, we examined the effects of oral and intramuscular administration of Condrosulf after chymopapain-induced cartilage injury. In this study, animals received an injection of 2.0 mg chymopapain (Chymodiactin, Boots Pharmaceuticals) into the left knee and were sacrificed after 84 days. The contralateral right knee served as a noninjected control. Some animals received oral Condrosulf while others received intramuscular injections of Condrosulf. Serum keratan sulfate (KS) levels were monitored to ensure degradation of the cartilage PG. Those animals not exhibiting at least a 100% increase of serum KS following chymopapain injection were excluded from the study. At sacrifice, cartilage PG contents were markedly reduced in animals receiving an injection of 2.0 mg chymopapain with no further treatment. In contrast, oral administration of Condrosulf beginning 11 days prior to chymopapain injury resulted in significantly higher (P = 0.0036) cartilage PG contents. Intramuscular administration of Condrosulf resulted in higher, but less significantly so (P = 0.0457), cartilage PG contents. These results suggest that daily Condrosulf treatment prior to and continuing after chymopapain injury may have a protective effect on the damaged cartilage, allowing it to continue to re-synthesize matrix PG after the treatment is discontinued.

Anthropogenic influence on the distribution of tropospheric sulphate aerosol

Energy Technology Data Exchange (ETDEWEB)

Langner, J; Rodhe, H; Crutzen, P J; Zimmermann, P [Swedish Meteorological and Hydrological Institute, Norrkoeping (Sweden)

1992-10-22

Human activities have increased global emissions of sulphur gases by about a factor of three during the past century, leading to increased sulphate aerosol concentrations, mainly in the Northern Hemisphere. Sulphate aerosols can affect the climate directly, by increasing the backscattering of solar radiation in cloud-free air, and indirectly, by providing additional cloud condensation nuclei. Here a global transport-chemistry model is used to estimate the changes in the distribution of tropospheric sulphate aerosol and deposition of non-seasalt sulphur that have occurred since pre-industrial times. The increase in sulphate aerosol concentration is small over the Southern Hemisphere oceans, but reaches a factor of 100 over northern Europe in winter. Calculations indicate, however, that at most 6% of the anthropogenic sulphur emissions is available for the formation of new aerosol particles. This is because about one-half of the sulphur dioxide is deposited on the Earth's surface, and most of the remainder is oxidized in cloud droplets so that the sulphate becomes associated with pre-existing particles. Even so, the rate of formation of new sulphate particles may have doubled since pre-industrial times. 18 refs., 3 figs.

Long-term experience with sodium chondroitin sulfate in patients with painful bladder syndrome.

Science.gov (United States)

Tornero, J I; Olarte, H; Escudero, F; Gómez, G

2013-09-01

To assess the response of patients diagnosed with painful bladder syndrome to treatment with instillations of sodium chondroitin sulfate. We present a series of cases of patients with painful bladder syndrome who followed a bladder instillation protocol with sodium chondroitin sulfate, according to our centre's regimen. The response to treatment was assessed with respect to pain, according to the Downie scale; urinary frequency, according to the voiding diary; and subjective improvement, according to the Patient Global Impression of Improvement (PGI-I) scale. A total of 28 patients with a median age of 59 years (range 22-90) followed this protocol. From the medical histories, 19.4% had suffered an infection of the urinary tract, 3.8% had suffered urinary tuberculosis, 7.6% received pelvic radiation therapy and 26.9% had taken anticholinergic drugs for overactive bladder syndrome. We evaluated the response to treatment at 0, 3, 6 and 12 months and found that at the end of treatment 72.3% of the patients had improved bladder pain and 75% were significantly better. Treatment with sodium chondroitin sulfate through endovesical instillation in painful bladder syndrome improves pain, voiding frequency and quality of life in the long term. Copyright © 2013 AEU. Published by Elsevier Espana. All rights reserved.

Proteoglycans in health and disease: the multiple roles of syndecan shedding

DEFF Research Database (Denmark)

Manon-Jensen, Tina; Itoh, Yoshifumi; Couchman, John R

2010-01-01

Proteolytic processes in the extracellular matrix are a major influence on cell adhesion, migration, survival, differentiation and proliferation. The syndecan cell-surface proteoglycans are important mediators of cell spreading on extracellular matrix and respond to growth factors and other...

sup(113m)indium-iron chondroitin sulfate colloid for quantitative assessment of the marrow RE function

Energy Technology Data Exchange (ETDEWEB)

Okuyama, S; Ito, Y; Takahashi, K; Sato, T; Matsuzawa, T [Tohoku Univ., Sendai (Japan). Research Inst. for Tuberculosis, Leprosy and Cancer

1975-07-01

sup(113m)In-iron chondroitin sulfate colloid shows a large accumulation in the bone marrow and is suitable for bone marrow imaging. Quantitative assessment of the marrow reticuloendotherial function was performed using this compound. When an appropriate amount of iron carrier was added for adjustment, the rate of accumulation of hyperfunction in the marrow reticuloendotherial system (RES) induced by acute loss of blood increased. Marrow RES hypofunction was efficiently exhibited regardless of the presence or absence of iron carrier. Deposition of sup(113m)In-iron chondroitin sulfate in the spleen increased remarkably in the presence of carrier In. sup(113m)In-iron chondroitin sulfate colloid appears to be suitable for the measurement of the conditions of marrow RES functions. If short half-life nuclide radio-colloids of the present type are clinically applied, it is possible not only to elaborately observe the bone marrow by scintigraphy but also to gradually decrease the absorbed dose of irradiation.

sup(113m)indium-iron chondroitin sulfate colloid for quantitative assessment of the marrow RE function

International Nuclear Information System (INIS)

Okuyama, Shinichi; Ito, Yasuhiko; Takahashi, Kunibumi; Sato, Tachio; Matsuzawa, Taiju

1975-01-01

sup(113m)In-iron chondroitin sulfate colloid shows a large accumulation in the bone marrow and is suitable for bone marrow imaging. Quantitative assessment of the marrow reticuloendotherial function was performed using this compound. When an appropriate amount of iron carrier was added for adjustment, the rate of accumulation of hyperfunction in the marrow reticuloendotherial system (RES) induced by acute loss of blood increased. Marrow RES hypofunction was efficiently exhibited regardless of the presence or absence of iron carrier. Deposition of sup(113m)In-iron chondroitin sulfate in the spleen increased remarkably in the presence of carrier In. sup(113m)In-iron chondroitin sulfate colloid appears to be suitable for the measurement of the conditions of marrow RES functions. If short half-life nuclide radio-colloids of the present type are clinically applied, it is possible not only to elaborately observe the bone marrow by scintigraphy but also to gradually decrease the absorbed dose of irradiation. (Mukohata, S.)

Vascular wall proteoglycan synthesis and structure as a target for the prevention of atherosclerosis

Directory of Open Access Journals (Sweden)

Peter J Little

2007-03-01

Full Text Available Peter J Little1, 2, 3, Mandy L. Ballinger1, Narin Osman1,31Cell Biology of Diabetes Laboratory, Baker Heart Research Institute, Melbourne, Australia; Monash University, Departments of 2Medicine and 3Immunology, Central and Eastern Clinical School, Alfred Hospital, Melbourne, AustraliaAbstract: Atherosclerosis is the underlying pathology of most cardiovascular disease and it represents the major cause of premature death in modern societies. Current therapies target risk factors being hypertension, hypercholesterolemia, hypertriglyceridemia and hyperglycemia when diabetes is present however the maximum efficacy of these strategies is often 30% or less. Areas of vascular biology that may lead to the development of a complementary vascular wall directed therapy are: inflammation, oxidation, endothelial dysfunction, diabetes-specific factors —hyperglycemia and advanced glycation endproducts and lipid retention by vascular matrix specifically proteoglycans. The major structural features of proteoglycans that determine low-density lipoprotein (LDL binding are the length and sulfation pattern on the glycosaminoglycan (GAG chains. Emerging data discussed in this review indicates that these structural properties are subject to considerable regulation by vasoactive substances possibly using novel signaling pathways. For example, GAG elongation stimulated by platelet-derived growth factor is not blocked by the receptor tyrosine kinase antagonist, genistein suggesting that there may be a previously unknown signaling pathway involved in this response. Thus, modifying proteoglycan synthesis and structure may represent a prime target to prevent LDL binding and entrapment in the vessel wall and thus prevent the development and progression of atherosclerosis.Keywords: proteoglycans, signaling, lipoproteins, atherosclerosis

Controlling Barium Sulphate Scale Deposition Problems in an unbleached Kraft Paper Mill

CSIR Research Space (South Africa)

Sithole, Bruce

2015-06-01

Full Text Available Troubleshooting of scale deposits and defects in paper samples showed that the problem was caused by barium sulphate and calcium sulphate scales. However, it was ascertained that barium sulphate was more of a concern than calcium sulphate...

Treatment with magnesium sulphate in pre-term birth

DEFF Research Database (Denmark)

Wolf, Hans; Hegaard, H K; Greisen, G

2012-01-01

Premature birth increases a child's risk of cerebral palsy and death. The aim of this work is to investigate the association between treatment with magnesium sulphate during premature deliveries and infants' cerebral palsy and mortality through a meta-analysis of observational studies....... A comprehensive search of the Cochrane Library, EMBASE and the PubMed database from their inceptions to 1 October, 2010 using the keywords 'magnesium sulphate, children/infant/pre-term/premature and cerebral palsy/mortality/morbidity/adverse effects/outcome' identified 11 reports of observational studies. Two...... authors working independently extracted the data. A meta-analysis of the data found an association between magnesium sulphate treatment and a significantly reduced risk of mortality (RR 0.73; 95% CI 0.61-0.89) and cerebral palsy (OR 0.64; 95% CI 0.47-0.89). Antenatal treatment with magnesium sulphate...

Proteoglycans in Leiomyoma and Normal Myometrium

Science.gov (United States)

Barker, Nichole M.; Carrino, David A.; Caplan, Arnold I.; Hurd, William W.; Liu, James H.; Tan, Huiqing; Mesiano, Sam

2015-01-01

Uterine leiomyoma are a common benign pelvic tumors composed of modified smooth muscle cells and a large amount of extracellular matrix (ECM). The proteoglycan composition of the leiomyoma ECM is thought to affect pathophysiology of the disease. To test this hypothesis, we examined the abundance (by immunoblotting) and expression (by quantitative real-time polymerase chain reaction) of the proteoglycans biglycan, decorin, and versican in leiomyoma and normal myometrium and determined whether expression is affected by steroid hormones and menstrual phase. Leiomyoma and normal myometrium were collected from women (n = 17) undergoing hysterectomy or myomectomy. In vitro studies were performed on immortalized leiomyoma (UtLM) and normal myometrial (hTERT-HM) cells with and without exposure to estradiol and progesterone. In leiomyoma tissue, abundance of decorin messenger RNA (mRNA) and protein were 2.6-fold and 1.4-fold lower, respectively, compared with normal myometrium. Abundance of versican mRNA was not different between matched samples, whereas versican protein was increased 1.8-fold in leiomyoma compared with myometrium. Decorin mRNA was 2.4-fold lower in secretory phase leiomyoma compared with proliferative phase tissue. In UtLM cells, progesterone decreased the abundance of decorin mRNA by 1.3-fold. Lower decorin expression in leiomyoma compared with myometrium may contribute to disease growth and progression. As decorin inhibits the activity of specific growth factors, its reduced level in the leiomyoma cell microenvironment may promote cell proliferation and ECM deposition. Our data suggest that decorin expression in leiomyoma is inhibited by progesterone, which may be a mechanism by which the ovarian steroids affect leiomyoma growth and disease progression. PMID:26423601

Sulphate in Liquid Nuclear Waste: from Production to Containment

Energy Technology Data Exchange (ETDEWEB)

Lenoir, M.; Grandjean, A.; Ledieu, A.; Dussossoy, J.L.; Cau Dit Coumes, C.; Barre, Y.; Tronche, E. [CEA Marcoule, DEN/DTCD/SECM/LDMC, Batiment 208 BP17171, Bagnols sur Ceze, 30207 (France)

2009-06-15

Nuclear industry produces a wide range of low and intermediate level liquid radioactive wastes which can include different radionuclides such as {sup 90}Sr. In La Hague reprocessing plant and in the nuclear research centers of CEA (Commissariat a l'Energie Atomique), the coprecipitation of strontium with barium sulphate is the technique used to treat selectively these contaminated streams with the best efficiency. After the decontamination process, low and intermediate level activity wastes incorporating significant quantities of sulphate are obtained. The challenge is to find a matrix easy to form and with a good chemical durability which is able to confine this kind of nuclear waste. The current process used to contain sulphate-rich nuclear wastes is bituminization. However, in order to improve properties of containment matrices and simplify the process, CEA has chosen to supervise researches on other materials such as cements or glasses. Indeed, cements are widely used for the immobilization of a variety of wastes (low and intermediate level wastes) and they may be an alternative matrix to bitumen. Even if Portland cement, which is extensively used in the nuclear industry, presents some disadvantages for the containment of sulphate-rich nuclear wastes (risk of swelling and cracking due to delayed ettringite formation), other cement systems, such as calcium sulfo-aluminate binders, may be valuable candidates. Another matrix to confine sulphate-rich waste could be the glass. One of the advantages of this material is that it could also immobilize sulphate containing high level nuclear waste which is present in some countries. This waste comes from the use of ferrous sulfamate as a reducing agent for the conversion of Pu{sup 4+} to Pu{sup 3+} in the partitioning stage of the actinides during reprocessing. Sulphate solubility in borosilicate glasses has already been studied in CEA at laboratory and pilot scales. At a pilot scale, low level liquid waste has been

Salmon cartilage proteoglycan suppresses mouse experimental colitis through induction of Foxp3+ regulatory T cells

International Nuclear Information System (INIS)

Mitsui, Toshihito; Sashinami, Hiroshi; Sato, Fuyuki; Kijima, Hiroshi; Ishiguro, Yoh; Fukuda, Shinsaku; Yoshihara, Shuichi; Hakamada, Ken-Ichi; Nakane, Akio

2010-01-01

Research highlights: → Salmon proteoglycan suppresses IL-10 -/- cell transfer-induced colitis progression. → Salmon proteoglycan suppresses Th1- and Th17-related factors in colitis mice. → Salmon proteoglycan enhances Foxp3 expression. -- Abstract: Proteoglycans (PGs) are complex glycohydrates which are widely distributed in extracellular matrix (ECM). PGs are involved in the construction of ECM, cell proliferation and differentiation. ECM components are involved in transduction of proinflammatory responses, but it is still unknown whether PGs are involved in inflammatory response. In this study, we investigated the effect of PG extracted from salmon cartilage on the progression of experimental colitis-induced in severe combined immunodeficiency mice by cell transfer from interleukin-10 (IL-10) -/- mice. IL-10 -/- cell-transferred mice showed weight loss, colon shortening and histological appearance of mild colitis. Daily oral administration of PG attenuated the clinical progression of colitis in a dose-dependent manner. Colitis-induced mice showed the elevated expression of IFN-γ, IL-12, TNF-α, IL-21, IL-23p19, IL-6, IL-17A and retinoic acid-related orphan receptor γt (RORγt) in lamina propria mononuclear cells (LPMCs) and oral administration of PG suppressed the expression of these factors. Conversely, expression of Foxp3 that induces CD4 + CD25 + regulatory T cells in LPMCs was enhanced by PG administration. These findings suggested that salmon PG attenuated the progression of colitis due to suppression of inflammatory response by enhancement of regulatory T cell induction.

Post traumatic tetanus and role magnesium sulphate

International Nuclear Information System (INIS)

Sikendr, R.I.; Samad, B.U.; Memon, M.I.

2009-01-01

Tetanus is a life threatening disease. Reported mortality for tetanus is 15-39%. Conventional treatment includes heavy sedation and artificial ventilation. Complications resulting from long term heavy sedation and artificial ventilation contribute to 60% of the total mortality caused by tetanus. In this study magnesium sulphate was used to reduce the need for sedation and artificial ventilation. Objectives of this prospective study were to determine the role of magnesium sulphate in post traumatic tetanus. The study was carried out in surgical Intensive Care at Pakistan Institute of Medical Sciences (PIMS), Islamabad from Jan 2004 to Dec 2007. Forty-four patients presented during this period and 33 patients were included in the study. All patients had tracheostomy done within 48 hours. Every patient was started Magnesium Sulphate therapy for control of spasms after sending baseline investigations. Patients were given ventilatory support when needed. All data was entered in well structured proforma. SPSS-10 was used to analyse data. Thirty-three patients were included in the study and all patients were given magnesium sulphate. Out of these, 45.5% cases were grade 4 tetanus, 73.6% and 63.3% cases did not require artificial ventilation and additional sedation respectively, 51.1% patients remained free of complications of tetanus. Overall mortality was 30.3%. Use of Magnesium Sulphate is safe and reduces the need for sedation and artificial ventilation in high grade tetanus thus contributing to survival benefit in adult post-traumatic tetanus cases. (author)

Heparan sulfate proteoglycans made by different basement-membrane-producing tumors have immunological and structural similarities

DEFF Research Database (Denmark)

Wewer, U M; Albrechtsen, R; Hassell, J R

1985-01-01

in the native basement membrane of surrounding normal murine tissues. Blocking and ELISA assays demonstrated that the antibodies recognized both antigens. Using techniques involving the chemical and enzymatic degradation of 35S-sulfate-labeled glycosaminoglycans, the mouse EHS tumor cells were found to produce...... proteoglycans obtained from these two sources immunoprecipitated the same precursor protein with a molecular mass of 400,000 daltons from 35S-methionine pulse-labeled cells of both tumors. Immunohistochemistry showed the heparan sulfate proteoglycan to be distributed in the extracellular matrix and also...

N-acetylgalactosamine positive perineuronal nets in the saccade-related-part of the cerebellar fastigial nucleus do not maintain saccade gain.

Directory of Open Access Journals (Sweden)

Adrienne Mueller

Full Text Available Perineuronal nets (PNNs accumulate around neurons near the end of developmental critical periods. PNNs are structures of the extracellular matrix which surround synaptic contacts and contain chondroitin sulfate proteoglycans. Previous studies suggest that the chondroitin sulfate chains of PNNs inhibit synaptic plasticity and thereby help end critical periods. PNNs surround a high proportion of neurons in the cerebellar nuclei. These PNNs form during approximately the same time that movements achieve normal accuracy. It is possible that PNNs in the cerebellar nuclei inhibit plasticity to maintain the synaptic organization that produces those accurate movements. We tested whether or not PNNs in a saccade-related part of the cerebellar nuclei maintain accurate saccade size by digesting a part of them in an adult monkey performing a task that changes saccade size (long term saccade adaptation. We use the enzyme Chondroitinase ABC to digest the glycosaminoglycan side chains of proteoglycans present in the majority of PNNs. We show that this manipulation does not result in faster, larger, or more persistent adaptation. Our result indicates that intact perineuronal nets around saccade-related neurons in the cerebellar nuclei are not important for maintaining long-term saccade gain.

Interdependent action of nickel sulphate and X-rays on human lymphoblastoid leukeamic cells

International Nuclear Information System (INIS)

Bensimon, Jacques

1977-01-01

In a first experiment, cells were cultured in media supplemented by nickel sulphate, irradiated in same media and cultured in same media after irradiation. In a second experiment, cells were cultured during 18hrs. in media supplemented by nickel sulphate, and then cells were washed and cultured in normal media where they were irradiated. The nickel sulphate toxicity appears as a creasing function of the nickel sulphate concentration and the nickel sulphate action endurance. The nickel sulphate toxic effect is amplified by X-rays. This amplification is a time function that depends on the X-ray dose, nickel sulphate concentration and period of time from the outset of culture to the irradiation. The nickel sulphate toxic effect appears faster when nickel works after X-rays [fr

Expression of small leucine-rich proteoglycans in rat anterior pituitary gland.

Science.gov (United States)

Horiguchi, Kotaro; Syaidah, Rahimi; Fujiwara, Ken; Tsukada, Takehiro; Ramadhani, Dini; Jindatip, Depicha; Kikuchi, Motoshi; Yashiro, Takashi

2013-01-01

Proteoglycans are components of the extracellular matrix and comprise a specific core protein substituted with covalently linked glycosaminoglycan chains. Small leucine-rich proteoglycans (SLRPs) are a major family of proteoglycans and have key roles as potent effectors in cellular signaling pathways. Research during the last two decades has shown that SLRPs regulate biological functions in many tissues such as skin, tendon, kidney, liver, and heart. However, little is known of the expression of SLRPs, or the characteristics of the cells that produce them, in the anterior pituitary gland. Therefore, we have determined whether SLRPs are present in rat anterior pituitary gland. We have used real-time reverse transcription with the polymerase chain reaction to analyze the expression of SLRP genes and have identified the cells that produce SLRPs by using in situ hybridization with a digoxigenin-labeled cRNA probe. We have clearly detected the mRNA expression of SLRP genes, and cells expressing decorin, biglycan, fibromodulin, lumican, proline/arginine-rich end leucine-rich repeat protein (PRELP), and osteoglycin are located in the anterior pituitary gland. We have also investigated the possible double-staining of SLRP mRNA and pituitary hormones, S100 protein (a marker of folliculostellate cells), desmin (a marker of capillary pericytes), and isolectin B4 (a marker of endothelial cells). Decorin, biglycan, fibromodulin, lumican, PRELP, and osteoglycin mRNA have been identified in S100-protein-positive and desmin-positive cells. Thus, we conclude that folliculostellate cells and pericytes produce SLRPs in rat anterior pituitary gland.

Safety assessment of non-animal chondroitin sulfate sodium: Subchronic study in rats, genotoxicity tests and human bioavailability.

Science.gov (United States)

Miraglia, Niccolò; Bianchi, Davide; Trentin, Antonella; Volpi, Nicola; Soni, Madhu G

2016-07-01

Chondroitin sulfate, an amino sugar polymer made of glucuronic acid and N-acetyl-galactosamine, is used in dietary supplements to promote joint health. Commonly used chondroitin sulfate is of animal origin and can pose potential safety problems including bovine spongiform encephalopathy (BSE). The objective of the present study was to investigate potential adverse effects, if any, of microbial derived chondroitin sulfate sodium (CSS) in subchronic toxicity, genotoxicity and bioavailability studies. In the toxicity study, Sprague Dawley rats (10/sex/group) were gavaged with CSS at dose levels of 0, 250, 500 and 1000 mg/kg body weight (bw)/day for 90-days. No mortality or significant changes in clinical signs, body weights, body weight gain or feed consumption were noted. Similarly, no toxicologically relevant treatment-related changes in hematological, clinical chemistry, urinalysis and organ weights were noted. Macroscopic and microscopic examinations did not reveal treatment-related abnormalities. In vitro mutagenic and clastogenic potentials as evaluated by Ames assay, chromosomal aberration test and micronucleus assay did not reveal genotoxicity of CSS. In pharmacokinetic study in human, CSS showed higher absorption as compared to chondroitin sulfate of animal origin. The results of subchronic toxicity study supports the no-observed-adverse-effect level (NOAEL) for CSS as 1000 mg/kg bw/day, the highest dose tested. Copyright © 2016 Elsevier Ltd. All rights reserved.

Epitope mapping and topographic analysis of VAR2CSA DBL3X involved in P-falciparum placental sequestration

DEFF Research Database (Denmark)

Dahlback, Madeleine; Rask, Thomas Salhøj; Andersen, Pernille

2006-01-01

Pregnancy-associated malaria is a major health problem, which mainly affects primigravidae living in malaria endemic areas. The syndrome is precipitated by accumulation of infected erythrocytes in placental tissue through an interaction between chondroitin sulphate A on syncytiotrophoblasts and a...

Deletion of the basement membrane heparan sulfate proteoglycan type XVIII collagen causes hypertriglyceridemia in mice and humans.

Directory of Open Access Journals (Sweden)

Joseph R Bishop

2010-11-01

Full Text Available Lipoprotein lipase (Lpl acts on triglyceride-rich lipoproteins in the peripheral circulation, liberating free fatty acids for energy metabolism or storage. This essential enzyme is synthesized in parenchymal cells of adipose tissue, heart, and skeletal muscle and migrates to the luminal side of the vascular endothelium where it acts upon circulating lipoproteins. Prior studies suggested that Lpl is immobilized by way of heparan sulfate proteoglycans on the endothelium, but genetically altering endothelial cell heparan sulfate had no effect on Lpl localization or lipolysis. The objective of this study was to determine if extracellular matrix proteoglycans affect Lpl distribution and triglyceride metabolism.We examined mutant mice defective in collagen XVIII (Col18, a heparan sulfate proteoglycan present in vascular basement membranes. Loss of Col18 reduces plasma levels of Lpl enzyme and activity, which results in mild fasting hypertriglyceridemia and diet-induced hyperchylomicronemia. Humans with Knobloch Syndrome caused by a null mutation in the vascular form of Col18 also present lower than normal plasma Lpl mass and activity and exhibit fasting hypertriglyceridemia.This is the first report demonstrating that Lpl presentation on the lumenal side of the endothelium depends on a basement membrane proteoglycan and demonstrates a previously unrecognized phenotype in patients lacking Col18.

Effects of a synthetic bioactive peptide on neurite growth and nerve growth factor release in chondroitin sulfate hydrogels

OpenAIRE

Conovaloff, Aaron W.; Beier, Brooke L.; Irazoqui, Pedro P.; Panitch, Alyssa

2011-01-01

Previous work has revealed robust dorsal root ganglia neurite growth in hydrogels of chondroitin sulfate. In the current work, it was determined whether addition of a synthetic bioactive peptide could augment neurite growth in these matrices via enhanced binding and sequestering of growth factors. Fluorescence recovery after photobleaching studies revealed that addition of peptide slowed nerve growth factor diffusivity in chondroitin sulfate gels, but not in control gels of hyaluronic acid. F...

