IR spectroscopy of mucopolysaccharides in aqueous solutions

Ovsepyan, A. M.; Kobyakov, V. V.; Панов, В. П.

doi:10.1007/bf00615614

Cited by 4 publications

(2 citation statements)

References 8 publications

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“…Despite these advantages, and although there have been numerous studies of extracellular sulphated polysaccharides by vibrational spectroscopy (e.g. Orr, 1954;Suzuki & Strominger, 1960;Cael et al, 1973Cael et al, , 1976Ovsepyan et al, 1977Ovsepyan et al, , 1979Cabassi et al, 1978;Casu et al, 1978;Bychkov et al, 1981;Panov & Ovsepyan, 1984;Greer & Yaphe, 1984;Rochas et al, 1986;Zablackis & Santos, 1986), the complex and variable nature of the spectra produced has particularly limited examination of the complex heparin and heparan sulphate glycosaminoglycans by this technique. Although interpretation of spectra is facilitated by careful definition of polymer environment, and by use of glycosaminoglycan preparations that have been characterized chemically and enzymically (Grant et al, 1987a(Grant et al, , 1989), technical problems can still make spectral interpretation awkward: light-scattering by the polymer may complicate spectra in some cases, and the use of an inert dispersant of suitable refractive index such as Nujol to avoid this causes superimposition of dispersant and polymer absorbances.…”

Section: Introductionmentioning

confidence: 99%

Infrared spectroscopy of heparins suggests that the region 750-950 cm-1 is sensitive to changes in iduronate residue ring conformation

Grant

Long

Moffat

et al. 1991

Biochemical Journal

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By careful definition of polymer environment, heparin i.r. spectra were examined in a region (750-950 cm-1) in which sulphate half-ester absorptions occur. Changes seen in this region when metal ion-heparin complexes are converted into heparinic acid, when heparin is carboxy-group-reduced and when various concentrations of Li(+)-heparin are examined are tentatively interpreted in terms of changes in the ring conformation of iduronate residues.

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Section: Introductionmentioning

confidence: 99%

Infrared spectroscopy of heparins suggests that the region 750-950 cm-1 is sensitive to changes in iduronate residue ring conformation

Grant

Long

Moffat

et al. 1991

Biochemical Journal

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“…1 0 I s), enables small quantities of sample obtained from defined cell sources to be studied, allows the study of solid, gel and dissolved samples, and permits the influence of such factors as counter-cation type and the molecular structure of water molecules associated with the polymer to be evaluated [8]. Despite these advantages, and although there have been numerous studies of extracellular polysaccharides by vibrational spectroscopy [9][10][11][12][13][14][15][16][17][18][19][20][21][22][23], the complex and variable nature of the spectra produced has limited examination of the more complex glycosaminoglycan polysaccharides by this technique. Certainly, i.r.…”

mentioning

confidence: 99%

Infrared spectroscopy as a method for investigating the conformations of iduronate saccharide residues in glycosaminoglycans

Grant

Long

Moffat

et al. 1990

Biochemical Society Transactions

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Newc.asrle-icpon-Tyne, NE2 4HH, U.K. and ?Ear, Nose and Throat Department, Freeman Hospital, Newcastle-upon-Tyrie NE2 411t1, U.K.media with effusion is characterized by the accumulation of a sterile viscous fluid in the middle ear cleft which is not cleared by the mucociliary transport system [ 11. Previous studies have shown that high molecular mass mucins, with a polymeric structure based on disulphide bridges, comprise 18-31% of the non-dialysable solids in thick effusions [2] and that these mucins are a major viscous component of the effusion 131. Middle ear mucins have a composition characteristic of other mucins and are rich in glucosamine (17.1% of freeze-dried weight) [ 2 ] . At present nothing is known about the biosynthesis or rates of secretion of mucins in normal subjects and patients with otitis media with effusion.Here we describe an in virro system for the study of the biosynthesis and secretion of middle ear mucins and the control mechanisms thereof.Explants of middle ear mucosa (approx. 0.5-1.0 mm') were removed from the tympanum of consenting patients undergoing middle ear surgery. Explants were supported by nucleopore filters floated on medium [RPMI 1640 containing 10%, (v/v) fetal calf serum, 2 mwglutamate, 1% (v/v) penicillin/streptomycin and 2.5 m~-fungizone] and incubated at 37°C in 5% CO,. After 48 h incubation, [ I4Cc]glucosamine was added to the incubation medium ( 2 pCi/ml final concentration) and the medium was collected after a further 24 h. Tissue viability was assessed histologically at the end of the incubation after haemotoxylin and eosin, and periodic acid-Schiff staining. Free label was removed from the medium by exhaustive dialysis. The labelled macromolecules ( M , > 1 0 000) recovered were characterized in terms of their hydrodynamic size, buoyant density and susceptibility t o various enzymes. Hydrodynamic size, before and after enzymic digestion was analysed by size exclusion chromatography on a Sephadex G-150 column (80cm x 1.5 cm) eluted with 0.2 ~-NaC1/0.2% (w/v) NaN,. The buoyant density of radiolabelled peaks was measured after centrifugation (48 h, 40000 rev./min at 4°C) in CsCl density gradients (starting density I .42 g/ml) 14). Labelled macromolecules were digested with papain (EC 3.4.22.1; 0.08 mg papain/ml of medium in 67 mM-sodium phosphate buffer, pH 6.7, containing 5 mM-cysteine/HCI and 5 mM-EDTA, for 48 h at 60°C [SI), chondroitinase ABC (EC 4.2.2.4; 3.2 pg chondroitinase/ml of medium in 0.1 M-sodium acetate/(). 1 M-Tris, pH 7.3, for 4 h at 37°C [6]) and heparinase Ill (EC 4.2.2.8; in 0.01 M-sodium phosphate, pH 7.0, for 16 h at 25°C 171). The activity o f the enzymes was confirmed by parallel digestions of specific substrates.Histology showed that explants could be maintained in culture for up to 7 days. Previous studies have reported the maintenance of human middle ear mucosa in organ culture and measurement of ciliary activity [ 81; however, biosynthesis and secretion of macromolecules were not measured in that study.After gel filtration of labelled macrom...

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Molecular and functional characteristics of purified gum from Australian chia seeds

Timilsena

Adhikari

Kasapis

et al. 2016

Carbohydrate Polymers

192

119

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IR spectroscopy of mucopolysaccharides in aqueous solutions

Cited by 4 publications

References 8 publications

Infrared spectroscopy of heparins suggests that the region 750-950 cm-1 is sensitive to changes in iduronate residue ring conformation

Infrared spectroscopy of heparins suggests that the region 750-950 cm-1 is sensitive to changes in iduronate residue ring conformation

Infrared spectroscopy as a method for investigating the conformations of iduronate saccharide residues in glycosaminoglycans

Molecular and functional characteristics of purified gum from Australian chia seeds

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