A full assignment of proton resonances for an alpha(1-3)-linked fucose residue in keratan sulphate from bovine articular cartilage

Full proton NMR assignments have been achieved for the alpha(1-3)-linked fucose residues contained in alkaline borohydride reduced keratan sulphate chains derived from bovine articular cartilage. This involved 500 MHz spectroscopy at 60 degrees C and included COSY and RELAYED-COSY determinations

Structural and immunological studies of keratan sulphates from mature bovine articular cartilage.

Two populations of alkaline-borohydride-reduced keratan sulphate (KS) chains were prepared from the two peptido-keratan sulphate trypsin fragments of proteoglycan aggregates isolated from bovine femoral head cartilage (6-year-old animals). Each population was separated by high-performance ion-exchange chromatography on a Pharmacia Mono-Q column into eight pools, Q1-Q8. These were analysed by gel permeation chromatography, radioimmunoassay with the monoclonal antibody MZ15, and 500 MHz 1H n.m.r. spectroscopy. Upon chromatography on Sephadex G-75 the Mono-Q fractions were shown to increase in hydrodynamic size progressively from Q1 to Q8 for both KS populations. For each population the strongest antigenic response with the anti-KS monoclonal antibody MZ15 was expressed by the two fractions of greatest size and charge density, Q7 and Q8. Proton n.m.r. spectroscopic studies on the two series of fractions demonstrated: (i) a progressive increase in the level of galactose sulphation from Q1 to Q8, (ii) the presence of approximately one alpha(1-3)-linked fucose residue per chain in every sample, and (iii) the presence of N-acetylneuraminic acids in three discrete environments, two alpha(2-3)- and one alpha(2-6)-linked in every sample. The results are discussed in terms of a possible heterogeneity in the carbohydrate-protein linkage region of keratan sulphates from bovine articular cartilage

Aspectos da distribuição de tecidos músculo-esqueléticos de um banco de tecidos Aspects of the distribution of musculoskeletal tissues by a tissue bank

OBJETIVO: Avaliar as características da distribuição desses por um Banco de Tecidos no Brasil. MÉTODOS: Base de dados do Banco de Tecidos entre setembro de 2006 e junho de 2008. Características dos receptores foram tabuladas. Os tipos de tecidos processados foram: cabeças femorais, osso metafisio-epifisário, osso cortical, ossos curtos ou chatos e tendões. O destino dos enxertos foi analisado. Frequências das distribuições foram obtidas e analisadas. RESULTADOS: Foram distribuídas 734 unidades tecidos fresco-congelados, transplantadas em 683 receptores. Doadores de múltiplos órgãos originaram 97,9% dos tecidos e doadores vivos os demais. Foram transplantados 489 unidades de osso córtico-esponjoso, 137 de osso metafisio-epifisário, 44 de osso chato ou curto, 3 de tendão, 29 de osso particulado e 32 de cabeças femorais. A média de idade dos receptores foi 50,3 anos, sendo 59,5% do sexo feminino e 40,5% do masculino. Os tecidos foram destinados para uso ortopédico em 21,1% dos casos e buco-maxilo-facial, em 78,9%. CONCLUSÃO: O Banco de Tecidos aumentou o número de distribuições em resposta à demanda crescente de tecidos, principalmente para uso em cirurgia buco-maxilo-facial.<br>OBJECTIVE: Is to evaluate the characteristics of the distribution of these grafts by a Tissue Bank in Brazil. METHODS: Tissue Bank database from September 2006 to June 2008. The characteristics of the recipients were drawn up in the table form. The types of tissue processed were: femoral heads, metaphyseal-epiphyseal bone, cortical bone, flat or short bones and tendons. The intended purpose of the grafts was analyzed, and distribution frequencies were also obtained and analyzed. RESULTS: Altogether, 734 units of fresh-frozen tissue were distributed and transplanted into 683 recipients. In terms of origin of the tissues, 97.9% came from multiple organ donors, and the remainder from living donors. A total of 489 units of cortical bone were transplanted, 137 of metaphyseal-epiphyseal bone, 44 of short or flat bones, 3 of tendon, 29 of particulate bone and 32 femoral heads. The mean age of the recipients was 50.3 years; 59.5% were women and 40.5% men. The tissues were used in orthopedic surgeries in 21.1% of the cases, and in oral and maxillofacial procedures in 78.9%. CONCLUSION: The Tissue Bank has increased the number of distributions in response to the growing demand for tissues, particularly for use in oral and maxillofacial procedures

Oligosaccharides Derived by Keratanase II Digestion of Bovine Articular Cartilage Keratan Sulphates.