Mesophilic and thermophilic anaerobic digestion of sulphate-containing wastewaters.

Science.gov (United States)

Colleran, E; Pender, S

2002-01-01

The effect of sulphate at an influent chemical oxygen demand (COD):sulphate ratio of 4 on the operational performance of anaerobic hybrid reactors treating molasses wastewater was investigated under mesophilic and thermophilic conditions in a long-term laboratory-scale study over a 1,081 day period. The presence of sulphate reduced the COD removal efficiency under both mesophilic and thermophilic conditions. At 55 degrees C, effluent acetate levels were consistently greater than 4000 mg l(-1) indicating that thermophilic acetate-utilising methane-producing bacteria (MPB) or sulphate-reducing bacteria (SRB) had not developed in the reactor under the conditions applied. At 37 degrees C, acetate was exclusively utilised by acetoclastic methanogens, whereas H2-utilising SRB predominated over H2-utilising MPB in the competition for hydrogen. By contrast, hydrogenotrophic MPB were shown to outcompete H2-utilising SRB during long-term thermophilic operation. 16SrDNA analysis of the seed sludge and reactor biomass on conclusion of the 37 degrees C and 55 degrees C trials illustrated that the dominant methanogen present on conclusion of the thermophilic trial in the absence of influent sulphate was related to Methanocorpusculum parvuum, and was capable of growth on both acetate and hydrogen. By contrast, an organism closely related to Methanobacterium thermoautotrophicum was the dominant methanogen present in the sulphate-fed reactor on completion of the thermophilic trial.

3D chitosan-gelatin-chondroitin porous scaffold improves osteogenic differentiation of mesenchymal stem cells

Energy Technology Data Exchange (ETDEWEB)

Machado, C B [Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais (Brazil); Ventura, J M G [Department of Ceramics and Glass Engineering, University of Aveiro (Portugal); Lemos, A F [Department of Ceramics and Glass Engineering, University of Aveiro (Portugal); Ferreira, J M F [Department of Ceramics and Glass Engineering, University of Aveiro (Portugal); Leite, M F [Department of Physiology and Biophysics, Institute of Biological Sciences, Federal University of Minas Gerais (Brazil); Goes, A M [Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais (Brazil)

2007-06-01

A porous 3D scaffold was developed to support and enhance the differentiation process of mesenchymal stem cells (MSC) into osteoblasts in vitro. The 3D scaffold was made with chitosan, gelatin and chondroitin and it was crosslinked by EDAC. The scaffold physicochemical properties were evaluated. SEM revealed the high porosity and interconnection of pores in the scaffold; rheological measurements show that the scaffold exhibits a characteristic behavior of strong gels. The elastic modulus found in compressive tests of the crosslinked scaffold was about 50 times higher than the non-crosslinked one. After 21 days, the 3D matrix submitted to hydrolytic degradation loses above 40% of its weight. MSC were collected from rat bone marrow and seeded in chitosan-gelatin-chondroitin 3D scaffolds and in 2D culture plates as well. MSC were differentiated into osteoblasts for 21 days. Cell proliferation and alkaline phosphatase activity were followed weekly during the osteogenic process. The osteogenic differentiation of MSC was improved in 3D culture as shown by MTT assay and alkaline phosphatase activity. On the 21st day, bone markers, osteopontin and osteocalcin, were detected by the PCR analysis. This study shows that the chitosan-gelatin-chondroitin 3D structure provides a good environment for the osteogenic process and enhances cellular proliferation.

3D chitosan-gelatin-chondroitin porous scaffold improves osteogenic differentiation of mesenchymal stem cells.

Science.gov (United States)

Machado, C B; Ventura, J M G; Lemos, A F; Ferreira, J M F; Leite, M F; Goes, A M

2007-06-01

A porous 3D scaffold was developed to support and enhance the differentiation process of mesenchymal stem cells (MSC) into osteoblasts in vitro. The 3D scaffold was made with chitosan, gelatin and chondroitin and it was crosslinked by EDAC. The scaffold physicochemical properties were evaluated. SEM revealed the high porosity and interconnection of pores in the scaffold; rheological measurements show that the scaffold exhibits a characteristic behavior of strong gels. The elastic modulus found in compressive tests of the crosslinked scaffold was about 50 times higher than the non-crosslinked one. After 21 days, the 3D matrix submitted to hydrolytic degradation loses above 40% of its weight. MSC were collected from rat bone marrow and seeded in chitosan-gelatin-chondroitin 3D scaffolds and in 2D culture plates as well. MSC were differentiated into osteoblasts for 21 days. Cell proliferation and alkaline phosphatase activity were followed weekly during the osteogenic process. The osteogenic differentiation of MSC was improved in 3D culture as shown by MTT assay and alkaline phosphatase activity. On the 21st day, bone markers, osteopontin and osteocalcin, were detected by the PCR analysis. This study shows that the chitosan-gelatin-chondroitin 3D structure provides a good environment for the osteogenic process and enhances cellular proliferation.

3D chitosan-gelatin-chondroitin porous scaffold improves osteogenic differentiation of mesenchymal stem cells

International Nuclear Information System (INIS)

Machado, C B; Ventura, J M G; Lemos, A F; Ferreira, J M F; Leite, M F; Goes, A M

2007-01-01

A porous 3D scaffold was developed to support and enhance the differentiation process of mesenchymal stem cells (MSC) into osteoblasts in vitro. The 3D scaffold was made with chitosan, gelatin and chondroitin and it was crosslinked by EDAC. The scaffold physicochemical properties were evaluated. SEM revealed the high porosity and interconnection of pores in the scaffold; rheological measurements show that the scaffold exhibits a characteristic behavior of strong gels. The elastic modulus found in compressive tests of the crosslinked scaffold was about 50 times higher than the non-crosslinked one. After 21 days, the 3D matrix submitted to hydrolytic degradation loses above 40% of its weight. MSC were collected from rat bone marrow and seeded in chitosan-gelatin-chondroitin 3D scaffolds and in 2D culture plates as well. MSC were differentiated into osteoblasts for 21 days. Cell proliferation and alkaline phosphatase activity were followed weekly during the osteogenic process. The osteogenic differentiation of MSC was improved in 3D culture as shown by MTT assay and alkaline phosphatase activity. On the 21st day, bone markers, osteopontin and osteocalcin, were detected by the PCR analysis. This study shows that the chitosan-gelatin-chondroitin 3D structure provides a good environment for the osteogenic process and enhances cellular proliferation

Are high rates of sulphate reduction associated with anaerobic oxidation of methane

Energy Technology Data Exchange (ETDEWEB)

Devol, A H; Ahmed, S I

1981-01-01

Classical models of sulphur diagenesis in marine sediments are based on the assumption that the rate of sulphate reduction is first order with respect to oxidizable particulate organic carbon (POC). This assumption requires that oxidizable POC, sulphate concentration and the sulphate reduction rate be highest at the top of the sulphate reduction zone and decrease exponentially with increasing sediment depth. However, to explain recent observations of concave upwards methane distributions, the anaerobic consumption of methane has been proposed. Furthermore, it has been proposed that this consumption takes place near the bottom of the sulphate reducing zone where sulphate concentrations are low. Thus, if sulphate reducing bacteria are associated with the anaerobic oxidation of methane, a peak in sulphate reduction rate might be expected in this deep consumption zone. The importance of the process in sedimentary sulphur diagenesis is indicated by calculations estimating that 30 to 75% of the downward sulphate flux at depth may be consumed by methane oxidation within this zone. We present here profiles of sulphate reduction rate in anoxic sediments that show distinct local maxima at the depth where the anaerobic oxidation of methane would be expected. Our measurements were made during July and August 1978 in Saanich Inlet, an anoxic fjord located on the south-east of Vancouver Island, British Columbia. The inlet has a shallow sill (approx 70 m) which restricts circulation of the deeper water (maximum depth 225 m) inside the basin to the extent that for about 8 months of the year the bottom waters contain hydrogen sulphide, the inlet is an ideal location for studying sedimentary sulphate reduction because reactions with oxygen and the effects of burrowing organisms can be neglected.

Removal of radium from aqueous sulphate solutions

International Nuclear Information System (INIS)

Weir, D.R.; Masters, J.T.; Neven, M.

1983-01-01

Radium is often present in ores and an aqueous solution associated with the ore may consequently contain dissolved radium. It is frequently necessary to remove radium from such solutions to reduce the total radium content to a prescribed low level before the solution can be returned to the environment. The present invention is based on the discovery that the total radium content can be reduced to a satisfactory level within a reasonable time by adding a soluble barium salt to a radium-containing sulphate solution which also contains dissolved magnesium at a pH not greater than about 0 to precipitate radium as barium radium sulphate, raising the pH to at least 11 to precipitate an insoluble magnesium compound which collects the barium radium sulphate precipitate, and separating substantially all of the precipitates from the solution

Computerised analysis of sulphate action on model concrete piles

Digital Repository Service at National Institute of Oceanography (India)

Deshmukh, A.M.; Balasubramanian, S.; Venugopal, C.

Sulphate action severely affects the durability of marine structures. It is imperative to study precisely the effects of magnesium sulphate on pile foundations in the marine environment. In the present paper an attempt is made to assess and analyse...

Bacterial sulphate reduction and the development of alkalinity. II. Laboratory experiments with soils

Energy Technology Data Exchange (ETDEWEB)

Abd-El-Malek, Y; Rizk, S G

1963-01-01

In waterlogged soils sulphate reduction was characterized by increases in counts of sulphate reducers, and increase in titratable alkalinity, insoluble carbonate, and soluble sulphides, and a decrease in soluble Ca/sup 2 +/ + Mg/sup 2 +/. Presence of organic matter greatly enhanced sulphate reduction. A linear relationship between the amount of sulphate reduced and increases in titratable alkalinity and insoluble carbonate were apparent. The potential effects of sulphate reduction on soil fertility through the development of alkalinity are discussed.

Safety and efficacy of Bolus administration of magnesium sulphate ...

African Journals Online (AJOL)

Safety and efficacy of Bolus administration of magnesium sulphate for preeclampsia. ... On-going research is addressing its administration in terms of dosage, duration and safety. Objective: We evaluated a ... Keywords: safety, efficacy, bolus magnesium sulphate, preeclampsia, University of Benin Teaching Hospital

Kinetics and mechanism of protection of thymine from sulphate ...

Indian Academy of Sciences (India)

Unknown

The rate constant of sulphate radical anion with caffeic acid has been ... transfer of radical sites from base moiety to sugar moiety. Strand ... titrated with a standard ceric ammonium sulphate solution as reported by Kapoor et al.7. At room ...

Bacterial sulphate reduction and the development of alkalinity. I. Experiments with synthetic media

Energy Technology Data Exchange (ETDEWEB)

Abd-El-Malek, Y; Rizk, S G

1963-01-01

In liquid cultures of desulphovibrio desulphuricans the effects of the following on sulphate reduction were determined: The type of cation (ca++ or na+) attached to the sulphate ion; the presence of calcium carbonate alone and with sodium chloride; the concentration of sodium sulphate, sodium lactate and sodium chloride; and the initial pH. The titratable alkalinity formed as a result of sulphate reduction was found to be quantitatively related to the sulphate reduced, and apparently unaffected by strain variation or energy source.

Sulphation reactions of oxidic dust particles in waste heat boiler environment. Literature review

Energy Technology Data Exchange (ETDEWEB)

Ranki, T.

1999-09-01

Sulphation of metal oxides has an important role in many industrial processes. In different applications sulphation reactions have different aims and characteristics. In the flash smelting process sulphation of oxidic flue dust is a spontaneous and inevitable phenomena, which takes place in the waste heat boiler (WHB) when cooling down hot dust laden off-gases from sulphide smelters. Oxidic dust particles (size 0 - 50 {mu}m) react with O{sub 2} and SO{sub 2} or SO{sub 3} in a certain temperature range (500 - 800 deg C). Sulphation reactions are highly exothermic releasing large amount of heat, which affects the gas cooling and thermal performance of the boiler. Thermodynamics and kinetics of the system have to be known to improve the process and WHB operation. The rate of sulphation is affected by the prevailing conditions (temperature, gas composition) and particle size and microstructure (porosity, surface area). Some metal oxides (CuO) can react readily with SO{sub 2} and O{sub 2} and act as self-catalysts, but others (NiO) require the presence of an external catalyst to enhance the SO{sub 3} formation and sulphation to proceed. Some oxides (NiO) sulphate directly, some (CuO) may form first intermediate phases (basic sulphates) depending on the reaction conditions. Thus, the reaction mechanisms are very complex. The aim of this report was to search information about the factors affecting the dust sulphation reactions and suggested reaction mechanisms and kinetics. Many investigators have studied sulphation thermodynamics and reaction kinetics and mechanisms of macroscopical metal oxide pieces, but only few articles have been published about sulphation of microscopical particles, like dust. All the found microscale studies dealt with sulphation reactions of calcium oxide, which is not present in the flash smelting process, but used as an SO{sub 2} absorbent in the combustion processes. However, also these investigations may give some hints about the sulphation

Undersulfation of proteoglycans and proteins alter C6 glioma cells proliferation, adhesion and extracellular matrix organization.

Science.gov (United States)

Mendes de Aguiar, Claudia B N; Garcez, Ricardo Castilho; Alvarez-Silva, Marcio; Trentin, Andréa Gonçalves

2002-11-01

Proteoglycans are considered to be important molecule in cell-microenvironment interactions. They are overexpressed in neoplastic cells modifying their growth and migration in hosts. In this work we verified that undersulfation of proteoglycans and other sulfated molecules, induced by sodium chlorate treatment, inhibited C6 glioma cells proliferation in a dose-dependent way. This effect was restored by the addition of exogenous heparin. We could not detect significant cell mortality in our culture condition. The treatment also impaired in a dose-dependent manner, C6 cell adhesion to extracellular matrix (ECM) proteins (collagen IV, laminin and fibronectin). In addition, sodium chlorate treatment altered C6 glioma cell morphology, from the fibroblast-like to a more rounded one. This effect was accompanied by increased synthesis of fibronectin and alterations in its extracellular network organization. However, we could not observe modifications on laminin organization and synthesis. The results suggest an important connection between sulfation degree with important tumor functions, such as proliferation and adhesion. We suggest that proteoglycans may modulate the glioma microenvironment network during tumor cell progression and invasion.

Influence of cytochalasin D-induced changes in cell shape on proteoglycan synthesis by cultured articular chondrocytes

International Nuclear Information System (INIS)

Newman, P.; Watt, F.M.

1988-01-01

There is growing evidence that cell shape regulates both proliferation and differentiated gene expression in a variety of cell types. The authors have explored the relationship between the morphology of articular chondrocytes in culture and the amount and type of proteoglycan they synthesize, using cytochalasin D to induce reversible cell rounding. When chondrocytes were prevented from spreading or when spread cells were induced to round up, 35 SO 4 incorporation into proteoglycan was stimulated. Incorporation into the cell layer was stimulated more than into the medium. When the cells were allowed to respread by removing cytochalasin D, proteoglycan synthesis returned to control levels. Cytochalasin D-induced stimulation of 35 SO 4 incorporation reflected an increase in core protein synthesis rather than lengthening of glycosaminoglycan chains, because [ 3 H]serine incorporation into core protein was also stimulated. Cytochalasm D-treatment of cells in suspension caused no further stimulation of 35 SO 4 incorporation, suggesting that the observed effects were due to cell rounding rather than exposure to cytochalasin D per se

Comparison of functional assays used in the clinical development of a placental malaria vaccine

DEFF Research Database (Denmark)

Pehrson, Caroline; Heno, Kristine Klysner; Adams, Yvonne

2017-01-01

BACKGROUND: Malaria in pregnancy is associated with significant morbidity in pregnant women and their offspring. Plasmodium falciparum infected erythrocytes (IE) express VAR2CSA that mediates binding to chondroitin sulphate A (CSA) in the placenta. Two VAR2CSA-based vaccines for placental malaria...

Novel adenovirus encoded virus-like particles displaying the placental malaria associated VAR2CSA antigen

DEFF Research Database (Denmark)

Andersson, Anne-Marie C; dos Santos Marques Resende, Mafalda; Salanti, Ali

2017-01-01

The malaria parasite Plasmodium falciparum presents antigens on the infected erythrocyte surface that bind human receptors expressed on the vascular endothelium. The VAR2CSA mediated binding to a distinct chondroitin sulphate A (CSA) is a crucial step in the pathophysiology of placental malaria a...

The effects of three-month oral supplementation with a nutraceutical and exercise on the locomotor pattern of aged horses

NARCIS (Netherlands)

Higler, M H; Brommer, H; L'Ami, J J; de Grauw, J C; Nielen, M; van Weeren, P R; Laverty, S; Barneveld, A; Back, W

REASONS FOR PERFORMING STUDY: Multiple in vitro studies assessing articular tissues have indicated that glucosamine and chondroitin sulphate may possess anti-inflammatory effects, but little is known of their clinical effects in vivo. Many old horses have stiff joints, which is likely to be

Interactions of oversulfated chondroitin sulfate (OSCS) from different sources with unfractionated heparin.

Science.gov (United States)

Gray, Angel; Litinas, Evangelos; Jeske, Walter; Fareed, Jawed; Hoppensteadt, Debra

2012-01-01

In 2008, oversulfated chondroitin sulfate (OSCS) was identified as the main contaminant in recalled heparin. Oversulfated chondroitin sulfate can be prepared from bovine (B), porcine (P), shark (Sh), or skate (S) origin and may produce changes in the antithrombotic, bleeding, and hemodynamic profile of heparins. This study examines the interactions of various OSCSs on heparin in animal models of thrombosis and bleeding, as well as on the anticoagulant and antiprotease effects in in vitro assays. Mixtures of 70% unfractionated heparin (UFH) with 30% OSCS from different sources were tested. In the in vitro activated partial thromboplastin time (aPTT) assay, all contaminant mixtures showed a decrease in clotting times. In addition, a significant increase in bleeding time compared to the control (UFH/saline) was observed. In the thrombosis model, no significant differences were observed. The OSCSs significantly increased anti-Xa activity in ex vivo blood samples. These results indicate that various sources of OSCS affect the hemostatic properties of heparin.

Thermal decomposition of uranyl sulphate hydrate

International Nuclear Information System (INIS)

Sato, T.; Ozawa, F.; Ikoma, S.

1980-01-01

The thermal decomposition of uranyl sulphate hydrate (UO 2 SO 4 .3H 2 O) has been investigated by thermogravimetry, differential thermal analysis, X-ray diffraction and infrared spectrophotometry. As a result, it is concluded that uranyl sulphate hydrate decomposes thermally: UO 2 SO 4 .3H 2 O → UO 2 SO 4 .xH 2 O(2.5 = 2 SO 4 . 2H 2 O → UO 2 SO 4 .H 2 O → UO 2 SO 4 → α-UO 2 SO 4 → β-UO 2 SO 4 → U 3 O 8 . (author)

Thermometric titration of sulphate.

Science.gov (United States)

Williams, M B; Janata, J

1970-06-01

Direct thermometric titration of sulphate with a solution of barium perchlorate is proposed. The stoichiometry of the titration is shown to be critically dependent on the concentration of ethanol in the titration medium. The titration is rapid and suffers from only a few interferences.

Magnesium sulphate for fetal neuroprotection

DEFF Research Database (Denmark)

Bickford, Celeste D; Magee, Laura A; Mitton, Craig

2013-01-01

of cerebral palsy (CP) averted and quality-adjusted life years (QALYs). RESULTS: From a health system and a societal perspective, respectively, a savings of $2,242 and $112,602 is obtained for each QALY gained and a savings of $30,942 and $1,554,198 is obtained for each case of CP averted when magnesium......BACKGROUND: The aim of this study was to assess the cost-effectiveness of administering magnesium sulphate to patients in whom preterm birth at ... sensitivity analyses were used to compare the administration of magnesium sulphate with the alternative of no treatment. Two separate cost perspectives were utilized in this series of analyses: a health system and a societal perspective. In addition, two separate measures of effectiveness were utilized: cases...

Sulphate, more than a nutrient, protects the microalga Chlamydomonas moewusii from cadmium toxicity

Energy Technology Data Exchange (ETDEWEB)

Mera, Roi; Torres, Enrique, E-mail: torres@udc.es; Abalde, Julio

2014-03-01

Highlights: • Sulphate effect on cadmium toxicity in the microalga Chlamydomonas moewusii Gerloff. • Cadmium increases the sulphur requirements in Chlamydomonas moewusii. • Kinetic coefficients for sulphate utilization and cadmium effect on them. • Sulphate and cadmium influence on the biosynthesis of low-molecular mass thiols. • Cadmium toxicity reduction by sulphate due to higher biosynthesis of thiols. - Abstract: Sulphur is an essential macroelement that plays important roles in living organisms. The thiol rich sulphur compounds, such as cysteine, γ-Glu–Cys, glutathione and phytochelatins participate in the tolerance mechanisms against cadmium toxicity. Plants, algae, yeasts and most prokaryotes cover their demand for reduced sulphur by reduction of inorganic sulphate. The aim of this study was to investigate, using a bifactorial experimental design, the effect of different sulphate concentrations in the nutrient solution on cadmium toxicity in the freshwater microalga Chlamydomonas moewusii. Cell growth, kinetic parameters of sulphate utilization and intracellular concentrations of low-molecular mass thiol compounds were determined. A mathematical model to describe the growth of this microalga based on the effects of sulphate and cadmium was obtained. An ANOVA revealed an interaction between them, 16% of the effect sizes was explained by this interaction. A higher amount of sulphate in the culture medium allowed a higher cadmium tolerance due to an increase in the thiol compound biosynthesis. The amount of low-molecular mass thiol compounds, mainly phytochelatins, synthesized by this microalga was significantly dependent on the sulphate and cadmium concentrations; the higher phytochelatin content was obtained in cultures with 4 mg Cd/L and 1 mM sulphate. The maximum EC{sub 50} value (based on nominal cadmium concentration) reached for this microalga was 4.46 ± 0.42 mg Cd/L when the sulphate concentration added to the culture medium was also 1 m

Relationship between sulphate and sulphur dioxide in the air

Energy Technology Data Exchange (ETDEWEB)

Fugas, M; Gentilizza, M

1978-01-01

The relationship between the sulphate in suspended particulates and sulphur dioxide in the air was studied in various urban and industrial areas. The relationship is best described by the equation y = ax/sup b/, where y is the percentage of the sulphate S in the total S (sulphate and sulphur dioxide) and x is the concentration of the total S in the air. The regression coefficients a and b seem to be characteristics of the area. In urban areas studied so far a was between 316 and 378 and b between -0.74 and -0.83. In industrial areas polluted by dust which contains elevated concentrations of metals a was between 91 and 107 and b between -0.35 and -0.49. In the area polluted by cement dust there was practically no correlation between the sulphate S (%) and the total S, but a relatively high correlation between absolute amounts of the sulphate S and the total S. The relations indicate that the limitation of SO/sub 2/ conversion is influenced by aerosol composition. Aerosols containing certain metals may promote the conversion by a catalytic effect while alkaline substances by increasing the pH. Whether this can only happen in the plume or in the air as well remains to be clarified.

Salmon cartilage proteoglycan suppresses mouse experimental colitis through induction of Foxp3{sup +} regulatory T cells

Energy Technology Data Exchange (ETDEWEB)

Mitsui, Toshihito [Department of Microbiology and Immunology, Hirosaki University Graduate School of Medicine, Zaifu-cho 5, Hirosaki, Aomori 036-8562 (Japan); Department of Digestive Surgery, Hirosaki University Graduate School of Medicine, Hirosaki, Aomori 036-8562 (Japan); Sashinami, Hiroshi [Department of Microbiology and Immunology, Hirosaki University Graduate School of Medicine, Zaifu-cho 5, Hirosaki, Aomori 036-8562 (Japan); Sato, Fuyuki; Kijima, Hiroshi [Department of Pathology and Bioscience, Hirosaki University Graduate School of Medicine, Hirosaki, Aomori 036-8562 (Japan); Ishiguro, Yoh; Fukuda, Shinsaku [Department of Digestive Internal Medicine, Hirosaki University Graduate School of Medicine, Hirosaki, Aomori 036-8562 (Japan); Yoshihara, Shuichi [Department of Glycomedicine, Hirosaki University Graduate School of Medicine, Hirosaki, Aomori 036-8562 (Japan); Hakamada, Ken-Ichi [Department of Digestive Surgery, Hirosaki University Graduate School of Medicine, Hirosaki, Aomori 036-8562 (Japan); Nakane, Akio, E-mail: a27k03n0@cc.hirosaki-u.ac.jp [Department of Microbiology and Immunology, Hirosaki University Graduate School of Medicine, Zaifu-cho 5, Hirosaki, Aomori 036-8562 (Japan)

2010-11-12

Research highlights: {yields} Salmon proteoglycan suppresses IL-10{sup -/-} cell transfer-induced colitis progression. {yields} Salmon proteoglycan suppresses Th1- and Th17-related factors in colitis mice. {yields} Salmon proteoglycan enhances Foxp3 expression. -- Abstract: Proteoglycans (PGs) are complex glycohydrates which are widely distributed in extracellular matrix (ECM). PGs are involved in the construction of ECM, cell proliferation and differentiation. ECM components are involved in transduction of proinflammatory responses, but it is still unknown whether PGs are involved in inflammatory response. In this study, we investigated the effect of PG extracted from salmon cartilage on the progression of experimental colitis-induced in severe combined immunodeficiency mice by cell transfer from interleukin-10 (IL-10){sup -/-} mice. IL-10{sup -/-} cell-transferred mice showed weight loss, colon shortening and histological appearance of mild colitis. Daily oral administration of PG attenuated the clinical progression of colitis in a dose-dependent manner. Colitis-induced mice showed the elevated expression of IFN-{gamma}, IL-12, TNF-{alpha}, IL-21, IL-23p19, IL-6, IL-17A and retinoic acid-related orphan receptor {gamma}t (ROR{gamma}t) in lamina propria mononuclear cells (LPMCs) and oral administration of PG suppressed the expression of these factors. Conversely, expression of Foxp3 that induces CD4{sup +}CD25{sup +} regulatory T cells in LPMCs was enhanced by PG administration. These findings suggested that salmon PG attenuated the progression of colitis due to suppression of inflammatory response by enhancement of regulatory T cell induction.

Critical appraisal of the role of glucosamine and chondroitin in the management of osteoarthritis of the knee

Directory of Open Access Journals (Sweden)

Steven J Narvy

2010-02-01

Full Text Available Steven J Narvy1, C Thomas Vangsness Jr21Keck School of Medicine, University of Southern California, Los Angeles, CA, USA; 2Department of Orthopaedic Surgery, Keck School of Medicine, University of Southern California, Los Angeles, CA, USAAbstract: Osteoarthritis (OA is the most common musculoskeletal disease in the United States, with rising prevalence. Medical management of OA involves acetaminophen, nonsteroidal anti-inflammatory drugs, and other analgesics, all of which are of variable efficacy and are associated with significant side effects and toxicities. The purpose of this review is to critically evaluate the efficacy of glucosamine and chondroitin, both as single agents and in combination, for the treatment of knee OA. Also evaluated were the level of evidence and funding support of the included articles. Almost every included trial of glucosamine sulfate, glucosamine hydrochloride, and chondroitin sulfate has found the safety of these compounds to be equal to that of placebo, though their therapeutic efficacy in decreasing knee OA pain and improving joint function is variable. Additionally, there are data to support a role of these agents in reducing radiographic progression of knee OA. Industry involvement, however, remains prominent. Further, more comprehensive study by independent researchers free of industry ties is necessary to identify a subset of patients in whom the use of glucosamine and/or chondroitin would be most beneficial. These agents may be safely tried as an initial therapy in select OA patients prior to initiating therapy with nonsteroidal anti-inflammatory drugs, acetaminophen, and other traditional medications.Keywords: glucosamine sulfate, glucosamine hydrochloride, chondroitin sulfate, knee osteoarthritis, nutritional supplement, nutraceutical

[Glycosaminoglycans in subepithelial opacity after excimer laser keratectomy].

Science.gov (United States)

Nakayasu, K; Gotoh, T; Ishikawa, T; Kanai, A

1996-05-01

We evaluated histochemically the characteristics of glycosaminoglycans and proteoglycans in the corneal subepithelial opacity after excimer laser keratectomy on rabbit corneas. We also performed the same evaluations on the cornea after mechanical keratectomy. Twenty days after the operations, the area immediately subjacent to the epithelium showed strong staining with toluidine blue, alcian blue, and colloidal iron. However, after treatment with chondroitinase ABC or chondroitinase AC, alcian blue staining in this area decreased dramatically. Antilarge proteoglycan antibody also reacted strongly in this area. Histochemical and immunohistochemical examination of the cornea where mechanical keratectomy was done showed basically similar findings with the cornea of excimer laser keratectomy. These results suggest that large-molecula proteoglycans with chondroitine sulfate side chains become localized in the subepithelial area after two different kinds of keratectomies. We presume from histochemical and immunohistochemical observations that the subepithelial opacity observed after excimer laser keratectomy is not a special reaction to excimer laser but simply a corneal scar formed after stromal resection.

Feasibility of lead removal from industrial effluents by sulphate reducing bacteria

International Nuclear Information System (INIS)

Nunez, L. A.; Diez, M.; Rodriguez, F. J.

2009-01-01

Battery manufacturing wastewater contains high concentration of sulphate and lead at low pH values. Biogical treatment by anaerobic reduction of sulphate to sulphite, with organic matter as electron donor, increases alkalinity and allow precipitating lead as sulphide. the type of organic matter, COD/SO 4 ratio and lead concentration over sulphate reduction rate are investigated in this work. (Author)

Differential responses of freshwater wetland soils to sulphate pollution

NARCIS (Netherlands)

Lamers, L.P.M.; Dolle, ten G.E.; Berg, van den S.T.G.; Delft, van S.P.J.; Roelofs, J.G.M.

2001-01-01

Sulphate (SO42-) reduction rates are generally low in freshwater wetlands and are regulated by the scarce availability of the ion. Increased concentrations of this electron acceptor due to sulphur (S) pollution of groundwater and surface water may, however, lead to high sulphate reduction rates now

Incorporation of 35S-sulfate and 3H-glucosamine into heparan and chondroitin sulfates during the cell cycle of B16-F10 cells

International Nuclear Information System (INIS)

Blair, O.C.; Sartorelli, A.C.

1984-01-01

Changes in glycosaminoglycan composition occurring during the cell cycle were determined in B16-F10 cells sorted flow cytometrically with respect to DNA content. Incorporation of 35 S-sulfate into heparan sulfate and chondroitin sulfate of unsorted and G1,S, and G2 +M sorted cells was determined following chondroitinase ABC or nitrous acid treatment; the incorporation into surface material was measured as the difference between the radioactivity of control and trypsin-treated cells. Incorporation of 35 S-sulfate and 3 H-glucosamine into cetyl pyridinium chloride (CPC)-precipitable material was characterized before and after chondroitinase or nitrous acid treatment by Sephadex G50 chromatography. Long-term (48 h) and short-term (1 h) labeling studies demonstrate that (a) the amount of total cellular chondroitin sulfate is greater than that of heparan sulfate, with larger amounts of unsulfated heparan than chondroitin being present; (b) the rate of turnover of heparan sulfate is greater than that of chondroitin sulfate; (c) greatest short-term incorporation of 3H-glucosamine into CPC-precipitable material occurs during S phase; and (d) the rate of turnover of both heparan sulfate and chondroitin sulfate is decreased in S phase relative to G1 and G2 + M

The effect of magnesium on partial sulphate removal from mine water as gypsum.

Science.gov (United States)

Tolonen, Emma-Tuulia; Rämö, Jaakko; Lassi, Ulla

2015-08-15

The aim of this research was to investigate the effect of magnesium on the removal efficiency of sulphate as gypsum from mine water. The precipitation conditions were simulated with MINEQL + software and the simulation results were compared with the results from laboratory jar test experiments. Both the simulation and the laboratory results showed that magnesium in the mine water was maintaining sulphate in a soluble form as magnesium sulphate (MgSO4) at pH 9.6. Thus magnesium was preventing the removal of sulphate as gypsum (CaSO4·2H2O). However, change in the lime precipitation pH from 9.6 to 12.5 resulted in magnesium hydroxide (Mg(OH)2) precipitation and improved sulphate removal. Additionally, magnesium hydroxide could act as seed crystals for gypsum precipitation or co-precipitate sulphate further enhancing the removal of sulphate from mine water. Copyright © 2015 Elsevier Ltd. All rights reserved.

Characterization of Glycan Structures of Chondroitin Sulfate-Glycopeptides Facilitated by Sodium Ion-Pairing and Positive Mode LC-MS/MS

Science.gov (United States)

Nilsson, Jonas; Noborn, Fredrik; Gomez Toledo, Alejandro; Nasir, Waqas; Sihlbom, Carina; Larson, Göran

2017-02-01

Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. We have described a protocol to enrich, hydrolyze by chondroitinase ABC, and characterize chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS. The CS-glycopeptides, originating from the Bikunin proteoglycan of human urine samples, had ΔHexAGalNAcGlcAGalGalXyl- O-Ser hexasaccharide structure and were further substituted with 0-3 sulfate and 0-1 phosphate groups. However, it was not possible to exactly pinpoint sulfate attachment residues, for protonated precursors, due to extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD). To circumvent the well-recognized sulfate instability, we now introduced Na+ ions to form sodiated precursors, which protected sulfate groups from decomposition and facilitated the assignment of sulfate modifications. Sulfate groups were pinpointed to both Gal residues and to the GalNAc of the hexasaccharide structure. The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. We have demonstrated a considerable heterogeneity of the bikunin CS linkage region. The realization of these structural variants should be beneficial in studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins. Also, the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis will likely become useful for additional classes of sulfated glycopeptides.

Microwave assisted synthesis of nano sized sulphate doped hydroxyapatite

International Nuclear Information System (INIS)

Alshemary, Ammar Z.; Goh, Yi-Fan; Akram, Muhammad; Razali, Ili Rabihah; Abdul Kadir, Mohammed Rafiq; Hussain, Rafaqat

2013-01-01

Highlights: ► Phase pure nano-sized sulphur doped hydroxyapatite has been synthesized. ► TEM analysis confirmed formation of needle shaped structure. ► Lattice parameters and cell volume increased with increase in sulphate doping. ► Crystallite size decreased as sulphate content inside the structure increased. ► Degree of crystallinity decreased with increase in sulphate substitution. - Abstract: Inorganic sulphate is required by all mammalian cells to function properly, it is the fourth most abundant anion in the human plasma. Sulphate ions are the major source of sulphur which is considered an important element for sustenance of life as it is present in the essential amino and is required by cells to function properly. In this study we have successfully substituted sulphate ions (SO 4 2− ) into hydroxyapatite (Ca 10 (PO 4 ) 6−x (SO 4 ) x (OH) 2−x ) lattice via ion exchange process with phosphate group. Concentration of SO 4 2− ions was varied between X = 0.05–0.5, using (Ca (NO 3 ) 2 ·4H 2 O), ((NH 4 ) 2 HPO 4 ) and (Na 2 SO 4 ) as starting materials. X-ray diffraction (XRD), Fourier transform IR spectroscopy (FTIR), showed that the substitution of SO 4 2− ions into the lattice resulted in peak broadening and reduced peak height due to the amorphous nature and reduced crystallinity of the resulting HA powder. Transmission electron microscopy (TEM) and field emission electron microscopy (FESEM) analysis confirmed the formation of needle shaped particles of 41 nm size with homogenous and uniform distribution of element within the HA structure

Sulphate removal from uraniferous liquors

International Nuclear Information System (INIS)

Berger, B.

1983-01-01

A process for the recovery of uranium from liquor resulting from the attack of sulphur containing uraniferous ores by an alkaline solution of sodium carbonate and/or sodium bicarbonate is claimed. An ion exchange resin is used to separate the uranium from the solution of sodium carbonate and/or bicarbonate and sodium sulphate. The ion exchange resin is then eluted with a solution of ammonium carbonate and/or bicarbonate to provide an eluate containing ammonium uranyl tricarbonate, ammonium carbonate and/or bicarbonate and ammonium sulphate. The eluate is heated to boiling to convert the ammonium uranyl tricarbonate to ammonium uranate and/or diuranate. Ammonia, carbon dioxide and water vapor are released. The precipitated ammonium uranate and/or diuranate is separated from the remaining liquor and calcined to give uranium trioxide

Serum and urinary oestrone sulphate in pregnancy and delivery measured by a direct radioimmunoassay

International Nuclear Information System (INIS)

Honjo, Hideo; Kitawaki, Jo; Itoh, Masafumi; Yasuda, Jinsuke; Yamamoto, Takara; Yamamoto, Takao; Okada, Hiroji; Ohkubo, Tadashi; Nambara, Toshio

1986-01-01

Serum and urinary levels of oestrone sulphate in pregnancy and delivery were measured by a direct radioimmunoassay without hydrolysis. the serum and urinary oestrone sulphate increased as pregnancy progressed. the mean level of serum oestrone sulphate increased to the highest peak of 494 pmol/ml at the 35th gestational week and then decreased. The mean level of urinary oestrone sulphate increased to the highest peak of 1.28 μmol/l at the 34th gestational week and the decreased. At vaginal deliveries, the mean level of maternal peripheral serum oestrone sulphate increased hourly at as high a level as 979 pmol/ml. The mean serum level of oestrone sulphate was 204 pmol/ml in the umbilical artery and 145 pmol/ml in the umbilical vein. At Caesarean section, on the other hand, the maternal peripheral serum level of oestrone sulphate averaged 362 pmol/ml. The mean serum levels of oestrone sulphate wre 90.7 pmol/ml and 171 pmol/ml in the umbilical artery and umbilical vein, respectively. These results suggest a maternal origin of oestrone sulphate in pregnancy, with fluctuations in the levels being of interest in relation to labour pain. (author)

Agrin and Perlecan Mediate Tumorigenic Processes in Oral Squamous Cell Carcinoma

Science.gov (United States)

Kawahara, Rebeca; Granato, Daniela C.; Carnielli, Carolina M.; Cervigne, Nilva K.; Oliveria, Carine E.; Martinez, César A. R.; Yokoo, Sami; Fonseca, Felipe P.; Lopes, Marcio; Santos-Silva, Alan R.; Graner, Edgard; Coletta, Ricardo D.; Leme, Adriana Franco Paes

2014-01-01

Oral squamous cell carcinoma is the most common type of cancer in the oral cavity, representing more than 90% of all oral cancers. The characterization of altered molecules in oral cancer is essential to understand molecular mechanisms underlying tumor progression as well as to contribute to cancer biomarker and therapeutic target discovery. Proteoglycans are key molecular effectors of cell surface and pericellular microenvironments, performing multiple functions in cancer. Two of the major basement membrane proteoglycans, agrin and perlecan, were investigated in this study regarding their role in oral cancer. Using real time quantitative PCR (qRT-PCR), we showed that agrin and perlecan are highly expressed in oral squamous cell carcinoma. Interestingly, cell lines originated from distinct sites showed different expression of agrin and perlecan. Enzymatically targeting chondroitin sulfate modification by chondroitinase, oral squamous carcinoma cell line had a reduced ability to adhere to extracellular matrix proteins and increased sensibility to cisplatin. Additionally, knockdown of agrin and perlecan promoted a decrease on cell migration and adhesion, and on resistance of cells to cisplatin. Our study showed, for the first time, a negative regulation on oral cancer-associated events by either targeting chondroitin sulfate content or agrin and perlecan levels. PMID:25506919

The hypobranchial mucin of the whelk Buccinum undatum L. The polysaccharide sulphate component.

Science.gov (United States)

Hunt, S; Jevons, F R

1966-02-01

1. A polysaccharide sulphate has been isolated from the hypobranchial mucin of the whelk Buccinum undatum. 2. The molecular weight of this polysaccharide, which is a glucan carrying one ester sulphate group per monosaccharide residue, is 1.7x10(5). 3. Some investigations bearing on the location of the ester sulphate groups are reported. 4. The viscosity of the whole mucin has been shown to depend mainly on the glucan sulphate.

Microwave assisted synthesis of nano sized sulphate doped hydroxyapatite

Energy Technology Data Exchange (ETDEWEB)

Alshemary, Ammar Z.; Goh, Yi-Fan; Akram, Muhammad; Razali, Ili Rabihah [Department of Chemistry, Faculty of Science, Universiti Teknologi Malaysia, 81310 UTM Skudai, Johor Darul Ta’zim (Malaysia); Abdul Kadir, Mohammed Rafiq [Medical Implant Technology Group, Faculty of Biomedical Engineering and Health Science, Universiti Teknologi Malaysia,81310 UTM Skudai, Johor Darul Ta’zim (Malaysia); Hussain, Rafaqat, E-mail: rafaqat@kimia.fs.utm.my [Ibnu Sina Institute for Fundamental Science Studies, Universiti Teknologi Malaysia, 81310 UTM Skudai, Johor DarulTa’zim (Malaysia)

2013-06-01

Highlights: ► Phase pure nano-sized sulphur doped hydroxyapatite has been synthesized. ► TEM analysis confirmed formation of needle shaped structure. ► Lattice parameters and cell volume increased with increase in sulphate doping. ► Crystallite size decreased as sulphate content inside the structure increased. ► Degree of crystallinity decreased with increase in sulphate substitution. - Abstract: Inorganic sulphate is required by all mammalian cells to function properly, it is the fourth most abundant anion in the human plasma. Sulphate ions are the major source of sulphur which is considered an important element for sustenance of life as it is present in the essential amino and is required by cells to function properly. In this study we have successfully substituted sulphate ions (SO{sub 4}{sup 2−}) into hydroxyapatite (Ca{sub 10}(PO{sub 4}){sub 6−x}(SO{sub 4}){sub x}(OH){sub 2−x}) lattice via ion exchange process with phosphate group. Concentration of SO{sub 4}{sup 2−} ions was varied between X = 0.05–0.5, using (Ca (NO{sub 3}){sub 2}·4H{sub 2}O), ((NH{sub 4}){sub 2}HPO{sub 4}) and (Na{sub 2}SO{sub 4}) as starting materials. X-ray diffraction (XRD), Fourier transform IR spectroscopy (FTIR), showed that the substitution of SO{sub 4}{sup 2−} ions into the lattice resulted in peak broadening and reduced peak height due to the amorphous nature and reduced crystallinity of the resulting HA powder. Transmission electron microscopy (TEM) and field emission electron microscopy (FESEM) analysis confirmed the formation of needle shaped particles of 41 nm size with homogenous and uniform distribution of element within the HA structure.

Characteristics and antioxidant of Ulva intestinalis sulphated polysaccharides extracted with different solvents.

Science.gov (United States)

Peasura, Napassorn; Laohakunjit, Natta; Kerdchoechuen, Orapin; Wanlapa, Sorada

2015-11-01

Ulva intestinalis, a tubular green seaweed, is a rich source of nutrient, especially sulphated polysaccharides. Sulphated polysaccharides from U. intestinalis were extracted with distilled water, 0.1N HCl, and 0.1N NaOH at 80°C for 1, 3, 6, 12, and 24h to study the effect of the extraction solvent and time on their chemical composition and antioxidant activity. Different types of solvents and extraction time had a significant influence on the chemical characteristics and antioxidant activity (pMonosaccharide composition and FT-IR spectra analyses revealed that sulphated polysaccharides from all solvent extractions have a typical sugar backbone (glucose, rhamnose, and sulphate attached at C-2 or C-3 of rhamnose). Sulphated polysaccharides extracted with acid exhibited greater antioxidant activity than did those extracted with distilled water and alkali. The results indicated that solvent extraction could be an efficacious method for enhancing antioxidant activity by distinct molecular weight and chemical characteristic of sulphated polysaccharides. Copyright © 2015 Elsevier B.V. All rights reserved.

Quantitation and identification of methanogens and sulphate reducers in Olkiluoto groundwater

International Nuclear Information System (INIS)

Bomberg, M.; Nyyssoenen, M.; Itaevaara, M.

2010-08-01

The GEOFUNC Project focuses on the microbiology connected to safety and risk assessment of the final disposal of high radioactive nuclear waste. Methanogenic archaea and sulphate reducing bacteria are significant groups of microorganisms in anaerobic environments, and are of crucial concern for the safe long term storage of nuclear waste in deep bedrock. The sulphate reducing bacteria are able to produce sulphide which may cause corrosion of the copper in the radioactive waste storage capsules. Methanogens, on the other hand, may produce quantities of methane from various organic carbon compounds, CO 2 and H 2 . Methane may both serve as carbon source for methanotrophic microbial groups, and may also cause mobilization of radionuclides, as a result of gas discharge through fractures in the bedrock. The transition zones between the sulphate rich and methane rich waters are locations for microbial processes where the methane may serve as carbon source for sulphate reducing bacteria, which in turn would produce corrosive sulphides. It has been estimated that only 1-10 % of all the microorganisms present in the environment can be isolated and cultivated. Uncultured microorganisms can be identified and their numbers in the environment quantified by identification of specific marker genes that are essential for their functions by use of molecular methods. Methanogens, for example, can be identified by their genes for methyl coenzyme M reductase (mcrA), which is an essential enzyme involved in the production of methane. The mcrA is specifically present only in methanogenic archaea. Sulphate reducers are identified by their dissimilatory sulphite reductase genes (dsrB), which are present in and essential for all microorganisms performing dissimilatory sulphate reducing. In the GEOFUNC project, a quantitative PCR method (qPCR) was developed for the detection of methanogens and sulphate reducers. This method is based on specific quantitative detection of marker genes

Inter vs. intraglycosidic acetal linkages control sulfation pattern in semi-synthetic chondroitin sulfate.

Science.gov (United States)

Laezza, Antonio; De Castro, Cristina; Parrilli, Michelangelo; Bedini, Emiliano

2014-11-04

Microbial-sourced unsulfated chondroitin could be converted into chondroitin sulfate (CS) polysaccharide by a multi-step strategy relying upon benzylidenation and acetylation reactions as key-steps for its regioselective protection. By conducting the two reactions one- or two-pots, CSs with different sulfation patterns could be obtained at the end of the semi-synthesis. In particular, a CS polysaccharide possessing sulfate groups randomly distributed between positions 4 and 6 of N-acetyl-galactosamine (GalNAc) units could be obtained through the two-pots route, whereas the one-pot pathway allowed an additional sulfation at position 3 of some glucuronic acid (GlcA) units. This difference was ascribed to the stabilization of a labile interglycosidic benzylidene acetal involving positions O-3 and O-6 of some GlcA and GalNAc, respectively, when the benzylidene-acetylation reactions were conducted in a one-pot fashion. Isolation and characterization of a polysaccharide intermediate showing interglycosidic acetal moieties was accomplished. Copyright © 2014 Elsevier Ltd. All rights reserved.

Control of extracellular matrix assembly by syndecan-2 proteoglycan

DEFF Research Database (Denmark)

Klass, C M; Couchman, J R; Woods, A

2000-01-01

Extracellular matrix (ECM) deposition and organization is maintained by transmembrane signaling and integrins play major roles. We now show that a second transmembrane component, syndecan-2 heparan sulfate proteoglycan, is pivotal in matrix assembly. Chinese Hamster Ovary (CHO) cells were stably...... to rearrange laminin or fibronectin substrates into fibrils and to bind exogenous fibronectin. Transfection of activated alphaIIbalphaLdeltabeta3 integrin into alpha(5)-deficient CHO B2 cells resulted in reestablishment of the previously lost fibronectin matrix. However, cotransfection of this cell line with S...

Dose-response testing with nickel sulphate using the TRUE test in nickel-sensitive individuals. Multiple nickel sulphate patch-test reactions do not cause an 'angry back'

DEFF Research Database (Denmark)

Andersen, Klaus Ejner; Lidén, C; Hansen, J

1993-01-01

The aim of this study was to employ the TRUE test assay to confirm the presence or absence of the 'angry back' phenomenon, i.e. that a strong positive patch-test reaction heightens adjacent patch-test response. In addition, we wished to establish the dose-response relationship for nickel sulphate...... back' phenomenon was not apparent in this study, as the spill-over effect was not statistically significant. Strong reactions to high concentrations of nickel sulphate did not enhance the response to adjacent lower concentrations of nickel sulphate....

Corrosion Performance of Inconel 625 in High Sulphate Content

Science.gov (United States)

Ismail, Azzura

2016-05-01

Inconel 625 (UNS N06625) is a type of nickel-chromium-molybdenum alloy with excellent corrosion resistance in a wide range of corrosive media, being especially resistant to pitting and crevice corrosion. However, in aggressive environment, Inconel 625 will suffer corrosion attack like other metals. This research compared the corrosion performance of Inconel 625 when exposed to higher sulphate content compared to real seawater. The results reveal that Inconel 625 is excellent in resist the corrosion attack in seawater. However, at increasing temperature, the corrosion resistance of this metal decrease. The performance is same in seawater with high sulphate content at increasing temperature. It can be concluded that sulphate promote perforation on Inconel 625 and become aggressive agents that accelerate the corrosion attack.

Agrin is a major heparan sulfate proteoglycan in the human glomerular basement membrane

NARCIS (Netherlands)

Groffen, Alexander J.; Ruegg, Markus A.; Dijkman, Henri; Van De Velden, Thea J.; Buskens, Carin A.; Van Den Born, Jacob; Assmann, Karel J.; Monnens, Leo A.; Veerkamp, Jacques H.; Van Den Heuvel, Lambert P.

Agrin is a heparan sulfate proteoglycan (HSPG) that is highly concentrated in the synaptic basal lamina at the neuromuscular junction (NMJ). Agrin-like immunoreactivity is also detected outside the NMJ. Here we show that agrin is a major HSPG component of the human glomerular basement membrane

Comparison of Glucosamine-Chondroitin Sulfate with and without Methylsulfonylmethane in Grade I-II Knee Osteoarthritis: A Double Blind Randomized Controlled Trial.