Alkaline borohydride-reduced keratan sulphate chains from bovine articular cartilage (6–8-year-old animals) were subjected to a limit digest with the enzyme keratanase II. Using 1H-NMR spectroscopy, 25 reduced oligosaccharides deriving from keratan sulphate were shown to have the following structures [GlcNAc(6S)-ol represents N -acetylglucosaminitol 6-O -sulphate]: Galβ1–4-GlcNAc(6S)-ol, Galβ1-4GlcNAc(6S) β1–3Galβ1–4GlcNAc(6S)-ol, Gal(6S) β1–4GlcNAc(6S)-ol, Gal-(6S) β1–4GlcNAc(6S) β-1–3 Galβ-4GlcNAc(6S)-ol, Galβ1–4GlcNAc(6S) β1–3Gal(6S) β1–4GlcNAc(6S)-ol, Gal(6S) β1–4GlcNAc(6S) β1–3Gal(6S)1–4GlcNAc(6S)-ol, Galβ1–4(Fuca1–3)GlcNAc(6S)-ol, Galβ–4-(Fucα1–3)GlcNAc(6S) β1–3Galβ1–4(Fcα1–3)G1cNAc(6S)-ol, Galβ1–4GlcNAc(6S) β1–3Galβ1–4(Fucα1–3)-GlcNAc(6S)-ol, Galβ1–3(Fucα1–3)GlcNAc(6S) β1–3Galβ1–4GlcNAc(6S)-ol, Gal(6S) β1–4GlcNAc-(6S) β1–3Galβ1–4(Fucα1–3)GlcNAc(6S)-ol, Galβ1–4(Fucα1–3)GlcNAc(6S) β1–3Gal(6S) β1–4GlcNAc(6S)-ol, Galβ1–4 GlcNAc(6S) β1–6(Galβ1–3)GalNAc-ol, Galβ1–4 GlcNAc(6S) β1–6(NeuAc2–3Galβ1–3)Gal-Nac-ol, Galβ1–4GclNAc(6S) β1–3Galβ1–4GlcNAc(6S) β1–4GlcNAc(6S) β1–6(Galβ1–3)GalNAc-ol, Gal(6S) β1–4GlcNAc-(6S) β1–6(Galβ1–3)GalNAc-ol. Galβ1–4GlcNAc(6S) β1–3Galβ1–4GlcNAc (6S) β1–6(NeuAc2–3Galβ1–3)-GalNAc-ol. Gal(6S) β1–4GlcNAc-(6S) β1–6(NeuAcα2–3Galβ1–3)GalNAc-ol. Gal(6S) β1–4GlcNAc-(6S) β1–3Galβ1–4GlcNAc(6S) β1–6(Galβ1–3)GalNAc-ol, Gal(6S) β1–4GlcNAc(6S)β1–3Galβ1–4GlcNAc- (6S) β1–6(NeuAcα2–3Galβ1–3)GalNAc-ol,NeuAcα2–6Galβ1–4 GalNAc(6S) β1–3Galβ1–4 GlcNAc(6s)-ol, NecAcα2–3Galβ1–4GlcNAC(6s)β1–3Galβ1–4GlcNAc(6S)-ol, NeuAcα2–6Galβ1–4GlcNAc(6S)β1–3Gal-(6S)β1–4GlcNAc(6S)-ol, NeuAcα2–3Galβ1–4GlcNAc(6S) β1–3Gal(6S)β1–4GlcNAc(6S)-ol and Neu-Acα2–3Gal(6S) β1–4GlcNAc(6S)β1–3Gal(6Sβ)1–4GlcNAc(6S)-ol. Proton chemical shifts for these oligosaccharides were assigned using one- and two-dimensional NMR spectroscopic methods. These results confirm the findings of Nakazawa et al. [Nakazawa, K., Ito, M., Yamagata, T. and Suzuki, S. (1989) in Keratan sulphate: chemistry, biology and chemical pathology (Greiling, H. and Scott, J. E., eds) pp. 99–110, The Biochemical Society, London], namely that keratanase II cleaves the O -glycosidic bond of a β (1–3)-linked 6-O -sulphated N -acetylglucosamine. However, the internal sulphated N -acetylglucosamine in the sialylated capping oligosaccharides is not cleaved because of the proximity of the sialic acid residue. In addition, keratanase II is the only degradative method examined so far which can cleave the glycosidic bond of a fucosylated N -acetylglucosamine residue as fucose residues confer resistance to both keratanase and hydrazino-lysisnitrous acid fragmentation