Science.gov (United States)

Lubis, Andri M T; Siagian, Carles; Wonggokusuma, Erick; Marsetyo, Aldo F; Setyohadi, Bambang

2017-04-01

Glucosamine, chondroitinsulfate are frequently used to prevent further joint degeneration in osteoarthritis (OA). Methylsulfonylmethane (MSM) is a supplement containing organic sulphur and also reported to slow anatomical joint progressivity in the knee OA. The MSM is often combined with glucosamine and chondroitin sulfate. However, there are controversies whether glucosamine-chondroitin sulfate or their combination with methylsulfonylmethane could effectively reduce pain in OA. This study is aimed to compare clinical outcome of glucosamine-chondroitin sulfate (GC), glucosamine-chondroitin sulfate-methylsulfonylmethane (GCM), and placeboin patients with knee osteoarthritis (OA) Kellgren-Lawrence grade I-II. a double blind, randomized controlled clinical trial was conducted on 147 patients with knee OA Kellgren-Lawrence grade I-II. Patients were allocated by permuted block randomization into three groups: GC (n=49), GCM (n=50), or placebo (n=48) groups. GC group received 1500 mg of glucosamine + 1200 mg of chondroitin sulfate + 500 mg of saccharumlactis; GCM group received 1500 mg of glucosamine + 1200 mg of chondroitin sulfate + 500 mg of MSM; while placebo group received three matching capsules of saccharumlactis. The drugs were administered once daily for 3 consecutive months VAS and WOMAC scores were measured before treatment, then at 4th, 8th and 12th week after treatment. on statistical analysis it was found that at the 12th week, there are significant difference between three treatment groups on the WOMAC score (p=0.03) and on the VAS score (p=0.004). When analyzed between weeks, GCM treatment group was found statistically significant on WOMAC score (p=0.01) and VAS score (p<0.001). Comparing the score difference between weeks, WOMAC score analysis showed significant difference between GC, GCM, and placebo in week 4 (p=0.049) and week 12 (p=0.01). In addition, VAS score also showed significant difference between groups in week 8 (p=0.006) and week 12 (p<0

Glycosaminoglycan composition of PC12 pheochromocytoma cells: a comparison with PC12D cells, a new subline of PC12 cells

Energy Technology Data Exchange (ETDEWEB)

Katoh-Semba, R.; Oohira, A.; Sano, M.; Watanabe, K.; Kitajima, S.; Kashiwamata, S.

1989-03-01

PC12D cells, a new subline of conventional PC12 cells, respond not only to nerve growth factor but also to cyclic AMP by extending their neurites. These cells are flat in shape and are similar in appearance to PC12 cells that have been treated with nerve growth factor for a few days. In both cell lines, we have characterized the glycosaminoglycans, the polysaccharide moieties of proteoglycans, which are believed to play an important role in cell adhesion and in cell morphology. Under the present culture conditions, only chondroitin sulfate was detected in the media from PC12 and PC12D cells, whereas both chondroitin sulfate and heparan sulfate were found in the cell layers. The levels of cell-associated heparan sulfate and chondroitin sulfate were about twofold and fourfold higher in PC12D cells than in PC12 cells, respectively. Compared to PC12 cells, the amounts of (/sup 35/S)sulfate incorporated for 48 h into chondroitin sulfate were twofold lower but those into heparan sulfate were 35% higher in PC12D cells. The amount of chondroitin sulfate released by PC12D cells into the medium was about a half of that released by PC12 cells. The ratio of (/sup 35/S)sulfate-labeled heparan sulfate to chondroitin sulfate was 6.2 in PC12D cells and 2.2 in PC12 cells. These results suggest that there may be some correlation between the increase in content of glycosaminoglycans and the change in cell morphology, which is followed by neurite outgrowth.

Carbohydrate as covalent crosslink in human inter-alpha-trypsin inhibitor

DEFF Research Database (Denmark)

Jessen, T E; Faarvang, K L; Ploug, M

1988-01-01

The primary structure of inter-alpha-trypsin inhibitor is partially elucidated, but controversy about the construction of the polypeptide backbone still exists. We present evidence suggesting that inter-alpha-trypsin inhibitor represents a novel plasma protein structure with two separate polypept...... polypeptide chains covalently crosslinked only by carbohydrate (chondroitin sulphate)....

Ablation of NG2 proteoglycan leads to deficits in brown fat function and to adult onset obesity.

Directory of Open Access Journals (Sweden)

Yunchao Chang

Full Text Available Obesity is a major health problem worldwide. We are studying the causes and effects of obesity in C57Bl/6 mice following genetic ablation of NG2, a chondroitin sulfate proteoglycan widely expressed in progenitor cells and also in adipocytes. Although global NG2 ablation delays early postnatal adipogenesis in mouse skin, adult NG2 null mice are paradoxically heavier than wild-type mice, exhibiting larger white fat deposits. This adult onset obesity is not due to NG2-dependent effects on CNS function, since specific ablation of NG2 in oligodendrocyte progenitors yields the opposite phenotype; i.e. abnormally lean mice. Metabolic analysis reveals that, while activity and food intake are unchanged in global NG2 null mice, O(2 consumption and CO(2 production are decreased, suggesting a decrease in energy expenditure. Since brown fat plays important roles in regulating energy expenditure, we have investigated brown fat function via cold challenge and high fat diet feeding, both of which induce the adaptive thermogenesis that normally occurs in brown fat. In both tests, body temperatures in NG2 null mice are reduced compared to wild-type mice, indicating a deficit in brown fat function in the absence of NG2. In addition, adipogenesis in NG2 null brown pre-adipocytes is dramatically impaired compared to wild-type counterparts. Moreover, mRNA levels for PR domain containing 16 (PRDM16 and peroxisome proliferator-activated receptor γ coactivator (PGC1-α, proteins important for brown adipocyte differentiation, are decreased in NG2 null brown fat deposits in vivo and NG2 null brown pre-adipocytes in vitro. Altogether, these results indicate that brown fat dysfunction in NG2 null mice results from deficits in the recruitment and/or development of brown pre-adipocytes. As a consequence, obesity in NG2 null mice may occur due to disruptions in brown fat-dependent energy homeostasis, with resulting effects on lipid storage in white adipocytes.

Study of the sulphate expansion phenomenon in concrete: behaviour of the cemented radioactive wastes containing sulphate

International Nuclear Information System (INIS)

Li, Guanshu

1994-01-01

Sulphate attack is one of the major degradation processes of concrete. It is especially important in storing cemented radioactive wastes containing sulphate. In this thesis, we have thoroughly investigated the degradation mechanisms of cemented radioactive wastes by sulphate. The CaO-Al 2 O 3 -SO 3 -H 2 O systems with and without alkalis are studied. For the system without alkalis, experimental results show that it is the formation of a secondary ettringite under external water supply by steric effect that causes the expansion. For the system with alkalis, the ettringite does not appear while a new mineral called 'U', a sodium-substituted AFm phase is detected. This phase is shown to be responsible for the expansion and destruction of the specimens. The conditions for the formation, the product of solubility and many means of its synthesis are discussed, and a complete list of the inter-reticular distances file is given. The behaviour of the different types of cemented wastes containing sulphate are then studied with a special focus on the U phase on entity which was heretofore very little understood. The following three hypothetical mechanisms of sulphate expansion are proposed: the formation of the secondary U phase, the transformation of the U phase to the ettringite and the topochemical hydration of thenardite into mirabilite. Experiments on a simplified system have demonstrated clearly that the formation of the secondary U phase can induce enormous expansion by steric effect, this justifying the first assumption. Simulation by the mass and volume balances is carried out thereafter and enables us to estimate the expansion induced by the formation of the secondary U phase in the cemented wastes. The second assumption is also well verified by a series of leaching tests in different solutions on mixtures containing the U phase. On the basis of the analysis of the specimens under leaching, it has been assumed that the expansion is associated with the

Chinese hamster ovary cell mutants defective in heparan sulfate biosynthesis

International Nuclear Information System (INIS)

Bame, K.J.; Kiser, C.S.; Esko, J.D.

1987-01-01

The authors have isolated Chinese hamster ovary cell mutants defective in proteoglycan synthesis by radiographic screening for cells unable to incorporate 35 SO 4 into acid-precipitable material. Some mutants did not incorporate 35 SO 4 into acid-precipitable material, whereas others incorporated about 3-fold less radioactivity. HPLC anion exchange chromatographic analysis of radiolabelled glycosaminoglycans isolated from these mutants revealed many are defective in heparan sulfate biosynthesis. Mutants 803 and 677 do not synthesize heparan sulfate, although they produce chondroitin sulfate: strain 803 makes chondroitin sulfate normally, whereas 677 overaccumulates chondroitin sulfate by a factor of three. These mutants fall into the same complementation group, suggesting that the mutations are allelic. A second group of heparan sulfate biosynthetic mutants, consisting of cell lines 625, 668 and 679, produce undersulfated heparan sulfate and normal chondroitin sulfate. Treatment of the chains with nitrous acid should determine the position of the sulfate groups along the chain. These mutants may define a complementation group that is defective in the enzymes which modify the heparan sulfate chain. To increase the authors repertoire of heparan sulfate mutants, they are presently developing an in situ enzyme assay to screen colonies replica plated on filter discs for sulfotransferase defects

The effect of water on the sulphation of limestone

Energy Technology Data Exchange (ETDEWEB)

Chunbo Wang; Lufei Jia; Yewen Tan; E.J. Anthony [North China Electric Power University, Baoding City (China). School of Energy and Power Engineering

2010-09-15

A series of tests was conducted in a thermogravimetric analyzer (TGA) to study the sulphation behaviour of limestone in the presence of water over the temperature range of 800-850{sup o}C. Four different Canadian limestones, all with a particle size range of 75-425 {mu}m, were sulphated using a synthetic flue gas with a composition of 15% CO{sub 2}, 3% O{sub 2}, 0% or 10% H{sub 2}O, 1750 ppm SO{sub 2} and the balance N{sub 2}. Water was shown to have a significant promotional effect on sulphation, especially in the diffusion-controlled stage. However, the effect of water during the kinetic-controlled stage appeared to be much less pronounced. Based on these results, it is proposed that the presence of water leads to the transient formation of Ca(OH){sub 2} as an intermediate, which in turn reacts with SO{sub 2} at a faster rate than CaO does. Alternatively stated, it appears that H{sub 2}O acts as catalyst for the sulphation reaction of CaO. 30 refs., 8 figs., 3 tabs.

The action of diazoxide and minoxidil sulphate on rat blood vessels: a comparison with cromakalim.

Science.gov (United States)

Newgreen, D. T.; Bray, K. M.; McHarg, A. D.; Weston, A. H.; Duty, S.; Brown, B. S.; Kay, P. B.; Edwards, G.; Longmore, J.; Southerton, J. S.

1990-01-01

1. The actions of diazoxide and minoxidil sulphate have been compared with those of cromakalim in rat aorta and portal vein. 2. Diazoxide and minoxidil sulphate hyperpolarized the rat portal vein in a similar manner to cromakalim. 3. Cromakalim, diazoxide and minoxidil sulphate increased 42K and 86Rb efflux from rat portal vein, although minoxidil sulphate had only a small effect on 86Rb efflux. 4. Cromakalim, diazoxide and minoxidil sulphate increased 42K efflux from rat aorta but only cromakalim and diazoxide increased 86Rb efflux from this tissue. 5. Glibenclamide inhibited the relaxant actions of cromakalim, diazoxide and minoxidil sulphate on rat aorta and the increase in 42K efflux produced by these agents in this tissue. 6. Diazoxide relaxed an 80 mM KCl-induced contraction of rat aorta, whilst cromakalim and minoxidil sulphate were without effect. 7. Cromakalim, diazoxide and minoxidil sulphate had no effect on cyclic AMP or cyclic GMP concentrations in rat aorta. 8. It is concluded that diazoxide and minoxidil sulphate like cromakalim exhibit K+ channel opening properties in vascular smooth muscle. Diazoxide exerts an additional inhibitory action not related to the production of cyclic AMP or cyclic GMP. The action of minoxidil sulphate may be primarily located at a K+ channel which is relatively impermeable to 86Rb. PMID:2167738

Proteoglycans as potential biomarkers in odontogenic tumors

Science.gov (United States)

Gómez-Herrera, Zaira; Molina-Frechero, Nelly; Damián-Matsumura, Pablo; Bologna-Molina, Ronell

2018-01-01

Proteoglycans (PGs) are essential for normal cellular development; however, alterations of their concentrations can promote tumor growth. To date, a limited number of studies report the presence of PGs in odontogenic tumors (OTs); therefore, the main purpose of this work is to gather the information published on the study of PGs. The search reported 26 articles referring to the presence of different PGs in distinct OTs from 1999 to May 2017. PGs seem to play an important role during OTs’ development as they are involved in several tumor processes; however, the number of reports on the study of these molecules is low. Thus, more studies are necessary in order to gain a better understanding of the underlying pathophysiology of OTs. PMID:29731564

Chondroitin sulfate A-adhering Plasmodium falciparum-infected erythrocytes express functionally important antibody epitopes shared by multiple variants

DEFF Research Database (Denmark)

Barfod, Lea; Dobrilovic, Tina; Magistrado, Pamela

2010-01-01

to chondroitin sulfate A in the intervillous space. Although interclonal variation of the var2csa gene is lower than that among var genes in general, VAR2CSA-specific Abs appear to target mainly polymorphic epitopes. This has raised doubts about the feasibility of VAR2CSA-based vaccines. We used eight human......) and recombinant VAR2CSA and interfered with IE and/or VAR2CSA binding to chondroitin sulfate A. Pair-wise mAb combinations were more inhibitory than single mAbs, and all of the mAbs together was the most efficient combination. Each mAb could opsonize IEs for phagocytosis, and a combination of the eight m...

Human mast cell neutral proteases generate modified LDL particles with increased proteoglycan binding.

Science.gov (United States)

Maaninka, Katariina; Nguyen, Su Duy; Mäyränpää, Mikko I; Plihtari, Riia; Rajamäki, Kristiina; Lindsberg, Perttu J; Kovanen, Petri T; Öörni, Katariina

2018-04-13

Subendothelial interaction of LDL with extracellular matrix drives atherogenesis. This interaction can be strengthened by proteolytic modification of LDL. Mast cells (MCs) are present in atherosclerotic lesions, and upon activation, they degranulate and release a variety of neutral proteases. Here we studied the ability of MC proteases to cleave apoB-100 of LDL and affect the binding of LDL to proteoglycans. Mature human MCs were differentiated from human peripheral blood-derived CD34 + progenitors in vitro and activated with calcium ionophore to generate MC-conditioned medium. LDL was incubated in the MC-conditioned medium or with individual MC proteases, and the binding of native and modified LDL to isolated human aortic proteoglycans or to human atherosclerotic plaques ex vivo was determined. MC proteases in atherosclerotic human coronary artery lesions were detected by immunofluorescence and qPCR. Activated human MCs released the neutral proteases tryptase, chymase, carboxypeptidase A3, cathepsin G, and granzyme B. Of these, cathepsin G degraded most efficiently apoB-100, induced LDL fusion, and enhanced binding of LDL to isolated human aortic proteoglycans and human atherosclerotic lesions ex vivo. Double immunofluoresence staining of human atherosclerotic coronary arteries for tryptase and cathepsin G indicated that lesional MCs contain cathepsin G. In the lesions, expression of cathepsin G correlated with the expression of tryptase and chymase, but not with that of neutrophil proteinase 3. The present study suggests that cathepsin G in human atherosclerotic lesions is largely derived from MCs and that activated MCs may contribute to atherogenesis by enhancing LDL retention. Copyright © 2018 Elsevier B.V. All rights reserved.

Magnesium sulphate and other anticonvulsants for women with pre-eclampsia.

Science.gov (United States)

Duley, Lelia; Gülmezoglu, A Metin; Henderson-Smart, David J; Chou, Doris

2010-11-10

Eclampsia, the occurrence of a seizure (fit) in association with pre-eclampsia, is rare but potentially life-threatening. Magnesium sulphate is the drug of choice for treating eclampsia. This review assesses its use for preventing eclampsia. To assess the effects of magnesium sulphate, and other anticonvulsants, for prevention of eclampsia. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (4 June 2010), and the Cochrane Central Register of Controlled Trials Register (The Cochrane Library 2010, Issue 3). Randomised trials comparing anticonvulsants with placebo or no anticonvulsant, or comparisons of different drugs, for pre-eclampsia. Two authors assessed trial quality and extracted data independently. We included 15 trials. Six (11,444 women) compared magnesium sulphate with placebo or no anticonvulsant: magnesium sulphate more than a halved the risk of eclampsia (risk ratio (RR) 0.41, 95% confidence interval (CI) 0.29 to 0.58; number needed to treat for an additional beneficial outcome (NNTB) 100, 95% CI 50 to 100), with a non-significant reduction in maternal death (RR 0.54, 95% CI 0.26 to 1.10) but no clear difference in serious maternal morbidity (RR 1.08, 95% CI 0.89 to 1.32). It reduced the risk of placental abruption (RR 0.64, 95% CI 0.50 to 0.83; NNTB 100, 95% CI 50 to 1000), and increased caesarean section (RR 1.05, 95% CI 1.01 to 1.10). There was no clear difference in stillbirth or neonatal death (RR 1.04, 95% CI 0.93 to 1.15). Side effects, primarily flushing, were more common with magnesium sulphate (24% versus 5%; RR 5.26, 95% CI 4.59 to 6.03; number need to treat for an additional harmful outcome (NNTH) 6, 95% CI 5 to 6).Follow-up was reported by one trial comparing magnesium sulphate with placebo: for 3375 women there was no clear difference in death (RR 1.79, 95% CI 0.71 to 4.53) or morbidity potentially related to pre-eclampsia (RR 0.84, 95% CI 0.55 to 1.26) (median follow-up 26 months); for 3283 children exposed in utero

Sequencing of chondroitin sulfate oligosaccharides using a novel exolyase from a marine bacterium that degrades hyaluronan and chondroitin sulfate/dermatan sulfate.

Science.gov (United States)

Wang, Wenshuang; Cai, Xiaojuan; Han, Naihan; Han, Wenjun; Sugahara, Kazuyuki; Li, Fuchuan

2017-11-09

Glycosaminoglycans (GAGs) are a family of chemically heterogeneous polysaccharides that play important roles in physiological and pathological processes. Owing to the structural complexity of GAGs, their sophisticated chemical structures and biological functions have not been extensively studied. Lyases that cleave GAGs are important tools for structural analysis. Although various GAG lyases have been identified, exolytic lyases with unique enzymatic property are urgently needed for GAG sequencing. In the present study, a putative exolytic GAG lyase from a marine bacterium was recombinantly expressed and characterized in detail. Since it showed exolytic lyase activity toward hyaluronan (HA), chondroitin sulfate (CS), and dermatan sulfate (DS), it was designated as HCDLase. This novel exolyase exhibited the highest activity in Tris-HCl buffer (pH 7.0) at 30°C. Especially, it showed a specific activity that released 2-aminobenzamide (2-AB)-labeled disaccharides from the reducing end of 2-AB-labeled CS oligosaccharides, which suggest that HCDLase is not only a novel exolytic lyase that can split disaccharide residues from the reducing termini of sugar chains but also a useful tool for the sequencing of CS chains. Notably, HCDLase could not digest 2-AB-labeled oligosaccharides from HA, DS, or unsulfated chondroitin, which indicated that sulfates and bond types affect the catalytic activity of HCDLase. Finally, this enzyme combined with CSase ABC was successfully applied for the sequencing of several CS hexa- and octasaccharides with complex structures. The identification of HCDLase provides a useful tool for CS-related research and applications. © 2017 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Glucosamine:chondroitin or ginger root extract have little effect on articular cartilage in swine

Science.gov (United States)

Sows are culled at a high rate from breeding herds due to musclo-skeletal problems and lameness. Research in our laboratory has shown that even first-parity sows have significant amounts of osteochondritic lesions of their articular cartilage. Glusoamine chondroitin and ginger root extract have both...

Performance of cement solidification with barium for high activity liquid waste including sulphate

International Nuclear Information System (INIS)

Waki, Toshikazu; Yamada, Motoyuki; Horikawa, Yoshihiko; Kaneko, Masaaki; Saso, Michitaka; Haruguchi, Yoshiko; Yamashita, Yu; Sakai, Hitoshi

2009-01-01

The target liquid waste to be solidified is generated from PWR primary loop spent resin treatment with sulphate acid, so, its main constituent is sodium sulphate and the activity of this liquid is relatively high. Waste form of this liquid waste is considered to be a candidate for the subsurface disposal. The disposed waste including sulphate is anticipated to rise a concentration of sulphate ion in the ground water around the disposal facility and it may cause degradation of materials such as cement and bentonite layer and comprise the disposal facility. There could be two approaches to avoid this problem, the strong design of the disposal facility and the minimization of sulphaste ion migration from the solidified waste. In this study, the latter approach was examined. In order to keep the low concentration of sulphate ion in the ground water, it is effective to make barium sulphate by adding barium compound into the liquid waste in solidification. However, adding equivalent amount of barium compound with sulphate ion causes difficulty of mixing, because production of barium sulphate causes high viscosity. In this study, mixing condition after and before adding cement into the liquid waste was estimated. The mixing condition was set with consideration to keep anion concentration low in the ground water and of mixing easily enough in practical operation. Long term leaching behavior of the simulated solidified waste was also analyzed by PHREEQC. And the concentration of the constitution affected to the disposal facility was estimated be low enough in the ground water. (author)

Preparation and characterization of hydroxyapatite/chondroitin sulfate composites by biomimetic synthesis

International Nuclear Information System (INIS)

Xiao Xiufeng; He Dan; Liu Fang; Liu Rongfang

2008-01-01

Based on the principles of biomineralization, flakelike hydroxyapatite/chondroitin sulfate composites were synthesized through biomimetic method using Ca(NO 3 ) 2 .4H 2 O and (NH 4 ) 3 PO 4 .3H 2 O as reagents and chondroitin sulfate as template. The crystalline phase, microstructure, chemical composition, morphology and thermal behavior of the composites obtained in the experiment were characterized by means of X-ray diffraction (XRD), Fourier transform infrared spectroscope (FTIR), transmission electron microscope (TEM), Thermogravimetry-Differential thermal analyzer (TG-DTA) and Elemental analyzer, respectively. The interaction between the functional groups of ChS and HA was investigated by electrical conductivity and UV-vis spectrum. The results demonstrate that the as-prepared powders with small amount of carbonate have the component similar to human bone. It can be concluded that the nucleation and growth of HA crystals occurred through the chemical interactions between the HA crystals and preorganized functional groups of the ChS template. Furthermore, the concentration of ChS significantly affects the morphology of the composites. Short fiberlike crystals could be obtained at a low concentration of ChS, but flakelike crystals could be synthesized using a high concentration (≥0.5 wt%) of ChS as template

Preparation and characterization of hydroxyapatite/chondroitin sulfate composites by biomimetic synthesis

Energy Technology Data Exchange (ETDEWEB)

Xiao Xiufeng; He Dan; Liu Fang [College of Chemistry and Materials Science, Fujian Normal University, Fujian, Fuzhou 350007 (China); Liu Rongfang [College of Chemistry and Materials Science, Fujian Normal University, Fujian, Fuzhou 350007 (China)], E-mail: rfliu@vip.sina.com

2008-12-20

Based on the principles of biomineralization, flakelike hydroxyapatite/chondroitin sulfate composites were synthesized through biomimetic method using Ca(NO{sub 3}){sub 2}.4H{sub 2}O and (NH{sub 4}){sub 3}PO{sub 4}.3H{sub 2}O as reagents and chondroitin sulfate as template. The crystalline phase, microstructure, chemical composition, morphology and thermal behavior of the composites obtained in the experiment were characterized by means of X-ray diffraction (XRD), Fourier transform infrared spectroscope (FTIR), transmission electron microscope (TEM), Thermogravimetry-Differential thermal analyzer (TG-DTA) and Elemental analyzer, respectively. The interaction between the functional groups of ChS and HA was investigated by electrical conductivity and UV-vis spectrum. The results demonstrate that the as-prepared powders with small amount of carbonate have the component similar to human bone. It can be concluded that the nucleation and growth of HA crystals occurred through the chemical interactions between the HA crystals and preorganized functional groups of the ChS template. Furthermore, the concentration of ChS significantly affects the morphology of the composites. Short fiberlike crystals could be obtained at a low concentration of ChS, but flakelike crystals could be synthesized using a high concentration ({>=}0.5 wt%) of ChS as template.

Purification, structural characterization and anticoagulant properties of fucosylated chondroitin sulfate isolated from Holothuria mexicana.

Science.gov (United States)

Mou, Jiaojiao; Wang, Cong; Li, Wenjing; Yang, Jie

2017-05-01

A novel fucosylated chondroitin sulfate (HmG) was isolated from sea cucumber Holothuria mexicana, the structure of which was characterized by monosaccharide composition, disaccharide composition, IR, 1 H and 13 C NMR spectrum, additionally with two dimensional NMR spectrum of degraded HmG (DHmG). The backbone of HmG was identified as chondroitin 6-O sulfate, while the major O-4 sulfated fucose branches linked to O-3 position of glucuronic acid in almost every disaccharide unit. The anticoagulant activities of HmG and DHmG were assessed and compared with heparin and low molecular weight heparin. The results indicated that HmG and DHmG both could significantly prolong the activated partial thrombo-plastin time, and the properties were well related to its molecular weight. DHmG showed similar anticoagulant properties to low molecular weight heparin with less bleeding risks, making it a safer anticoagulant drug. Copyright © 2017 Elsevier B.V. All rights reserved.

Comparison of the effects of sodium hyaluronate-chondroitin sulphate and corticosteroid in the treatment of lateral epicondylitis: a prospective randomized trial.

Science.gov (United States)

Tosun, Haci Bayram; Gumustas, Seyitali; Agir, Ismail; Uludag, Abuzer; Serbest, Sancar; Pepele, Demet; Ertem, Kadir

2015-09-01

Hyaluronic acid and glycosaminoglycans have shown positive effects in improving lateral epicondylitis and other tendinosis conditions. Therefore, we designed a prospective, randomized study to compare the effects of a combined sodium hyaluronate and chondroitin sulfate (HA + CS) injection versus a triamcinolone injection in the treatment of lateral epicondylitis. In total, 57 consecutive patients with clinically diagnosed lateral epicondylitis were divided randomly into two groups. In the HA + CS group, 25 patients received a single injection of a solution containing an HA + CS combination and prilocaine HCl, while the 32 patients in the triamcinolone group received a single injection of a solution of triamcinolone and prilocaine HCl. We evaluated the pain and function outcome measures using the Patient-Rated Tennis Elbow Evaluation (PRTEE) questionnaire at the beginning of the study, and 3 and 6 months after the injection. Additionally, the Minimum Clinically Important Difference values and percentage changes in the PRTEE subscale scores between the assessments were calculated. No serious adverse events were reported throughout the study. The mean pain and function scores for the HA + CS and triamcinolone groups had significantly improved at 3 months, but the mean function scores in the HA + CS group were statistically significantly better when compared to the triamcinolone group. At 6 months, both groups had significantly improved mean pain and function scores, compared to the baseline scores; however, the mean pain and function scores in the 6-month HA + CS treatment group were better than in the 6-month triamcinolone group. The relative change for the mean total score in the HA + CS group was much better when compared with the triamcinolone group, and the HA + CS treatment group showed clinically significant improvement when compared with triamcinolone group at 3 and 6 months. This study supports the idea that for a single injection treatment of patients with

Effect of temperature on sulphate reduction, growth rate and growth yield in five psychrophilic sulphate-reducing bacteria from Arctic sediments

DEFF Research Database (Denmark)

Knoblauch, C.; Jørgensen, BB

1999-01-01

Five psychrophilic sulphate-reducing bacteria (strains ASv26, LSv21, PSv29, LSv54 and LSv514) isolated from Arctic sediments were examined for their adaptation to permanently low temperatures, All strains grew at -1.8 degrees C, the freezing point of sea water, but their optimum temperature...... and T(opt). For strains LSv21 and LSv514, however, growth yields were highest at the lowest temperatures, around 0 degrees C. The results indicate that psychrophilic sulphate-reducing bacteria are specially adapted to permanently low temperatures by high relative growth rates and high growth yields...... at in site conditions....

Complete sulphate removal from neutralised acidic mine drainage with barium carbonate

CSIR Research Space (South Africa)

Swanepoel, H

2012-03-01

Full Text Available -barium-calcium) Desalination process which uses barium salts to further reduce the sulphate concentration to acceptable levels with the added advantage that sulphate removal can be controlled due to the low solubility of BaSO4. This paper reports on the results...

Wnt Signaling Cascades and the Roles of Syndecan Proteoglycans

DEFF Research Database (Denmark)

Pataki, Csilla A; Couchman, John R; Brábek, Jan

2015-01-01

/planar cell polarity and Wnt/calcium signaling. Syndecans are type I transmembrane proteoglycans with a long evolutionary history, being expressed in all Bilateria and in almost all cell types. Both Wnt pathways have been extensively studied over the past 30 years and shown to have roles during development...... and in a multitude of diseases. Although the first evidence for interactions between syndecans and Wnts dates back to 1997, the number of studies connecting these pathways is low, and many open questions remained unanswered. In this review, syndecan's involvement in Wnt signaling pathways as well as some...

Heparan sulfate proteoglycans: structure, protein interactions and cell signaling

Directory of Open Access Journals (Sweden)

Juliana L. Dreyfuss

2009-09-01

Full Text Available Heparan sulfate proteoglycans are ubiquitously found at the cell surface and extracellular matrix in all the animal species. This review will focus on the structural characteristics of the heparan sulfate proteoglycans related to protein interactions leading to cell signaling. The heparan sulfate chains due to their vast structural diversity are able to bind and interact with a wide variety of proteins, such as growth factors, chemokines, morphogens, extracellular matrix components, enzymes, among others. There is a specificity directing the interactions of heparan sulfates and target proteins, regarding both the fine structure of the polysaccharide chain as well precise protein motifs. Heparan sulfates play a role in cellular signaling either as receptor or co-receptor for different ligands, and the activation of downstream pathways is related to phosphorylation of different cytosolic proteins either directly or involving cytoskeleton interactions leading to gene regulation. The role of the heparan sulfate proteoglycans in cellular signaling and endocytic uptake pathways is also discussed.Proteoglicanos de heparam sulfato são encontrados tanto superfície celular quanto na matriz extracelular em todas as espécies animais. Esta revisão tem enfoque nas características estruturais dos proteoglicanos de heparam sulfato e nas interações destes proteoglicanos com proteínas que levam à sinalização celular. As cadeias de heparam sulfato, devido a sua variedade estrutural, são capazes de se ligar e interagir com ampla gama de proteínas, como fatores de crescimento, quimiocinas, morfógenos, componentes da matriz extracelular, enzimas, entreoutros. Existe uma especificidade estrutural que direciona as interações dos heparam sulfatos e proteínas alvo. Esta especificidade está relacionada com a estrutura da cadeia do polissacarídeo e os motivos conservados da cadeia polipeptídica das proteínas envolvidas nesta interação. Os heparam

Burkitt lymphoma expresses oncofetal chondroitin sulfate without being a reservoir for placental malaria sequestration

DEFF Research Database (Denmark)

Agerbaek, Mette Ø; Bento Ayres Pereira, Marina Maria; Clausen, Thomas M

2017-01-01

in other non-malignant tissues and thus VAR2CSA generally facilitates parasite sequestration and accumulation in pregnant women. In this study, we show that the specific receptor for VAR2CSA, the oncofetal chondroitin sulfate (ofCS), is likewise present in BL tissue and cell lines. We therefore explored...

Development of melt compositions for sulphate bearing high level waste

International Nuclear Information System (INIS)

Jahagirdar, P.B.; Wattal, P.K.

1997-09-01

The report deals with the development and characterization of vitreous matrices for sulphate bearing high level waste. Studies were conducted in sodium borosilicate and lead borosilicate systems with the introduction of CaO, BaO, MgO etc. Lead borosilicate system was found to be compatible with sulphate bearing high level wastes. Detailed product evaluation carried on selected formulations is also described. (author)

A thermo-responsive and photo-polymerizable chondroitin sulfate-based hydrogel for 3D printing applications

NARCIS (Netherlands)

Abbadessa, A.; Blokzijl, M. M.; Mouser, V. H. M.; Marica, P.; Malda, J.; Hennink, W. E.; Vermonden, T.

2016-01-01

The aim ofthis study was to design a hydrogel system based on methacrylated chondroitin sulfate (CSMA) and a thermo-sensitive poly(N-(2-hydroxypropyl) methacrylamide-mono/dilactate)-polyethylene glycol triblock copolymer (M15P10) as a suitable material for additive manufacturing of scaffolds. CSMA

NUTRITIONAL VALUE AND HEAVY METALS CONTENTS OF THE DRIED SEA CUCUMBER Stichopus vastus FROM SALEMO ISLAND, INDONESIA

Directory of Open Access Journals (Sweden)

Abdullah Rasyid

2018-01-01

Full Text Available The dried sea cucumber Stichopus vastus is one of the commercially species harvested in Indonesian waters. This study aims to highlight the nutritional value and heavy metals content of dried sea cucumber S. vastus. Proximate (moisture, ash, protein, fat and carbohydrate, mineral (sodium, calcium, potassium and iron and heavy metal (mercury, cadmium, arsenic and lead were determined by standard method of AOAC, while phosphorous was determined by spectrophotometric method. Chondroitin sulphate was determined by UPLC method, glucosamine sulphate and vitamin (A, B1, B2 and E by HPLC method. Results show that protein was the major component in proximate analysis of dried sea cucumber S. vastus in the present study. The protein content was 38.70%. Moisture, ash, fat and carbohydrate content were 19.46%, 34.04%, 0.38% and 7.42% respectively. All vitamins and heavy metals examined in this study were not detected. The sodium content was 8054.36 mg/100 g higher than other minerals. Calcium, potassium, phosphorus and iron content were 2449.9 mg/100 g, 159.77 mg/100 g, 5085.2 mg/100 g and 520.8 mg/100 g respectively. Glucosamine sulphate content was found to be 2.429 g/100 g, whereas chondroitin sulphate was found to be 1.115 g/100 g. It can therefore, be concluded that the dried sea cucumber S. vastus from Salemo Island is safe for human consumption and hence can be used as a source of food supplement in the future. Keywords: food supplement, Salemo island, Stichopus vastus

[Sensitive Determination of Chondroitin Sulfate by Fluorescence Recovery of an Anionic Aluminum Phthalocyanine-Cationic Surfactant Ion-Association Complex Used as a Fluorescent Probe Emitting at Red Region].

Science.gov (United States)

Chen, Lin; Huang, Ping; Yang, Hui-qing; Deng, Ya-bin; Guo, Meng-lin; Li, Dong-hui

2015-08-01

Determination of chondroitin sulfate in the biomedical field has an important value. The conventional methods for the assay of chondroitin sulfate are still unsatisfactory in sensitivity, selectivity or simplicity. This work aimed at developing a novel method for sensitive and selective determination of chondroitin sulfate by fluorimetry. We found that some kinds of cationic surfactants have the ability to quench the fluorescence of tetrasulfonated aluminum phthalocyanine (AlS4Pc), a strongly fluorescent compound which emits at red region, with high efficiency. But, the fluorescence of the above-mentioned fluorescence quenching system recovered significantly when chondroitin sulfate (CS) exits. Tetradecyl dimethyl benzyl ammonium chloride(TDBAC) which was screened from all of the candidates of cationic surfactants was chosen as the quencher because it shows the most efficient quenching effect. It was found that the fluorescence of AlS4Pc was extremely quenched by TDBAC because of the formation of association complex between AlS4Pc and TDBAC. Fluorescence of the association complex recovered dramatically after the addition of chondroitin sulfate (CS) due to the ability of chondroitin sulfate to shift the association equilibrium of the association, leading to the release of AlS4Pc, thus resulting in an increase in the fluorescence of the reaction system. Based on this phenomenon, a novel method with simplicity, accuracy and sensitivity was developed for quantitative determination of CS. Factors including the reaction time, influencing factors and the effect of coexisting substances were investigated and discussed. Under optimum conditions the linear range of the calibration curve was 0.20~10.0 μg · mL(-1). The detection limit for CS was 0.070 μg · mL(-1). The method has been applied to the analysis of practical samples with satisfied results. This work expands the applications of AlS4Pc in biomedical area.

Pathophysiology of NG2-glia:a ‘Chicken and Egg’ scenario of altered neurotransmission and disruption of NG2-glial cell function

OpenAIRE

Rivera, Andrea Domenico; De La Rocha, Irene Chacon; Neville, Rebekah; Butt, Arthur Morgan

2016-01-01

Classically, the central nervous system (CNS) was considered to contain neurons and three main types of glial cells - astrocytes, oligodendrocytes, and microglia. Now, it has been clearly established that NG2-glia are a fourth glial cell type that are defined by their expression of the NG2 chondroitin sulfate proteoglycan (Cspg4). NG2-glia are also known as oligodendrocyte precursor cells (OPCs) and express the alpha receptor for platelet-derived growth factor (Pdgfra) as well as other oligod...

Coarctation induces alterations in basement membranes in the cardiovascular system

DEFF Research Database (Denmark)

Lipke, D W; McCarthy, K J; Elton, T S

1993-01-01

ventricular hypertrophy was maximal within 5 days. In immunohistochemical studies, fibronectin and laminin were increased and the basement membrane chondroitin sulfate proteoglycan decreased in both the subendothelial space and smooth muscle cell basement membranes of the aorta above the clip compared...... membrane components in the heart and vasculature peaked before maximal cardiac hypertrophy (5 days). These studies indicate that alterations in basement membrane component deposition in the hypertrophied vasculature occur at both transcriptional and translational levels and suggest that the cell attachment...

Differential inhibition of polymorphonuclear leukocyte recruitment in vivo by dextran sulphate and fucoidan

Directory of Open Access Journals (Sweden)

N. Van Osselaer

1996-01-01

Full Text Available The selectin-mediated rolling of leukocytes along the endothelial cells is a prerequisite step followed by firm adhesion and extravasation into the inflamed tissue. This initial contact can be suppressed by sulphated polysaccharides. We have studied the effect of sulphated polysaccharides on the ultimate polymorphonuclear leukocyte (PMN recruitment and plasma leakage in rabbit skin in response to intradermal injection of various inflammatory mediators. PMN infiltration evoked by various PMN chemoattractants (FMLP, C5a desArg, LTB4 and IL-8 was significantly inhibited after intravenous injection of dextran sulphate (25 mg/kg, heparin (2 × 90 mg/kg or fucoidan (1 mg/kg. PMN-dependent plasma leakage was equally well reduced by the different sulphated polymers. Vascular permeability induced by histamine or thrombin acting via a PMN-independent mechanism was not reduced. Fucoidan was the only polysaccharide able to suppress IL-1-induced PMN infiltration for 60–70%. Local administration of dextran sulphate had no effect on PMN-dependent plasma leakage. Differential inhibition of PMN recruitment was determined after injection of dextran sulphate or fucoidan depending on the type of insult. Therefore, these results suggest that different adhesion pathways are utilized during PMN recruitment in vivo in response to chemoattractants and IL-1.

The reducibility of sulphuric acid and sulphate in aqueous solution (translated from German)

International Nuclear Information System (INIS)

Grauer, R.

1990-07-01

In connection with the Swedish project for the final storage of spent fuel elements it was necessary to assess whether dissolved sulphate can corrode the copper canister without the intervention of sulphate-reducing bacteria. A simple reaction between copper and sulphate is thermodynamically impossible. On the other hand, copper can react to give copper sulphide if an additional electron donor such as iron is available. Because little specific information is available about this subject the problem was extended to the much more general question of the reducibility of sulphur in dilute aqueous solution. It is a part of the general knowledge of chemistry, and there is also unanimity about it in the geochemical literature, that purely chemical reduction of sulphate does not take place in dilute solution at temperatures below 100 degrees C. This fact is, however, poorly documented and it was therefore necessary to substantiate it by drawing on numerous individual findings from different areas of pure and applied chemistry. The investigation confirms that sulphur in dilute solution is completely inert towards chemical reducing agents and also to cathodic reduction. Thus corrosion of copper by sulphate under final-storage conditions and in the absence of sulphate reducing bacteria can be ruled out with a probability verging on certainty. (85 refs.)

DISTRIBUTION OF GBM HEPARAN-SULFATE PROTEOGLYCAN CORE PROTEIN AND SIDE-CHAINS IN HUMAN GLOMERULAR-DISEASES

NARCIS (Netherlands)

VANDENBORN, J; VANDENHEUVEL, LPWJ; BAKKER, MAH; VEERKAMP, JH; ASSMANN, KJM; WEENING, JJ; BERDEN, JHM

Using monoclonal antibodies (mAbs) recognizing either the core protein or the heparan sulfate (HS) side chain of human GBM heparan sulfate proteoglycan (HSPG), we investigated their glomerular distribution on cryostat sections of human kidney tissues. The study involved 95 biopsies comprising twelve

A thermo-responsive and photo-polymerizable chondroitin sulfate-based hydrogel for 3D printing applications

NARCIS (Netherlands)

Abbadessa, A|info:eu-repo/dai/nl/369480376; Blokzijl, M M; Mouser, V H M; Marica, P; Malda, J|info:eu-repo/dai/nl/412461099; Hennink, W E|info:eu-repo/dai/nl/070880409; Vermonden, T|info:eu-repo/dai/nl/275124517

2016-01-01

The aim of this study was to design a hydrogel system based on methacrylated chondroitin sulfate (CSMA) and a thermo-sensitive poly(N-(2-hydroxypropyl) methacrylamide-mono/dilactate)-polyethylene glycol triblock copolymer (M15P10) as a suitable material for additive manufacturing of scaffolds. CSMA

Influence on ammonium-sulphate fertilizer on P-uptake by upland rice

International Nuclear Information System (INIS)

Abdullah, N; Mitrosuhardjo, M.M.

1988-01-01

A greenhouse experiment to study the influence of ammonium -sulphate fertilizer on P-uptake by Atomita I/1 rice variety on latosol soil of Pasar Jumat has been conducted. Factorial experiment consisting of eight treatments was arrange in a fully randomized design. Each treatment was replicated three times. The rate of ammonium sulphate fertilizer was equivalent to 0, 30, 60, and 90 kg N/ha, and TSP at the rate of 90 and 180 kg P/ha. Ammonium sulphate fertilizer with 2% atom 15-N and TSP with specific activity of 0.2 mCi/g P2O5 were used in the experiment. The plant was harvested 70 days after planting, It was turned out that ammonium sulphate fertilizer gave a highly significant difference (p=<0.01) of dry weight of plant. The rate of TSP did not effect significantly on plant dry weight. Ammonium sulphate fertilizer gave highly significant difference on P-uptake. Fertilizer use efficiency was in the range of 3.88 - 6.60% for 90 kg P and 2.51 - 4.46% for 180 kg P. Fertilizer-N uptake was significantly increased (p=<0.05) with the increase of N rate. Rate of fertilizer-N use efficiency was in the range of 47-60% of the applied fertilizer. (authors). 4 refs, 9 tabs

Structural analysis of isomeric chondroitin sulfate oligosaccharides using regioselective 6-O-desulfation method and tandem mass spectrometry.

Science.gov (United States)

Chen, Shu-Ting; Her, Guor-Rong

2014-09-16

A strategy based on a regioselective 6-O-desulfation reaction and negative ion electrospray ionization tandem mass spectrometry (ESI-MS(n)) was developed for the structural delineation of isomeric chondroitin sulfate oligosaccharides. Product ions resulting from the glycosidic cleavage provided information about the number of sulfate groups in each sugar residue. After the regioselective 6-O-desulfation reaction, the number of sulfate groups on each residue was obtained using a tandem mass spectrometry analysis of the reaction product. The sulfation pattern could be obtained based on the product ions of analytes before and after the desulfation reaction. The strategy was demonstrated using a series of tetrasaccharides prepared from shark cartilage chondroitin sulfate D. Among the 12 identified tetrasaccharides, six structures had not been reported before. Copyright © 2014 Elsevier B.V. All rights reserved.

Novel single-chain antibody GD3A10 defines a chondroitin sulfate biomarker for ovarian cancer

NARCIS (Netherlands)

Vallen, M.J.E.; Tilborg, A.G. van; Tesselaar, M.H.; Dam, G.B. ten; Bulten, J.; Kuppevelt, T.H. van; Massuger, L.F.A.G.

2014-01-01

AIMS: Ovarian cancer has the highest case-to-fatality-index of all gynecological cancers. In this study, tumor-related alterations in the extracellular matrix, especially regarding chondroitin sulfate glycosaminoglycans, are proposed as a novel biomarker in ovarian cancer. MATERIALS & METHODS: Phage

Tyrosine sulphation is not required for microvillar expression of intestinal aminopeptidase N

DEFF Research Database (Denmark)

Danielsen, E M

1988-01-01

incorporation of [35S]sulphate into aminopeptidase N and other major microvillar hydrolases by 70-85% compared with controls, indicating an inhibition of their post-translational tyrosine sulphation. In labelling experiments with [35S]methionine from 0.5 to 5 h, DCNP was tested for its possible influence...... on synthesis, processing and microvillar expression of aminopeptidase N, but no effect on any of these parameters could be detected. It can therefore be concluded that tyrosine sulphation is not required (for instance as a sorting signal) for the targeting of newly synthesized enzymes to the microvillar...

Chondroitinase C Selectively Degrades Chondroitin Sulfate Glycosaminoglycans that Inhibit Axonal Growth within the Endoneurium of Peripheral Nerve.

Science.gov (United States)

Graham, James B; Muir, David

2016-01-01

The success of peripheral nerve regeneration is highly dependent on the regrowth of axons within the endoneurial basal lamina tubes that promote target-oriented pathfinding and appropriate reinnervation. Restoration of nerve continuity at this structural level after nerve transection injury by direct repair and nerve grafting remains a major surgical challenge. Recently, biological approaches that alter the balance of growth inhibitors and promoters in nerve have shown promise to improve appropriate axonal regeneration and recovery of peripheral nerve function. Chondroitin sulfate proteoglycans (CSPGs) are known inhibitors of axonal growth. This growth inhibition is mainly associated with a CSPG's glycosaminoglycan chains. Enzymatic degradation of these chains with chondroitinase eliminates this inhibitory activity and, when applied in vivo, can improve the outcome of nerve repair. To date, these encouraging findings were obtained with chondroitinase ABC (a pan-specific chondroitinase). The aim of this study was to examine the distribution of CSPG subtypes in rodent, rabbit, and human peripheral nerve and to test more selective biological enzymatic approaches to improve appropriate axonal growth within the endoneurium and minimize aberrant growth. Here we provide evidence that the endoneurium, but not the surrounding epineurium, is rich in CSPGs that have glycosaminoglycan chains readily degraded by chondroitinase C. Biochemical studies indicate that chondroitinase C has degradation specificity for 6-sulfated glycosaminoglycans found in peripheral nerve. We found that chondroitinase C degrades and inactivates inhibitory CSPGs within the endoneurium but not so much in the surrounding nerve compartments. Cryoculture bioassays (neurons grown on tissue sections) show that chondroitinase C selectively and significantly enhanced neuritic growth associated with the endoneurial basal laminae without changing growth-inhibiting properties of the surrounding epineurium

Oxygen isotopic composition of sulphates from some mineral waters and mine waters in western Bohemia

International Nuclear Information System (INIS)

Smejkal, V.

1979-01-01

Two main genetic types of sulphate can be distinguished according to the delta 18 O(SO 4 2- ) measurements in sulphate-rich mineral and mine waters of western Bohemia - sulphates in descending mine waters and in weathered outcrops of graphitic pyrite slates from areas outside Tertiary basins, which originated in recent time by the oxidation of sulphides in the presence of atmospheric oxygen, have delta 18 O values from -2.1 to -6.1 per mille SMOW; and sulphates from springs of mineral waters of the renowned spas of Karlovy Vary, Frantiskovy Lazne and Marianske Lazne show distinctly heavier delta 18 O values - from +4.0 to +6.4 per mille, with maximum of values between +5.0 and +6.0 per mille. Similar delta 18 O values have been established in thenardites and in gypsum in Miocene claystones and in sulphates of some mine waters in the nearby Tertiary Cheb and Sokolov Basins. The presented results indicate that sulphates in mineral waters of the Karlovy Vary type originate mainly by leaching of Miocene sulphates and not by present-day oxidation of sulphidic sulphur. (author)

Possibilities of Sulphate Elimination from Mine Water

Directory of Open Access Journals (Sweden)

Heviánková Silvie

2004-12-01

Full Text Available The issue of „acid water“ (or AMD is well known in the world for some centuries. In the Eastern Slovakia, the most acid surface water occurs in the area of old mine Smolník, which is closed and submerged for 15 years. The submitted contribution deals with the sulphate-elimination from this locality. Recently, several methods of sulphate-elimination from the mine water are applied. The best-known methods are biological, physical-chemical and chemical precipitation. The method described in this contribution consists of chemical precipitation by sodium aluminate and calcium hydrate. Under application of this method very interesting results were obtained. The amount of SO42- anions decreased to almost zero-value, using optimal doses of the chemical reagents.

Phosphatidylcholine nanovesicles coated with chitosan or chondroitin sulfate as novel devices for bacteriocin delivery

Science.gov (United States)

da Silva, Indjara Mallmann; Boelter, Juliana Ferreira; da Silveira, Nádya Pesce; Brandelli, Adriano

2014-07-01

There is increased interest on the use of natural antimicrobial peptides in biomedicine and food preservation technologies. In this study, the antimicrobial activity of nisin encapsulated into nanovesicles containing polyanionic polysaccharides was investigated. Nisin was encapsulated in phosphatidylcholine (PC) liposomes containing chitosan or chondroitin sulfate by the thin-film hydration method and tested for antimicrobial activity against Listeria spp. The mean particle size of PC liposomes was 145 nm and varied to 210 and 134 nm with the incorporation of chitosan and chondroitin sulfate, respectively. Nisin-containing nanovesicles with and without incorporation of polysaccharides had a zeta potential values around -20 mV, showing mostly spherical structures when observed by transmission electron microscopy. Encapsulated nisin had similar efficiency as free nisin in inhibiting Listeria spp. isolated from bovine carcass, and greater efficiency in inhibiting Listeria monocytogenes. The formulation containing chitosan was more stable and more efficient in inhibiting L. monocytogenes when compared to the other nanovesicles tested. After 24 h, the viable cell counts were 2 log lower as compared with the other treatments and 7 log comparing to controls.

The Metal And Sulphate Removal From Mine Drainage Waters By Biological-Chemical Ways

Directory of Open Access Journals (Sweden)

Jenčárová Jana

2015-06-01

Full Text Available Mine drainage waters are often characterized by high concentrations of sulphates and metals as a consequence of the mining industry of sulphide minerals. The aims of this work are to prove some biological-chemical processes utilization for the mine drainage water treatment. The studied principles of contamination elimination from these waters include sulphate reduction and metal bioprecipitation by the application of sulphate-reducing bacteria (SRB. Other studied process was metal sorption by prepared biogenic sorbent. Mine drainage waters from Slovak localities Banská Štiavnica and Smolník were used to the pollution removal examination. In Banská Štiavnica water, sulphates decreased below the legislative limit. The elimination of zinc by sorption experiments achieved 84 % and 65 %, respectively.

An Injectable Enzymatically Crosslinked Carboxymethylated Pullulan/Chondroitin Sulfate Hydrogel for Cartilage Tissue Engineering

Science.gov (United States)

Chen, Feng; Yu, Songrui; Liu, Bing; Ni, Yunzhou; Yu, Chunyang; Su, Yue; Zhu, Xinyuan; Yu, Xiaowei; Zhou, Yongfeng; Yan, Deyue

2016-01-01

In this study, an enzymatically cross-linked injectable and biodegradable hydrogel system comprising carboxymethyl pullulan-tyramine (CMP-TA) and chondroitin sulfate-tyramine (CS-TA) conjugates was successfully developed under physiological conditions in the presence of both horseradish peroxidase (HRP) and hydrogen peroxide (H2O2) for cartilage tissue engineering (CTTE). The HRP crosslinking method makes this injectable system feasible, minimally invasive and easily translatable for regenerative medicine applications. The physicochemical properties of the mechanically stable hydrogel system can be modulated by varying the weight ratio and concentration of polymer as well as the concentrations of crosslinking reagents. Additionally, the cellular behaviour of porcine auricular chondrocytes encapsulated into CMP-TA/CS-TA hydrogels demonstrates that the hydrogel system has a good cyto-compatibility. Specifically, compared to the CMP-TA hydrogel, these CMP-TA/CS-TA composite hydrogels have enhanced cell proliferation and increased cartilaginous ECM deposition, which significantly facilitate chondrogenesis. Furthermore, histological analysis indicates that the hydrogel system exhibits acceptable tissue compatibility by using a mouse subcutaneous implantation model. Overall, the novel injectable pullulan/chondroitin sulfate composite hydrogels presented here are expected to be useful biomaterial scaffold for regenerating cartilage tissue.

Volcanic lake systems as terrestrial analogue for sulphate-rich terrains on Mars

NARCIS (Netherlands)

Rodríguez, A.

2016-01-01

Remote-sensing observations and rover missions have documented the abundant presence of sulphate-rich mineral associations on the surface of Mars. Together with widespread occurrences of silica and frequent enrichments of chlorine and bromine in soils and rocks, the sulphate associations are

Altered Liver Proteoglycan/Glycosaminoglycan Structure as a Manifestation of Extracellular Matrix Remodeling upon BCG-induced Granulomatosis in Mice.

Science.gov (United States)

Kim, L B; Shkurupy, V A; Putyatina, A N

2017-01-01

Experimental BCG-induced granulomatosis in mice was used to study changes in the dynamics of individual liver proteoglycan components reflecting phasic extracellular matrix remodeling, determined by the host-parasite interaction and associated with granuloma development. In the early BCG-granulomatosis period, the increase in individual proteoglycan components promotes granuloma formation, providing conditions for mycobacteria adhesion to host cells, migration of phagocytic cells from circulation, and cell-cell interaction leading to granuloma development and fibrosis. Later, reduced reserve capacity of the extracellular matrix, development of interstitial fibrosis and granuloma fibrosis can lead to trophic shortage for cells within the granulomas, migration of macrophages out of them, and development of spontaneous necrosis and apoptosis typical of tuberculosis.

Risk minimisation of FGD gypsum leachates by incorporation of aluminium sulphate

Energy Technology Data Exchange (ETDEWEB)

Alvarez-Ayuso, E. [Department of Environmental Geology, Institute of Earth Sciences ' Jaume Almera' (CSIC), C/ Lluis Sole i Sabaris, s/n, 08028 Barcelona (Spain); Department of Environmental Geochemistry, IRNASA, CSIC, Apto. 257, 37071 Salamanca (Spain)], E-mail: ealvarez@ija.csic.es; Querol, X. [Department of Environmental Geology, Institute of Earth Sciences ' Jaume Almera' (CSIC), C/ Lluis Sole i Sabaris, s/n, 08028 Barcelona (Spain); Ballesteros, J.C.; Gimenez, A. [Endesa Generacion, S.A., C/ Ribera de Loira, 60, 28042 Madrid (Spain)

2008-11-15

The incorporation of aluminium sulphate to (flue gas desulphurisation) FGD gypsum before its disposal was investigated as a way to minimise the risk supposed by the high fluoride content of its leachates. Using a bath method the kinetic and equilibrium processes of fluoride removal by aluminium sulphate were studied at fluoride/aluminium molar concentration (F/Al) ratios in the range 1.75 10{sup -2}-1.75 under the pH conditions (about 6.5) of FGD gypsum leachates. It was found that fluoride removal was a very fast process at any of the (F/Al) ratios subject of study, with equilibrium attained within the first 15 min of interaction. High decreases in solution fluoride concentrations (50-80%) were found at the equilibrium state. The use of aluminium sulphate in the stabilization of FGD gypsum proved to greatly decrease its fluoride leachable content (in the range 20-90% for aluminium sulphate doses of 0.1-5%, as determined by the European standard EN 12457-4). Such fluoride leaching minimisation assures the characterization of this by-product as a waste acceptable at landfills for non-hazardous wastes according to the Council Decision 2003/33/EC on waste disposal. Furthermore, as derived from column leaching studies, the proposed stabilization system showed to be highly effective in simulated conditions of disposal, displaying fluoride leaching reduction values about 55 and 80% for aluminium sulphate added amounts of 1 and 2%, respectively.

Perlecan (basement membrane heparan sulfate proteoglycan and its role in oral malignancies: An overview

Directory of Open Access Journals (Sweden)

Mithilesh Mishra

2011-01-01

Full Text Available Perlecan means pearl-like structures. Perlecan is a large proteoglycan (400-500 kDa present in virtually all vascularized tissues with a distribution that is primarily confined to basement membranes including those of oral mucosa. It is a basement membrane-type heparan sulfate proteoglycan. Perlecan is synthesized by basal cells and fibroblasts adjacent to the basal lamina . Perlecan is also synthesized by vascular endothelial and smooth muscle cells present in the extracellular matrix. It has been demonstrated in recent years that perlecan is distributed in the stromal space of various pathophysiological conditions. The complex pleiotropy of perlecan suggests that this gene product is involved in several developmental processes, at both early and late stages of embryogenesis, as well as in cancer and diabetes. In the oral cavity, perlecan expression is reported to basal cells in normal mucosa and its expression increases in precancer and cancerous conditions. It is also expressed in various odontogenic tumors such as ameloblastoma, keratocyst odontogenic tumor, and also salivary gland tumors such as adenoid cystic carcinoma, mucoepidermoid carcinoma, etc.

Abiotic nitrate and sulphate reduction by hydrogen: a comparative experimental study

International Nuclear Information System (INIS)

Truche, L.; Berger, G.; Albrecht, A.; Giffaut, E.

2010-01-01

Document available in extended abstract form only. The bituminous waste which is part of the intermediate level, long-lived waste (MAVL) is characterised, amongst others, by the coexistence of nitrates, sulphates, organic matter, native metals and hydrogen gas in the waste mixture and package. It can be considered as the most complex example that will be used to discuss redox reactions occurring in such waste mixtures. The evaluation of the redox conditions requires quantification of the amount of electron acceptors and donors and definition of the kinetics of redox reaction. The objectives of an experimental study to unravel some of these reaction complexities are: - to investigate nature and rate of sulphate and nitrate reduction by hydrogen in the presence of different catalysts (stainless steel, hastelloy, magnetite and argillite); - to compare sulphate and nitrate as electron acceptors; - to provide a mechanistic model of these reactions. It is well known that reduction of sulphate and nitrate requires high activation energies, usually supplied either by thermal processes or via bacterial and surface catalysis, of which the latter has been investigated in this study. Preliminary experiments performed at 150 deg. C and under H 2 pressure show that sulphate reduction is enhanced in the presence of magnetite, but essentially under the restricted condition of low sulphate concentration and at a pH below the Point of Zero Charge of magnetite. This suggests that sorption of sulphate contributes to the catalysed reaction (at low pH) but provided that the magnetite surface sites are not saturated with respect to aqueous sulphate (low concentration). On the contrary, nitrate reduction is observed whatever the pH and the nitrate concentration in the presence of both magnetite and hastelloy C276 (Ni, Cr, Mo, W, Fe alloy). The effect of temperature on the rate of nitrate reduction (500 ppm KNO 3 solution) is shown by comparing three different experiments conducted in

Sulphate content of the Muntimpa dam water and its impact on water quality

International Nuclear Information System (INIS)

Tembo, F; Shitumbanuma, V; Simukanga, S; Mudenda, G; Chileshe, P; Mulenga, S; Phiri, Y

2004-01-01

This article presents results of a study of the quality of water from Muntimpa Dam, a reservior of waste mine water released from the processing of copper and cobalt ores by Konkola Copper Mines(KCM) Plc in Chingola. The mine water is discharged into the local Muntimpa stream, a possible source of drinking and domestic water for the local population. The purpose of the study was to determine levels of sulphate in the dam and stream water and recommend possible methods of partial sulphate removal to levels below the recommended statutory limits and secondly, to assess the impact of high sulphate levels on water quality. Study methods included the sampling of water from the Muntimpa dam and catchment area. Stream water samples were collected about 5m from the stream banks while water samples from the dam were randomly collected from the near the centre of the dam at a depth of 50cm. Laboratory methods involved the determination of physical and chemical properties of the water using standard analytical techniques. Results of the study indicate that both total (2470mg/l) and available (1965mg/l) sulphate concentrations are higher than the recommended statutory limit for the discharge of sulphates into natural streams of 1500mg/l. From the study it is concluded that water in Muntimpa dam and stream is not suitable for drinking and other domestic use due to the high sulphate levels. From theorectical considerations, it was established that sulphate reduction could be achieved by addition of lime, which however had the consquence of increasing the pH of the water in excess of the recommended Zambian statutory value of nine, and would thus require an additional process to reduce the pH. (author)

Grass-cellulose as energy source for biological sulphate removal from acid mine effluents

CSIR Research Space (South Africa)

Greben, HA

2008-11-01

Full Text Available The biological sulphate removal technology requires carbon and energy sources to reduce sulphate to sulphide. Plant biomass, e.g. grass, is a sustainable source of energy when cellulose is utilised during anaerobic degradation, producing volatile...

Preparation and antifouling property of polyurethane film modified by chondroitin sulfate

Science.gov (United States)

Yuan, Huihui; Xue, Jing; Qian, Bin; Chen, Huaying; Zhu, Yonggang; Lan, Minbo

2017-02-01

An antifouling polyurethane film modified by chondroitin sulfate (PU-CS) was prepared by chemical grafting with N-Boc-1,3-propanediamine as a spacer. The different mass fraction of N-Boc-1,3-propanediamine was investigated to obtain PU-CS films with different CS grafting density. The surface properties of PU-CS films were comprehensively characterized. Proteins adsorption and glycosaminoglycans adhesion on films were evaluated. Moreover, inorganic salt deposition on film with highest CS grafting density (3.70 μg/cm2) was briefly investigated. The results showed that the increase of CS grafting density improved not only the hydrophilicity but the antifouling performance of films. The best antifouling film reduced the adsorption of fibrinogen (BFG), human serum albumin (HSA) and lysozyme (LYS) by 81.4%, 95.0% and 76.5%, respectively, and the adhesion of chondroitin (CS), heparin (HP) and hyaluronic acid (HA) by 70.6%, 87.4% and 81.3%, respectively. In addition, the co-adsorption of proteins and glycosaminoglycans reduced up to 86.9% and 75.5%, respectively. Changes in inorganic salt deposition after co-adsorption of proteins and glycosaminoglycans on PU-CS(3) suggested that the proteins promoted the inorganic salt deposition, while glycosaminoglycans inhibited the crystal growth. The negatively charged polysaccharides might promote the generation of smaller crystals which could be conducive to provide theoretical and practical guide to develop novel urinary stents with significant anti-encrustation properties.

Development of a glass matrix for vitrification of sulphate bearing high level radioactive liquid waste

International Nuclear Information System (INIS)

Kaushik, C.P.; Mishra, R.K.; Thorat, Vidya; Ramchandran, M.; Amar Kumar; Ozarde, P.D.; Raj, Kanwar; Das, D.

2004-07-01

High level radioactive liquid waste (HLW) is generated during reprocessing of spent nuclear fuel. In the earlier reprocessing flow sheet ferrous sulphamate has been used for valancy adjustment of Pu from IV to III for effective separation. This has resulted in generation of HLW containing significance amount of sulphate. Internationally borosilicate glass matrix has been adopted for vitrification of HLW. The first Indian vitrification facility at Waste Immobilislition Plant (WIP), Tarapur a five component borosilicate matrix (SiO 2 :B 2 O 3 :Na 2 O : MnO : TiO 2 ) has been used for vitrification of waste. However at Trombay HLW contain significant amount of sulphate which is not compatible with standard borosilicate formulation. Extensive R and D efforts were made to develop a glass formulation which can accommodate sulphate and other constituents of HLW e.g., U, Al, Ca, etc. This report deals with development work of a glass formulations for immobilization of sulphate bearing waste. Different glass formulations were studied to evaluate the compatibility with respect to sulphate and other constituents as mentioned above. This includes sodium, lead and barium borosilicate glass matrices. Problems encountered in different glass matrices for containment of sulphate have also been addressed. A glass formulation based on barium borosilicate was found to be effective and compatible for sulphate bearing high level waste. (author)

The chondroitin sulfate A-binding site of the VAR2CSA protein involves multiple N-terminal domains

DEFF Research Database (Denmark)

Dahlbäck, Madeleine; Jørgensen, Lars M; Nielsen, Morten A

2011-01-01

Malaria during pregnancy is a major health problem for African women. The disease is caused by Plasmodium falciparum malaria parasites, which accumulate in the placenta by adhering to chondroitin sulfate A (CSA). The interaction between infected erythrocytes and the placental receptor is mediated...

Molecular alterations of tropoelastin and proteoglycans induced by tobacco smoke extracts and ultraviolet A in cultured skin fibroblasts

International Nuclear Information System (INIS)

Yin, Lei; Morita, Akimichi; Tsuji, Takuo

2002-01-01

Functional integrity of normal skin is dependent on the balance between the biosynthesis and degradation of extracellular matrix, primarily composed of collagen, elastin and proteoglycans. In our previous studies, we found that tobacco smoke extracts decreased expressions of type I and III procollagen and induced matrix metalloproteinase-1 (MMP-1) and MMP-3 in the cultured skin fibroblasts. We here further investigated the effects of tobacco smoke extracts or ultraviolet A (UVA) treatments on the expression of tropoelastin (soluble elastin protein), and versican and decorin (proteoglycans) in cultured skin fibroblasts. The mRNA of tropoelastin increased by tobacco smoke extracts or UVA irradiation. Versican was markedly shown to decrease after these treatments by using western blotting and the mRNA of versican V0 also significantly decreased. UVA treatment did not show remarkable change in decorin protein, but resulted in marked decrease of decorin D1 mRNA. In contrast to UVA irradiation, the treatments of tobacco smoke extracts resulted in significant increase in decorin, while mRNA of decorin D1 decreased as compared to the control. MMP-7 increased after the treatment of tobacco smoke extracts or UVA. These results indicated that common molecular features might underlie the skin premature aging induced by tobacco smoke extracts and UVA, including abnormal regulation of extracellular matrix deposition through elevated MMPs, reduced collagen production, abnormal tropoelastin accumulation, and altered proteoglycans. (author)

Molecular alterations of tropoelastin and proteoglycans induced by tobacco smoke extracts and ultraviolet A in cultured skin fibroblasts

Energy Technology Data Exchange (ETDEWEB)

Yin, Lei; Morita, Akimichi; Tsuji, Takuo [Nagoya City Univ. (Japan). Medical School

2002-02-01

Functional integrity of normal skin is dependent on the balance between the biosynthesis and degradation of extracellular matrix, primarily composed of collagen, elastin and proteoglycans. In our previous studies, we found that tobacco smoke extracts decreased expressions of type I and III procollagen and induced matrix metalloproteinase-1 (MMP-1) and MMP-3 in the cultured skin fibroblasts. We here further investigated the effects of tobacco smoke extracts or ultraviolet A (UVA) treatments on the expression of tropoelastin (soluble elastin protein), and versican and decorin (proteoglycans) in cultured skin fibroblasts. The mRNA of tropoelastin increased by tobacco smoke extracts or UVA irradiation. Versican was markedly shown to decrease after these treatments by using western blotting and the mRNA of versican V0 also significantly decreased. UVA treatment did not show remarkable change in decorin protein, but resulted in marked decrease of decorin D1 mRNA. In contrast to UVA irradiation, the treatments of tobacco smoke extracts resulted in significant increase in decorin, while mRNA of decorin D1 decreased as compared to the control. MMP-7 increased after the treatment of tobacco smoke extracts or UVA. These results indicated that common molecular features might underlie the skin premature aging induced by tobacco smoke extracts and UVA, including abnormal regulation of extracellular matrix deposition through elevated MMPs, reduced collagen production, abnormal tropoelastin accumulation, and altered proteoglycans. (author)

Treatment of acid and sulphate-rich effluents in an integrated biological/chemical process

CSIR Research Space (South Africa)

Maree, JP

2004-04-01

Full Text Available .4 g SO4/(l.d). The rate of biological sulphate removal was found to be directly related to the square root of sulphate, COD and VSS concentrations respectively, and inversely proportional to sulphide concentration. The practical value of simultaneous...

Comparison of the effects of the K(+)-channel openers cromakalim and minoxidil sulphate on vascular smooth muscle.

Science.gov (United States)

Wickenden, A. D.; Grimwood, S.; Grant, T. L.; Todd, M. H.

1991-01-01

1 The actions of the potassium channel openers, cromakalim and minoxidil sulphate, were compared in a range of isolated blood vessel preparations. 2 Cromakalim and minoxidil sulphate inhibited spontaneous mechanical activity of the guinea-pig portal vein and relaxed the noradrenaline precontracted rat aorta with similar potency. In contrast, minoxidil sulphate was less potent than cromakalim in inhibiting spontaneous activity in the rat portal vein and was essentially inactive in the noradrenaline precontracted rat mesenteric artery and rabbit aorta. 3 Minoxidil sulphate did not antagonize the effects of cromakalim in the rabbit aorta indicating it was not acting as a partial 'agonist'. 4 Charybdotoxin, noxiustoxin and rubidium failed to discriminate between cromakalim and minoxidil sulphate indicating that the apparently selective effects of minoxidil sulphate were not mediated by either Ca(2+)-activated potassium channels, delayed rectifiers or rubidium impermeable potassium channels. 5 Glibenclamide antagonized the effects of cromakalim in an apparently competitive manner whereas the effects of minoxidil sulphate were antagonized in a non-competitive manner. The involvement of subtypes of ATP-sensitive potassium channels is discussed. PMID:1878752

Sodium Sulphate Effect on Cement Produced with Building Stone Waste

Directory of Open Access Journals (Sweden)

Emre Sancak

2015-01-01

Full Text Available In this study, the blended cements produced by using the building stone waste were exposed to sulphate solution and the cement properties were examined. Prepared mortar specimens were cured under water for 28 days and then they were exposed to three different proportions of sodium sulphate solution for 125 days. Performances of cements were determined by means of compressive strength and tensile strength tests. The broken parts of some mortar bars were examined with scanning electron microscope (SEM. Besides, they were left under moist atmosphere and their length change was measured and continuously monitored for period of 125 days. In blended cements, solely cements obtained by replacing 10–20% of diatomites gave similar strength values with ordinary Portland cement (CEM I 42.5R at the ages of 7, 28, and 56 days. In all mortar specimens that included either waste andesite (AP or marble powder (MP showed best performance against very severe effective sodium sulphate solutions (13500 mg/L.

Should Magnesium Sulphate Prophylaxis be Used in all Cases of Severe Preeclampsia?

Directory of Open Access Journals (Sweden)

Sohini Bhattacharya,

2011-01-01

Full Text Available A case control study, undertaken in the department of Obstetrics and Gynecology in a tertiary referral centre, was designed to administer standard dose of magnesium sulphate to 50 randomly selected severely preeclamptic women in labor (Group 1. 48 women who formed the control group did not receive the drug (Group 2. The admission – delivery interval, rate of Cesarean section showed no statistically significant difference. Four patients developed convulsions in Group 1 in contrast to twelve patients in Group 2. Efficacy of magnesium sulphate at preventing eclampsia was calculated as 68%. Mild respiratory depression occurred in two cases and oliguria in eight cases in Group 1. Both the conditions improved on suspending further doses of magnesium sulphate. No patient had post partum haemorrhage in Group 1 although 4% patients had it in Group 2. Neonatal outcome was comparable in both the groups. Hence magnesium sulphate may be regarded as a fairly safe and effective prophylactic agent for eclampsia when used in severe preeclampsia in labor.

Thermal decomposition of hydrotalcite with chromate, molybdate or sulphate in the interlayer

Energy Technology Data Exchange (ETDEWEB)

Frost, Ray L. [Inorganic Materials Research Program, School of Physical and Chemical Sciences, Queensland University of Technology, GPO Box 2434, Brisbane Queensland 4001 (Australia)]. E-mail: r.frost@qut.edu.au; Musumeci, Anthony W. [Inorganic Materials Research Program, School of Physical and Chemical Sciences, Queensland University of Technology, GPO Box 2434, Brisbane Queensland 4001 (Australia); Bostrom, Thor [Inorganic Materials Research Program, School of Physical and Chemical Sciences, Queensland University of Technology, GPO Box 2434, Brisbane Queensland 4001 (Australia); Adebajo, Moses O. [Inorganic Materials Research Program, School of Physical and Chemical Sciences, Queensland University of Technology, GPO Box 2434, Brisbane Queensland 4001 (Australia); Weier, Matt L. [Inorganic Materials Research Program, School of Physical and Chemical Sciences, Queensland University of Technology, GPO Box 2434, Brisbane Queensland 4001 (Australia); Martens, Wayde [Inorganic Materials Research Program, School of Physical and Chemical Sciences, Queensland University of Technology, GPO Box 2434, Brisbane Queensland 4001 (Australia)

2005-05-15

The thermal decomposition of hydrotalcites with chromate, molybdate and sulphate in the interlayer has been studied using thermogravimetric analysis coupled to a mass spectrometer measuring the gas evolution. X-ray diffraction shows the hydrotalcites have a d(0 0 3) spacing of 7.98 A with very small differences in the d-spacing between the three hydrotalcites. XRD was also used to determine the products of the thermal decomposition. For the sulphate-hydrotalcite decomposition the products were MgO and a spinel MgAl{sub 2}O{sub 4}, for the chromate interlayered hydrotalcite MgO, Cr{sub 2}O{sub 3} and spinel. For the molybdate interlayered hydrotalcite the products were MgO, spinel and MgMoO{sub 4}. EDX analyses enabled the formula of the hydrotalcites to be determined. Two processes are observed in the thermal decomposition namely dehydration and dehydroxylation and for the case of the sulphate interlayered hydrotalcite, a third process is the loss of sulphate. Both the dehydration and dehydroxylation take place in three steps each for each of the hydrotalcites.

Sustained Delivery of Chondroitinase ABC from Hydrogel System

Directory of Open Access Journals (Sweden)

Filippo Rossi

2012-03-01

Full Text Available In the injured spinal cord, chondroitin sulfate proteoglycans (CSPGs are the principal responsible of axon growth inhibition and they contribute to regenerative failure, promoting glial scar formation. Chondroitinase ABC (chABC is known for being able to digest proteoglycans, thus degrading glial scar and favoring axonal regrowth. However, its classic administration is invasive, infection-prone and clinically problematic. An agarose-carbomer (AC1 hydrogel, already used in SCI repair strategies, was here investigated as a delivery system capable of an effective chABC administration: the material ability to include chABC within its pores and the possibility to be injected into the target tissue were firstly proved. Subsequently, release kinetic and the maintenance of enzymatic activity were positively assessed: AC1 hydrogel was thus confirmed to be a feasible tool for chABC delivery and a promising device for spinal cord injury topic repair strategies.

Small leucine-rich repeat proteoglycans associated with mature insoluble elastin serve as binding sites for galectins.

Science.gov (United States)

Itoh, Aiko; Nonaka, Yasuhiro; Ogawa, Takashi; Nakamura, Takanori; Nishi, Nozomu

2017-11-01

We previously reported that galectin-9 (Gal-9), an immunomodulatory animal lectin, could bind to insoluble collagen preparations and exerted direct cytocidal effects on immune cells. In the present study, we found that mature insoluble elastin is capable of binding Gal-9 and other members of the human galectin family. Lectin blot analysis of a series of commercial water-soluble elastin preparations, PES-(A) ~ PES-(E), revealed that only PES-(E) contained substances recognized by Gal-9. Gal-9-interacting substances in PES-(E) were affinity-purified, digested with trypsin and then analyzed by reversed-phase HPLC. Peptide fragments derived from five members of the small leucine-rich repeat proteoglycan family, versican, lumican, osteoglycin/mimecan, prolargin, and fibromodulin, were identified by N-terminal amino acid sequence analysis. The results indicate that Gal-9 and possibly other galectins recognize glycans attached to small leucine-rich repeat proteoglycans associated with insoluble elastin and also indicate the possibility that mature insoluble elastin serves as an extracellular reservoir for galectins.

EFSA NDA Panel (EFSA Panel on Dietetic Products, Nutrition and Allergies), 2013 . Scientific Opinion on Rooster Combs Extract

DEFF Research Database (Denmark)

Tetens, Inge; Poulsen, Morten

, chondroitin sulphate A and dermatan sulphate. The applicant intends to add RCE to a number of dairy products with a recommended maximum intake of 80 mg RCE per portion and per day. The target population is the general population, with the exception of pregnant women, children and people with adverse reactions...... to sodium hyaluronate and/or avian protein. In the high intake scenario for “consumers only”, the highest daily intake would occur in adults in Belgium (0.788 g). The highest intake scenario for “all subjects” was estimated for adolescents in Denmark (0.427 g/day). The Panel notes that no adverse effects...

Sulfation and cation effects on the conformational properties of the glycan backbone of chondroitin sulfate disaccharides.

Science.gov (United States)

Faller, Christina E; Guvench, Olgun

2015-05-21

Chondroitin sulfate (CS) is one of several glycosaminoglycans that are major components of proteoglycans. A linear polymer consisting of repeats of the disaccharide -4GlcAβ1-3GalNAcβ1-, CS undergoes differential sulfation resulting in five unique sulfation patterns. Because of the dimer repeat, the CS glycosidic "backbone" has two distinct sets of conformational degrees of freedom defined by pairs of dihedral angles: (ϕ1, ψ1) about the β1-3 glycosidic linkage and (ϕ2, ψ2) about the β1-4 glycosidic linkage. Differential sulfation and the possibility of cation binding, combined with the conformational flexibility and biological diversity of CS, complicate experimental efforts to understand CS three-dimensional structures at atomic resolution. Therefore, all-atom explicit-solvent molecular dynamics simulations with Adaptive Biasing Force sampling of the CS backbone were applied to obtain high-resolution, high-precision free energies of CS disaccharides as a function of all possible backbone geometries. All 10 disaccharides (β1-3 vs β1-4 linkage × five different sulfation patterns) were studied; additionally, ion effects were investigated by considering each disaccharide in the presence of either neutralizing sodium or calcium cations. GlcAβ1-3GalNAc disaccharides have a single, broad, thermodynamically important free-energy minimum, whereas GalNAcβ1-4GlcA disaccharides have two such minima. Calcium cations but not sodium cations bind to the disaccharides, and binding is primarily to the GlcA -COO(-) moiety as opposed to sulfate groups. This binding alters the glycan backbone thermodynamics in instances where a calcium cation bound to -COO(-) can act to bridge and stabilize an interaction with an adjacent sulfate group, whereas, in the absence of this cation, the proximity of a sulfate group to -COO(-) results in two like charges being both desolvated and placed adjacent to each other and is found to be destabilizing. In addition to providing information

Proteoglycan Aggrecan Conducting T Cell Activation and Apoptosis in a Murine Model of Rheumatoid Arthritis

Directory of Open Access Journals (Sweden)

A. Hanyecz

2014-01-01

Full Text Available Rheumatoid arthritis (RA is a systemic autoimmune disease and its targeting of the joints indicates the presence of a candidate autoantigen(s in synovial joints. Patients with RA show immune responses in their peripheral blood to proteoglycan (PG aggrecan. One of the most relevant animal models of RA appears to be proteoglycan-induced arthritis (PGIA, and CD4+ T cells seem to play a crucial role in the initiation of the disease. In this review, the role of various T cell epitopes of aggrecan in the induction of autoreactive T cell activation and arthritis is discussed. We pay special attention to two critically important arthritogenic epitopes, 5/4E8 and P135H, found in the G1 and G3 domains of PG aggrecan, respectively, in the induction of autoimmune arthritis. Finally, results obtained with the recently developed PG-specific TCR transgenic mice system showed that altered T cell apoptosis, the balance of activation, and apoptosis of autoreactive T cells are critical factors in the development of autoimmunity.

Molecular structure of dextran sulphate sodium in aqueous environment

Science.gov (United States)

Yu, Miao; Every, Hayley A.; Jiskoot, Wim; Witkamp, Geert-Jan; Buijs, Wim

2018-03-01

Here we propose a 3D-molecular structural model for dextran sulphate sodium (DSS) in a neutral aqueous environment based on the results of a molecular modelling study. The DSS structure is dominated by the stereochemistry of the 1,6-linked α-glucose units and the presence of two sulphate groups on each α-glucose unit. The structure of DSS can be best described as a helix with various patterns of di-sulphate substitution on the glucose rings. The presence of a side chain does not alter the 3D-structure of the linear main chain much, but affects the overall spatial dimension of the polymer. The simulated polymers have a diameter similar to or in some cases even larger than model α-hemolysin nano-pores for macromolecule transport in many biological processes, indicating a size-limited translocation through such pores. All results of the molecular modelling study are in line with previously reported experimental data. This study establishes the three-dimensional structure of DSS and summarizes the spatial dimension of the polymer, serving as the basis for a better understanding on the molecular level of DSS-involved electrostatic interaction processes with biological components like proteins and cell pores.

Magnesium sulphate as an anticonvulsant in the management of eclampsia

International Nuclear Information System (INIS)

Ahmed, R.

2004-01-01

Objective: To evaluate the use of magnesium sulphate (MgSO/sub 4/) as an anticonvulsant in the management of eclamptic patients. Patients and Methods: Out of a total of 6050 pregnant women, 31 patients had eclampsia and were managed according to the set protocol. MgSO/sub 4/ 'Nas given to eclamptic patients according to protocol who had no contraindication to this agent. Recurrence of convulsion, side effects of magnesium sulphate, maternal and fetal outcome was noted. Results: Over two years' study period, out of 6050 patients, 31 were admitted with eclampsia (0.51 %). There was no maternal death. Out of 31 eclamptic patients only 5 patients were booked. Twenty-two patients (70.9%) were primigravida and 9 (29%) were multigravida. Seventeen (54%) were less than 20 years of age, 22 (79.9%) patients were admitted with antepartum and 6 (19.35%) had postpartum eclampsia. Magnesium sulphate was effective in 29 (93.54%) patients. Fifteen patients delivered by caesarean (C) section and 16 delivered vaginally. Twenty-nine (93.5%) babies were born alive. Two patients had recurrent convulsion i.e. 6.4%. Only one patient had respiratory depression after the use of magnesium sulphate. Conclusion: Eclampsia was well controlled by the use of MgSO/sub 4/. There were only 2 patients who had recurrence of convulsion. Convulsions were controlled in 29 (93.54%) patients despite lack of monitoring facilities of serum magnesium level. (author)

Preparation of Low Molecular Weight Chondroitin Sulfates, Screening of a High Anti-Complement Capacity of Low Molecular Weight Chondroitin Sulfate and Its Biological Activity Studies in Attenuating Osteoarthritis.

Science.gov (United States)

Li, Lian; Li, Yan; Feng, Danyang; Xu, Linghua; Yin, Fengxin; Zang, Hengchang; Liu, Chunhui; Wang, Fengshan

2016-10-11

Chondroitin sulfate (CS) plays important roles in the complement system. However, the CS structure is complicated due to different sources and the number and positions of sulfate groups. The objective of this study was to prepare different low molecular weight chondroitin sulfates (LMWCSs) and to investigate the biological activity in anti-complement capacity. A series of LMWCSs was prepared from different sources and characterized by ultraviolet-visible (UV) spectroscopy, high-performance liquid chromatography (HPLC), size exclusion chromatography-multiangle laser light scattering (SEC-MALLS) and nuclear magnetic resonance (NMR) spectroscopy. Hemolytic, anti-complement 3 deposition capacity and cell viability assays were carried out to investigate the biological activities in vitro. The results showed that LMWCS prepared from shark cartilage with the oxidative degradation method (LMWCS-S-O) had the best anti-complement capacity. LMWCS-S-O could inhibit the alternative pathway of the complement system and protect chondrocytes from cell death. The attenuating effect of LMWCS-S-O on Osteoarthritis (OA) was investigated by destabilization of the medial meniscus (DMM) model in vivo. Functional wind-up, histological and C5b-9 analyses were used to evaluate the treatment effect on the OA model. In vivo results showed that LMWCS-S-O could attenuate OA. LMWCS-S-O with a high content of ΔDi-2,6diS and ΔDi-6S could be used for attenuating OA through regulating the complement system.

Preparation of Low Molecular Weight Chondroitin Sulfates, Screening of a High Anti-Complement Capacity of Low Molecular Weight Chondroitin Sulfate and Its Biological Activity Studies in Attenuating Osteoarthritis

Directory of Open Access Journals (Sweden)

Lian Li

2016-10-01

Full Text Available Chondroitin sulfate (CS plays important roles in the complement system. However, the CS structure is complicated due to different sources and the number and positions of sulfate groups. The objective of this study was to prepare different low molecular weight chondroitin sulfates (LMWCSs and to investigate the biological activity in anti-complement capacity. A series of LMWCSs was prepared from different sources and characterized by ultraviolet-visible (UV spectroscopy, high-performance liquid chromatography (HPLC, size exclusion chromatography-multiangle laser light scattering (SEC-MALLS and nuclear magnetic resonance (NMR spectroscopy. Hemolytic, anti-complement 3 deposition capacity and cell viability assays were carried out to investigate the biological activities in vitro. The results showed that LMWCS prepared from shark cartilage with the oxidative degradation method (LMWCS-S-O had the best anti-complement capacity. LMWCS-S-O could inhibit the alternative pathway of the complement system and protect chondrocytes from cell death. The attenuating effect of LMWCS-S-O on Osteoarthritis (OA was investigated by destabilization of the medial meniscus (DMM model in vivo. Functional wind-up, histological and C5b-9 analyses were used to evaluate the treatment effect on the OA model. In vivo results showed that LMWCS-S-O could attenuate OA. LMWCS-S-O with a high content of ΔDi-2,6diS and ΔDi-6S could be used for attenuating OA through regulating the complement system.

Correlation analysis between sulphate content and leaching of sulphates in recycled aggregates from construction and demolition wastes.

Science.gov (United States)

Barbudo, Auxi; Galvín, Adela P; Agrela, Francisco; Ayuso, Jesús; Jiménez, Jose Ramón

2012-06-01

In some recycled aggregates applications, such as component of new concrete or roads, the total content of soluble sulphates should be measured and controlled. Restrictions are usually motivated by the resistance or stability of the new structure, and in most cases, structural concerns can be remedied by the use of techniques such as sulphur-resistant cements. However, environmental risk assessment from recycling and reuse construction products is often forgotten. The purpose of this study is to analyse the content of soluble sulphate on eleven recycled aggregates and six samples prepared in laboratory by the addition of different gypsum percentages. As points of reference, two natural aggregates were tested. An analysis of the content of the leachable amount of heavy metals regulated by European regulation was included. As a result, the correlation between solubility and leachability data allow suggest a limiting gypsum amount of 4.4% on recycled aggregates. This limit satisfies EU Landfill Directive criteria, which is currently used as reference by public Spanish Government for recycled aggregates in construction works. Copyright © 2012 Elsevier Ltd. All rights reserved.

Modulation of the Plasma Kallikrein-Kinin System Proteins Performed by Heparan Sulfate Proteoglycans

OpenAIRE

Motta, Guacyara; Tersariol, Ivarne L. S.

2017-01-01

Human plasma kallikrein-kinin system proteins are related to inflammation through bradykinin. In the proximity of its target cells, high molecular weight kininogen (H-kininogen) is the substrate of plasma kallikrein, which releases bradykinin from H-kininogen. Heparan sulfate proteoglycans (HSPGs) play a critical role in either recruiting kinin precursors from the plasma, or in the assembly of kallikrein-kinin system components on the cell surface. Furthermore, HSPGs mediate the endocytosis a...

Xyloside effects on in vitro hematopoiesis: Functional and biochemical studies

International Nuclear Information System (INIS)

Kirby, S.L.; Bentley, S.A.

1991-01-01

Xyloside supplementation of long-term bone marrow cultures (LTBMCs) has been reported to result in greatly enhanced proliferation of hematopoietic stem cells. This was presumed to be the result of xyloside-mediated perturbation of proteoglycan synthesis by marrow-derived stromal cells. To investigate this phenomenon, we first studied the effects of xyloside supplementation on proteoglycan synthesis by D2XRadII bone marrow stromal cells, which support hematopoietic stem cell proliferation in vitro. D2XRadII cells were precursor labelled with 35S-sulfate, and proteoglycans separated by ion exchange chromatography, isopyknic CsCl gradient centrifugation, and gel filtration HPLC. Xyloside-supplemented cultures showed an approximately fourfold increase in total 35S incorporation, mainly as free chondroitin-dermatan sulfate (CS/DS) glycosaminoglycan chains in the culture media. Both xyloside supplemented and nonsupplemented cultures synthesized DS1, DS2, and DS3 CS/DS proteoglycans as previously described. In contrast to previous reports, xyloside was found to inhibit hematopoietic cell growth in LTBMC. Inhibitory effects were observed both in cocultures of IL-3-dependent hematopoietic cell lines with supportive stromal cell lines and in primary murine LTBMCs. Xyloside was found to have a marked inhibitory effect on the growth of murine hematopoietic stem cells and IL-3-dependent hematopoietic cell lines in clonal assay systems and in suspension cultures. In contrast, dialyzed concentrated conditioned media from LTBMCs had no such inhibitory effects. These findings suggest that xyloside-mediated inhibition of hematopoietic cell growth in LTBMC resulted from a direct effect of xyloside on proteoglycan synthesis by hematopoietic cells

Effects of glucosamine, chondroitin, or placebo in patients with osteoarthritis of hip or knee: network meta-analysis

DEFF Research Database (Denmark)

Wandel, Simon; Jüni, Peter; Tendal, Britta

2010-01-01

OBJECTIVE: To determine the effect of glucosamine, chondroitin, or the two in combination on joint pain and on radiological progression of disease in osteoarthritis of the hip or knee. Design Network meta-analysis. Direct comparisons within trials were combined with indirect evidence from other t...... and health insurers should not cover the costs of these preparations, and new prescriptions to patients who have not received treatment should be discouraged....... visual analogue scale. DATA SOURCES: Electronic databases and conference proceedings from inception to June 2009, expert contact, relevant websites. Eligibility criteria for selecting studies Large scale randomised controlled trials in more than 200 patients with osteoarthritis of the knee or hip.......02 for interaction). The differences in changes in minimal width of joint space were all minute, with 95% credible intervals overlapping zero. Conclusions Compared with placebo, glucosamine, chondroitin, and their combination do not reduce joint pain or have an impact on narrowing of joint space. Health authorities...

Brittlestars contain highly sulfated chondroitin sulfates/dermatan sulfates that promote fibroblast growth factor 2-induced cell signaling.

Science.gov (United States)

Ramachandra, Rashmi; Namburi, Ramesh B; Ortega-Martinez, Olga; Shi, Xiaofeng; Zaia, Joseph; Dupont, Sam T; Thorndyke, Michael C; Lindahl, Ulf; Spillmann, Dorothe

2014-02-01

Glycosaminoglycans (GAGs) isolated from brittlestars, Echinodermata class Ophiuroidea, were characterized, as part of attempts to understand the evolutionary development of these polysaccharides. A population of chondroitin sulfate/dermatan sulfate (CS/DS) chains with a high overall degree of sulfation and hexuronate epimerization was the major GAG found, whereas heparan sulfate (HS) was below detection level. Enzymatic digestion with different chondroitin lyases revealed exceptionally high proportions of di- and trisulfated CS/DS disaccharides. The latter unit appears much more abundant in one of four individual species of brittlestars, Amphiura filiformis, than reported earlier in other marine invertebrates. The brittlestar CS/DS was further shown to bind to growth factors such as fibroblast growth factor 2 and to promote FGF-stimulated cell signaling in GAG-deficient cell lines in a manner similar to that of heparin. These findings point to a potential biological role for the highly sulfated invertebrate GAGs, similar to those ascribed to HS in vertebrates.

Sulphation of calcium-based sorbents in circulating fluidised beds under oxy-fuel combustion conditions

Energy Technology Data Exchange (ETDEWEB)

Francisco Garcia-Labiano; Luis F. de Diego; Alberto Abad; Pilar Gayan; Margarita de las Obras-Loscertales; Aranzazu Rufas; Juan Adanez [Instituto de Carboquimica (CSIC), Zaragoza (Spain). Dept. Energy and Environment

2009-07-01

Sulphur Retention (SR) by calcium-based sorbents is a process highly dependent on the temperature and CO{sub 2} concentration. In circulating fluidised beds combustors (CFBC's) operating under oxy-fuel conditions, the sulphation process takes place in atmospheres enriched in CO{sub 2} with bed concentrations that can vary from 40 to 95%. Under so high CO{sub 2} concentrations, very different from that in conventional coal combustion atmosphere with air, the calcination and sulphation behaviour of the sorbent must be defined to optimise the SR process in the combustor. The objective of this work was to determine the SO{sub 2} retention capacity of a Spanish limestone at typical oxy-fuel conditions in CFBC's. Long term duration tests of sulphation (up to 24 h), to simulate the residence time of sorbents in CFBC's, were carried out by thermogravimetric analysis (TGA). Clear behaviour differences were found under calcining and non-calcining conditions. Especially relevant was the result obtained at calcining conditions but close to the thermodynamic temperature given for sorbent calcination. This situation must be avoided in CFBC's because the CO{sub 2} produced inside the particle during calcination can destroy the particles if a non-porous sulphate product layer has been formed around the particle. The effect of the main variables on the sorbent sulphation such as SO{sub 2} concentration, temperature, and particle size were analysed in the long term TGA tests. These data were also used to determine the kinetic parameters for the sulphation under oxy-fuel combustion conditions, which were able to adequately predict the sulphation conversion values in a wide range of operating conditions. 20 refs., 5 figs., 2 tabs.

Spore-forming, Desulfosporosinus-like sulphate-reducing bacteria from a shallow aquifer contaminated with gasoline.

Science.gov (United States)

Robertson, W J; Franzmann, P D; Mee, B J

2000-02-01

Previous studies on the geochemistry of a shallow unconfined aquifer contaminated with hydrocarbons suggested that the degradation of some hydrocarbons was linked to bacterial sulphate reduction. There was attenuation of naphthalene, 1,3,5-trimethylbenzene (TMB), toluene, p-xylene and ethylbenzene in the groundwater with concomitant loss of sulphate. Here, the recovery of eight strains of sulphate-reducing bacteria (SRB) from the contaminated site is reported. All were straight or curved rod-shaped cells which formed endospores. Amplification and sequencing of the 16S rDNA indicated that the strains were all sulphate reducers of the Gram-positive line of descent, and were most closely related to Desulfosporosinus (previously Desulfotomaculum) orientis DSM 8344 (97-98.9% sequence similarity). The strains clustered in three phylogenetic groups based on 16S rRNA sequences. Whole cell fatty acid compositions were similar to those of D. orientis DSM 8344, and were consistent with previous studies of fatty acids in soil and groundwater from the site. Microcosms containing groundwater from this aquifer indicated a role for sulphate reduction in the degradation of [ring-UL-14C]toluene, but not for the degradation of [UL-14C]benzene which could also be degraded by the microcosms. Adding one of the strains that was isolated from the groundwater (strain T2) to sulphate-enriched microcosms increased the rate of toluene degradation four- to 10-fold but had no effect on the rate of benzene degradation. The addition of molybdate, an inhibitor of sulphate reduction, to the groundwater samples decreased the rate of toluene mineralization. There was no evidence to support the mineralization of [UL-14C]benzene, [ring-UL-14C]toluene or unlabelled m-xylene, p-xylene, ethylbenzene, TMB or naphthalene by any of the strains in pure culture. Growth of all the strains was completely inhibited by 100 micromol l-1 TMB.

Strategies for neurotrophin-3 and chondroitinase ABC release from freeze-cast chitosan-alginate nerve-guidance scaffolds.

Science.gov (United States)

Francis, Nicola L; Hunger, Philipp M; Donius, Amalie E; Wegst, Ulrike G K; Wheatley, Margaret A

2017-01-01

Freeze casting, or controlled unidirectional solidification, can be used to fabricate chitosan-alginate (C-A) scaffolds with highly aligned porosity that are suitable for use as nerve-guidance channels. To augment the guidance of growth across a spinal cord injury lesion, these scaffolds are now evaluated in vitro to assess their ability to release neurotrophin-3 (NT-3) and chondroitinase ABC (chABC) in a controlled manner. Protein-loaded microcapsules were incorporated into C-A scaffolds prior to freeze casting without affecting the original scaffold architecture. In vitro protein release was not significantly different when comparing protein loaded directly into the scaffolds with release from scaffolds containing incorporated microcapsules. NT-3 was released from the C-A scaffolds for 8 weeks in vitro, while chABC was released for up to 7 weeks. Low total percentages of protein released from the scaffolds over this time period were attributed to limitation of diffusion by the interpenetrating polymer network matrix of the scaffold walls. NT-3 and chABC released from the scaffolds retained bioactivity, as determined by a neurite outgrowth assay, and the promotion of neurite growth across an inhibitory barrier of chondroitin sulphate proteoglycans. This demonstrates the potential of these multifunctional scaffolds for enhancing axonal regeneration through growth-inhibiting glial scars via the sustained release of chABC and NT-3. Copyright © 2014 John Wiley & Sons, Ltd. Copyright © 2014 John Wiley & Sons, Ltd.

Chondroitin sulfate/dermatan sulfate sulfatases from mammals and bacteria.

Science.gov (United States)

Wang, Shumin; Sugahara, Kazuyuki; Li, Fuchuan

2016-12-01

Sulfatases that specifically catalyze the hydrolysis of the sulfate groups on chondroitin sulfate (CS)/dermatan sulfate (DS) poly- and oligosaccharides belong to the formylglycine-dependent family of sulfatases and have been widely found in various mammalian and bacterial organisms. However, only a few types of CS/DS sulfatase have been identified so far. Recently, several novel CS/DS sulfatases have been cloned and characterized. Advanced studies have provided significant insight into the biological function and mechanism of action of CS/DS sulfatases. Moreover, further studies will provide powerful tools for structural and functional studies of CS/DS as well as related applications. This article reviews the recent progress in CS/DS sulfatase research and is expected to initiate further research in this field.

The involvement of proteoglycans in the human plasma prekallikrein interaction with the cell surface.

Directory of Open Access Journals (Sweden)

Camila Lopes Veronez

Full Text Available INTRODUCTION: The aim of this work was to evaluate the role of human plasma prekallikrein assembly and processing in cells and to determine whether proteoglycans, along with high molecular weight kininogen (H-kininogen, influence this interaction. METHODS: We used the endothelial cell line ECV304 and the epithelial cell lines CHO-K1 (wild type and CHO-745 (deficient in proteoglycans. Prekallikrein endocytosis was studied using confocal microscopy, and prekallikrein cleavage/activation was determined by immunoblotting using an antibody directed to the prekallikrein sequence C364TTKTSTR371 and an antibody directed to the entire H-kininogen molecule. RESULTS: At 37°C, prekallikrein endocytosis was assessed in the absence and presence of exogenously applied H-kininogen and found to be 1,418.4±0.010 and 1,070.3±0.001 pixels/cell, respectively, for ECV304 and 1,319.1±0.003 and 631.3±0.001 pixels/cell, respectively, for CHO-K1. No prekallikrein internalization was observed in CHO-745 in either condition. Prekallikrein colocalized with LysoTracker in the absence and presence of exogenous H-kininogen at levels of 76.0% and 88.5%, respectively, for ECV304 and at levels of 40.7% and 57.0%, respectively, for CHO-K1. After assembly on the cell surface, a plasma kallikrein fragment of 53 kDa was predominant in the incubation buffer of all the cell lines studied, indicating specific proteolysis; plasma kallikrein fragments of 48-44 kDa and 34-32 kDa were also detected in the incubation buffer, indicating non-specific cleavage. Bradykinin free H-kininogen internalization was not detected in CHO-K1 or CHO-745 cells at 37°C. CONCLUSION: The prekallikrein interaction with the cell surface is temperature-dependent and independent of exogenously applied H-kininogen, which results in prekallikrein endocytosis promoted by proteoglycans. Prekallikrein proteolysis/activation is influenced by H-kininogen/glycosaminoglycans assembly and controls plasma kallikrein

Cell surface binding and uptake of arginine- and lysine-rich penetratin peptides in absence and presence of proteoglycans

KAUST Repository

Å mand, Helene L.; Rydberg, Hanna A.; Fornander, Louise H.; Lincoln, Per; Nordé n, Bengt; Esbjö rner, Elin K.

2012-01-01

Cell surface proteoglycans (PGs) appear to promote uptake of arginine-rich cell-penetrating peptides (CPPs), but their exact functions are unclear. To address if there is specificity in the interactions of arginines and PGs leading to improved

Human monoclonal IgG selection of Plasmodium falciparum for the expression of placental malaria-specific variant surface antigens

DEFF Research Database (Denmark)

Soerli, J; Barfod, L; Lavstsen, T

2009-01-01

Pregnancy-associated Plasmodium falciparum malaria (PAM) is a major cause of morbidity and mortality in African women and their offspring. PAM is characterized by accumulation of infected erythrocytes (IEs) that adhere to chondroitin sulphate A (CSA) in the placental intervillous space. We show h...... transcription of var2csa. The results corroborate current efforts to develop PAM-specific vaccines based on VAR2CSA....

Studies on calcium, magnesium and sulphate in the Mandovi and Zuari river system (Goa)

Digital Repository Service at National Institute of Oceanography (India)

SenGupta, R.; Naik, S.

-conservative parameters. Sulphate, however behaves in a purely conservative manner and remains in a steady state in the rivers. A'Simple mixture'relationship is applied to calculate the dilution and mixing processes in the rivers using calcium, magnesium and sulphate...

Biotreatment of Cr(VI) contaminated waters by sulphate reducing bacteria fed with ethanol

International Nuclear Information System (INIS)

Pagnanelli, F.; Cruz Viggi, C.; Cibati, A.; Uccelletti, D.; Toro, L.; Palleschi, C.

2012-01-01

Highlights: ► Use of ethanol as electron donor for sulphate-reducing bacteria for the treatment of Cr(VI). ► Isolation of contribution in Cr removal (adsorption vs. bioprecipitation). ► Bioassessment of the process effectiveness by ecotoxicological in vivo tests using C. elegans. - Abstract: Biological treatment of Cr(VI) contaminated waters was performed in fixed bed reactors inoculated with SRB (sulphate-reducing bacteria) growing on ethanol. Treatment efficiency was evaluated by checking chemical abatement of Cr(VI) and by ecotoxicological tests using the nematode Caenorhabditis elegans. A preliminary comparison between ethanol and lactate was performed, denoting that using ethanol, the same values of final sulphate abatement were obtained. In addition ethanol showed to be a substrate more competitive than lactate in kinetic terms. Fixed bed column reactors were continuously fed with a solution containing sulphates (3 g L −1 ), ethanol (1.5 g L −1 ) and Cr(VI) (50 mg L −1 ). At steady state the column inoculated with SRB removed 65 ± 5% of sulphate and 95 ± 5% of chromium. Bioactive removal mechanisms predominated over biosorption. Diminution of Cr(VI) toxicity was assessed by using the nematode C. elegans as a test organism showing that the survival of nematodes was 20% in the presence of the untreated influent and raised up to 53% when the nematodes were exposed to the treated effluent.

Formulation and evaluation and terbutaline sulphate and ...

African Journals Online (AJOL)

We report the use of low rugosity lactose, product of controlled crystallization of this carrier, in the formulation of terbutaline sulphate and beclomethasone dipropionate dry powder inhalers. The deposition patterns obtained with inhalation mixtures consisting of the modified lactose and each of the micronised drugs ...

Magnesium sulphate for women at risk of preterm birth for neuroprotection of the fetus

NARCIS (Netherlands)

Doyle, Lex W.; Crowther, Caroline A.; Middleton, Philippa; Marret, Stephane; Rouse, Dwight

2009-01-01

Background Epidemiological and basic science evidence suggests that magnesium sulphate before birth may be neuroprotective for the fetus. Objectives To assess the effects of magnesium sulphate as a neuroprotective agent when given to women considered at risk of preterm birth. Search strategy We

Purification, structural characterization and antiproliferative properties of chondroitin sulfate/dermatan sulfate from tunisian fish skins.

Science.gov (United States)

Krichen, Fatma; Volpi, Nicola; Sila, Assaâd; Maccari, Francesca; Mantovani, Veronica; Galeotti, Fabio; Ellouz-Chaabouni, Semia; Bougatef, Ali

2017-02-01

Chondroitin sulfate/dermatan sulfate GAGs were extracted and purified from the skins of grey triggerfish (GTSG) and smooth hound (SHSG). The disaccharide composition produced by chondroitinase ABC treatment showed the presence of nonsulfated disaccharide, monosulfated disaccharides ΔDi6S and ΔDi4S, and disulfated disaccharides in different percentages. In particular, the nonsulfated disaccharide ΔDi0S of GTSG and SHSG were 3.5% and 5.5%, respectively, while monosulfated disaccharides ΔDi6S and ΔDi4S were evaluated to be 18.2%, 59% and 14.6%, 47.0%, respectively. Capillary elecrophoresis analysis of GTSG and SHSG contained 99.2% and 95.4% of chondroitin sulfate/dermatan sulfate, respectively. PAGE analysis showed a GTSG and SHSG having molecular masses with average values of 41.72KDa and 23.8KDa, respectively. HCT116 cell proliferation was inhibited (p<0.05) by 70.6% and 72.65% at 200μg/mL of GTSG and SHSG respectively. Both GTSG and SHSG demonstrated promising antiproliferative potential, which may be used as a novel, effective agent. Copyright © 2016 Elsevier B.V. All rights reserved.

Synthesis of Fucosylated Chondroitin Sulfate Glycoclusters: A Robust Route to New Anticoagulant Agents.

Science.gov (United States)

Zhang, Xiao; Yao, Wang; Xu, Xiaojiang; Sun, Huifang; Zhao, Jinhua; Meng, Xiangbao; Wu, Mingyi; Li, Zhongjun

2018-02-01

Fucosylated chondroitin sulfate (FuCS) is a structurally distinct glycosaminoglycan with excellent anticoagulant activity. Studies show that FuCS and its depolymerized fragments exhibit a different anticoagulant mechanism from that of heparin derivatives, with decreased risks of adverse effects and bleeding. However, further exploitation has been hindered by the scarcity of structurally defined oligosaccharides. Herein, facile method is reported for the synthesis of the repeating trisaccharide unit of FuCS based on the degradation of chondroitin sulfate polymers. A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. Further FXase inhibition tests suggested that glycoclusters 33 b-40 b selectively inhibited intrinsic anticoagulant activities, but had little effect on the extrinsic and common coagulation pathways. Notably, glycoclusters with the 2,4-di-O-sulfated fucosyl residue displayed the most potency, which was in consistent with that of natural polysaccharides. These FuCS clusters demonstrated potency to mimic linear glycosaminoglycans and offer a new framework for the development of novel anticoagulant agents. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Chondroitin Sulfate-Rich Extract of Skate Cartilage Attenuates Lipopolysaccharide-Induced Liver Damage in Mice.

Science.gov (United States)

Song, Yeong Ok; Kim, Mijeong; Woo, Minji; Baek, Jang-Mi; Kang, Keon-Hee; Kim, Sang-Ho; Roh, Seong-Soo; Park, Chan Hum; Jeong, Kap-Seop; Noh, Jeong-Sook

2017-06-15

The protective effects of a chondroitin sulfate-rich extract (CSE) from skate cartilage against lipopolysaccharide (LPS)-induced hepatic damage were investigated, and its mechanism of action was compared with that of chondroitin sulfate (CS) from shark cartilage. ICR mice were orally administrated 200 mg/kg body weight (BW) of CS or 400 mg/kg BW of CSE for 3 consecutive days, followed by a one-time intraperitoneal injection of LPS (20 mg/kg BW). The experimental groups were vehicle treatment without LPS injection (NC group), vehicle treatment with LPS injection (LPS group), CS pretreatment with LPS injection (CS group), and CSE pretreatment with LPS injection (CSE group). Hepatic antioxidant enzyme expression levels in the CS and CSE groups were increased relative to those in the LPS group. In LPS-insulted hepatic tissue, inflammatory factors were augmented relative to those in the NC group, but were significantly suppressed by pretreatment with CS or CSE. Moreover, CS and CSE alleviated the LPS-induced apoptotic factors and mitogen-activated protein kinase (MAPK). In addition, CS and CSE effectively decreased the serum lipid concentrations and downregulated hepatic sterol regulatory element-binding proteins expression. In conclusion, the skate CSE could protect against LPS-induced hepatic dyslipidemia, oxidative stress, inflammation, and apoptosis, probably through the regulation of MAPK signaling.

Rapid acquisition of isolate-specific antibodies to chondroitin sulfate A-adherent Plasmodium falciparum isolates in Ghanaian primigravidae

DEFF Research Database (Denmark)

Cox, Sharon E; Staalsoe, Trine; Arthur, Paul

2005-01-01

RBC to chondroitin sulfate A (CSA). The VSA mediating CSA binding (VSA(CSA)) and thus sequestration of pRBC in the placenta are antigenically distinct from those that mediate pRBC sequestration elsewhere in the body, and it has been suggested that VSA(CSA) are relatively conserved and may thus constitute...

Automatic colorimetric determination of low concentrations of sulphate for measuring sulphur dioxide in ambient air

Energy Technology Data Exchange (ETDEWEB)

Persson, G A

1966-01-01

An automatic colorimetric method for the determination of low concentrations of sulphate (0-10 microgram/ml) using the thoron indicator is described. Total amounts of sulphate as small as 0.3 micrograms can be determined. The sulphate is precipitated with barium perchlorate and the excess of barium is indicated with 1-(o-arsenophenylazo)-2-naphthol-3-6-disulfonic acid(thoron). The procedure is worked out primarily for the determination of sulphur dioxide in air after absorption in diluted hydrogen peroxide.

Modelling the competition between sulphate reducers and methanogens in a thermophilic methanol-fed bioreactor

NARCIS (Netherlands)

Spanjers, H.; Weijma, J.; Abusam, A.

2002-01-01

Sulphate can be removed from wastewater by means of biological anaerobic reduction to sulphide. The reduction requires the presence of a substrate that can serve as an electron donor. Methanol a suitable electron donor for sulphate reduction under thermophilic conditions. In an anaerobic system

Performance of Electrocoagulation Process for Removal of Sulphate Ion from Aqueous Environments Using Plate Aluminum Electrodes

Directory of Open Access Journals (Sweden)

a.h Mahvi

2012-05-01

Full Text Available

Background and Objectives: Sulphate ion is one of the main ions available in natural and waste water. The high rate of sulphate in drinking water causes health problems such as irritation and digestive problems and makes water taste bitter. The purpose of this study was to assess the efficiency of sulphate ion removal from drinking water through Electrocogualation process (EC using plate aluminum electrodes. Methods: This study was conducted as a pilot plant using one glass tank in the volume of 1.3liter containing 6 plate aluminum electrodes. These electrodes were attached to a power supply in a mono-polar and parallel arrangement in order to switch the alternating current to the direct one. That is, each electrode was attached to positive and negative poles directly and alternately. The tank was filled with synthetic water containing sulphate ion with the concentration of 350 & 700mg/L. Percentage of sulphate ion removal in potential range of 10, 20 and 30 V, reaction times of 40, 20 and 60min and pH 7.0, 3.0 & 11.0 were measured. Results: In this study the maximum efficiency of sulphate ion removal was in the electrical potential of 30 V, reaction time of 60min and pH 11.0. With increase of ion concentration, the time needed to achieve a suitable efficiency of removal increased, that is, it indicated the direct effect of pH and difference of electrical potential on removal of sulphate ion through EC process. Conclusion: The results of this research show that Electrocogualation technology can be introduced as a suitable and promising technique to remove sulphate ion from aqueous environments using plate aluminum electrodes.

Performance of Electrocoagulation Process for Removal of Sulphate Ion from Aqueous Environments Using Plate Aluminum Electrodes

Directory of Open Access Journals (Sweden)

Rajabizadeh A.

2010-09-01

Full Text Available Background and Objectives: Sulphate ion is one of the main ions available in natural and waste water. The high rate of sulphate in drinking water causes health problems such as irritation and digestive problems and makes water taste bitter. The purpose of this study was to assess the efficiency of sulphate ion removal from drinking water through Electrocogualation process (EC using plate aluminum electrodes.Methods: This study was conducted as a pilot plant using one glass tank in the volume of 1.3liter containing 6 plate aluminum electrodes. These electrodes were attached to a power supply in a mono-polar and parallel arrangement in order to switch the alternating current to the direct one. That is, each electrode was attached to positive and negative poles directly and alternately. The tank was filled with synthetic water containing sulphate ion with the concentration of 350 & 700mg/L. Percentage of sulphate ion removal in potential range of 10, 20 and 30 V, reaction times of 40, 20 and 60min and pH 7.0, 3.0 & 11.0 were measured.Results: In this study the maximum efficiency of sulphate ion removal was in the electrical potential of 30 V, reaction time of 60min and pH 11.0. With increase of ion concentration, the time needed to achieve a suitable efficiency of removal increased, that is, it indicated the direct effect of pH and difference of electrical potential on removal of sulphate ion through EC process.Conclusion: The results of this research show that Electrocogualation technology can be introduced as a suitable and promising technique to remove sulphate ion from aqueous environments using plate aluminum electrodes.

Sulphate Potasium Extraction From Banana Stem Ash with Bleaching Earth Waste Liquid

OpenAIRE

Edahwati, Luluk

2010-01-01

Sulphate potassium is one of the important chemistry compound for industrial at our country, usually for fertilizerindustry. Therefore, necessary done sulphate potassium maker watchfulness from various ingredient that containsulphate potassium compound. Among others with extract banana stem ash. Watchfulness methodology that is withmix banana stem ash with aquadest (20 gram ash/1 water litre) in temperature 100oc during 30 minutes and stirringrotation 300 rpm. Ash extract that got reaction wi...

Chondroitinase C Selectively Degrades Chondroitin Sulfate Glycosaminoglycans that Inhibit Axonal Growth within the Endoneurium of Peripheral Nerve.

Directory of Open Access Journals (Sweden)

James B Graham

Full Text Available The success of peripheral nerve regeneration is highly dependent on the regrowth of axons within the endoneurial basal lamina tubes that promote target-oriented pathfinding and appropriate reinnervation. Restoration of nerve continuity at this structural level after nerve transection injury by direct repair and nerve grafting remains a major surgical challenge. Recently, biological approaches that alter the balance of growth inhibitors and promoters in nerve have shown promise to improve appropriate axonal regeneration and recovery of peripheral nerve function. Chondroitin sulfate proteoglycans (CSPGs are known inhibitors of axonal growth. This growth inhibition is mainly associated with a CSPG's glycosaminoglycan chains. Enzymatic degradation of these chains with chondroitinase eliminates this inhibitory activity and, when applied in vivo, can improve the outcome of nerve repair. To date, these encouraging findings were obtained with chondroitinase ABC (a pan-specific chondroitinase. The aim of this study was to examine the distribution of CSPG subtypes in rodent, rabbit, and human peripheral nerve and to test more selective biological enzymatic approaches to improve appropriate axonal growth within the endoneurium and minimize aberrant growth. Here we provide evidence that the endoneurium, but not the surrounding epineurium, is rich in CSPGs that have glycosaminoglycan chains readily degraded by chondroitinase C. Biochemical studies indicate that chondroitinase C has degradation specificity for 6-sulfated glycosaminoglycans found in peripheral nerve. We found that chondroitinase C degrades and inactivates inhibitory CSPGs within the endoneurium but not so much in the surrounding nerve compartments. Cryoculture bioassays (neurons grown on tissue sections show that chondroitinase C selectively and significantly enhanced neuritic growth associated with the endoneurial basal laminae without changing growth-inhibiting properties of the surrounding

Toxicology studies of primycin-sulphate using a three-dimensional (3D) in vitro human liver aggregate model.

Science.gov (United States)

Pénzes, Ágota; Mahmud Abdelwahab, Elhusseiny Mohamed; Rapp, Judit; Péteri, Zsanett A; Bovári-Biri, Judit; Fekete, Csaba; Miskei, György; Kvell, Krisztián; Pongrácz, Judit E

2017-11-05

Primycin-sulphate is a highly effective compound against Gram (G) positive bacteria. It has a potentially synergistic effect with vancomycin and statins which makes primycin-sulphate a potentially very effective preparation. Primycin-sulphate is currently used exclusively in topical preparations. In vitro animal hepatocyte and neuromuscular junction studies (in mice, rats, snakes, frogs) as well as in in vitro human red blood cell experiments were used to test toxicity. During these studies, the use of primycin-sulphate resulted in reduced cellular membrane integrity and modified ion channel activity. Additionally, parenteral administration of primycin-sulphate to mice, dogs, cats, rabbits and guinea pigs indicated high level of acute toxicity. The objective of this study was to reveal the cytotoxic and gene expression modifying effects of primycin-sulphate in a human system using an in vitro, three dimensional (3D) human hepatic model system. Within the 3D model, primycin-sulphate presented no acute cytotoxicity at concentrations 1μg/ml and below. However, even at low concentrations, primycin-sulphate affected gene expressions by up-regulating inflammatory cytokines (e.g., IL6), chemokines (e.g., CXCL5) and by down-regulating molecules of the lipid metabolism (e.g., peroxisome proliferator receptor (PPAR) alpha, gamma, etc). Down-regulation of PPAR alpha cannot just disrupt lipid production but can also affect cytochrome P450 metabolic enzyme (CYP) 3A4 expression, highlighting the need for extensive drug-drug interaction (DDI) studies before human oral or parenteral preparations can be developed. Copyright © 2017 Elsevier B.V. All rights reserved.

Effects of Manganese (Ii Sulphate on Structural, Spectral, Optical, Thermal and Mechanical Properties of L-Alanine Sodium Sulphate Single Crystals

Directory of Open Access Journals (Sweden)

F. Praveena

2017-04-01

Full Text Available New Non-linear Optical materials have been attracting in the research world for their potential applications in emerging opto-electronic technology. The dipolar nature of amino acid leads to peculiar physical and chemical properties, thus making a good candidate for NLO applications. Single crystals of manganese(II sulphate doped L-Alanine sodium sulphate(LASS has been synthesized by slow evaporation technique. Structural property of the grown crystals are characterized by X-ray powder diffraction,FT-IR spectral analysis conforms all the functional groups. Thermogravity (TG and differential themogravimetric (DTA analysis have been performed to study the thermal stability of the crystals. The second harmonic generation efficiency was measured by Kurtz-Perry powder technique. The transmission and absorption of electromagnetic radiation is analysed through UV-VIS spectrum. Microhardness was measured at different applied load to understand the mechanical stability of the crystal.

Evaluating portland cement concrete degradation by sulphate exposure through artificial neural networks modeling

International Nuclear Information System (INIS)

Oliveira, Douglas Nunes de; Bourguignon, Lucas Gabriel Garcia; Tolentino, Evandro; Costa, Rodrigo Moyses; Tello, Cledola Cassia Oliveira de

2015-01-01

A concrete is durable if it has accomplished the desired service life in the environment in which it is exposed. The durability of concrete materials can be limited as a result of adverse performance of its cement-paste matrix or aggregate constituents under either chemical or physical attack. Among other aggressive chemical exposures, the sulphate attack is an important concern. Water, soils and gases, which contain sulphate, represent a potential threat to the durability of concrete structures. Sulphate attack in concrete leads to the conversion of the hydration products of cement to ettringite, gypsum, and other phases, and also it leads to the destabilization of the primary strength generating calcium silicate hydrate (C-S-H) gel. The formation of ettringite and gypsum is common in cementitious systems exposed to most types of sulphate solutions. The present work presents the application of the neural networks for estimating deterioration of various concrete mixtures due to exposure to sulphate solutions. A neural networks model was constructed, trained and tested using the available database. In general, artificial neural networks could be successfully used in function approximation problems in order to approach the data generation function. Once data generation function is known, artificial neural network structure is tested using data not presented to the network during training. This paper is intent to provide the technical requirements related to the production of a durable concrete to be used in the structures of the Brazilian near-surface repository of radioactive wastes. (author)

Biotreatment of Cr(VI) contaminated waters by sulphate reducing bacteria fed with ethanol

Energy Technology Data Exchange (ETDEWEB)

Pagnanelli, F., E-mail: francesca.pagnanelli@uniroma1.it [Department of Chemistry, Sapienza University of Rome, P.le Aldo Moro 5, 00185 Rome (Italy); Cruz Viggi, C., E-mail: carolina.cruzviggi@uniroma1.it [Department of Chemistry, Sapienza University of Rome, P.le Aldo Moro 5, 00185 Rome (Italy); Cibati, A., E-mail: alessio.cibati@uniroma1.it [Department of Chemistry, Sapienza University of Rome, P.le Aldo Moro 5, 00185 Rome (Italy); Uccelletti, D., E-mail: daniela.uccelletti@uniroma1.it [Department of Biology and Biotechnology, Sapienza University of Rome, P.le A. Moro 5, 00185 Rome (Italy); Toro, L., E-mail: luigi.toro@uniroma1.it [Department of Chemistry, Sapienza University of Rome, P.le Aldo Moro 5, 00185 Rome (Italy); Palleschi, C., E-mail: claudio.palleschi@uniroma1.it [Department of Biology and Biotechnology, Sapienza University of Rome, P.le A. Moro 5, 00185 Rome (Italy)

2012-01-15

Highlights: Black-Right-Pointing-Pointer Use of ethanol as electron donor for sulphate-reducing bacteria for the treatment of Cr(VI). Black-Right-Pointing-Pointer Isolation of contribution in Cr removal (adsorption vs. bioprecipitation). Black-Right-Pointing-Pointer Bioassessment of the process effectiveness by ecotoxicological in vivo tests using C. elegans. - Abstract: Biological treatment of Cr(VI) contaminated waters was performed in fixed bed reactors inoculated with SRB (sulphate-reducing bacteria) growing on ethanol. Treatment efficiency was evaluated by checking chemical abatement of Cr(VI) and by ecotoxicological tests using the nematode Caenorhabditis elegans. A preliminary comparison between ethanol and lactate was performed, denoting that using ethanol, the same values of final sulphate abatement were obtained. In addition ethanol showed to be a substrate more competitive than lactate in kinetic terms. Fixed bed column reactors were continuously fed with a solution containing sulphates (3 g L{sup -1}), ethanol (1.5 g L{sup -1}) and Cr(VI) (50 mg L{sup -1}). At steady state the column inoculated with SRB removed 65 {+-} 5% of sulphate and 95 {+-} 5% of chromium. Bioactive removal mechanisms predominated over biosorption. Diminution of Cr(VI) toxicity was assessed by using the nematode C. elegans as a test organism showing that the survival of nematodes was 20% in the presence of the untreated influent and raised up to 53% when the nematodes were exposed to the treated effluent.

Evaluating portland cement concrete degradation by sulphate exposure through artificial neural networks modeling

Energy Technology Data Exchange (ETDEWEB)

Oliveira, Douglas Nunes de; Bourguignon, Lucas Gabriel Garcia; Tolentino, Evandro, E-mail: tolentino@timoteo.cefetmg.br [Centro Federal de Educacao Tecnologica de Minas Gerais (CEFET-MG), Timoteo, MG (Brazil); Costa, Rodrigo Moyses, E-mail: rodrigo@moyses.com.br [Universidade de Itauna, Itauna, MG (Brazil); Tello, Cledola Cassia Oliveira de, E-mail: tellocc@cdtn.br [Centro de Desenvolvimento da Tecnologia Nucelar (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)

2015-07-01

A concrete is durable if it has accomplished the desired service life in the environment in which it is exposed. The durability of concrete materials can be limited as a result of adverse performance of its cement-paste matrix or aggregate constituents under either chemical or physical attack. Among other aggressive chemical exposures, the sulphate attack is an important concern. Water, soils and gases, which contain sulphate, represent a potential threat to the durability of concrete structures. Sulphate attack in concrete leads to the conversion of the hydration products of cement to ettringite, gypsum, and other phases, and also it leads to the destabilization of the primary strength generating calcium silicate hydrate (C-S-H) gel. The formation of ettringite and gypsum is common in cementitious systems exposed to most types of sulphate solutions. The present work presents the application of the neural networks for estimating deterioration of various concrete mixtures due to exposure to sulphate solutions. A neural networks model was constructed, trained and tested using the available database. In general, artificial neural networks could be successfully used in function approximation problems in order to approach the data generation function. Once data generation function is known, artificial neural network structure is tested using data not presented to the network during training. This paper is intent to provide the technical requirements related to the production of a durable concrete to be used in the structures of the Brazilian near-surface repository of radioactive wastes. (author)

Hexagonal ZnO porous plates prepared from microwave synthesized layered zinc hydroxide sulphate via thermal decomposition

Energy Technology Data Exchange (ETDEWEB)

Machovsky, Michal, E-mail: machovsky@ft.utb.cz [Centre of Polymer Systems, University Institute, Tomas Bata University in Zlin, Nad Ovcirnou 3685, 760 01 Zlin (Czech Republic); Polymer Centre, Faculty of Technology, Tomas Bata University in Zlin, Nam. T.G. Masaryka 275, 762 72 Zlin (Czech Republic); Kuritka, Ivo, E-mail: ivo@kuritka.net [Centre of Polymer Systems, University Institute, Tomas Bata University in Zlin, Nad Ovcirnou 3685, 760 01 Zlin (Czech Republic); Polymer Centre, Faculty of Technology, Tomas Bata University in Zlin, Nam. T.G. Masaryka 275, 762 72 Zlin (Czech Republic); Sedlak, Jakub, E-mail: j1sedlak@ft.utb.cz [Centre of Polymer Systems, University Institute, Tomas Bata University in Zlin, Nad Ovcirnou 3685, 760 01 Zlin (Czech Republic); Polymer Centre, Faculty of Technology, Tomas Bata University in Zlin, Nam. T.G. Masaryka 275, 762 72 Zlin (Czech Republic); Pastorek, Miroslav, E-mail: pastorek@ft.utb.cz [Centre of Polymer Systems, University Institute, Tomas Bata University in Zlin, Nad Ovcirnou 3685, 760 01 Zlin (Czech Republic); Department of Polymer Engineering, Faculty of Technology, Tomas Bata University in Zlin, Nam. T.G. Masaryka 275, 762 72 Zlin (Czech Republic)

2013-10-15

Graphical abstract: - Highlights: • Zinc hydroxy sulphate was synthesized in 3 min via microwave hydrothermal route. • Zinc hydroxy sulphate was converted into mesh like porous ZnO by calcining at 900°. • The process of transformation is topotactic. - Abstract: Layered zinc hydroxide sulphate (ZHS) was prepared by microwave-assisted hydrothermal precipitation of zinc sulphate monohydrate with hexamethylenetetramine. Under ambient conditions, the structure of ZHS determined by X-ray diffraction (XRD) was found to be a mixture of zinc hydroxide sulphate pentahydrate Zn{sub 4}SO{sub 4}(OH){sub 6}·5H{sub 2}O and tetrahydrate Zn{sub 4}SO{sub 4}(OH){sub 6}·4H{sub 2}O. Fourier transform infrared (FTIR) spectroscopy was used for characterization of the prepared materials. Based on the interpretation of ZHS's thermal decomposition profile obtained by thermogravimetric analysis, ZnO of high purity was prepared by calcination at 900 °C for 2 h. The structure of the resulting ZnO was confirmed by the XRD. The morphology examination by scanning electron microscopy revealed a porous mesh-like ZnO structure developed from the ZHS precursor at the expense of mass removal due to the release of water and sulphate during the calcination.